-------
TABLE 2. (CON'T)
1977 PRODUCTIVITY, CHLOROPHYLL A, AND ASSIMILATION RATIO A 
JAN
FEB
A8
A9
A10
All
A12
PROD
CHL A
ASMA
PROD
CHLA
ASMA
-
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
1 .00
1 .03
0.97
1.31
1 .14
1.15
"I
1 .29
0.30
4 .30
0.86
0.85
1.01
0.67
0 .72
0.93
MAR APR
	1	
MAY
JUNE
JULY AUG
SEPT
OCT
NOV
DEC
2.98 j 13.91
1.56j 6.56
1.911 2.12
I
I
I
I
I
I
3.05 j
1.34 |
2 . 2 8 J
I
I
I
6.04
2 .36
2 .52
3.42)16.18
1.34] 4.26
2.551 3.80
3.82J 19.41
1.49]
2.561
I
6.88]
2.8a
1 .72]
1 .471
1. 17j
I
I
	,
* * * *i
4 .701
*~**!
10.37
2.49
4. 16
10 .09
5 .78
1 .75
7.99
4.00
2.00
* * ~ *
2.62
* * * *
4 .20
2.13
1 .97
4
34
83
23
4 . 16
0.82
5.07
3.39
1 .55
2.19
8.59
~ * * *
2.84
* * * *
****
15.89
2 . 33
6.82
9.56
1 .67
5.72
12.95
2.42
5.35
14 .38
2.20
6.54
11.17
2.22
5.03
40.76
3.64
11 . 20
18.38
3.50
5.25
13.80
4.10
3.37
11.02
3.38
3.26
9.74
4.82
2.02
43.03
3.90
11.03
39 . 1 1
1 . 76
22.22
35.73
3.01
11.87
I	
39 .83
2.67
14 .92
12.47
2.09
5.97
0.82
2.66
0.31
T	T	
3.71 '4.00
**** 13.72
**** \l
I
0 ,,45
2.31
0.19
0.20
3.06
0.07
1	.00
2	.63
0.38
0.28
2. 14
0.13
08
3.77*J
2 .29*|
1 .65*'
4.23 |3.65
4.47 j 2.76
0.95 j 1.32
3.7 7 * J
4.60*[
0.82*1
+

I ~ * * *
J ****
I ****
I
.1	I
****
* * * *
* + **
J * + **
I ****
I * * * *
I
2.01
2 .43
0.83
j 1.43
J 0.76
I 1 .88
1 .78*]
1 .72 j
1 .03=1
	L
0.40
0.91
0.44
0.62
0.98
0.63
* * * *
*#* *
* * * *
~ * * *
****
****
1 .67
2.62
0.64
~SPECIAL REPORT DATED OCTOBER 12, 1977
VALUES OF **** REPRESENT DATA NOT AVAILABLE
4*

-------
A 1 3
A14
A1 5
A 1 6
PROD
CHL A
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
TABLE 2 . (CON'T)
1977 PRODUCTIVITY, CHLOROPHYLL A, AND ASSIMILATION RATIO A (CON'T)
JAN
FEB
MAR
APR
MAY
JUNE
JULY AUG
SEPT
OCT
NOV
DEC
PROD
CHLA
ASMA
* * * *
* * * *
~ * * *
* * * *
* # * *
* * * *
* * * *
* + * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * « *
* * * *
* * * *
* ~ * *
* * * *
~ * * *
* * * *
* * * *
* * # *
****
* * * *
* * * *
* * * *
* * # *
~ * * *
* * * *
****
****
* * * -*
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * *
* * * #
* * + *
* * * #
* * * *
* * ~ *
1
1
1
1
** * ~
1 1
i 1
1 *** * 1
| |
i
i
* * * * i
i
* * * *
i i
i i
j 10.67 j
1
I
1 . 93 J
2.15
* * * +
1
* * * *
1 I
**** i
* * * *
1 5.26 1
0.621
1.18
* ~ * *
1
1
* * * *
{ * ~ * ~ J
I |
* * * * j
i
+ ***
! 2.03 '
| |
3. 1 1 '
1
1 . 82

1
1
1

1 1
1 1
| |
1
i
i

J 3.96 * j
1
1
1


1

1 1
i

1 1.56*1
1


1
1

1 1
1 1
i
i

! 2.54*'
1
1











1

j |
i

1 |
j

* * * ~
1
1
* * * *
J ~ * * * j
***~ j
* * * *
j 1.47 !
2 .00 j
2 .50
* * * *
1
1
* * * *
1 * * * * 1
1 |
~ * * * i
1
*** +
' 1,67 !
1 . 16 |
2.51
* * ~ *
1
1
1
* * * *
| * * * * |
1 I
i
i
~ ***
1 0.BB 1
1 1
1 .72|
1
1.00

1
1
1

1 1
1 t
I |
i
i
i

1 1
j 1.65*|
1
1
1


1
1

1 1
i

I 2.03*|
1
1


1
1

t 1
I 1
i
i

1 0.81*!
1
1










* ~ * *
1
~ ~ * *
1 * * * * I
~ ~ * * i
* * * *
! 2.78 I
4 .021
36. 30
~ * * *
1
1
** **
1 * * * * |
**** '
* * + +
1 2.19 i
1 . 191,
35 . 86
****
1
1
* * * *
1 * * * * j
* * * * •
+ * * *
j 1 .27 |
3.38J
1.01

1
1

1 1
1 1
i
i

' 2.99*1
1
1


1
1

1 1
1 1
i
i

| 1.86*|
1
1


1
1

1 1
| |
i
i

! 1 •611
1
1










*¦** +
I
1
I
* * * *
1 1
1 * * * * 1
i
# * * * i
* * + *
1 1
! 1.96 j
1
4 . 98j
3 . 59
* * * *
1
1
*<¥**
1 1
j * * * + |
* * * * i
* * * »
1 1
1 1 . 1 S I
* ~ * *
7 .07
****
1
1
* * * *
! * *~* [
*»** j
* * * ~
! 1 -70!
* ~ * *1
1
0.51

1
1

i i
i t
i
i

' 3,50*!
1
1


1
1

i i
i i
i
i

j 8 . 5 1 i
1
1


1
1

i i
i i
i
i

j 0.41*,
1
1

00
~SPECIAL REPORT DATED OCTOBER 12, 1977
VALUES OF **** REPRESENT DATA NOT AVAILABLE

-------
TABLE 2. 
-------
A1 PROD
CHLA
ASMA
A2 PROD
CHLA
ASMA
A3 PROD
CHLA
ASMA
A4 PROD
CHLA
ASMA
A 7 PROD
CHLA
ASMA
A8 PROD
CHLA
ASMA
A9 PROD
CHLA
ASMA
All PROD
CHLA
ASMA
TABLE 3.
1978 PRODUCTIVITY, CHLOROPHYLL A, AND ASSIMILATION RATIO A
(_n
O
JAN
FEB
MAR
APR
MAY
JUNE
JULY AUG
SEPT
OCT
NOV
DEC.
1 .05
1.04
1.01
3.211
1	.49 j
2	. 1S '
*****
* ** * *
*****
.94
.53
.77
0.70
0.65
1 .08
2.56
2.BB
0 .89
2.51
2.76
0.91
0.59 |
0.48 j
1.23 {
2.99
0.73
4.10
4.72
2. 04
2.31
5 .27
1 .09
4 .83
13.031
2.53,'
5.15'
1.98
1.16
1.71
3.84
1 .69
2.27
3.26
2. 16 j
1.511
	I _
5.42J
3.241
1 .67|
0.68
1 .29
0.53
2.81
2.68
1 .05
. 15
.36
.61
.08
.96
.21
6.51
3.37
1 .93
7.12
3.79
1.88
10.10
3. 75
2.69
6.16
4 .04
1 .53

2.98 J
2.14 I
1 .35 |
20.09
6.46
3.1 1
7.7 2
4.77
1.62
9.11
8.45
1.08

5.94 I
2.18|
I
I
2.73
9.93
2.77
3.59
	
9.20
4.06
2.27
	
11 .06
4 .38
2.53
7.20
3.55
2.03
5.73
3.12
1 .84
	1
5.73J
1. 191
4.82|
	I
12.85|
1 .47}
8.741
	1
8 . 39j
1.24*
6.77|
I
	1
16.97J
2.271
7.48j
	I
4 .69!
1 .OA1,
4.51}
	1
7.67!
1 . 19]
6 .451
I
	!
I
6.8 31
1 . 53i
M
HI
O
CTi
3.06
1 .20
2.55
6.031
2.00'
3.02'
4.20
3 .72
1.13
11
2
3
.42
.86
.99
7.98
4 . 14
1.93
13.08
5 .37
2.44
I 10. 14
J 8.21
I 1 .24
4.47 j
3.38 |
1 .32 I
8.6 7
2.63
3. 30
12.81
2.93
4.37
19.26
13.62
1 .41
	1	
0.39{14. 39
4.151 3.65
3.86
2.13
1.21
1 .76
13 . 9 1 j
1.92j
4 .641
3.73
2.32
1.61
.72
.09
.41
8.89
4 .06
2 . 19
13.75
5.21
2.64
2.83
5. 76
I 0.49
1
0.09i
I
10.16
3.78
2.70
37.67
5.79
6.51
0.85
1.03
0.83
1.66J 1.06
1.431 1.18
1.1 ej 0.90
1.39
1 .76
0 .79
1 .65
2.39
0.69
3.81
5.86
0 .65
'I	
• 2.72
4.70
} 0.58
I	
	—T-
5.23j
3.73|
1 .40|
6.24
4 .06
1 . 54
6.24
5.12
1 . 22
17. 82
4 .05
4.40
1.40
1.03
1 .36
I
1.111 2.06
0 . 78| 2.5 3
1.42J 0.81
1 .70
1 .44
1.18
3.62
2.72
1 .33
1 .88
2.54
0.74
I 2.36
{ 2.42
! 0.98
2 . 83 |
3.01}
0.94 !
7.85
3. 79
2.07
5 .33
2.91
1.83
|26 .01
8.27
3. 15
I
& * * * *
* * * * 41
*****
8.07] *****
1.60{ *****
5.041	*****
I
4 .31
1.67
2.58
2.79
1.61
1.73
8.68
I * * * + *
8.46 I *****
1.03 | *****
5.59J
9.011
0 . 62 j
	L
11 .78
2.40
4.91
21.84
i 2.90
j 7.54
67.75
13.26
5.11
4.46!
VALUE OF *****REPRESENT DATA NOT AVAILABLE
-------
TABLE 3. (CON'T)
1978 PRODUCTIVITY, CHLOROPHYLL A, AND ASSIMILATION RATIO A (CON'T)
A12
A13
A14
A15
A16
A17
B8
39
CI
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
PROD
CHLA
ASMA
h
JAN
FEB
MAR
APR
MAY
JUNE
JULY AUG
SEPT
OCT
NOV
DEC
1 .31
1 .08
1.21
1 .65
0.84
1 .96
h
1 .40
0.90
1 .56
3.67
1 .78
2.06
2.14
0.82
2.61
2.31
1 .70
1 .36
2 .64
1 .22
2.16
* * * *
+ * * *
~ * * *
0.40
0.38
1.05
2.24 j 6.15
1.75 I 3.02
1.28 ! 2.04
2.29 I 1.79
1.98 1 3.64
1.16 { 0.49
3.72 | 5.23
2.25 J 2.47
1.65 12.12
	1	
6.33 J 7.12
2.09 J 3.79
3.03 | 1.88
6.98 j 6.22
1.98| 3.76
3.53 j 1.65
5 .65 I
3.21 j
1 . 76 [
2.53
1 .49
1 .70
3.27 |
1 .65 |
1.981
7 .64
1 .59
4.81
3.76 j
2. 12 J
1 .77 i
5.52
3.72
1.48
	T-
* ~ * * |
**~* J
+ * **
* * * *
4
4.30 j 3.43 j 9.10
1.69 I 1.74 I 2.40
2.54 j 1.97 ! 3.79
1.58 I 1.49
1.52 ' 1.23
1.04 J 1.2 1
1.97 [ 2.03
1.57 { 0.89
1 .25 I 2.28
I
3.46 { 4.80
0.95 I 3.42
3.64 j 1.40
3.20 j 4.30
1.32 ! 3.26
2.42 ! 1.32
2.06 I
3.40 j
0.61 J
1 .05
1.61
0.65
4.76 ,
1 .31 j
3.63 I
4.28
2.30
1 .86
	T
5.71 j
* ~ ~ ~
3.87
2.07
1 .87
~	* * ~ j
*	* ** ]
***+1
I
* * * *
****
****
3.59
2.60
1 .38
8.19
2.86
2.86
6.34
5.05
1.26
5.67
3.27
1 . 73
3.80
3.11
1 .22
3.82
3.10
1.23
1.12
4.05
0.28
****
* ~ * *
** * *
7.17
5.51
1 .30
0. 94
3.64
0.26
6 . 66
5.70
0.17
2.65
3.36
0.79
2.54
4.34
0.59
3.01
4.88
0.62
h
3.30
3.77
0.88
5.32
3.95
1.35
****
****
*
	T-_	T,	
2.13 |2 1.58 J 4.83
1.59 | 5.131 2.82
1.34 j 4.21j 1.71
	i.	4-	
61 .56 |19 .22|
16.43} 2 . 4 8j
3.75 I 7. 75
-A-
1.51 |19.48| 4.09
1.45 j 7.34 j 2.50
1.04 ! 2.651 1.64
2.23
1 .54
1 .45
j 23.54 J
! 4.171
6.86
2.88
-f
5 .6 5 i 2.38
	1	
2.47 j 17.00 j 6.11
1.64 | 2 . 871 6.07
1.51 j 5.92 j 1.01
	J	4-	
0.97
2.99
0.32
110.10)11.21
j 2.90 J 3.48
! 3.48! 3.22
0 .33 j 21 .58] 7.86
1.91 { 5 o13 j 2.49
0. 17 I 4.211 3.16
2.47
1 .82
1 .36
! 9.19!
4
* * + *
** + *!
9.83
3.86
2.55
1 .66
1.74
0.95
I 34.661
j 11.83j
| 2.93.
6.48
3.95
1 .64
**~*
****
****
* * * f|
**«*l
***«!
** * *
* * ~ *
* ~ * *
5.42 }l3. 17
2 . 00 I 2 . 00
2.71 | 6.59
15.79 03.78
2.32 j 2.05
6.81|6.72
	JL
37.89 bl .27
7.39 j 1.53
5.1313.90
5.99 {14.72
4.281 1.65
1 . 4 0 { 8.92
6.82tt*-32
1 .34 I 1 . 79
5.09| 8.00
	J	
19.54(11.28
7. 32,' 1.66
2.67J 6.50
	j	
28.51 |l 1 .82
6 . 08 j 2.34
4.6915.05
1
¦r
*****1 ** *
»
* * * * * j
*****
***
I ***
	L_
-------
Intentionally Blank Page
-------
IID
153
CHANGES IN ZOOPLANKTON
IN OUTER LOS ANGELES-LONG BEACH HARBORS
1972 - 1978
INTRODUCTION
The zooplankton of Los Angeles-Long Beach Harbors is com-
posed largely of calanoid copepods, cladocerans, chaetognaths,
larvaceans, and the larvae of fishes and benthic invertebrates.
Acartia tonsa, a calanoid copepod, is the most abundant species.
This copepod is a hardy animal, able to tolerate relatively
high temperatures and low salinities (AHF, 1976; Jeffries, 1962).
Many zooplankters are generalist feeders, consuming flag-
ellates, detritus, and bacteria. The percentage taken of each
food source differs according to the assimilation rates and size-
related filtering efficiencies of the zooplankters (Saunders,
1970; Brooks, 1970). The proportion of each species in the pop-
ulation as a whole may vary seasonally or spatially according to
the food supply.
Enrichment by fish wastes or sewage favors two foods of zoo-
plankton: phytoplankton and bacteria. Diatoms and dinoflagel-
lates can increase markedly in areas of nutrient enrichment, sup-
porting large populations of zooplankton (Marshall, 1947; Parsons
et at., 1977). In more turbid waters, heterotrophs in the water
column may convert about 70% of the daily input of metabolizable
carbon into particulate organic carbon (Sibert and Brown, 19 75).
Rod-shaped bacteria may be abundant on particulate organic matter
(Ferguson and Rublee, 1976; Sullivan et at. , 1978). The zooplank-
ton feeding on these foods may either remain in the open-water
food chain or eventually settle out to become food for benthic
invertebrates and demersal fishes.
In order to assess the effect on zooplankton of beginning
secondary treatment at the Terminal Island Treatment Plant,
the plant's operations and the inclusion of secondary treatment
of cannery effluents, it is necessary to determine the character-
istics of zooplankton populations in and outside the area of in-
fluence from a historical perspective. To accomplish this Har-
bors Environmental Projects data from 1972-1978 were used.
METHODS
Plankton data from stations Al, A3 and A7 (outside harbor;
mid-outer harbor and area of TITP influence, respectively) were
analyzed from January 1972 to the present. Stations are shown
on Figure 1. Those species looked at in detail were the three
dominant copepods, Aaavtia tonsa, Paraaalanus parvus, and Cory-
aaeus angliaus and the three dominant Cladocera, Podon polyphe-
moides, Evadne nordmanni, and Penilia avirostris . Also included
-------
154
IID 2
in this study are the total zooplankton concentration and the
diversity index (Shannon-Wiener) of copepods and cladocerans
through the years of sampling. These dominant species, as well
as the total zooplankton concentration and diversity index, are
shown in Figures 2 through 13. Figures 2 through 4 and 6 through
8 show the concentrations (number/m^) of copepods and cladocerans
respectively, collected from January 1972 through September 1977
using surface horizontal tows. Figures 5 and 9 show the concen-
trations of the same species, using vertical tows from bottom to
surface from October 1977 through December 1978. Total zooplank-
ton concentration and the diversity index of copepods and clado-
cerans are shown from the same sampling dates and methods in Fig-
ures 10 through 12 and 13, respectively. While the horizontal
and vertical plankton sampling do not yield similar concentra-
tions (vertical tows showed higher concentrations than horizon-
tal tows), as evidenced by studies comparing the tow techniques,
similar zooplankton trends (zooplankton maxima) should be evident
and relative differences between the stations should be similar.
RESULTS•
Aaavtia torisa
The dominant zooplankter, Aaartia tonsa, generally compri-
ses over half of the total zooplankton concentration. Through
the years of sampling, this species generally shows a low summer
concentration, with a depressed mid-winter period. Concentra-
tions are usually high in late fall-early winter and in late
winter-early spring. This was the typical pattern from 1972 to
to early 1974, the only deviation being when A 7 showed a peak in
the summer of 1972. The winter of 1974-75 and 1975-76 showed
some deviation from the former pattern by the absences of an
early spring and late fall peak, respectively. There was also
an absence of a late fall peak in 1976, but the late winter-ear-
ly spring peak of 1977 had an unprecedented high concentration
of about 18,600 A. tonsa/m^ at stations A3 and A7, with a low
concentration (about 500 A. tonaa/wr) for Al. The months of this
exceptional concentration of A. tonsa (March-April) peaked at a-
bout the time that the Terminal Island Treatment Plant (TITP)
converted to secondary treatment in April. The bloom probably
was underway before conversion. It followed an unusually warm,
dry winter that had some two inches of rain in late March. The
extraordinary bloom of this single species made the zooplankton
nearly axenic as evidenced by the unprecedented low diversity
among copepods and cladocerans for a single month at stations
A3 and A7, nearest the outfalls.
As diversity is advantageous for the stability of an ecosys-
tem, anything which would cause the diversity to decline would
be disadvantageous. Thus the initiation of secondary treatment
may have been detrimental to the zooplankton ecology at that time;
-------
IID 3
155
however, the A. tonsa concentration returned to near normal con-
centrations by summer 1977, as did the diversity index. By Sep-
tember, TITP started processing the effluent from one cannery
and by January 1978 all cannery effluent had entered TITP. It is
this winter of 1977-78 that, for the first time since 1972, nei-
ther late fall nor early spring A. ton3a maxima were evident,
which might be linked to the lack of effluent being discharged
from the canneries. An A. tonsa peak, however, recurred in the
fall of 1978 at station Al outside the harbor and at A3 in mid-
outer harbor, but not A7, near the treatment plant outfall, a
station which had participated in the winter-time peaks in all
pre-secondary treatment years.
It is interesting to consider the mean concentrations of
A. tonsa at the above three stations during pre- and post- can-
nery effluent treatment. These data, as well as other copepod
cladoceran species data, along with total zooplankton and diver-
sity indices of copepods and cladocerans, are shown in Text Ta-
ble 1. It can be seen that prior to cannery effluent treatment,
the mean A. tonsa concentration was greatest at A7 and A3, with
only about half those concentrations at Al. Following the can-
nery treatment, the mean A. tonsa concentrations indicate that
the highest concentrations occurred at A3 and Al — over three
times greater than at A7. This depressed concentration at A7
may be the result of secondary treatment processing; however,
it must be remembered that the post-cannery treatment mean is
based on fewer samples, and on samples which were collected by
vertical tows, so that they are not directly comparable.
Text Table 1. Pre- and Post-Cannery Treatment
Zooplankton Concentrations
Pre-Cannery Treatment (January 1972-September 1977)
A.tonsa
P. parvus
C. angliaus P.poly-
E.nord-
P.avi-
Total
Species


phemo-Ldes
manni
rostris Zooplank.
Diversity
Al 577
598
93 403
943
16
3600
1.26
A3 1037
230
56 382
412
62
2528
1.07
A7 1133
150
36 452
286
7.8
2454
0.99
Post-Cannery Treatment (October 1977-December 1978)


Al 1155
788
335 41
216
946
5514
1.56
A3 1398
578
101 41
58
394
3357
1.17
A7 370
202
38 60
14
36
1377
1.29
Paraaalanus parvus and Coryaaeus angliaus
The next two most abundant copepod species are the calanoid,
Paraaalanus parvus, and the cyclopoid, Coryaaeus angliaus, which
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156
IID 4
make up 10% and about 1.6% of the Los Angeles-Long Beach Harbor
zooplankton, respectively (AHF, 1976). The concentrations of
these species at stations Al, A3, and A7 over time are shown in
Figures 2 through 5. There seems to be no clear seasonal trend
for either of these species. The only major bloom of P. parvus
at stations A3 and A7 occurred in the winter, 1972-73. Other
than that peak, P. parvus showed peaks at station A7 greater
than 500/m^ only immediately after TITP converted to secondary
treatment (April and May, 1977). Whether the lack of sewage and
cannery effluent contributed to these peaks is unknown, since
the overall ratio of Al to A7 P. parvus concentrations (Text
Table 1) remains unchanged from pre- to post-cannery treatment.
P. parvus has been shown to be the most ubiquitous species
(AHF, 19 76) and thus may be the species least affected by pertur-
bations in the environment.
Coryaaeus angliaus is another species which showed only one
major bloom during the seven years of sampling. This occurred
in the spring of 1974. This species did not seen to show any
stimulatory or inhibitory effect by the initiation of secondary
treatment for-sewage or cannery effluent, although the effect on
total populations cannot be predicted for the long term.
Cladocerans
Cladocerans form the next most dominant group behind cope-
pods. Podon polyphemoid.es and Evadne nordmanni are che two dom-
inant cladoceran species comprising 11.1 and 4.7 percent of the
zooplankton, respectively (AHF, 1976), with Penilia avircstvis
a distant third of minor importance. The changes in concentra-
tions of the species over the seven years of sampling is shown
in Figures 6 through 9.
Podon polyphemoides
Podon polyphemoides is a species that had two major blooms
during the seven years of sampling. Those occurred in the spring
of 1972 and 1976. Another bloom may have been in the making in
the spring of 1977, as evidenced by the substantial increase of
P. polyphemoides at A7. This bloom may have been cut short by
the conversion of TITP to secondary treatment, since never before
had there been such an intense increase without its lasting more
than one month.
Following the first cannery effluent entering into TITP in
September 1977, all three stations showed extremely low abundance
of P. polyphemoides. After the last cannery was connected to the
TITP (January 19 78), there was a total absence of this species
in the water column up to June 1973. Such an absence occurred
one other time, in the spring of 1975, but for a much shorter
period of time. Since P. polyphemoides was absent from all sta-
tions rather than just A7, it would tend to suggest that the ab-
sence might not have been a result of cannery effluent treatment.
It should be noted, however, that this absence was broken with
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IID 5
157
the return of P. polyphemoid.es to the water column following the
malfunction of TITP in June. After September, when TITP was op-
erating again, this species persisted in the water column through
December. Text Table 1 shows the drastic drop in P. polyp.hemoi-
des concentration after TITP processing of cannery effluent be-
gan. It also indicates that this species was evenly distributed
and was not more or less abundant at any one station, either be-
fore or after TITP cannery effluent treatment.
Evadne nordmanni
Evadne nordmanni showed major blooms timed similarly to
those of P. polyphemoides, both in the spring of 1972 and 1976.
Evadne nordmanni shows no consistent seasonal trend, and as in
the former cladoceran species, it, too, showed sparse populations
present at all stations following the secondary treatment of can-
nery effluent. This low population level continued at all sta-
tions until October 1978, when Al, outside the harbor, showed a
small bloom. Text Table 1 shows that E. nordmanni also experi-
enced a decline in concentration as well as a change in numbers
collected at station A7 as compared to Al. Prior to secondary
treatment of cannery effluent, the Al to A7 ratio was 3:1, while
after treatment the ratio was 15:1. While the values may not be
significant because of the fewer samples after treatment, it may
indicate a trend toward greater sparsity of this species at the
station (A7) most impacted by the secondary treatment of cannery
effluent.
Penilia avirostris
This relatively minor member of the zooplankton is absent
most of the time; however, when it is present, it is often very
abundant. The only real abundance in the past six years prior
to effluent treatment occurred in the fall of 1976. After treat-
ment, however, a tremendous bloom occurred in the fall of both
1977 and 1978. These blooms were restricted primarily to Al and
A3, and thus may not be related to the cannery effluent treatment
which might be manifested primarily at station A7, as compared
with Al and A3.
Total Zooplankton and Diversity Index
The concentration of total zooplankton and the Shannon-
Wiener Diversity Index for copepods and cladocerans collected over
the seven years of sampling stations Al, A3 and A7 are shown in
Figures 10 through 13. The seasonal characteristics of total
zooplankton are primarily functions of the already discussed
dominant species of copepods and cladocerans. Text Table 1
shows, as would be expected from the previous discussion, that
there was a reduced concentration of total zooplankton at A7 as
compared to Al, after secondary treatment of cannery effluent was
initiated. Prior to treatment, the A1:A7 ratio was 1.5:1, while
after effluent treatment the ratio changed to 4:1.
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158
IID 6
Species diversities of copepods and cladocerans show consid-
erable variability between stations and over time; generally they
are higher at A1 outside the harbor than inside, and at station
Al there appears to be greater species diversity during the win-
ter months than during the summer months. One period of very
low diversity was observed at A3 and A7 in April 1977 with a
bloom of Aaartia tonsa, during the period when secondary sewage
treatment was initiated.
Text Table 1 indicates a decline in diversity from outside
the harbor (Al) to inside (A7) before cannery effluent treatment/
as might be expected. After treatment, however, while Al still
had the greatest diversity, station A7 seemed to have been im-
proved as compared to A3, since A3 then had the lower diversity.
This may indicate that cannery effluent treatment may have im-
proved the environment at A7, but it would perhaps have de-
creased the food available at A3, although the results are not
conclusive.
Distribution of Total Zooplankton
Mean concentrations of zooplankton in 1973-74, shown in
Figure 14 (AHF, 1976), indicate that outer Los Angeles Harbor
was not an area of high concentrations; however, the same area
was indicated as one of high fish concentrations in 1972-74, and
predation may well have been the controlling factor except, per-
haps, at those stations within the shallow water area closest to
the outfalls (A4, A7, All). Fish trawls were not taken in the
shallow water area.
In spite of the increase in populations in 1978 at several
perimeter stations (A2, All, B8, B9), the mean total concentra-
tions decreased at A7 (Figure 15). The 1978 data for other areas
of the harbors have not been analyzed as yet, so it is not known
whether changes have occurred in the B, C and D stations as well.
CONCLUSIONS
The milestone events during conversion of waste treatment
from primary to secondary and inclusion of cannery wastes in
TITP can coincidentally be seen in variations in numbers of the
dominant zooplankton in the harbor as compared with a station
outside the harbor. Although the variations cannot be proven to
be due to the changes, the coincidences are notable.
The copepods probably feed largely, and perhaps preferen-
tially, on bacteria on organic detritus; therefore the more than
thirty-fold decrease in total marine bacteria plus a four- to
seven-fold decrease in phytoplankton assimilation would have had
an effect. Copepods apparently are able to select enriched par-
ticles from unenriched particles of the same size (Poulet and
Marsot, 1978).
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IID 7
159
The fact that total concentrations are not greatly
increased might be related to the large drop in fish predators
discussed elsewhere, leaving about the same standing crop even
if the gross production was much reduced. Replenishment by
tidal exchange is also important, but is largely unquantifiable.
Changes in the method of collection between 1974 and 1978
preclude direct comparison of total zooplankton.
The so-called "zone of inhibition" previously identified
(Soule and Oguri, 1976) near station A7 was an area of very
high nutrient levels, but the turnover of the nutrients was
postulated as contributing to the "zone of enhancement" that
included most of the outer harbor. The perimeter of the "zone
of enhancement" may now have moved much closer to A7, resulting
in improved diversity there and larger populations in the
outermost stations of the area (A2, B8, B9). However, both A7
and Al decreased substantially in total populations. Since
no further monitoring is planned after December 1978 by the
City of Los Angeles, it will not be possible to say whether
the apparent changes represent long-term trends.
LITERATURE CITED: See Section Vi.
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160
IID 8
-B6J
WILMINGTON
LONG BEACH
BSj
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Harbors Environmental Projects
University of Southern California
Figure 1.
Zooplankton Analysis of Selected Stations
-------
IID 9
161
.5565
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•3154
3*96
2B00-
n
I
1000-
9625
9000-
6O00-
4000-
200O
1000-
1973
V72
Figrn 2 Dominant copepod concentrations. Jan. 1972 - D*c 1973
Station A1	ja	,A7	
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162
IID 10
1
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woo-
soo
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1876
Flgun 3. Domintm eopeood concwitrdiona. Jan 1974 - Dec 1978
SMttofl A1	 A3	,A7	
-------
I ID 11
163
Rgun« Dominant cope pod concentralions. J«n 1978 Sap! 1977
Station A1	>3	,A7	
-------
164
IID 12
too.
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Figur* S. Oomiwtt eopapod concant rat ions. Oct t977 - Oac. TS78
Station Al	 M	JO	
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IID 13
165
600-
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1 2000-
1600-
600-
4000-
3600-
Ftgur* A, Dominant dtdocsran ooncantraliona. Jmn. !972 - Doc. 1873
Station A1	A3	 ja	
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166
IID 14
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Plgura 7. Dominant cladocaran concentrations, Jan. 197* • Oac. 1878
Stationa *1	, A3	, A7	
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-------
168
IID 16
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Figure 9. Dominant cladoceran concentrations. Oct. 1977 - Dec. 1978
Station A1	.A3	,A7-
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IID 17
169
1.5-
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28.939
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12.000-
10000-
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I	1972	I	1973	t
Figur* 10. Total zoopltnktcn cone. ¦. Cooapod and cladocaran divaratty
Jan. 1972-Oae. 1973 Station. Al	,A3	,AJ	
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170
IID 18
£
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Figure 11. Total zoopiankton cone. ; Copapod and cladocaran divaraity
Jan. 1974-Dae. 1975 Stationa A1	 ,A3	,A7	
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IID 19
171
2.0
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J«n. 1878 - S«p. 1977 Station* At	,A3	,A7	
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172
IID 20
1.5-
1.0-
0.5-
8000-
6000
4000-
2000
1978
Figure 13 . Total zooplankton conc. Copepod and cladoceran diversity
Oct. 1977 • Dec. 1978 Station A1	,A3	,A7	
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REGULAR MONITORING STATIONS
1973 - 74
WILMINGTON
LONG
BEACH

SAN
PEDRO
ANGELES \
LONG
BEACH
HARBOR
LEGEND
S 1500
1500< • s 2000 MEAN CONC
2000 < • S 2500
2500 < • S 3000
3000 « * S 4000
>4000
A	SERIES 2354
B	SERIES 4420
C	SERIES	2308
D SERIES 2372
PT. FERMIN
SCALE IN MILES
U.s.c.
Figure 14. Mean Spatial Distribution of Total Zooplankton (Source: AHF, 1976)
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174
IID 22
WILMINGTON
LONG BEACH

Pier J
PEDRO
Legend
' £ 1500
1500 < • £ 2000
2000< * £ 2500
2500 < • £ 3000
3000 < • £ 4000
> 4000
Harbors Environmental Projects
University of Southern California
Figure 15. Mean Concentrations of Total Zooplankton
in the Outer Harbor, 1978
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HE
175
CHANGES IN BENTHIC FAUNA IN OUTER LOS ANGELES-LONG BEACH
HARBORS, 1972-1978
INTRODUCTION
Benthic organisms have been sampled extensively in the
outfalls area of outer Los Angeles Harbor since 1971 by Harbors
Environmental Projects, and results of studies through 1974
were published in part in 1976 (AHF, 1976).
In the 1973-74 period, 43 stations were sampled quarterly
and results showed that for the entire harbor the average
sample contained 28 benthic species and 1404 individuals per
1/16 m2. The range of variation was high, with species numbers
from 1 to 60 per sample, and abundances from 2 to 5000 per
sample. The latter number becomes 80,000 organisms when cal-
culated to the square meter of surface, an extremely rich
benthos as compared with other local soft-bottom areas (Soule
and Oguri, 1976; Word, Myers and Mearns, 1977). Other studies
on biomass and simulated recolonization of dredged areas
indicated that biomass averaged 20-30 gm/m2 in the outer
harbor near the breakwater, 200 gm/m2 at stations nearest
shore, and 500 gm/m2 in the central portion out from the cannery
and sewage outfalls area (the zone of enrichment) (Soule and
Oguri, 1976a, b).
A quote from the Master Environmental Setting for the Port
of Long Beach (1976) with regard to the importance of benthic
populations and plans to dredge and fill the central enhanced
area reads as follows:
"The importance of benthic animals to the entire
food web of the harbor is not well recognized. Poly-
chaete worms are the largest number of harbor benthic
organisms. They filter organic detritus and bacteria
out of water, or consume them from sediments when
feeding. Polychaetes, in turn, furnish a major food
source for large harbor fish populations of a number
of species. If both ports completed their Southwest
Basin dredging and filling, an estimated 8.5 x 10s grams
(850 tons) of organisms would be lost by burial; dredg-
ing would destroy an additional 6.8 x 108grams (680 tons).
"Since large polychaetes may weigh less than 0.1
gram, that represents 15.3 billion worms. In one fish
stomach examined recently, 600 worms were found, so
the direct consequences for those fish feeding on
benthic polychaete worms can be estimated as 25 million
'fish meals.'"
The area of enhancement is dependent upon the existence
of a large, slow-moving gyre in the outer harbor directly south
of the outfalls area which apparently circles clockwise on the
-------
176
HE 2
surface and counter-clockwise beneath (Soule and Oguri, 1972;
Robinson and Porath, 1974). The surface gyre has been verified
and simulated by the U.S. Army Engineers Waterways Experiment
Station model at Vicksburg, Miss. (McAnally, 1975, 1976); it
is figured in section IVA. The harbor gyre system facilitates
distribution and assimilation of the organic nutrients, and
contributes to oxygen levels by mixing, which accounts in
large measure for the health of the harbor.
METHODS
The locations of the benthic stations sampled in the
harbors are given in Figure 1, along with the stations sampled
in prior years in various studies. The box on the map outlines
the TITP study area; station A7 is near the outfalls location.
Harbors Environmental Projects has published and unpublished
records for stations A1-A8 beginning in 1971 with studies for
Pacific Lighting Corporation's proposed LNG terminal.
Sediment samples were taken from the RV Vantuna in 1977-78
using a stainless steel Reinecke box corer. The corer is a
modified spade corer which takes a sample with 1/16 m2 of sur-
face and up to 30 cm in depth. On board, two 100 cc subsamples
for chemical analysis and for grain size analysis were taken
from the middle surface of each core and frozen immediately
for later laboratory techniques. The upper half of the core
was then taken and washed with running sea water through a
0.5 mm screen. Material retained on the screen was fixed
immediately in formalin-sea water and later transferred to
isopropyl alcohol for sorting and identification. In locations
too shallow for the Vantuna (at A7) the RV Golden Vest was used
to operate a stainless steel Campbell grab sampler (similar
to a Van Veen), which takes a 1/10 m2 surface sample. Care
was taken to obtain the chemical samples from the surface of
the box core where it would not have been touched by metal. Since
the surface sediment is undisturbed with the Reinecke corer,
it is the preferred gear. In addition to the results discussed
below in this section, section IV contains extensive discrim-
inant computer analysis of benthic data.
Data gathered on the 1977-1978 benthic samples were
compared with the data previously gathered, beginning in 1971.
In addition, stations were compared concurrently according to
locations (spatially) to seek indications of differences in
the harbor environment and in seasonality.
RESULTS AND DISCUSSION
The baseline on benthic sampling beginning in 1971 first
condensed to annual means at all stations sampled in each year,
for both numbers of species/m2 and numbers of individuals/m2
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iie 3
177
(Figure 2). The means for each station have been calculated
in Table 1 for all sample years (across the page) and for
all stations in each year (vertically).
Annual Mean Trends in Diversity and Populations
In the benthic data plotted as annual means, there was
very low species diversity (richness) and numbers of organisms
per m2 (abundance) in 1971. This no doubt reflects the com-
paratively poor harbor environment that existed prior to
enforcement by the RWQCB of prohibitions on oil refinery and
other toxic wastes in September 1970 (Reish, 1959; Crippen and
Reish, 1969).
Reish (1971) documented the dramatic change in the inner
Los Angeles Harbor as anoxic conditions began to- disappear.
Fish Harbor, in outer Los Angeles Harbor (stations A5 and A6)
also was anoxic from dumping of fish wastes, process and non-
process water; the wastes were diverted in 1970 to outfalls
in the A7 area near TITP. The large increases, to 1972-74
levels, seen in both species diversity and abundance are probably
due to the intensive enforcement effort, which allowed exten-
sive new colonization to occur. Such an increase will usually
be followed by a slight drop before stabilization of species
and populations occur (Soule and Oguri, 1977) in normal suc-
cession. Thus, the principal trends in 1971-1973 were steep
climbs in both numbers of species and numbers of individuals.
The curve leveled off in 1974 for numbers of species but began
to decline for numbers of individuals.
The principal trend since 1974 in the Los Angeles Harbor
area has been a decline in total abundances (numbers of indi-
viduals) of benthic organisms. The trend in species diversity
was upward through 1976, followed by a moderate decline in
1977 and a steep decline in 1978.
Site Specific Trends in Diversity and Populations
In order to examine the trends in different benthic loca-
tions, several stations were selected according to their
distance from the TITP and cannery outfalls. Station Al is
located outside the harbor, whereas A2 is in the seaward area
of the outer harbor, and station A3 is about halfway between
A2 and A7. Station A7 is closest to the outfalls, being
between the cannery outfalls on the west and TITP outfall to
the east. Station A2 is located at one of the harbor Coast
Guard buoys which was moved from A2a (Figure 1) to A2 in
March 1973. Hence there was some difference in the substrate
and species found in 1972 to 1973 data. This apparently did
not affect other biological data since water column samples
are not as static as benthic samples are.
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178
HE 4
The data for the specific sites mentioned are presented
in Figures 3-6, on a seasonal (approximately quarterly) as
well as an annual basis. Numbers of species and numbers of
organisms per mz for each station are plotted and the trends
discussed below.
Station Al. While station Al is outside the Los Angeles
Harbor entry (Angels Gate) and more subject to ocean conditions,
it is still tidally influenced by the harbor. Terrigenous
nutrients are carried by the natural flow of fine sediments
down slopes to deeper waters at all river and bay entrances
not blocked by sills or sand bars. The nutrients support the
phytoplankton, algae, marine plants, microheterotrophic
bacteria, and protistans on which benthic organisms feed.
Thus the benthic zone of enhancement from the enriched harbor
actually extends outside a short distance, especially for
benthic organisms. The nutrient-poor (oligotrophic) character
of most of the ocean waters may be encountered only past the
Channel Islands in some cases, because of the terrestrial
input of nutrients. It is because of terrestrial inputs that
coastal inshore biota are the richest around the world. Even
nutrients in upwelling areas were carried first from the coasts
down into deep canyons before being recycled by upwelling.
The years 1972, 1973 and 1974 were peak years for benthic
organisms outside the harbor at Al (Figure 3) as well as inside
the harbor. Although all stations showed a precipitous drop
in abundance beginning in mid-19 74, it was least dramatic at
Al where benthic populations have always been smaller.
In the spring of 1976 and 1977 the expected late winter-
early spring increases in numbers occurred but were of rela-
tively small magnitude at Al. The slope of the abundance
curve has been down, except for seasonal variations, since
February 1975. The use of DAF treatment by the canneries
began, coincidentally, in March 1975, reducing the nutrient
load greatly.
Species diversity rose steeply over the long term, from
1971 through 1976, at Al, but with great seasonal fluctuations.
Species diversity then dropped very steeply after December
1976 through 1978 and showed no sign of recovery other than
seasonal fluctuation. Thus both species abundance and species
diversity appear to have suffered a long-term decline at Al,
in 1977 and 1978.
Station A2. The outer harbor station A2 (Figure 4) showed
about four times as many organisms in August 19 74 as Al
(Figure 3). Station A2 has shown about a tenfold decline in
abundance overall between May of 1974 and the end of 1978,
but there were larger intervening seasonal peaks than those
at Al. An unusual summer peak in 1976 and 1977 occurred at
stations A2 and A3 (Figures 4, 5) which did not occur at Al.
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HE 5
179
Station A3. The net slope of the curve for abundance at A3 is
generally downward from a peak in 1972-73, but it was not as
steep as that for A2. The species diversity seemed to be fair-
ly stable at A2 and A3, although they show large seasonal fluc-
tuations. Peaks in diversity occurred in the summer of 1976,
1977	and 1978 at A2, A3 and A7 but not at Al.
Station A7. At station A7 (Figure 6) during the summer, fall
and winter of 1976, the abundances declined severely but recov-
ered slightly in June 1977. Numbers dropped again in Septem-
ber 1977, recovered in the winter, dropped in April 1978, re-
covered in July, and dropped again in October 1978. Note the
lower scales on the A7 graph. Station A7 was greatly influenced
by the outfalls. When DAF units were unstable, or were using
large quantities of alum in 1975-76, when TITP construction
created unusual conditions, such events were reflected in the
benthic populations. It is possible that drought conditions
in the winter of 1975-1976 also affected the area by reducing
flushing. Chlorination was in effect from March through August
1978	and may have had a deleterious effect (Oliver and Carey,
1977; Emerson, 1976). Also, about 30 inches of rain fell in the
three months of 1978, which would affect plankton more than ben-
thic organisms. The diversity peaks in the harbor were lower
near the outfall at A7 in 1976 and 1977; however, in 1978 the
peaks in both abundance and species diversity were comparatively
higher than at A3.
The 1977 and 1978 variations are compared in Figure 8 for
stations Al, A2, A3, A7 and A12 and in Figure 9 for A4, A8,
A9 and All. In Figure 8, the net trend shown was down in both
number of species and number of organisms for Al, A2 and Al2.
All except A7 showed a sharp drop in April 1978.
At station A2 the net trend was down less sharply in
number of species and more sharply in abundance; the species
numbers at A2 appeared to be the only ones on the rise in
October 1978 of the five stations.
At A3 the species down-trend paralleled that of Al, but
the abundance trend was steeply upward, as was that of A7.
At A7 the species diversity net trend was upward but had
turned downward in October 1978. The number of organisms is
sharply up by the end of October 1978, but the oscillations
were extreme in the 1977-78 period.
It is important to look at the beginning and end points
for these trends. However, the events in waste treatment in
the harbor can be tracked because benthic worms, in particular,
reproduce year around and an area can be recolonized within
as little as two weeks to a month. Thus, A7 and A2 appeared
to increase in species numbers in June 1977 when TITP went on
-------
180
HE 6
secondary, while Al decreased in both species and numbers,
due perhaps to fewer nutrients. All but Al increased in numbers
in June. In September, both species and population numbers
dropped steeply at all four stations except A7, which had a
slight increase in numbers. One cannery hooked up to TITP in
October and the other in December 1977. The sharp drop in
species and numbers continued through January and April 1978
instead of showing the usual late winter increase, except at
A7, which showed a small increase in species and very large
population increase. Stations Al and Al2 showed small expected
January increases before the April lows.
In July 1978 all the stations in Figure 8 increased sharply
in species except A7, whose peak was blunted. All the stations
except A7 showed moderate (Al) to sharp increases in popula-
tion during the period of TITP release of high BOD and suspended
solids. In October, after stability of sorts returned, all
stations showed a drop in species except A2, while populations
were up at A3 and A7 (nearest TITP), but down at all other
stations.
Data for the other stations in the area, including those
in Figure 9, show a similar pattern; when canneries and/or TITP
were enriching, it appeared that most of the outer harbor
stations were enhanced and increased in diversity and popula-
tions. Stations A4, A8, and All showed net drops in numbers
of species, while A9 was about even in spite of the impact of
the Sansinena spill. In numbers, A4 decreased greatly, A8 was
increased slightly and All had a net increase. All showed
fluctuations but the September 1977-April 1978 period showed
the widest shifts. When TITP is on full secondary treatment,
it appears that A7, All and, at times, A2 will increase (become
enhanced) and the other stations decrease dramatically. When
canneries or primary TITP were on, A7, All and sometimes A3
retreated while the others increased.
The number of species at Al would probably always be
lower because of the freshwater effluent, sometimes chlorinated,
from TITP and storm runoff. Station A9 (Figure 1) is of interest
because it is across the main channel and in a different hydro-
graphic gyre; it was also the site of the Sansinena tanker
explosion and Bunker C spill, which was intensively studied
by Harbors Environmental Projects (Soule and Oguri, 1978).
Station A9 (Figure 7), which had been sampled a few days before
the explosion, showed a steady drop in species and numbers from
December 1976. By September 1977 species diversity had recovered
greatly and populations recovered to a lesser extent. However,
winter storms, high temperatures, and reconstruction of the pier
released much buried tar into the water and drove the numbers
sharply down through January to an unprecedented low in April
of 1978. These events can clearly be marked in the benthic
plot. By July 1978 counts and species had increased so that
the net slope for abundances for seven years was negligible
-------
HE 7
181
and the slope for species numbers was slightly up. The prom-
inent peak in numbers occurred in the fall of 1973, while the
highest peak in species occurred in the spring of 1976.
Qualitative Evaluation of Species Composition
At the representative stations selected for graphic com-
parison, notes were made on the most abundant species, as they
were first noted in the AHF (1976) report. Species were re-
corded as abundant if they constituted a large percentage (usu-
ally 35% or more) of the total animals. Common species usually
occurred in quantities of over 2,000/m2.
Station A1
Seasonality. Throughout the 1972-1978 sampling, the popu-
lations of the most numerous species, M. aaliforniensis and
Tharyx, show wide fluctuations, with peak size in spring months
and lows in the autumn. Other species tend to be more stable
but still show this seasonality at station Al.
General Trends. A marked reduction in population size for
all species began in October 1975 and continued to October 1978.
Diversity appeared to increase from October 1975 through March
1977; however, this was probably a result of multiple grab sam-
pling. For this period the counts of individuals were adjusted
to per-grab averages but there was no way to deal with the in-
crease of rarer species encountered. A prime example of this
occurred in December 1976 when 13 grab samples yielded 97 taxa.
In section IVB on multiple discriminant computer analysis only
the Polychaeta and Mollusca are used to aid in correction of
this factor.
Recently there has been some indication of faunistic change.
Beginning January 1978 Mediomastus aaliforniensis was no longer
numerically important. The October 1978 sampling stands out,
as both Tharyx and M. aaliforniensis were virtually gone. These
two had comprised 60% of the total harbor population in 197 3.
However, although ranking and species changed, the fauna was
otherwise typical of the outer harbor area (Table 2).
Station A3
Seasonality. Mediomastus aaliforniensis became dominant
in the fall-winter periods of 1972-74. No other cases of true
dominance occurred. Tharyx tended to be the most numerous an-
imal in the summer months.
General Trends. The number of individuals declined in the
non-summer periods beginning in 197 5 through the present sam-
pling. The multiple grab samples taken from June 19 75-March
19 7 7 artificially increased the total taxa figures for this time.
There have been no essential changes in the faunistic composition
-------
182
IIE 8
since the AHF, 1976 report (Table 3).
Station A7
No clear temporal patterns were seen in population size
or species diversity because of variability. Generally, when
population size was large, order-of-magnitude differences oc-
curred between dominant and second-ranking species. In the
23 sampling periods between March 1971 and October 1978 Arman-
dia b-Loaulata dominated twice and Cavitella capitata dominated
18 times. These two species and the nematodes are characteris-
tic of disturbed (variable) or polluted habitats (Reish, 1959)
(Table 4).
Numbers of molluscan and crustacean species began increas-
ing in 1978. In July 1978 species diversity was very high for
this station. Faunal composition of the October 1978 sampling
showed radical differences. The polychaete Mediomastus aali-
forniensis dominated, and the species found resembled outer
harbor or channel faunas.
Station A12
Seasonality. No strong seasonal patterns occurred, although
there was a tendency for population size to increase in spring-
summer periods (Table 5).
General Trends. The faunal composition has been quite
stable at this station, which was one of the richest in the
outer harbor. Changes in rank among the four abundant species
did happen, but only once, in August 197 2, when a single species
(Tharyx) was dominant.
There has been a decrease in the numbers of individuals col-
lected here since October 1975. Again, multiple grabs taken from
October 1975 through March 1977 for other purposes cloud the
diversity aspect, but at least permitted maintaining the long-
term baseline which would otherwise have been dropped.
Biotic Characteristics of Soft Bottom Habitats
Soft bottomed habitats with easily disturbed sediments favor
deposit-feeding organisms. These animals turn over the sediments
during feeding or burrowing, impeding to some extent the settle-
ment of suspension-feeding species (Levinton, 1973). However,
suspension feeders often cannot tolerate the turbidity levels
associated with unconsolidated bottoms. The activities of
deposit-feeding animals can turn over the entire bottom of an
area in a few years (Gordon, 1966). The outer harbor is an
excellent habitat for deposit-feeders.
Enrichment of a benthic community can produce marked
increases in the numbers and biomass of animals. In Scotland,
-------
HE 9
183
enrichment of a sea loch produced a maximum of up to 60,000
animals/m2 in 3 to 4 years (Raymont, 1950). This is remarkably
like the situation that occurred in Los Angeles Harbor in
1973-74 (Figure 10) following pollution abatement in 1970,
where numbers up to 80,000/m were encountered within the next
four years (AHF, 1976; Soule and Oguri, 1976). This contrasts
with the populations seen (Figure 11) after reduction in
nutrient wastes.
Tenore (1975, 1977) and Tenore and Gopalan (1974) found
that improvement in the nutritional quality of detritus by
enrichment was important in the growth of polychaete worms.
Proteinaceous wastes or the bacteria and protozoa supported
by finely ground sewage wastes served as a better food source
for the benthic fauna than fresh plant detritus.
Areas directly under outfalls often have depleted faunas.
This may be due to high organic content, toxic components,
chlorination turbidity, the impact of fresh water injected into
a marine environment, or to other unidentified factors. Depos-
it feeders in such areas often cannot turn over the sediment
swiftly enough to prevent decomposition in areas with high or-
ganic content (Nichols, 1974); however, no excessive buildup
has been seen in the Los Angeles Harbor outfall area. Pelecy-
pods increase in abundance with up to 3% organic content, but
decline at greater concentrations in the sediment due to decom-
position and anoxic conditions (Bader, 1954) . Concentrations
in the harbor are about 1.5%. Many fishes and invertebrates
cannot tolerate the chlorine residues in treated sewage (Emer-
son, 1976; Bellanca and Bailey, 1977), and toxic chloramines
or organics may be formed (Oliver and Carey, 1977). Fresh wa-
ter used to flush wastes and runoff can prevent settlement of
all but the most euryhaline species. Stone and Reish (1965)
documented the effects of seasonal rain runoff on recolonization
in the Los Angeles River. Such fluctuations are considered
healthy for the prevention of a few organisms from completely
dominating an estuary or bay.
Around the immediate vicinity of outfalls, species such as
Capitella aapitata are found, which are hardy, opportunistic,
fast-growing species that thrive in the absence of competition
(Grassle and Grassle, 1974). Although they have often been con-
sidered to be indicators of polluted conditions (Reish, 1959;
Word, Meyers, and Mearns, 1977), the species occurs in many un-
stable (variable) environments where rapid growth and short,
year-round reproductive cycles give them an advantage.
Beside varying spatially according to the amount of enrich-
ment, benthic animals in the harbors vary in size and quantity
according to the season. Like most shallow-water areas, these
bottoms receive varying amounts of sunlight, freshwater runoff,
and primary production by plants during each year. The repro-
ductive cycles of the invertebrates are attuned to these changes.
-------
184
HE 10
The year-class phenomenon, in which recruitment can vary enor-
mously from year to year, is prevalent (Grassle and Sanders,
1973). The combination of spatial and temporal patterns pro-
duces a shifting mosaic of benthic populations.
CONCLUSIONS
The benthic invertebrates found in outer Los Angeles-Long
Beach Harbors are species generally characteristic of soft bot-
tom habitats. Polychaete worms, gammarid amphipods (small
shrimp-like crustaceans), and small clams are common. Ghost
shrimp (genus Calliartassa) , sea pens, gaper clams (Tresus nut-
talli), and tube anemones (order Ceriantharia) are among the
larger animals in the area.
The general distribution patterns of the harbor have not
changed greatly in 1978 from the 1973-74 period (AHF, 1976).
There are still gradations from inner to outer harbor and from
the outfalls area to the outside of the breakwater.
The populations in the enhanced area (Soule and Oguri,
1976) numbered greater than 25 species and 35,000 individuals
per m2 in 1973-74.
The mean number of species/m2 rose four-fold from 1971
(14) to a high in 1976 (57). It has now dropped to pre-1973
levels (41) , (Figure 2; Table 1) .
The mean number of individuals/m2 rose from about nine-
fold between 1971 and 1973, the peak; the mean declined by 15%
of 1974 levels in 1975, by a severe 56% of 1975 levels in 1976,
and leveled off in 1977. There was another small drop in 1978,
placing the 1976-78 means at less than half the 1972 level.
Thus, both species numbers and population numbers have
dropped over the last three years, precisely the period when
so-called cleanup measures were instituted for cannery and sew-
age wastes. This cannot be blamed on rainfall, or lack of it,
because rainfall was low in the winter of 1972-73, yet counts
rose dramatically in 1973. The winter of 1975-76 had low rain-
fall, but populations declined.
A nearly four-fold decrease in benthic organisms between
1973 and 1978 represents a severe drop in the food supply of
obligate or facultative benthic feeders such as demersal fish
species and invertebrate predators. At station Al, declines
in the two major species Thavvx sp. and Mediomastus aal-Lfovni-
ensis (Capitata ambiseta) are notable since this would indicate
a decrease in enrichment at the perimeter of the harbor.
LITERATURE CITED : See Section VI.
-------
2
Table 1.	Annual Mean Numbers of Benthic Species and Organisms Per M .
(ispecies/#organism)
Stations
1971
1972
1973
1974
1975*
1976*
1977*
1978
Mean
A1
16/553
52/ 7,347
54/ 6,645
68/10,580
62/ 6,706
79/ 2,300
49/ 2,580
40/ 1,560
53/ 4,784
A2
24/ 4,117
52/29,284
66/35,360
64/38,296
62/18,133
65/17,060
74/10,993
46/ 4,412
57/19,707
A3
18/ 3,213
51/24,089
53/16,395
52/17,836
62/11,353
73/ 6,472
64/ 8,000
47/ 4,949
53/11,538
A4
4/ 1,345
19/17,169
6/ 8,331
7/ 2,728
17/ 9,910
23/ 1,932
50/15,363
30/ 9,128
20/ 8,238
A5
4/90
10/ 6,074
15/10,192
4/684
15/19,040



10/ 7,216
A6
5/77
4/178
10/12,272
6/ 2,332
29/54,064



11/13,785
A7
7/ 1,680
14/ 6,603
14/12,256
17/ 7,930
10/ 2,665
8/808
6/ 3,737
19/11,185
12/ 5,858
A8

39/30,362
57/55,093
58/21,000
54/ 3,600
t 71/ 5,640
48/ 1,813
42/ 8,244
53/17,965
A9
36/11,810
44/32,118
47/45,456
46/37,900
44/16,155
71/24,952
48/11,370
36/ 9,512
47/23,659
A10

44/19,984
48/53,520
47/28,616
51/36,464


48/21,851
48/32,087
All


64/21,880
54/26,040
48/13,961
68/ 5,220
62/ 6,408
32/ 5,701
55/11,316
A12

60/28,200
66/29,280
59/30,864
62/13,161
55/ 4,790
63/ 7,84 3
42/ 4,808
58/16,992
A13






69/15,970
48/ 5,068
59/10,519
A14






39/ 1,590
41/ 5,845
40/ 3,718
A15






49/ 6,450
39/14,836
44/10,64 3
A16






58/ 4,600
48/ 6,560
53/ 5,580
Al7






4 3/ 2,740
50/ 4,715
47/ 3,728
B8

50/ 9,94 0
61/41,403
62/34,012
62/15,728

69/16,290
46/ 3,888
58/20,210
B9

50/ 9,420
66/41,200
58/27,008
60/21,696


48/ 4,240
56/20,713
Mean
14/ 2,861
38/16,982
45/27,806
4 3/20,416
46/17,331
57/ 7,686
53/ 7,716
41/ 7,441
42/12,987
* multiple grabs increased diversity
oo
cn
-------
186	HE 12
Table 2. Benthic Species at Station Al, 1978.
Abundant Species
Polychaeta:	* Total
Indiv.
Msdiomastus aaliforniensis (formerly Capitita ambiseta) 21%
Tharyx sp. (Tharyx ?parvus)	9%
Prionospio (Apoprionospio) pygmaeus	4%
31%
Common Species
Polychaeta:
Prionospio steenstrupi (formerly P. nr.malmgveni)
Chaetozone setoea
Sigambra tentaaulata
Mollusca:
Parviluaina tenuiseulpta (Parviluaina sp.)
Tellina modesta
Mysella pedroana
Table 3. Benthic Species at Station A3, 1978.
Abundant Species
„ , ,	% of Total
Polychaeta:	mdxv.
Mediomastus aaliforniensis (formerly Capitita ambiseta) 32%
Tharyx sp. (tharyx Iparvus)	15%
Prionospio (Minuspio) airrifera	4%
Cossura Candida	3%
5T%
Common Species
Polychaeta:
Lumbrineris
Nephtys aornuta franaisaana
Nereis procera
Mollusca:
Tellina modesta
Maooma nasuta
Parviluaina tenuiseulpta {Parviluaina sp.)
Prototkaaa sp. juveniles
-------
HE 13
187
Table 4. Benthic Species at Station A7, 1978
Abundant Taxa
% of Total
Polychaeta:	Indiv.
Capitella oapitata	55%
Armandia biooulata	5%
Poly do pa ligni	2%
Nematoda:
Unidentified nematodes	13%
80%
Table 5. Benthic Species at Station A12, 1978.
Abundant Species	, Qf Total
Polychaeta:	Indiv"
ThaPyx sp. (ThaPyx Ipapvus)	37%
Cossupa aandida .	17%
Mediomastus oalifopniensis (formerly Capitita ambiaeta)	17%
Taubepia oaulata (formerly Papaonis gpaeilis oculata)	6%
77%
Common Species
Polychaeta:
Nephtys oopnuta fpaneiscana
Chaetozone oopona
Lumbpinepis
Mollusca:
Papviluoina tenuisculpta (Papviluoina sp.)
-------
B6J
WILMINGTON
LONG BEACH
B8&
C6*
D4*
3*
64.
D5*
All
B2*
Pier J;'
01*
D9*
B3*
B0.
D6-
A7
D7*
SAN
PEDRO?
BIO*
Lang Beach Harbor
B9*
A3*
A3
A17*
A2*
A2b'
,10*
B1*
A9*
Kill II Mills
A1*
Figure 1. Benthic Sampling Locations (TITP Study Area in Box)
AO*
Harbors Environmental Projects
University of Southern California
00
00
H
-------
I IE 15
189
mean if
¦peciss/r
-q mean $
individ/m
30,000..
25,000-.
-40
-30
10,000-
D-
5,000-
71
73
77
78
Figure 2. Annual means of benthic species and number of benthic
individuals/m2 for stations sampled, 1971-1978.
(multiple grabs enhanced diversity in 197 5-77)
-------
p90
100n
NUMBER OF SPECIES/M
-85
95-
NUMBER OF ORGANISMS/M
-80
90-
-75
85-
-70
£0-
75"
-60
70-
n
-55
2 65'
-50
60-
55-
M
U40
-35 ll £
m 45-
-30 E
35
30-
-15
25-
-10
20-
15-
Mar Jun Dec Jan Mar Aug Nov Mar Aug Nov Feb May Aug Nov Feb Oct Jan May Aug Dec Mar Jun Sep Jan Apr Jul Oct
1971	1972	1973	1974	1975	1976	1977	1978
FIGURE 3. NUMBER OF BENTHIC SPECIES AND ABUNDANCES AT STATION A1
OUTSIDE LOS ANGELES HARBOR, 1971-1978
-------
100-1
r90
95-
-85
NUMBER OF SPECIES/M2
NUMBER OF ORGANISMS/M*
90-
-80
85-
-75
80-
-70
75-
70-
-60
-55 Z
Z 65-
60-
-50
rn
CO
-45
55-
-40
-35 u.
-30
-25
35
-20
30-
25-
-15
-10
20-
15
-5
r
Mar Jun Dec Jan Mar Aug Nov Mar Aug Nov Feb May Aug Nov Feb Oct Jan May Aug Dec Mar Jun Sep Jan Apr Jul Oct
T
1977
1978
1971
1973
1974
1975
1976
1972
FIGURE 4. NUMBER OF BENTHIC SPECIES AND ABUNDANCES AT STATION A2
OUTSIDE LOS ANGELES HARBOR, 1971-1978
-------
100
95
90-
85
80
75
70
Z 65
lee
m
31 55-
O
"" 50
cn
^45
o
m 4o-
co
35k
30
25
20
15
NUMBER OF SPECIES/M
NUMBER OF ORGANISMS/M
1 I I I I I I I I I I I I I I 1 1 I I I I I 1 J I 1 I I
Mar Jun Dec Jan Mar Aug Nov Mar Aug Nov Feb May Aug Nov Feb Oct Jan May Aug Dec Mar Jun Sep Jan Apr Jul Oct
1971	1972	1973	1974	1975	1976	1977	1978
FIGURE 5. NUMBER OF BENTHIC SPECIES AND ABUNDANCES AT STATION A3
OUTER LOS ANGELES HARUOR, 1971-1978
-------
NUMBER OF SPECIES/M*
NUMBER OF ORGANISMS/M2
X
T—T
f-T
/ *
f \
\	
320 330 36i
I t
i i y
T
-35
-30
N
DC
Ul
H
UJ
5
"25 £
a.
o
O
•20 to
«E
CO
I H
O H
¦15 o £
u.
O
cc
Ui
¦1° S
z
r5
-1
Mar Jun Dec Jan Mar Aug Nov Mar Aug Nov Feb May Aug Nov Feb Oct Jan May Aug Dec Mar Jun Sep Jan Apr Jul Oct
1971	1972	1973	1974	1975	1976	1977	1978
KD
U)
FIGURE 6. NUMBER QF BENTHIC SPECIES AND ABUNDANCES AT STATION A7
OUTER LOS ANGELES HARBOR, 1971-1978
-------
100
r 90
NUMBER OF SPECIES/M
95-
85
NUMBER OF ORGANISMS/M2
90-
-80
85-
-75
80-
-70
75-
70-
-60
o
Z 65"
-55
60-
-50
m
55-
-40 £ H
O to
-35 u. °
S40-
-30 tr
35'
-25
30-
-20
-15
20-
-10
15-
Mar Jun Dec. Jan Mar Aug Nov Mar Aug Nov Feb May Aug Nov Feb Oct Jan May Aug Dec Mar Jun Sep Jan Apr Jul Oct
1971	1972	1973	1974	1975	1976	1977	1978
FIGURE 7. NUMBER OF BENTHIC SPECIES AND ABUNDANCES AT STATION A9
OUTSIDE LOS ANGELES HARBOR, 1971-1978
-------
I IE 21
100-
90-
80-
70-
60-
50-
I
,40-
30-
20-
10-
r

\
A?
/
N/
T
r-
/
195
i-SO
-45
¦40
1-35


-20
z
<

a

cc

o

u.

O
-15
cc

UI

m

2

3

Z
10
-5
MAR JUN SEP JAN APR JUL OCT MAR JUN SEP JAN APR JUL OCT
1977 i 1978	1977 ¦ 1978
FIGURE 8. COMPARISON OF Al, A2, A3, A7, AND A12, 1977-1978
-------
196
100
90
80
70H
A8
60
A4
a. 50-
A9
5 40-
30-
A11'
20-
10-
MAR JUN SEP JAN APR JUL OCT
1977 • I978
FIGURE 9. COMPARISON OF A4,
22
50
45
40
35
25 ^
20 <
A4
-15
Ad
-10
A11
A8-
A11
T
T
MAR JUN SEP JAN APR JUL OCT
I977 i I978
A 8 , A9 AND All, 1977-1978
-------
SCALE IN MllCS
WILMINGTON
LONG
BEACH

SAN
PEDRO
#	>30,000/m2
^	20,000-30,000
O	>10, 000-<20,000
o	>5,000-<10,000
•	<6,000
~	>40 speaies/m2
¦	>15-<40
°	
-------
SCALE IN MIlES
WILMINGTON
LONG
BEACH

SAN
PEDRO
on*1*
CABRILLO ::
BEACH
\
mwmmmjil!)
•	>20,000/m2
©	20,000-30,000
o	>10,000-<20,000
O	>5,000-<10,000
•	<5,000
•	<2,000
O	>40 speaies/m"
•	>15-<40
a	<15
10
00
H
M
to
Figure 11. Denthic Species and Organisms/M^, 1978.
-------
I IF
199
FISH EGG AND LARVAE SURVEYS
INTRODUCTION
The Los Angeles-Long Beach Harbor complex was shown to be
rich in fish species and numbers in 1972-1974 (Stephens, 1974;
AHF, 1976) and to be an important nursery grounds for larval
fish (AHF, 1976). The latter study emphasizing the anchovy
population sampled eggs and larvae from February 19 73 through
September 1974 at forty stations in San Pedro Bay.
In an effort to update information about larval fish pop-
ulation following initiation of secondary treatment in 1977 of
cannery and domestic sewage discharge into the harbor, new
studies were initiated in 19 78 for the City of Los Angeles Ter-
minal Island Treatment Plant.
During the initial study, anchovy (Engvaulis movdax) eggs
and larvae were sampled by means of horizontal tows at 4 m
depth. Since the initial study, the spawning stock of ancho-
vies in the harbor has decreased by two orders of magnitude
(Frey, personal communication); consequently the sampling pro-
gram was altered to effect capture of larvae from all parts of
the water column and to produce data comparable with egg and
larvae surveys conducted by National Marine Fisheries Service
(NOAA) (Kramer et al. 1972).
Understanding of spawning behavior, seasonal periodicity,
environmental factors and species interactions of commercially
important fish taxa have been investigated by a variety of gov-
ernmental and private concerns. Recently the emphasis has been
expanded to consider under-utilized or non-utilizable fish re-
sources .
The scope of this study encompassed factors that affect
larval population and adult population dynamics.
Concurrent with this investigation a monthly monitoring
program characterizing the abiotic and microbiotic parameters
of the harbor was conducted by Harbors Environmental Projects;
concurrently, trawls to assess adult fish populations were
being conducted by Stephens (Section IIA). Data generated by
those studies are presented to augment understanding of some
of the factors affecting larval fish populations. As knowledge
of all phases of the harbor ecosystem is increased, there is a
concomitant increase in the capacity to assess and predict the
effects of fluctuations or perturbations.
It appears from the studies conducted over the last five
years that adult populations of the numerically dominant species
(Genuonemus lineatus and Engvaulis movdax) have been declining.
The present study has attemped to provide information from ich-
thyoplankton and other data that will assist in exploring the
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IIF 2
possible causes of this decrease.
MATERIALS AND METHODS
Ichthyoplankton populations were sampled in the Los An-
geles-Long Beach Harbor area at ten stations (Figure 1) over
a period from January through November 1978. The earlier
study (AHF, 1976} sampled these ten stations as well as an
additional thirty, in the same general area.
In the original ichthyoplankton study, a 0.5 m plankton
net (333)i mesh) was towed at 4 m depth from a small boat at
low speed. In the current study, oblique tows using 333yi mesh
bongo nets were taken from the R.V. Golden Wesv (Figure 2),
such that while the boat was in motion, the nets descended to-
wards the bottom as cable was released and ascended to the sur-
face while cable was retrieved. From April through November
19 78, a second sampling method was added, a 0.5 m plankton net
(333ji), towed for five minutes from the R.V. Golden West near
the surface (< lm). The cod ends of both nets in the current
study were provided with screens of mesh size similar to that
of the nets. Bongo nets were equipped with a rocket-shaped
flow meter (General Oceanics); the 0.5 m ring net was equipped
with a four-blade, four-dial flow meter (Rigosha).
Table 1 summarizes the monthly sampling regime at each
station. During the first three months (January-March) bongo
tows were conducted at night. To facilitate comparison of da-
ta with earlier studies employing different sampling methods,
additional 0.5 m net tows were taken during the day for the re-
mainder of the study.
Samples were fixed on shipboard with buffered formalin and
returned to the laboratory for identification. Eggs were sort-
ed and counted, and engraulid eggs were further separated and
tabulated. Larvae were sorted and grouped by general charac-
ters, and then identified to the lowest possible taxonomic lev-
el (family, genus or species), except for unidentifiable, "un-
known" specimens. Literature references concerning early life
histories of marine fishes, as well as an established reference
collection, provided a source of comparison for identification.
A reference collection consisting of larval specimens from the
current study was established and identifications were veri-
fied by personnel of the National Marine Fisheries Service, La
Jolla, California. Counts were tabulated and a conversion fac-
tor based on flow meter data was used to quantify number of
larvae or eggs in #/m3 of water sampled.
Concurrent with this study, the Harbors Environmental
Project was sampling forty stations monthly in Los Angeles-
Long Beach Harbors. Data gathered on dates and at stations
correspondingly close to trawl dates and ichthyoplankton sta-
tions (Al, A2, A4, A7, B9, B10, Bll, C3) were assimilated into
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201
composite monthly averages to provide necessary background
for interpretation of ichthyoplankton data.
Surface water was collected by bucket at each station,
and samples were taken for nutrient analysis (nitrate, nitrite,
ammonia and phosphate levels) in the laboratory. Samples were
filtered on board for spectrophotometric chlorophyll a anal-
ysis in the laboratory. Nutrient and chlorophyll analyses
were performed according to Strickland and Parsons (1972);
Section I, this volume. Temperature and salinity data were
taken with a Mark V Martek Corp. remote probe unit.
RESULTS
Abiotic Data and Phytoplankton
Surface temperatures in the harbor (Figure 3) fell from
a year-end high in 1977 (17.3°) to a winter low of 15.4° in
January 1978, rose through the spring, and fell off in early
summer in July. Temperature resumed an upward trend toward a
maximum in October (20.4°) and then fell rapidly through No-
vember and into December. Salinity values fluctuated for the
first four months of the year, leveled off in March through
October and then began to increase toward a seasonal high la-
ter in the year.
Figure 4 shows values for two of the primary nitrogen
sources available to photosynthetic algae and bacteria. Ni-
trite (NO2) levels remained fairly constant (.11-. 27 jig-at
N/1) throughout the year except for a protracted peak between
July and September. Nitrate (NO3) levels increased through
the early part of the year and then fell abruptly to a low
summer plateau, increasing in the fall. Figure 5 shows the
mean values of ammonia nitrogen (as NH4) and phosphate (PO4)
phosphorus. Ammonia values follow those of the nitrate
sources, while phosphorus levels remain fairly stable.
The seasonal fluctuation in harbor chlorophyll a levels,
a measure of plankton biomass, is shown in Figure 6. Values
are low during the early part of the year (1-2 mg Chi a/m3)
increased to a high in September and showed another maximum in
November (5 mg Chi a/m3).
Ichthyoplankton
Displayed in Figure 7 are the seasonal variations in egg
and larval stocks in the harbor area for 1978 and for the orig-
inal study in 1974, expressed as mean number/100m3 of water
filtered. The largest incidence of eggs and larvae for both
studies occurred in the early part of the year (January-March).
1974 data are taken from Figure 7.6 of the orginal study, which
represented numbers of all eggs and larvae other than from
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IIF 4
anchovies. (Information elsewhere in the text showed engraulid
data to be significant only in the first three months, where
they provided a modest increase, e.g. total larvae including
anchovies peaked at 10 3/100m3 in February.) While the patterns
of seasonal variations are similar for both 1978 and 1974, con-
centrations are greater by orders of magnitude in 1978 than in
1974. Differences in collection depth would have influenced
this increase to some extent, as discussed in a later section.
While abundance of larvae is at a maximum in the early
part of the year, Figure 8 illustrates that the number of spe-
cies of fish spawning has two peaks; one in the early part of
the year and then one in fall.
Over the course of the year the abundance and composition
of the ichthyoplankton population varied greatly. In the ini-
tial part of the year larger numbers of sciaenid larvae
(white croaker, Genyonemus lineatus) dominated larval counts;
the majority of the correspondingly high numbers of eggs were
also probably from sciaenids. Table 2 represents a taxonomic
breakdown of eggs and larvae encountered in this study, iden-
tified to the lowest possible taxonomic level or, occasionally,
a most likely choice when absolute identification could not be
made {e.g. Paraliehthys/Xystrenrys).
Table 3 lists larvae identified in Table 2 in descending
order of abundance, both as total numbers of larvae captured
and as number/100m3/yr. Also listed for each larval form are
the numbers of samples (62 total) at which each occurred as
well as the number of months (out of a possible 9) in which
each was seen. The sciaenid species Genyonemus lineatus and
engraulid larvae constitute the two most abundant larvae en-
countered, while gobiid and engraulid larvae showed the highest
numbers of occurrences.
Species Distributions
Figures 9 and 10 show the distributional pattern of eggs
and larvae throughout the harbor. Station 4, nearest the can-
nery outfall area and TITP discharge areas showed high numbers
of eggs and larvae. Station 2, near the entrance of the har-
bor also showed high numbers of eggs.
Figures 11, 12, 13 and 14 represent stations which showed
the highest numbers of larvae captured for the four top-ranking
larvae encountered. The majority of Engraulidae larvae were
still found outside the breakwater (Figure 11), as was true
in the earlier surveys (AHF, 19 76). Water movement into the
harbor through the gates probably carried large numbers of lar-
vae into the station near the entrance. Abundance decreased
as the larvae are carried further and further into the harbor.
The lowest abundances were found in the outfall area and in
the Long Beach Back Channel as in the earlier studies.
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203
Figure 12 illustrates the pattern of larval abundance of
the Sciaenidae, principally Genyonemus. As in the original
study, sciaenids showed large concentrations in the Back Chan-
nel (St. 17) and around the outfall area. Environmental
Quality Analysts/Marine Biological Consultants (1978) reported
that Genyonemus adults comprised 49% of other trawl collections
from Long Beach harbor, where the majority of stations were in
the Back Channel.
The Gobiidae data show (Figure 13) that most of the
larvae stay near the areas of rocky habitats preferred by the
adults. As in the last study, the most productive stations
were those in the channels and near the breakwater. Figure
14 for the Blenniidae also shows that station 17 was an area of
high larval abundance, as well as station 5. The blenny lar-
vae were found near the environments of the more stationary
adults, as were the gobies. But unlike -the gobies, blennies
did not show significant numbers of larvae around station 16.
Both families have eggs that attach to the habitats, so they
would not contribute to the eggs found in the survey.
Table 4 compares the larvae from plankton tows and the
adult fish taken by the trawls for both 1974 and 1978. Each
is ranked by abundance of total numbers captured. Of the most
abundant larvae captured for 1978, only the midwater myctophid,
Stenobraahius, is unexpected, though it has been shown to come
inshore to spawn. Two of the ten most abundant adult species,
Phanerodon and Hyperpvosopon, are live bearers (embiotocids)
with larvae well developed at birth; these would show a high in-
cidence of net avoidance. No representatives of gobies or
blennies were found in Stephens' adult fish trawls among the
ten most abundant species, while both figured predominantly in
larval tows; however, Environmental Quality Analysts/Marine
Biological Consultants (1978) data showed that Lepidogob-ius
ranked third in abundance prior to thermal input, decreasing
in abundance to a rank of tenth afterward.
Clinids and the pomacentrid Chvomis showed high numbers
of larvae in 1974, and fewer in 1978. Some of the explanation
lies in the restricted area of the 1978 tows. The tows for
the present study were confined to the harbor proper and im-
mediately outside. Clinids and pomacentrids in 1974 showed
larval abundances outside the harbor generally, concentrating
in an area outside the scope of this study. The only larvae
which were dominant in 1974 inside the harbor and which were
not dominant in 19 78 were representatives of the Cottidae.
Members of this family, commonly called sculpins, do occur in
the harbor. Several genera, Clinoaottus, Leptoaottus and
Saorpaeniahthys, occur at numerous locations around rocky hab-
itats such as the breakwaters and rocky shores of Terminal Is-
land. A reason for their decline in larval abundance cannot
be conjectured because of lack of sufficient information about
the population dynamics and habits of the adults.
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I IF 6
Seasonality
The highest rate of spawning activity, which occurred in
January, February and March, coincided with seasonally high
levels in chlorophyll a concentrations. During this period
two families, the Sciaenidae (Genyonemus) and Engraulidae
(probably Engraulis), accounted for 93% of the larvae captured
(62% and 31%, respectively). Since the pattern of egg spawn-
ing should correspond with the larvae in the area, it can be
assumed that the preponderance of eggs were also sciaenid.
Abundance figures in 1978 are 20 times those in the pre-
vious study for the first three months (Figure 7) although
this is biased by the collecting methods and is not quantifi-
able with accuracy. A decrease in larval numbers followed
during the spring and summer, with an increase from September
through November, corresponds with the maximum numbers of
about 20 species of fish larvae collected in February and Sep-
tember (Figure 8). The larvae and eggs develop faster in the
summer with the increased temperature and therefore settle
faster. Coupled with reduced numbers of adults spawning, the
abundance of eggs and larvae would be expected to be lower in
summer.
Larval fish reached the lowest levels in August. In the
present study, August was the first month in which surface
samples were analyzed quantitatively. The decrease shown was
probably due to differences between bongo and surface tows.
Abundance returned to June levels in September and November
where surface tows also were taken. Comparison of data for
simultaneous surface tows and bongo tows showed that the for-
mer method underestimated abundance by a factor of 10 as com-
pared to bongo net totals.
The species using the harbor area as a spawning ground
showed a periodicity paralleling phytoplankton peak biomass
as noted previously in Figures 7 and 8. A spring bloom and
fall bloom put large concentrations of cells in the water which
can possibly be utilized by larvae. Some of the species showed
spawning throughout the year (Hyp soblennius and engraulids).
Others showed spawning restricted to the winter/spring period
of January through April {e.g., Genyonemus and Stenobrachius),
or the fall period of August through November (Peprilus, Sym-
phurus, and Gxyjulis).
DISCUSSION
The Los Angeles-Long Beach harbor system is a complex
ecosystem, adjusting to influences of natural processes and
accommodating discharge of urban waste and sewage effluents
from commercial and industrial concerns. The effects of a
quiet water harbor environment and input of effluents have
combined to produce a near-eutrophic system with high
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205
production estimates at all levels.
Attempts have been made by the City of Los Angeles and
the Regional Water Quality Control Board to improve water
quality throughout the harbor system over the past five years,
in compliance with federal legislation. Part of the "clean-
up" activities focused on the fish cannery facilities and the
discharge of processing wastes into the harbor. In 1975 the
canneries began to put discharge through dissolved air flota-
tion (DAF) systems to reduce the suspended solids released to
the harbor. In the spring of 1977, the Terminal Island Treat-
ment Plant initiated secondary treatment of harbor industrial
and sewage wastes. In October 1977, one of the two effluents
from the canneries was connected to the TITP facility and by
January 19 78 the second was connected, subjecting the process
water to secondary treatment as well.
In order to assess changes in water conditions it is nec-
essary to compare data from years prior to implementation of
treatment with data from subsequent years. Harbors Environ-
mental Projects have collected data since 1971, and published
in particular on conditions in the harbor for 1973-74, (AHF,
1974). Included in that volume were data concerning both
abiotic and biotic factors, both of which have been considered
in the scope of the present study. Other physical and biolog-
ical data were published in the journal Marine Studies of San
Pedro Bay3 California (Soule and Oguri, eds.).
Abiotic Factors
Temperature and salinity values (Figure 3) over the course
of the year reflect seasonal patterns seen up and down the Cal-
ifornia coast; i.e. water characteristics for the harbor are
mediated by water mass factors affecting the entire coast,
primarily, variability in the intrusion of the Davidson Current
(Oguri, 1974).
In the harbor in the initial part of the year, temperature
values approached a mid-winter minimum (15.4 C) and rose
through the spring and summer to a fall maximum in October
(20.4 C). The 5 degree C change for the entire year reflects
the warm water trend the Southern California Bight has been
experiencing for the last several years; the minimum was about
4 degrees higher than would normally be expected.
Salinity values in the harbor fluctuated between 37 ppt
and 24 ppt during the early part of 1978. This wide oscilla-
tion resulted from sampling surface waters during or immedi-
ately after rains that coincided with sampling dates. As the
rainy season tapered off, the surface salinity values stabil-
ized at approximately 30 ppt and began to increase toward a
winter maximum in December. The values found in 1978 were well
within the range of values found during 197 3 and 1974.
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I IF 8
Nutrient values for nitrate nitrogen (NO3), nitrite nitro-
gen (N02), ammonia nitrogen (NH4) and phosphate phosphorus
(PO4) are illustrated in Figures 4 and 5. Patterns of maxima
and minima for coastal areas have been well documented
(Raymont, 1963). The patterns for the harbor were similar,
with peaks in values in spring and winter and minima in the
summer and fall months.
Nitrate values rise steadily to a spring maximum in March
and drop rapidly to minimal values in May. The spring maximum
in the harbor coincided with nutrient turnover and increase in
NO3 values found in coastal waters in the early months of the
year (Thomas, 1974); flood control runoff entering the harbor
during the winter rainy months may also contribute to this peak
(AHF, 1976). Nitrite values for the harbor showed a character-
istically narrow range, as well as low levels for the entire
year, consistent with low levels in the bight region (with the
exception of an anomalous peak in August coincident with the
TITP malfunction). Ammonia and phosphorus also show typical
patterns with the exception of unexpected peaks in midsummer.
These peaks also corresponded in time with the breakdown of
the TITP plant; the effects of this temporary cessation in sec-
ondary treatment will be discussed in a later section.
Text Table 1 compares nutrient ranges for 1974 and 1978.
Generally, the nitrogen sources appear to have lower concentra-
tions in 1978 and phosphate levels are slightly higher, but the
differences are not large. Considering that in this system
medians of ranges are likely to be higher than means using all
pertinent data, it appears that both years 1974 and 1978 are
similar in nutrient characteristics.
Text Table 1,
Nutrient Levels (ug-at/1) in Los Angeles-
Long Beach Harbors in 1974 and 1978.
Nutrient
1974*
Range Mean
1978
Range
Mean-]_ Mean 2

N03
m2
nh4
P04
1.52-13.86
0.12-0.81
0.41-11.9
0.46-1.75
7.35
.31
4.40
1.10
0.9-14.2 (69.0)**	5.10	5.37
0.11-0.27 (3.2)	0.17	0.22
1.2-7.8 (60.6)	3.70	4.25
0.7-2.4 (6.7)	1.55	1.53
* data from AHF (1976)
** anomalous values from breakdown of TITP plant (in paren-
theses) are excluded from range of characteristic nutri-
ent levels for 1978
*** mean2 calculated with anomalous values included
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207
Phytoplankton Factors
Nutrient concentrations, temperature and salinity affect
adult fish populations primarily indirectly. These factors
as well as others (e.g., light levels and silicate concentra-
tions) directly mediate phytoplankton biomass and species
succession (Raymont, 1963; Ryther and Dunstan, 1971; Droop,
1973; Rhee, 1974; Tilman, 1977). Phytoplankton biomass, which
can be estimated by chlorophyll a concentrations, is closely
related to peaks in nutrient values, such that large drops in
nutrient levels reflect utilization of nutrients by phytoplank-
ton. Normal high nutrient levels and low chlorophyll a values
early in the year, during colder water periods, reflect season-
al nutrient regeneration and turnover, while phytoplankton
numbers may be low due to low levels of available light. As
day cycles lengthen, utilization increases, causing decreases
in nutrient concentrations, In summer the values remain low
due to utilization. In 19 78, only in May did nitrate and
phosphate approach levels reported to be limiting in culture
(< 2 yig-at P/l and <1.5 pg-at N/1) (Parsons and Takahashi,
1974). While some slowing of growth has occurred, it does not
appear that nutrient levels were a major factor in limiting
phytoplankton growth except for short periods during the year;
this parameter of primary production does not appear to have
changed significantly from 1974-1978.
Some evidence of the effects of a breakdown in the TITP
plant in late June to August 1978, appears in nutrient and
chlorophyll data. Ammonia and phosphate showed anomalously
high levels in July, and nitrites also showed a possibly rela-
ted peak in August, reflecting concentrations directly at the
outfall station (A7) or nearby; these levels dropped immediate-
ly to previous levels (Figures 5 and 6). Chlorophyll a biomass,
however, at stations near the outfall (A7 and A4), decreased
in July, suggesting a toxic effect on the phytoplankton, pre-
sumably from the wastes and control substances used. Data for
the harbor as a whole suggest that chlorophyll biomass has re-
mained relatively stable in 1977 and 1978.
Chlorophyll a values typically show a spring bloom in re-
sponse to the above factors, but data for 1978 did not show
this. One possible explanation is that phytoplankton blooms
can sometimes be extremely short in duration, as zooplankton
grazing can reduce phytoplankton stocks by orders of magnitude
in a few days (Fleming, 1939) . It is therefore possible to
miss a peak entirely when sampling only once a month. Nitrogen
concentrations decreased rapidly during the spring as if utili-
zation were in fact occurring. Therefore, it is possible that
the usual spring bloom occurred as in 1976 and 1977, but was
underestimated due to the sampling program.
Uptake rates were rather low in 1978, therefore assimila-
tion ratios are low as well. This may indicate a low C:N ratio
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IIF 10
in the harbor in 1978, perhaps as a result of a change in
carbonate equilibrium.
In comparing chlorophyll values for 1976-78 from Harbors
Environmental Projects data (this volume, section IIC), total
chlorophyll a biomass values were quite similar for 1977 and
1978 (35 i 1 rag Chi a/m^/yr). 1976 values were higher
(40 mg Chi a/nw/yr)' not markedly. The seasonal pattern
is essentially the same for all three years. Further, the
seasonal pattern inside the harbor parallels that seen at
station A1 outside the harbor proper, indicating, as stated in
section IIIC of this volume, that causative conditions are not
unique to the harbor, and implicating considerable seasonal
influence from the basic character of waters resident along the
coast. The effects of natural yearly oscillations cannot be
assessed from this short-term period; nevertheless, all indi-
cators except carbon uptake indicate that chlorophyll biomass
in the harbor has not changed significantly over the three
year period. This suggests that secondary treatment of can-
nery effluents has had little overall effect, and it is sur-
prising that the institution of secondary treatment of harbor
wastes begun in the spring of 1977 has not had a greater effect.
Aside from considerations of phytoplankton species composition
and annual fluctuations in water mass characteristics, the to-
tal phytoplankton food supply for larval fishes in the harbor
has not apparently become a limiting factor.
Fish Populations
Adult populations of fish in the harbor have shown a de-
creasing trend since 1974. Two important species, Genyonemus
lineatus (white croaker) and Engraulis mordax (Northern ancho-
vy) , have shown the most dramatic drops and thus heavily influ-
ence fish trawl data (Stephens, 1974; Section IIA, this volume).
For the numerically dominant fish in the harbor, Genyonemus,
a major factor in its decline is the removal of particulate
cannery wastes. In 1974, trawls in and about the cannery out-
fall area captured > 700 fish/trawl, with a preponderance of
Genyonemus. During the summer of 1973 Stephens captured more
than 25,000 juveniles in the outfall area. After introduction
of DAF treatment to coagulate wastes, fish populations around
the outfall dropped to around 160 fish/trawl in 1975 and 88
fish/trawl in 1976. In 1978 around the outfall area, the num-
ber of fish per trawl was 125, increasing in July to 993 fish/
trawl, exceeding 1974 values. This increase probably resulted
from the breakdown in the TITP plant, releasing sewage into
the area and causing a rise in BOD and suspended solids of 10
and 30 times, respectively. This evidence, with trawl data be-
fore and after DAF installation, as well as the report by Reish
and Ware (1976) on the feeding habits of Genyonemus, show that,
at least in part, the decrease in adult fish populations can
be attributed to a reduction in available food resources. The
DAF treatment removed available organic nutrients that were
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209
previously discharged into the harbor by the cannery (Section
HE, this volume) and probably had a far greater impact than
secondary waste treatment. Genyonemus is an omnivore (Reish
and Ware, 1976) and consumes large numbers of benthic poly-
chaete worms. A drop in organic nutrients and increased preda-
tion pressure on polychaete populations, affecting both abun-
dance and species composition, would greatly reduce the nutri-
ent source available to bottom-feeding fish.
The Northern anchovy population (Engraulis mordax) has
been declining in the Southern California Bight area for the
past five years. Acoustical surveys show a 1973 peak of 1.8
-2.0 x 10® T, less than 5 x 10^ t in 1978 and 3 x 105 t in
1979 (Frey, personal communication), showing a six-fold de-
crease in five years. In the harbor, though, based upon the
data of Stephens et al. (1974) and Harbors Environmental Proj-
ects (this volume, Section IIA), anchovy biomass has been re-
duced a thousand-fold. Possibly the installation of DAF treat-
ment by the fish canneries has also influenced the decline of
Engraulis in the harbor. The harbor has not had a dinoflag-
ellate bloom in recent years (Morey-Gaines, personal communica-
tions) which constitutes a large part of the larval anchovy's
diet (Lasker, 1975). The reasons for this are not clear but
may relate to temperature, or to bacterial populations de-
creases (Oguri, Soule, Juge and Abbott, Red Tide Conference,
1975). Bacterial populations dropped 30-fold following TITP
conversion to secondary treatment (Section III, this volume),
but there are no comparable data for previous years.
While adult populations of Genyonemus and Engraulis have
been greatly reduced, other species have more nearly maintained
their population levels since 1975. This suggests both that
waste treatment can have species-specific effects and that in-
dependent factors can operate in a complex system that may con-
fuse interpretation of cause-effect relationships.
Larval Abundance
Based on large numbers of juveniles encountered by Stephens
(1974) in the summer of 1973 it is reasonable to assume that
large numbers of eggs and larvae were spawned in the preceding
couple of years. However, the previous ichthyoplankton survey
did not show the expected high numbers of larvae or eggs during
the early months of 1973 and 1974 when Genyonemus spawns
(AHF, 1976). In contrast, in January and February, 1978, this
study found numbers of eggs and larvae that peaked at values
in the hundreds per cubic meter, which are extraordinarily
high (Figure 7) and indicate a highly productive area. In com-
parison, values for a recreational harbor in Santa Monica Bay
(King Harbor) showed larval densities of 44/m^ at the richest
station (McGowen, 1978).
A possible factor contributing to this abundance is that
several genera of the sciaenids {Bairdiella, Umbrina and
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IIF 12
Seriphue) exhibit cannibalistic or predatory behavior on eggs
and larvae (Maxwell, 1975); the flatfish Cithariehthys has
been known to have fish eggs in its guts (Reish and Ware, 1976).
A considerable decrease in the adult fish population between
1973 and 1978 could affect predation levels and thus would in-
crease the number of larvae and eggs in the water column.
Another factor that must be considered is the sampling
method. The original survey consisted in horizontal tows at
approximately 4 m below the surface. In the harbor, a chloro-
phyll maximum layer was found around 3 m (Section IIIC, this
volume). Lasker's (1975) work with anchovy larvae demonstra-
ted larval and egg stratification in and above the chlorophyll
maximum. He found that larvae without pigment in their eyes
(therefore nonfunctional) were found in high abundance above
the chlorophyll maximum layer, while sighted larvae were found
predominantly in the chlorophyll maximum layer, and abundance
above the layer exceeded that below by a factor of ten. Eggs
were stratified also, with a high abundance in and above the
layer. Furthermore, anchovy larvae with pigmented eyes show
significant net avoidance (P. Smith, 1972). Other larvae could
stratify in a similar manner. Over 60% of sciaenid larvae
from the oblique bongo tows, sampling the whole of the water
column, were without eye pigmentation, indicating successful
sampling of upper layers. While no evidence concerning pigmen-
tation is available from the earlier study, it is likely that
sampling at 4 m would result in a significant decrease in the
number of eggs and larvae captured relative to the actual pop-
ulation. Although the magnitude of this decrease cannot be
accurately estimated it is probably not sufficiently great to
account for the discrepancy in abundances between 1974 and 1978.
Possibly a ten-fold higher abundance in 1978 is a more reason-
able estimate.
The number of species 'of fish larvae collected each month
in 1978 ranged from 5-18 with a baseline around eight and two
peak periods in spring and fall (x = 11.6). Data for adult
fish species in the same area show a range of 0-15 and mean
around 7 for 1974-1978 (Environmental Quality Analysts/Marine
Biological Consultants, 1978), and in another study (Stephens,
1974), a range of 1-11 and a mean of 6. This higher number of
species in the ichthyoplankton collected in 1978 alone may re-
flect a higher diversity than in earlier years, although pos-
sibly the difference in sampling methods could allow a differ-
ent representation of species in the catches.
It is tempting to speculate that these high abundances of
1978 eggs and larvae will result in increases in future adult
populations, but further investigation is needed in order to
predict the fate of the high number of eggs and larvae (Hempel,
1965). Without additional sampling employing collection of
settled larvae, there are no direct means of determining suc-
cess of recruitment. It has been demonstrated (Lasker, 1975;
O'Connell and Raymond, 19 70) that first-feeding anchovy larvae
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I IF 13
211
require high concentrations of food particles of a certain
size. Cell size and abundance cannot be extrapolated direct-
ly from chlorophyll concentrations alone. Cell size distribu-
tion depends on the presence of suitable species and on their
succession in the phytoplankton. Limited information concern-
ing species composition and size distributions within the
phytoplankton is available (Section IIIC, this volume). Gen-
erally, red tide blooms were lacking in 1978, and without high
concentrations of dinoflagellates recruitment of anchovy larvae
cannot be expected to have a high rate of success. The data
are insufficient to predict future adult population abundances.
CONCLUSIONS
Secondary treatment has not significantly altered nutrient
conditions, which seem to follow seasonal trends that occur
throughout the Southern California Bight. This change also
has not significantly altered phytoplankton biomass, which
follows the-patterns of abiotic factors (nutrients, tempera-
ture, salinity and light levels) that are influenced by off-
shore events. Nevertheless, adult populations of fish in the
harbor have been declining since dissolved air flotation (DAF)
treatment was initiated in 19 75. Previous studies show that
the dominant adult fish species Genuonemus utilized the outfall
area for foraging on suspended cannery wastes, and also fed on
numerous benthic worms in the enhanced area of the outer har-
bor. The number of adult fish captured per trawl increased by
an order of magnitude for a brief period in 1978 coincident
with a TITP malfunction, when particulate sewage and industrial
wastes were discharged into the harbor at the outfall. The
fish numbers then declined after full recovery of the treat-
ment plant. High nutrient values occurred during the breakdown,
but chlorophyll biomass was slightly reduced. Chlorination
was also going on at that time.
The present study found higher counts of eggs and larvae
than were found in an earlier study; this is probably due to
reduced predation by adult fish and to more efficient sampling
methods. Larval species demonstrated discrete distributions
within the harbor according to habitat and food resource. Suc-
cess of recruitment of larvae cannot be predicted on the basis
of egg and larvae census alone, as was demonstrated by the
huge numbers of anchovy larvae offshore in 1975 which failed
to recruit for unknown reasons. The result has been a 4-fold
drop in spawning biomass off California by 1979. Therefore,
the contribution of these larvae to future adult populations
remains in the realm of speculation.
LITERATURE CITED See Section VI
-------
212
IIF 14
SCAll IH MIIIS
WllMlNGTON
17
16,
*5
*4
*7
*6
*15
Figure 1. Ichthyoplankton trawl stations, 19 78. Station 21
is approximately 2 miles off the breakwater.
////////
Figure 2. Diagram of sampling method using paired bongo nets.
Dashed line represents complete path towed through
sampling location.
-------
IIF 15	213
S o/oo T (°C)
38
00
36-
-21
34-
-20
32
19
30
18
28-
-17
26-
16
24-
-15
22
T
T
djfmamjj asond
Figure 3. Mean monthly temperature and salinity values
recorded in 1978. Monthly averages represent data
collected at selected harbor stations on sampling dates
most closely correlated to ichthyoplankton trawl dates,.
NO
12-
1.0
10
8--0.8
4--
2--
T
T
T
DJF MAMJJAS OND
Figure 4. Mean monthly NO3 and NO2 nitrogen levels in
jig-at/1 for 1978. Monthly averages represent data
collected at selected harbor stations on sampling dates
most closely correlated to ichthyoplankton trawl dates.
-------
214
I IF 16
NH4 PO4
2.0
1.5
5--1.0
Figure 5. Mean monthly ammonia nitrogen levels (o) and
PO4 phosphate levels (•) in pg-at/1 for 1978, Monthly
averages represent data collected at selected harbor
stations on sampling dates most closely correlated to
ichthyoplankton trawl dates.
5-
^ A-
3-
U
Cn
£
2-
0 J	1	1	,	1	J	1	1	1	1 I I
jfmamjjason
Figure 6. Mean monthly chlorophyll a levels in
mg/m^ for 197 8. Monthly averages represent data
collected at selected harbor stations on sampling
dates most closely correlated to ichthyoplankton
trawl dates.
-------
IIF 17
215
EGGS
1978 (total eggs)
1974 (total minus
anchovy eggs)
10,000-
1000-
ro
M
OJ
a
100
u
o
LARVAE
10,000"
1978 (total larvae)
c
c
d)
s
1974 (total minus
anchovy larvae)
1000-
100-
10
J
F
M
A
M
J
J
0
A
S
N
Figure 7. Mean number of fish eggs and larvae collected
per irw of water filtered for Los Angeles Harbor and
in 1974 from San Pedro Bay. Latter values represent
total numbers excepting anchovy eggs and larvae.
-------
216
I IF 18
20 -
% 15
•H
10
A
S
0
D
J
A
M
J
J
F
M
Figure 8. Number of species of fish larvae collected
during 1978.
-------
JCALt IN Mil*»
WILMINGTON
L,ONG
• EACH
SAN
PEDRO
CAMIILO
¦Each
MIAN HANCOCK fOUNDAIION
O 0-15
» 100-200
% 200-300
9 400-500
	 HAK1QB iNVtSQNMfHTAI HOJtCTS
Figure 9. Distribution of total number of larvae/m3 in the Los
Angeles-Long Beach Harbors in 1978.
-------
ICAU IN Mllti
WILMINGTON
IONO
¦ EACH
SAN
PEDRO
• 10-100
100-300
CABftmo
BEACH
400-500
> 1500
Ait AH HANCOCK fOONOAIION
US.C
HAMQH tNVltONtt!Nl*| PKOlECTS
Figure 10. Distribution of total number of eqqs/m^ in the Los
Angeles-Long Beach Harbors in 1978.
-------
SCill IN MlllS
WILMINGTON
LONG
¦ EACH
SAN
PEDRO
CAlRlllO
BEACH
AUAM MANCOC H FOUNDATION
U 5 C
%naveas%ng
relative
abundance
HaMOU tHVUONMfNUi MOJiC 1%
Figure 11. Stations showing highest abundances of engraulid larvae,
H
H
ro
K)
H
VD
-------
tCAit IN «IHt$
WIIMINCION
LONG
BEACH
SAN
HDIO
tnareastng
relative
abundance
CAitlLlO
If ACM
HANCOCK (FOUNDATION
use
iNV
-------
SCAlt IN Mllli
WILMINGTON
IONG
• EACH
16,
*5
*4
*2
zncreasing
relative
abundance
CAMIUO
IfACH/
AUAN HANCOCK FOUNDATION
HAHtOB INVHQNMf N TAt HOjtCTI
Figure 13. Stations showing highest abundances of gobiid larvae.
-------
SCALl IN MlICi
LONG
IEACH
SAN
PIDttO
CABllllO
•fACM
AUAN HANCOCK rOUNDATION
US.C
marea&xng
relative
abundanae
A	 H»UQI tNVt»OWM(HIAl HOif Cf i
Figure 14. Stations showing highest abundances of Hypaoblennius
sp. larvae.
-------
IIP 25
223
Table 1. Monthly sampling regime in Los Angeles-Long Beach
Harbors in 1978 according to stations and to sam-
ling and quantification methods.

J
F
M
A
M
J
J | A
S
0
N

2
B*
B*
B
B/
/s°
B^So
B*
f s°

B/
' s*
B*
S*

B/
XS*

3



B/
/gO
B^s°
b;
' so

B/
'S*
B7
/S*

B/
/s*

4
B*
B*
B
B/
/s°
/s°
B/
SO

B/
S*
B/
S*

B/
s*

5



B/
/ s°
B/
/SO
B/
/ s°

B/
/s*
B/
/s*

B/
'S*

6
B*
B
B
B/
/s°
B}
s°
b;
/s°

B/
'S*
B/
' s*

B/
's*

7
B*
B
B
B/
/So
B/
' so
B/
7s0

B/
'S*
B/
'S*

B/
xs*

15
B*
B*
B
B/
/so
B*
/ so
B/
' so

B/
' s*
B/
/S*

B/
' s*

16
B*
B*
B
B/
s°
O
CO
CQ
B/
' so

B/
7S*
B/
/ S*

B/
's*

17
B*
B*
B*









21
B*
B*
B









day	night
B = bongo nets
S = surface tows
o = identified but not quantified
* = identified and quantified
-------
224
IIP 26
Table 2. Taxonomic classification of larvae and eggs collected
in Los Angeles-Long Beach Harbors in 1978.
Atherinidae
unidentified
Blenniidae
Eypsoblennius
Bothidae
Pavali ohthy s/Xy s treuvy a
Cithariahthys
Carangidae
Seriola/Traahurus
Clinidae
unidentified
Cottidae
Clinooottus type
Scovpaenichthys marmoratus
unidentified
Cynoglossidae
Symphurus atriaauda
Engraulididae
Engraulia mordax*
Gobiesocidae
Gobiesox vhessodon?
Gobiidae
unidentified
Labridae
Oxyjulis ealifovnica
Merlucciidae
Merluca-Lus productus
Myctophidae
Stenobraehius leuaopsarus
Triphoturus mexicanus
Ophidiidae
? Chilara taylovi
? Otophidium scrippsi
Pleuronectidae
Eypsopsetta guttuiata
Pleuronichthys ritteri
Pleuronichthys vertiaalis
Pomacentridae
Chromis punatipinnis
Eypsypops rubioundus
Sciaenidae
Cheilotrema saturnum
Genyonemus lineatus
Seriphus politus
Scorpaenidae
Sebastes I
Sebastes II
Serranidae
Paralabrax
Sphyraenidae
Sphyraena argentea
Stromateidae
Pepri lus 3i.milli.mus
Synodontidae
Synodus lucioceps
Unidentified
Yolk sac larvae
* possibly other Engraulias included
-------
IIP 27
225
Table 3. Abundance of eggs and larvae from Los Angeles-
Long Beach Harbors in 19 78.
Taxon	# Captured #/100m3/yr / * occurre§S
Genyonemua lineatus
5001
108
,413
15
4
Engraulididae
2567
41
,034
40
9
Gobiidae
665
9
,243
39
6
Hypsoblennius
502
2
,930
38
9
Seriphus politus
92

203
10
3
Symphurus atricauda
73

178
5
1
Sebastes I
54

888
11
3
Stenobraohius leuaopsarus
34

596
6
2
Paralabrax
33

76
4
2
Paraliah thys/Xystveurys
27

402
9
4
Pleuroniahthys rittevi
23

357
8
4
Oxyjut-is californioa
23

56
1
1
Cithariahthy s
19

176
9
5
Sphyraena argentea
18

44
3
2
Pleuronichthys vertiealis
18

309
10
4
Gobiesox rhessodon?
15

174
9
3
Clinidae
11

124
10
6
Unidentified
11

98
6
2
Yolk sac
7

14
5
2
Atherinidae
7

—
4
1
Cottidae
6

99
5
2
Chvomis punatipinnis
6

13
4
1
Cheilotrema saturnum
6

13
3
1
Peprilus simillimus
5

7
3
2
Hypsypops rubioundus
5

45
4
2
Hypsopsetta guttulata
5

12
4
1
Merlucaius produatus
4

59
2
1
?Chilara taylori.
3

20
2
1
Clinoaottus type
3

35
3
2
?Otophid-ium sarippsi
2

4
2
1
Soorpaeniahthys mavmovatus
1

10
1
1
Seviola/Tvaahuvus
1

2
1
1
Triphoturus mexiaanus
1

10
1
1
Sebastes II
1

16
1
1
Synodus luoioceps
1

—
1
1
Engraulid eggs
316


13
7
Other eggs
22,913


56
8
-------
Table 4. Ranked order of ten most abundant species of fish and larvae.
1978	1974
LARVAE	ADULTS	LARVAE	ADULTS
Genyonemus lineatue
Genyonemus lineatue
Engraulididae
Genyonemus lineatus
Engraulididae
Symphurus atriaauda
Hypsoblennius
Engraulis mordax
Gobiidae
Ci tharichthys
stigmaeus
Sciaenidae
Symphurus atriaauda
Hypsoblennius
Seviphus politus
Sebastes
Ci thariohthys
stigmaeus
Seviphus politus
Engraulis movdax
Gobiidae
Seriphus politus
Symphurus at pi cauda
Phanerodon furcatus
Clinidae
Cymatogaster aggregata
Sebastes I
Synodus lucioceps
Chromis
Phanerodon furcatus
Stenobrachius
Paralichthys
californicus
Cottidae
Porichthys myriaster
Pai'alabrax
Hyperprosopon
argenteitm
Paralabrax
Lepidogobiu3 lepidus
Pavalichthys/
Xystreuvys
Sebastes dallii
Pleuronichthys
vertioalis
Sebastes miniatus
-------
IIIA
227
MONTHLY STANDING STOCK MEASUREMENTS OF BACTERIOPLANKTON
AND PHYTOPLANKTON IN LOS ANGELES HARBOR AND
SOUTHERN CALIFORNIA COASTAL WATERS
INTRODUCTION
Current interests concern the relationship between the
organic material in sewage and cannery waste effluent discharges
into the outer Los Angeles Harbor and the bioenhancement of
those waters. Microbes play a key role in the cycling of this
material into the food webs which characterize this bioenhance-
ment. A principal role of marine bacteria is the respiration
of organic compounds and the consequent regeneration of
inorganic nutrients. This activity makes the substrates of
primary production available to photosynthetic organisms in
the harbor. Marine microbes also initiate an important food
web by their•assimilation of dissolved organic matter (hetero-
trophic production). Their cells are then made directly avail-
able as food to higher trophic levels. Therefore, to under-
stand these ecologically important members of the marine environ-
ment better, a program to determine the monthly standing stock
measurements of bacterioplankton and phytoplankton in outer
Los Angeles Harbor was undertaken in September 1977. The outer
harbor then received fish cannery wastes and secondary effluent
from the Terminal Island Treatment Plant (TITP). The cannery
effluents were diverted to TITP in October-December 1977,
reducing organic nutrient input to the harbor substantially.
The effects of this reduction on the bacterioplankton popula-
tions was examined during the course of our study.
METHODS
The Acridine Orange Direct Count (AODC) method was used to
enumerate bacterioplankton. All water samples were prefiltered
through 203 jim mesh zooplankton net after collection in sterile
one-liter Niskin water samplers. An appropriate volume of
sample (one which yielded approximately 30 bacteria per field
when counting) was mixed with 0.5 ml borate buffered formalin
(100% formalin saturated with boric acid), 0.5 ml acridine
orange solution (50 mg per L stock), and 0.2y filtered sea
water (FSW) yielding a final 5 ml sample which was 5% formalin
and 5 mg per L acridine orange. After 3 minutes this mixture
was filtered through the appropriate porosity Nuclepore membrane
filter or directly onto a wet (with FSW) 0. 2\i 25 mm diameter
black Sartorius membrane filter (-10 cm Hg vacuum). The filter
was then rinsed with 5 ml FSW. A drop of immersion oil, the
filter, another drop of oil, and coverslip were placed on a
glass slide.' This was stored in the dark at 5°C. Ten fields
per filter were counted using epifluorescent illumination and
lOOOx magnification. A mean and standard deviation for the
number of bacteria per filter was calculated and this number
-------
228
IIIA 2
converted to bacteria per ml ± one standard deviation.
Autofluorescent particles were enumerated from August
through December 1978 using an identical slide preparation
technique, but without acridine orange staining of the water
sample.
Phytoplankton biomass estimates were courtesy of J. SooHoo
{Chlorophyll a measurement), T. Sharpe (ATP measurement), and
R. Ruse (floristics).
RESULTS AND DISCUSSION
The concentration of live bacteria (cells*L~l) was deter-
mined by the acridine orange direct count (AODC) method (Daley
and Hobbie, 1975), and a biomass estimate (ygC*L-1) (Ferguson
and Rublee, 1976) of the standing stock was made monthly for
1978 for water samples taken lm below the surface at four sta-
tions (A2, A7, A12, B9) in the outer Los Angeles Harbor (Figure
1) and one station (AO) in the coastal waters outside the harbor
breakwater (Figure 2). The range over the year for stations in
the harbor is 1.6 X 108 cells L~l (1.3 ygC'L"1) to 55 x 10®
cells'L-1 (42.8 ygC'L-1), while the annual range in standing
stock outside the breakwater is only 1.6 x 108 cells*L-1
(1.3 ygC'L !) to 18 x 10s cells*L 1 (14.0 ygC*L M. The
monthly bacterial standing stock in harbor waters averages
2.5 times that found in coastal waters. All stations show two
seasonal blooms of bacteria — one in late spring and another
in early fall.
Phytoplankton biomass was estimated monthly by three inde-
pendent methods: 1) chlorophyll a measurement (J. SooHoo,
personal communication; Figure 3); 2) ATP content of particles
(T. Sharpe, personal communication; Figure 4); and 3) direct
count of phytoplankton and microzooplankton (R. Ruse, personal
communication; Figure 5). Collectively (Figure 6), these data
show elevated phytoplankton biomass levels for various stations
from April through September, with maxima occurring at different
stations in June (A7), July (A2, A12, B9), August (AO), and
September (A2, A12, B9). The average annual range is.100 to
1700 jigC L-1 for harbor stations and 100 to 1000 ygC L-1 for
station AO. The phytoplankton bloom in late spring coincides
with the bacterial bloom at that time. The late summer phyto-
plankton bloom is followed by an early fall bacterial bloom.
The bacterial blooms are correlated with times of increased
levels of dissolved and particulate organic materials resulting
from: 1) the high phytoplankton standing stock, 1) excretion
by phytoplankton, and 3) grazing and excretion by zooplankton.
The natural microbial population was size-fractionated
each month by passage of water samples through various porosity
Nuclepore membrane filters (Figure 7-15) . The harbor water
populations are composed of a smaller percentage of small cells
-------
IIIA 3
229
averaging 112% - 27 (1 S.D.) <5y, 90% ± 21 (1 S.D.) 
-------
230
IIIA 4
Before the discharge was diverted to the Terminal Island Treat-
ment Plant for secondary treatment (TITP), bacterial concentra-
tion at the various harbor stations was directly related to
proximity to the site of disposal near station A 7 (Figure 1).
Station A 7 contained 1648 X 108 cells'L-1 while the other three
harbor stations (A2, A12, B9) contained 237-308 x 10a cells'L-1 .
Discontinuation of cannery discharge resulted not only in a
reduction in total bacterial numbers at all stations, but also
in an equalization in the number of bacteria among the four
harbor stations. A 27-fold difference in bacterial concentra-
tion inside (A7) versus outside (AO) the breakwater in Septem-
ber, 1977 was reduced to a 3-fold difference in September, 1978.
It is hypothesized that differences observed between 1977 and
1978 were due to the discontinuation of cannery effluent
disposal near station A7. The TITP effluent had already been
converted to secondary treatment prior to this study.
Cannery effluent is rich in dissolved organic nutrients,
and could support the large population of microheterotrophs
found near station A7 in 1977. When discharge was discontinued,
nutrients were no longer available to support a large bacterial
population. Therefore, when all effluents were converted to
secondary treatment in 1978, 1) the total number of bacteria
at all stations decreased significantly, 2) the bacterial
concentration at A7 was reduced to levels comparable with the
other harbor stations, and 3) the large bacterial concentra-
tion difference between the harbor stations and station AO was
reduced significantly. Even though data are available for one
season only, the magnitude of the change in standing stock of
bacterioplankton observed is not believed to be due solely to
natural year-to-year fluctuations.
A 10-fold increase in marine bacteria which occurred at
all harbor stations from June through October, 1978, originally
ascribed to a seasonal pattern related to phytoplankton biomass,
warrants reconsideration in light of information recently made
available concerning TITP effluent composition and flow-rate.
A TITP malfunction from June through August, 1978 resulted in a
10-fold increase in the levels of suspended solids and BOD
values in the effluent. This effluent, then, was potentially
of sufficient quality and quantity to generate the observed
microbial bloom. Thus such changes in the microbial population
apparently are excellent indicators of changes occurring in
effluent composition.
Note, however, that a parallel change in bacterial numbers
was observed at station AO, which is outside the harbor and
might be considered as "upstream" of the receiving waters leav-
inq the harbor. Therefore, a seasonal bloom of bacteria and
the TITP malfunction may have occurred simultaneously, jointly
affecting microbial biomass in the harbor, or tidal flushing
may distribute the nutrients outside the harbor when levels
are high enough in the effluent. A second year of monitoring
microbial populations in the harbor may allow one to distin-
guish between these alternatives.
LITERATURE CITED See Section VI
-------
Wll MINGTON
LONG
BEACH
Ml
V/
SAN
PEDRO
LONG
BEACH
^HARBOR
SAN PEDRO BAY
PT FERMIN
Scale in Miltj
2
Figure 1. Monthly Monitoring Stations, 1978- 1979. The lined area is approximately 10 km .
M
H
H
>
Ui
N)
U>
-------
232
IIIA 6
AO
30-
-38
20-
-26
10-
-13
A2
30-
-38
20-
-26
10-
-13
A7
30"
V
V
1'°-
K>~
-38 m
-26
rl 3
A12
38
30-
20-
10-
B9
30-
-38
20-
-26
10-
-13
Winter
Spring
Fall
Summer
1978
Figure 2. Bacterial Concentration (10* Cells/1.) and
Biomass (/tg C/L) from Total AODC
-------
IIIA 7
233
AO
2-
A2
2-
1-
A7
-<
s
V
o>
E
A12
1-
B9
2-
1-
1978
Figure 3. Biomass (mg C/L) from Chlorophyll g x 75
Measurements
-------
234
IIIA 8
2
1
2
1-
at
E
2
1-
2
1
AO


A2


A7
A12
•-	•	»	

B9

—	•
i i
J F M
i I 1 !
A M J J
	 I 1 ] I 	
A S O N C
1978
Figure 4. Biomass (mg C/L) from ATP Analysis >1/4
-------
IIIA 9	235
1978
Figure 5.
Biomass (mg C/L) from Phytoplankton
AND MICRQZOQPLANKTON COUNTING
-------
236
IIIA 10
AO
2-
A2
A 7
i >—
A12
B9
1-
1978
Figure 6. Biomass (mg C/l) Composite of Three
Phytoplankton Methods
-------
IIIA 11	237
60
0	0 An Ouliidt Harbor Station
Outtr I. A. Harbor Stations
50
Z 40
O 30
20
0
T
T
T
T
T
T
T
T
T
T
T
T
T
T	I	I	I	I	!	!	1 T I " I T r » "" \	III!
S O N D i FMAMJJASONDJ FM
i	i
X97*	1979
Figure 7.
Total oo.2>i) Bacterial Concentration
-------
238
IIIA 11
O	q A0 Outsid* Harbor Station
Ct	A A2 "1
* * fOuter L.A. Harbor Stations
50-
40-
30-
lO-
~r
M
t	1	r
A M J
J A S O N D
1978	1979
Figure 8.
Acridine Orange Direct Count tAOOC) (<5ji)
Bacterial Concentration
-------
IIIA 13
239
• (230>
A (93)
60-
O	0 Ag Outside Harbor Station
Outer L.A. Harbor Stations
~	DA
30-
O 40 -
30 -
30 -
1979
1978
Figure 9. Acridine Orange direct Count cAODC)
(dp) Bacterial Concentration
-------
240	IIIA 14
Outsido Harbor Station
Station*
Ogttr LA. Hai
SO-i
40-
E
s
•	30-
k
•
*•
V
1
o
«»
20-
*
*o-
Figure io.
acridine Orange Direct Count (AODC)
Bacterial Concentration
( <0 . 6fl)
-------
IIIA 15	241
• (US)
+ total Bacteria
60-
50-
E
v
«>
W
O
A
20-
Figure u.
Station AO Acridine Orange Direct Count (AQDC)
-------
242	IIIA 16
s
60 n
50-
40-
30-
O
X JO-1
10-
+	Total Bactaria
•	<5 M
¦	<1.0 M
A	<00/4
—r~
O
N
1978
1979
Figure 12.
Station A2 Acridine Orange Direct Count (AODC)
-------
IIIA 17	243
t Total Bacteria
• <5 Jl
50-
30-
20-
K>-
T
T
197«	1979
Figure 13.
Station A7 Acridine Orange Direct Count 
-------
244	IIIA 18
Total Bacteria
<*A
* 1.0 /i
< 0.6/c
50-
40-
30-
ao-
K>-
T
T
T
197$	1979
Figure 14.
Station A12 Acridine Orange Direct Count (AODC)
-------
IIIA 19	245
+(237)
+ Total Bacteria
* <06
50-
30-
lO-
I97«	1979
Figure is. Station B9 Acridine Orange Direct Count 
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246
IIIA 20
AO
400-
200-
A2
400-
200-
A7
A12
400-
200-
B9
400-
200-
1978
FIGURE 16. Biomass (/ug C/L) from ATP Analysis
-------
IIIA 21	247
AO
30-
20-
10-
A2
30-
20-
10-
A7
30-
\
U
m
20-
10-
A12
30-
20-
lO-
B9
30-
20-
10-
1978
Figure 17. Biomass 
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248
IIIA 22
% of mai.
D€PTH CSixo Fractions)
Figure 18. a/17/78 Acridine Orange Direct Count
(AODC)iA2 Bacterial Size Fractions vs. Depth
-------
IIIA 23
249
AO
2-
A2
2-
A7
2-
A12
100-,
80-
B9
20-
Filfer Porosity(^im)
Figure 20.
1978
Figure 19. Biomass (/ig C/L) from Autofluorescence
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Intentionally Blank Page
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IIIB
251
THE INGESTION AND UTILIZATION OF LABELED MARINE BACTERIA
BY HIGHER TROPHIC ORGANISMS FROM
LOS ANGELES HARBOR AND CALIFORNIA COASTAL WATERS
INTRODUCTION
A considerable research effort has been devoted to determine
the role of raicroautotrophs (diatoms, dinoflagellates, monads,
etc.} as a nutritional resource for higher trophic levels (zoo-
plankton, suspension and deposit feeders) in aquatic ecosystems.
It is now well known that these organisms play a significant
role and are considered to be the primary food base for many
ecosystems. However, Fenchel and J^rgensen (1977) recently
estimated that 40% to almost 100% of the carbon fixed in primary
production is utilized by the secondary producers or micro-
heterotrophs (bacteria, yeasts, fungi and Protozoa),
depending on the ecosystem in question. Indeed, it does seem
reasonable that organisms which rapidly cycle dissolved carbon
and produce particulate biomass, such as bacteria, will not go
unexploited as a food resource for higher trophic organisms.
The role of the detrital food web may be more significant in
ecosystems which are either organically enriched or deficient
in a necessary component for photosynthesis, i.e., light, nitrogen
or phosphate. Pomeroy (1974) has recently pointed out that in
the open ocean microheterotrophs play a highly significant role,
both in the nutrition of higher trophic organisms and in their
long known role in nutrient regeneration.
It is now becoming well established that bacteria serve as
a nutritional resource for many aquatic organisms, including
planktonic and benthic feeders. The early workers Doflein and
Reichenow (1928) stated that some Protozoa, and ciliates in
particular, feed upon bacteria; it seemed likely to them that
free and attached bacteria were consumed and metabolized in
planktonic ecosystems. More recently several investigators
(Zobell and Felthan, 1937; Fenchel, 1969, 1972, 1975; Barsdate
et al., 1974; and J^rgensen, 1966) have obtained good evidence
that bacteria do play a substantial role in the nutrition of
deposit and filter feeders. Wavre and Brinkhurst (1971) have
demonstrated experimentally that bacteria are digested from the
bolus as it passes through the gut of tubificid oligochaetes.
Duncan et a_l. (1974), by means of a simple radioassay, showed
that bacteria are ingested and assimilated by the aquatic nema-
tode Pleatus palustris. Sorokin (1973, 1978) has reported that
filter feeders in coral communities, such as sponges, ascidians,
sabellid polychaetes, and oysters, are capable of filtering
bacterioplankton from the water. He also found that some species
of coral, gastropods, and holothurians were capable of ingesting
and assimilating bacterial biomass.
-------
252
IIIB 2
The following report is the result of investigations conducted
to develop a standard technique which can be used to obtain
detailed quantitative information on the flux of bacterial carbon
through bactivorous organisms. The method was developed and
tested using a marine bacterium as food source and a bactivorous
ciliate, Euplotes sp., as a predator which utilizes bacteria as
a source of carbon and energy. Both organisms were isolated from
the Los Angeles Harbor. This technique was then used to
determine whether bacteria are ingested and metabolized by:
Euplotes sp. (Protozoa), Neanthes avenaaeodentata (Polychaeta),
Maaoma nasuta (Bivalvia), Mytilus edulis (Bivalvia), and natural
assemblages of microzooplankton (5-203 }im) from five Los Angeles
Harbor stations.
MATERIALS AND METHODS
Culture Preparation
The clone of rod-shaped bacteria used for labeling was grown
and isolated on Lib-X agar medium. The 14C-labeled bacteria were
prepared by growth in an organic-free seawater medium composed of
Rila Utility Marine Mix and 50 mg of sodium nitrate in one liter
of distilled water, pH 7.8, to which was added fifty microcuries
of uniformly labeled 14C-glucose (NEC 042A-lmCi, 37.1 mg in 10 ml)
per 100 ml of sterile medium. The culture medium was inoculated
by transferring cells from agar medium (Lib-X) using a bacterio-
logical needle. The culture was grown on a shaker table at 18°C
for a minimum of five days; two days were required to reach sta-
tionary phase, after which the cells were maintained for at least
three days in starvation phase to reduce their metabolism of endo-
genous l^C storage pools. Before the experiments the labeled cells
were collected on a 0.2 jam pore size membrane filter (Nuclepore)
by gentle vacuum pressure (<7 mmHg), rinsed, and resuspended in
chilled seawater medium by vortexing.
Unlabeled bacteria were prepared by growing the same bacterial
isolate in a Lib-X medium containing per liter: Rila sea salts
(40 g), glucose (1.0 g), trypticase soy broth (2.3 g), and yeast
extract (1.2 g), pH = 7.8. The culture was grown at 18°C for 24 h,
centrifuged at 10,000 rpm for 10 min, rinsed and centrifuged twice.
The pellet was resuspended in unsupplemented artificial seawater
and incubated as before for at least 20 h. The cells were centri-
fuged and washed again before use. The labeled and unlabeled cells
were added to the experimental medium to a density which reflects
the range of natural standing stocks of bacterioplankton in Los
Angeles Harbor. The labeled cells usually comprised less than 10%
of the total bacterial population.
Euplotes sp. laboratory studies
Euplotes sp. was isolated from a Los Angeles Harbor water
sample by means of an enrichment culture technique. The ciliate
stock cultures were maintained in a seawater medium with the
-------
IIIB 3
253
addition of brown rice and the natural microflora at 18°C in
constant light.
Before the initiation of experiments the culture and medium
were passed through a 100 ;im mesh to remove large detrital parti-
cles. The ciliates were then concentrated four times by a
gentle reverse filtration process. This process removes water
from inside a column after it passes through a 10 )im mesh which
covers the end of the column and is positioned at the bottom of
the culture vessel. The water passes through the filter into
the column to reach equilibrium with the outside of the column
and is removed by a peristaltic pump.
To demonstrate the relationship of grazer concentration to
the rate of removal of bacteria from the medium, four dilutions
of the ciliajpe suspension were prepared (20.5, 51, 102, 205
ciliates'ml- ) using the ciliate medium previously removed. Since
the natural microplankton was already present, only the labeled
bacteria were added. Ingestion, respiration and excretion assays
(methodology described below) were conducted at 0, 40, and 90
min after initiation of the experiment.
In an experiment to illustrate the relationship between
bacterial concentration and grazing rates, the ciliate concen-
tration was held constant and the bacterial concentration was
varied (1.28, 1.86, 4.02, 7.85 X 106 bacteria-ml"1). Unlabeled
bacteria inoculated with the 1.0 Jim filtrate from a Euplotes sp.
culture were prepared in the rice-seawater medium and stirred
vigorously for 96 h at 18°C. These bacteria were passed through
a 25 p mesh and 0.5, 10"^-, 0.5 X 10 , 10~2, and 0.5 X 10~2 dilu-
tions were made with 0.2pm filtered medium. The labeled bacteria
were added to each of the dilutions in a constant amount and the
experiment was initiated when inoculated with a constant volume
of ciliate suspension. The culture was sampled at 0, 40, and 90
min. Ciliate concentration was determined microscopically from
four 1 ml replicates, samples were fixed with Lugol's iodide and
were counted under a dissecting microscope. Since the labeled
bacteria were added in the same amounts, the bacterial concentrate
of each dilution had to be determined directly by means of a
Petroff-Hauser cell counter and phase contrast microscope.
Benthic invertebrates studies
Experimental specimens of uniform size were selected and
starved in beakers of filtered seawater for a minimum of 4 8 h
before the experiment. The medium for Neanthes arenaseodentata
was decanted twice daily to remove fecal pellets. The valves of
the Mytilus edulis specimens were brushed, scraped and dried before
the experiment to remove any epizoa that might interfere with the
results. The individual experiments with invertebrates were
modified from the basic plan to accomodate the particular
biological requirements of the test organisms. These conditions
are described below.
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254
IIIB 4
20 October 78 - Neanthes arenaeeodentata
The fine sand sediment used for this experiment was oxidized
with 30% hydrogen peroxide and rinsed thoroughly to remove most
organic material. This procedure was carried out with the idea
of reducing bacterial growth and metabolism but was later found
to have no appreciable effect, so it was discontinued in sub-
sequent experiments. Rila Utility Marine Mix with a suspension
of labeled and unlabeled bacteria was used as the assay medium.
Each specimen was placed in a 100 ml serum bottle after 5 ml of
sediment and 20 ml of medium were added; incubation was in
darkness at 17°C. Replicate samples lacking test specimens were
prepared for bacterial background controls. Excretion, respira-
tion and ingestion assays were made at 0, 24 and 26 h.
20 October 78 - Maooma naauta
The methods and materials were the same as described above,
except the specimens were placed in 125 ml erlenmeyer flasks and
40 ml of medium and 10 ml of sand were used.
27	October 78 - Neanthes arenaeeodentata - Pulse chase experiment
The pulse chase experiment was designed to observe the meta-
bolic fate of ingested, labeled bacteria during long incubation
times. The worms were given a pulse of labeled bacteria for 48 h,
rinsed and transferred to medium with unlabeled bacteria (chase)
and allowed to feed for 48 h. It was postulated that the absence
of observable worm respiration may be due to slow gut passage
time and high bacterial background respiration, so the experiment
was designed to reduce bacterial background respiration and allow
sufficient gut passage time to observe the eventual respiration
of ingested bacterial carbon.
In this experiment no sediment was used. Unlabeled bacteria
were added to 800 ml of artificial seawater and the labeled cells
were added to 500 ml of this medium. Duplicate sets of serum
bottles were prepared, one set for initial pulse uptake, and the
other set for the subsequent experiment containing 20 ml of labeled
medium and one specimen. Ingestion, excretion, and respiration
assays were taken at 0 and 48 h. Worms were removed from the
duplicate set of bottles and rinsed in filtered sea water and
then placed in new bottles with 20 ml of unlabeled bacterial
medium. All parameters were then assayed after another 48 h of
incubation in darkness at 17°C.
28	November 78 and 8 December 78 - Mutilus edulis
After 72 h of starvation the specimens were placed in 125 ml
erlenmeyer flasks with the labeled bacterial medium which was
prepared from filtered sea water. The first experiment employed
80 ml of medium and all parameters were assayed at 0, 15 and 41 h
after incubation at 15°C darkness. The second experiment
-------
11 IB 5
255
employed 60 ml of medium and all parameters were assayed at 0,
2, 5, and 24 h. Valve length of Mytilus was determined by
measuring the left valve after dissection.
Studies on natural bactivorous plankton populations - 2 August 78
An experiment was designed to observe the possible relation-
ship between bactivorous rates of natural microzooplankton
(5-203 pm) and natural bacterioplankton standing stocks. Samples
were collected on 2 August 1978 from stations AO, A2, A7, A12 and
B9 (Figure 6). The samples were passed through 203 jam mesh and
then 25% of the liquid sample volume was removed by reverse fil-
tration (discussed above) and passed through a 0.2 jam membrane
filter (Nuclepore). The labeled bacteria were added to this fil-
trate and the preparation was returned to the original sample.
It was thus possible to avoid tne problem of substantially chang-
ing the in situ bacterial concentration while adding the labeled
cells. The kinetics of ingestion, respiration and excretion were
determined by assaying at 0 and 90 min for stations AO, A7, A12,
and B9 and at 0, 15, 30, 45, 60, 120, 180, 240, and 360 min at
station A2.
Each station had a characteristic bacterial standing stock
which was determined monthly by means of epifluorescent counting
by D. Krempin? those values are used in this study.
Measured parameter definitions and procedures are detailed
in the following paragraphs.
Respiration. Respiration of bacterial carbon was determined
by the amount of -^CC^ collected after correction for bacterial
background and T^ blank. It is assayed by the methods described
by Hobbie and Crawford (1969). Small organisms are placed in
100 ml serum bottles sealed with rubber stoppers and the larger
organisms in 125 ml erlenmeyer flasks sealed with rubber
stoppers and parafilm. Each data point represents the rate of
a single specimen or sample. This sample cannot be used to
determine any other parameter, because of the acidification step
required by the method. The calculations of respiration rates
employed are as follows:
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256
IIIB 6
Rb = (Rbx - V
Rg = 2 collected at time point 0 - DPM
14
= bactivore's respiration rate of CC>2 - DPM/hour
14
RgX= total CC>2 collected at time point X - DPM
X = time point - hours
Excretion
Excretion is considered to be the flux of dissolved organic
carbon-14 into the medium due to biological activity. Bactivore
excretion is determined by subtraction of bacterial excretion
of 14C. It is determined by removing 5 ml of medium from
designated vessels, passing it through a 0.2 ;am Nuclepore filter
and collecting the filtrate. The filtrate is then acidified to
pH 2.0 by addition of HC1 and agitated for 20 min to evolve
the dissolved COj. A 1 ml aliquot of the filtrate is placed
in 10 ml of Aquasol scintillation fluor and the radioactivity
is measured. The equations employed are the following:
Eb - Ebx - Eo
Eg " V(Egx " Eo " V'*
14
= DOC excreted by bacterial controls - DPM/ml
E^x= bacterial control DOC-'-4 collected at time point X - DPM/ml
E^ = DOC^ collected at time point 0 - DPM/ml
14
E = bactivore's excretion rate of DOC - DPM/hour
E = total DOC14 collected at time point X - DPM/ml
gx
V = experimental medium volume - ml
X = time point - hours
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IIIB 7
257
Specific Activity
Specific activity is defined in this study as the total
number of bacteria represented by one disintegration per minute
(DPM) of carbon-14.
Bacterial numbers in the medium are determined by fixing
an aliquot with Lugol's iodide and counting cell numbers in a
Petroff-Hauser cell counter on a Zeiss phase contrast microscope.
The 14C associated with the labeled bacteria is determined by
collecting a 5 ml aliquot on a 0.2 jim filter at 0 h/ rinsing
twice with chilled sea water, drying and counting on a Beckman
LS-100 Scintillation Counter. Specific activity is calculated
as follows:
SPAC = C/A/v
a
A = activity of 0.2 jam filter retentate - DPM
C = bacterial concentration - cells/ml
SPAC = specific activity - bacteria/DPM
v = volume of medium filtered for A - ml
a
Ingestion Rate
Ingestion rate is the total number of bacteria or amount
of 14C taken in by the bactivore divided by the time period of
incubation after being corrected for metabolic fluxes and T ,
blank. It represents the rate at which the bactivore removes
bacteria from the medium.
The activity of the microzooplankton (including Euplotes sp.
studies) was determined by radioassay of the 5.0 pm filter
retentate. At each time point 10 ml aliquots of the assay
medium were passed through 5.0 pm Nuclepore filters and rinsed
twice with 10 ml of chilled sea water, dried, and counted in
Toluene, PPO, POPOP scintillation fluor. The 0 h filters rep-
resent the bacterial background activity which is corrected for
metabolic fluxes and subtracted from the other time points.
The radioactivity associated with metazoans was determined
as follows: The specimens were removed from the medium and
placed in filtered seawater for a 10 min rinse to remove
extraneous labeled bacteria from metazoan surfaces. Soft-bodied
specimens were placed in scintillation vials with 1 ml of
Protosol (New England Nuclear) and homogenized.
The bivalves were opened and all tissue was removed with
a scalpel and placed in a scintillation vial with 1 ml of
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258
IIIB 8
Protosol and homogenized. Samples were allowed to digest over-
night and then were heated to 55°C for 30 min for final digestion,
To reduce color quenching 0.1 ml of 30% hydrogen peroxide was
added and the samples were heated for 30 min at 55°C. The
following day Aquasol scintillation fluor was added and the
samples were counted. Bacterial ingestion rate is calculated
as follows:
L = (Eb + Rb/Vr)/A/Va
Td - Fx - F,i <1 - L) + Eg + Rg
Ib - Id (SPAO/X
F^ = activity of bactivore at time point 0 - DPM
F = activity of bactivore at time point X - DPM
1^ = ingestion rate - bacteria/h
I. = total activity ingested, including metabolic cor-
rections - DPM
L = proportion of bacterial metabolic losses from blank
at time point X - O.XXX
V = volume of respiration assay - ml
RESULTS
Euplotes sp. laboratory studies
In the experiment to demonstrate the relationship between
grazer density and rates of bactivory, the bacterial concentra-
tion was 1.3 X 10 cells'ml"*•*¦ and 48% of these cells were
carbon-14 labeled. The ingestion and metabolic rates were almost
linear for the 90 min incubation period with a slight break
after 40 min. The data presented in Figure 1 and Table 1 are
based on the 40 min data points, because they are thought to be
more representative of the actual rates.
Figure 1 illustrates the linear relationship between grazer
population density and grazing rate. It can be seen that the
number of bacteria removed from the medium is directly propor-
tional to the number of grazers (Euplotes sp.) present in the
medium. The linear regression correlation to the data points
is 0.996 and is viewed as being significant. This observation
gives an indication of the reliability of the methodology in
determining bacterial grazing rates. In this experiment
bacterial concentration was probably not limiting during the
incubation period.
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IIIB 9
259
Table 1 illustrates that, in samples with a higher grazer
density, fewer bacteria per individual grazer are consumed.
This is probably due to fewer available bacteria in samples
which have stronger competition for resources and indicates
the importance of the relative grazer/bacteria concentration.
The data also reveal an inverse relationship between grazer
density and bacterial population turnover time. In other words,
the more grazers present the more rapidly bacteria will be
removed from the medium. Respiration and excretion data do
not reveal any significant trends related to grazer density.
Overall, approximately 12-16% of the ingested bacterial biomass
is used in predator energy metabolism, as indicated by the
respiration of bacterial carbon.
In a related experiment the grazer population density was
held constant and bacterial concentration was varied. Figure 2
illustrates the relationship between specific ingestion rates
and bacterial population density. The relationship seems to be
a hyperbolic function and analagous to that described by
Michaelis-Menten saturation enzyme kinetics. The last pair of
points seems to approach the saturating concentration, although
two sets of higher points are necessary to confirm this idea.
In the bacterial concentration range examined, the popula-
tion turnover time data displayed no significant trend related
to bacterial concentration according to data presented itt
Table 1. As presented in Figure 2, excretion and respiration
rates displayed a similar pattern of saturation kinetics, while
ingestion rates and excretion rates seem to be reduced at or
near saturation for ingestion. It is interesting to note that
in the most concentrated sample a ciliate is six times more likely
to encounter a bacterium than in the least concentrated sample
and the specific ingestion rate demonstrated experimentally was
5.5 times greater.
Benthic invertebrates studies
20 October 78
In experiments to demonstrate the uptake and utilization
of bacterial biomass, Neanthea arenaceodentata displayed no
adverse reactions to the chemically oxidized sand such as had been
observed earlier in thermally combusted sediments. The worms
formed burrow tubes and assembled mucus nets above the sediment.
C
Bacterial numbers were determined to be 4.6 X 10 cells/ml
which may be slightly low for the natural sediment-water inter-
face. The respiration assay failed to yield respiration above
bacterial background (see Table 2.). The excretion data reveal
that 87.8 to 99.6% of the ingested labeled bacterial biomass
was excreted. During the experiment these worms ingested 1.9
X 107 bacteria/day or approximately 20.6% of the available
bacteria. (text continued on p.12)
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TEXT TABLE 1
SUMMARY OF BACTERIAL INGESTION AND UTILIZATION BY Euplotes sp.
Sarrple
Incubation
Time
(min)
Ib/Ind
Ingestion Rate
(Bact/Indv/Min)
Mean	Range
Tt
Turnover
Time
C/Ib
(h)
Respiration Rate
of
Bacterial Biomass
% Ingested/Hour
Mean
Excretion Rate
of
Bacterial Biomass
% Ingested/Hour
Mean
Grazer Density
20.5 ciliates-nil
40
43.3
41.9-44.7
24.4


51
40
36.6
23.4-49.8
11.6
4.1
11.6
102
40
35.8
32.9-38.7
5.9
3.3
11.6
205
40
32.0
31.7-32.3
3.3
3.6
9.1
Bacterial Density






1.28x10^ oells'inl-^
70
20.0

130.0
3.9
17.7
1.86
40
33.5
28-39
114.0
7.3
42.0
4.02
40
84.0
75-93
75.2
5.2
46.5
7.85
40
110.0
105-115
112.2
14.8
40.9
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TEXT TABLE 2
SUMMARY OF BACTERIAL INGESTION AND UTILIZATION BY METAZOANS
No. of Size
Species	Date Specimens Range
Ingestion Rate
(Bact/Indv/Day)
Mean	Range
Ingestion-
Efficiency
(100 x Ingested/
Available)
%
Respiration Rate
of
Bacterial Biomass
% Ingested/Day
Mean	Range
Excretion Rate
of
Bacterial Biomass
% Ingested/Day
Mean	Range
Neanthes 10-20-78
avenaaeodenta ta
12 100-120mg 1.9x10' 1.7-2.1x10'
20.6
N.D.*
N.D.
93.7
87.8-99.6
(DNeanthes 10-27-78
arenaeeodentata
12
100'
-120mg 2.2xl09 1.5-2.8xl09 97
N.D.
N.D.
22.4
16.3-28.6
Maeoma	10-20-78 12 15~20nm 5.4xl06 4.3-7.8xl06 5.9	N.D.	N.D. 58.0 39.6-77.8
naauta
My til us
edulis
11-28-78
10
18-23nm
2.0xl08
1.1-3.2xl08
52.1

8.0
7.0-9.0
1.0
0.8—1.2
Mytilus
edulis
12-8-78
16
17-24ram
1.4x108
0.5-2.5xl08
16.5
•
5.0
2.8-8.6
N.D. **
N.D.
*N.D. = Not Demonstrated. Data failed to yield significant results above bacterial background.
Data for this experiment was not corrected for excretory or respiratory fluxes.
**N.D. = Not Demonstrated. Sanples lost due to defective scintillation fluor.
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262
IIIB 12
In a similar experiment which used Maeoma nasuta as the
test organism, the mud clams did not burrow into the sediment,
but remained lying on the surface; experimental conditions
apparently were not optimal. The specimens did, however, appear
to be pumping with their siphons. Respiration of bacterial
biomass was not demonstrated above background (Table 2) and the
specimens were found to excrete 39.6 to 77.8% of the ingested g
bacterial biomass. Also note that they ingested only 5.4 X 10
bacteria/day or approximately 6% of the available bacteria.
27 October 78
In the pulse chase experiment which utilized higher bacter-
ial numbers and an absence of sediment, Neanthes arenaoeodentata
was observed to form mucus tubes and mucus nets above them.
Bacterial concentration was determined to be 7.75 X 10^ cells/ml.
These specimens appeared to be very efficient at removing
bacteria from the water under the experimental conditions. They
ingested 1.5 to 2.8 X 10® bacteria/day (Table 2) which repre-
sents ingestion of approximately 97% of the available bacteria.
These data were not excretion-corrected because of technical
problems. Respiration of ^-^C02 above bacterial background was
not demonstrated for Neanthes arenaoeodentata.
Table 3 illustrates that the activity of the animal homo-
genate is substantially reduced after 48 h and excretion accounts
for approximately 13% of this reduction. Fifty-one percent of
the loss from the animal homogenate was not accounted for and
no significant respiration of ^-^002 was exhibited.
Text Table 3
27 Oct 78 Neanthes arenaoeodentata Bacterial Uptake Pulse-Chase
Experiment
Activity of Animal Activity of Flux
Homogenate	into DOC-^
Time	DPM	DPM
Activity
Unaccounted
DPM
Unaccounted
% Loss
0
48 h
126154
45048
16800
64306
51
28 November 78
The data for an experiment designed to test whether the
mussel, Mytilus edulis3 ingests and utilizes bacterial biomass
is presented in Figure 3 and Table 2.
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IIIB 13
263
The bacterial concentration was 4.8 x 10® cells/ml at the
outset of the experiment. This is within the range of the
standing stocks of bacterioplankton observed in the Los Angeles
Harbor. The data demonstrate that small Mytilus edulis
individuals effectively filter free bacterioplankton from the
water at a rate of 1.1 - 3.2 X 10^ cells/day (Table 2) which
represents an ingestion efficiency of approximately 52%. The
data also clearly illustrate that the ingested bacterial bio-
mass is metabolized (Table 2). During the experimental period,
approximately 8.0% of the ingested biomass was respired and
approximately 1.0% was excreted. Figure 3 shows the kinetics
of ingestion over the experimental period. It is readily appar-
ent that uptake is faster in the first 15 h and slows down in
approaching hour 40. The ingestion estimates appearing in
Table 2 are based on the average of all data and should be con-
sidered conservative; the actual ingestion rate may be higher.
The method employed yields an underestimate of feeding rates,
due to bacterial division and hence a dilution of the label.
8 December 78
The preceding experiment was repeated to test the effects
of bacterially-enriched conditions on uptake and utilization
rates. The bacterial concentration in this experiment was
higher (1.41 X 107 cells/ml) than in the previous experiment and
represented conditions in an organically enriched environment.
Table 2 presents data that show that the test organisms
(Mytilus edulis) ingested bacteria at a rate comparable to that
of the previous experiment (0.5 - 2.5 X 10® cells/day) and
removed approximately 16% of the available bacteria from the
suspension. Again, these figures are based on the average rate
from all data points. Ingestion kinetics ( Figure 3) appear
to be linear for the first 5 h and to decelerate for the next
19 h. Five percent of the ingested bacterial biomass was
respired by the test organisms (see Table 2). Excretion data
are not available, because the samples were lost due to defective
scintillation fluor.
Natural bactivorous plankton studies
These studies are designed to determine the seasonal varia-
tion in the rates at which bacterial populations are being
turned over by bactivorous plankton grazing in "Los Angeles
Harbor. The data presented in Figures 4 and 5 express grazing
rates in terms of bacteria ingested per unit volume and time,
because the species composition of the bactivores is not known.
Figure 4 compares the bactivore's grazing rate with the
bacterial standing stock at the five stations sampled. A direct
and significant (r = 0.945) correlation can be seen between the
two parameters. It should be noted that these rates may reflect
the grazing rates of individual bactivores and/or the population
density of bactivores. Table 4 presents ingestion and utiliza-
tion data for this study. The turnover times for the
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264
IIIB 14
bacterioplankton populations by bactivorous grazing ranges from
11.6 to 18.8 h. The rates of all five stations are comparable
and are not significantly different. The respiration and
excretion data show no trend related to bacterial standing
stock in the terms expressed in Table 4. This may be a
function of variability in bactivore population size, composi-
tion and activity. Bactivores with low grazing efficiency or
with strong competition pressures will have to expend more
metabolic energy than more efficient bactivores or those that
are in less competitive situations.
Text Table 4
Summary of Bacterial Ingestion and Utilization
by Natural Bactivorous Plankton Populations
Stn.
Bacterial
Ingestion Rate
Turnover
Respiration Rate
Excretion Rate of

Density
lb

Time
of
of

C
(bacteria x
C/Ib
Bacterial Biomass
Bacterial Bionass

(bacteria x
10b-IT

(h)
(% ingested / h)
(% ingested / h)

10y-L"l)
Mean
Range

Mean
Mean
AO
0.64
0.9
0.8-1.0
11.6
18.0
39.0
A2
1.79
1.6
1.4-1.7
18.8
40.0
44.2
A7
2.89
3.2
2.8-3.7
14.9
35.0
33.7
A12
1.17
1.4
1.3-1.6
13.7
10.5
14.5
B9
2.17
2.4
2.3-2.5
15.1
40.8
6.4
Figure 5 represents kinetics of bacterial ingestion by the
bactivorous plankton from station A2 during a 6 h incubation
period and the data points represent total ingested activity of
bacteria corrected for respiratory and excretory fluxes.
Although the correlation of the regression line (r = 0.87) is
not considered highly significant, it appears that ingestion
occurs at a fairly linear rate during the incubation period.
DISCUSSION
Euplotes sp. flaboratory studies
The experiments performed on laboratory cultures of
Euplotes sp., may illustrate some very basic and obvious ecologi-
cal principles ; i.e., there is a direct relationship between
the rate of removal of a food source from the environment and
the density of grazers, or that each grazer gets less of a
resource as competition for that resource increases. However,
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I.II.B 15
265
these ideas have been well borne out in other works and were not
the objective of this study. What this aspect of the study has
provided is a reliable method for quantification of the ingestion
and metabolic utilization of bacterial carbon by bactivorous
plankton, as is postulated to occur in Los Angeles Harbor or
other marine environments.
The study has illustrated that a monoculture of a bactivorous
plankter will remove bacteria from its medium at a rate directly
proportional to the concentration of bactivores. In effect,
the turnover time (T^) of a bacterial population appears to be
inversely related to bactivore population density. This in-
vestigation has also demonstrated that as competition for food
resources increases, caused by increasing bactivore population
density, each individual predator is able to capture less of
that resource. The data did not reveal any significant trends
in terms of metabolic energy expended related to competitive
pressure. This may be due to some inherent variability of the
methodology, or to the fact that this phenomenon may require greater
resolution or sensitivity than was designed into these experiments.
A relatively small portion (12-16%) of the ingested bac-
terial carbon was used for maintenance metabolism (respiration
and excretion). This may be attributed to the detritus and
organically enriched medium employed, which may also have been
utilized as a food resource and metabolized by the bactivores.
In addition, the carbon-14 incorporated into the bacteria is
probably mostly high molecular weight molecules, i.e., protein,
and is more likely to be used in anabolic rather than catabolic
processes by the predators. This would support Fenchel and
JeSrgensen's (1977) proposal that bacteria are an efficient and
energetically advantageous food source due to their low C:N ratio.
Ingestion kinetics which show resource saturation are also
suggested by this study. It follows that bactivores have a
maximal rate of grazing (Michaelis-Menten Vmax) and only attain
this rate when the food resource is present at some character-
istic concentration. At subsaturating bacterial concentrations,
the ingestion rate will vary with food availability. The
application of the Michaelis-Menten model to the data presented
here illustrated this phenomenon; i.e., initially the ingestion
rate increased rapidly with small changes in bacterial concen-
tration, and as resource saturation was approached the ingestion
rate declined. This suggested attainment of a constant rate
with no acceleration. The excretion and respiration data show
similar kinetics, and the rate of excretion appears actually to
decrease. It seems logical to assume that the metabolic
expenditure to capture food is inversely proportional to food
availability. The data presented in this study suggest that
respiration and excretion follow a similar pattern of satura-
tion kinetics, and that rates may even be reduced at or near the
saturation point due to optimization of capture success.
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266
IIIB 16
The results of these experiments have answered a few
necessary questions. First, they demonstrate that the method
employed can be used to demonstrate bacterial ingestion and
utilization by microzooplankton. They establish that bacteria
do play a nutritional role for Euplotes sp. The magnitude
or significance of this role has not been established and will
require further study. The techniques have also been useful
in demonstrating predator-prey or grazer-plankton relationships
in a closed system. This study also demonstrates the potential
of this assay for use with other organisms and in other circum-
stances .
The results of these studies indicated that a correlation
between bacterioplankton standing stock and rates of production,
and the feeding activity and standing stock of bactivorous or-
ganisms could be demonstrated in nature.
Benthic invertebrate studies
The results of the experiments with Neanthea arenaceodantata
demonstrate that these organisms do ingest bacteria collected
from both the sediment and the overlying waters. It appears
possible that not only do they ingest the sediment and digest
organic bacteria and matter, but they also may reingest the se-
creted mucus net after bacteria have been collected and colo-
nized on it. In the experiment which contained no sediment, it
appeared to be the only efficient means of gathering bacteria
in the absence of sediment. It is also possible that Neanthes
may ingest their fecal pellets after they have been colonized
by bacteria, as has been proposed by Frankenberg and Smith (1967).
However, this parameter was not considered in this study.
The absence of demonstrable respiration of ^CO is sur-
prising and hasn't been encountered in previous work. It may
be that the respiration rate of the worms is quite low and that
the labeled bacteria have enhanced respiration in the presence
of the worms, so that the total 14C02 collected is comparable
to the bacterial controls. Also, bacterial carbon, being large-
ly protein, may go into biosynthesis rather than energy metabo-
lism of the metazoans. Excretion values are significant and
do indicate that the bacteria are being metabolized.
,. The pulse chase experiment showed that 64% of the ingested
C was lost after 4 8 h. Excretion accounted for 13% of this
loss and the rest was not accounted for. It may be that this
C was contained in the fecal pellets. It cannot be determined
whether all the activity remaining in the animal homogenate is
actually assimilated bacterial biomass without knowing the gut
passage time.
The data from Maaoma nasuta suggest a relatively low
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IIIB 17
267
uptake rate per animal. The rather low ingestion rate could
be a reflection of two possibilities. The clam may not have
been feeding very actively, due to suboptimal sediment conditions
and/or their filtering apparatus may be inefficient at filtering
particles as small as free bacteria. The excretion data did,
however, demonstrate some uptake of bacterial biomass.
The experiments with young Mytilus edulis yielded the most
convincing results, supporting the hypothesis that bacterioplank-
ton play a nutritional role for some filter feeding benthic in-
vertebrates. Individuals 17-24 mm in length were capable of
ingesting 0.5 - 3.2 x 108 bacteria-day . The experiments also
yielded comparable metabolic activity results. This species
seems to be quite efficient at filtering free bacteria from sus-
pension. The different uptake efficiency figures among different
organisms (Table 2) are probably a function of the difference
in bacterial numbers in the two experiments. The difference in
the ingestion kinetics may be a result of possible oxygen lim-
itation in the second experiment due to higher bacterial numbers.
The method suffers from the disadvantage that all parameters
(uptake, ingestion, respiration, etc.) cannot be determined from
the same individual. In order to get a broader data base, ex-
periments should be performed using a greater number of uniformly
sized specimens as well as using different size classes of met-
azoans. Greater replication may yield more conclusive results.
The data presented herein lead to the conclusion that bacterio-
plankton can play a nutritional role for local benthic inverte-
brates in situ.
Natural bactivorous plankton studies
This study is the first in what will be a periodic sampling
program to correlate standing stocks and activity of bactivorous
plankton microheterotrophs in Los Angeles Harbor. This work is
designed to complement that which is being conducted on the de-
trital food web and the carbon cycle in Los Angeles Harbor.
The overall goal is to elucidate the dynamics of secondary pro-
duction in Los Angeles Harbor.
The data presented here illustrate a direct correlation
between bacterial grazing rates and bacterial standing stocks.
This indicates that standing stock and/or specific grazing rates
of natural populations of bactivorous plankton vary directly
with bacterial standing stock and production. It is postulated
that in an organically enriched marine system bacterial produc-
tion is enhanced, which in turn enhances bactivorous plankton
production, and so on. In this bacterial enhancement scheme,
which is the detrital food web, bacterial production is in equilib-
rium with bactivorous ingestion. It is thought that in situations
which are not nutrient- or oxygen-limited, this enhancement
scheme will be balanced. In defense of this statement, one can
look at the turnover time (Tt) of the bacterial population by
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268
IIIB 18
bactivorous grazing at each station compared to bacterial stand-
ing stock. The turnover time at station AO was 78% that of A7,
while the bacterial standing stock of station AO was 22% that of
A7. However, this may not be the case in organically enriched
waters, in which high rates of bacterial production, and hence of
high oxygen uptake, will inhibit the activity of other organisms.
No significant trend for metabolic activity or utilization
compared to bacterioplankton standing stock could be demonstrated
due to the heterogeneity of the bactivorous plankton populations.
It can be said that a substantial portion of the ingested bac-
terial biomass was used in energy metabolism (25-84%). Again,
this wide range can be attributed to heterogeneous bactivore
populations with varying feeding efficiencies and competitive
abilities. One expects fairly high metabolic rates for these
bactivores due to high energy requirements for their motility
and feeding mechanisms.
The kinetics of ingestion seem to be fairly linear foi the
first six hours of incubationt thus instantaneous ingestion
rates can be calculated for time periods less than 6 h. Based
on previous work, an incubation period less than six hours
is desirable, because beyond this time the labeled cells will
start to be diluted due to cell division of the predominantly
unlabeled bacteria, thereby reducing the specific activity of
the label. The result is that, the grazer will encounter the
same number of bacteria and fewer of them will be labeled. Also,
considerable recycling of excreted might occur. This will
manifest itself in the data as a decrease in the ingestion rate,
as demonstrated by the radioassay, but in reality the ingestion
rate may be constant through time. On the basis of this informa-
tion, short-term (less than 6 hours) incubations are preferred for
kinetics studies.
In summary, this study suggests that bacterial ingestion
by bactivorous plankton may be at steady state with bacterial
production. A significant amount of ingested bacterial biomass
is used in energy metabolism. The natural plankton population
can be assayed for bactivorous activity under natural concen-
trations and conditions. In order to make more definitive
statements about the role of bacteria in the energy budget of
bactivores, more information about the identity, biomass, and
growth rates of the bactivores is necessary. More attention
must be paid to the species composition and carbon content of
the dominant bactivorous plankton.
SUMMARY
The results presented in this study demonstrate that bac-
teria do serve as a food source for higher trophic organisms
found in Los Angeles Harbor, including Protozoa and some inverte-
brates. These findings support the proposed importance of the
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IIIB 19
269
detrital food web in marine ecosystems currently found in the
literature. It has been suggested that in many ecosystems the
microheterotrophs (bacteria, etc.) play a substantial role in
the nutritional support of higher trophic organisms.
This study suggests that a shift in dominant species to-
wards species that can utilize bactivory for nutrition will be
seen in waters which are organically enriched. There is also
evidence which suggests that bactivory is in steady state with
bacterial production; i.e. bactivore standing stock and feeding
activity will be balanced with bacterial production which, in
effect, is dependent on organic input. For example, areas of
Los Angeles Harbor which receive organic wastes have a compara-
tively high rate of bacterioplankton production and one would
expect high rates of feeding and production among the bactivor-
ous plankters and benthic invertebrates in these waters. From
the evidence presented here and in the literature, one would
expect the microheterotrophs to play a more substantial role
as a food base in these organically enriched systems than in
phytoplankton-based systems. In conclusion, ecosystems which
are bacterially enriched and yet perhaps poor in phytoplankton
production may be as productive overall as other phytoplankton-
based ecosystems.
The study also provides evidence that the assay methods de-
veloped may be employed to investigate bactivory in microscopic
plankton as well as in deposit and filter feeding benthic in-
vertebrates. It is believed that this method, with the appro-
priate modifications, will be useful in elucidating the pathways
and dynamics of carbon in the detrital food web. The major
points of this study are presented below.
1.	Experiments with laboratory isolates of Euplotes sp.
and a marine bacterium were instrumental in developing and dem-
onstrating the accuracy of the techniques. A direct relationship
between Euplotes sp. concentration and rate of removal of bac-
teria from suspension was observed. These data provide evidence
that the results obtained by this method accurately reflect bac-
tivorous activity.
2.	A hyperbolic relationship between grazing rates and
bacterial concentration was suggested. These findings show that
a critical grazer space:bacteria ratio in the experimental design
is necessary to yield maximum potential rates of ingestion, res-
piration, and excretion.
3.	Results from studies with the benthic invertebrates,
Neanthes avenaoeodentata, Macoma nasuta, and Mytilus edulis
indicate that these organisms do ingest bacteria by an assort-
ment of means and utilize bacterial biomass to varying degrees.
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270
IIIB 20
The ingestion rates reported are probably underestimated, because
fecal material was not assayed and because of the reduction in
specific activity due to bacterial cell division during long in-
cubation periods (> 6 h) . It was found that bacterial ingestion
rates varied with experimental conditions.
4. Studies with natural populations of bactivorous plank-
ton {5-20 3 pm) indicate that the rate of removal of bacteria from
suspension by bactivory is directly related to in situ bacterial
concentrations. The results suggest that the grazer's standing
stock, ingestion rates, grazing efficiencies, or a combination
of these will be in equilibrium with bacterial production in nat-
ural planktonic communities.
LITERATURE CITED See Section VI
-------
Linear Regression Correlation
r = 0.996
0.5	1.0	1.5	2.0
Predator Concentration (x 105 ciliates*L~l)
Figure 1. Relationship of the bacterial ingestion rate of the ciliate,
Euplptes sp. to its population density. Data obtained from
laboratory experiments with cultured isolates. Bacterial
concentration was constant in all samples (1.3x10^ cells'L""!)
-------
Ingestion
A Excretion
Respiration
w 25
50
40 ®
Bacterial Density (x
Figure 2. The relationship of bacterial concentration to ingestion, respiration, and
and excretion of bacterial bioraass by the ciliate, Euplotes sp. Data ob-
01
•H
ffl
id
Tl
30 B
u
3
w
20
H
m
O
c»
10
NJ
-J
KJ
H
w
M
M
-------
21 mm Jk.
20—
21 mm
Al 23 mm
>15'
•H
17 mm
GU II IfII
17	mm
18	mm
to
u>
•ri
~ 28 Nov 78
J%r21 mm
*
S
s
18 mm
18 mm
* Numbers -in parentheses represent longest dimension of left valve.
0 18 rm
Incubation Time (h)
w
Figure 3. Metabolically corrected (respiration and excretion) ingestion kinetics of bacteria by
Mytilu8 edulis. A and • represent actual data points and the lines are drawn through the means.
-------
ro
a 4
m
•rf
M
&
-------
30		
50	100	150	200	250	300	350	400
Incubation Time (min)
Figure 5. Gross ingestion kinetics of 14C-labeled bacteria by bactivorous plankton
(5-20 3 pm) from Station A2 in Los Angeles Harbor, 2 Aug 78. Points represent
actual respiration and excretion corrected ingested activity (DPM). The lower
line is a linear regression which doesn't include the 360 min point and the upper
line includes all points.
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276
IIIB 26
B3*
A7
SAN
PEDRO'
B11*
A14.
A3*
AB'
A10*
A13»
AO.
Harbors Environmental Projects
University of Southern California
Figure 6. Stations for microbiological sampling in southern
California coastal and harbor waters. Not depicted are mid-
San Pedro Channel and Santa Catalina Island, Big Fisherman's
Cove station.
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IIIC
277
SEASONAL TRENDS IN TOTAL CHLOROPHYLL a DISTRIBUTION AMONG
SIZE CLASSES OF PARTICLES IN LOS ANGELES HARBOR,
OCTOBER 19 77-DECEMBER 1978
INTRODUCTION
This study was initiated as one part of a more comprehen-
sive investigation of the microbial activity at four stations
in Los Angeles Harbor and one station outside the harbor's
breakwater (Figure 1). Since chlorophyll a remains an often-
used index to the primary productivity and standing stock of a
body of water, any such comprehensive investigation would not
be complete without consideration of chlorophyll a. In this
study, it will be used in conjunction with bacterial standing
stock, measurements of heterotrophic uptake, ATP, and other
parameters.
While seasonal patterns in total chlorophyll a at differ-
ent sites within the harbor and just outside the harbor since
1973 have been described (Oguri, 1974; 1976), this study adds
a new dimension to knowledge about chlorophyll a in the harbor.
The amount of chlorophyll a in particles of six size classes
has been determined monthly at five stations. Knowledge of the
distribution of chlorophyll a among these size classes should
be valuable in correlating the results of the other parts of
this study to the particular groups of phytoplankton.
In an attempt to characterize the nature of these organ-
isms further, floristic analysis was made. For each station
at each month, the species composition and cell numbers were
determined. All of the data available should give a better pic-
ture of the phytoplankton activity of Los Angeles Harbor and
southern California coastal waters than has been possible pre-
viously.
MATERIALS AND METHODS
Samples were collected from the five stations at 1 m depth
in sterile Niskin samplers. Samples were kept in coolers in
darkness until their return to the laboratory. All samples were
filtered within seven hours of their collection.
In all, five different poresize filters (Nuclepore) were
used. Aliquots of the sample were filtered under <5 mm Hg vac-
uum through the following filters: 0.2 pm; 0.6 pim; 1.0 yim; 5.0
pm; 10 pm. In addition, a 37 p screening material (Nitex) was
used in December 1978. Filters were placed in 10 ml of 90%
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278
IIIC 2
acetone and samples were permitted to extract for 24 hours in
darkness and under refrigeration. Before analysis, samples
were allowed to reach room temperature in the dark. Chloro-
phyll a was determined by the fluororaetric method of Yentsch
and Menzel (196 3) and Holm-Hansen, et al. (1965) using a Turner
model 111 fluorometer. The convention of describing a size
fraction, x, is as follows: size fraction x is greater than
0.2 pm but less than 0.6 pm. In symbols this is displayed:
size fraction 0.2pm<^<0-6 pm.
The amount of pigment contained in the following size
classes was determined by difference from total chlorophyll a:
0.2 < x < 0.6; 0.6 < x < 1.0; 1.0 < x < 5.0; and x > 5.0 pm.
Total chlorophyll a was defined as the amount of chlorophyll a
retained by a 0.2 pm poresize filter. In October 1978, an ex-
periment was carried out to determine if, in fact, the amount of
chlorophyll a retained by a 0.2 pm poresize filter truly repre-
sents total chlorophyll a. An Amicon high-presssure, flow
through filter holder was used. A seawater sample from A2 which
had already been filtered through a 0.2 pm poresize filter was
forced through a filter of 10,000 molecular weight retention.
Pressure was supplied by nitrogen gas, and the sample was
stirred by a magnetic stir bar. A 38-fold concentration of the
original sample was attained. Chlorophyll a was extracted by
phase separation and quantified by the fluorometric method.
In August 1978 water samples were collected from 1, 3, 6,
9 and 12 m at station A2. These samples were analyzed for to-
tal chlorophyll a and the distribution of chlorophyll a among
size classes of particles was determined as described above.
In November 1978, this procedure was repeated for a station at
the black buoy west of the reef kelp bed at the Isthmus at San-
ta Catalina Island. At this station, samples were taken from
depths of 1, 10, 20, 30 and 40 m.
Floristic data were collected by Greg Morey-Gaines and
Roseann Ruse. Samples were collected, preserved and stained
with Lugol's iodine solution. Observation and counts were made
with a settling chamber and inverted microscope.
RESULTS
The chlorophyll a data collected are summarized on a month-
ly basis. The data are incorporated in the figures. Unless in-
dicated otherwise, all values represent the average of two rep-
licates. The inherent reproducibility of the fluorometric tech-
nique is about 8% (Kiefer, personal communication), and the av-
erage value for all samples in this study is 5%. In the calcu-
lation of the data for "% of total passing" and "% of total"
chlorophyll a in a given size class, error estimates are pre-
sented as "+ x." These values are not statistical limits, or
standard deviations. They represent the propagation of the
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IIIC 3
279
inherent 8% error in the analytical technique through the
calculations of the values for each column.
Text Table 1 summarizes the results of an experiment con-
ducted on October 4, 19 78 on an A2 sample. The experiment was
designed to determine if, in fact, the chlorophyll a retained
by a 0.2 pm poresize filter real]" represents total chlorophyll
a of a sample. As stated in this table, that pigment which
passes a 0.2 p poresize filter, but is retained by a 10,000
molecular weight filter represents only 0.07% of all pigment
contained in particles of greater than 10,000 molecular weight.
Text Table 1
Results of experiment to determine if the amount of
chlorophyll a retained by a 0.2 pm poresize filter really
represents "total" chlorophyll a of a sample.
1) "Total" chlorophyll a retained by a
0.2 pm poresize filter:
6.46 pg* 1 1
2) Chlorophyll a passing a 0.2 pm filter,
but trapped on 10,000 molecular weight
Amicon filter:
Sample 1
0.0049 jig*1"1
S amp1e 2
0.0037 pg-1-1
3) % of total (6.46 + 0.0043 pg-l~l) pass-
ing a 0.2 )im poresize filter, but retained
by a 10,000 molecular weight filter:
0.07%
Seasonal patterns
Seasonal patterns in total chlorophyll a concentration at
each station are shown in Figure 2. Station AO has a relatively
low chlorophyll a concentration over most of the year, with val-
ues of <4 pg'l~i chlorophyll a. The only feature of note is the
July peak of 15.60 pg-l~l. This burst of chlorophyll a was
short-lived with the values for August through November 1978
near baseline levels.
Station A2 also shows mid-summer concentrations of chloro-
phyll a that are much higher than the rest of the year. The
July 1978 peak was 15.84 }ig*l""l. Evidence for a spring burst
in chlorophyll a exists. The March 19 7 8 value of 6.39 pg-l~l
is well above baseline levels of about 5 pg-l~l. The spring
peak is impossible to define temporally since February and March
data for 1978 are missing, but high levels are maintained from
July through September 1978.
Both the April spring peak and the July 1978 summer peak
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280
IIIC 4
are evident in station A7. In April, the concentration of chlo-
rophyll a reached 9.84 yig-l-1 and in July the value was 22.32
pg*l~l. Note that baseline levels are about 3-5 pg-l~l of chlo-
rophyll a. At station A7 the July peak was short-lived, as at
station AO.
Stations A12 and B9 are extremely similar — not only in
their temporal patterns, but also in the amplitudes of the peak
chlorophyll a concentrations. The sampling of these stations
began in April of 1978. A hint of a spring peak like that seen
in stations A7 and A2 exists. April values drop to about 3
fig*l~l in May and rise slightly in June before rising sharply
in July. The July peak present at AO, A2 and A7 is also pres-
ent at A12 and B9. At A12 the July chlorophyll a concentration
is 21.78 pg'1'1 and at B9 it is 19.08 pg-l~l. Both stations
show a precipitous drop in chlorophyll a at the time of the Au-
gust 1978 sampling. The August value for station A12 was 4.65
pg*l~^ chlorophyll a. That for station B9 was 3.80 pg*l~l. The
September 1978 values nearly equaled those of July — 21.58
pg-l~l for A12 and 18.03 jig*l~^ for station B9. Following Sep-
tember, values at both stations again dropped to lower levels
and show a slight increase from October through December 1978.
Vertical profiles
A vertical profile of chlorophyll a is shown in Figure 3
for station A2 on August 16, 1978. A subsurface chlorophyll a
maximum is indicated at the 3 m depth. The concentration of
chlorophyll a at 3 m is 12.30 pg-l--'-. Chlorophyll a concentra-
tion decreases with increasing depth to a value of 0.466 jag-1"!
at 12 meters.
A similar profile for Isthmus Cove at Catalina Island is
presented in Figure 4. A subsurface chlorophyll a maximum is
indicated at 10 m — a value of 0.68 pg-l-!. Concentration of
chlorophyll a decreases with increasing depth to a value of 0.11
pg*l_1 at 40 m.
Size fractionation
After calculation of the amount of chlorophyll a present
in the various particle size classes, the data were represented
as bar graphs in Figures 5-17. Throughout the first three
months of sampling, October 1977, November 1977 and January of
19 78, the >5 pm size class contained more than 50% of the total
chlorophyll a (Figures 5-7). In April 1978, at stations AO, A2,
A7 and B9, the >5 pm size class only accounted for 22-36% of to-
tal chlorophyll a. The size class of particles between 1 jim
and 5 }im assumed more importance at AO, A2 and A7 with 43%, 56%
and 68% of total chlorophyll a in the >5 pm size class (Figure 8).
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IIIC 5
281
DISCUSSION
The results shown in Text Table 1 demonstrate that the
practice of designating the chlorophyll a retained by a 0.2
jam filter as "total" chlorophyll a is probably valid. In Oc-
tober of 1978, only 0.07% of chlorophyll a contained in parti-
cles of greater than 10,000 molecular weight was contained in
particles less than 0.2 pm. While it is true that this was
done only for one date and one station (A2), it is unlikely
that the results would change by more than one order of magni-
tude. Even so, this would mean that <1% of chlorophyll a is
in the >10,000 molecular weight particles which pass a 0.2 pm
porosity filter.
The seasonal pattern in total chlorophyll a at all stations
show strong peaks in chlorophyll a during the summer. At AO,
A7, A12, and B9, this peak reaches high values in July, and
drops off drastically in August. At station A2, there is a
slight drop off in August, but high values persist until Octo-
ber. Stations A12 and B9 are nearly identical and both show
another peak in chlorophyll a concentration in September of
19 78, with this second peak reaching essentially the same val-
ues as the July peak.
An attempt was made to correlate the amount of chlorophyll
a over time to the number of phytoplankton cells present as de-
termined by direct count methods (Greg Morey-Gaines, personal
communications). However, the floristic data are incomplete,
and any analysis made at this time is to be regarded with cau-
tion. Cell numbers of phytoplankton in March 1978, April 1978,
and May and June 1978 at station A2 were 314/ml, 4,851/ml,
4089/ml and 3,135/ml, respectively. Because these data for
chlorophyll a are missing for March of 1978 at A2, the trends
cannot be considered parallel. In addition, values of chloro-
phyll a dropped from 12.36 pg-l~l to 6.39 jag-l~l between April
and May while cell numbers remained nearly constant. The de-
crease in cell numbers from 4,0 89 per ml in May to 3,135 per ml
in June was accompanied by a decrease in chlorophyll a from
6.39 pg-l~l in May to 4.61 jag*l~l in June 1978.
Another disparity is seen at station A12. In April 1978,
cell numbers were 2,117 per ml, but 2,521/ml in June 1978. The
chlorophyll a values differ greatly: 13.92 pg*l_1 for April
and 3.73 pg*l~l in June.
From these limited data, it would appear that cell numbers
and chlorophyll a probably are not directly correlated. Changes
in species composition could alter the chlorophyll a/cell ratio
and obscure such a relationship.
There are indications in the floristic data of drastic
changes in flora within a month. In March, April and May of
1978, three species of the diatom Chaetoaevos comprised over
-------
282
IIIC 6
80% of the total cell count at all stations. In June of 1978,
flagellates of 5 pm or larger made up at least 44% of the num-
ber of phytoplankton cells. At station AO, this value was 67%.
It is tempting to speculate that the spring peak in chlorophyll
a reported for stations A2, A7 and probably AO was due to the
dominance and presence of these diatoms. It is impossible at
this point to determine if the flagellate-dominated flora was
the reason for the July peak of chlorophyll a found at every
station since floristic data for July aren't available. The
very sharp increase in chlorophyll a is suggestive of very rap-
id increases in cell numbers of a dominant form. Small flag-
ellates, with their rapid rates of cell division are likely
candidates, but this cannot be confirmed.
It should be possible to use size fractionation data to
help determine what type of autotrophs are dominant. However,
most of the data for this first year do not permit such an anal-
ysis. We have already seen that most chlorophyll a is in the
>5 jim size class. However, it is necessary to further subdivide
this class in order to make any decisions about dominant forms
from this type of data. A step in this direction has been made
in adding 10 pm and 37 pm filters to our routine sampling for
chlorophyll a.
From May.through October 1978, the >5 pm size class resumed
importance. During May, no less than 50% of total chlorophyll a
was found in this class (Figure 9). During June, even a greater
percent of total chlorophyll a occurred in this size class —
63%, 70%, 71% and 76% for stations A12, A7, A2 and AO, respec-
tively (Figure 10). July was much the same. From 67-96% of to-
tal chlorophyll a occurred in particles greater than 5 pm (Fig-
ure 11}. Stations inside the harbor had 59-74% of total chlo-
rophyll a in particles >5 pm in August. However, at station AO
- outside the harbor - the total pigment was nearly equally di-
vided among three size classes: >5 pm; 1 < x <5 pm; and 0.2 < x
<0.6 pm (Figure 12). September and October of 1978 demonstrate
this same trend - greater than 4 7% of total chlorophyll a oc-
curred in particles >5 pm (Figures 13 and 14).
In November, a 10 pm poresize filter was added to the array
of filters used in size fractionation. The further resolution
of the >5 pm size class for November is shown in figure 15. Now
we see that the 1 < x <5 pm size class contains the greatest
percentage of the total pigment, in most cases. From 40-51% of
total chlorophyll a was in this size class for stations AO, A2,
A7, A12. At all stations, the percent in the 5 < x <10 pm and
>10 pm size classes was about equal.
In December 1978, even greater resolution was obtained by
adding a 37 pm filtering material to the filtering regime (Fig-
ure 16). The size class of particles between 10 and 37 pm was
most dominant at AO and A2. At other stations, the pigment was
more evenly distributed among the size classes. For this month,
the x > 37 pm size class was not a major contributor to
-------
IIIC 7
283
chlorophyll a.
While the August vertical profile at A2 and the November
vertical profile at Isthmus Cove are not represented with bar
graphs, the distribution of chlorophyll a among the size
classes can be compared. At A2, the pattern for each depth
is much the same as that for each station in August. At 3, 6,
9, and 12 m, >40% of total chlorophyll a was contained in par-
ticles >5 jim. At 1 m depth, only 27% was in this size class.
The November profile is very different. Chlorophyll a seemed
more evenly distributed among the size classes. At 1 m, 58% of
chlorophyll a occurred in particles >10 jam. However, at depths
below 1 m, this value declined. In the range from 10-30 rn depth,
the distribution was quite even. At 40 m depth, there appeared
to be no pigment in particles <1 pm.
Figure 17 shows seasonal changes at a single station, B9.
Floristics
Floristic data demonstrate several notable features. The
spring months of April and May are marked by high numbers of
the diatom Chastoceros soaialis and several other Ckaetoceros
species. Chaetoaeros and the dinoflagellate Gonyaulaz polyedra
were dominant at all five stations in April and at A12 in May
(only A2 data are available for May). In June we found numer-
ical dominance by unidentified flagellates of <5 pm. However,
G. polyedra continued to be very important to total cell carbon
present at harbor stations. Gymnodinium splendens, another
dinoflagellate, was present at A7.
The July samples were dominated numerically, and in terms
of carbon, by a diatom Nitzschia seriata. At AO only did flag-
ellates of <5 pm outnumber N. seriata. In August we see a vari-
ety of species present in the harbor with no real dominance by
any one. At AO, however, the diatom Levtocylindrus danicua was
the most important organism, accounting for >96% of cell number,
volume, and carbon. Inside the harbor, L. danious was present
at A7, A12 and B9. After their nearly total absence in July,
the dinoflagellates G. polyedra and G. splendens did appear at
A2, A7 and A12 in August. However, September was definitely
dominated by dinoflagellates. At all stations, G. polyedra and
G. splendens were the most important phytoplankters present.
November appears to be a transition time for the flora.
Some G. polyedra is still important at AO and A2. However, the
small flagellates have reappeared in greater numbers - in fact,
they are the most abundant phytoplankter at all stations. Levto-
cylindrus daniaus is important at all harbor stations, but not
at AO. AO has the dinof lagellate Ceratium furoa as an important
contribution to volume and carbon.
The December 19 78 flora resemble no other month's flora.
-------
284
IIIC 8
Chaetoosvos species and Ceratium species are most important. A2
and A 7 have large numbers of the diatom Rhizosolenia.
Another apparent switchover occurs in January 1979. These
samples were dominated both numerically, and to a great extent,
in carbon by small, unidentified flagellates.
An attempt was made to correlate the chlorophyll a over
time with cell number. The coefficient of determination, r2,
was only 0.40. Attempts to correlate chlorophyll a concentra-
tion with cell volume and cell carbon as estimated from cell
volume, resulted in about the same degree of correlation. While
poor correlation with cell numbers is not a surprise, the reason
for poor correlation with cell carbon is not clear. Perhaps
the inherent assumption that cell carbon:chlorophyll a ratios
are constant is not a good one.
The seasonal trends demonstrated in 1978 in chlorophyll a
do agree well with those reported by Oguri (19 74; 1976) for Los
Angeles Harbor. The spring and summer peaks seem characteris-
tic, both in timing and in relative amplitude.
At most months, the small scale resolution of our filtering
array does not permit much correlation between species present
and the size classes of chlorophyll a. Most organisms presently
identified are very much larger than 5 pm, as reported in the
results section, >50% of total chlorophyll a is usually found in
the particles >5 pm. However, in November of 1978, the further
resolution provided by the addition of a 10 pm poresize filter
does enable some interpretation. In November, flagellates of
<5 pm were important numerically, and in terras of cell carbon.
This is reflected in figure 15, which shows that for most sta-
tions, the size class of particles between 1 and 5 pm contained
a greater percentage of chlorophyll a than any other size class.
In December 1978, flagellates virtually disappeared. Figure 16
demonstrates the relative unimportance of the 1 < x <5 pm size
classes during December. The presence of larger diatoms (Chae-
tocsros) and dinoflagellates (Ceratium) is reflected in the
greater importance of the >10 pm size classes. With continued
use of these larger filters, our ability to correlate the flora
with the distribution of chlorophyll a among size classes of
particles should improve.
Station AO consistently had lower values of chlorophyll a
concentration than the stations within the harbor. This is not
surprising, and in fact, is to be expected. Likewise, the val-
ues for chlorophyll a at 1 m at Isthmus Cove, 0.43 pg-1"1, in
November 1978 are much lower than 1 m at AO in November 1978
(0.98 pg*l~l). Thus, a distance-offshore dilution effect is
seen. All stations within the harbor maintained about the same
levels of chlorophyll a. Stations A12 and B9 are extremely
similar in their seasonal pattern and amplitudes - probably be-
cause of their physical proximity. A7 demonstrated the highest
-------
IIIC 9
285
levels of chlorophyll a. The fact that the same seasonal pat-
terns are found at station AO as at the harbor stations demon-
strates that the causative conditions are not unique to the har-
bor. Conditions in the harbor may enhance the magnitude of the
effect.
-------
286
IIIC 10
SUMMARY
The seasonal trends in chlorophyll a concentration for
phytoplankton of Los Angeles Harbor and adjacent coastal water
in 1978 are reported. These data show persistence, in 1978,
of patterns reported for harbor waters since 1972 (Oguri, 1974;
1976). The spring peak occurs in April for stations within
the harbor. At the one station outside Los Angeles Harbor, the
April peak is much less pronounced. The major peak occurred in
July when chlorophyll a concentrations reached values of >16
pg'l-1 for all stations. This July peak was associated with a
bloom of the diatom Nitzsohia seviata. Chlorophyll a concen-
trations at stations within Los Angeles Harbor were always
greater than those at station AO, outside the harbor. Two sta-
tions, A12 and B9, are extremely similar - not only in the
trends of chlorophyll a, but also in the magnitude of chloro-
phyll a concentrations. This is probably a reflection of their
physical proximity (Figure 1).
Fractionation of chlorophyll a into the following size
classes: 0.2 < x <0.6 pm; 0.6 < x <1 pm; 1 < x <5 pm; and >5 pm
for October 1977-October 1978, demonstrated that >50% of the
chlorophyll a was contained in particles >5 pm. In November
19 78 further fractionation demonstrated that greater than half
of this amount was, in fact, contained in particles >10 pm.
With the addition of a 37 pm poresize filter in December of
1978, even further subdivision was made. Between 15% and 30%
of total chlorophyll a was found in the size class >37 pm for
this month.
LITERATURE CITED: See Section VI
-------
IIIC 11	287
B6j
C11«
WILMINGTON
LONG BEACH
B5.
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o •
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Lo
A14.
A8
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A13«
B1«
A0«
SCtli III HUES
Harbors Environmental Projects
University of Southern California
Figure 1. Station locations for chlorophyll a and size
fractionation studies.
-------
288
IIIC 12
Figure 2. Seasonal changes in chlorophyll a at stations
AO, A2, A7, A12, and B9.
-------
IIIC 13
289
Chlorophyll a pg-1
Figure 3. Vertical profile of total chlorophyll a con-
centration at station A2 on August 16, 1978.
-------
290	IIIC 14
Chlorophyll a >ig-l~^
0.1 0.2 0.3 0.4 0„5 0.6 0o7 0.8 0.9 1.0
_l	I	I	1	I	I	I I I	L.
10m
to

Q
30m
40m
50ra
Figure 4.
Vertical profile of chlorophyll
in Isthmus Cove on November 17,
a concentration
1978.
-------
Oct% 10, 1917
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-------
Nuv. ?. J 97 V
AO
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A2
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PARTICIiaTE SIZE CUSSES
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Figure 7. Distribution of chlorophyll a among size classes for January 11, 1978 samples
-------
April 5, 19?8
AO
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PARTICULATE SIZE CUSSES
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-------
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Figure 13. Distribution of chlorophyll
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Dec. 6, 1978
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Figure 16. Distribution of chlorophyll a among size classes for December 6, 1978 sample.
-------
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Figure 17. Distribution of chlorophyll a among size classes for April 5, 1978, July 5, 1978,
September 6, 1978, October 4, 1978, November 2, 1978, and December 6, 1978 samples.
UJ
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| Reproduced from
-------
Intentionally Blank Page
-------
HID
305
THE UPTAKE, SIZE FRACTIONATION, AND TURNOVER TIME
OF ORTHOPHOSPHATE BY BACTERIOPLANKTON AND PHYTOPLANKTON
IN THE LOS ANGELES HARBOR AND COASTAL WATERS
INTRODUCTION
Orthophosphate is one of several inorganic nutrients
studied in characterizing the microbial activity in the outer
Los Angeles Harbor. This nutrient is universally required by
organisms and may be assimilated directly as PO<,~2 only by
bacterioplankton and phytoplankton. Oceanic concentrations of
phosphate are often at the lower limits of detection (0.03 to
Symole*liter"1) while turnover times, especially during phyto-
plankton blooms, are very short, usually minutes (Campbell, 1977).
The purposes of this investigation are fourfold:
1.	To compare rates of uptake of orthophosphate, in situ
phosphate concentrations, and turnover times (Tt) for
phosphate between a station (A2) located inside the
eutrophic Los Angeles Harbor and one (AO) outside the
harbor breakwater, and among several depths at
station A2.
2.	To determine which size fractions of microplankton
are responsible for the assimilation of orthophosphate.
3.	To describe seasonal changes in turnover times for
phosphate, especially in correlation with standing
stocks of bacterioplankton and phytoplankton, and
with the in situ phosphate concentration.
4.	To evaluate the role of bacterioplankton in nutrient
cycling in the food web of the outer harbor.
METHODS
In July, August and September 1978, samples of sea water
were collected with sterile Niskin samplers from 1 m below the
surface at four stations inside the harbor (A2, A7, A12, B9)
and one station (AO) outside the harbor breakwater (see Figure
11) and filtered through a 203 jam Nitex net to remove larger
plankton. Aliquots of 50 or 100 ml of each sample were fil-
tered through a 0.2 pm pore-sized filter (Nuclepore) and
stored frozen before dissolved reactive phosphate determination
by the spectropftotometric method of Strickland and Parsons (1972) .
All filtrations employed discrete pore-sized membrane
filters (Nuclepore) which are hereafter designated as x pm
filters where x = 0.2, 0.6, 1.0 or 5.0 pm. For the studies
Preceding page blank
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306
HID 2
on uptake kinetics a 300 ml sample was incubated with stirring
at 18 C and 3000 lux with approximately 1.3 yci of either
carrier-free Hb32PO<» or H33 PCH . The addition of carrier-free
label does not significantly alter the ambient phosphate con-
centration. At various intervals for up to 6 hours, 10 ml
subsamples were filtered in duplicate through 1.0 Um and 0.2
ym filters at -10 cm Hg pressure, rinsed twice with 5 ml
prefiltered 0.2 ym OC sea water (SW) and dried. The dried
filters were placed in 10 ml of a toluene-based scintillation
co.cktail for counting in an LS-100 Beckman counting system.
A 1 ml aliquot of the unfiltered sample was counted directly
in Aquasol to yield total disintegrations per minute (dpm)
of the radioisotope per volume of sample. Counts per minute
(cpm) were converted to dpm by means of a quench curve.
Uptake values were corrected for nonbiological adsorption
of the label by subtracting either an acid-killed or zero-hour
(t0) blank for each time point. Acid-killed controls were
prepared by addition of 0.2 ml of 7% PCA to 100 ml of sample
immediately before addition of the label; the final pH was 2.
TQ controls were prepared by filtration of duplicate 10 ml
subsamples immediately after addition of label to a live
sample.
For the size fractionation studies, 200 ml seawater
samples were incubated for 24 h under the conditions described
above. At the end of this period, duplicate 10 ml subsamples
were filtered onto 5.0, 1.0, 0.6 and 0.2 vim filters, which
were rinsed, dried and counted as above. These filtrations
were nonsequential. Either acid-killed or tQ controls were
prepared for each sample.
Specific activity of the radio label was calculated from
the dissolved reactive phosphate concentration and the total
dpm ml-1 of unfiltered sample. The particulate uptake was
calculated by conversion of dpm ml-1 to nmole POi»"2•liter"1,
using the specific activity in nmole POi,"2 *dpm~1 . Uptake was
plotted against incubation time to obtain a line, the slope
of which was determined by linear regression analysis and
expresses the particulate uptake rate, in nmole P0i»~2-liter-1
•h-1- Turnover times (Tt), defined by the equation
where s = natural phosphate concentration in ymole POi* •
liter-1 and v = particulate uptake rate, were calculated both
from the kinetic experiments and from the single point incuba-
tions where uptake was size-fractionated. The percent uptake
passing each filter size was computed, based on 200% reten-
tion by the 0.2 ym filter.
Specific uptake rates, reported as nmole POt,-2 •mgC-1 -h~1
are based on the assumption that uptake by the size fraction
>0.2 ym, <1.0 ym is due to bacteria, while uptake by the size
class >1.0 ym is primarily due to phytoplankton. This assump-
tion is based on our own observations and on the data of Faust
-------
HID 3
307
and Correll (1976) and Harrison et al. (1977), which showed
that at least 80% of phosphate uptake by cells >1.0 urn is
algal, while at least 90% of the uptake passing a 1 ym filter
and retained by an 0.2 ym filter is bacterial. Determination
of bacterial and phytoplankton standing stocks was by the
acridine orange direct counting technique (AODC) according to
Daley and Hobbie (1978) and chlorophyll a biomass estimates by
the fluorometric technique of Strickland and Parsons (1972).
These values were converted to mgC as explained elsewhere in
this report.
RESULTS
The kinetics of phosphate uptake were studied for three
size classes of microorganisms at stations A2 and AO (Figures
1 and 2). Total uptake and uptake by the >1.0 ym size class
were measured directly; uptake by the bacterioplankton was
determined by subtraction. In all cases uptake was linear over
6 hours, with a correlation coefficient >0.93 at a significance
level of 0.05 when analyzed by least squares linear regression.
At station A2, where the reactive phosphate concentration
was measured at 0.59 ymole PCH"2-liter"', turnover time of
phosphate was 41.2h and uptake rate (v) was 14.4 nmole PCU~2•
liter" ^h-1 for the total population. For the >1.0 ym size
fraction, T^ was 74.1 hours, and v was 8.0 nmole PO2'liter"1•
h~1, while Tt and v by the bacterioplankton were 95.6h and
6.8 nmole POi»~2 'liter" 1 -h- 1 respectively.
At station AO, where the phosphate concentration was
0.78 ymole P(K~2-liter"1, v for the total population was
slower, 11.18 nmole PCU"2-liter" 1 *h~1 and Tt was 1.5 times
longer, at 69.6 hours. The bacterioplankton population took
phosphate up at a rate similar to that at A2, with a
v = 7.1 nmole PO<»~2-liter"1*h~ 1 and Tt of 109.8h. Bacterio-
plankton uptake at AO was 50% of the total, while at A2 it
was 30% of the total. The bacterioplankton standing stock
at A2 was 5-fold that at AO for this sampling date.
Figures 3 and 4 present the data for the vertical profile
study of phosphate uptake at station A2. Only total uptake was
measured. Although the in situ phosphate concentration dif-
fered by less than 0.35 ymole POi, 2 -liter-1 in the upper
10 meters, T^ increased greatly with depth below 3 meters.
The sample collected at 3m showed the highest uptake rate, at
8.74 nmole P(H~1 • liter"1 -h"1 with a Tt of 139 h (5.8 days).
The sample collected from 9 meters showed no significant
uptake after 3 h. Figure 4 indicates that uptake rate closely
parallels the natural phosphate concentration with depth.
Figure 5 presents the data for an experiment, in which
the total uptake at station AO was compared to that at station
-------
308
HID 4
A2, over a 5 h incubation period. Only total uptake was
measured, which was linear for both stations with a correla-
tion >0.95 at the 0.05 significance level. At station A2,
the uptake rate for this experiment in September was more
than triple that of the first two experiments done in July
and August, with a September value of 49.1 nmole PCK~2'liter"1
.h~ 1. The was 25.5 h and the phosphate concentration was
similar to that measured in August: 1.25 ymole PO^" -liter-
and a 1.12 ymole PO2 -liter"1, respectively. At AO the
uptake kinetics for September approximate those in July, the
being 75.1 h and the uptake velocity = 10.4 nmole PO »~"2 •
liter" l,h-1. The phosphate concentration was the same in both
months, 0.7 8 ymole PO*-2-liter-1.
Table 1 summarizes all of the kinetic data. In Figures
6-10 and Tables 2-4, the results of the 24 h fractionation
studies are given.
At all stations but A7, the uptake rate at least doubled
in September as compared with July. At A7 the uptake rate
actually decreased in September. This sample may have become
anoxic during the long incubation, as it smelled strongly of
H2S when the 24 h filtrations were performed. Aside from
this anomalous rate decrease at station A7, several patterns
emerged from the data: 1) In both months, AO had the lowest
rates of uptake by all size fractions; at this station the
microorganisms <5 ym in smallest diameter were responsible
for at least 80% of the total uptake, while those <0.6 ym
achieved at least 55% of the total uptake; 2) at stations A12
and B9 similar rates of uptake were obtained for a given
sampling period. At these stations about 70% of the total
uptake passed a 5 ym filter, 55% passed a 1 ym filter, and
50% passed a 0.6 ym filter; 3) although the uptake rate at
A2 is more than doubled from July to August, the uptake dis-
tribution was about the same for both months, with about 80%
passing a 5 ym filter, 70% passing a 1 ym filter, and 55%
passing a 0.6 ym filter; 4) a dramatic increase in the percent
uptake retained by the 0.6 ym and larger pore-sized filters
was seen in the September samples of stations A2 and A7. At
station A2, 56% of the total uptake passed a 5 ym filter and
25% passed a 0.6 ym filter in September, whereas these values
were 76% and 54%, respectively, in July. At A7, 15% of the
total uptake passed a 5 ym filter and 7% passed a 0.6 ym
filter in September, as compared with 80% and 38%, respectively,
in July.
Table 5 (D. Krempin, personal communication) shows the
bacterioplankton standing stocks at these five stations for
the two months sampled. At every station the population
increased at least twofold in September. A dinoflagellate
bloom of Gymnodinium splendens also occurred in the Los Angeles
Harbor in September. Phytoplankton standing stocks, estimated
from yg chlorophyll a liter-1 are found in Table 6 (J. S00H00,
personal communication). In Table 7, orthophosphate uptake
-------
HID 5
309
per yg bacterioplankton and phytoplankton carbon has been
calculated. Phosphate uptake by bacterioplankton per unit
biomass proceeds at a rate 17 to 1000 times greater than that
by phytoplankton. However, because the phytoplankton stand-
ing stock is 60 to 400 times bacterioplankton biomass, the
rates at which these two size classes take up phosphate are
of approximately the same magnitude on a volume basis.
In Figures 6 through 10 and Tables 2 through 4, the
results of the 24 h fractionation studies are presented. At
all stations but A7 uptake more than doubled in September
compared with July. In July, bacterioplankton are responsible
for at least 50% of total uptake at all stations, while in
September, uptake by the >1.0 vim size fraction was over 60%
of the total at stations A2 and A7. Turnover times of phos-
phate by the total population ranged from 47 h at station B9
in September to 159 h at station AO in July.
DISCUSSION
Although phosphate is a limiting nutrient in many oceanic
environments, this is probably not the case in the eutrophic
waters of the Los Angeles Harbor. The range of concentrations
for the five stations discussed in this report was 0.5 to 3.0
ymole POi»~2 • liter-1 over the three months in which phosphate
was measured. This is similar to concentrations found in the
coastal region around San Diego: 0.64 to 2.34 ymole PO., "z • liter-1
(Solorzano and Strickland, 1968) and in the Rhode River sub-
estuary of Chesapeake Bay: 4.0 ymole PC\ ~2•liter-1 (Frieble
et al., 1978). It is high compared to that measured off
La Jolla, California: 0.2 to 0.7 ymole PCK ~2* liter-1 and in
the oligotrophic waters of the East-Central Pacific; 0.05-0.70
ymole PO<* ~2 • liter"1 (Solorzano and Strickland, 1968).
Two important points emerge from the kinetic data (Table 2):
1. At station A2, a doubling in the bacterial popula-
tion (Table 5) from July to September, 1978 corre-
sponded with a threefold increase in uptake rate.
Phosphate concentration also doubled over this
interval, whereas phytoplankton biomass decreased
slightly (Tables 5, 6). From July to August the
bacterial population at station A2 dropped 30% and
uptake rate was correspondingly halved; in situ
phosphate concentration almost doubled. Phyto-
plankton biomass also dropped about 30% in August.
These changes were correlated with TITP in August,
following chlorination during a breakdown of the
treatment system from May through July. These
observations are in agreement with the finding of
Faust and Correll (1976), that the phosphate assimi-
lation ability of bacteria, and algae in the Chesa-
peake Bay had high correlations only with biomass,
-------
310
HID 6
and were not influenced by the in situ phosphate
concentration.
The data for station AO are more difficult to
interpret. Neither phosphate concentration nor uptake
rate changed significantly between July and September
despite a tripling of the bacterial population.
Phytoplankton decreased more than sevenfold, but the
data presented in Figure 1 indicates that uptake by
this size class is less than 50% of the total. While
the specific uptake rate (nmole-109 cell-1**!""1) at
A2 varies only about twofold, from 6.4 in July, to
12.0 in September, at AO it ranges fourfold, from 7.0
in September to 28.7 in July. In January (unpublished
data) this rate drops to 2.2 at AO. This indicates
that the bacterioplankton at A2 are metabolically more
active and less variable than those outside the
breakwater.
2. The results of the vertical profile (Figure 4) show
a subsurface maximum for bacterial and phytoplankton
density as well as for the in situ phosphate concen-
tration and the uptake rate by the microplankton at
a 3 meter depth, followed by a uniform decrease in all
these parameters to 0 m depth. This differs from the
depth profiles of Harrison et al. (1977) off British
Columbia, which showed maxima for both phosphate
assimilation and in situ concentrations below 10 m
depth.
A phytoplankton bloom occurred throughout the harbor in
July (Table 6) but the standing stock dropped in August. In
September, phytoplankton peaks equal in magnitude to the July
bloom occurred at stations A12 and B9, while bacterial popula-
tions increased two- and 1.3-fold, respectively. Relative
uptake into different size classes did not change significantly
(Figures 8 and 9). Smaller phytoplankton blooms were measured
at stations A2 and AO in September, when bacterial populations
increased 1.8- and 4-fold, respectively, over the July stand-
ing stocks. Table 7 shows that the uptake of phosphate per
yg phytoplankton carbon increased at every station from July
to September, but the increase was greatest at station AO
(18-fold) and A2 (6-fold). The uptake of phosphate per ug
bacterial carbon remained about the same for all stations
except A7, where it decreased 28-fold between July and Septem-
ber. The uptake data for station A7 may be an artifact of
that sample having become anoxic, as discussed earlier. The
trends at the other stations, however, suggest a hypothesis:
early in a phytoplankton bloom (such as at stations AO and A2
in September) the phytoplankton population may "gear up" for
rapid growth and divisions by storing up nutrients, such as
phosphate, beyond their immediate needs. The physiological
status characterizing these organisms at that time would
-------
HID 7
311
enable them to compete better with bacterioplankton for dis-
solved phosphate. Later in the bloom (as in the July peak),
the phytoplankton probably play a less important role in
phosphate uptake, relative to the heterotrophic population.
The bacterioplankton are increasing in response to the greater
availability of dissolved organic carbon in the water column,
presumably resulting from phytoplankton excretion lysis and
grazing effects. Since the uptake rate per unit of bacterio-
plankton biomass exceeds that per unit of phytoplankton
biomass by 17- to 1000-fold, the greater density of bacterio-
plankton enables these heterotrophs to outcompete the phyto-
plankters for nutrients.
This hypothesis is not consistent with the observation
of Faust and Correll (1976) that higher phosphate assimila-
tion by algae is due to higher numbers of algae rather than
higher phosphate-assimilation ability per cell, but is
supported by laboratory culture experiments of Rhee (1973)
and Lean and Nalewajko (1976), which demonstrated the ability
of algae to store excess phosphorus before undergoing
several cell divisions.
Turnover times reported here (Tables 1, 2, 3 and 4) are
high. Campbell (1977) states that turnover times are meas-
ured in minutes at the sea surface during summer months.
However, the shortest turnover time recorded in the present
study was 25 h at station A2 in July. This is undoubtedly
the result of a high phosphate input into these coastal
waters and/or rapid recycling of the resident POt*-2 in the
water column, since the phosphate concentration remained high
even during the phytoplankton blooms. The overall predomin-
ance of bacterioplankton in phosphate uptake (Table 7)
supports the findings of Fuhs et al. (1972) and Rhee (1972)
that phytoplankters are out-competed by bacteria when the
phosphate concentration is greater than 0.1 umole PCK ~2•liter-1.
However, the long turnover times recorded here suggest that
POi,~2 is not a limiting nutrient to phytoplankton in the Los
Angeles Harbor. This conclusion is supported by evidence
presented elsewhere in this report, where laboratory cultures
of phytoplankters were grown in harbor sea water supplemented
with various concentrations of TITP effluent.
CONCLUSIONS
The following points summarize the experimental results
concerning the uptake of orthophosphate by the bacterio-
plankton and phytoplankton in the Los Angeles Harbor and
Coastal waters.
1. In situ concentrations of POu"2 at four stations (A2, A7,
A12, B9) inside the harbor and one (AO) outside the
harbor breakwater are found within the range 0.5 to 3.0
-------
312
HID 8
pmole-liter-1 over the period from July 1978 to February
1979. These values are consistent with measurements
reported in the literature for highly eutrophic waters.
2.	Uptake of orthophosphate (as H3 3 2 P04 and H333P04) is
linear over seven hours by the >0.2 ym microplankton in
seawater samples from a 1 meter depth at stations A2
and AO. At AO, uptake rate per volume water is remark-
ably constant, ranging from 10.4 to 11.2 nmole POi»~2 • liter-1
. h_1 between July and January. At A2, the uptake rate
varies 7-fold, from 49.1 to 6.7 nmole PO4 ~2 • liter-1 -h-1
in September and January respectively. By contrast, the
specific uptake rate (nmole POif-2»109 cell_1*h-1) varies
13-fold at AO, from 2.2 in January to 28.7 in July,
whereas at A2 the specific uptake rate varies only 2.5-
fold. This indicates that the bacterioplankton at A2 are
metabolically more active and less variable than those
outside the breakwater.
3.	A vertical profile at station A2 in August revealed sub-
surface maxima both for POt" concentration and phosphate
uptake rate by the microplankton at 3 meters, the depth
at which the greatest bacterioplankton and phytoplankton
population densities also occurred.
4.	The shortest turnover time measured was 25 hours at A2 in
September 1978. The mean turnover time at AO was 64 hours.
These data suggest that P04~2 is not a limiting nutrient
in these highly eutrophic waters, a conclusion also
reached with laboratory cultures grown in harbor sea water
supplemented with various concentrations of TITP effluent,
as reported elsewhere in this volume.
5.	Fractionation of the uptake data into four size classes
(>5.0, >1.0, >0.6, >0.2 ym) showed that, while generally
60 to 80% of the label passed a 1.0 um filter and can be
considered with bacterioplankton, up to 60% of the label
was retained by a 1.0 ym filter in September, and 50% in
July. This increase in uptake by the larger size class
is strongly correlated with the occurrence of phyto-
plankton blooms in these months. Overall, the uptake rate
per unit bacterioplankton carbon was 17 to 1000 times the
rate per phytoplankton carbon. The difference in uptake
rates was much greater in July, when phytoplankton
biomass was 60 to 400 times bacterioplankton biomass,
than in September, when the biomass difference was 8- to
55-fold. These data indicate that bacterioplankton are
the predominant organisms involved in uptake of dis-
solved orthophosphate in the Los Angeles Harbor.
LITERATURE CITED: See Section VI.
-------
Table 1. A summary of kinetic data for orthophosphate uptake at stations A2 and AQ, Summer 1978
Sample date
Sample depth
meters
Station
Size fraction
Involved
s
-7 -1
ymole PO^ -liter
V
nmole PO^-llter"1-!)"^
Tt hours
7-5-76
1 m
A0


11.18
69.6


>0.2um,1.Oym
0.78
7.10
4.13
109.8
190.8
7-5-78
1 m
A2
>0.211111

14.37
41.2



>0.2ym,1.Oum

7.99
74.1
8-16-78
1 m
A2
>0.2pm
1.12
7.91
141.6

3 m

H
1.22
8.74
139.6

6 m

II
0.97
4.26
227.7

9 m

l<
0.88
0.35
2521.5

10 m

II
0.96
2.87
334.5
9-6-78
1 ro
A0
>0. 2|jm
0.78
10.39
75.1

1 ra
A2
>0.2ym
1.25
49.12
25.5
u>
h-»
U>
-------
Table 2. A summary of the 24 hour size fractionation data for orthoPhos^^|1u^^^d7^3lpo4
Stations: AQ	A?	A^	Bg
Size Fraction
{(fill)
dpm + s D
retained ' '
dpm ± s 0
retained xu'
dPm ± S D
retained i-u-
^P"1 + c n
retained ' '
dP™ ± S D
retained a*u*
6
2515 * 89.0
10321 ± 835
8431 ± 26.2
11292 ± 268
16380 i 326
1
7179 ± 34.5
11592 ± 439
17060 ± 72.7
19528 ± 178
22059 ± 993
0.6
4015 ± 164
19843 ± 665
24397 ± 284
22114 ± 181
25123 ± 404
0.2
13197 ± 422
42986 ± 1138
39672 ± 1221
41322 i 1965
47210 ± 3050

nmoles retained
•L-V1
nmoles retained
•L-V1
nmoles retained
i"1 h-1
•L -h
nmoles retained
i-l i-l
•L *h
nmoles retained
•L-V1
5
0.929 ± 0.062
2.25 ± 0.18
11.4 ± 0.03
2.68 ± 0.14
2.45 ± 0.02
1
1.285 ± 0.013
2.53 ± 0.09
23.2 ± 0.09
4.51 ± 0.04
3.40 ± 0.09
0.6
1.572 ± 0.065
4.35 ± 0.14
33.2 i 0.38
5.11 ± 0.04
3.88 ± 0.07
0.?
4.876 ± 0.156
9.44 i 0.25
54.1 ± 1.66
9.55 ± 0.45
7.68 ± 0.08

% passing
% passing
t passing
X passing
% passing
5
80.9
76.1
78.7
71.8
68.0
1
73.6
73.1
57.0
52.7
55.6
0.6
67.7
53.8
38.5
46.5
49.4
0.2
0
0
0
0
0

Tt hours
Tj. hours
hours
Tj. hours
hours
5
837
262
259
178
162
1
605
233
128
106
117
0.6
494
135
89
94
103
0.2
159
62
55
50
52
ijmole P0^-liter '
0.78
0.59
2.98
0.48
0.40
-------
Table 3. A summary of the 24 hour size fractionation data of orthophosphate uptake foe 5 depths
at station A2, 8-16-78. Label assayed
	Uepth 	lm		3m	6 nn	9 m	 	XO m	
Size Fraction
(nm)
df"" + s D
retained ~ ' "
dPm ± S D
retained 5-u-
, ~ S D
retained
dpm +so
retained ~ ' *
dpm + s D
retained "
5
36S2 ± 123
4383 ±84.7
4024 ±213
3320 ± 288
5462 ± 431
1
9G86 ± 293
12640 ±24.1
8323 ±57.7
5923 ± 10
12212 i 46.1
0.6
12245 ± 436
16161 ± 157
11093 ± 354
6683 ± 127
17628 ± 107
0.2
31073 ± 443
39995 ± 493
22161 ± 4285
11871 ± 3321
35287 ± 438

nmoles retained
•L-V1
nmoles retained
•L-V
nmoles retained
i-' k"1
•L -h
nmoles retained
•L-V1
nmoles retained
•L-V1
5
2.8 i 0.09
3.36 ± 0.06
2.63 ± 0.13
2.02 ± 0.17
3.66 ± 0.29
1
7.42 ± 0.22
9.69 ± 0.01
5.44 ± 0.03
3.60 ± 0.00
8.19 ± 0.03
0.6
9.38 + 0.330
12.39 ± 0.06
7.25 ± 0.23
4.09 ± 0.10
11.82 + 0.07
0.2
23.82 ± 0.34
30.660± 0.37
14.49 t 2.80
7.22 ± 2.02
23.67 + 0.29

% passing
X passing
% passing
% passing
X passing
5
83.2
89.2
81.8
72.0
84.50
1
68.8
68.4
62.4
50.1
65.5
0.6
60.5
59.5
50.0
43.3
50.0
0.2
0
0
0
0
0

hours
hours
Tt hours
Tj. hours
hours
5
400
363
368
435
261
1
150
125
178
244
117
0.6
119
98
133
215
81
0.2
47
39
64
121
40
Ijrnole PO^-liter"'
1.12
1.22
0.97
0.88
0.96
OJ
M
Ul
-------
Table 4. A summary of the 24 hour sizo fractionation data for orthophosphate uptake, 9-6-78.
Label assayed Hj^PO,)
Stations: Ag	A^	A^
Size Fraction
(pro)
??m , ± S.D.
retained
^ . ± S.D.
retained
fP? ± S.O.
retained
dpra + s 0
retained ~ ' "
dpm + s D
retained ~ ' '
5
3576 ± 677
13342 ± 600
9841 i 679
9425 ± 126
9309 ± 133
1
6749 ± 5B4
19033 t 81.4
9601 ± 222
11918 i 1211
14152 t 401
0.6
13847 ± 218
23038 ± 450
10722 ± 162
15233 ± 2007
17935 ± 1088
0.2
31655 ± 292
30880 ± 344
11606 ± 753
24361 ± 1455
35395 ± 1977

ninoles retained
•L-V1
ninoles retained
•l/V1
nmoles retained
•L"1-h"'
ninoles retained
i"1 h"l
•L *h
ranoles retained
•l/V1
5
1.59 ± 0.30
8.26 ± 0.37
8.24 t 0.56
7.81 ± 0.10
4.61 ± 0.12
1
2.99 ± 0.26
11.67 ± 0.05
8.10 ± 0.18
9.88 ± 2.17
7.01 ± 0.13
O.C
6.15 t 0.09
14.19 t 0.28
8.98 ± 0.13
12.63 ± 1.66
8.89 ± 0.54
0.2
14.08 ± 0.13
19.06 ± 0.21
9.72 ± 0.63
20.20 ± 1.20
17.55 ± 0.98

X passing
% passing
% passing
X passing
% passing
5
88.7
56.6
15.2
61.3
73.7
1
78.7
38.7
16.5
51.0
60.0
0.6
56.3
25.5
7.6
37.4
49.3
0.2






hours
Tt hours
Tt hours
Tt hours
Tt hours
S
490
140
173
185
182
1
260
107
176
146
119
0.6
126
88
159
114
94
0.2
55
65
147
71
47
pinole PO^-llter"1
0.78
1.25
1.43
1.45
0.84
-------
Table 5.
Standing stocks and biomass estimates of bacterioplankton at 5 stations in
tlio Los Aiiyeies Harbor area, July - September 1978. Estimates are based on
direct counts tisiny epifluorescent microscopy.
Sample date
Station
, Sample depth
(meters)
Total bacterioplankton
<-105)-ml_1
Bacterial biomass
lig C-liter '
7-5-78
*0
1
m
3.9
t
0.6
3.1
±
0.5

*2
1
m
22.5
±
2.7
17.6
+
2.1

*7
1
m
29.1
±
2.3
22.8
+
1.8

*12
1
m
16.9
±
0.9
13.2
±
0.7

B9
1
m
27.6
±
2.1
21.6
±
1.6
8-16-78
A2
1
m
11.4
±
3.6
12.8
+
2.8


3
m
18.9
±
3.6
14.8
±
2.8


6
m
15.7
±
2.4
12.3
+
1.9


9
m
10.5
+
3.1
8.2
±
2.4


10
m
9.2
±
3.2
7.2
±
2.5
9-6-78
*0
1
m
14.8
±
1.9
11.6
±
1.5

A2
1
m
40.8
±
4.0
31.9
±
3.1

A7
1
m
43.3
±
5.6
33.9 ±
4.4

*12
1
m
36.2
±
6.2
28.3
±
4.9

09
1
m
35.4
±
4.4
27.7
±
3.4
u>
M
-------
318
HID 14
Table 6. Standing stock estimates of phytoplankton biomass at
5 stations in the Los Angeles Harbor, July - September
1978. Estimates are based on chlorophyll a measurements
according to the method of Strickland and Parsons, 1972.
Sample date
Station
yg chlorophyll a/liter""'
mg phytoplankton C-liter"^
7-5-78
Ao
15,5
1.2
*
*2
15.9
1.2

A7
42.2
1.3

A12

1.65

B9
22.0
1.5
8-2-78
Ao
0.1
0.1

a2
10.5
0.7

*7
8.0
0.7

A12

0.4

B9
4.8
0.3
9-6-78
A0
1.0
0.15

a2
10.8
0.9

*7
4.1
0.3

A12

1.6

B9
22.0
1.35
-------
HID 15
319
Table 7. Relative uptake of orthophosphate by bacterioplankton
and phytoplankton based on biomass estimates of the
standing stocks, summer 1978.
Sample date
Station
Uptake rate of
bacterioplankton
nmole P0^*yg
Uptake rate of
phytoplankton
,-2
-1 u-1
nmole PCL «yg C -h
7-5-78
12
1.16
0.392
1.35
0.379
0.199
0.001
0.002
0.018
0.003
0.002
9-6-78
12
0.957
0.232
0.047
0.364
0.383
0.02
0.013
0.027
0.006
0.005
-------
320
HID 16
• >0.2//
¦ > 1/U
A < \fi, > 0.2fi
90
80
70

50-
30-
20
10-
Hours
Figure 1. Kinetics of orthophosphate uptake by three par-
ticulate size classes at station A2, 7-5-78. The sample
was collected at 1 m below the surface where the
= 0.59 pmole-liter--*-.	wag usecj as the tracer.
Symbols and bars represent the mean and ranges of duplicate
determinations. Total uptake (>0.2 pm particle size): •;
uptake by the >1.0 jam size class: ¦; bacteriopLankton up-
take (<1.0 um, >0.2 pro particle size): a.
-------
HID 17
321
70-
60-
50
40-
«•
o-
10-
0-»	^ ^
Hours
Figure 2. Kinetics of orthophosphate uptake by 3 particu-
late size classes at station AO, 7-5-78. The sample was
collected at 1 m below the surface; [P04~2] = 0.78 umole"
liter~l. H333PO4 was used as the tracer. Symbols and
bars represent the mean and ranges of duplicate determi-
nations. Total uptake {>0.2 urn particle size): •; uptake
by the >1.0 yun size class: ¦; bacterioplankton uptake
(<1.0 pm, >0.2 nm particle size): & .
-------
322
HID 18
50-
40-
• : 1 meter
O : 3 meter
¦ : 6 meter
~ : 9 meter
i : lO meter
50-
40-
Hours
Figure 3. Kinetics of orthophosphate uptake by the total
microbial populations at 5 depths at station A2, 8-16-78.
H-j32p04 was used as	tracer. Symbols and phosphate
concentrations at each depth: 1 m - 1.12 pmole PO4-2•liter-1
•; 3m- 1.22 pmole PO4-2•liter-1, o; 6 m - 0.97 pmole PO*-2
•liter-1, ¦; 9 m - 0.88 ;amole PO4-2 • li ter-1, ~; 10 m - 0.96
jomole PO4-2•liter-1, ~.
-------
HID 19
323
Phosphate Concentration ;umole PO^ • liter	•
1.30
0.90
l.OO
080
l.tO
0.70
4-
I
X
+>
a.
e
o
9-
10-
8.0
2.0
Uptake Rate
6-0
10-0
liter
18 19
1J 16
17
9
10 II 12 13
14
Bacterial Standing Stock (* '0® « L ')
Figure 4. Depth profile of bacterial numbers (a), phosphate
concentrations (•) and uptake rates (¦) at station A2,
8-16-78. H, PO^ was the tracer. Rates are slopes of the
kinetics of orthophosphate uptake by the total microbial
populations at 5 depths. Bacterial numbers are from di-
rect counts by epifluorescent microscopy.
-------
324
HID 20
320-
Hours
Figure 5. Kinetics of orthophosphate uptake as retained on
an 0.2 pm pore size filter at station A2 and AO, 9-6-78.
Samples were collected from 1 m below the surface.
was used as the tracer. Phosphate concentrations were 0.78
jimole PO^-^-liter-! at AO (•) and 1.25 pmole PO*-2•liter~l
at A2 (¦) .
-------
HID 21
325
Figur* 6.
Figur* 7.
e
>
<3
o
0
&
o
X
fi.
«
0
1
0
X
«
JC
&
3
Figur« I
I	1
I 2
IOO-i Figure 9.
Figure 10
Filter Pora Sii» (,urn)
Figures 6-10. Percentage uptake retained on 5 pun, 1 urn and
0.6 Jim filters relative to that retained on an 0.2 v™ fil-
ter after a 24 h incubation period.	3PC>4 was the tra-
cer in the 7-5-78 experiments (•) , was used in the
8-16-78 experiment at station A2 (a) and xn the 9-6-78 ex-
periments (¦). All samples are collected from a depth of
1 m "below the surface.
-------
326
HID 22
B6'
WILMINGTON
LONG BEACH
cs«
,C3«
A11
D1»
B3*
C2
A7.
SAN
PEDRO
B10*
Bit"
A16*
B9»i
AM.
AS
A17»
V°
A13»
B1«
A9-
yol
AO
Harbors Environmental Projects
University of Southern California
Figure 11. Stations for Ortnophosphate Study
-------
HIE
327
COMMUNITY METABOLISM OF TOTAL ADENYLATES BY THE
MICROHETEROTROPHS OF THE LOS ANGELES HARBOR AND
SOUTHERN CALIFORNIA COASTAL WATERS
INTRODUCTION
The radiotracer method for studying uptake kinetics by
natural populations of aquatic microheterotrophs (Wright and
Hobbie, 1966; Hobbie and Crawford, 1969) has yielded consider-
able information on the turnover times and relative uptake
rates of various dissolved organic substrates. When combined
with a direct measurement of the natural substrate concentra-
tion, this kinetic approach can establish more precise values
of uptake velocity. If the standing stock of microhetero-
trophs is also measured, the specific activity of a given pop-
ulation for uptake of a particular compound can be calculated.
Wright (1978) has suggested three specific activity indices:
vmax'-,as 10~12ug*h~l-cell"*l; turnover rate (Tr) , as 10~6'h~l*
cell-1 liters; and direct uptake rate, Vn, as lO'^^^g.^-l.
cell-l. Data reported as a specific activity allow one to make
direct comparisons among aquatic heterotrophic systems, whether
they vary in space or time. The purpose of this study was to
compare seasonal variations in uptake activity by the bacterio-
plankton of the Los Angeles Harbor with the activity in the
contiguous coastal waters.
The adenylate system is well suited as a study of microbial
activity, since the adenylates occur as universal components of
all living cells, both in stable macromolecules like DNA and
RNA and as major chemical species involved in cellular energy
metabolism. Furthermore, adenylates can be measured in nanomo-
lar concentrations in the ocean, by means of the sensitive lu-
ciferin-luciferase assay (Holm-Hansen and Booth, 1966) ; Hodson,
ei al. , 1976). Previously, dissolved ATP has been shown to oc-
cur in the ocean at concentrations ranging from 0.1 to 0.6pg-li-
ter--'-, where it is rapidly utilized by marine bacteria (Azam and
Hodson, 1977).
^H-AMP is employed as a tracer of TA metabolism by micro-
heterotrophs in the present study. Its specific activity was
calculated on the basis of the total dissolved adenylate concen-
tration (TA = ATP + ADP + AMP), since it is probable that all
adenylates share the same transport system (Martin and Demain,
1977). However, this assumption will soon be tested under con-
trolled conditions with bacterial isolates from the harbor.
This report attempts to correlate direct counts of bacterio-
plankton standing stocks and measurements of natural adenylate
concentrations with the kinetics of TA uptake during three sea-
sons of the year (summer, fall and winter, 1978; the annual
-------
328
HIE 2
cycle will be completed by spring sampling in March, 19 79) at
a sampling station (A2) inside the Los Angeles Harbor and one
(AO) outside the breakwater in coastal waters. Previous stud-
ies for the uotake of other dissolved organic substrates {e.g.,
^4C-glycine, -l-^c-amino acid mixture, 14C-glucose) have shown
a 5- to 20-fold difference in uptake, on a relative basis, be-
tween these stations; however, this is the first time that up-
take of a single compound has been followed continuously over
an annual cycle in conjunction with measurements of its in situ
concentration.
METHODS
Seawater samples were collected by Niskin sterile-bag
devices from 1 meter below the surface and maintained within
2°C of the in situ temperature until return to the laboratory,
where they were filtered through a 20 3pm Nitex net to remove
larger plankton. All other filtrations employed discrete pore-
size membrane filters (Nuclepore, 47mm diameter) which are
hereafter designated simply as x pm filters, where x is 0.2,
0.6, 1 or 5pm and indicates the diameter of the pores. The
samples were stored at 18°C and assayed within 2 hours of re-
turn to the laboratory.
Extraction and measurement of ATP and TA were according
to the procedure of Holm-Hansen and Booth (1966) as modified
by Azam and Hodson (1977) and Karl and Holm-Hansen (1978). The
dissolved TA concentration is defined as that passing an 0.2pm
filter. The particulate TA of size fractions 0.2pm to 1.0pm
and 0.2pm to 203pm were estimated by measuring the lyim and 203pm
filterable TA, respectively, then subtracting the average TA of
the dissolved fraction.
Determination of bacterial and phytoplankton standing
stocks were by the acridine orange direct counting technique
(AODC) according to Daley and Hobbie (1978) and chlorophyll a
biomass estimates by the fluorometric technique of Strickland
and Parsons (1972).
Two different types of uptake experiments were performed.
For simple kinetics, uptake of label over time was studied. For
Michaelis-Menten (M-M) kinetics, varying concentrations of cold
AMP were added to different assay flasks and the velocity of up-
take at each substrate concentration was determined from a sin-
gle point incubation. Zero hour (to) and acid-killed controls
did not yield significantly different values and were generally
less than 10% of the uptake of label after 2.5 hours incubation
for any substrate concentration. For convenience, tQ blanks
were prepared for the simple kinetic experiments, while acid-
killed controls were employed in the M-M kinetics experiments.
Subtraction of these blanks eliminates nonbiological phenomena
such as adsorption and background radiation from inclusion in
the uptake data.
-------
HIE 3
329
the uptake data.
For the measurement of simple uptake kinetics, 50 pCi^H-AMP
(15 Ci-mmole"!) were added to 500ml seawater sample and incuba-
ted with stirring at 18°C. Duplicate 10ml samples were removed
at half-hour or hourly intervals for up to 7 hours and passed
through an 0.2pm filter. The filter was rinsed twice with 5ml
of 0". 2pm prefiltered sea water (SW) , dried for one hour under
an IR lamp, and placed in 10ml of a toluene-based fluor for
counting in an LS-100 liquid scintillation system.
In the first experiment (June 7, 1978) incorporation of la-
bel into macromolecules was measured by following the filtration
of duplicate 10ml samples with two 5ml rinses of OC 0.5N PCA
prior to the SW rinses. In the second experiment (August 2,
1978) duplicate 10ml samples were passed through 1.0pm filters
so that kinetics of uptake by the >1.0 pm size class, and by
subtraction, by the >0.2pm, <1.0pm size class of microhetero-
trophs, could be followed in addition to total uptake.
For the M-M kinetic experiments) 10ml aliquots of sample
were dispensed in duplicate into sterile serum bottles (100ml
capacity) to which a 1ml volume of substrate containing 1 pCi
3h-AMP and varying concentrations of cold AMP was added, for a
final concentration of 6 to 100 nmole TA-liter"!. In the last
experiment (December 6, 19 78) the addition of 3H-AMP was re-
duced to 0.1 pCi to yield a low concentration of 0.6 nmole TA-
liter~l. After a 2.5 hour incubation (2 hours at A2 and 5 hours
at AO on December 6, 1978), the samples were taken through the
filtration procedure described above. Acid-killed controls, to
which the addition of 0.02ml of a 7% PCA solution immediately
preceded the addition of label, were filtered after the same in-
cubation period. In the first two experiments, controls were
prepared only for the lowest and highest substrate concentra-
tions; thereafter they were prepared for all concentrations.
For the size-fractionation studies, 100ml of water sample
were incubated with 10 pCi of ^h_amp at 18°C for endpoint deter-
mination of the uptake rate by four size classes of microhetero-
trophs. Acid-killed controls were prepared for each sample.
After 24 hours (4h at AO and 2h at all other stations on Decem-
ber 6, 1978) duplicate samples were removed for filtration
through 5.0, 1.0, 0.6 and 0.2 pm filters, and treated as de-
scribed above.
Data Calculations
Conversion of counts per minute (cpm) to disintegrations
per minute (dpm) was by means of a quench curve relating exter-
nal standard ratio to counting efficiency. Specific activity of
the label was calculated, based on the sum of the added 3H-AMP
and the natural TA concentration in the seawater sample. One
ml aliquots of sample plus label were counted directly in Aqua-
sol to determine total dpm (3h-AMP) •ml"-'- of sample.
-------
330
iiie 4
For the simple kinetic studies, uptake values, as nmole
TA'liter-!, were plotted against incubation time. The poten-
tial uptake rate (Vp), at the elevated (TA + 3h-amP) adenylate
concentration, is the slope of the line determined by linear
regression analysis, and is given as nmole TA*liter"!'h~l.
The specific potential uptake rate (VpS) is determined as the
uptake rate per 10^ bacterial cells (by AODC). The turnover
time (T^) for this elevated TA concentration is determined as
the quotient of the rate (V) divided into the sum of the nat-
ural TA + %-AMP concentrations.
For analysis of the M-M kinetic data, the uptake rate at
each concentration of substrate was plotted against substrate
concentration to determine whether saturation kinetics occurred.
A Woolf transformation of this data, to an S/v vs S plot, yields
a straight line with the equation
S/v S/Vmax + Vvmax
where
= a concentration constant similar to the Michaelis
constant Km = the negative abscissa intercept
Vmax = a velocity constant observed when a limiting step
is saturated with substrate = the inverse of
the slope
S = substrate concentration
Vv
= the turnover time for substrate at each concentration
The turnover time (Tt) at the natural TA concentration, is
the time required for the microheterotrophs to remove all
adenylates from solution (assuming no further input) and is de-
termined as the ordinate intercept of the Woolf plot. Turn-
over rate (Trn) is the inverse of T^ and is reported here on a
per cell basis, as suggested by Wright (1978), as 10~6-h-1*
cell"1•liter~l. The uptake rate at the natural concentration
of substrate, Vn, was calculated by substituting the natural
TA concentration for S in the equation above, and solving for V.
Specific uptake rate at the natural concentration (Vns) was cal-
culated as nmole TA*10^ cells"!•h~^ (cell number from AODC).
Vmax was also calculated on a per cell basis.
For analysis of the endpoint size fractionation data, up-
take rates were calculated for each size class as nmole TA-li-
ter""l-h-1. The percent uptake passing a given filter porosity
was determined, assuming a 100% retention by the 0.2pm filter.
RESULTS
The uptake of dissolved %-AMP was studied in June, August
and December, 1978 at station AO and October as well at station
-------
HIE 5
331
A2 in the Los Angeles Harbor. The dissolved natural TA con-
centration ranged from 1.19 to 1.74 nraole TA*liter"! at sta-
tion AO and from 1.5 3 to 4.96 nmole TA*liter-1 at A2 for these
months, while the particulate TA concentration (>0.2pm, <203 pra)
ranged from 1.89 to 5.75 nmole TA*liter~l and from 3.47 to 19.8
nmole TA-liter~l, respectively, at AO and A2 (Table 1).
The kinetics of %-AMP uptake was linear over 7 hours at
both stations, as shown in Figures 1-3. At station A2, poten-
tial uptake velocities per liter of water were high in June and
August compared with uptake in December, at 1.08, 0.8 7 and 0.2
nmole TA»liter-!, respectively. However, on a per cell basis
these values become 0.57, 0.49 and 0.43 nmole TA-10 cell"!*h~!,
with a variance less than 25% among them. The bacterial stand-
ing stocks are shown in Table 2. Specific potential uptake
rates at AO are less than half those at A2 on the same sampling
date. This difference is greatest in August, when the potential
uptake rate at A2 is 10 times as high as at AO (Table 3, 12th
column).
Turnover times likewise show wider disparities among sam-
pling dates at a given station than do turnover rates, which
are reported on a per cell basis. The longest turnover time
occurred at AO in August, at 258 h, while the shortest was at A2
in June at 10 h. These differences become much smaller when ex-
pressed as turnover rates of 6.1-10~6-h-!-cell-l-liters and
54.3•10~®•h~1•cell~l-liters, respectively. The dissolved TA
concentration at AO was, on the average, 60% lower than at A2
and varied less than 20% from its average value of 1.45 nmole
TA-liter-!. The December adenylate concentration at A2 was
only 40% of the mean value of 3.69 nmole TA-liter-! over these
three months.
The rate of incorporation of %-AMP is not significantly
different from that of uptake; at neither station do these rates
differ by more than 0.003 nmole TA•liter~l•h-!. After five
hours of incubation, over 96% of the assimilated label is appar-
ently incorporated into macromolecules; soluble pools must be
quite small. Azam and Hodson (1977) reported a 98% assimilation
of l^c-ATP Uptake in the Saanich Inlet, British Columbia, indi-
cating that little is respired. Assuming that respiration is
also negligible in this system, it is concluded that over 90% of
the adenylates transported across the cell membrane result in
macromolecular incorporation.
In August (Figure 2) we examined the uptake kinetics of
H-AMP into two size fractions. A surprisingly high (33-49%)
percentage of total uptake was associated with the size class
>1.0pm, presumably containing phytoplankton, microzooplankton
and bacteria attached to particles. This contrasts with the
predominance (80-90%) of uptake by the size class >0.2pm,
<1.0^m = bacterial, for !4c-glucose, a l^c-amino acid mixture
from algal hydrolysate, and %-thymidine, as found in an ear-
lier study (Sullivan, et al. , 1978) and for dissolved glucose,
-------
332
HIE 6
serine, acetate and AMP in sea water (Azam and Hodson, 1977) .
The results of the single point size fractionation studies
(Figures 4 and 5} show that the microheterotrophs in the size
fraction <1.0pm generally accounted for more than 75% of the
%-AMP uptake. However, in June and August, months immediately
preceding phytoplankton blooms, this fraction contained 40 to
50% of the %-AMP taken up. By contrast, this fraction only
contains between 10 and 30% of the particulate adenylates
(Table 1).
Figures 4-7 show the results of the multiconcentration
experiments for ^H-AMP uptake at stations A2 and AO. Addition
of cold AMP plus 3H-AMP to the samples resulted in concentra-
tions ranging from 0.4 and 1.6 times that of the natural adenyl-
ate concentration to concentrations two orders of magnitude
greater. The shape of the S vs V plot varied from month to
month at station A2 (Figures 4 and 5a and b). In June and Octo-
ber an initial saturation seems to be followed by a possible
second increase in uptake at higher substrate concentrations.
This was not the case in August, when at very high concentra-
tions the %-AMP uptake rate decreased with increasing S, or
in December, where the plot is more linear than hyperbolic.
Woolf transformations of these data into S vs S/v plots
are shown in Figures 6 and 7. All plots were linear, with
correlation coefficients >0.88 at the cx = 0.01 level. In the
Woolf transformation of the August experiment (Figure 6b) the
line was calculated for all concentrations but the two highest;
as inclusion of the latter points would have resulted in a neg-
ative turnover time. These points (circled in Figure 6b) cor-
respond to the rate decrease in Figure 5a and are considered
to have arisen from experimental error.
It is constructive to note the behavior of the S vs S
plots as they near the ordinate axis. At station A2 in June,
August and October (Figure 6) the S/y values at the lowest con-
centrations (single point turnover times) are greater than the
value of the ordinate intercept (multiconcentration turnover
time). This conforms to the theoretically ideal situation, and
is what can be expected in eutrophic waters (Gocke, 1977), such
as occur in the Los Angeles Harbor. However, in the December
experiments at both stations AO and A2, the S/y values curve
towards the abscissa axis at lower concentrations. Apparently,
this phenomenon is often associated with more oligotrophic
waters, where uptake by a heterogeneous population results in
computation of longer turnover times by the multiconcentration
method (Williams, 1973). December is the winter season in the
harbor and both the bacterioplankton population and in situ
adenylate concentrations are <25% of their October values at
station A2. This is consistent with multivariate benthic and
plankton analyses, where outside station Al and outer harbor
stations cluster together in December, but are separate the rest
-------
11 IE 7
333
of the year. Station AO is just outside the harbor breakwater
and even in summer months is characterized by lower microbial
standing stocks and nutrient levels than occur inside the har-
bor (Tables 2 and 3}.
The uptake constants Kfc and Vmax are summarized for each
experiment in Table 3. Vmax values ranged from 0.3 nmole TA-
lO^ cells~!-h~! at station A2 in October to almost 2.0 nmole
TA-10^ cells-l-h~! at station A2 in June and December. Kt val-
ues ranged from a high of 105 nmole TA-liter~l at station A2 in
December to a low of 11.9 nmole TA-liter-! at A2 in October.
Turnover times determined both by simple and M-M kinetics
are also compared in Table 3. The former method yielded values
30 to 65% lower than did the latter, except in August. Part
of the explanation for this finding is the difference in uptake
rate over seven hours as opposed to the first two hours of incu-
bation? both have correlation coefficients of >0.90, but the
initial uptake rate is only about half the rate over seven hours.
When turnover time is calculated on this lower rate of uptake,
the result is much closer to that calculated from M-M kinetics,
where the incubation period was 2-5 hours.
The rate of uptake of dissolved ^h-amp at the natural TA
concentration (^n) as calculated from M-M kinetics, is shown in
the last two columns of Table 3. Unlike the potential uptake
rate (Vp), which showed little seasonal variation on a per cell
basis, the specific Vn rates vary tenfold, from 0,29 to 0.025
nmole TA*10^ cell~!-h~! in August and December, respectively.
Bacterioplankton and phytoplankton standing stock (biomass)
values are shown in Table 2. Note that the bacterial biomass
at station A2 is, on the average, almost three times that at
station AO. Phytoplankton standing stocks remain fairly con-
stant except in July, when a bloom occurred at all stations, and
in September, when a second bloom occurred in patches in the
harbor. The bacterioplankton increase, apparently in response
to these blooms, peaked in July and October. Total particulate
adenylate concentrations (Table 1) at stations AO and A2 peaked
around June-July and in September. Approximately 80% of the to-
tal particulate adenylate is found in the size fraction >1.0pm
throughout the period sampled.
DISCUSSION
Microheterotrophic activity in the Los Angeles Harbor and
coastal waters is high, with Vmax values ranging from 0.35 to
3.7 nmole TA-liter"!»h"! (0.13 to 1.3 pgC*liter~l•n~!). The
magnitude of these Vmax values is typical of highly eutrophic
waters, comparable to those found for glucose and leucine, 1.86
and 0.34 ugC•liter"!•h~!, respectively, in the Kiel Fjord
(Gocke, 1977) and higher than those measured in Lake Erken for
glucose, where Vmax values range from 0.009 to 0.072 pgc•liter-!
•h~! (Wright and Hobbie, 1966). The dissolved adenylate
-------
334
HIE 8
concentration is also high compared to other oceanic measure-
ments, e.g. dissolved ATP ranged from 0.22 to 2.90 nmole* liter-1
(118 to 1557 ng ATP-liter-1) at stations AO and A2 between April
and December, 19 78. This is comparable to the average dissolved
ATP concentration of 466 ng* liter"*1 in the eutrophic waters of
Saanich Inlet, British Columbia and of 218 ng'liter-1 off the
SIO pier at San Diego (Azam and Hodson, 1977) .
Since a minimum of 90% of adenylate uptake apparently re-
sults in incorporation into acid insoluble material (presum-
ably macromolecules), the uptake kinetics for this substrate
reflect on both the activity and growth potential of a popula-
tion. A comparison of the potential (Vp) and actual (Vn) uptake
rates at A2 (Table 3) reveals that, on a per liter basis, both
show substantial decreases in adenylate uptake from summer to
winter. Vp ranges five-fold, from 1.08 to 0.2 nmole TA-liter-1
•h"1 while Vn ranges forty-fold, from 0.51 to 0.013 nmole TA-
liter-1•h-1. However, when the VpS and Vns are considered, a
different result emerges. Whereas Vps differs little between
June and December, at 0.5 7 and 0.4 3 nmole TA-109 cell-1«h-1,
respectively, Vn= still varies tenfold, from 0.29 to 0.025 nmole
TA*10® cell-i-h-*.
These data indicate that, while potential uptake activity
changes primarily as a function of the microheterotrophic popu-
lation size, uptake rate at the in situ substrate concentration
varies in a more complex way annually. The tenfold difference
still seen between summer and winter Vns rates may be due in
part to subtle differences in the dissolved TA concentrations on
which these rates are based. Whereas the natural adenylate con-
centration at A2 decreased 3-fold, from 4.96 to 1.53 nmole TA-
liter-1 in August and December, respectively, the elevated con-
centrations for which Vp values are measured vary less than 1.5-
fold over this period.
When similar comparisons are made between the Vp rates at
AO and A2, it can be seen that the VpS at AO is 2- to 10-fold
lower than at A2 on a given sampling date; the widest disparity
in Vpg values occurs in August. Unfortunately, M-M-type kinetic
experiments were unsuccessful at AO in June and August; there-
fore only in December can Vn comparisons be made between the
stations. The Vns at AO was 0.014 nmole TA-1Q9 cell-1-h-1, a
little over one-half that at A2 (0.025 nmole TA-liter-109 cell-1
*h-1). This indicates that the bacteria outside the breakwater
are both potentially and actually less active in the uptake of
3h-amp than are those inside the harbor, even when the bacterio-
plankton densities are very similar at the two stations, as they
are in December (Table 2). This observation could be due to a
number of causes, such as different species compositions, differ-
ent transport capabilities, inducible transport systems, or a
higher percentage of dormant cells outside the breakwater.
The Vmax would be expected to vary seasonally in agreement
-------
HIE 9
335
with the VD rates. Specific Vmax values vary about 2-fold at
A2, which corresponds to the low specific Vp variation at this
station. The December Vmax value at A2 is about four times
that at AO in this month; the difference in their Vp values is
3-fold.
The turnover rate per cell is another measure of activity,
one that is strongly dependent upon the rate of substrate in-
put, of which we have no measure in this study. Although a
linear rate of uptake was assumed from a steadily diminishing
pool of the dissolved %-AMP in this study, a rapid cycling of
nutrients might actually have been missed in the closed environ-
ment of our flask. However, the brevity of the incubation peri-
ods compared with the uptake rates measured indicates this is
unlikely.
In the natural environment even less is known about the
rates of processes, such as grazing, excretion and cell lysis,
which lead to an input of TA into the dissolved fraction from
particulate matter. If a steady state is assumed for the par-
ticulate and dissolved adenylate concentrations at A2 from av-
erage values measured in this study, a first approximation can
be made of the rates between pools in the adenylate system, as
diagrammed below:
PARTICULATE
ADENYLATE
nmole TA* liter""'"
1.4
size >0.2um,
<1.Opm
8.8
size >1.0pm,
<20 3pm
Rates Kj_, K-^g, K_ip are in units nmole TA*liter-1*h-1.
Kj_ = rate of input of adenylates into the dissolved pool
from the particulate fractions
K-lB = uptake rate of dissolved adenylates by the bacterio-
plankton (<1.0pm size fraction)
K_ip = uptake rate of dissolved adenylates by the >1.0pm
size fraction
DISSOLVED
ADENYLATE
nmole TA*liter-1
K.m " 0.19
Kx = 0.24
size <0.2pm
K-1P = 0-05
Assuming that a steady state exists for particulate and
dissolved adenylate concentrations, then Ki = K_^ = K_1B + K_lp.
-------
336
HIE 10
K_^ is assigned the average value from measurements in this
study of 0.24 nmole TA-liter-I-h-l. Since the size fractiona-
tion data indicated that 50 to 90% of heterotrophic uptake of
3h-AMP is associated with organisms passing a l.Ojim filter, a
value of 80% the K_j, or 0.19 nmole TA-liter~^-h~l is assigned
as k-1B' leavirig 0.05 nmole TA-liter~l-h~l as K_ip. Thus,
= K_^ = K_1b + K_xp =0.24 nmole TA • liter-1 • h~*. When this
rate is divided into the particulate adenylate concentration,
a turnover time of 43 hours is estimated for particulate adenyl-
ates in the Los Angeles Harbor. The flux of particulate adenyl-
ates into the dissolved adenylate pool is the sum of many pro-
cesses/ major ones being excretion, decomposition and grazing,
with cell lysis and leakage being less important.
Although information is too limited to subdivide the K]_
flux, it is clear that the bacterioplankton predominate in the
uptake of dissolved adenylates. From the kinetic experiment in
which adenylate uptake was fractionated, Vp values for the
>0.2pm, cl.Opm size class and >1.0pm size class were 0.46 and
0.42 nmole TA-liter~l-h~l, respectively. .If we assume that all
the uptake by the larger size class is algal, an all the uptake
by the smaller size class is bacterial, then specific Vp values
for this experiment are 0.0006 and 0.032 nmole TA-pgC-h 1, re-
spectively. Thus, the bacterioplankton uptake exceeds the phy-
toplankton uptake rate by 55 times, on a per biomass basis, al-
though the two are of equal magnitude on a per liter basis.
This comparatively high uptake by the larger size class may al-
ternatively be interpreted as being primarily due to bacteria
attached to particles. Particulate organic matter may have been
relatively high at the time of this sampling (August) following
the July bloom of Nitzschia seriata throughout the harbor.
Although no estimate can be made of the relative rates of
input into the dissolved TA pool from the two particulate size
classes, it can be noted that the standing stock of the larger
size class is 6 times that of the cl.Opm size class. This in-
dicates the formation of particulate adenylates by other means
than through uptake of dissolved adenylates. Of course, the
major processes of grazing, photosynthesis, and biosynthesis
have been omitted from the overly simplified budget.
A' comparison of the natural dissolved TA concentrations
(which range from 1.53 to 4.96 nmole TA-liter~l) with the K-j-
values derived from M-M kinetics (which range from 11.9 to 105
nmole TA-liter~l, Table 3) indicates that the microheterotrophs
in these waters are always substrate-limited for TA; their up-
take rate at the natural concentration being 1 to 30% of the po-
tential Vmax rate at saturating levels of TA. These extremes
occur in December and August, respectively, months in which the
disparities between the natural TA concentrations and the Kt
values are greatest and least, respectively. The value at
station AO in December is only 40% that at A2, which indicates
that the bacteria outside the harbor may have a higher affinity
for their substrate in partial compensation for their lower
activity.
-------
I HE 11
337
SUiMMARY
The following summarizes the results of a study on the
in situ concentrations and uptake rates for dissolved total
adenylates (TA = ATP + ADP + AMP) by the microheterotrophs of
the Los Angeles Harbor and coastal waters.
1.	The natural TA concentration at station A2 inside the
harbor ranged from 1.5 to 5.4 nmole TA-liter~l, whereas
the Kt value ranged from 11.9 (August) to 105.4 (Decem-
-ber) nmole TA-liter-!. At station AO outside the break-
water, dissolved TA concentration ranged annually from
1.2 to 2.2 nmole TA•liter~1; the K. value for December
was 41.8 nmole TA-liter~l. These data indicate that the
transport system of microheterotrophs for adenylate up-
take is always undersaturated in these waters.
2.	Vmax and Vp values are similar in measuring the poten-
tial rates of uptake a population shows for substrate
concentrations above the in situ level. At station A2
these rates vary seasonally by 4- and 5-fold, respec-
tively, when expressed as nmole TA-liter -h~l. When ex-
pressed on a per cell basis, however, the annual varia-
tion is 2-fold or less. These data indicate that uptake
potential is directly correlated with the number of bac-
teria present.
3.	Vp, the potential uptake rate, is 2 to 10 times greater
than the uptake rate at the natural substrate concentra-
tion, Vn, which is derived from M-M kinetics. Specific
Vn varies 10-fold between August and December at A2,
which indicates that the actual uptake activity of the
microheterotrophs at the natural substrate concentration
varies on a seasonal basis. It seems more likely that
this variation would be due to subtle differences in
in situ TA concentration than to temperature effects,
since all experiments were carried out at 18°C, and
since Vp values would be equally expected to vary with
differences in temperature.
4.	The specific Vp at AO is 2- to 10-fold lower than at A2
on a given sampling date and the specific Vn for Decem-
ber at AO is only half that at A2. This indicates that
the bacteria outside the harbor breakwater are less ac-
tive in the uptake of adenylates than are those inside
the harbor, which could be explained by differences in
species composition, transport capabilities for the
adenylates, or metabolic status of the populations.
5.	A simple budget was made for the TA flux between the
-------
338
HIE 12
dissolved and particulate pools in the harbor. Assuming
a steady state concentration of 10.2 nmole TA-liter~l
particulate adenylates and 3.5 nmole TA*liter-! dis-
solved adenylates, the rate of input of particulate TA
into the dissolved fraction is equal to the rate of up-
take of dissolved adenylates into the particulate frac-
tion, 0.24 nmole TA*liter"Such complex processes
as grazing, leakage, excretion, and cell lysis are
lumped together here as potential sources of the dis-
solved TA.
6. Size fractionation studies indicate that approximately
20% of the particulate adenylate concentration is found
in the >0.2, cl.Opm size class, which generally account-
ed for 75% of the ^h-AMP uptake. This shows that bac-
terioplankton are predominant in the uptake of dissolved
TA; however, in June and August, months which immediate-
ly preceded phytoplankton blooms, only 40 to 60% of the
uptake passed a l.Ojim filter, which suggests that phyto-
plankton may also be active in uptake at this time.
LITERATURE CITED: See Section VI.
-------
HIE 13
339
Table 1. Dissolved arid particulate TA concentrations in the Los Angeles
Sampling
date
Harbor and coa
Size fraction
stal waters
Station
AO I A2 1 A7 1 A12 1 B9
Total adenylates (nmole«liter )
6-7-78
<0.2y
0.2y-l.Oy
0.2y-203y
1.0y-203y
1.74
0.33+0.7
5.75±0.6
5.4
3.97±1.21
1.23±0.34
19.8±3.3
18.6
4.29±0.16
25.6+5.3
4.14+0.41
0.49+0.4
3.48±0.11
3.0
3.09+0.16
2.04±0.30
14.8±4.0
12.8
8-2-78
<0.2y
0.2y-l.Oy
0.2y-203y
1.0y-203y
1.41
0.75±0.03
2.45±1.48
1.7
4.96±0.48
1.89+0.51
6.10±0.10
4.2
6.24+1.05
7.89
24.2+1.0
16.3
2.27±0.12
2.81±1.03
8.21±1.19
5.9
2.35±0.13
3.78±0.30
12-6-78
<0.2y
0.2y-1.0y
0.2y-203y
1.0y-203y
1.19±0.10
0.17±0.17
1.89±0.44
1.7
1.53±0.45
1.08
3.47±1.47
2.39
2.35±0.52
0.94±0.50
3.00±0.20
2.1
1.70±1.23
0.64±0.34
8.12±2.68
7.5
1.76±0.17
3.71+0.35
4.09±0.06
0.4
* The particulate adenylate of size fractions 0.2p to l.Op and
0.2vi to 203p were estimated by measuring l.Op and 203p filter-
able adenylate, respectively, then subtracting the average
adenylate of the 0.2p (soluble) fraction.
-------
Table 2. Bacterioplankton and phytoplankton standing stocks at stations A2 and AO, 1978
Stations
Sampling date
9 B' -1
10 cells-liter
'
AO
ygC-liter
" 1
i
p3 -i
pgC'liter
9 ® -1
10 cells'liter
A2
62 -1
jjgO liter
p3 -1
ygC-liter
June 7
0.75 + 0.08
6.0
87.8
1.88 ± 0.14
15
309
August 2
0.64 ± 0.7
5.0
70.4
1.79 ± 1.5
14
727
October 194
-
-
_
3.634
294
340
December 6
0.7 ± 0.13
5.5
377
0.47 ± 0.14
3.8
172.5
Method of calculation of data:
^Bacterioplankton number of AODC ^one SD
Bacterioplankton biomass (see part IIIA of this report).
3
Phytoplankton biomass (see part IIIA, this report).
4
Average of October 4 and November 6 values.
-------
Table 3. Summary of the uptake velocities, turnover rates arid times, arid kinetic constants KT and V
at stations A2 and AO, 1978, both from simple and M-M kinetics.	max
Sampling
date
1978
Station
Sn
nmol
TA-L"1
V
max
nmol
TA-L"1
V
max
nmol
TA -109
cell~l
kt
nmol
TA-L"1
V
h
T b
r
10-6-h-J-
cell
•L-l
Tt°
h
lo-^h:1 1
cell-L
VPS
nmol
TA*L-1-h-1
vps
nmol*TA
celT-h"1
v f
n
nmol-TA
L-V1
vns
nmol TA-109
cell~*-h-1
6-7
AO
1.74
-

_
-
-
42.5
31.4
0.20
0.26
-
-
6-7
A2
3.97
3.7
1.97
65.4
17.7
30.0
9.8
-
-
-
-
-
8-2
AO
1.41
-
-
-
-
-
258.4
6.1
0.03
0.05
-
-
8-2
A2
11.96
1.75
0.98
11.9
6.8
82.1
13.2
61.4
0.87
0.49
0.51
0.29
10-19
A2
5.709
1.06
0.29
27.8
26.1
10.5
-
-
-
-
0.18
0.05
12-6
AO
1.19
0.35
0.50
41.8
120.8
11.8
82.0
17.4
0.10
0.13
0.009
10.014
12-6
A2
1.53
0.93
1.98
105.4
112.8
18.9
39.8
53.4
0.20
0.43
0.013
0.025
Method of calculation:
a.	ordinate intercept on Woolf plot
b.	turnover rate; inverse of Tj. in a
c.	slope of simple kinetic curve divided into TA concentration
d.	turnover rate; inverse of Tt in a
e.	potential uptake rate; derived from simple kinetics
f.	uptake rate at material TA concentration; derived from M-M kinetics
g.	mean of dissolved TA measured 10-4-78 and 11-6-78
- no data
All other determinations are as described in methods.
-------
342
HIE 16
7.0-]
6.0
S.O
>»
3.0-
2.0
1.0
0-i
O.S
Hours
Figure i. Uptake (•) and incorporation (a) of 3h-AMP at
STATION A2, June 7, 1978.
-------
HIE 17	343
7.0
6.0
5.0
4.0
x
30
3.0
1.0
0
3
4
5
6
1
2
7
Hours
Figure 2. uptake of 3h-AMP at station A2, August 2, 1978
INTO 3 SIZE CLASSES: TOTAL(•, >0.2 pM), >1.0 MM
<~) AND >0.2 liM, <1.0 MM (A).
-------
344
IIIE 18
2.0-
7J0-
t «H
1.0-
2.0-
1.0
3a
Aq June 7, 1978
.**-5he
Aq August 2,1978
y = 0.031 x - O.Ol corr » Q.974 T» = 258h
Ag December 6,1978
y »o.095*
- 0,983 Tf8«h
A2 December 6,1978
Hours
.Figure 3a. Uptake (•) and incorporation (a) of 3h-AMP at
STATION AO, June 7, 1978.
3b. Uptake of 3H-AMP at station a2, August 2, 1978
INTO 3 SIZE CLASSES! TOTAL (•, >0.2 MM),
>1.0 yM (~) AND >0.2 pM, <1.0 UM (A).
3C. UPTAKE OF 3H~AMP AT STATION AO, DECEMBER 6, 197
3d. Uptake of 3H-AMP at station A2, December 6, 197
-------
HIE 19
345
6-7-78
nmote adonylates . liter
a
D
1.0-
0.8-
0.6
0.4-
0.2-
4b
A2 10-19-78
V
/
° •
0 20 60 lOO 140
nmole adenylate*«u'fer - [Sj
Figure 4a. m-M kinetics for uptake of ^H-AMP at station A2,
June 7, 19 78.
4b. M-M Kinetics for uptake of 3H-AMP at station A2,
PCTOBER 19, 1978.
-------
346
HIE 20
2.O-1
J.O-
s
X
c
«
¦O
0
•
o
E
e
5o
A2 8-2-78
i"
Uo
260
O 20 40 60 SO too
ISO
240 280
20H
>•
Jt
e
«-
a.
s
1.0
O.S-
0.6-
o.«-
0.2-
5b
Aj 12-6-78
• •
5«
A0 12-6-78

~1—1—1—1—1—
O 20 40 60 SO 100
—I—> 		f	1	,	1	1—
160 0 20 40 60 SO IOO
—I—
160
nmole adenylates* liter*' 00
Figure sa.
5b.
5C.
M-M KINETICS
August 2, 19
M-M Kinetics
December
M-M Kinetics
DECEMBER 6,
FOR UPTAKE
78 .
FOR UPTAKE
1978.
FOR UPTAKE
1978.
OF 3H-AMP AT
OF 3H-AMP AT
OF 3H-AMP AT
STATION A2,
STATION A2,
STATION AO,
-------
XXIE 21
347
6a
6-7-78
100-

6b
200-
100-
6c
A- 10-19-78
.v
lOO-
u
a
o
X
80 lio 160 200
S nmole adenylates • liter '
40
280
Figure 6a. Woolf transformation (WTD) of M-M kinetics for
uptake of 3H-AMP at station a21 June 7, 1978.
6b. Woolf transformation (WTD) of M-M kinetics for
uptake of 3H-AMP at station a21 August 2, 1978.
6c. Woolf transformation (WTD).of M-M kinetics for
uptake of 3h-AMP at station A2, October 19, 1978.
-------
348
HIE 22
600-
500—
400-
X 300-
200-
100-
0-
40
80 120 160 200
nmole Adenylates • Iit®r [S]
280
240
Figure i. Woolf transformations of m-m kinetics for uptake
OF H-AMP AT STATIONS AO AND A2, DECEMBER 6, 1978.
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IV
349
INTERACTIONS OF PHYSICAL AND BIOLOGICAL PARAMETERS
INTRODUCTION
The need for integration and evaluation of large amounts
of data has increasingly required investigators to resort to
statistical analytical techniques- Biological systems are
generally more variable and less predictable than physical or
chemical systems; furthermore, the physico-chemical factors in
the environment strongly affect biological systems, controlling
such things as reproductive periods, food chain sequences and
distribution patterns. Attempts to identify and quantify the
interactions are still dependent upon the input of biological
expertise from a variety of fields- However, analytical computer
methods provide the means for integrating large amounts of data
for multiple parameters and for identifying which parameters have
exerted the most influence on the ecoysstem at a particular time.
.METHODS
Smith (1976) developed methods for ecological analysis and
the use of weighted discriminant techniques, some of which were
used in the first ecological study of the entire Los Angeles-
Long Beach Harbors in 1973-1974 (AHF, 1976).
In the following pages hierarchical classification was used
to study patterns of the biological data. Groups of biologically
similar sampling sites (stations) were defined and the groups
developed from the biological composition of the sites were then
compared with the patterns of measured environmental parameters.
From this, hypotheses concerning the relationships between the
biota and the environment were suggested.
Flexible Sorting (B=.25) Strategy (Lance and Williams, 1967)
and the Bray-Curtis Distance Index (Bray and Curtis, 19 57;
Clifford and Stephenson, 1975) were used to classify sampling
sites.
The relationships between the species and the station
groups defined by classification (dendrograms) were examined
in two-way coincidence tables (TOT) (Kikkawa., 1968; Clifford
and Stephenson, 1975). The numbers in the body of the table
were transformed and standardized, and converted to symbols of
species maxima as follows:
* >	. 7 5 to 1
+ >	.5 to .7 5
- >	.2 5 to .50
>	0 to .2 5
blank
0
-------
350
IV 2
To test for complex biotic-environmental relationships,
the groups were examined by weighted discriminant analysis
(Smith, 1973). Because of the technical nature of these
analyses, Smith's (1978) paper is appended herein as section
VIB.
Because taxonomic studies deal with only the most common
identifiable species under some circumstances, and identifica-
tion of rare and little known species under others, the data
analyzed in the following sections were restricted so that
comparisons between seasons and years could be made in a
uniform manner. Thus the zooplankton analyses were restricted
to copepod and cladoceran species (by far the most numerous
in species and populations) and the benthic analyses were
restricted to species of polychaete worms and molluscs.
Circulation in the outer Los Angeles Harbor is dominated
by a large gyre, which appears to rotate much of the time in
a clockwise fashion on the surface, and probably in a counter-
clockwise manner at depth (Robinson and Porath, 1974) . The
patterns tend to persist through tidal cycles, although they
have been observed to break up during shifts from the prevail-
ing southwest winds to high so-called Santa Ana Winds from the
east. The gyres have been reproduced in the U.S. Army Engin-
eers physical model of Los Angeles-Long Beach Harbors, in
Vicksburg, Mississippi. Figures 1 and 2 illustrate representa-
tive conditions on incoming and outgoing tides, respectively
(from McAnally, 1975).
Circulation patterns and flushing rates govern the distri-
bution and assimilation of wastes and nutrients in the harbor.
They also affect the sorting and deposition of variously sized
sediment particles, which in turn affect the habitats of benthic
organisms. Circulation serves to distribute the planktonic
larvae (meroplankton or temporary plankton) of indigenous
organisms, and tidal exchange brings both larvae and adult
zooplankton into the harbor. The distribution patterns developed
in the station groupings for the following sections at times
show evidence for the influence of the main gyre and for a
transitory counterclockwise gyre in the western part of the
outer harbor.
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IVA
351
WEIGHTED DISCRIMINANT ANALYSIS OF ZOOPLANKTON
Zooplankton data, discussed in section IID, were examined
on a quarterly basis by discriminant analysis techniques for
the two-year period beginning in December 1976. At that time,
DAF-treated cannery wastes and primary-treated TITP sewage were
entering the harbor. The results of each period are discussed
in the following pages and illustrated for each seasonal quarter.
RESULTS
December, 19 76.
Stations for this period are well separated into an outer
harbor-outside harbor group, a shallower, nearshore group, and
the outfall area (A7) (Figures 3 and 4). The separations into
groups are made on the basis of species distributions and
numbers shown in the Two Way Table (TWT, Figure 5).
The weighted means of the physical and biological (phyto-
plankton) parameters used are presented in Table 1. Tabie 2
shows the coefficients of separate determination, in which
higher values provide indication of the important variables in
separating the groups. Generally coefficients of 10 or above
are considered important; the percent of information in each
axis is indicated on Table 2, and only coefficients on axes
with content of 1% or above are considered herein.
According to the coefficients, temperature, pH and chloro-
phyll a were the important factors of the parameters measured.
This does not discount the real possibility that, in some
instances, parameters not measured exert significant influence
and the station groupings will not be as clearcut as they were
in December 1976. Group 3 sites had the highest weighted mean
dissolved oxygen (DO), pH, primary productivity and assimilation
ratio, and the lowest chlorophyll a and salinity. The outfall
(Group 2) had the highest temperature and salinity, lowest DO
and pH, and lowest productivity and assimilation ratio. Group
1 sites were intermediate in almost all parameters and were also
intermediate in space. The important vectors and the station
groups are located on the axes in Figure 6. The data on
nutrients such as ammonia and nitrate were not included because
these are represented in the phytoplankton crop. The harbor
has not been considered nutrient limited in the past.
March 1977.
The explosion and Bunker C spill from the tanker Sansinena
occurred two weeks after the December field sampling. Analysis
-------
352
IVA 2
of the immediate area (A9, AlO) influenced by that event showed
that oil and grease levels in the water column were important
to the benthic and zooplankton populations in the western harbor
where 22 sampling stations were established in December 1976
following the blast (Soule and Oguri, 1978) . Oil and grease
measurements were not a part of the TITP study, but the March
pattern suggests a connection, probably tidally induced, for
Group 2 stations {Figures 7 and 85 . The TWT (Figure 9) shows
a considerable reduction in species or populations at Group 2
stations over the December TOT (Figure 3). On the other hand,
station A7 showed an increase, allying it with All as Group 3.
Table 3 shows the weighted variable means for each group,
and Table 4 gives the coefficients of separate determination.
Salinity, light transmittance, pH, productivity and chlorophyll
a are the important variables, with DO parallel to pH but to a
lesser extent. The vectors are plotted in Figure 10 for the
station groups.
Group 2 stations had the lowest mean temperature, produc-
tivity and chlorophyll a, and the highest salinity, DO, pH,
transparency and assimilation ratio. The high pH and DO do not
suggest inhibition, and in fact the phytoplankton may have been
stimulated. The zooplankton groups differ markedly from the
benthic groupings for the same period (section IVB).
June 1977.
Secondary waste treatment of TITP effluent began in April
1977 and may or may not have affected harbor station groupings,
but the populations appear to have been impacted. Certainly
the patterns show considerable overlap for Groups 1 and 2
(Figures 11, 12}. The TWT (Figure 13) shows that there were
less than half as many species present in June 1977 as were
present in December 1976 and only Group 3 (A1 outside the harbor)
has good populations. There may be normal drops in species in
the summer, perhaps due to predation.
In this period phytoplankton factors dominated the variables,
with only temperature and pH having minor roles, according to
the coefficients of separate determination. Group 3 was
separated by having the highest salinity, PH and transparency,
and the lowest productivity and chlorophyll a. Group 4 (station
A4), which rarely stands alone, was isolated in both zooplankton
and benthic analyses. For zooplankton it had the highest temper-
ature, productivity and chlorophyll a, and the lowest pH, trans-
parency and assimilation ratio. A bloom may have been just
getting underway (see section (IIC). Groups 1 and 2 were
intermediate in most measures, except for temperature, where
group 1 was lowest, and assimilation ratio, where group 2 was
highest. Figure 14 shows the important vectors and the station
groups.
-------
IVA 3
353
September 1911.
By September the zooplankton seemed to have mostly recovered
from the disturbance that had caused the low numbers of species
and organisms, except for the outfalls area, which had lost
most of its fauna. This suggests continuing impact locally
from effluent changes. This contrasts with the benthic fauna,
discussed in the next section, wherein the station groupings
continued to indicate more extensive abnormal separation.
The plankton station groups (Figures 15, 16) were more or
less arrayed concentrically from the outfall. However, the TWT
(Figure 17) shows that group 1 stations were low in numbers.
This included station AlO, which may have been affected by
residual oil, deposited after the Sansinena incident, and tends
to leach in warm weather. Groups 3 and 4 (the outermost
harbor and the sea buoy) were much richer.
The weighted means of variables measured are shown in
Table 7 and the coefficients in Table 8. Interestingly, all
variables were significant on one or more axes. These are
plotted in Figure 18. The outfall (Group 2) was highest in
weighted means for temperature, productivity and chlorophyll a,
and lowest in salinity, DO, pH and assimilation ratio. Group 1,
adjacent to the outfall station group, was second highest in
temperature, salinity, productivity and chlorophyll a, and
next lowest in DO, pH and assimilation ratio. Low assimilation
ratios in groups 1 and 2 suggest stress in the areas. The
sea buoy (group 4) was highest in DO, pH and assimilation ratio
and lowest in productivity and chlorophyll a. Group 3 stations
were colder than the sea station. The fluctuations in stabiliz-
ing secondary TITP effluent prior to diversion of cannery
wastes undoubtedly influenced the zooplankton to some extent
and the sessile benthic populations perhaps to a greater extent.
One cannery effluent was diverted to TITP in October 1977 and
the second by January 19 78. Were it not for the high coeffici-
ents for dissolved oxygen and assimilation ratios, the pattern
on the map might be considered as normal zonation.
December 1977.
The zooplankton patterns in December began to show an
increase in numbers of species and individuals, except for a
few anomalies (Figures 19, 20). Species numbers were not as
high as in December 1976, however. It is apparently a normal
winter pattern for the sea station to join outermost harbor
stations, as is true in group 1. Group 2, overlapping, is
distinct in having fewer species but with higher numbers
(TWT, Figure 21), whereas Group 3, station B8, appears to be
abnormally low in species. It was lowest in temperature and
in chlorophyll a, but highest in assimilation ratio and pH.
On the other hand, the outfalls area has merged with adjacent
stations (group 4) with increased species. Conditions in
-------
354
IVA 4
December might have represented an ideal situation, with
lower levels of cannery waste in combination with TITP second-
ary waste for the immediate effluent zone but the station B8
area seems to have suffered a retreat.
The weighted means (Table 9) showed a mixed pattern for
the groups, with group 1 having the highest means for salinity,
productivity and chlorophyll a and lowest for DO. However,
the coefficients (Table 10) showed that only temperature and
pH were significant physical variables, and the phytoplankton
variables were of greater importance. Group 4 sites had the
highest weighted mean temperature and lowest pH and assimilation
ratios. Thus a mixed pattern is achieved, based on physical
and biological variables. The vectors are plotted in Figure 11.
April 1978.
The April patterns for zooplankton showed some apparent
impact in the nearshore area, with the innermost station group-
ings mixed (Figures 23, 24). The TWT (Figure 25) shows a
reduction in species distribution and in numbers as compared
with December 1977, and the reverse might have been expected
when populations usually increase.
Weighted means and coefficients are given in Tables 11 and
12. All of the variables were important and separations were
made on four axes. Of the single station isolates, group 5
(station B9) had the highest weighted mean temperature, DO and
pH and the lowest transparency, productivity and chlorophyll a
as well as the second lowest assimilation ratio. It had large
populations of common zooplankton species, which may have
grazed the phytoplankton. Group 3 had the lowest mean temper-
atures and highest transparency and productivity. Group 4
overlapped, but had the lowest DO and pH and second lowest
temperature. Note that the waters were cooler closer to shore.
Group 1 had the highest assimilation ratio and lowest salinity,
and Group 2 had the lowest assimilation ratio and highest
salinity. The high dissolved oxygen and high coefficients for
phytoplankton suggest that a small, patchy bloom may have been
in progress. The vectors and station groups are shown in
Figure 26. The benthic patterns were also highly mixed in April.
It seems probable that the unusually heavy rains in January,
February and March (about 27 inches) were responsible and may
have led to the massive upset in the treatment plant in the
summer. The unstable patterns in Figure 23 may have been due
to variability in release of nutrients and in the control
measures instituted by TITP.
July 1978.
By midsummer the stations were again divided into 5 groups
(Figures 27, 28), but the separations were somewhat different.
The TOT (Figure 29) shows that species diversity was better than
-------
IVA 5
355
in the summer of 1977, although the summer appears to have
fewer species in the harbor when it is warmer than are there
in the winter. As was the case in April, station B9 stood alone,
this time as group 3; also the sea buoy station was separated,
as it often is in summer, and the outfall was separated, as
it is when nutrient levels are higher.
Group 2 (the sea buoy) had the lowest temperature and DO
(Table 13) but was intermediate on all other parameters. Group
3 had the highest weighted mean temperature and transparency,
and the lowest salinity, pH, productivity, chlorophyll a and
assimilation ratio. It would appear that water might have
pooled at B9 during the summer, where tidal water coming in
from the east meets the gyre. Group 5, the outfall, had the
highest DO, pH, productivity, chlorophyll a and assimilation
ratio, a very unusual circumstance. Groups 1 and 4 over-
lapped spatially and had intermediate values in the various
parameters. All parameters except light transmittance were
important according to the coefficients ( Table 14 ) . Vectors
are plotted in Figure 30.
September 1978.
The station pattern in September 1978 is very confused
(Figures 31, 32) and is probably indicative of the fact that
the high nutrient levels had again been terminated and
secondary treatment at TITP was brought back on line. The
"yo-yo" effects of waste treatment over the two-year period
have made populations transitory at best. In September, the
species list (TWT, Figure 33) was better than it was the
previous year, but oniy the stations on the periphery (groups
1, 2 and 5) appeared to have good populations. The split
between groups 3 and 4, with the outfall included in group 4,
is different from previous patterns.
Group 5 (B9 again was separated) was very different in
having much higher weighted mean productivity and chlorophyll
than had been seen for some time (Table 15) . It also had the
highest salinity, DO, pH and light transmittance but the
lowest assimilation ratio and temperature. Group 1 was second
lowest in temperature, salinity, and all three phytoplankton
measures. Group 2 (Al) had the lowest chlorophyll a, salinity,
DO, pH and transparency, an interesting reversal of patterns.
There had been about 0.5 in of rain the day before the samp-
ling so that salinities were low at the surface.
Groups 3 and 4 would probably have merged, except that
group 3 had the lowest productivity and highest assimilation
rates; and in all other measures the two groups were at inter-
mediate levels. The coefficients (Table 16) indicated that all
parameters measured except temperature and salinity were impor-
tant. This is also an unusual occurrence, but the lack of varia-
tion in the two parameters throughout the stations would explain
that.
-------
356
IVA 6
The vectors are plotted in Figure 34 for the station
groups. In this case group 5 is so different that the others
are arrayed on the opposite side, but good separation is
still accorded the groups.
LITERATURE CITED See Section VIA.
METHODS SECTION, DISCRIMINANT ANALYSIS See Section VIB.
-------
JL
¥tLOCITT 1CM.I
NORTH
SCALE OF FEtT
leoa o yooo
Source: McAnally. 1975
Figure 1
SURFACE CURRENT PATTERNS w
Base Test, Spring Tide Hour 6 ^
-------
		 ' —
vecocrrr scmx.f**
NOHTH
SCALE OF FEET
Source: MoAnolly. ^9/5
Figure 2
SURFACE CURRENT PATTERNS
Base Test, Spring Tide Hour 13
-------
IVA 9
359
,C11*
WILMINGTON
LONG BEACH
Pier J 7\
SAN
PEDRO
rVOUV
cjrouv
Harbors Environmental Projects
University of Southern California
Figure 3. Zooplankton Station Groups, December 1976.
Group i - A3, a^, as, A9, aii Group 3 - ai, a2, ai2
Group 2 - a7
-------
360
IVA 10
Figure 4.
TiiHHINAL ISLAND TREATMENT PLANT PLANKTON ** DECEMBER, 1976
DISTANCE
GR
200 160 120 80 HO 0
--4	4	4	4	4	i
		761201/A08
....1**1		76 1201/AQ9
I 1		761201/A03
!		76 1 201 /A 04
		76 1201/A 11
		761201/&07
		761201/A01
		76 1201/A 02
		761201/A 12
-------
IVA 11
361
Figure 5.
TERMINAL ISLAND TREATMENT PLANT PLANKTON ~* DECEMBEB, 1976
STATION GROUP
•
>
.75
TO
1
~
>
.50
TO
.75
-
>
.25
TO
.SO
•
>
.00
TO
.25
BLANK

.00


CLYIEMNjiSTH A HOSTBATA
ONCAEA MEDITEHBANEA
ONCAEIDAE ONCAEA
PSEUDOCA LANIDAE CLAUSOCALANUS
CLAUSOCALANUS FUECATUS
OITHONA PLUMIFERA
CORYCAEU S AM AZONICUS
SECYNOCEHA CLAU3I
EVA3NE SPINIFEBA
PENILIA AVIBOSTRIS
CALAMUS HELGOL A1IDICUS
CALOCALANUS STYLIREMIS
CLAUSOCALANUS MASTIGOPHOBUS
ACABIIA CLAU3I
CTENOCALANUS VANUS
IE MORA DI3CAUDA TA
TOBTANUS UlSCAUDATUS
ACABTIA TONSA
PABACALANUS PARVUS
LABI DOCZfiA TRISPINOSA
EVADNE NOBDdANNI
CORYCAEUS ANGLICUS
CITHONA SIMILIS
EUTERPINA ACUTI?RONS
PODON POLYP HEMOIDES
QITHONA OCULATA
1
2
3
7 7
7
1
6 6
3
6
1 1
1
1
2 2
2
2
y a
0
0
1 1
1
1
/ /
/
/
A A
A
A
0 0
£
0
9 4
7
2
7 7 7

7 7
6 6 6

b 6
1 1 1

1 1
2 2 2

2 2
0 0 0

3 0
1 1 1

1 1
/ / /

' /
AAA

\ A
0 0 1

3 1
3 J 1

1 2
£


~
*
*
—

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•

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•

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m

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+ *-4-

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-
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-------
362
IVA 12
AXIS 2

nemx>.
©
AXIS 1
-¥ pH
Chi 4
Figure 6. Plankton Station Groups and Axes, with Vectors
December 1976
-------
IVA 13
363
Table 1.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT PLANT PLANKTON ** DECEMBER. 1976
GROUPS	1	2	3
1.	TEMPERATURE	17.3326	17.4439	17.3885
2.	SALINITY	32.9399	32.9506	32.9366
3.	OXYGEN	6.5003	6.3217	6.9872
4.	PH	6.1379	8.1C14	6.1718
5.	PRODUCTIVITY	1.7753	1.5801	1.8108
6.	CHLOROPHYLL A	1.3736	1.6919	1.2268
7.	ASSIMILATION RATIO	1.3715	1.1716	1.4476
** ONE-WAY ANCVA FOR EACH VARIABLE * OF =	2. 24

VARIAQLE
F
1.
TEMPERATURE
C.06
2.
SAL I NI T Y
0 .0 1
3 .
OXYGEN
0.22
4 .
PH
0.24
5.
PRODUCTIVITY
0.02
6 .
CHLOROPHYLL A
0.31
7.
ASSIMILATION RATIO
C .06
-------
364
IVA 14
Table 2.
*~ LATENT ROOTS	AN! 0 SIGNIFICANCE TEST FOR EACH AXIS **
AXIS ROOT	%	CUW % CHI SGLARED	OF
1	8.863E-02	83.8	83.8	1.78	8
2	1.709E-02	16.2	100.0	0.36	6
COEFFICIENTS CF	SEPARATE DETERMINATION (X lOO/SL'MCARS VALUE)) **
TERMINAL ISLAND	TREATMENT PLANT PLANkTCN ** CECEMEEfi. 1976
AXES	1	2
I.
TEMPERATURE
6 .5
57 . 2
2.
SAL 1NITV
3 .5
C.9
3 .
OXYGFN
3.9
5.0
4 .
PH
19 .1
3.9
5.
PRODUCT 1VITY
1 .9
6.4
6 .
CHLOROPHYLL A
60 .9
23.7
7.
ASSIMILATION RATIO
4 .2
3.0
-------
IVA 15
365
;B6"
WILMINGTON
LONG BEACH
D1*
B3»
C2l
A7
SAN ^
PEDRO
B10*
Group 1
B9*
A3*
[A3'
81*
Group
AO*
Harbors Environmental Projects
University of Southern California
Figure 7. Plankton Station Groups, March 1977
Group 1 - A2, A3, A4, as, A12 Group 3 - A7, ah
Group 2 - ai, A9, aio
-------
366
IVA 16
FIGURE 8.
TEHHINAL ISLAND TREATMENT PLANT PLANKTON ** MABCH, 1977
DISTANCE
200 160 120 80 40 0	GRC
-•4	4	4	4	4	i
		77 0309/A02
I	1		77 0309/A03
I"". 		770309/A03 1
I "I			770309/A04
..I I-		77 0 309/A 12
. .			770309/A01
1				77 0309/A 10 2
-	1		77 0309/A09
I			77 0 309/A07
	1		77 0 309/A 11
-------
IVA 17
367
Figure 9.
TERMINAL ISLAND TREATMENT PLAN! PLANKTON * * flARCU, 1977
STATION GROUP
*
>
.75
TO
1
•f
>
.SO
TO
.75
-
>
.25
TO
.50
•
>
.00
TO
.25
BLANK

.00


CITHONA SIMILIS
OIIHONA SPINIRQ3TRIS
CQRYCAEUS AHAZONICUS
01TH ON A OCULATA
CITHONIJAS 0ITHONA
ONCAEIDAE ONCAEA
CITHONA PLUMIfERA
RHINCALA NUS NASUTUS
CANDACIIOAE CANJACIA
PENILI A AViEOSTHIS
C0RYCAEU5 ANGLICUS
EVAONE 3PINIFE3A
EVAJNE N OR JM AN NI
PGDON POLYPHEJiOIDES
ACARTIA TONSA
LABIDCCiSRA TRI3PIN0SA
PARACALANUS PARVUS
MICROSEIELLA NGRVEGICA
TEHORA UISCAJDATA
EUTERPINA ACU'II?RONS
ONCAEA V EN U ST A
7
7
v)
3
J
9
/ /
A A
0 0
6 4
7	7
7	7
3	0
3	J
U	U
9	9
/	/
A	A
|u	0
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7	7
7	7
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3	3
0	0
9	9
/	/ /
AAA
0	0 1
2	3 2
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-------
368
IVA 18
AXIS 2
Q
©
	I
AXIS 1
Figure 10.
Plankton Station Groupings and Axes, with Vectors
March 1977
-------
IVA 19
369
Table 3 *
WEIGHTED GROUP MEANS
TERMINAL ISLANO TREATMENT plant PLANkTQN ** MARCH, 1977
GROUPS
1
1.	TEMPERATURE	14.4329	I4.37ie	14.4980
2.	SALINITY	31.5397	31.5607	31.5462
3.	OXYGEN	S.4756	9.5403	9.3327
4.	PH	6.2092	0.2195	6.2026
5.	XTRANSMITTANCE	47.7434	51.9859	47.767S
6.	PRODUCTIVITY	I 2.ft0 3 3	1 1 . 7875	13.0199
7.	CHLOROPHYLL A	5.3071	4.6048	5.5775
e. ASSIMILATION RATIO	2.3053	2.3162	2.3093
** ONE-WAY ANOVA FOR EACH VARIABLE * OF =	2, 27
VARIABLE	F
1.	TPMPERATURE	C.C6
2.	SALINITY	0.02
3.	OXYGEN	C.02
4 .	PH	0.01
5.	fcTRANSMITTANCE	C.04
6.	PROCUCTIVITY	0.02
7.	CHLOROPHYLL A	C.09
a.	ASSIMILATION RATIO	C.CO
-------
370
IVA 20
Table 4.
** LATENT ROOTS AND SIGNIFICANCE TEST FOR EACH AXIS **
AXIS ROOT	%	CUM X CHI SQUARED	CF
1	2.256E-02	63.4	63.4	C.52	9
2	I.303E-02	36.6	100.C	0.3C	7
COEFFICIENTS OF SEPARATE DE TFA MI MAT I GN (X 1CO/SUM(A0S VALUE ) J
TERMINAL ISLAND TREATMENT PLANT PLANKTON ~* MARCH, 1977

AXES
1
2
1.
TEMPERATURE
8 .3
9.0
2.
SALIN I TV
0 .9
20. 3
3.
OXYGEN
0 .1
9.4
4 .
PH
14.4
1 .7
5.
XTRANSMITTANCE
10 .5
17.9
6.
PRODUCTIVITY
3 7.8
29.8
7.
CHLOROPHYLL A
26.4
6.5
8 •
ASSIMILATION RATIO
I .6
5.4
-------
IVA 21
371
WILMINGTON
LONG BEACH
C6»

j-Tojcr
\ D2*
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A 5.
B11*
•CI
At2<
B9»
AS'
A2"
B1*
A1»
A0»
Harbors Environmental Projects
University of Southern California
Figure 11. Plankton Station Group, June 1977
Group 1 - A7, as, A9, aio, A12 Group 3 - ai
Group 2 - A2, A3, ah	Group 4 - A4
-------
372
IVA 22
Figure 12.
TEBBIBAL ISLAND TBEATflENT PLANT PLANKTON ** JUNE, 1977
DISTANCE
200
160
-.4-
120
-•4-
80
-4-
40
-4-
0
4
GRO
77 0608/&09
770608/A10
770608/A12 1
770608/A08
7706 08/A07
7706 08/A03
770608/A11
770608/A02
7706 08/A01 3
7706 08/A04 4
-------
IVA 23
373
Figure 13.
TriEiSINAL ISLAND THEATMENC PLANT PLANKTON ** JUNE, 1977
STATION GROUP
•
>
.75
TO
I
~
>
.50
TO
.75
-
>
.25
TO
.50
•
>
.00
TO
.25
BLANK

.00


COHYCAEIDAE COHlfCAEUS
EVADNE SPINIFERA
COSYCAEUS AMAZONICUS
ACASIIA TONSA
LABIDOC^BA T8ISPINOSA
C0BYCASU3 ANGLICUS
PABACALANUS PAfiVUS
EVADNE N03DKANNI
PODON POLXPHEMOIDES
OITUONA SIfllLIS
BUTEHPIN A ACUTIfRONS
2 34
7
7
3
3
D
8
/
A
0
&
7 7
7 7

7 7
7 7

0 0
0 0

o b
6 6

a o
0 0

8 8
B 3

/ /
/ /

A A
A A

1 a
0 0

0 8
3 2

7 7 7
7
7
7 7 7
7
7
0 0 0
0
3
6 b fa
6
6
0 0 0
0
I)
8 8 8
8
8
/ / /
/
/
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A
A
0 1 0
1
D
9 2 7
1
1


*


*

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*


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• • •
-
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-------
374
IVA 24
AXIS 2
D
©
AR
Prod
Chi /I
G> ®
AXIS 1
Figure 14. Plankton Station Groups and Axes, with vectors
June 1977
-------
Table 5.
ighted group means
TERMINAL ISLAND TREATMENT PLANT PLANKTON ~* JUNE,
GROUPS	I	2
1.	TEMPERATURE	19.1095	19.2420
2.	SALINITY	33.6385	33.6359
3.	OXYGEN	7.6486	7.6606
4.	PH	7.962 3	7.9652
5.	XTRANSMITTANCE	61.9045	62.005a
6.	PRODUCTIVITY	12.8767	13.2863
7.	CHLOROPHYLL A	2.0327	2.0490
8.	ASSIMILATION RATIO	6.8422	7.0198
** ONE-WAY ANOVA FOR EACH VARIABLE * OF =	3,

VARI ABLE
r
1.
TEMPERATURE
0.03
2.
SAL INITY
o
•
o
3.
OXYGEN
0.02
4 .
PH
o
•
O
CD
5 .
XTRANSM1TTANCE
0.24
6*
PRODUCTIVITY
0.30
7.
CHLOROPHYLL A
0.18
a.
ASSIMILATION RATIO
o
•
o
•Ml
1977
3
19.£481
33.6506
7.5555
8.0031
6 7.6584
11.1094
1.7 7 13
6.a 130
4
19.3084
33.6451
7,5963
7.9438
54.7498	H
<
13.3312	>
2.1573	^
6-6267
GJ
^4
U1
-------
376
IVA 26
Table 6.
** LATENT ROOTS AND SIGNIFICANCE TEST FGR EACH AXIS	**
AXIS ROOT % CUM % CHI SCUAPED	DF
1	1.800E-01 76.3 76.3 5.46	10
2	S.068E-02 21.5 97.7 1.63	a
3	5.309E-O3 2.3 100.0 0.18	6
COEFFICIENTS OF SEPARATE CETEKMNATICN (X ICC/SUM(ABS VALUE)) **
TfchVlNAL ISL AND TREATMENT PLANT PLANKTON * * JUNE, 1977

AXES

1
2
3
1.
TEMPERATURE
1
.7
13.9
17.2
2 .
SALINITY
0
.7
1 .4
0.3
3.
OXYGEN'
1
.0
3.7
0.0
4 .
PH
2
.5
3. 7
12.0
5.
JjTRANSMI T TANCE
15
.6
5.8
7 • C
6.
PRODUCT I VITY
36
.6
23.2
2 .C
7.
CHLOROPHYLL a
40
. 1
29.5
39.2
8.
ASSIMILATION RATIO
1
.8
I 8.8
22.4
-------
IVA 27
377
C11»
WILMINGTON
LONG BEACH
C6»
C7/
84*
01-
C2l
SAN
PEDRO*
B10«
B11»
Group
A12
1A10«
A13»
A1"
AO*
Harbors Environmental Projects
University of Southern California
Figure 15. Plankton Station Groups, September 1977
Group i - A3, A4, aio, aii	Group 3 - A2, as, A9, ai2
Group z - A7	Group 4 - ai
-------
378
IVA 28
Figure 16.
TEEHIHAL ISLAND THE&TMEHr PLA.HT PLANKTON ~~ SEPTEMBER, 1977
DISTA NCE
200 160 120 SO <40 0
-.4	4	4	4	4	4
77 0914/A03
77091H/k10
77 0914/A04
770914/A11
77 0914/A07 2
77 0914/A02
77 091U/A09
3
77 0914/A08
77 09 It/A 12
770914/A01
4
-------
IVA 29
379
FIGURE 17.
TEHMINAL ISLAND TtUATME'NT PL AN I PLANKTON ** SEPTEMBER, 1977
STATION GROUP
12 3 4
•
>
.75
TO
1
+
>
.50
TO
.75
-
>
.25
TO
.50
•
>
.00
TO
.25
BLANK

.00


CLACJS0CALANJS EUECATUS
EVADNE SPINIFERA
PONT EL LOPS IS OCCID^NTALIS
EVADNE NUBDHANNI
PABACALANUS PASVUS
C0BYCAEU5 AMAZOtflCUS
COSY CAEIDAE COhYCAEUS
PEN!LIA AVI30STHIS
ONCAEA ilEDITE&EANEA
SHINCALA N'JS NASUTUS
ACAETIA TONS A
OITiiONA SIMILIS
ACAfil'I A CL A'JSi
01 Tit ON A OCULATA
COBYCAEUS ANGLICUS
LABIDCCEEA T2ISPIN0SA
PODON POLYP tfEiiO ID LS
ONCAEiDAE ONCAEA
TOUT ANUS DISCAUJATUS
7 7
7 7
0	$
3	9
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7
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tReproduced from
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-------
380
IVA 30
AXIS 2
3
DO
4	
Chi A
Prod.
4 Pti
4 AR
©
©
AXIS 1
Figure 18.
Plankton Station Groups and axes, with Vectors
September 1977
-------
Table 7.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT PLANT PLANKTON ** SEPTEMBER. 1977
GROUPS	12	3	4
1.	TEMPERATURE	17.9381	18.0017	17.8119	17.8349
2.	SALINITY	3 2.4201	32.3635	32.4378	32.4163
3.	OXYGEN	7.4469	7.0707	7.5585	8.0340
4.	PM	8.0922	8.C742	6.1139	8.1318
H
5.	PRODUCTIVITY	0.5713	0.6595	0.5485	0.4384
C. CHLOROPHYLL A	2.5482	3.0815	2.3267	1 .6912	U>
H
7. ASSIMILATION RATIO	0.2483	0.2163	0.2734	0.4345
+* ONE-WAY ANOVA FOR EACH VARIABLE + DF =	3. 36

VARIAULE
P
1.
TEMPERATURE
0.29
2.
SAL IN ITY
0.12
3.
OXYGEN
0.71
4 .
PH
0.62
5.
PRODUCT IVITY
0.18
6 .
CHLOROPHYLL A
0 .83
7.
ASSIMILATION RATIO
0.50
-------
382
IVA 32
Table 8.
*~ latent roots ano significance test for each axis	**
AXIS ROOT X CUM X CHI SQUARED	OF
1	3.591E-01 74.3 74.3 10.20	9
2	1.C03E-0I 20.7 95.0 3.20	7
3	2.403E—02 5.0 100.0 0.80	5
COEFFICIENTS OF SEPARATE DETERMINATION (X 100/SUMCAQS VALUE)) ~~
TERMINAL ISLANO TREATMENT PLANT PLANKTON *» SEPTEMBER. 1977

AXES
1
2
3
1.
TEMPERATURE
2 .5
7.9
24.6
2.
SALINITY
6.3
18.4
8.1
3.
OXYGEN
37 .2
14.C
39.2
4.
PH
13.4
o.o
IS .0
5.
PRODUCTIVITY
4.8
10.7
1.8
6 .
CHLOROPHYLL A
27.1
3 . 1
10.9
7.
ASSIMILATION RATIO
0.6
45.9
0.4
-------
IVA 33
383
-B6'
CiT«,
WILMINGTON
LONG BEACH
BSi
C7/
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. dC2%
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A7
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J6»
A12'
A14.
Ac
A17»
Grouv 1
A13
B1«

A1»
AO*
Harbors Environmental Projects
University of Southern California
Figure 19.	Plankton Station Groups, December 1977
Group 1	-	ai, a2, A3, A12, ai3, Aie, A17
Group 2	- as, Ai4
Group 3 - bs
Group 4	-	A4, at, ais
-------
384	IVA 3 4
Figure 20.
IEBMINAL ISLAND TREATMENT PLANT PLANKTON ** DECEMBER/ 1977
DISTANCE
200 160 120 80 40 0	GR0UF
--4	4	4	4	4	4
		771206/&03
I""I		771206/A16
..I		771206/A 01
I :		771206/A09
		771206/A02 1
.1 1-2".		77 1 206/A 12
Zl I		77 1206/A 17
l"l'.		77 1 206/A 13
	I ~Z 		77 1 206/A08
I J I		77 1 206/A 1
' r		771206/B 08 3
		77 1206/A04
I	I 1		77 1206/A 15 4
1		771206/A07
-------
IVA 35
385
Figure 21.
IEBIUHaL ISLAND TaiitTiUNT PLANT PLANKTCN ~ ~ U£.C£i15Efc, 1977
STATION GROUP
23 4
•
>
.75
TO
1
*
>
.50
TO
.75
-
>
. 2S
TO
.50
•
>
.00
TO
.25
BLANK

.00


ACAKT1A CLAUSI
CLAUSOCALABUS PARAPERG2N5
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TORT ANiJS jlSCAUDATUS
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-------
386
IVA 36
AXIS 2
©
<_pi
AS
©
©
AXIS 1
AXIS 3

pg
©
©
©
AXIS 1
Figure 22. Plankton Station Groups and Axes with Vectors
December 1977
-------
Table 9.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT PLANT PLANKTON »* DECEMBER, 1977
GROUPS	12	3	4
1.
TEMPERATURE
I 7.9180
17.9246
17.8985
17.9250
2.
SAL INITY
34.2526
34.2464
34.2382
34.2074
3.
OXYGEN
7.4464
7.4929
7.5351
7.5711
4 *
PH
6.1674
8.1738
8.1810
8. 1524
5.
PRODUCTIVITY
1.2923
1.2060
1.2461
1 .2131
6.
CHLOROPHYLL A
I.2235
1 .1333
1.1176
1.2058
7.
ASSIMILATION RATIO
1.2235
1 .2351
1.3383
1.1454
~* CNE-WAY ANOVA FOR EACH VARIABLE ~ OF =	3. 52
VARIABLE	F
1.	TEMPERATURE	0,02
2.	SALINITY	0.01
3.	OXYGEN	0.01
4.	PH	O.OS
5.	PRODUCTIVITY	0.02
6.	CHLOROPHYLL A	0.02
7.	ASSIMILATION RATIO	0.09
u>
00
^sj
-------
388
IVA 38
Table 10.
** LATENT ROOTS	AND SIGNIFI
AXIS ROOT	%
1	2•fi 7 OE— 02	69.2
2	ti•751E— 03	21.1
3	4.032E-03	9.7
ANCE TEST FCH EACH AXIS	**
CUM * CHI SQUARED	DF
69.2	1.40	9
90.3	0.43	7
100.0 0.20	5
VALUE)) **
1577
1.
TEMPERATURE
5 .3
3.2
27.2
2.
SAL INITY
Z .4
7.5
6. 1
3.
OXYGEN
1 .6
6.3
4.0
4 .
PH
17.1
3.2
2.9
5.
PRODUC T I V I TY
1 2 .6
26. 1
25.4
6.
CHLOROPHYLL A
25 . 1
20.3
23.9
7.
ASSIMILATION RATIO
00
•
in
25.4
10.5
COEFFICIENTS OF SEPARATE DETERMINATION (X 100/SUM(A6S
TERMINAL ISLAND TREATMENT PLANT PLANKTON ** DECEMBER.
AXES	1	2	3
-------
IVA 39
389
;B6'
WILMINGTON
LONG BEACH
Co»
&£• K-yC10«
C7/
D2«

,C3*
B4.
B2« J;
Pier:J_:\
D1»
B3*
A7
SAN
PEDRO'
B10»
FA16'
A12'
AU.
Group 1
A17»
A2»
,10«
A13
B1*
fO<
A1«
AO*
Harbors Environmental Projects
University of Southern California
Figure 23. Plankton Station Groups, April 1978
Group i - ai, A2, ai2, Ai4, ais, ai7, bs
Group z - aio	Group 4 - A3, a7, as, A9
Group 3 - A4, ah, ai6 Group 5 - B9
-------
390
IVA 40
Figure 24.
TEfiHINAL ISLAND TBEATHENT PLANT PLANKTON ~» APfilL, 1978
DISTANCE
200
--4.
160
-.4-
120
• -4 ¦
80
.4-
40
-4.
0
4
GROUP
780405/A02
780405/114
780405/A01
780405/A12
780405/A15
780405/A17
78 0405/B08
78 0419/A 10
780405/A04
78 0405/A11
780405/A16
780405/A08
780405/A09
780405/A03
780405/A07
780405/B09 5
-------
IVA 41
391
Figure 25.
TERMINAL ISLAND TREATMENT PLANT PLANKTON ** APRIL, 1978
STATION GROUP
2 3
•
>
.75
TO
1
+
>
.50
TO
.75
-
>
.25
TO
.50
.
>
.00
TO
.25
BLANK

.00


ACABTIA DANAE
CALOCALANOS STYLIREMIS
CAMOACXIOAE CANDACIA
COBYCAEiiS GIESBRECHTI
LABIDOCEHA JOLLAE
PODOH POLYPHEMOlDES
PSEUDO-PELTIDIIDAE CLYTEMNESXR
CTENOCAL ANUS VANUS
ONCAEIDA E OHCAEA
IHINCALANUS NASUTOS
OITHONA PLUMIFERA
PSEODOCALAHIDAE CLAUSOCALANUS
TEMOBA DISCAODATA
ACABTIA CLAOSI
COBYCAEUS AMAZONICUS
EVADNE SPINIFEBA
EVADNE NOBDMANNI
PARACALANUS PABVUS
ACABTIA XONSA
E0TERPINA ACOTIFBONS
LABIDOCEBA TRISPINOSA
COBYCAEUS ANGLICUS
PEMILIA AVIBOSTRIS
COBYCAEIDAE COBYCAEUS
01THONIDAE OITHONA
CLAOSOCALANUS FUBCATOS
OITHONA OCDLATA
CALANUS HELGOLANDICOS
IOBTANUS DISCAUDATUS
CITHONA SIMILIS
OHCAEA VENU5TA
7	7 7
8	8 8
0	0 Q
4	4 4
0	0 0
5	5 5
/	/ /
AAA
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-------
392
IVA 42
	
AXIS 3
FIGURE
26.
Plankton Station Groups
April 1978
and Axes with Vectors
-------
Table 11.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT
PLANT PLANKTON ** APRIL. 1978
GROUPS
1.	TEMPERATURE	17.3622	17.3739	17.2925	17.3472	17.3920
2.	SALINITY	34.3463	34.5700	34.5251	34.5456	34.5262
3.	OXYGEN	10.7981	10.7Q45	10.7066	10.6503	11.2950
4»	PH	7.8899	7.8626	7.8623	7.861 1	7.9024
5.	XTRANSMlTTANCE	47.9610	48.6545	51.1607	49.3359	43.7243
6.	PRODUCTIVITY	4.0545	3.7400	4.0674	3.8385	3.5790
7.	CHLOROPHYLL A	1.7964	1.8225	1.6993	1.7257	1.6744
8.	ASSIMILATION RATIO	2.5422	2.3215	2.5378	2.4596	2.3914
H
$
U>
** ONE-WAY ANQVA FOR EACH VARIABLE * DF -	75

variable
F
1.
TEMPERATURE
C.05
2.
SAL INITY
0.03
3.
OXYGEN
0.06
4 .
PH
0.03
5.
XTRANSMITTANCE
C.06
6.
PRODUCTIVITY
0.03
7.
CHLOROPHYLL A
0.03
8.
ASSIMILATION RATIO
0.04
-------
394
IVA 44
Table 12.
~* LATENT ROOTS AND SIGNIFICANCE TEST FDR EACH AXIS *»
AXIS ROOT	%	CUM X	CHI SQUARED	OF
1	3•236£—02	56.7	56.7	2.31	il
2	1.382E-02	24.2	8 1.0	C.99	9
3	8.338E-03	14.6	9S.6	C.60	7
4	2.516E-03	4.4	100.0	0.18	S
COEFFICIENT S OF SEPARATE DETERMINATION (X 1CC/5UM(AOS VALUE)) **
TERMINAL ISLAND TREATMENT PLANT' PLANKTON ** APRIL. 1978
AXES	12	3	4
1.
TEMPERATURE
1 1 .2
6.8
7.6
0.4
2.
SAL INITY
1 .2
4.6
56.7
2.8
3.
OXYGEN
19 .6
7.1
4.0
C.9
4 .
PH
7.0
0 .6
1 .6
5 1.3
5 .
XTRANSMITTANCE
34 .6
0.3
3.8
1.5
6.
PRODUCTIVITY
7.1
19.9
0.7
20.2
7.
CHLOROPHYLL A
2.1
35.3
4.4
6.6
8.
ASSIMILATION RATIO
1Q .3
25.5
19.2
16.3
-------
IVA 45
395
;B6;
WILMINGTON
LONG BEACH
ce» fer
C10*
D2*
03*
C3«
B4*
B2*
Pia'f J \
D1«
B3«
C2I
A6*
A7
SAN •>;:
PEDRO;
B10*
A12'
,3»
A#'
A17' Group 1
7k2*
A10*
B1*
A1»
AO*
Harbors Environmental Projects
University of Southern California
Figure 27. Plankton Station Groups, July 1978
Group i - A2, aa, A9, Ai2, ai3, ai4, ai7, bs
Group 2 - ai	Group * - A3, as, aio, ais, aic
Group 3 - B9	Group 5 - A7
-------
396
IVA 46
Figure 28.
TEBaiKAL ISLAND TfiEATHENT PLAHT PLANKTON ~* JOLI, 1978
DISTANCE
200
...J.
160
--4-
120
--•I-
80
-4-
uo
0
4
GRO
"780705/A 14
780705/A17
780705/A02
780705/A09
780705/A13
78 0705/A12
780705/A04
780705/B08
780705/A01
780705/B09
780705/A15
780719/A10
780705/A16
780705/A03
780705/A08
2
3
780705/A07 5
-------
IVA 47	397
Figure 29.
TERMINAL ISLAND TREATMENT PLANT PLANKTON ~* JULY, 1978
STATION GROUP
2 3 4
•
>
.75
TO
i
~
>
.90
TO
.75
-
>
.25
TO
.50
•
>
.00
TO
.25
BLANK

.00


CALOCALANUS STYLIREMIS
PS EUDGCALAN ID AE CLAUSOCALANUS
EVADNE NOHDdANNI
BHINCALANUS NASUTUS
CORYCAEUS ANGLiCUS
P&HACALANUS PAEVUS
ACARTIA TONSA
CITHONIDAE OITHGNA
CALANUS HELSOLAMDICUS
PODON POLYP maolDtuS
IORIANUS DISCAUJATUS
LABIDOC&HA TRISPINOSA
GITHONA OCULATA
ONCAEIDAE OH CAE A
OITHONA SI HI LIS
CALANUS HINOR
CEPHALOPODA OCTOPODA
PENILIA AVIHOSTHIS
7 7 7 7

7
7 7
ct a a a

8
8 8
0 0 0 J

i)
0 0
7 7 7 7

7
7 7
j o o a

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5 5 5 5

5
9 5
/ / / /

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0

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7

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5 5 5 5
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Reproduced from
best available copy.
-------
398
TV A 4 8
Trans.
AXIS 3
©
3
©	
AXIS 1
Figure 30. Plankton Station Groups and Axes with Vectors
July 197a
-------
Table 13.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT PLANT PLANKTON *~ JULY ,
GROUPS	1	2
1.
TEMPERATURE
16.3575
16.2665
2 •
SAL INI TY
3 0.7729
30.7660
3*
OXYGEN
8.4 123
8.3522
4 .
PH
8.3675
8.4052
5.
XTRANSMITTANCE
69. eeae
69.0129
6.
PRODUCTIVITY
3.9408
3.9742
7.
CHLOROPHYLL A
4.4884
4.4040
a.
ASSIMILATION RATIO
0.854 5
0.8713
** ONE-WAY ANOVA FOR EACH VARIABLE ~ OF =	4.

VARIABLE
F
i .
TEMPERATURE
o
•
o
2.
SAL IN I T Y
o
o
3.
OXYGEN
o
•
o
4 .
PH
C .04
5.
XTRANSMITTANCE
C. 1 I
6.
PRODUCT I VITY
0.2 1
7 .
CHLOROPHYLL A
0.16
a.
ASSIMILATION RATIO
0.02
1978
3
4
5
16.4917
16.3300
16.2695
30.7453
JO.755 7
30.7568
8.4349
8.5 143
8.5526
8.3626
8.4445
8.49 13
70 .2 044
69.2063
67.0388
3.9073
4.2457
4.9648
4 .3076
4 .6408
5.0495
0.852 1
0.8648
0.9073
LtJ
V0
KD
-------
400
IVA 50
Table 14.
** LATENT ROOTS AND SIGNIFICANCE TEST FOR EACH AXIS **
AXIS ROOT	X	CUM X	CHI SQUARED	OF
1	4.186E-02	72.7	72.7	2.97	11
2	1.26 IE —02	21 .9	94.5	C.91	9
3	1.819E-03	3.2	97.7	C.13	7
4	1.324C-03	2.3	100.0	C.10	5
COEFFICIENTS OF SEPARATE GETERHINATfON (X 100/SUM(A6S VALUE))
! MINAL ISLAND TREATMENT
PLANT
PL ANKTON
»* JULY,
1978

AXES
1
2
3
4
1 .
TEMPERATURE
1 0 .7
57.7
1.6
3.6
2 .
SAL IN ITY
0 . 1
16.6
16.6
1 . 4
3 .
OXYGEN
3 .2
17.9
1 .4
36.8
4 .
PH
IC .0
C. 5
5.5
2 1.5
5.
XTRANSMITTANCE
7 .2
0.7
5.0
1.3
6 .
PRODUC TI V ITY
62 .5
2.0
6.2
34.4
7 .
CHLCRGPHYLL A
0 .6
4.6
59.0
C • 4
6.
ASSIMILATION RATIO
5 .a
0.1
4.6
c.a
-------
IVA 51
401
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University of Southern California
Figure	31. Plankton Station Groups, September 1978
Group	i -	as, bb	Group 4 - A2, A4, A7, A9, aio, ai3,
Group	z -	ai, A3	Ai4, Ai6, ai7
Group	3 -	ah, ai2, ais	Group 5-39
-------
402	IVA 52
Figure 32.
TERMINAL ISLAND TREATMENT PLANT PLANKTON ~~ SEPTEMBER, 1978
01 STANCE
GROL
200	160	120	80	40	O
	..}	I.».	4	4	*
780906/A08
1
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780906/A01
780906/A03

780906/A11
• ••••
780906/A15

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780906/A16
• • •
780906/A17
•• •••••••••
780906/A14
• •
•• •••••••••
78O906/A0 2
••••••
780906/A13
•
780906/A04
780906/A09
780906/A07
7B0920/A10
780906/B09 5
-------
IVA 53
403
Figure 33.
TEBttlNAL ISLAND TREATMENT PLANT PLANKTON ** SEPTEMBER, 1978
STATION GROUP
1 2 3
•
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TO
1
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ACABTIA DANAE
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PODON POLY? HEMOIDES
EVADNE NORD MANNX
LABIDOCEHA TRISPINOSA
TORTAHUS DISCAUDATUS
CLAUSOCALAHUS fURCATUS
PSEUDOCALANIDAE CLAUSOCALANUS
PENILIA AVIROSTBIS
OITHONA SI3ILIS
ONCAEIDAE ONCAEA
CTEN0CALANU5 VAHUS
EUTERPINA ACUTIfROSS
CALOCALANUS STYLIREMIS
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7
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-------
404
AXIS 2
©
©
©
3
IVA 54
Prod.
Chi A
©
AXIS 1
Figure
39.
Plankton Station Groups
September 1978
and Axes with Vectors
-------
Table 15.
WEIGHTED GROUP MEANS
!M IN
IAL ISLAND TREATMENT
PLANT PLANK10N
*~ SEPTEMBER
, lS7e



GROUPS
1
2
3
4
5
1 •
PRODUC TIVI TV
t 3* 58 21
13. 7260
1 5 .3931
14.8704
23.3209
2.
CHLOROPHYLL A
3.7622
3.7423
3.3 9 53
3.9099
7.5790
3.
ASSIMILATION RATIO
3.6650
3.7624
4 .0033
3.0597
3.2790
4.
TEMPERATURE
I 8. 4 4 25
18.4074
1 6.4770
18.4761
18.3 199
5.
SAL IN ITY
30.5036
30.5803
30.5929
30.5671
30.6425
6 .
OXYGEN
6.3704
6.3445
6 .3704
6.3597
6.6032
7 .
PH
Q.535I
8.5203
8.5419
fl.5346
8.6163
a.
XTRANSMIT TANCE
64.9639
62.9756
64.5029
64.1015
68.7297
** ONE-WAY ANOVA FOR EACH VARIABLE * DF =	4, 00

VARIable
F
1 .
PRODUCT 1VITY
0.58
2.
CHLOROPHYLL A
1 .35
3.
ASSlMILATION RATIO
0.17
4 .
TEMPERATURE
0.03
5.
SALINITY
0.03
t .
OXYCfcN
0*
o
•
o
7.
PH
0.08
e.
%TRANSMITTANCE
0 .04
-------
406
IVA 56
Table 16.
** LATENT ROOTS AND SIGNIFICANCE TEST FOR EACH AXIS *+
AXIS ROOT	X	CUM %	CHl SQUARED	OF
1	3.665E-01	92.9	92.9	24.20	11
2	2.133E-02	5.4	98.3	1.64	9
3	6.11OE—03	1.5	99.9	0.47	7
4	4.506E-04	0.1	1CQ.0	0.03	5
COEFFICIENTS OF SEPARATE DETERMINATION (X 100/SUM(A8S VALUE)) **
TERMINAL ISLAND TREATMENT PLANT PLANKTON ** SEPTEMBER. 197S

AXES
1
2
3
4
1.
product I V Ity
26 .7
26. 1
27.9
1 0.9
2.
CHLOROPHYLL A
60 . 1
0.0
19.1
19.6
3.
ASSIMILATION RATIO
5 .1
61.4
3. 1
39. 7
4.
TEMPERATURE
2 .9
4.6
7 . 1
3.0
5 .
SALINITY
0 .4
1 . S
4.0
5.0
e.
OXYGEN
0 .3
4.3
6.0
11.6
7.
PH
4 .0
I .7
11.6
o.a
8.
XTRANSMITTANCe
0 .5
0.4
2 1.3
9.5
-------
IVB
407
WEIGHTED DISCRIMINANT ANALYSIS OF BENTHIC DATA
Computer programs developed for ecological analysis in
Los Angeles Harbor were first used on the 1973-74 benthic data
(AHF, 1976). In that report, 34 benthic stations were sampled
quarterly in the two harbors. Discriminant analysis showed
a pattern (1976, Figure 6.3) based on annual data of inner
blind-end slips, main channel, outer harbor and polluted zone
faunal separations. Henry (1976) and Soule and Oguri (1973)
showed that seasonality existed in benthic faunal composition,
which had previously been underestimated or disregarded. There-
fore, the quarterly benthic data from December 1976 have been
analyzed in the present report.
RESULTS
December 1976.
Twelve A stations were sampled in December 1976 in outer
Los Angeles Harbor. The station groupings (Figures 1 and 2)
could very well be viewed as being separated by depth and dis-
tance from the shorelines, in fairly typical winter season
patterns.
The Two-Way Table (TWT, Figure 3) provides information
that the number of species was very low at A4 and A7, whereas
A1 had a number of species not seen at other stations. The
weighted group means are presented in Table 1, and the coeffi-
cients of separate determination are given in Table 2. The
coefficients show that the physical variables did affect
separation, primarily by salinity, and less so by dissolved
oxygen, temperature and depth, but the biological variables
for productivity and assimilation ratio were also of prime
importance.
Group 1 stations had the highest weighted means for pro-
ductivity and assimilation ratio and the highest salinity and
dissolved oxygen. Group 2 stations had the next highest.
Group 4, the outfalls, had the lowest weighted mean temper-
ature, lowest salinity, DO and assimilation ratio, and is
the shallowest as well. Group 3 (the sea buoy) had the
highest temperature and lowest productivity. The coefficients
showed that Axis 1 contained 66.9% of the information, but
Axes 2 and 3 were also effective in separation. Figure 4
shows the separation of the groups according to vectors for
the particular parameters which could be diagrammed on the
axes. Usually only those with coefficients of ten or above
are indicated on the figure of the vectors.
-------
408
IVB 2
March 1977.
Groupings for this period appeared to be somewhat similar
to the December 1976 separations, ranging from shoreline to
outside the harbor. However, Groups 1 and 3 overlapped, due
to the inclusion of station A4 with outer harbor Group 3
(Figures 5 and 6). The isolation of Group 2 {station Al) is
not unexpected, since it had the lowest weighted mean temper-
ature as well as the greatest depth. It also had the lowest
chlorophyll a values (Table 3).
Examination of the coefficients of separate determination
show that pH values were most important on Axis 1 (Table 4)
and helped to isolate the sewer outfall (group 4, station A7).
Group 1 was the highest and group 4 the lowest in salinity,
dissolved oxygen, pH, primary productivity and assimilation
ratio. The outfall (group 4) was highest in temperature and
chlorophyll a values, the latter also having a high coefficient
on Axis 1. Diversity was very poor at the outfall area, and large
gaps in benthic species can be seen in Grouc 3 fauna in the TOT
(Figure 7). Groups 2 and 3 varied in the intermediate rankings
of the various parameters. Group 3 was very close to group
in temperature, DO and productivity, which may account for
the overlap. The separation is so pronounced for the outfall
area that it masks the other trends. If the intent of the
analysis were not to characterize the effects of the outfalls,
but rather to describe the characteristics of the other sta-
tions, the analysis would have been repeated without the A7
data in order to create maximal separation of the other
stations (Figure 8). Although Axis 1 contains 99% of the
separation values, temperature and productivity dominate
Axes 2 and 3 (Table 4). However, the vectors of parameters
measured cannot be plotted on those axes alone.
The dissolved oxygen and phytoplankton measurements sug-
gest that a bloom was in progress. This is confirmed by
comparison with December data (Table 1) and with section IIC
on primary productivity. The weighted means of the phyto-
plankton variables (Table 3) show pronounced separations,
particularly between Groups 1 and 4, which are in fact nearest
physically.
The rainfall for the winter of 1976-77 was considerably
below normal and may have affected the harbor life cycles in
unknown ways.
June 1977
The beginning of secondary waste treatment in April 1977
may or may not have affected the benthic populations in the
outer harbor. However, in section HE, Figures 6 and 7 showed
that the number of species dropped sharply by June, except at
-------
IVB 3
409
A2 and A7 which were already low, while the numbers of organ-
isms increased except at Al and A9.
Figures 9 and 10 show the isolation into separate groups
of several stations, except for those clustered in the central
gyre of the outer harbor (Group 1). That group seems to be
the richest (TWT, Figure 11). The outfall area had a few more
species than before, and Al no longer stood alone, as it
frequently does in the winter.
The weighted group means (Table 5) showed that the outfalls
area (Group 5) was highest in temperature, productivity and
chlorophyll a and lowest in salinity, dissolved oxygen, pH and
assimilation ratio. The pH, salinity and oxygen levels were
not low enough to be considered limiting, and their relatively
low coefficients of separate determination (Table 6) show that
these were not important factors. Depth and temperature were
much more important, since they are represented by high coeffi-
cients. Chlorophyll a values were by far the most important
of the biological values measured.
Group 1 stations were highest in salinity, pH and assimi-
lation ratio and lowest in productivity. The values for the
ther groups were mixed in ranking. Group 2 (station A9) was
lowest in productivity, which may reflect the lingering effects
of the Sarts inena tanker explosion and Bunker C spill that
occurred six months earlier at that location.
September 1977.
The stations for September 1977 were divided into five
groups with some overlap and spatial separation (Figures 13
and 14). However, the most extreme contrast was between
Group 5, the outfalls area, and Group 1, the outermost harbor
stations. Groups 2, 3 and 4 were so similar to Group 1, in
contrast to the outfalls, that separation was difficult. The
weighted means (Table 7) and coefficients of separate deter-
mination (Table 8) show that the phytoplankton measurements
were very low, and only assimilation ratio was an important
coefficient in separation on Axis 2. The outfall was poor in
species also (TWT, Figure 15). Weighted means of Group 5
showed that the outfalls area had the highest pH and salinity
and least depth; it also had the highest productivity and
chlorophyll a and the lowest assimilation ratio. Vectors are
shown in Figure 16. The variability in salinity from month
to month at the outfall at times has been due to rainfall
runoff (there were 2+ inches in mid-August 1977), or to the
amount of cannery wastes processed. Cannery wastes may be
highly saline because of the freezing brines from boat holds;
this was formerly diluted during processing. The salinity
coefficient was one of the most important coefficients on
Axis 1, along with depth and then pH. The salinities were not
low enough to have great impact, but the rain in August could
-------
410
IVB 4
have favored opportunistic species such as Cavitella a agitata.
Salinities were about 1.8 ppt lower than in July 19 77.
It is surprising to see such low phytoplankton measure-
ments over the entire outer harbor, when fall peaks would
normally have been building, perhaps giving an indication of
the inhibitory impact of alterations in the character of the
TITP and cannery wastes. The benthic species and numbers
appear to be reudced as well.
January 1978. Harbor grouping appeared to be fairly typical
of winter, with concentric outer harbor bands and with the sea
buoy (group 3) isolated. By January 1978 both cannery effluent
lines had been diverted to TITP. Some biostimulation may have
been occurring in the A9 area of the Sansinena (Figures 17 and
18) at Group 1 stations; the TWT (Figure 19) also suggests this.
However, the weighted means (Table 9) show Group 1 as highest
in temperature, salinity, DO and pH. Phytoplankton measures
were all low. The coefficients (Table 10) showed that phyto-
plankton measures were not as important as the physical
measures, except for assimilation ratio. Group 5, the outfall
area, was isolated by having the lowest weighted mean salinity,
temperature and DO (Table 9) even though salinities were quite
low at all stations.
A heavy rainfall of 1.5i in. was in progress during actual
benthic sampling, lowering all salinity values in the harbor.
Heavy rainfall runoff appears to lower DO in the harbors, as
was documented in Marina del Rey for storm flows (Soule and
Oguri, 1976). The pH weighted mean was lowest at the sea
station Al (Group 3), but productivity and chlorophyll were
higher there. In Table 10 the coefficients of separate deter-
mination especially emphasize the physical parameters of pH
(73.9% on Axis 2), oxygen (56.5% on Axis 1), temperature,
and salinity.
The impacts of rainfall have not been well documented,
but would not be expected to affect benthic organisms except,
for example, at the outfalls area (group 5) where storm
runoff goes through TITP. Stone and Reish (19 65) documented
the effects of rainfall on benthic organisms at the mouth of
the Los Angeles River, and Soule and Oguri (unpublished data)
noted the removal of benthic organisms and recolonization
in Dominguez Channel (Shell Corp. Pipeline Crossing EIR).
Opportunistic species recolonize in a matter of a few weeks
in the area where existing sediments were carried away and
new ones deposited (Soule and Oguri, 1976). Such an area
may support only a few species that are euryhaline and have
year-round reproductive cycles, such as Capitella This is
even more the case where the character of the effluent is
varying widely, as it was with all the alterations in waste
disposal treatment in 1977-78 (see section HE). Prior years
had not shown such extreme domination in 1973-74 (AHF, 1976).
-------
IVB 5
411
April 1978.
The trends in the harbor in April 1978 were difficult to
interpret, with only Group 3, nearest the outfall, clearly
separated (Figures 21 and 22). The other two groups overlapped
extensively. Group 3 (stations A4 and A7) had the highest
weighted mean temperatures and the lowest dissolved oxygen and
light transmittance. However, oxygen readings were high,
almost as they are at the beginning of a bloom, but more prob-
ably due to mixing from storms and high tides. While group 3
had the highest phytoplankton measurements, all chlorophyll a
and productivity readings were quite low for the season. There
is usually a spring peak, but in April 1978 the weighted mean
values (Table 11) were more like the winter lows of December
1976 and January 1978 (Tables 1 and 9) than they resembled
the spring 1977 values. The unusual rainfall of about 27
inches (unofficial) between January and April probably was
responsible. Normal rainfall is about 14 in. a year.
Chlorination was carried on at TITP from the end of March
through August 1978, and this might have been partly responsible
for the very low assimilation ratio, which indicates stress.
However, the imemdiate outfalls area seemed to have slightly
better levels of chlorophyll a than the rest of the harbor,
suggesting that the drop was not caused by TITP but by natural
weather conditions. It appears that the entire coastal area
was involved, as shown by party boat fish catches (IIA), which
were down outside the harbor even though there were actually
only four days of measurable rain in April in contrast to
about 10 days in January, and 11 each in February and March.
The benthos, phytoplankton, zooplankton and fish inside the
harbor were all lower than would be expected, and no spring
reproductive surge could be seen. Sea temperatures had
remained quite warm through the spring until early summer,
with a late cooling period. Thus the change in temperature
necessary as a reproductive cue in many organisms (Vernberg
et al. 19 77) was out of phase. The combination of factors,
or some parameters not measured, were responsible for the
widespread cumulative decrease in organisms. Salinities
were higher in April in the harbor than in January, in spite
of the rains, suggesting an influx of coastal water, and pH
was lower than usual. All of the coefficients (Table 12) of
variables measured were moderately important except for
chlorophyll a, but no single coefficient was very high except
for pH; it is usually 8 or above in the harbor except in stressed
areas. Group 2 strongly suggests the gyre water, with an
influx of water with the highest salinity, oxygen and trans-
parency into the harbor. Group 1 is probably the residual
water of lower salinity that became the center of the gyre
or pooled.
-------
412
IVB 6
July 1978.
In contrast to the obvious impact of natural conditions
in April, July patterns clearly showed the effects of the
breakdown at TITP, but not necessarily negative ones. The
harbor showed one large station group overlapping another
and several single station isolates (Figures 25, 26). There
was a reduction in species (Figure 11) at station 4 (Group 4)
over June 1977, but an increase at A7 (Group 5) in July 1978
(TWT, Figure 27). Over all, species numbers were up over the
previous July.
While benthic organisms would not seem to be as affected
as plankton by the variabilities in the water column, it
appears that the opposite is the case. The TITP breakdown in
July increased the nutrients and bacteria in the water column.
Salinities and temperature were lower than they were in April
and were significant (Tables 13, 14), but dissolved oxygen was
the most important variable according to the coefficients.
Dissolved oxygen was below 5 ppm in Group 3 in an area where
floating solids had collected, and Group 1, a large area, was
next lowest. The outfall itself (Group 5, A7) had the highest
DO, but this was only slightly higher than the others, and all
were below mean 6 ppm. Station 4 was highest in weighted mean
phytoplankton measures and Group 1 was the lowest, but these
were all very low for summer. The increase in species and
numbers at A7 (Group 5) is unexpected, since sudden changes
in effluent character might have been inhibitory. High DO and
pH may have been due to control efforts. The group 4 isolate
is peculiar in that the three phytoplankton measurements, all
with significant coefficients, were not normally proportional.
Group 2, composed of two isolated stations, appeared to be
linked only because they were intermediate in almost all
parameters. However, both had the highest salinity and second
highest DO, suggesting ocean influence. Group 3 stations
appeared to be more heavily impacted in terms of species
(TWT, Figure 27), than Group 1, even though Group 1 had the
lowest phytoplankton measures.
October 1978.
Prior to the final benthic sampling for TITP, the plant
upset had apparently been brought under control, in part by
added aeration, and dissolved oxygen had risen somewhat. In
the October station groups (Figures 29, 30) station A1 was
separated outside the harbor, and there were almost concentric
arcs out from TITP. Most interesting is Group 5 (Al4), which
lies at about the center of the large harbor gyre. Vorticity
apparently led to deposition, for the TWT (Figure 31) shows
this formerly rich benthic area to have been severely depleted,
perhaps by the rapid changes in nutrients and probably in
controlling substances such as chlorination. However, A14
was higher in chlorophyll a (Tables 15, 16) than the other
-------
IVB 7
413
stations, even though all are low for the fall; A14 was also
the highest in mean temperature. The physical variables,
however, did not have significant coefficients, whereas the
phytoplankton measures did.
Group 3, the outfall area, had the lowest weighted means
for DO, pH, productivity and assimilation ratio. Yet the two
stations had a significant increase in colonization, as shown
on the HIT. This perhaps emphasizes that variability, whether
natural or man-made, has a stimulatory effect on estuarine
species when toxicity is not a problem. There are no indica-
tions that normal TITP effluent is toxic (Section V).
FOr the first time since June 1977 the three phytoplankton
parameters assumed the major importance, even though the levels
were relatively low as compared with pre-secondary treatment
years. Groups 1 and 2 were intermediate in most parameters.
However, separations were clear in that Group 2 had higher
phytoplankton measurements than group 1.
LITERATURE CITED: See Section VIA.
METHODS SECTION, DISCRIMINANT ANALYSIS: See Section VIB.
-------
414
IVB 8
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Harbors Environmental Projects
University of Southern California
Figure i. Benthic Station Groups, December 1976.
Group i - A2, A12, A9	Group 3 - ai
Group 2 - A3, as, ah	Group 4 - A4, A7
-------
IVB 9
415
Figure 2.
TERMINAL ISLAND TREATMENT PLANT B ENTHICS ** DECEMBER, 1976
DISTANCE
200
...4-
160
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120
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80
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GROUPS
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76 1 202/A01
751202/A04
76 1 202/A07
3
4
-------
IVB 10
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AXIS 2
©
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AXIS 3
s
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Figure 4. Station Groups and Axes with Vectors, December 1976.
-------
Table i.
WEIGHTED CROUP MEANS
TERMINAL ISLAND TREATMENT PLANT OENTHICS +* DECEMBER. 1976
GROUPS
1
1.	DEPTH
2.	TEMPERATURE
3.	SALINITY
4.	OXYGEN
5.	PH
6.	PRODUCTIVITY
7.	CHLOROPHYLL A
0. ASSIMILATION RATIO
0.2665
17.2286
32.9708
7.	1065
8.	1208
2.0659
1.	1770
1.6573
7 . 66 04
17.2003
32.9635
6.9721
8.1207
1.9471
1.1901
1.5675
7
.8601
6.8531
17
.2310
17.1613
32
.9659
32.8047
7
• C 2 89
6.5838
e
.1116
8.1324
i
.6605
I .7224
l
. 1363
1.4886
i
.3668
1.3207
** ONE-WAY ANOVA FOR EACH VARIABLE ~ DF =	3, 32
VARIABLE	F
1.	DEPTH	0.10
2.	TEMPERATURE	0.01
3.	SAL IN1TY	0.14
4.	OXYGEN	0.08
5.	PH	0.01
6.	PRODUCTIVITY	0.05
7.	CHLOROPHYLL A	0.16
e.	ASSIM1LATIQN RATIO	0.07
-------
IVB 13
419
Table 2.
Terminal Island Tre\fment Plant Benthics, December 1976.
*» LATENT ROOTS At 10 SIGNIFICANCE TEST FOR EACH AXIS	**
AXIS ROOT X CUM * CHI SQUARED	OF
1	6.714E-02 66.9 66.9 I.88	10
2	3.0096—02 30.0 96.9 C.86	0
3	3.15 8E— 03 3. 1 100.0 0.09	6
VALUE)) *~
1976
AXES	12	3
1.
DEPTH
3.1
0.1
4Q.9
2.
TEMPERS T JRE
o
•
o
3.4
24.5
3.
SAL INI""1
46 .4
2. 1
0.6
4 .
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1 3 .4
2.0
12.3
5.
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4 .1
2.1
0.4
6.
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I .7
3Q. 1
12.0
7.
CHLOROPHYLL A
5.1
8.3
2.4
3 •
ASSIMLATICN RATIO
16 .0
43.9
IS
•
<0
COEFFICIENTS OF SEPARATE DETERMINATION (X lOO/SUM(AeS
TERMINAL ISLAND TREATMENT PLANT BENTHICS ** OECEMBER«
-------
420
IVB 14
WILMINGTON
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vroup
B1«
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Harbors Environmental Projects
University of Southern California
Figure 5. Benthic Station Groups, March 1977.
Group 1 - A3, as, ah Group 3 - A2, A12, A4 , A9
Group 2 - ai	Group 4 - A7
-------
IVB 15
421
Figure 6.
TERMINAL ISLAND TB2ATMENT PLANT BENTHICS ** MAfiCH, 1977
DISTANCE
group:
200
..4
160
"4
120
- -4
80
-4
40
0
4
77 0309/A03
770309/A11 1
77 0309/A 08
77 0309/A 01 2
770309/A02
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77 0309/A 04
770309/A09
770309/A07 4
3
-------
422
IVB 16
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IVE 17
423
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Figure b.
Station Groups with Axes and
(DASHED LINE INDICATES VECTOR
Vectors, March 1977.
IS NOT CLEAR-CUT)
-------
T ABLE 3•
WEIGHTED GROUP MEANS
TERMINAL ISLANO TREATMENT PLANT 8ENTMICS ** MARCH, 1977
ro

GROUPS
1
?.
3
4
1.
OEPTH
9.922B
11.5234
10.1454
4.7286
2.
TEMPERATURE
13.3557
13.2030
13.3569
13.7475
3.
SAL INITY
3 1.0100
31.6091
31.8076
31.7142
4.
OXYGEN
8.0252
7.7618
7.9092
6.44 11
5.
PH
6. 1 535
6.1439
a.1440
7.9077
6.
PRODUCTIVITY
13.6801
11.9738
12.0499
6.3191
7.
CHLOROPHYLL A
5.1573
4.6069
4.9025
8.2406
a.
ASSIMILATION RATIO
2.3976
2.2463
2.3498
C.8078
** ONE-WAY ANOVA FOR EACH VARIABLE * OF •= 3. 32

VARIAQLE
F
1 .
OEPTH
0.47
2.
TEMPERATURE
0.44
3.
SALINITY
1 .29
4 .
OXYGEN
1 .04
5.
PH
2.21
6.
PRODUCT I V1TY
0.16
7.
CHLOROPHYLL A
0.55
B.
ASSIMILATION RATIO
0 .69
-------
IVB 19
425
Table 4. Terminal Island Treatment Plant Benthics, March 197?
** LATENT ROOTS ANO SIGNIFICANCE TEST FOR EACH AXIS **
AXIS ROOT	X	CUM %	CHI SQUARED	OF
1	5.211 E + 00	99.0	99.0	52.96	10
2	4.002E-02	0.8	99.8	1.14	8
3	1.13 IE—02	0.2	100.0	0.33	6
COEFFICIENTS OF SEPARATE DETERMINATION (X 100/SUM{ABS VALUE)) *»
TERMINAL ISLAND TREATMENT PLANT BENTHICS ** MARCH, 1977
AXES
1
1.
DEPTH
1 .5
16.2
I 0.5
2.
TEMPERATURE
2 .8
36.8
1 1 .6
3.
SAL INITY
12 .9
0. 1
4.2
4 .
OXYGEN
2 .2
3.4
5.3
5.
PH
35 .9
0.5
0 .5
6.
PROOUCTIVITY
I 0 .5
19.1
31 .7
7 .
CHLOROPHYLL A
29 .3
10.5
18.4
a.
ASSIMILATION RATIO
o
.
in
13.5
17.8
-------
426
IVB 20
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SAN
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B10«
Group 1 *'2
A10»fe.
B1»
A9«
Group
AO*
Harbors Environmental Projects
University of Southern California
Figure 9.	Benthic Station Groups, June 1977.
Group 1	- A2, A3, ah, A12 Group 4 - ai, as
Group 2	- A9	Group 5 - A7
Group 3	- A4
-------
IVB 21
427
Figure 10.
TEEMINAL ISLAND TREATMENT PLANT 3ENTHECS ** JUNE, 1977
DISTANCE
200
— 4 -
160
.-4-
120
-.4-
80
.4.
40
- J-
GROUP
0
4
770609/A02
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77 06 09/A03
77060 9/A11
770609/A09
770609/A0£t
77 0609/A01
77 0609/A08
770609/A07
2
3
-------
428
IVB 22
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IVB 23
429
©
l
AXIS 1
FIGURE
12. Station Groups and Axes with Vectors, June
1977 .
-------
Table 5.
WEIGHTED GROUP MEANS
TEH MINAL ISLAND TREATMENT PLANT 6ENTHICS *~ JUNE,
GROUPS
1
1.	DEPTH	9.2711	9.0635
2.	TEMPERATURE	16.7837	16.3349
3.	SALINITY	33.8795	33.8777
4.	OXYGEN	7.0976	7.Q989
5.	PH	7.9656	7.9S64
6.	PRODUCTIVITY	12.7388	12.3125
7.	CHLOROPHYLL A	1.8734	1.8833
8.	ASSIMILATION RATIO	7.4374	7.0730
~» ONE-WAY ANOVA FCR EACH VARIABLE * OF =	4,

VARI ABLE
F
1.
DEPTH
0.08
2.
TEMPERATURE
0.12
3.
SAL IN ITY
0.15
4 .
OXYGEN
0.0 1
5 .
PH
C . 1 8
6 .
PRODUCT IVITY
0.02
7.
CHLOROPHYLL A
0.15
8.
ASSIMILATION RATIO
0.07
1977
4*
U>
O
8.8453
17.2284
33.86 13
7 . 1388
7.94 57
12.4927
1 .9604
6.8968
9.4506
17.2248
33.8710
7.I 133
7.9615
12.4914
1.8853
7.1394
7.3783
18.1485
33.8358
7.0629
7.8984
13.2675
2.3265
5.9545
s
K>
40
-------
IVB 25
431
Table 6. Terminal Island Treatment Plant Benthics, June 1977.
** latent roots and significance test fcr each axis »*
AXIS ROOT	X	CUM X	CHI SOUARED	OF
1	3.025E-01	80. 7	80.7	<3.91	11
2	3.821E-02	10.2	90.9	1.41	9
3	2.836E-02	7.6	96.4	1.05	7
4	5.902E-03	1.6	100.0	0.22	5
COEFFICIENTS OF SEPARATE DETERMINATION (X 10C/SUM(A8S VALUE)) *+
TERMINAL ISLAND TREATMENT PLANT 8ENTHICS ** JUNE. 1977
AXES	12	3	4
1.
OEPTH
2 1 .6
20.5
4 1.9
2.4
2.
TEMPERATURE
1 1 .1
50.3
2.0
0.4
3.
SAL INITY
9 .0
8.2
10.1
1 .S
4.
OXYGEN
1 .3
I 1 . 0
8.3
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7.8
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6.
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0 .3
0. 1
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35.3
7.
chlorophyll a
43 .7
6.4
25.7
28.6
a.
ASSIMILATION RATIO
5 .3
1 .3
0 .6
26.2
-------
432
IVB 26
01*
B3*
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A7
SAN
PEDRO'
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A16»
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A2»
A10*
A13
B1»
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¦roup 4
A1*
Harbors Environmental Projects
University of Southern California
Figure 13. Benthic Station Groups, September 1977.
Group 1 - A2, A9, ai3	Group 4	- ai, Ai4
Group 2 - A3, A12, A15, A16, A17 Group 5	- A7
Group 3 - A4, as
-------
IVB 27
433
Figure 14.
TERMINAL ISLAND TREATMENT PLANT BENTHICS ~* SEPTEMBER# 1977
DISTANCE
200
-.4-
160
120
..J.
80
-I-
40
-l-
0
GROUPS
77 0914/A02
77 0914/A13
77 0914/A09
77 091U/A12
77 0914/A 16
77 091VA 15
77 0914/A03
77 09 1 U/A 17
77091 4/A0U
77 091 4/A 03
77 0914/AQ1
77 0914/A 1t|
77 091 4/A 07
4
5
-------
434
IVB 28
TittniNAl 1SLAIU TrffiATftgiT PLktt dtlTBICS ~ • SXPTEB3EB, 1977
Figure :o.
>	.75 to	i
>	.SC TO	.75
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IVB 29
435
AXIS 2
©
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a p
Ai

D
-~ pH
Sal.
— Depth
Prod.
+ Chi A
AXIS 1
Figure 16. Station Groups and Axes with Vectors,
September 1977
-------
Table 7.
.t*
u>
-------
IVB 31
437
Table 8. Terminal Island Treatment Plant Benthics, September 1977
** LATENT ROOTS AND SI C-N[F icance test for each axis **
AXIS ROOT	X	CUM X	CHI SQUARED	OF
1	1.1046+00	98.6	98.8	42.76	II
2	1*01 IE—02	0.9	99.7	0.58	9
3	2.9<53fc-03	0.3	99.9	0.17	7
4	6.221 E— 0 4	C.l	100. 0	0.04	5
COEFFICIENTS OF SEPARATE DETERMINATION (X 100/SUM(ABS VALUE)) **
TERMINAL ISLANU TREATMENT PLANT BENTHICS ** SEPTEMBER, 1977

AXES
1
2
3
4
1.
DEPTH
31 .7
I .3
13.6
46.4
2 .
TEMPERATURE
6.2
11.6
43.2
11.7
3.
SAL INITY
31.7
i .2
0.9
£.6
4 .
OXYGEN
6 .7
17.0
2.6
2.8
5.
PH
15.0
21.0
14.6
24.4
6.
PRODUCTIVITY
3 .3
5.7
12.2
0.3
7 .
CHLOROPHYLL A
4 .5
a. 3
10.7
6.5
H .
ASSIMILATION RATIO
O
•
o
34.0
2.2
1.4
-------
438
IVB 32
-B6-
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#1
roup 3
Harbors Environmental Projects
University of Southern California
Figure 17. Benthic Station Groups, January 1978
Group i - as, A9, aio	Group 3 - ai
Group 2 - A2,A3, Ai2, Ai3, ai« Group w - A4, ah, ais
A 1 6 , A I 7 , B8, B9	GROUP 5 - A7
-------
IVB 33
439
Figure 18.
TSBMINAL ISLAND TREATMENT BENTHICS ** JANUABY, 1978
DISTANCE	GROUPS
200 160 120 80 HO 0
- « 4	4	4	4	4	4
78 0106/A 08
780106/A10 1
780106/A09
780106/A12
780106/AU
78 0106/A16
78 0106/B08
78 0106/A17 2
78 01 06/A 02
780106/A13
780106/A03
780 106/B09
780106/A 01 3
780106/A04
78 0106/A15 4
780106/A11
78 0106/A07 5
-------
440
IVB 34
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-------
IVB 35
441
AXIS 2
S>
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©
©
©
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Sal.
. DO
Pvod.
©
AXIS 2
©
©
AXIS 1
©
@ Pd
Frod
©
AXIS 3
Figure 20. Station Groups and Axes with Vectors, January 19
-------
Table 9.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT BENTHICS ** JANUARY, 1978
ro
GROUPS
1

2

3
4
5
1 .
DEPTH
10.6365

10.3337
1 0
.3797
10.1784
7.8126
2.
TEMPERATURE
16.0046

16.0003
15
.9950
16.0005
15.8866
3.
salinity heavy rains l%t
27.5755

27.5688
27
.5725
27.5552
27.3806
4 .
OXYGEN
7.3456

7.2762
7
.261 1
7.1817
5.9463
5.
PH
8. 1 027

8.0690
7
.9601
8.0667
8.0068
6.
PRODUCT IV ITY
2. 1401

2.2260
2
.2930
2.2682
2.1718
7.
CHLOROPHYLL A
1.2220

1 .2368
1
.2438
1 .2296
1.2114
8.
ASSIMILATION RATIO
1.7256

1 .7761
1
.8 15S
1 .8212
1-7752
ONE
-WAY ANOVA FOR EACH VAR
I ABLE *
DF =
4 .
80




VARIABLE
F






1 .
DEPTH
0
.39





2.
TEMPERATURE
0
.77





3.
SALINITY
0
.74





4 .
OXYGEN
1
.37





5 .
PH
0
.09





6.
PRODUCTIVITY
0
.03





7 .
CHLOROPHYLL A
0
.01





8 .
ASSIMILATION RATIO
0
.03





H
<
W
U)
G\
-------
IVB 37
443
Table 10. Terminal Island Treatment Plant Benthics, January 1978
** LATENT ROOTS AND SIGNIFICANCE TEST FDR EACH AXIS **
AXIS ROOT	X	CUM *	CHI SQUARED	OF
1	2.873E-C1	92.8	92.8	19.57	II
2	1.366E-02	4.4	97.3	1.05	9
3	7.867E-03	2.5	99.8	0.61	7
4	6.1725-04	0.2	100.0	0.05	5
COEFFICIENTS OF SEPARATE OETERMINATION (X 100/SUM(A8S VALUE)) ~*
TERMINAL ISLAND TREATMENT 8EMTHICS *< JANUARY, 1976
AXES IN COLUMNS	12	3	4
1.
DEPTH
•
CD
0.8
e.9
11.1
2.
TFMPERATURE
26 .6
1 .3
19.6
0.3
3 .
SAL INI TY
1 0 .5
0.3
26.0
0.6
4 .
OXYGEN
56 .5
0.3
6.2
C . 0
5 .
PH
0 .9
73 . 9
0.3
0.5
6 .
PRODUCT I V ITY
0 .2
8.3
18.5
37.2
7 .
CHLOROPHYLL A
0 .5
1 .2
0.0
29.5
0.
ASSIMILATION RATIO
0 .1
13.9
20.5
20.7
-------
444
IVB 38
WILMINGTON
LONG BEACH
',Coio^-.

SAN ,'A n*5'
•' fSi'\
PEDRO
AIO*

AO*
Harbors Environmental Projects
University of Southern California
Figure 21. Benthic Station Groups, April 1978
Group 1 - A3, aio, A14, ais, ai6, ai?, b9
Group 2 - ai, A2, as, A9, aii, A12, Ai3, bs
Group 3 - A4, A7
-------
IVB 39
445
Figure 22.
TERMINAL ISLAND Tfi&ATMENT PLANT DSNTUICS ** APRIL, 1978
DISTANCE
200
..4.
160
..4.
120
..4.
60
.4-
40
.4.
0
4
78 0410/A 14
780410/A17
78 0410/B09
78 0410/A10
780410/A 15
780410/A03
78 0410/A16
GRO
78 04 10/A13
78 0410/B08
780410/A02
78 0410/A08
78 0410/A09
780410/A 12
7804 10/A11
78 0410/A 01
78 0410/A04
780410/A07
-------
446
IVB 40
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-------
IVB 41
447
A XI S 2
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Figure 24.
Station Groups and Axes with Vectors, April
1978
-------
TABLE 11.
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT PLANT BENTHICS ** APRIL. 1978
.u
00


1
2
3
1.
DEPTH
10.6248
10.9074
10.0003
2.
TEMPERATURE
16.1917
16.1696
16.2871
3.
SALINITY
35.1184
35.1922
35.1727
4 *
OXYGEN
9.2477
9.3961
9.1449
5.
PH
7.7294
7.7211
7.7239
6.
%TRANSMITTANCE
64.7771
65.1413
63.3858
7.
PROOUCTIVITY
3.3230
3.3004
3.5117
a.
CHLOROPHYLL A
I.5860
1.5836
1.6050
9.
ASSIMILATION RATIO
2.0889
2.0893
2.2207
~~ ONE-WAY ANOVA FOR EACH VARIABLE * OF =	2. 48

VARI ABLE
F
1 .
DEPTH
0.04
2.
TEMPERATURE
0.02
3.
SALINITY
0.01
4.
OXYGEN
C .03
5.
PH
0 .0 I
6.
fcTRANSMITTANCE
0.02
7.
PRODUCTIVITY
0.01
8.
CHLOROPHYLL A
0.00
9.
ASSIMILATION RATIO
0.01
-------
table 12. Terminal Island treatment plant Benthics, april 1978
#* latent roots and significance test for each axis ~*
AXIS	ROUT	X	CUM X CHI SQUARED	DF
1	9.223E-03	73.9	73.9	0.40	10
2	3.257E—03	26.1	100.0	0.14	8
COEFFICIENTS OF SEPARATE DETERMINATION IX 100/SUM(ABS VALUE!) ** (AXES IN COLUMNS)
TERMINAL ISLAND TREATMENT PLANT BENTHICS ** APRIL, 1978
1	2
1.
DEPTH
19.7
3.7
2.
TEMPERATURE
20.7
0.4
3.
SAL INITY
0.9
25.5
4.
OXYGEN
7.4
25.6
5.
PM
1 .7
36.4
6.
XTRANSMITTANCE
15.0
0.8
7.
PRODUCTIVITY
12.3
3.2
6.
CHLOROPHYLL A
3.4
0.0
9.
ASSIMILATION RATIO
Id.9
4.3
.U
•to
U3
-------
450
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Harbors Environmental Projects
University of Southern California
Figure 25. Benthic Station Groups, July 197a.
Group 1 - A2, A3, A9, aio, ah, Group 4 - A4
A12, a 15, Aie, a 17	Group 5 - A?
Group 2 - ai, as
Group 3 - A13, Ai4
-------
IVB 45
451
Figure 26.
TERMINAL ISLAND TfiEATHENT PLANT BENTHICS ~* JUL*, 1978
DISTANCE
200
..J.
160
120
80
.*•
10
0
4
GROUPS
780710/A09
78 07 10/A 10
780710/B08
780710/B09
780710/A02
780710/A12
780710/A17
780710/A03
780710/A16
780710/A11
780710/A15
78 0710/A01
78 0710/A08
78 0710/A13
780710/A14
780710/AOI4 4
780710/A07 5
-------
452
IVB 46
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-------
IVB 4 7
453
AXIS 2
©
¦AR
Temp,
DO
pH
©
©
3
Devth
AXIS 1
®
Figure 28.
Station Groups and Axes with Vectors,
July 1978.
-------
Table 13.
•t*
Ul
WEIGHTED GROUP MEANS
TERMINAL ISLAND TREATMENT PLANT 8ENTHICS ** JULY. 1978
GROUPS	12	3	4	5
1.	DEPTH	10.2844	10.0290	10.6857	9.2064	8.3176
2.	TEMPERATURE	12.9937	13.0890	12.8468	13.2904	13.6241
3.	SALINITY	31.4894	31.4950	31.4920	31.4753	31.4324
4.	OXYGEN	5.0035	5.1351	4.9645	5.1253	5*6984
5.	PH	8.0487	8.0536	8.0416	8.0703	8.1102
6.	PRODUCTIVITY	4.1364	4.1962	4.1943	4.7861	4.2700
7.	CHLOROPHYLL A	4.6817	4.6391	4.6888	5.2902	5.2324
8.	ASSIMILATION RATIO	0.9009	0.8810	0.8358	0.9046	0.8031
H
<
(D
00
»* ONE-WAY ANOVA FOR EACH VARIABLE * OF - 4, 80

VARIABLE
f
1.
DEPTH
0.27
2.
TEMPERATURE
0.22
2.
SAL INITY
o
•
CO
4 .
OXYGEN
0.2 1
£ .
PH
0.36
6 .
PRODUCTIVITY
C .07
7.
CHLOROPHYLL A
0.15
8.
ASSIMILATION RATIO
0.05
-------
IVB 49
455
Table 14. Terminal Island Treatment Plant Benthics,	July 1978
»* latent roots and significance test for each axis	**
AXIS	ROOT	%	CUM % CHI SQUARED	DF
1	9.329E-C2	75.3	75.3	6.91	41
2	1 .860E-02	15.0	90.4	1.43	9
3	9.237E-03	7.5	97.0	0.71	7
4	2.699E-03	2.2	100.0	C.21	5
COEFFICIENTS OF SEPARATE DETERMINATION (X lC0/SUM(AeS VALUE)) **
TERMINAL ISLAND TREATMENT PLANT BENTHICS ~* JUL*. 1978
AXES IN COLUMNS	12	3	4
1.
DEPTH
24.1
1 .7
4 .5
4.7
2.
TEMPERATURE
1 5 .0
19.7
8. 1
1.0
3.
SAL INI TV
20 .5
• 5.5
0.4
24.6
4.
OXYGEN
23 .1
23. 8
3.2
49.0
5 .
PH
12.5
0.8
4.3
2.. 0
6.
FRCJDUCTI VI TY
2 .6
4.8
40.4
1 .0
7.
CHLOROPHYLL A
C .9
25.6
0.3
17.0
8.
ASSIMILATION RATIO
1 .1
18.0
38.a
0.8
-------
456
IVB 50
WILMINGTON
LONG BEACH
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A7
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PEDRO;
A12i
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B1«
Harbors Environmental Projects
University of Southern California
Figure 29. Benthic Station Groups, October 1978.
Group 1 - A3, as, ah, A12,	Group 3 - A4, a?
a 15, a 16, B8	Group 4 - ai
GROUP 2 - A 2, A9, A10, A13, A17, B9	GROUP 5 - A14
-------
IVB 51
457
Figure 30
TERMINAL ISLAND TREATMENT PLANT BENTHICS ** OCTOBER, 1978
DISTANCE	GROUP
200 160 120 80 40 0
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-------
458
IVB 52
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-------
IVB 53
459
AXIS 2
ft
Depth
Sal.
Temp,
Chi A
© ©
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i)	i
AXIS 1
Figure 32. Station Groups and Axes with Vectors,
October 1978.
-------
Table is.
C- H T 6 0 GROUP MEANS





!M INAL ISLAND TREATMENT
PLANT BENTHICS
*~ OCTOBER,
1978


GROUPS
I
2
3
4
5
1 .
DEPTH
1 1.5537
1 I .704 0
10.9726
13.4696
I 1.2970
2 .
TEMPERATURE
1 7.2055
16.9490
17.4641
16.9706
18.0490
3.
SAL INITY
3 1.2979
31. 3 04-9
31.2935
31.3383
31.2724
4.
OXYGEN
6. 7870
6.7799
6.8307
6.7787
6.7227
5 .
PH
8. 1209
6. 1 107
8. 1377
a.103 I
8.1 175
6.
PRODUCT I VITY
6.6967
8.8114
8.8250
7.9410
7.6540
7 .
chlorophyll A
3.4523
3.4673
3.4304
3.2 2 73
4.2247
e.
ASSIMILATION RATIO
2.6393
2.6685
2.6746
2.5382
2.0406
CNE
-WAY ANOVA FOR EACH
VARIABLE ~ DF =
CO




VARI ABLE
F




1 .
DEPTH
C * 2 0




2 .
TEMPERATURE
o
•
o




2.
SAL INITY
0.17




4 .
OXYGEN
0.00




E.
PH
o
*
o




6 .
PRODUCTIVITY
<0
o
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. CHLOROPHYLL A
0.47




a.
ASSIMILATION RATIO
0.11




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-------
IVB 55
461
Table 16.
** LATENT ROOT S ANC SIGNIFICANCE TEST FOR EACH AXIS **
AXIS ROOT	%	CUM X	CHI SQUARED	OF
1	1.373E-01	70.5	70.5	9.72	1 1
2	4.915E-02	25.2	<55.8	3.62	9
3	7.617E-03	3.9	9 <3.7	0 .57	7
4	6.219E-C4	0.3	100.0	0.05	5
COEFFICIENTS OF SFPARATE DETER Ml NAT ION (X 100/SUH(AqS VALUE)) **
TERMINAL ISLANO TREATMENT PLANT BENTHICS ** OCTOBER, 1978
AXES IN COLUMNS
1
2
3
4
1 .
DEPTH
0 .4
44.3
2 . 0
6.9
2.
TEMPERATURE
I . 1
4 . 7
a. 5
3.3
3.
SALINITY
0 .9
4.4
3.6
3 1.9
4 .
OXYGEN
0 .0
0.2
a .2
7.0
E .
PH
0 .2
2.0
31 .6
22. a
6.
PRODUCTIVITY
1 S .9
17.4
35.2
13.8
7.
CHLOROPHYLL A
4 1 .4
17.5
t .5
3.5
e.
ASSIMI LA TICN RATIO
36.9
9.4
9.4
10.9
-------
Intentionally Blank Page
-------
VA
463
PHYTOPLANKTON GROWTH AND STIMULATION IN THE
TERMINAL ISLAND TREATMENT PLANT SECONDARY WASTE PLUME
INTRODUCTION
The Terminal Island Treatment Plant {TITP) releases 10-18
million gallons of secondary treated sewage effluent into the
Los Angeles Harbor daily. This series of algal bioassays was
designed to assess the impact of TITP effluent on phytoplankton
growth. The culture data presented here gives a clear indica-
tion of the growth response of the harbor phytoplankton com-
munity to effluent levels actually found in the harbor.
METHODS
Diatoms and dinoflagellates dominate the harbor phytoplank-
ton community. Therefore, a mixed culture of harbor diatoms
(Skeletor.ema, Nitzsehia and naviculoid species) and a monoculture
of the harbor dinoflagellate Sovi-ppsiella troahoidea were used
in the initial experiments of the bioassay series. The growth
response of the mixed diatom culture to any given test solution
was highly variable. Possibly, this was due to competitive in-
teractions between diatom species in the spatially restricted
environment of the laboratory culture. However, some differen-
tial growth response was observed. Sarippsiella troahoidea, an
extremely slow-growing species, did show a growth response in
one 7-day bioassay in February. It did not, however, show a
differential growth response to any of the experiments during
an 8-day June bioassay period. Therefore, use of this species
was discontinued.
Dunaliella tertiolesta was selected for use in each monthly
bioassay. This microflagellate is present in the harbor and it
has been widely used in the past as a bioassay organism. It
grows rapidly and was found to be a sensitive indicator of growth
conditions.
Experimental Design
Dilution for growth tests were chosen to encompass the range
of effluent concentrations found in the harbor in each sampling
period. The percentages of effluent concentrations chosen con-
form to a logarithmic progression, with replicates prepared for
each test. Six concentrations of TITP effluent (10%, 5.6%, 3.2%,
1.0%, 0.56%, and 0.1%) were tested in each period along with wa-
ter from the harbor taken from over the TITP outfall and from
three other stations in the Los Angeles Harbor, A7, A3, and A2,
which are approximately 550m, 1525m, and 1875m, respectively,
from where the effluent surfaces from the plant in a turbulent
circle of water known as the "boil." Three additional samples
Preceding page blank
-------
464
VA 2
served as controls. In addition, a dilution water sample was
tested to determine minimal growth in the absence of effluent
stimulation or inhibition, and a sample containing an enriched
algal medium was used to assay maximal growth under optimum
nutrient conditions in order to check on the health of the orig-
inal inoculum culture. An Instant Ocean artificial sea water
was also tested to determine minimal growth in the absence of all
extraneous nutrients. However, growth in Instant Ocean was high-
ly variable from one bioassay to another, suggesting variable
nutrient content.
Preparation of Test Solutions
Each effluent concentration was prepared using TITP efflu-
ent obtained from inside the plant on the first day of each bio-
assay test series. The effluent was corrected for low salinity
with Instant Ocean sea salts.
In the February bioassay, the effluent was diluted using
ultraviolet-sterilized and filtered sea water from the Harbors
Environmental Laboratory at Wilmington. A1 subsequent bioassays
utilized aged dilution water collected in a single batch from
midchannel in the San Pedro Bight. Each assay solution was
filtered through a GF/C glass filter. A 10 0 ml volume of this
filtered solution was then transferred into each of three 250 ml
Nalgene Ehrlenmeyer flasks.
Bioassay Procedures
The phytoplankton inoculum was grown in axenic culture me-
dium for one week prior to the bioassay. After cell densities
were determined, 10 ml of the healthy algal culture was inocu-
lated into each 100 ml of test solution prepared as described
above.
The culture flasks were maintained in a seawater table under
constant light (approximately 40 microeinsteins/meter^/sec.) and
temperature (18.5C±5) for the 5-day bioassay period. Each flask
was shaken, and its position in the table rotated daily. At 2-
day intervals subsamples were removed from each flask and pre-
served in Lugol's solution. Cell densities were subsequently
determined with a Coulter Counter.
This bioassay procedure was performed five times at bimonth-
ly intervals in February, April, June, August, and October 1978.
RESULTS
Final Cell Yields
Representative growth curves for selected dilutions for
April are given in Figure 1. Final cell concentrations for all
-------
VA3
465
tests of Dunaiiella tertioleeta in the February bioassay are
given in Figure 2. The laboratory seawater supply was used for
dilution in the bioassay series. It was then recognized that
this water alone provided nutrient enrichment. However, signif-
icantly higher cell yields were found in effluent concentrations
of 1.0% or greater. The 3.2%, 5.6% and 10% dilutions were not
significantly different from each other. A high yield was also
produced in filtered water from station A7, located nearest to
the TITP outfall boil (see Figure 3). The yield at this station
was comparable to that found with the 1.0% TITP effluent. The
growth response in the outfall boil tests was highly variable.
Possibly, this was due to high numbers of bacteria that compete
with the diatoms and dinoflagellates in this nutrient-rich medi-
um. Scrippsiella troahoidea (Figure 4) and the mixed diatom
culture (Figure 5) also showed increased yield at concentrations
of 1.0% TITP effluent or greater. Cell yields of S. troahoidea
were also increased in water from the four harbor stations
(A2, A3, A7, and the boil itself). The highest cell numbers oc-
curred in water from A7, the closest of the three stations to
the outfall boil. For the mixed-diatom cultures, the yield in
the outfall boil sample was increased, but was not significantly
different from 1.0% dilutions of TITP waste in some instances and
up to 10% in other tests.
The final cell yields of four subsequent bimonthly bioassays
with Dunaiiella tertiolsota are given in Figures 6 through 9.
The composition of the TITP effluent is complex, with variations
in nutrients and possibly unidentified inhibitors or stimulators
present in varying amounts. However, the growth response ob-
served in this series of bioassays followed a consistent pattern.
Relative to the dilution water control, cell yields were
significantly increased by the addition of low concentrations of
TITP effluent (0.56-3.2%). In the April, June, and August 1978
bioassays, final cell yields increased with increasing TITP con-
centration up to and including 10%. However, in the October bio-
assay final cell yield in the 10% dilution was not significantly
different from the 5.6% dilution. The final cell yields in the
outfall boil water were usually comparable to those from the 10%
TITP solution, while average yields in the harbor station water
samples usually decreased with distance from the boil. In the
June bioassay, however, cell yield in the boil water sample was
below that of the experimental dilution water control, and aver-
age cell yields in the other harbor station treatments increased
with distance from the boil. These data suggest two possibili-
ties: 1) The presence of an unidentified inhibitor in the harbor
water diluting the effluent (if this is true, this inhibitor must
have been in higher concentrations near the boil than near the
outer harbor stations A3 and A2); or 2) The formation of an in-
hibitor resulting from an interaction between components of the
effluent and substances in the waters of the harbor. It is not
possible to distinguish definitely between these two alternatives
on the basis of the information available. However, the TITP
plant malfunctioned during the period in question, allowing raw
-------
466
VA 4
wastes to escape. Chlorination was also heavy in that period.
Specific Growth Rates
Final cell yields are useful as a means of comparison among
the tests. In this series of bioassays, high initial cell con-
centrations were used to insure detection of any stimulatory
or inhibitory effect. Therefore, the final cell densities were
much higher than those that would normally be found in the har-
bor. Initial cell concentrations also varied somewhat from
one bimonthly bioassay to another.
In order to make comparisons between bioassays and the
growth of the natural phytoplankton populations, specific
growth rates were calculated. Specific growth rate (r) is de-
fined as tne rate of population increase per day.
r = log e Nt
No
t
where Nt = cell density at day t
No = cell density at day 0
t = total time interval in days
Average specific growth rates for all preparations in five
bioassays are given in Table 1.
The February bioassay differed from the four subsequent
bioassays in that the initial phytoplankton concentration was
much lower and the bioassay ran for seven days rather than eight.
Each of these factors would contribute to higher r values. Max-
imum short-term r values were observed during the June bioassay
in 10% TITP (r = .67) and in water from the boil (r = .68).
The bioassays were conducted under constant, fluorescent
illumination of 40 microeinsteins/m^/sec (0.0132 langleys/min.).
This is equivalent to approximately 2 percent of full midday
sunlight. Natural phytoplankton communities encounter comparable
light intensities within the upper five meters depth in the har-
bor (Kremer, personal communication). Sraayda (1973), however,
found that the diatom Skeletonema oostatum reached light satura-
tiontion of 0.15 langleys/min., a light intensity approximately
10 times greater. Specific growth rates also vary with tempera-
ture. Under optimum light conditions in culture D. tertioleata
reached a maximum specific growth rate of 1.2 3 at 18°C (Eppley,
1972). This value is approximately double the maximum short-
term growth rates in our experiments. Therefore, it is prob-
able that light limited the maximum specific growth rates obtained
here, and that r values would have been greater if light levels
were increased.
-------
VA 5
467
Natural phytoplankton from oligotrophia waters off southern
California showed r values of 0.17-0.28 at 20°C in simulated
in situ conditions. The maximum r expected for optimum conditions
was 1.5. In the nutrient-rich waters of the Peru Current, r
averaged 0.46 at 17-20°C, about half of the r expected under
optimum light conditions (Eppley, 1972). Even in those relative-
ly clear waters, light limitation appears to decrease r. Thus,
the light levels used in the bioassay appear to reflect actual
conditions in the harbor environment.
Nutrients
Marine phytoplankton require a variety of nutrients for
growth and reproduction. Phytoplankton growth is believed to
be limited by whichever factor is present in minimal quantity.
Nitrogen, phosphorus and silicon are potentially limiting to phy-
toplankton growth because they are not always present in excess.
Nitrogen and phosphorus are utilized in the synthesis of organic
materials at a ratio of 15N:1P. If the phytoplankton in the har-
bor assimilate nitrogen and phosphate in approximately this ratio,
phosphate is very unlikely to become limiting (see harbor nutri-
ent levels in Table 2}. Nitrogen has been shown to be the most
important nutrient that limits phytoplankton growth in marine
systems (Riley and Chester, 1971). According to Sverdrup, et at.
(1942), inorganic nitrogen is present in natural sea waters as
nitrate (.1-43 pg-at/1 NO3), nitrite (.01-3.5 pg-at/1 NO2) and
ammonia (.35-3.5 jug-at/1 NH3) . Nitrate is usually the most abun-
dant and stable source of nitrogen in oligotrophic (nutrient-
poor) waters. Ammonia is the energetically more efficient N form
and is preferentially absorbed when available (Harvey, 1955).
This form (NH3) may become the more important N source at times
(see Thomas, 1966). Secondary waste treatment usually elevates
nitrate and nitrite production, but may decrease ammonia (Dunstan
and Menzel, 1971).
Uptake of nitrate is believed to be suppressed when NH3-N
exceeds 1.0 pg-at/1 (Eppley et at., 1969). Until September 1978
NH3-N levels in the TITP effluent exceeded 150 pg-at/1. There-
fore NH3 probably provided the N utilized in phytoplankton
growth. The total inorganic nitrogen content of the TITP efflu-
ent during bioassay months is given in Table 2. These data indi-
cate that nitrogen levels in the 10% TITP bioassay treatment
ranged from 120 to 200 pg-at N/1. Nitrogen enrichment alone
could account for the increased growth rates in the TITP treat-
ment. Since September 1978 the total inorganic N levels have not
greatly changed. However, secondary treatment is now converting
most NH3-N to the NO3 form. Where nitrogen and phosphorus are
present in excess, other factors may become limiting, such as:
iron, manganese, copper, molybdenum, boron, vanadium, zinc, and,
for diatoms, silicon; all are required in small quantities and
may potentially become limiting.
-------
468
VA 6
CONCLUSIONS
Impact on the Harbor Phytoplankton Community
The introduction of TITP effluent into the harbor can be
viewed as local perturbation of the phytoplankton community.
In order to predict the spatial and temporal extent of the per-
turbation, we have applied the concept of critical length
(Kierstead and Slobodkin, 1953; Steele and Mullin, 1977). The
critical length of a unique patch of water is defined as the min-
imal size necessary for that patch to maintain itself despite the
dispersive process of mixing. The critical length is given by:
lc = vfJT
where K is the coefficient of horizontal eddy diffusion and
r is the specific growth rate characteristic of phy-
toplankton in the patch.
This model incorporates factors reflecting the unique chemical,
biological (r) and physical (K) characteristics of the patch.
The specific growth rates characteristic of phytoplankton
in the TITP plume have been empirically determined by bioassay.
When current speeds are known, diffusion coefficients for a
point source are given the following equation from Foxworthy
and Kneeling (1969):
where u = average current speed
= the mean variance of the waste concentration
distribution in a given coordinate direction
as a function of distance (x) along a plume
discharged from a point source.
Tidal circulation within the harbor is weak. The currents
in the area of the TITP plume are primarily wind-generated, av-
eraging .1-.2 knots or approximately 5-10 cm/sec (Robinson and
Porath, 1974; McAnally, 1975). Using Foxworthy and Kneeling
values for s2 and substituting these current speeds into the a-
bove equation, we have generated diffusion coefficients ranging
in magnitude from 2 x 10^ to 2 x 10-*. These values can then be
used in the critical length model.
Increased specific growth rates were detected at effluent
concentration of 1.0% or greater. Using 1% TITP to define the
limits of a patch, estimates of critical length have been com-
puted below.
-------
VA 7
469
Critical Length LC = (meters)
r(day~l) r(hr~l) where K - 2 x 10^ K = 2 x 103
1% max. r
1% mean r
.28
.55
.01
.02
444
314
1405
993
According to this model, when the 1 percent dilution patch ex-
ceeds the critical length, the effects of the effluent plume will
persist over significant scales of time (several days) and dis-
tance (several kilometers). If harbor nutrient levels are used
as an index of dilution, the 1 percent dilution level falls be-
tween station A7 and A3, 550 meters and 1525 meters from the
TIT? boil, respectively (see Table 2 and Figure 10). This infer-
ence is strengthened by the bioassay findings that phytoplankton
growth in water from station A7 was comparable to that of the
1	percent treatment (see Figures 8 and 9). Thus, the dimensions
of the 1 percent plume do at times exceed the critical lengths
generated by the model, and the effluent patch can be expected
to persist for several days, enough time to produce a local in-
crease in the phytoplankton crop. Through continued dispersive
losses, this locally persistent patch would contribute to ele-
vated phytoplankton densities in the harbor.
Bioassay tests using various cultures of phytoplankton were
conducted at bimonthly intervals during 1978 to determine the
effect of the waste waters on growth rates. Concentrations
of 0.1 to 10% waste water from Terminal Island Treatment Plant
were tested and surface waters from four stations extending from
the boil to the breakwater were sampled for comparison.
The general pattern found was one of increasing growth rate
with increasing concentrations of waste water. The 1 percent
concentration appeared to be the level above which the growth
rate increased most sharply. Goldman and Stanley (1974) found
that Dunaliella teriiclsota did not increase in biomass in cul-
ture at concentrations of more than 20 percent sewage.
Station A7, about 525 meters from the boil, showed growth
rates comparable to those found in the 1 percent solution. Us-
ing measured nutrient concentration as an index of dilution, the
2	percent level would lie between station A7 and station A3,
about 1525 meters from the boil. This suggests that in 1978 the
zone of enhanced phytoplankton productivity extends only to about
500 to 1500 meters from the boil.
LITERATURE CITED See Section VI
-------
470
VA 8
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-
1 1 1 1 1
l l I
i
i
Q
4 5 6
Days
8 9 10
-------
DlI. S.W.
FY 2
Bo I I
ctn A2
s tn A3
stn A7
In«t. 0.
0. t J<
0.56X
I .0 X
3.2X
5.6X
10X
H
Figure 2
PUNALIELLA C7th Pay - 20 Feb 78)
-ds-
-E^3-

> dp i
i—
H
0 100 208 300 400 500 600 700 800 900 1000 1100 1200
Thousand caI 1s par mI .
t» Th« d I ff«renc« Is slgnlfleont Cby onova, P< 0.001); Ronga S. moan ± 2 S.E. ar«s shown.
-------
472
VA 10
) lrws
4r SK
j>7 3 02
'dj
TITP = Treatment plant line
VS
SK
Way8 Street outfall
StarKist 4 outfall
1000 ft
Harbors Environmental Projeatg
University of Southern Cali
-------
Figure 4
SCRIPPSIELLA C7th Day - 20 F«b 78>
Bo I I
sin A2
tn A3
tn A7
0.01 X
0.56%
I . 2'A
3.2 %
5.6%
1080
2000
6000
7000
.6000
* Th« d i f*farcnc« I:
4000 5000
CttI Is par mI .
sign if Icant Cby anova, P< 0.001); Rang* & mtan + 2 S.E. ar« shown.
9000
-------
Dl i . S. W.
F/2
Bo I I
sia A2'
fit a A3
sta A7
Inct. 0,
0. J 'A'
0.S6X
i .ex
3.2 X
5. 6X"
10X
Ini tI a I
-|	1	1	1	r
V
0
Figure 5
MIXED DIATOMS C2nd Pay - 15 Fab 763
J	L.
H
H
-I
-1	I	I	I	I	I	1	1	I	L
8 10 12 14 16 18 20
Thoucand cat Is par mi ,
22
24
26
28
>F>
-J
.U
<
>
NJ
30
* Tha d i f* farenca Is significant Cby anova, P< 0.0013; Ranga & m«an + 2 S.E. are shown.
-------
Figure 6
PUNALIELLA C6lh Day - 25 April 785
Dl I . S.W.
t n A2
tn A3
ctn A7
0.1%
0.56%
. 0%
3.2%
5.6%
10%~
200
400 600 800 1000 1200 1400 1600
Thousand caI Is par ml .
1800 2000
2200
* Thtt d i f f«.-tnc9 Is significant Cby anova, P< 0.Q01 ); Rariga & maan + 2 S.E. are shown.
cn
-------
DlI. S.W. ~
F/2-
B o I I ""
stri A2 "
stn A3~
fitn A7"
Xnst. 0."
0. IX-
0.S6X-
I .0%"
3.2 X"
5.6%-
10X"
IPTTJ
1X3X1
Figure 7
DUNALIELLA C6lh Day - 27 Juna 78)
-1	1	1	1	*-1	1	1	
i rH i
rcpi
I—
I r4l t
I LI I I
-I	I	1	U
J	u
J	L

-J
Ol
<
>
150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 S50
Thousand c«I Is par ml .
* Th« dlff«r«nc» Is significant Cby onova, P< 0.001); Rang* & m«an + 2 S.E. ar* shown.
-------
Figure 8
Dl1. S.W. -
F/2-
Bol I ~
stn A2 ~
k tri A3 ~
stn A7 ~
Inst. 0."
0. IX-
0.S6X"
1.0%"
3.2%'
5.6%-
10%-
I
>rr» i
(111 l
LCELi
rcxn
tejn <
+
icjj I
\tfc <
i cjj <
3-H
0 200 400 600 680 1000 1200 1400 1600 1600 2000 2200 2400 2600 2800 3000
Thousand ceIl« p«r ml.
N Th» d 1 f f arenca Is sign If I conk Cby anova, P< 0.001); Ranga & moan + 2 S.E. are shown.
-------
5.6%-
10%"
Figure 9
DUNALIELLA C8th Day - 3 Oct 783
stn A2
stn A3
stn A7
Inst. 0.
0. I %
0. 56%
. 0%
3.2%
200
300
400
900
1000
500 600 700 800
Thousand ca1 Is par ml .
* Th« di ff«r«nc« Is «IgnIfI cant Cby anova, P< 0.0013; Rang* & mttan + 2 S.E.
! 100
1200
-J
00
<
>
cr*
shown.
-------
WILMINGTON
LONG BEACH
cc*
C7/
C3«
B4a
Pier J
D1»
09*
B3»
A15«
C2
A7
SAN x
PEDRO'
BID*
•CI
A12'
Long Beach Harbor
DS*
AO
A2*
A13*
B1«
Itlll K Mlllt
AO-
Figure 10. Estimated Zone of Phytoplankton Enhancement, 1978
Harbors Environmental Projects
University of Southern California
-------
480
VA 18

Table 1. 8-Day Mean Specific Growth Rates (aay ~)
Treatment
10%
5.6%
3.2%
1.0%
0.56%
0.1%
Stn Boil
Stn A7
Stn A3
Stn A2
Dilution
Water
Instant
Ocean
Enriched
Medium (F/2)
February
.49
.49
.49
.45
.44
.44
.42
.46
.43
.43
.44
.41
.42
April
.32
.29
.27
.21
.19
.18
.33
.20
.20
.17
.18
.19
.32
June
.32
.30
.28
.24
.25
.24
.19
.22
.23
.24
.24
.24
.31
August
.39
.36
.32
.25
.18
.08
.40
.29
.13
.11
.09
.07
.40
October
.28
.29
.27
.23
.24
.23
.29
.25
.19
.19
.22
.17
.26
-------
VA 19
Table 2. Nutrient Levels at Harbor Stations
481
N0„

Stations
Feb
April
July
Aug
Oct

TITP
Effluent
40
33
33
9
17

A7
.366
.166
.148
.440
.071

A4
.238
.102
.188
.114
.102

All
.263
.091
ND
.061
.127

A3
.271
.159
.210
.103
.065

A2
.174
.079
.120
.053
.077

A12
.143
.136
.082
.088
.056
N03







TITP
Effluent
921
921
64 3
64 3
500
500
93
93
1329
1329

A7
3.697
3.222
15.172
5.156
8.924

A4
1.980
.709
0
.244
3068

All
2.591
.747
ND
.385
1.858

A3
1.579
1.165
.191
.920
1.047

A2
2.379
1.023
.128
.478
1.466

A12
1.329
0
5.039
4.781
.372
NH3







TITP
Effluent
7001
(564)
995
(1343)
1291
(1093)
2500
(1100)
0
(0)

A7
35.388
10.253
8.005
119.279
1.654

A4
5.314
2.669
2509
1.976
.331

All
2.597
.971
ND
3.176
1.654

A3
8.032
4.186
1.912
4.941
.331

A2
5.979
1.941
1.553
3.176
1.433

A12
1.329
.789
1.195
*1.623
3.198
P04







TITP
Effluent
—
——




A7
9.523
3.456
.456
12.391
3.024

A4
1.249
1.216
.728
.971
1.105

All
1.594
.689
ND
1.689
1.295

A3
2.274
1.742
.631
1.362
.773

A2
1.792
1.304
.641
1.188
.706

A12
.761
.600
.697
.539
.453
1	2
Monthly mean	Week of bioassay
-------
Intentionally Blank Page
-------
VB 1
483
TERMINAL ISLAND TREATMENT PLANT SECONDARY WASTE BIOASSAYS
INTRODUCTION
The purpose of a bioassay test series is to determine the
effects of a particular substance on a group of selected
organisms. Short-term (96-hour) tests can reveal only acute
toxicity, whereas longer term tests (21 days or longer) are
needed to identify sublethal, or chronic effects. Substances
which may be contained within wastewater effluents discharged
into the marine environment are of particular concern. In
theory, if an effluent possesses a significant toxicity a con-
centration of this effluent can be found which causes a sig-
nificantly higher mortality to occur in the marine organisms
than does sea water not containing this effluent.
In the present study, the Terminal Island Treatment Plant
(TITP) effluent was investigated. The effluent consists of
10-18 million gallons of secondary-treated sewage waste water
released daily into outer Los Angeles Harbor. The original
experimental design employed the use of four species of
marine organisms which are common to the local nearshore waters
of southern California. These were used in five sets of bio-
assays over different seasonal time periods. In addition, one
other local marine organism was included in four of the bioassays
and an additional set of supplemental bioassays was performed.
EXPERIMENTAL DESIGN
To assess the toxicity of TITP waste water, 96-hour bio-
assays were performed with twelve different test solutions.
These solutions included six concentrations of sewage effluent,
four field samples of receiving water and two types of controls.
The concentrations of effluent used were 100, 75, 56, 32, 18
and 10 percent. The receiving water stations were located at
progressively greater distances from the TITP outfall. These
can be seen on the map, Figure 1, and were: 1) the TITP outfall;
2) at station A7, 550 m from the outfall; 3) at the Fish Harbor
entrance buoy designated A3, 1525m distant from the outfall;
and 4) the channel marker buoy A2, 1975m from the TITP outfall.
One control was a solution of "Instant Ocean", and the other
was filtered and ultraviolet-sterilized harbor water (desig-
nated henceforth as "house" water) from the USC Marine Facility
at Berth 186, Los Angeles Harbor.
The four species originally selected as test organisms were
Neanthes arenaseodentata (Polychaete worm), Aoartia tonsa
(planktonic copepod), Fundulus parvipinnis (California killifish)
and embryos of Engrculis mordax (anchovy). The additional
species employed in four of the five regular bioassays was
Emerita analoga (the sand crab).
-------
484
VB 2
The setup dates of the five regular 96-hour bioassays
were February 13, April 17, June 23, August 21 and September 25,
1978.	The supplemental bioassays were performed on January 29,
1979.
MATERIALS AND METHODS
Test Organisms
Test organisms were collected in various ways from several
different sources. The Fundulus were taken by fish trap from
a series of seawater canals in Venice, California. These fish
were acclimatized in flow-through holding tanks of "house"
sea water until the time of the bioassay setup. Emeviva were
sieved from the sand in the surf zone at Seal Beach. These
were held in flow-through aquaria with bottoms covered approxi-
mately two inches by clean sand. The polychaetes (Neanthes)
were readily available from laboratory cultures. On the morning
of bioassay test starts, Acartia and Engraulis embryos were
collected from plankton tows in Cerritos Channel and outside
the Los Angeles breakwater respectively. Once in the laboratory,
containers with these organisms were placed in water baths for
acclimatization with "house" sea water.
Experimental Setup
A regular bioassay setup included two lOgal. aquariums for
each of the twelve test solutions. This procedure was followed
in order to separate the Neanihes from the Fundulus, which will
predate upon them. These 24 aquaria were distributed amongst
five waterbaths which maintained the temperature of all bioassay
test solutions and controls at 17° C.
On the morning of the bioassay, secondary-treated effluent
was obtained from the sampling site at the Terminal Island
Treatment Plant. A 110 gal polyethylene tank was used to
transport the waste to the bioassay laboratory. Before trans-
ferring the waste water to the bioassay aquaria, the salinity
was adjusted with "Instant Ocean" to match the salinity of
"house" sea water.
In order to dilute the secondary waste to proper test con-
centrations, the salinity-adjusted waste water was poured into
appropriate test aquaria to a level predetermined for each
test concentration. Various quantities of "house" sea water
were then added to finish the filling procedure. In the case
of aquaria containing 100% test solutions, no addition of "house"
sea water was required. Similarly, no dilutions were necessary
in the aquaria containing control solutions or receiving water
samples; these were simply filled with the appropriate sea
water.
-------
VB 3
485
When all test containers had been properly filled, the
test organisms were distributed among them. Polychaetes were
removed from the laboratory culture aquaria and placed in
porcelain pans of "house" sea water. Since Neantkes can be
cannibalistic, it was necessary to keep the test individuals
separated within test aquaria. This was accomplished by
using isolation tubes composed of 1 inch by 2 inch cylindrical
snap-cup vials (polyethylene) whose lids and bottoms had been
replaced with Nitex screen to provide a freely exchanging test
chamber. One polychaete was loaded into each tube which was
then placed into a bioassay aquarium. Each of the aquaria
from one of the duplicate sets of twelve solutions received
20 housed individuals.
The Fundulus were placed into the other set of aquaria.
These were netted from their holding tank and ten individuals
were loaded into each of the appropriate twelve bioassay aquaria.
The fish were inspected prior to loading, and any with patho-
logical symptoms were discarded.
The Acartia and Engraulis were placed in separate plastic
beakers suspended in the set of bioassay aquaria containing
tleanthes. These beakers had screen windows in the sides to
allow free circulation of water throughout the beaker while
still retaining test animals. This method was intended to sim-
ulate better normal field conditions than does placing the
organisms in crystalizing dishes in a separate water bath, as
is usually done. The beakers allow the organisms to be in
contact with the test solution in the 10 gal. aquaria, which
are monitored daily for temperature, salinity, dissolved oxygen
and pH. Monitoring those parameters in crystalizing dishes was
impossible with available equipment.
A dissecting microscope was necessary for counting the
proper numbers of Aaartia and Engraulis embryos. Eyedroppers
were used to remove samples of Aaartia from their holding con-
tainer. The contents of the dropper were emptied onto a
depression slide and the copepods inspected and separated
under the dissecting scope. Healthy individuals (intact and
actively swimming) were removed with the eyedropper and trans-
ferred to the test containers. Twenty individuals were placed
into each of the containers in the twelve test solutions.
Anchovy embryos were separated in a similar manner. One-
day-old embryos which appeared healthy were selected and
transferred to the appropriate screened beakers. Thirty
individuals of Engraulis were used per test container.
When Emerita were used, they were screened from the sand
on the bottom of the holding aquaria and counted out into the
set of bioassay aquaria containing the Neanthes, Aeartia and
Engraulis . Twenty of these sand crabs were put into each test
solution.
-------
486
VB 4
The screened beaker method of containing Aaartia and
Engraulis within the large bioassay aquaria often resulted in
high mortalities. For this reason supplemental bioassays were
performed. Samples of the twelve test solutions were placed
in crystalizing dishes supported on racks in a water bath,
into which test organisms were added. This method resulted
in much lower mortalities among test organisms, but is less
scientifically pleasing, as monitoring of test conditions in
crystalizing dishes is impossible.
During the tests all aquaria were aerated to maintain the
dissolved oxygen level. This was necessary for the larger
test organisms;Neanthes, Fundulus and Emsrita. The lighting
cycle was 14 hrs. light/10 hrs. dark. None of the test organ-
isms were fed during the bioassay. The temperature, salinity,
dissolved oxygen and pH in each tank were monitored and
recorded daily.
After 96 hours, a final reading of the above parameters
was made and the test concluded. The surviving Fundulus,
Emerita and Neanthes were counted by direct inspection while
the organisms were still in the test aquaria. A dissecting
microscope was used to count Aaartia and Engraulis embryos
and only live organisms were tabulated. Normally, these test
results for all solutions and test species were tabulated
within four hours.
All aquaria, plastic containers and glassware used in
these bioassays were pre-washed with tissue-grade detergent
and 10 percent hydrochloric acid. This procedure was followed
to prevent contamination from previous bioassays.
RESULTS
The bioassay test results are presented in the six tables,
1 through 6. In these, the percent survival of test species
is presented for all twelve test solutions and six 96-hour
test periods. Occasionally, mortalities were known to be due
to escaped animals and when possible these are noted.
Also included are the means of the daily recordings of
measured experimental conditions; temperature, salinity,
dissolved oxygen and pH. These are presented below the percent
survival data with their standard deviations. The standard
deviations are very small in all cases and it is judged by the
experimentors that these parameters had negligible effects on
the survival of test organisms.
-------
VB 5
487
DISCUSSION AND CONCLUSIONS
Two of the species employed in this series of bioassays,
Fundulus (the California killifish) and Neantb.es (a polychaete
worm) showed no apparent difference in percent survival during
any bioassay over the total range of test solutions. Survivor-
ship was always near 100% for both of these species in all
secondary waste dilutions, all receiving water samples and in
both types of controls. A third species, Emerita, showed this
same trend in three of the four bioassays in which it was used.
The results indicate that the TITP effluent has no inherent
toxicity for these organisms, even in the extreme case of the
100% concentration of secondary waste.
The Aaartia results also support this conclusion and do
not seem to show any differential survival in the various test
solutions for different test dates. However, extremely low
survivorship of these organisms in all solutions makes it
difficult to draw conclusions from the data.
Engraulis data from the supplemental bioassays performed
on January 29, 1979 mirror the Fundulus and Neanth.es results.
Here there were no significant mortalities in any test solu-
tion or for any of the four replicates per solution. However,
the anchovy embryos did not maintain this trend on other
bioassay test dates. The high survival during the January test
may have been due only to the use of different techniques. The
Engraulis were placed in crystalizing dishes in a water bath
rather than in screened beakers suspended in test aquaria.
The remaining combinations display a different trend. This
is towards a perceptibly higher mortality in high concentrations
of effluent than in the control groups. This occurred with
Emevita on April 17 and with Engraulis on February 13, April 17
and June 23, 1978. In these Engraulis bioassays, the screened
beaker method, as described in Materials and Methods, was
employed. As was mentioned there, this method was used to
allow monitoring of experimental conditions within the Engraulis
test containers. This method resulted in high overall mortal-
ities in all solutions. This high mortality implies that these
organisms were stressed to a greater extent than they would ever
be in nature and would be more susceptible to toxic effects
than in a natural situation.
Thus, conclusions of toxicity of the TITP effluent would
be exaggerated. A similar situation may have occurred with
Emerita in the April 17 test, as this species shows no signifi-
cant mortality in any solution in any other test.
These inconsistent results between different test periods
may also reflect variations in the character of the TITP waste-
water effluent over time. In this case, receiving waters may
-------
488
VB 6
be toxic to more delicate organisms, such as Engraulis embryos,
during certain periods when the effluent discharged contains
some particular toxic substance. This could occur while the
same effluent waters remain innocuous to more hearty species
such as Neanthes and Fundulus.
Wild harbor Aoavtia populations have not proven successful
in bioassay tests for approximately the last two years. Condi-
tions in the preferred inner harbor habitat may have changed
to stress the populations so that they are less able to tolerate
the capture and testing than previously. Numbers have been
greatly reduced at times recently.
In general, one could conclude that if toxic effects
should occur from the discharge of TITP effluent, they would
be related to wastes introduced that were not there during the
present tests.
-------
SCAte IN Milts
WILMINGTON
LONG
BEACH
TITP
SAN
PEDRO
>3,
CABRILLO
BEACH/
FIGURE 1
TITP BIQASSAY/TOXICITY
EFFLUENT SAMPLING STATIONS
<
W
-J
00
U3
-------
490
VB 8
TABLE 1 • TEST 1 - FEBRUARY 13, 1978
CONCENTRATION
OF EFFLUENT
PERCENT SURVIVAL
CONTROL
10%
18%
32%
56%
75%
100
FUNDULUS
100
100
100
100
100
100
100
NEANTHES
100
90*
100
100
100
100
100
ENGRAULIS
90
67
97
100
50
53
13
ACARTIA
0
0
0
0
0
0
0
RECEIVING WATER
station
BOIL	100	95*	0	0
A 7	100	100	60	0
A3	100	100	80	0
A 2	100	90*	87	0
*THESE APPARENT MORTALITIES ARE DUE TO ANIMALS MISSING FROM THE
TEST CONTAINERS. THEY MAY HAVE BEEN EATEN, OR NOT SEEN UPON
COUNTING.
EXPERIMENTAL CONDITIONS
TEMPERATURE
15.6
+
0.37 °C
SALINITY
33. 1
+
0.26 PPT.
DISSOLVED OXYGEN -
8. 1
X
0.2 mg/I
PH
8.0
+
0.2
-------
VB 9
491
TABLE 2. TEST 2 - APRIL 17, 1978
CONCENTRATION
OF EFFLUENT
PERCENT SURVIVAL
INSTANT OCEAN
CONTROL
LAB WATER CONTROL
10%
18%
32%
56%
75%
100%
FUNDULUS
100
100
100
100
100
100
100
100
NEANTHES ENGRAULIS ACART IA EMERITA
90
100
100
100
100
100
100
95*
50
23
53
33
50
3
10
3
0
3
7
0
0
3
0
0
100
100
100
95
95
90
75
0
RECEIVING WATER
STATIONS	
BOIL
A7
A3
A2
100
100
100
100
100
100
1 00
100
0
0
0
1 0
3
0
0
3
95*
100
100
90*
* THESE APPARENT MORTALITIES ARE DUE TO ANIMALS MISSING FROM THE
TEST CONTAINERS. THEY MAY HAVE BEEN EATEN, OR NOT SEEN UPON
COUNTING.
EXPERIMENTAL CONDITIONS
TEMPERATURE
- 17.1
+
0 . 3
°c
SALINITY
- 33.6
+
0 . 5
PPT
DISSOLVED OXYGEN
- 7.3
+
0 . 6
mg/1
PH
- 7.8
+
o
•
o
1
-------
492
VB 10
TABLE 3.
TEST 3 - JUNE 23, 1978
CONCENTRATION
OF EFFLUENT
PERCENT SURVIVAL
FUNDULUS
INSTANT OCEAN
CONTROL	100
LAB WATER CONTROL	100
10%	100
18%	100
32%	90
56%	80
75%	100
100%	90
NEANTHES ENGRAULIS ACARTIA
75
100
95*
100
100
95*
95
95
40
25
15
25
20
10
0
0
0
10
25
20
15
0
0
0
EMERITA
100
100
100
100
100
100
100
100
RECEIVING WATER
	STATIONS	
BOIL
A 7
A3
A2
100
100
90
100
95*
100
100
100
15
0
5
0
50
20
65
10
100
100
100
100
~THESE APPARENT MORTALITIES ARE DUE TO ANIMALS MISSING FROM THE
TEST CONTAINERS. THEY MAY HAVE BEEN EATEN, OR NOT SEEN UPON
COUNTING.
EXPERIMENTAL CONDITIONS
TEMPERATURE
- 19.5
+
1 . 2
°C
SALINITY
- 32 .8
+
0.7
PPT
DISSOLVED OXYGEN
- 7.7
+
0.5
mg/1
PH
- 7.5
+
0.2

-------
VB 11
493
TABLE 4. TEST 4 - AUGUST 21, 1978
CONCENTRATION
QF EFFL.V£NT
PERCENT SURVIVAL
FUNDULUS
INSTANT OCEAN
OCEAN.	100
LAB WATER CONTROL	100
'10%	100
18%	100
32%	100
56%	100
75%	100
100%	90
NEANTHES ENGRAULIS ACARTIA
100
100
95
100
100
100
100
100
43
0
17
7
10
0
0
0
20
10
27
0
0
0
7
0
EMERITA
100
100
95
100
100
95
95
100
RECEIVING WATER
STATIONS
BOIL
A 7
A3
A 2
100
90
100
90
100
100
95
100
10
27
10
0
13
0
0
7
100
100
100
1 00
~THESE APPARENT MORTALITIES ARE DUE TO ANIMALS MISSING FROM THE
TEST CONTAINERS. THEY MAY HAVE BEEN EATEN, OR NOT SEEN UPON
COUNTING.
EXPERI MENT A1 CONDITIONS
TEMPERATURE!
- 17.9
+
0.3
°c
SAL IN ITY
- 32.0
+
0 . 6
PPT
DISSOLVED OXYGEN
- 9.6
+
0 . 7
mg/1
PH
- 8.2
+
0. 1

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494
VB 12
TABLE 5. TEST 5 - SEPTEMBER 25, 1978
CONCENTRATION
OF EFFLUENT
PERCENT SURVIVAL
INSTANT OCEAN CONTROL
LAB WATER CONTROL
10%
18%
32%
56%
75%
100%
FUNDULUS
100
100
100
100
100
100
100
100
NEANTHES
90
95
90
100
90
100
100
100
EMERITA
100
100
95
95
100
95
1 00
95
RECEIVING WATER
	STATION?
BOIL
A 7
A3
A2
100
100
100
100
85
90
100
100
100
100
85
100
EXPERIMENTAL CONDITIONS
TERMPERATURE
- 20.8
+
1.0 * C
SALINITY
- 31.0
+
0.4 PPT.
DISSOLVED OXYGEN
- 9.3
+
0.4 rag/1
PH
- 8.1
+
0.1
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TABLE 6. SUPPLEMENTAL TEST - JANUARY 29, 1979
CONCENTRATION
OF EFFI UENT	
PERCENT SURVIVAL
INSTANT OCEAN
CONTROL
LAB WATER CONTROL
10%
18%
32%
56%
75%
100%
RECEIVING WATER
STATIONS	
NEANTHES
93
93
93
93
93
97
93
90
ENGRAULIS-1
83
100
90
97
93
97
9 3
100
ENGRAULIS—2 ENGRAULIS — 3
ENGRAULIS-3
100
95
90
100
100
100
100
100
100
95
90
100
90
9b
95
85
95
95
95
100
100
ACARTIA
1 3
6
50
60
30
60
33
67
<
ffl
CJ
BOIL
A 7
A3
A 2
93
97
100
100
90
97
1 00
90
90
100
100
85
85
90
100
90
85
95
95
85
77
60
80
87
EXPER IMENT AI	CONDITIONS
.to.
TEMPERATURE	-12.3+0.1 °C	^
SALINITY	- 30.7 +. 0.9 PPT
DISSOLVED OXYGEN - 8.5 1 0.6 mg/1
PH	-8.2+0.1
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496
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vc
497
CANNERY WASTE AS A POOD FOR ANCHOVIES
INTRODUCTION
In September of 1978 a report was prepared on the results
of experimental feeding of anchovies on wet sludge obtained
from the StarKist dissolved air flotation (DAF) treatment of
fish cannery waste (Ralston, private report, 1978). In that
experiment, one group of fish received supplemental feeding
with sludge while the control group did not. The only food
common to both sets of fish was the ambient plankton, contained
in the water from Berth 186 in inner Los Angeles Harbor, that
was being pumped continuously into their tanks. Under that
regime, statistically significant differences were found in
mortality rates. The fish that received supplemental feeding
showed greater survival. However, growth curves showed that
both groups lost weight, indicating that the amount of sludge
selected for feeding was not sufficient to maintain the popu-
lation. Feeding levels ^^ere on the conservative side because
excess food in the tanks would cause bacterial problems for
the anchovies. Although the weight loss was less in the fish
receiving the sludge as a supplement, the results were not
statistically significant. Mortality is always high in
captive anchovies, and it is important to note that mor'talities
were significantly fewer in the sludge-fed group.
The present experiment was modified to present a higher
level of overall feeding and was designed for better statis-
tical analysis.
METHODS
Eight tanks containing 60 anchovies each was fed a main-
tenance diet of 15 grams of trout chow per day. Six of the
tanks in replicate pairs were given 5, 10 and 15 grams per
day of dried, ground sludge as a supplemental ration, respec-
tively. The remaining pair of tanks received no supplement
and served as the controls. All tanks were served with
continuously pumped harbor water, as was the case in the
earlier experiments.
Prior to the start of the present experiment, each fish
was weighed and the average weight of the fish in each tank
was calculated. At the end of 15 days the individual fish were
again weighed and the average weight was determined for the
surviving fish in each tank. The average fish weight per tank
was used rather than average weight per fish to correct for
the normal mortality of captive anchovies during the course of
the experiment.
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498
VC 2
RESULTS
Mortality, although high, averaged 41% and was not sig-
nificantly different from tank to tank. The average weights
in each tank at the start of the experiment, the average
weights at the end, and the differences are shown in Table 1.
A linear regression analysis was performed on these net
growths. The results are shown in Table 2 and plotted in
Fi-gure 1. The increase in net growth fits a rising straight
line (Fs for a straight line of slope not equal to zero yields
P<.05). Deviations from the straight line model are insignifi-
cant (Fs yields P>.8).
The fish receiving the maximum amount of sludge got twice
the weight of food {15 gms sludge + 15 gms trout chow) as the
control fish (15 gms trout chow only); yet their growth was
triple that of the control (about .27 gms increase compared
with about .08 gms increase in the controls). In this experiment,
as in the previous one, the saturation point or maximum amount
of sludge that can be utilized for growth was not reached.
DISCUSSION AND CONCLUSION
It is valid to conclude that the anchovies can utilize the
sludge for growth. However, another set of experiments would
be needed to examine the upper limits of the growth curves.
The two sets of experiments have indicated that the wet or dry
sludge is supportive of growth for anchovies. However, the
California Department of Fish and Game and the Environmental
Protection Agency have not permitted disposal of the sludge
from the Terminal Island canneries at sea. Instead, this
nutrient source is being dumped in a landfill. When the sludge
is wet, it creates odors as microbial biodegradation occurs.
Drying the sludge is energy demanding and adds to waste disposal
costs, which now include sewerage of the liquid wastes.
Questions raised previously on metals content of sludge
are being addressed by the canning industry. The Environmental
Protection Agency reiterates that containment of sludge on
land, with attendant odor and leaching probabilities, is better
for the environment than dispersion into open waters of the
ocean. This appears to constitute a wasteful solution for a
nutrient source, which has potential for a mariculture nutrient
or for feeding natural marine populations. Feeding sludge to
pigs is also being tested elsewhere.
LITERATURE CITED See Section VI
-------
VC 3
499
Table 1. Average weights and net change in weight of fish
fed on different supplemental rations of sludge
gins
sludge
replicate
average
start
fish weight
end
weight
increase
0.0
1
2.04
2.21
0.18
0.0
2
2.53
2.51
-0.02
5.0
1
2.16
2.25
0.09
5.0
2
2.10
2.25
0.15
o
o
r-H
1
2.29
2.47
0.18
10.0
2
2.02
2.21
0.18
15.0
1
2.08
2.39
0.32
15.0
2
2.27
2.50
0.23
Table 2.
LINEAR REGRESSION ANALYSIS
ANCHOVY FEEDING STUDY
X represents grams sIudge in food per day.
Y represent* growth of average fish (grows).
For 8 points suppIied, tho moan of X Is 7.5000.,
and the mean of Y is 0 1627.
The vor i arico of X i s 35 7143 and of Y is 0.0097.
The regression equation i s Y= 0.0129 X +¦ 0.0660,
95.0% confidence limits for tho slope are 0.0056 & 0.0203.
ANALYSIS OF VARIANCE-
SOURCE SS
gr oupc
' i neor
dev .
error
tota I
0.0431
0.0416
0.0015
0.0245
0.0676
df
3
4
7
1
2
MS
0.0144
0.0416
0.0007
0.0061
2.3467
57.0813
0.f192
0.214
0.017
0.891
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ANCHOVY FEEDING STUDY
U1
o
o
0.4
0.3
0.2-
0 I
-0. 1
cIudgo
grams
3
Figure i. The regression line and 95% confidence limits are plotted with the range
AND 2 S.E.'S OF THE ORIGINAL DATA AT EACH X VALUE.
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V Dl
GROWTH AND STIMULATION OF INVERTEBRATES
IN THE WASTE PLUME
501
BIOSTIMULATION OF MYTILL'S EPULIS
INTRODUCTION
Biological laboratory studies cannot simultaneously
reproduce the synergism of physical and chemical factors which
occur in natural ecological systems. For this reason, it is
desirable to augment laboratory studies with actual in situ
biological experiments. In this study, in situ growth experi-
ments help to assess the impact or biostlmulatory effect of
Terminal Island Treatment Plant (TITP) wastes on the marine
environment.
The bay mussel, Mytilus edulis, is a common fast-growing,
filter-feeding mollusc, which occurs in all semi-protected
waters of southern California, as well as in many other areas
of the world. Any hard substrate which is not subject to
periodic artificial disturbances and which lies within the
tidal levels of approximately +1 to -3 meters from mean
lower low water, is usually encrusted with mussel growth.
The metabolic potential of these dense mussel beds is signifi-
cant to the ecological balance in coastal marine waters. For
these reasons, Mytilus edulis was chosen as an indicator
species for determining the growth potential of organisms
affected by the TITP wastewater outfall.
EXPERIMENTAL DESIGN
TITP effluent has a substantially lower salinity than
normal harbor waters. This results in a lower density and a
tendency of the effluent to form a surface lens, which becomes
less distinct and more mixed with increased distance away
from the outfall. Complex harbor circulation patterns also
affect the horizontal distribution and mixing of TITP waste
waters. In order to take into account this three-dimensional
effluent distribution, a vertically — as well as horizontally —
stratified sampling scheme must be employed.
In this experiment, three depths were designated for sus-
pending samples of mussels. These were one meter, two meters
and three meters deep. These depths were represented at four
stations at various distances from the TITP outfall during
each of four one-month experimental periods. Station locations
were: 1) at the TITP outfall, 2) at the buoy designated A7,
550 m distant from the outfall, 3) at the Fish Harbor entrance
buoy designated A3,15 25 m from the outfall, and 4) at the
channel marker buoy A2, 1875 m distant from the outfall (see
Figure 1). The experimental periods were: 1) May 18-June 18,
-------
502
V D2
1978, 2) August 9-September 9, 1978, 3) October 24-November
24, 1978, and 4) December 5, 1978-January 5, 1979.
Each sample initially consisted of 40 mussels. This
sample si2e was selected so as to allow for some natural mor-
tality and still be large enough after one month for optimal
statistical analysis. Any mortalities observed were deter-
mined to be mainly due to predation and unrelated to effluent
concentrations. Each mussel in the samples was measured to
an- accuracy of .005 cm at the beginning and end of the test
period, in order accurately to determine individual mussel
growth.
MATERIALS AND METHODS
In nature, Mytilus edulis occurs in dense clumps in asso-
ciation with many other fouling organisms. In this complex
association, Mytilus edulis compete with the other fouling
organisms, as well as with each other, for filterable food
particles in the surrounding water. These complex interactions
in intact mussel clumps make it impossible to determine
accurately growth potentials for individual mussels main-
tained in receiving waters.
To alleviate this problem, mussels were cleaned of all
fouling organisms and suspended separately from each other.
This was accomplished using specially designed and constructed
mussel racks. The racks also permitted keeping track of
individual mussels, so that monthly growth was determinable
for each individual. The determinations greatly augmented
statistical analysis of the data.
Mussel Racks
Figure 2 is a diagram representing one of the four mussel
racks. Each rack consisted of three h inch thick stainless
steel hoops arranged in a vertical array at one meter, two
meter, and three meter depths. Each hoop was connected to
the next one by three 3/16 in diameter stainless steel cables.
Stainless steel thimbles and "nico-press" fittings were used
to attach cable ends to prevent chafing. Racks were held
away from buoys by rigid stainless steel struts guyed to
buoy chains by stainless cables. Stainless snap-shackles were
used to attach the support struts to a bridle extending from
the bottom hoop of the rack. These facilitated quick attach-
ment and removal of mussel racks. Racks were supported
vertically in the water column between the rigid support strut
and a submerged high impact plastic float attached to the top
hoop. This arrangement was designed to always keep the mussels
at their respective test depths and to circumvent vandalism.
-------
V D3
503
A folded-over 6 inch strip of h inch stretched mesh,
knotless nylon netting was sewn around the circumference of
each hoop. Each folded strip contained 40 individual heat-
sealed, numbered pockets to hold each of the 40 individual
test mussels per hoop. Once a mussel was placed into a
pocket, it remained there throughout the month-long experi-
mental period. At the end of the period, each could be
removed, remeasured, and the final length compared to its
initial length to determine individual mussel growth. Since
this arrangement maintained mussels separately from one
another and away from other fouling organisms, competition
with other filter-feeders was minimized. All of the 40 mussels
at a particular depth and location should have had an equal
opportunity to feed during any given experimental period.
Loading of Mussels
All mussels used in this experiment were collected from
the same group of pilings marking the east side of the main
channel entrance to inner Los Angeles Harbor. These pilings
lie approximately 100 meters offshore in about 60 feet of
water. This site receives more surge from passing ships than
most harbor areas, resulting in mussel growth with minimal
fouling by other organisms. These relatively "clean" mussels
were ideal for this experiment as they required little initial
cleaning prior to measurement and loading into mussel pockets.
Mussels were always collected at low tide a few days prior to
the start of an experimental period and held in running sea
water in the laboratory until used.
Mussels were loaded into their respective pockets on the
mussel racks one day before field deployment. The procedure
incolved the random selection of an individual mussel from a
holding tank; the careful removal of fouling organisms off
the mussel; measurement of the maximum length of the shell's
long axis to the nearest 1/20 mm (with an outside vernier
calliper); the recording of this measurement; and finally, the
section of the mussel into its respective nylon net pocket on
the rack. When a mussel rack was fully loaded, it was stored
in a 500 gallon holding tank with flowing sea water until
deployment in the field the following day. After one month
in the field, racks were returned to the lab, the mussels
individually removed from their pockets and remeasured as
before. Mussels were discarded after use and new ones obtained
for each of the four one-month experimental periods. The
purpose of this was to prevent any residual effects of one
test from influencing the outcome of other tests.
-------
504
V D4
RESULTS
Over 3,500 mussel measurements were taken from three
depths, four locations and four one-month experimental time
periods. This mass of data was computerized and analyzed
statistically to determine whether any significant differ-
ences occurred between any of the experimental parameters.
Summary of Analysis
Figures 3 and 4 and Tables 1 and 2 are the results of
two regression analyses performed to determine whether absolute
length increase or per cent length increase should be used
for the analysis. A subset of 9 6 individuals was drawn from
the data for this analysis (two randomly selected from each
of 48 samples). Within this subset, absolute length increase
remained constant over the entire range of the starting sizes
(that is, the slope of the regression line was not significantly
different from zero), so may be assumed not to be a function
of size. This was not true for per cent length increase, which
fell off with size increase. As a result, absolute length
increase was used for all remaining analyses.
This experiment fits a model I, factoral design ANOVA
(three factors with replicates). The analysis was performed
after the manner of Hartley (19 62). This method requires a
balanced model. Since there was missing data due to mortality,
the smallest sample was used as the replicate size (17 individ-
uals) , and individuals were randomly eliminated from the other
samples to bring them down to the balanced sample size.
In the Hartley method, the replicates are considered as
a fourth factor. Following computation, all terms containing
the "replicate factor" (factor A) are pooled to form the error
term. The results of the Hartley method are shown in Table 3,
and the pooled ANOVA in Table 4.
Only the variations due to season and location and their
interaction were significantly greater than the overall growth
variation. This conclusion is supported by Figures 5, 6 and 7,
in which the means, ranges and two standard error boxes for
each factor (all individuals pooled) are plotted. The standard
errors overlapped among depths, whereas they did not among
locations and seasons.
Figure 8 presents the interactions of season and location.
The growth with season is plotted for each test location. It
appears that the increased August to November, 1978 growth
seen at A7 and A3 was enhanced by the TITP outfall. Racks at
station A2, however, appeared to have reacted differently, for
unknown reasons. The nutrient supply may have been lower at
A2.
-------
V D5
505
DISCUSSION AND CONCLUSIONS
Among the factors investigated that might be related to
growth rate of Mytilus edulis in the harbor it was found that
depth and size at the start of the experiment were not signifi-
cant. Season and proximity to the outfall were statistically
significant.
As shown in Figure 8, the seasonal growth rates for mussels
held for one month at the TITP boil and at stations A7 and A3
followed similar trends. Low rates of growth were found in
May-June, 1978 and December 1978-January 1979. Higher values
were found for the two experimental periods between those dates.
The station at the boil consistently showed the highest growth
rates of the three and station A7 the lowest.
Station A2, the farthest from the outfall area, showed
highest growth rates of all stations in the spring (May-June
1978} and winter (December 1978-January 1979) . During the summer
and fall periods, when the other stations showed enhanced growth
rates, this station showed reduced growth rates, the lowest
among the four stations.
The trends in the seasonal growth rates clearly fall into
two categories. Stations at the TITP boil, A3 and A 7, are those
that are influenced by the wastes discharged from TITP into that
area. Station A2 is the farthest from the discharge area and
the closest to the open sea. Growth rates exhibited by mussels
suspended there probably reflect more the influence of the
ocean waters rather than the effluent from the treatment plant.
It is interesting to speculate on the relationship which
the growth rate curves may have to the suspended solids and
other material in the TITP effluent rather than to seasonal
factors. Mussels are filter feeders, whose growth may depend
on the concentration of food particles in the surrounding waters.
It is known that particulate matter was copiously discharged
during the summer of 1978, when the treatment plant was upset.
The growth rates of the mussels rose at this time, when the
growth rate at A2, reflecting oceanic influence, dropped. The
higher fall growth rates may be a reflection of the same
influences governing the higher values at A2. The similarity
does not hold for the winter values.
At the TITP boil, higher oxygenation from the plant and
from the turbulence may account for the much higher growth
level in the summer than at station A7, the next closest loca-
tion. Nutrient levels would not be greatly different between
the two sites. At station A3, circulation is also probably
better than at A7 because of its less sheltered location. The
drop in growth at A2 is unexplained, except that A3 is generally
higher in nutrients such as N02, N03 and NH3. It is possible
-------
506
V D6
that some upwelling occurred during the winter period outside
the harbor, that would have brought mutrients into the main
channel on tidal exchange.
BIOSTIMULATION OF INVERTEBRATES
INTRODUCTION
The water column of marine areas with polluted or uncon-
solidated bottom sediments may be richer in fauna than is indi-
cated by bottom (benthic) sampling, and zooplankton tows
capture only small samples in time and space. There are many
organisms that are temporarily represented in the plankton as
eggs and larvae, which settle out when suitable substrates are
available, but otherwise perish. The settling rack technique
offers an artificial substrate, suspended from buoys and docks
at 3m depth. Results of the 1973 and 1974 studies were discussed
in AHF (1976).
In the usual harbor monitoring, racks are exposed for one
month periods. Fauna so collected differ greatly in space and
in time in the harbor. Therefore, in the present study, racks
were all exposed for one month at a single location, and then
transferred to separate sites for evaluation of further growth
during the second month.
METHODS
Quantified samples of one month old settling communities
were transplanted to four locations on a transect from the TITP
boil to the A2 channel marker buoy in the outer Los Angeles
Harbor. Analysis of these samples allows a good comparison of
the in situ growth and recruitment characteristics of settling
organisms throughout the TITP effluent plume.
The substrate for recruitment and growth of settling com-
munity samples was provided by settling racks developed by
Dr. John Soule at USC (Soule and Soule, 1971). These racks
consist of paired, open, wooden microscope slide boxes suspended
vertically from a single wooden cross support by 5/16 inch
nylon line. Twenty-five glass microscope slides are inserted
into slide slots in each of these boxes, which are then covered
with plastic screen, providing protected internal settling
surfaces in addition to the external surfaces which are exposed
to normal wave and current conditions.
The paired settling racks were first soaked in filtered
and ultraviolet-sterilized sea water for two weeks in the
laboratory. This procedure prepared the settling surfaces by
leaching out any toxic or inhibitory chemicals from the wood
and glue used in construction of slide boxes. In addition,
-------
V D 7
507
this allowed an accurate determination of the wet weight of
settling racks prior to the accumulation of any settling biomass.
The racks were weighed on a grocery scale to the nearest h ounce
the day of initial deployment in the field.
On June 14, 1978 four pre-soaked and weighed settling racks
were deployed at the A2 channel marker buoy at a depth of two
meters. This station was selected as the control site from
which the one-month-old settling community samples would be
acquired. It is approximately 1875 meters away from the TITP
boil towards the Angels Gate entrance to the Los Angeles Harbor.
Selection of this station as a control site is justified by
past hydrographic evidence (Robinson and Porath, 1974}, suggest-
ing a minimal effect at this location from the TITP effluent plume.
One month later, on July 14, 1978, the four settling racks
were recovered from the A2 channel marker buoy, wet-weighed
immediately on a grocery scale to the nearest h ounce and
photographed close up on both sides with a 35mm Canon AE-1
camera. The slides produced from these photographs were used
to determine general species composition of the settling com-
munities prior to deployment at test sites. Care was taken in
handling of the settling racks so as to keep the fauna alive
by minimizing air exposure of settling organisms and other
related physiological stress.
Four test locations were selected for the deployment of
the one-month-old settling community samples obtained on the
settling racks. These were: The control station at the A2
channel marker buoy stations, A3, A7, and directly at the TITP
effluent boil. The distances from these first three stations
to the TITP boil are approximately 1875 meters, 1525 meters and
550 meters respectively. All racks were resuspended at the
previous depth of 2 meters.
On August 14, 1978, one month after deployment at test loca-
tions, all settling racks were recovered. Weights and photo-
graphs were obtained in the same manner as on July 14, 1978, and
all four settling racks were preserved in 10% formalin solution
for subsequent laboratory analysis of species composition and
numbers.
RESULTS
Biomass of Settling Organisms
Data obtained from live weight measurements of intact
settling racks are presented in Table 1, in which the initial
weights of settling racks before field deployment, as well as
the weights after one and two months in the field, are given.
The net biomass weight of organisms has been determined from
-------
508
V D8
the gross weights. This was accomplished by subtracting the
initial wet weight of laboratory-seasoned settling racks from
the weights after one and two months respectively. These
biomass values are also presented in Table 1.
With these data, the percent increase in biomass of these
sample settling communities can be determined. This is cal-
culated by subtracting the first month biomass for a given
location from the biomass found at the end of the second month/
and then dividing this difference by the first month biomass
again.
% increase	2nd month biomass - 1st month biomass
in biomass	-j_gt month biomass
The values for the percent increase in biomass are also
given in Table 5 for each of the four settling rack locations.
These values have been represented graphically in Figure 9 to
show how the percent biomass increase of these settling
organisms relates to distance away from the TITP wastewater
outfall.
Species Composition
The number of species (or taxa) on the racks was increased
by the two-month exposure and double racks, so that direct
comparisons with one-month single racks would be misleading.
In general, there were 13 more taxa at A3 in the experi-
ment than from a comparable period one-month single rack
exposure,and 24 more taxa at A2 than on a single one-month
rack in August. The principal differences in space were that
station A7 had the highest number of taxa but fewer phyla (or
equivalent level). The differences are slight, however,
between A7 racks and A2 in numbers, but A2 had more phyla.
Text Table 1. Comparison of Species/Taxa on
Settling Racks
Number
Boil
A7
A2
A3
species/taxa
40
47
46
41
phyla
10
9
11
10
CONCLUSIONS
In the set of experiments,the percent biomass increase was
greater at station A7, as compared with the TITP boil. Station
A2 racks had the highest percent increase, while station A3
racks were the lowest. The increase at the lowest, however,
was still nearly 100%. Since A2 racks remained at the same
-------
V D9
509
site for the entire period, they would possibly have had an
advantage in not being transferred to a different regime.
Contrary to the concept that increase in biomass is traded
for reduced numbers of species or taxa, A2 and A 7 had the
highest and second highest percent increase in biomass
respectively, whereas they were almost identical in having
the highest number of taxa. Station A2 had the most phyla
of the four, but the differences are probably not significant.
It is clear that the TITP effluent plume is not inhibiting
the growth of water column invertebrates, but is providing
nutrients to a food chain which enhances growth.
FLOW-THROUGH BIQENHANCEMENT STUDIES OF THE TERMINAL ISLAND
TREATMENT PLANT SECONDARY WASTE EFFLUENT
INTRODUCTION
The Terminal Island Treatment Plant (TITP) effluent is an
important nutrient source, as is shown in the studies on
Mytilus edulis, settling rack invertebrates, and Phytoplankton
in receiving waters. The present study was designed to carry
these investigations further and to assess the bioenhancement
potential of the TITP effluent in a totally simulated labora-
tory ecosystem.
Two main questions are investigated here. The first is:
Can growth occur in selected species from this simulated eco-
system during long-term enrichment of the TITP waste water?
The second question is: Can the ecosystem purify these
simulated receiving waters biologically to make them more
esthetically pleasing in compliance with water quality criteria?
MATERIALS AND METHODS
a. Simulated Ecosystem Growth
In the first part of this experiment, the question of
whether TITP effluent can support growth in a simulated eco-
system is investigated. For this, a highly nutrient-rich com-
ponent of the TITP waste was used. This was pre-DAF-treated
fish cannery waste water and represented a large volume
component of(and a large percentage of the BOD contained in)
the TITP waste influent.
Approximately 1,000 gallons of this pre-DAF cannery waste
was trick]e-fed into the experimental setup during the 1% month
test period from November 7, 1977 to December 22, 1977. The flow
rate was maintained at approximately 100 ml/min.
-------
510
V DlO
The experimental setup consisted of four eight-foot-long
fiberglass troughs arranged in pairs, so that the outflow of
the first in a pair would flow into the second. One of these
separate two-trough systems was designated as the test set to
which the DAF cannery waste was fed and the other was a control
which received only laboratory sea water.
Into each of these sets of troughs was loaded 100 weighed
and measured clams Meroenaria meroenaria. Each week these
clams were rearranged within their respective troughs to
provide uniform feeding conditions for all clams.
Ten specimens of Fundulus parvipinnis, the California killi-
fish, were also used per set of troughs. These were also weighed
and measured prior to loading. In addition, a third Fundulus
group was set up in the laboratory, which was fed their usual
diet of trout chow.
Other organisms were included in these troughs, such as
the algae Enteromorpha and the tectibranch mollusc Aplesia
aalifornioa, but only the Meraenaria and Fundulus were quantified.
b. Biological Purification of TITP Effluent
In the second part of the study, a simulated mussel bed and
phytoplankton ecosystem was tested to determine its ability to
purify waste waters biologically. The TITP secondary waste
effluent was used in this study. This effluent was first diluted
to 32% with Harbor Laboratory sea water, inoculated with a sample
of harbor phytoplankton and allowed to incubate in the sun until
the plankton reached a thick "bloom" stage. The initial setup
date was March 16, 1979 and the phytoplankton culture was
trickle-fed to the test troughs on March 28, 1979. This allowed
12 days for the phytoplankton to utilize the nutrients within
the TITP waste water solution.
Four sets of two 8-foot-long troughs each were used in this
experiment and set up in a similar manner as in Part a. Into
these, a layer of clumps of the mussel Mytilus edulis was then
added, which contained the barnacle Balanus sp., the green alga
Enteromorpha sp., the nudibranch Hermisenda sp., the anemone
Anthropleura sp., as well as many unidentified polychaetes,
flatworms, tunicates, hydroids and marine organisms commonly
found in mussel associations. In addition, 50 clams (Mercenaries,
meraenaria) were added to each lower trough.
Flow rates were calculated for each of the four sets of
troughs to give sewage effluent concentrations of 0 (control),
1.35, 5.6 and 10 percents. A total of 9.9 L/min of flow was
maintained in each set of troughs. To maintain the above
effluent concentrations, while using the stock TITP fffluent
solution of 32 percent effluent/68 percent laboratory sea water,
-------
V Dll
511
varying proportions of TITP effluent to laboratory sea water
were used to make up the total 9.9 L/min flow. The control
set had no sewage solution and 9.9 L/min laboratory sea water,
concentration #1 had 0.4 L/min sewage solution and 9.5 L/min
laboratory sea water, concentration #2 had 1.7 L/min sewage solu-
tion and 8.2 L/min lab sea water, and concentration #3 had
3 L/min of sewage solution and 6.9 L/min of lab sea water. These
flow rates were monitored throughout the experiment with pre-
cision flow meters and adjusted when necessary with individual
"ball" type PVC valves.
The total valume of the sewage solution in the phytoplankton
culture- aquaria was 20 36 L. With the above-mentioned sewage flow
rates, this allowed 6 hours of continuous flow.
To determine biological purification, 3 replicates of stand-
ard nutrient samples were taken for NH3, NO2, NO3, and PO* for
each sample. Samples of the effluent solution were taken on the
day of initial setup (March 16, 1979), and just before and after
the trickle feeding experiment. In addition, nutrient samples
were taken from the outflow of each of the four sets of test
troughs. These were taken just prior to the start of trickle
feeding; after two hours of feeding; after four hours of feeding;
when the feeding was stopped; and two hours after finishing the
trickle-feeding process. All nutrient samples were processed
and analyzed in the same manner as in section I of this volume.
RESULTS
In Part a of the flowthrough studies, Meraenaria and
Fundulus growth in pre-DAF cannery waste was investigated. The
Meraenavia data are represented by shell length and total weight
differences which resulted from six weeks of being fed pre-DAF
waste water. These data have been analyzed and are presented
in the graphs, Figures 10 and 11. Data recorded from the Fundulus
was also analyzed and are presented in the graph, Figure 12-
Results of Part b are evaluated on the basis of ammonia
levels remaining in the effluent. The data are oresented in
Figure 6.
DISCUSSION AND CONCLUSIONS
a. Simulated Ecosystem Growth
In this test pre-DAF cannery waste was trickle-fed to a
simulated ecosystem for a period of six weeks. The growth of
the organisms within this ecosystem was evaluated by measuring
and weighing two indicator species, Fundulus parvipinnis and
Meraenaria meraenavia, before and after this flow-through test.
-------
512
V D12
The results of the Mer?enaria measurements (Figures 10, 11)
indicate the extremely slow growth of this organism. No sig-
nificant growth occurred in weight or shell length during the
six week experiment. Mortalities were negligible in both
control and test clams, however, indicating that neither
treatment was detrimental to these organisms.
There were three test groups in the Fundulus experiment;
unfed controls, the treatment group fed pre-DAF waste, and
a second control fed the normal laboratory diet of trout chow.
As can be seen from Figure 12, the unfed control shows a definite
decrease in size over the experimental period. The pre-DAF
treatment group, however, did not significantly shrink and the
graph {Figure 12) indicates a slight increase in mean fish size.
From this one can conclude that trickle feeding of pre-DAF
waste is more beneficial to these organisms than not being fed
at all.
The control group fed trout chow seems also to show a
positive growth trend. Even though the growth in this group
is not significantly different from the treatment group, the
upward shift of the mean size of fish is greater than the
corresponding shift for the pre-DAF-fed fish. This is not sur-
prising, as these fish would be expected to show rapid growth
on this high protein balanced diet which they were accustomed
to eating.
In conclusion, the results of Part a suggest that the
cannery waste now subjected to TITP secondary waste treatment
could have a positive biostimulatory effect on some marine
species. At present, this nutrient is eliminated from the
receiving waters.
b. Biological Purification of TITP Effluent
The design of this experiment was intended to assess the
ability of a simulated ecosystem to purify TITP effluent
biologically. The effluent was initially diluted to a concen-
tration of 32% with sea water, inoculated with wild phytoplankton
from the Los Angeles Harbor, and incubated for twelve days to
produce the stock solution. This solution was then trickle-fed
at various concentrations to a simulated mussel-bed ecosystem.
Nutrient samples were taken throughout the experiment to
determine the biological purification ability of this system.
Only the ammonia values are discussed in the report.
As can be seen from the average ammonia values presented
in Table 6 , the initial 32 percent TITP effluent was very high
in ammonia. A value of 84.482 yg at/1 was found. After the
twelve-day incubation period, however, the major portion of the
ammonia was removed from the effluent solution. This was
probably due to a combination of uptake and evaporation. The
stock value prior to the start of trickle feeding (S0 in Table)
-------
V D13
513
was only 4 .622 ygat/L. This implies a high efficiency of ammonia
removal by the phytoplankton in the stock TITP effluent solution.
The low ammonia value in the effluent solution was reduced even
more by the end of the test, to 1.881 ugat/L (Sc in Table 6).
The ammonia removal efficiencies are within the range suggested
by Goldman and Ryther (1975) for mass cultures of marine algae.
This result implies that the algae by themselves are highly
efficient in purifying TITP waste of ammonia.
Since the stock solution ammonia levels are so much lower
than for normal harbor sea water (1.8-4.6 mg/L for the former,
as opposed to 10.8-11.2 jag at/1 in harbor water controls), the
only effect that was observed in the final outflows from the
troughs was an "ammonia reduction" in the higher concentration
of stock TITP solution. The resolution of ammonia analysis was
too low to detect significant ammonia removal by the simulated
mussel ecosystem.
In summary, the pre-DAF cannery wastes furnished a nutrient
source that could be distinguished as beneficial in Fundulus
tests. Meraenaria tests were not judged suitable for short-term
tests, due to very slow growth rates.
In flow-through, simulated ecosystem tests, wild phyto-
plankton cultures reduced ammonia levels greatly. Ammonia
levels were further reduced in the flow-through so that the
final values were below ambient seawater levels. The polyculture
treatment on a small scale suggests optional treatment modes as
well as natural biological processes in the harbor.
LITERATURE CITED See Section VIA
-------
SCAlf IN MILES
WILMINGTON
LONG
BEACH
TITP
A7
SAN
PEDRO
'A3
A3
CABftlLlO
BEACH/*
Al
Figure 1. Location of Mussel Racks for Biostimulation iri situ Studies.
-------
V D15
515
Depth
(meters)
0
Buoy
L
¦float
¦ net strivs
'ables
snaz> shackle
cables
Figure 2 . Construction Details for in situ Mytilus
Bioenhanceruent Experiment
-------
FIGURE 3. ABSOLUTE GROUP OF MYTILUS EDULIS
1 . 2
30 DAY GROWTH OF MYTILUS
	1	1	
u.
ON
J
6
u
3
0
L
O)
0.8'
0.6'
0.4"
0 . 2
~
~
o
Da
<
O
(-•
a\
2.5
3.5
4
i n I t I a I
4 5
ength (cm)
5.5
6.5
The regression I i no and 95% conf i dene# limits are plot lad with th« or i g i onaI data.
-------
V D17
517
TABLE 1 .	LINEAR REGRESSION ANALYSIS
30 DAY GROWTH OF MYTILUS, ABSOLUTE GROWTH
X represents Initial length (cm}.
Y represents growth Ccro}.
For 96 points supp I i ad, tha maan of X Is 3.8076.,
and tha moan of Y Is 0.4557.
Tha varIanca of X Is 0.6326 and of Y Is 0.0562.
Tho regression aquation Is- Y= -0.0539 X + 0.6610.
95 . 0X conf I denca I i m I ts for tha s I opa are —0 . 1 1 -40 & 0 . 006 1 .
ANALYSIS OF VARIANCE:
SOURCE SS
groups	5.3402
I i naar	0.!747
dev.	5.1655
error	0.0000
total	5.3402
df	MS
95	0.0562
1 0.1747
94 0.0550
0	0.0000
95
Fs	P
3.1794 0.078
-------
FIGURE h. PERCENT OF BODY LENGTH GROWTH
30 DAY GROWTH OF MYTILUS
Ln
(—1
00
<
O
b-1
00
6.5
Tho regression I i ri« arid 95% conf I denc« limits are plotted with the or I g i ona I data.
-------
V D19
519
TABLE 2.
LINEAR REGRESSION ANALYSIS
30 DAY GROWTH OF MYTILUS, PERCENT OF BODY LENGTH
X represents Initial length Ccm}.
Y represents X growth Ccm).
For 96 points supp Iiad, the mean of X Is 3.8076,
and the moan of Y Is I2.6748.
Tha varlanca of X Is 0.6326 and of Y Is 52.7906.
The regression equation Is- Y= -4.3020 X i- 29.0551.
95.2% confidence limits for the slope are -5.9524 & -2.6516.
ANALYSIS OF VARIANCE¦
SOURCE	SS	df
groups	5015.1113	95
Ii near	1112.1590	1
dev.	3902.9524	94
error	0.0000	0
total	5015.1113	95
MS
52.7906
1112.1590
41.5208
0.0000
26.7856
P
0.000
-------
520	V D20
TABLE 3.	HARTLEY ANOVA
IN SITU MYTILUS BIOENHANCEMENT
Factor*i
AC 173 m repl icatac
BC33 - depth
CC45 ™ location
DC4} m season
source of
sums of
degrees of
moon
var1 at 1 on
squares
f readout
squares
A
0.9788
16
0.0512
B
0.1929
2
0.0964
AB
2.3700
32
0.0741
C
1.6322
3
0.5441
AC
2.9020
48
0.0605
BC
0.7509
6
0.1251
ABC
S.9547
96
0.0620
D
I.9132
3
0.6377
AD
2.7129
48
0.0565
BD
0.5021
6
0.0837
ABD
7.6778
96
0.0800
CD
1.5481
9
0.1720
ACD
9.5763
144
0.0665
BCD
1.7497
18
0.0972
ABCD
18.5400
288
0.0644
-------
FIGURE 5c MYTILUS EDULIS GROWTH WITH
LOCATION AS A GROWTH FACTOR
A2
A3
A7
TITP boiI
-0. I

N-41S

N-413

N-459
N="474
-I	1	I	L.
_1	I	I	l_
-I	I	1	L.
0.1 0.2 0 3 0.4 0.5 0.6 0 7 0.8 0.9
Growth (cm)
1.1 1.2 1.3 1.4 1.5
* Th® ronga & maan + 2 S.E. or« shown.
-------
D»c-Jan
Oct—Nov
Aug—Sept
MAY-JUNE
FIGURE 6. MYTILUS EDULIS GROWTH WITH
SEASON AS A GROWTH FACTOR
-i	r

N-442
¦4*
N-460
N-461

N=398
-I	L.
-1	L.
u\
K)
NJ
O
N)
fvj
-0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 09 I I.I 1.2 1.3 1.4 1.5
Growth Ccm)
* Tha rang« & mman + 2 S.E. are shown.
-------
3 meters
2 meters
-i	r
FIGURE 7. MYTILUS EDULIS GROWTH WITH
DEPTH AS A GROWTH FACTOR

N-612

N-572

N=577
_1_

_L
-i	r
D
fo
OJ
-0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 t.2 13 14 15
Growth Ccm)	en
KJ
* Tha ranga & moan + 2 S.E. are shown.	to
-------
524	V D24
TABLE 4.
IN SITU MYTILUS BIQENHANCEMENT
ANQVA TABLE - 3 FACTORS WITH REPLICATES.
SOURCE OF	SUMS OF	DF MEAN Fg	P
VARIATION	SQUARES	SQUARES
SEASON	1.9132	3 0.6377 9.6577	<<.001 **
LOCATION	1.6322	3 0.5441 8.2402	<<.001 **
DEPTH	0.1929	2 0.0964 1.4599	>.20
LOCATION
& SEASON	1.5481	9 0.1720 2.6048	<.006 *
DEPTH &
SEASON	0.5021	6 0.0837 1.2676	>.25
DEPTH &
LOCATION	0.7509	6 0.1251 1.8946	>.05
ALL 3	1.7497	18 0.0972 1.4721	>„05
WITHIN GROUPS
(ERROR) 50.7125	768	0.6603
CONCLUSION t
SEASON AND LOCATION AND THEIR INTERACTION
ARE THE ONLY SIGNIFICANT (P<.05) FACTORS.
-------
FIGURE 8 *
0 . 6
e
o
X
2
0
L
CD

-------
PERCENTAGE BIOMA'iii INCRJiA:ii£ OF SETTLING ORGANISMS .	ui
co
A2
TITF Outfall.
<
A3
,50%
1500
500
1000
Distance From TIT? Outfall in Meters
F 1 GUREi 9.
i ercentage biomass increase of settling organisms in the proximity c.f the
TITF outfall during the one month period from July 14 - Aug 14, 19?'°.
-------
TABLE 5 , LiSTTLING HACK BIGL'NHANCH'Mb'NT STUDY DATA
:ett,l ing
Rack #
Weight
Initial
Location
let Month
Weight
1st. Month
BI omasa
1st Month
Location
2nd Month
Weight
2nd Month
Biomass
2nd Month
% Increase
of Biomass
17 1/4 °z
18 3/4 oz
1"< oz
18 t
A2
A 2
A2
28 oz
30 oz
2? 1/2 oz
2? oz
10 3/4 oz
TITP boil
11 1/4 oz
9 1/2 oz
8 1/2
oz
A7
A3
A2
42 oz
46 oz
36 1/2
OZ
24 3/4 oz
2? 1/4 oz
lft l/2 oz
42 oz
23 1/2 oz
13Q£
142%
95%
\77%
-------
Growth In M. marcanar i a
con tra
treatment
-0 . 3
FIGURE 10
N=99
N-99

0.2 0.3 0.4 0.5
length increase (mm)
LH
ro
CO
<
a
to
00
* The di ff«rence Is; not significant Cby anova, P"* 0.722); Rang® & mean + 2 S.E. are shown.
Homogens i ty o*T var I ances requ i remeni rea 1 i zed w I thocit transf or mat i on . CC=0 . 57 15).
-------
reatment
Growth in M. m e r
N=99
N-99
FIGURE
0.8 t
we j gh t Incr
2 . 6
The difference is not significant Cby anova, P= 0.3-40!); Range & mean + 2 S.E. are shown
Homogenei ty of vor i an cos requ i r emen t real i zed w i thou t t r ansf or ma t ion. CC=0.S027!>.
-------
Ui
u>
o
FunduI us Growth 	 			
unfed control, initial
unfad control, final
fed control, initial
fed control,, f I na I
<
a
U)
o
treatment, initial
treatmant, fInaI
FIGURE 12.
* Th® difference Is significant Cby anova, P"" 0.022}; Range & mean + 2 S.E. are shown,
liomogena i ty of var i ancas requ i remen t rea I i zed w i thou t tr ansformat i on . CC=0 . 29 17).
i	1	r
_1	1	,	1	,	1	,	r
, N-10
4=3-
N=6
I	E==
N=I0
N=8
E3
-HZ
N-1 0
N-8
3 —I
-J	I	L.
I 	1	I 		I	L	l_	J	l_
0
2 3 4
5 6 7
wo i gh t C gins . )
8 9 10 11 12
-------
V D31
533.
TABLE 6 . AVERAGE AMMONIA VALUES FOR THE FLOW-THROUGH
BIOENHANCEMENT STUDY (VALUES IN jjg AT NH 3 /L)
ELAPSEP TIHS
CONC. OF
2° EFFLUENT	INITIAL	2 HRS	5 HRS	6 HRS	FINAL
CONTROL
(0%) 10.8825 10.533	11.178 10.748 11.205
1.35% 10.318	10.399 9.996 11.125
5.68% 10.452 10.184	10.318 9.861 9.512
10% 10.238 7.980	8.249 9.861 12.360
STOCK SEWAGE SOLUTION AMMONIA VALUES
INITIAL TI TP	84.482
SO	4.622
SX	1.881
AMBIENT HAROR WATER	10.8-11.2
ALL VALUES GIVEN ARE THE AVERAGE OF THREE REPLICATE SAMPLES.
-------
Intentionally Blank Page
-------
VIA
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547
EAP
ECOLOGICAL ANALYSIS PACKAGE
METHODS SECTION
****** DISCRIMINANT ANALYSIS ******
PAPER #1
WRITTEN BY
ROBERT W. SMITH
DEPARTMENT OF BIOLOGICAL SCIENCES
UNIVERSITY OF SOUTHERN CALIFORNIA
LOS ANGELES, CALIFORNIA 90007
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548
INTRODUCTION
There are tuo basic approaches in discriminant analysis. Both
involve a priori definition of tuo or more groups of observations. The
most common use of discriminant analysis involves assigning unknown
observations to one of the defined groups (Lachenbruch, 1975;
Gnanadesikan,1977). The second approach involves the description and
testing of between-group differences (Hope, 1969; Cooley and Lohnes,
1971; Green, 1976; Pimentel and Frey, 1978). The latter approach is
discussed here.
Suite often in ecological-survey work, one of the goals is to
study the relationships between the biological and environmental
patterns. As will be shown, discriminant analysis is well suited for
this purpose.
The general idea of discriminant analysis is illustrated with
an example. Fig 1A shows a dendrogram defining two groups of sampling
sites. It is assumed that this cluster analysis is based on the biotic
data collected at the sites . This would be one way to summarise the
biological patterns in the study area.
Let's say tuo environmental variables	(salinity and depth) are
also measured at each site. Fig 1B shows	what might result if the
sites were plotted according their level of	salinity and depth. Note
the following.
1)	All sites in dendrogram group 1	(sites A-E) are found in
shallow depths.
2)	All sites in group 2 (sites F-L) are found in deeper depths.
3)	The salinity values found at the sites in the groups are
broadly overlapping.
With this type of result, displayed in this manner, it is
evident that the biological pattern may somehow be related to
variations in depth, and probably not related to the level of salinity
found at the sites.
Fig 1C illustrates a more complex hypothetical result. Again,
the sites are plotted according to the depth and salinity values.
However, the values of both variables are broadly overlapping, i.e.,
sites in both groups are found more or less at all measured values of
depth and salinity. In spite of this, the group members are completely
separated in this plot, indicating that these two variables may
somehow be related to the group separation.
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Figure 1. Hypothetical survey data used to explain the idea of
discriminant analysis.
A. hypothetical dendrogram
based on biotic data
sites	>	ABCDEFGHIJK
I	I I	I
groups	>	1	2
environmental parameters at the sites
36.0-	C. 36.0	B
IK	I	L
I	G	*
I C	*
depth (m)	(asterisks) *	depth
*

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550
If all points in the plot (fig 1C) were perpendicularly
projected onto "line A", the point projections for the two groups
would be completely separated. In effect, a new variable which
separates the groups has been defined. The values of this variable are
the values of the projections onto the diagonal line. Projections onto
the line will be correlated with the values of both salinity and
depth. This new variable could be thought of as a "salinity-depth
considered simultaneously"-type parameter. The conclusion to be drawn
from fig 1C would be that the group separation (biotic pattern) could
be related to both salinity and depth, but to account for the result,
both variables must be considered simultaneously.
In fig 1B, note that if a "new variable" which would best
separate the groups were to be defined in the same manner as in fig
tC, the position of the line representing the variable would lie
parallel or nearly parallel to the depth dimension. Thus, the new
variable would essentially be a depth variable, with little, if any,
component of salinity.
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551
THE METHOD OF DISCRIMINANT ANALYSIS IN GENERAL TERMS
Discriminant analysis attempts to find these "new variables"
which will best separate the predefined groups. In general terms, the
process can be summarized as follows.
1)	A priori groups are defined according to a biological
criterion.
2)	A hypotetical, multidimensional "space" is set up. The
dimensions of this space represent the measured environmental
variables. The position of a site (sample, observation, etc.) will
depend on the level of each variable measured at the site.
3)	A new variable, which best separates the groups, is defined.
This variable is represented in the space by a line called a
discriminant axis (e.g., "line A" in fig 1C). The value of this new
variable at a site is the perpendicular projection of the site point
onto the discriminant axis (see fig 2). The value of the projection is
called a discriminant score.
*4) The position of the discriminant axis in this space will
depend on which combination of variables best separates the groups.
The discriminant axis will not extend far into dimensions which
represent variables showing little relationship to group separation
(e.g., salinity in fig 1B). The discriminant axis will be situated
mostly in dimensions representing variables which are related to group
separation (e.g., depth in fig IB, or both depth and salinity in fig
1C) .
5) When more than two groups have been defined, more than one
discriminant axis may be required to separate the groups. Fig 3
illustrates this concept. Note that the first discriminant axis
separates group Y from groups X and Z, while the second axis separates
group Z from groups X and Y. To avoid redundancy of information on the
different axes, the site scores on the different axes are made to be
uncorrelated. The axes are not necessarily at right angles to each
other (Green, 1976). The axes are usually ordered according to the
amount of group separation accounted for, i.e., the first axis will
show the most group separation, the second axis, the second most, and
so on.
-------
552
Figure 2. An illustration of the idea of scores as perpendicular
projections of points onto the discriminant axis.
15
variable A
I
10
/	I .
-10 *	I .
*
*	5	Y
/ . I
-5 * . I
projection of	>* I
point X (score	* I
of point X = -2.5)	I	I	._!	I
0 *
* 5 . 10	15
*
/ .	variable B
projection of point Y	> *
(score of point Y = 6)	*
*
/
+ 10 *
x
discriminant axis	> *
(asterisks)
-------
553
Figure 3. Discriminant analysis with three groups (X,Y,Z). Notice
that tuo discriminant axes are required to separate the
three groups.
I	Z
variable A	I	Z
I	Z Z
I	Z
*	I	X Z	<	second
*	I	XX.	discriminant
*	I X X	.	axis (dots)
*	1
* 1	X .
* I
* !	Y
first 	> * !	Y Y
discriminant axis *! . Y Y Y
(asterisks)	1	
*
*
*
*	variable E
-------
554
DISCRIMINANT COEFFICIENTS
Discriminant coefficients are used to indicate which original
variables are related to each axis. Each axis has a separate set of
coefficients, with one coefficient for each original variable. The
magnitude of the absolute value of a coefficient is relative to the
importance of the corresponding variable on the axis in question. For
example* if the data in fig 1B were analyzed, the coefficients uould
appear as
axis 1
salinity	0 . 7
depth	99.3
The coefficients from the data in fig 1C uould appear as
axis 1
salinity	36 . 2
depth	63.8
These results agree with the observations made above, mainly
that depth was mostly related to group separation in fig 1B, and both
variables were related to group separation in fig 1C.
The coefficients are adjusted to account for the differing
scales of the original variable. There are three methods by which this
is accomplished. One is to standardize the coefficeints for a variable
by the total standard deviation for that variable (Cooley, and Lohnes,
1971). The second is to standardize the coefficients by the
within-group standard deviation of the the corresponding variables
(Green, 1976). The third technique involves the the computation of the
coefficients of separate determination (Hope, 1969). These
coefficients are already adjusted for scale, and no standardization is
required. The coefficients given in the above examples are
coefficients of separate determination.
-------
555
MULTIVARIATE VS. UNIVARIATE METHODOLOGY
The example used in fig 1C illustrates the importance of using
a multivariate technique in such cases. A multivariate technique
considers all the variables simultaneously instead of one at a time as
in univariate analysis.
To illustrate the increased power of the multivariate method,
univariate F tests (one-way ANOVA) were run to try to detect group
differences in each of the two variables (data from fig 1C). Neither F
value was significant at the 5% level. In contrast, the
discriminant-analysis test for group differences (see note 9, fig 4)
was highly significant (P << .001). The latter result, of course, is
the desired one, since there are group differences in relation to the
variables.
DISCRIMINANT ANALYSIS CALCULATIONS
No attempt is made here to completely explain the discriminant
analysis calculations. The reader should consult the multivariate
texts mentioned above (especially, Green, 1976) for more details. Fig
4 summarizes the calculations. Matrix notation is used. There are v
variables and n observations (sites, etc.). Sample calculations are
shown in Appendix A.
-------
556
Figure	Flow chart of the discriminant analysis calculations. See
accompanying notes for additional details.
total
note 1
data
matrix
note 2
uithxn
between
I
I note 3
!
< —
equations
solved for
A , L
k k
(note 4)

eigenvalue
for axis k
eigenvector for
axis k
coefs. of
sep. det.
	>
note 6
note 5
scores
note 7 I
1
std. coefs .
(by total SD)
note 8
std. coefs.
(by within SD)
-------
557
Notes fdr Figure 4.
********** note 1 **********
The data are centered by the overall variable mean.
If Z is the centered data matrix, then
with
3 = x - x
kj kj j ,
n
suric x )
i=i ij
n
The T matrix is calculated from Z as follows
T = Z' Z
The element in the kth row and the jth column of T would be
t = SUM ( z s )
kj i=1 ik ij
or. in terms of X,
t = SUM ( (x - x )(x - x ) )
kj i=1	ik k ij j
Matrix T is symmetrical.
-------
558
Notes for Figure 4.
4k »|i	^	^ | q ^ ^ ^  #|C If! * )|t )K
The data for each predefined group are worked on separately.
Define Y as a (m x v) data matrix containing the observations in
h
group h. The data are centered by the variable means for the group
If C is the centered matrix for group h, then
h
c = y - y
kjh kjh j h
The calculation of the w matrix for group h is as follows.
W = C' C
h h h
The element in the kth row and the jth column of W is
h
m
h
w = SUM ( c c )
kjh i=1 ikh ijh
or in terms of Y,
m
h
w = SUM ( (y -y )(y -y ))
kjh i=1	ikh ih ijh jh
( note 2 continued on next page )
-------
559
Notes for Figure 4
( note 2, continued )
To obtain the final W matrix, the W matrices for each group
h
are summed, i.e.,
W = W + W + 	 «
1 2	g
where g = the number of groups. This pooled matrix summarizes the
within-group variation and covariation.
Matrix W is symmetrical.

The simplest way to obtain matrix B is as follows
B = T - W
The element in the kth row and the jth column of B is equivalent to
S	_ _ _
b = SUM CnCx - x )(x -x)) ,
kj h=1 h kh k jh j
where g is the number of groups, n is the number of observations in
h
group h, x is the mean of variable k in group h, x is the mean of
kh	jh
variable j in group h, and x is the over-all mean of variable k, and
k
and x is the overall mean of variable j. This matrix summarises the
j
variation and covariation of the group means.
-------
560
Notes for Figure 4.
************** note 4 ***************
The eigenvalues and eigenvectors of the asymmetric
-1
matrix U B are found. The solutions for these equations will have
the following property.
A * B	A	I
k	k	I
k A' W	A	I
k	k	i MAX
where L is the eigenvalue for axis k, and A is the eigenvector for
k	k
axis k. In words, this means that the eigenvalue of axis k is equal
to the maximized ratio of 1) the between-group sum of squares of the
discriminant scores, and 2) the within-group sum of squares of the
discriminant scores for axis k. This maximization will emphasize
variables which contribute a relatively large amount of between-group
variability relative to the within-group variability.
This maximization is constrained in that
i.e., each eigenvector must be of unit length. This avoids a solution
which makes A' B A (or A' W A ) indefinitely large (or small) by
k k	k k
making the entries of A arbitrarily large (or small). A derivation
k
of these equations is found in Green (1976; 247-254).
-------
561
Notes for Figure 4.

The scores on axis k (S ) are calculated as follows.
k
S
X A
k
k
S will be the kth column, in matrix S, which contains the scores for
k
all p axes. The scores for each axis can be standardized to unit
variance by dividing the eigenvector elements by the overall standard
deviation of the corresponding variable, i.e.,
where a is the eigenvector element for variable i on axis k, and
1/2
p = a / q
ik ik k
ik
1
q = A' C 	
k n-1
T ) A
k
CCooley and Lohnes, 1971; 31,217).
P
k
would be used in subsequent calculations instead of A .
k
k
-------
562
Notes for Figure 4.
************** note 6 ***************
The coefficients of separate determination for axis k are
calculated as follows:
D = Z T Z U	(Hope, 1969)
k k k
where Z is a diagonal matrix with the elements of A in the principa
k	k
diagonal, and zeros elsewhere. The matrix U is a (v x 1) column
vector of ones. D is the kth column of matrix D (D contains the
k
coefficients for all p axes, with axes in the columns).
Theoretically, all the coefficients should be positive. This,
however, is not always the case. Experience has shown that the
magnitude of the absolute value corresponds to the importance of the
variable. The coefficients can be expressed as percents of the
total of the coefficients for the axis (only the absolute values used
*************** note 7 *****************
These coefficients are calculated as follows
1/2
e = a (t / (n-1) )
jk jk jj
where e is the standardised coefficient for variable j on axis k,
jk
a is the eigenvector element for variable j on axis k, t is the
jk	j j
jth diagonal element in matrix T (the centered, overall sum of square:
of variable j), and n is the total number of observations. The seconc
term in the product standardizes a to make all coefficients
jk
comparable (the variables will usually be measured on different
scales). The coefficients can be expressed as the percent of the tota:
of the absolute values of the coefficients for an axis.
-------
563
Notes for Figure 4.

These coefficients are calculated as follows=
1/2
f = a (u /(n-g) )
jk 3k jj
where f is the standardized coefficient for variable j on axis k,
jk
a is the eigenvector element for variable j on axis k, u is the
jk	j j
jth diagonal element in matrix W (the centered, within-group sum
of squares of variable j), n is the total number of observations,
and g the number of groups. The second term in the product
standardizes a to make all coefficients comparable (the variables
jk
will usually be measured on different scales). The coefficients
can be expressed as the percent of the total of the absolute values
of the coefficients for an axis.
£ <1* *4* IK	X ^ q ^ ^ ^ It* Itl 24*	Ifl HC HC <11 H!
The significance of group separation on axis k can be tested
by calculating
chi square = ( n-1-1/2(v+g) ) ln(1+L )
k
with
D.F. = v + g - 2 k	(Hope, 1969; 118)
here n = # observations, v = # variables, g = # groups, L = eigenvalue
k
for axis k, and In = a natural log operation. The assumptions of
the test are summarized in Green (1971). In addition, the groups
must be non-overlapping (Green, 1976; 278). In the author's exper-
ience, the assumptions are rarely met with ecological-survey data, but
under certain conditions the test may be fairly robust (see pp 33-34).
Observation of score plots for the sampling sites will usually be
sufficient to determine whether the groups are well separated or not.

-------
564
THE SELECTIOK OF GROUPS PRIOR TO THE DISCRIMINANT ANALYSIS
Groups can be chosen in any way relevant to the analyst. On
such method has been mentioned in the introduction, i.e.,
classification (cluster) analysis prior to the discriminant analysi
(Smith, 1976; Green, 1977; Bernstein et al, 1978). Smith (1976
142-145) discusses some aspects of group selection with hierarchica
classification. It is concluded that it may not always be too critica
at which specific level the groups are delimited.
An alternate technique for forming groups of observations is t
use the species data matrix directly (Green, 1971,1974; James, 1971
Dueser and Shugart, 1978). This technique is illustrated in fig 5
Here each group corresponds to a single species. The variables whic!
tend to correspond with species separation (in space, time, etc.) wil.
be emphasised in the discriminant analysis results. Note that a singli
observation (site) may be in more than one group. This violates one o
the assumptions of the chi-square test for
9 for fig 4).
results
. Note
group.
This v
group
separa
-------
565
Figure 5. The formation of groups directly from the biotic data
matrix.
A. data matrix
sites
12 3 15
A
species B
C
3 10 0 2
2 0 0 2 1
0 0 3 2 0
B. sites in groups representing each species, i.e., the site
in which each species occurs.
sites in group
A	1,2,5
B	1, 4, 5
C	3,4
-------
566
WEIGHTED DISCRIMINANT ANALYSIS
It will often be the case that group members mill vary in hou
representative they are of their own group. Weighted discriminant
analysis allows for weighting the calculations for a group to give
more.emphasis to the "better" members of the group in question (Smith,
1976). As will be shown, this technique can also be used to input
(into the calculations) information concerning between-group
biological similarities. This can significantly increase the power and
accuracy of the analysis. It will also be shown that this technique
can even be used without any a priori group definition.
Weighted discriminant analysis calculations.
The only changes in the calculations involve the sums of
squares and cross-product matrices. Both weighted and unweighted
calculations are included for contrast. Sample calculations are shown
in Appendix B.
W matrix. In fig M (note 2) it was shown that for regular
discriminant analysis, the element in the kth row and the jth column
of W (the contribution of group h to the pooled w matrix) was
h
w
k jh
m
h
sun
i= 1
( ( y - y )(y -y
ikh kh ijh jh
) )
The weighted calculations are
k jh
m
h
SUM
i= 1
((y - y' )(y -y' ) u
ikh kh ijh jh ih
-------
567
where
m
h
SUM ( y u )
_	i=1 ikh ih
y * =		( a weighted mean),
kh	m
h
SUM (u )
i= 1 ih
and u is a weight which is proportional to how well observation i
ih
fits in group h. This formula allows the observations more
representative of the group in question to receive greater weight
in the calculations for the group. This is done in two ways.
1) Since a weighted mean is used, the observations with higher weight
(i.e., more representative of the group 3 will have more influence on
the mean value, and 2) the cross product itself is weighted, thus
the observations more representative of the group will add more to the
sum of the cross products for the group.
T matrix. In fig 4 (note 1) the element in the kth row and jth
column of T was shown to be
n	_	_
t = SUM ( (x - x )(x - x ) )
k j i= 1	ik k i j j
The weighted calculations are as follows. The overall weighted mean
(to be used instead of x ) is
k
m
g h
SUM SUM ( x u )
_	h=ti=1 xkhih
k	m
g h
SUM SUM ( u )
h=1 i=1 ih

-------
568
where x is the ith observation of variable k in group h, and u is
ikh	ih
the weight of the ith observation in group h.
The element in the kth row and jth column of T is
m
3 h	_	_
t = SUM SUM ( ( x - x' )( x - x' ) u )
kj h=1 i=1	ikh k	ijh j ih
This is similar to the calculations for the U matrix, except the
weighted overall mean is used instead of the weighted group means
B matrix. The B matrix is (as with regular discriminant analysis)
B = T - W
With the usual discriminant analysis calculations, b is equivalent
kj
to
9	_ _	_ _
b = sun (n (x -x)(x -x))
k j h= 1 h kh k	j h j
In the weighted calculations, b is equivalent to
kj
9	_ _	_ _
b = SUn ( p ( y' - x' ) ( y' - x' ) )
kj h= 1 h kh k	jh j
-------
where
m
h
p = SUM ( u )
h i=1 ih
Here x' is the overall weighted mean for variable k (see above),
_ k •
and y' is the weighted mean for variable k in group h (see above),
kh
These formulae are similar to the non-weighted method, except that
weighted means are used instead of regular means, and the sum of
weights for the group is used instead of the number of observations
for the group.
Weighted discriminant analysis with groups directly from the
species-site data matrix.
Fig 6A depicts a hypothetical situation with two species (A and
10 potential sampling sites along an environmental gradient
E) presumed to be important in the separation (in an
ecological sense) of the two species.
If sites 3-8 were sampled, the data might appear as in fig 6B.
Normally, the species data would be used to select groups and variable
E would be used in the discriminant analysis. Note, however, that as
far as group membership is concerned, both groups (one group
corresponding to each species) would be identical, since both species
occur at all sites. With regular discriminant analysis, these two
species could not be differentiated (with respect to variable E),
since there would be no between-group variation (the means of variable
E for both groups would be the same, see fig 6C ).
On the other hand, if weighted calculations are used, the group
means can be differentiated as desired. This is illustrated in fig 6D,
where the species abundances are used as weights. These are
appropriate weights, since the more a species occurs at a site, the
more representative of the group (species) is that site. Normally,
some standardized measure (such as species-maximum standardised data)
of species importance should be used instead of raw abundance counts.
This will prevent the more abundant species from dominating the
analysis.
B
)
, and
(
V
ariable
e
c
ologica
-------
570
Figure 6. Illustration of the advantages of weighted discriminant
analysis when groups are chosen directly from the data.
A. two hypothetical species distributed along an environmental
gradient.
20 _ species A
I (dots)
I
10
* *
	>. * *
*
*	. *<	 species B
(asterisks)
* *
I
sites	12	3	4	5	6	7
E	>
B. data values
sites
3	4	5	6	7	8
species
A
. 12
20
22
17
1 1
4 .
species
B
6
14
20
22
17
12 .
variable
E
2
4
6
8
1 0
12 .
¦ I ¦
9
1 0
I <— used as
I	weights
C. group means for variable E
E = E = 42/6 = 7 (no group differentiation)
A B
D. weighted group means for variable E
12(2)+20(4)+22(6)+17(8)+11(10)+4(12)
E' = 	 = 530/86 = 6.16
A	12+20+22+17+11+4
6(2)+14(4)+20(6)+22(8)+17(10)+12(12)
E' = 	 = 678/9 1 = 7.45
B	6+14+20+22+17+12
-------
571
Even if the groups (species) are not completely overlapping,
the weighted method may often have advantages. With the weighted
calculations, data from all sites with different species importance
values are effectively used in trying to differentiate the groups,
whereas with the unweighted calculations, only the sites of
non-overlap can be used to differentiate the groups. Since more
information is used, the weighted method should often be more robust
and accurate.
Weighted discriminant analysis with groups defined from cluster
analysis.
Fig 7 shows a hypothetical dendrogram from a hierarchical
cluster analysis based on biotic data. From the dendrogram, it can be
seen that there are four important pieces of biological information
concerning the the groups . These are '•
1)	group membership, i.e., sites 8 9 10 are in group 1, etc.;
2)	strength of group membership, e.g., site 7 is the weakest
(most unlike the other members) member of group 3;
3)	"cohesiveness" of a group in general, e.g., the members of
group 1 are all more similar to each other than is the case with the
other groups; and
4)	inter-group reltionships. Note that groups 2 and 3 are more
closely related to each other than either one is to group 1.
With regular discriminant analysis, only the first (group
membership) is input. The other information can be very important. One
would want the analysis to emphasize environmental variables which
most closely follow all four patterns, not just the first. For
example, the three groups in fig 7 might be quite different in levels
of both salinity and depth, but groups 2 and 3 are closer in depth to
each other than to group 1, and groups 1 and 2 are closer in salinity
to each other than to group 3. A good analysis would indicate that
depth was more important than salinity (for group separation) since
the depth pattern more closely fits the biological pattern (see (4)
above). All four pieces of biological information can be input with
weighted discriminant analysis. A technique for doing this is shown
be low.
-------
572
Figure 7. Hypothetical dendrogram used to illustrate the advantages
of weighted discriminant analysis.
sites	> 89 10	1234567
I	i I	I I	I
groups	>	1	2	3
-------
573
The average group-similarity matrix (GRSIM).
The first step is to create a matrix which describes the
relationships between every group and every site. This will be called
the GRSIM matrix.
All the information r
matrix on which the cluster
distance matrix from whic
elements above the main
similarities (the distances
the GRSIM matrix is simp
(corresponding to a co
(corresponding to a row)
calculations, and fig 8B
similarities in fig 8A and g
equired is in the dis
analysis is often ba
h the dendrogram i
diagonal are distance
subtracted from a con
ly the average simila
lumns of the mat
in question. Fig
shows the complete
roup membership as in
tance (or similarity)
sed. Fig 8A shows the
n fig 7 was made. The
s and those below are
stant). An element of
rity between the site
rix) and the group
8C shows some sample
GRSIM matrix from the
fig 7.
k
-------
574
Figure 8. The calculation of the average group-similarity
matrix (GRSItt). Data fits the hypothetical dendrogram
shown in fig 7.
A. distance (upper-right triangular) and similarity (lower-left
triangular) matrices
sites
4	5
1 0
sites
1
2
3
4
5
6
7
8
9
1 0
4 . 0
2 1.0
2 1.0 19.4
4.0 10.8 14.7 14.0 20.2 22.0 23.6 24.5
5.6 10.1 14.2
14.2	14.9 17.8
10.3	10.8 14.0 21.0
4.0
9.7 18.5 18.2 19.5 20.5
7.2 11.0 10.0 17.0 21.1 23.0 23.3
5.9
4 . 0
11.0 15.3 15.0 19.1 21.0
4.8 6.5 8.0 15.0 17.4 13.6
10.0 17.3	19.8	19.1
7.6 19.0	21.1	20.0
11.4 22.5	24.9	24.0
15.0	17.3	15.1
2 . 5
3.0 6.8 3.9 7.7 6.0 2.5 10.0
1.4 5.5 2.0 5.2 3.9 0.1 7.7 22.5
0.5 4.5 1.7 5.9 5.0 1.0 9.9 22.4 22.0
2 . 6
3.0
( fig 8 continued on next page )
-------
575
( fig 8, continued )
B. the average group-similarity matrix (GRSItt)
1
sites
4	5	6
8
10
group 1 1.6 5.6 2.8 6.3 5.0 1.2 9.2 22.5 22.3 22.2
group 2 21.0 20.2 20.2 15.6 11.8 10.4 6.4 4.6 3.0 2.2
group 3 10.1 11.9 13.7 18.4 19.8 17.9 15.3 6.6 4.2 5.5
group 2
group 3
group 1
C. sample calculations for elements of GRSIM matrix
( s = similarity between sites i and j )
i j
site 2 - group 1
(s +s +s ) / 3 = (6.8+5.5+4.5)/3 = 16.8/3 = 5.6
2,8 2,9 2,10
site 3 - group 2
( s + s ) / 2 = (21+19.4)/2 = 20.2
3,1 3,2
k
-------
576
Application of weights.
The elements of the GRSIM matrix are now used as weights in a
weighted discriminant analysis. At this point, each site is considered
to be a potential member of each group, i.e., each site has a weight
indicating how well it fits into each group. In the calculations for
group- 1, the weights used for each site would be the corresponding
elements in the first row of the GRSIM matrix. For group 2, the
weights would be in the second row of the GRSIM matrix, and so on.
It can now be shown that all four pieces of biological
information about the groups are available to the analysis in the
GRSIM matrix of weights.
1) Group membership. Note that for each group (row of the GRSIM
matrix), the highest average-similarity values (i.e., weights) are for
the actual group members. For example, row 1 represents group 1, which
consists of sites 8, 9 and 10. Sites 8, 9, and 10 have the highest
weights of all the sites. This would be expected since they are the
group members. Thus, group membership is conveyed since the actual
group members should have the highest weights in the calculations for
the group in question.
2) Strength of group membership. Site 7 is the "weakest" of the
actual members of group 3. The average-similarity values (row 3, fig
8B) in the GRSIM matrix for the members of group 3 (sites 4-7) are
18.4, 19.8, 17.9, and 15.3. Note that the lowest value is 15.3, which
corresponds to site 7. Thus, of the actual members of group 3, site 7
will receive the lowest weight, which is consistent with its
biological relationship to the rest of the group members.
3) "Cohesiveness" of a group in general. Group 1 has the
highest internal biological similarity of the three groups (connected
lowest on the dendrogram). The average similarities of the members of
group 1 with their own group are 22.5, 22.3, and 22.2. The average
similarities of the members of group 2 with their own group are 2 1.0,
20.2, and 20.2; the average similarities of group 3 with its own
members are 18.4, 19.8, 17.9 and 15.3. Of the three groups, the
members of group 1 have the highest average similarity values with
their own group. This results in group 1 receiving more overall weight
per site than the other groups. This makes sense since group 1 is
closer to a real homogenous group than are the other groups. This can
be important since the analysis will try to minimize the within-group
variation in the discriminant space (along with the maximization of
the between-group variation). The lower weights for the "looser"
groups will prevent the analysis from emphasizing variables which will
minimize the distance (in the discriminant space) between sites which
are not really that biologically similar. This same arguement applies
-------
577
to (2) above.
4) Inter-group relationships. Groups 2 and 3 are biologically
more similar to each other than they are to group 1 (fig 7). This
information is also available in the GRSIM matrix. In the row for
group 2 Crow 2, GRSIM matrix), the sites that are in group 3 (sites
4-7) show higher average similarity values (15-6, 11.8, 10.4, and 6.4)
than do the sites in group 1 (4.6, 3.0, and 2.2). Also, in rou 3, the
values for group 2 (10.1, 11.9, 13.7) are higher than those for group
1 group 1 (6.6, 4.2, 5.5). This indicates that in the calculations for
group 2, the sites in group 3 will get more weight than the sites in
group 1, and also that the group 3 calculations will contain
relatively higher weights for group 2. Thus, it can be seen that the
more similar groups will have higher average similarities for each
other's members.
It should be noted that this GRSIM approach is only one of many
possibile techniques to obtain weights for the sites. All that is
required is a GRSIM-type matrix which expresses the relationships
between the defined groups and each site. For example, probabilities
of group membership could be used instead of average similarities.
-------
578
Weighted discriminant analysis with no groups defined.
One way to avoid defining groups at all is to consider each
site a group by itself. When this is done, the GRSIM matrix will
simply be the inter-site similarity matrix. Weighted discriminant
calculations are then carried out with the GRSIM matrix as weights in
the manner described above.
This is a good way to directly analyze the distance (or
similarity) matrix (i.e., the biological patterns) without any prior
clustering procedure. This has the advantage of saving the clustering
computation time and avoiding any errors (of group membership) that
the clustering technique may introduce. (However, the weighted
discriminant calculations themselves will be longer, since more groups
will be involved). Such a technique also eliminates the burden of
deciding where and how to define group membership from the clustering
results.
The fact that no group membership need be defined suggests that
weighted discriminant analysis could in some cases be a replacement
for a canonical correlation analysis, which is used to study the
relationships between two sets of variables. Gauch and Wentworth
(1976) demonstrate how the strict assumptions of linearity make
canonical correlations unsuitable for some types of ecological data.
Weighted discriminant analysis only requires that the variables used
in the calculation of the sum of squares and cross-products matrices
(W, B, T) linearly separate the groups (observations in this case).
This assumption can easily be met with most kinds of
biotic-environmental data sets. When some variables separate the
observations in a monotonic but non-linear fashion, a transformation
of the corresponding variable(s) will usually make the relationships
more linear.
-------
579
DISCRIMINANT ANALYSIS WITH SPECIES AS VARIABLES
When groups
defined (usually by
species importance
use) to=
of biologically similar sampling sites have been
cluster anaysis), discriminant analysis with the
values as variables has been used (or tested for
1 ) determine which species were mainly involved in causing the
group separations (Cassia, 1972; Gringal and Ohmann, 1975);
2)	observe the relatinships between the groups in the
discriminant score plots (Norris and Barkham, 1970; Cassie, 1972;
Gringal and Ohmann, 1975; Holland and Polgar, 1976; Holland et al,
1977);
3)	use the results as an indirect ordination technique in the
same manner that principal components, reciprocal averaging, polar
ordination, multidimensional scaling, etc. are used with biological
data (Kessell and Whittaker, 1976); and
4) test the significance of the group separation
Here also, the regular discriminant analysis only considers
group membership, while the weighted version can use additional intra-
and inter-group biological information in the calculations. This
additional information input should give better results in most cases
when (1), (2), or (3) above, is the goal of the anaysis.
As far as testing the significance of group separation is
concerned, weighted discriminant analysis is presently of little help
since the groups are, in effect, overlapping (this violates an
assumption of the tests). Even without overlap, it is doubtful that
the significance tests would be completely valid due to the usual
nature of species abundance data. For example, one of the assumptions
of the method (when statistical tests are applied) is that the
withm-group dispersion matrices (W matrix divided by D.F.) are
statistically equal (Green, 1971). If the survey in question covers
more than a single homogeneous habitat, then one would expect some
species to occur in some groups but not in others. In the groups where
such a species occurs, the dispersions would be some value other than
zero; but in groups where the species is absent, the dispersions would
be zero. This alone would lead to quite different within-group
dispersion matrices for the various groups. In the experience of the
author, whether species or environmental data are used in the
discriminant analysis, this assumption is almost always violated, but
the extent of the violation is greater with species data.
On the other hand, discriminant analysis can be fairly robust
even when the within-group dispersion matrices are not statistically
-------
580
equal (Cooley and Lohnes, 1971; Pimentel and Frey, 1978). The method
becomes more robust as the group sample sizes become larger and more
equal (Ito and Schull, 1964). It is not known how the robustness is
affected by truncated variables (e.g., some species-count data).
If the groups are defined using the species data (as would be
the case in a cluster analysis), and then these groups are used in a
discriminant analysis with species as variables, the usual probability
tests are invalid even if all the other assumptions are met. This is
because the statistical tests assume that the groups were defined by a
planned, .a priori procedure (Sokal and Rholf, 1969; 226-227). When the
data are used to suggest the groups, this assumption is obviously
violated.
-------
581
REFERENCES
Bernstein, B.B., R.R. Hessler, R. Smith, P.A. Jumars, 1978. Spatial
dispersion of benthic Foraminifera in the abyssal central North
Pacific. Limnol. Oceanogr. 23(3): '401-416.
Cassie, R.M., 1972. Fauna and sediment of an intertidal mud-flat: an
alternate multivariate analysis. J. Exp. Mar. Biol. Ecol. 9:
55-64.
Cooley, W.W. and P.R. Lohnes, 1971. Multivariate Data Analysis. John
Wiley and Sons, New York: 364 pp.
Dueser, R.D. and H.H. Shugart Jr. 1978. Microhabitats in a
forest-floor small mammal fauna. Ecology 59(1): 89-98.
Gauch, H.G. and T.R. Wentworth, 1976. Canonical correlation analysis
as an ordination technique. Vegetatio 33(1): 17-22.
Gnanadesikan, R., 1977. Methods For Statistical Data Analysis Of
Multivariate Observations. John Wiley and Sons, New York: 311 pp.
Green, P.E., 1976. Mathematical Tools For Applied Multivariate
Analysis. Academic Press, San Francisco: 376 pp.
Green, R.H., 1971. A multivariate statistical approach to the
Hutchinsonian niche: bivalve molluscs of central Canada. Ecology
52: 543-556.
Green, R.H., 1974. Multivariate niche analysis with temporally varying
environmental factors. Ecology 55: 73-83.
Green, R.H., 1977. Some methods for hypothesis testing and analysis
with biological monitoring data. Biological Monitoring Of Water
And Effluent fiuality. ASTM STP 607, John Cairnes, Jr., K.L.
Dickson, and G.F. Westlake, eds . American Society for Testing and
Materials: 200-211.
Gringal, D.F. and L.F. Ohmann, 1975. Classification, description and
dynamics of upland plant communities within a Minnesota
wilderness area. Ecological Monographs 45: 389-407.
Holland, A.F., H.K. Mountford, and J.A. Mirhursky, 1977. Temporal
variation in upper bay mesohaline benthic communities. I. the 9-m
mud habitat. Chesapeake Science, 18(4): 370-378.
Holland, A.F. and T.T. Polgar, 1976. Seasonal changes in the structure
of an intertidal community. Marine Biology 37: 341-348.
k.
-------
582
Hope, K., 1969. Methods Of Multivariate Analysis. Gordon and Breach,
New York: 288 pp.
2
Ito, K. and W.J. Schull, 1964. On the robustness of the T test in
0
multivariate analysis of variance when the variance-covariance
matrices are not equal. Biometrika 51= 71-82.
James, F.C., 197 1. Ordination of habitat relationships among breeding
birds. Wilson Bull. 83= 215-236.
Kessel, S.R. and R.H. Whittaker, 1976. Comparisons of three ordination
techniques. Vegetatio 32(1): 2 1-29.
Lachenbruch, P.A, 1975. Discriminant Analysis. Hafner press, Hew York:
128 pp.
Horris, J.M. and J.P. Barkham, 1 970. A comparison of some Cotswold
beechuoods using multiple-discriminant analysis. J. Ecology
58(3): 603-619.
Pimentel, R.A. and D. F-. Frey, 1978. Multivariate analysis of variance
and discriminant analysis. Chapter 9 in fiuantitative Ethology,
P.W. Colgan (ed.). John Wiley £ Sons, Inc., Hew York: 247-274.
Smith, R.W., 1976. Numerical analysis of ecological survey data. Ph.D.
dissertation. University of Southern California, Department of
Biological Sciences, Los Angeles: 402 pp.
Sokal, R.R. and F.J. Rholf, 1969. Biometry. W.H. Freeman and co., San
Francisco. 776 pp.
-------
APPENDIX A
DISCRIMINANT ANALYSIS (UNWEIGHTED) - SAMPLE CALCULATIONS
Data matrix

variable


1 2

1
.1 8 .
group 1
2
.3 5 .

3
.5 7 .
group 2
4
.7 6 .

Calculation of the pooled W matrix
1 . group means
variable
group 1
group 2
1	2
2	6.5
6 6.5
-------
584
APPENDIX A (CONTINUED)
2. group data centered (subtract mean value) by group maans
variable
1	2
1
. -1
1.5 -
C



1
2
1
-1.5 .

3
. -1
. 5 .
C



2
4
1
- . 5 .

3. W matrix for group 1
14 = C'C =
1 1 1
-1	1
1.5 -1.5
x
-1 1.5
1 -1.5
2 -3
-3 4.5
3. M matrix for group 2
W = C'C =
2 2 2
-1
1
. 5
- . 5
. -1	.5
x
1 - . 5
2
- 1
-1
. 5
5. pool
U = U + U =
1 2
2 -3
-3 4.5
2
. -1
- 1
. 5
-4
5
-------
585
APPENDIX A (CONTINUED)
C. Calculation of the T matrix
1 . over-all means
X = 4
1
X =6.5
2
2. data centered by over-all means
site
variable
1	2
1
-3
1.5 .
2
- 1
-1.5 .
3
1
. 5 .
4
3
-.5 .
matrix Z
3. T matrix
T = Z'Z =




-3
1.5 .

. -3
- 1
1 3

-1
-1.5 .
.20 -i+ .
.


X



. 1.5
-1.5
.5 -.5

1
. 5 .
. -4 5 .




3
-.5 .

-------
586
APPENDIX A (CONTINUED)
0. Calculation of the B matrix
1 . difference method
B = T - W =
16 0
0 0
2. direct method
e . g
b = SUM ( n ( x - x )(x - x ) )
12 h=1 h 1h 1 2h 2
= 2(2-4)(6.5-6.5) + 2(6-4)(6.5-6.5) = 0
-1
E. Eigenvalues and eigenvectors of 14 B
1. eigenvalue L =20
1
axis 1
2. eigenvector A
1
.78087
.62469
variable 1
variable 2
note s
Only one axis was defined since there are only 2 groups.
-------
587
APPENDIX a (CONTINUED)
F. Calculation of scores
S = X A =
1	1
1
8
3
5
5
7
7
6
x
.78087
.62469
axis 1
5.7784 0
5.46608
8 . 27722
9.21428
1
2
site
3
4
G. Coefficients of separate determination
1. put eigenvector in diagonal matrix of zeros off diagonal
78087	0
0	.62469

-------
588
APPENDIX A (CONTINUED)
2. calculate coefficients
D = Z TZ U =
111
78087 0
0 .62469
20 -4
-4 5
. .78087 0
0 .6 2469
10.243953
variable 1
variable 2
H. Standardised coefficients (by total SD)
1 . coefficient for variable 1 on axis 1
1/2
e = a (t /(n-1))
11 11 11
1/2
.78087(20/3) = 2.01620
2. coefficient for variable 2 on axis 1
1/2
e = a (t /(n-1))
21 21 22
1/2
.62469(5/3)	= .80647
-------
APPENDIX A (CONTINUED)
I. Standardized coefficients (by within SD)
2. coefficient for variable 1 on axis 1
1/2
f = a (u /(n-g ) )
n 11 n
1/2
.78087(4/2) = 1.10432
2 . coefficient for variable 2 on axis 1
1/2
f =a (w /(n-g))
21 21 22
1/2
.62469(5/2)	= .98772
-------
590
APPENDIX B
WEIGHTED DISCRIMINANT ANALYSIS - SAMPLE CALCULATIONS
A. Data matix - start with the same data as in the unweighted
calculations
variable
1 2

1
. 1
8
site
2
. 3
5

3
. 5
7

4
. 7
6
B. Matrix of weights tcould be from relative species abundances or
a GRSIM-type matrix, etc.)
site
12 3 4
group
1
. . 1
. 2
. 8
. 7 .
group
2
. . 8
. 9
. 1
. 3 .
-------
591
APPENDIX B (CONTINUED)
C. Expanded data matrix
Hote that there are four sites in each group (no site has a
weight of zero). Therefore the data matrix would appear
as such:
variable
2
2
3
4
1	8
3	5
5	7
7	6
group 1
site
2
3
4
1	8
3	5
5	7
7	6
group 2
-------
592
APPENDIX B (CONTINUED)
D. Calculation of the pooled W matrix
1 . weighted group means
variable
1	2
group 1 . 5.33333 6.44444
group 2 . 2.90476 6.33095
4
sun C y u )
i=1 i11 i1
ey' = 	
11
4
sun (u )
i= 1 i 1
H.I) + 3 t .2 ) + 5 ( . 8 ) + 7( .7 )
= 	 = 5.33333
. 1 + . 2 + . 8 + . 7
-------
593
APPENDIX B (CONTINUED)
2. center group data by weighted group means
variable
1	2
1
. -4.3333
1.5556 .

2
. -2.3333
-1.4444 .
group 1
3
-.3333
.5556 .

4
1.6667
-.4444 .

1
. -1.9048
1.6191 .

2
.0952
-1 . 3810 .
group 2
3
2.0952
.6191 .

4
4.0952
-.3810 .

3. W matrix for group 1 (W )
1
variable
variable
1	2
5.00000 -.66668
-.66668 1.044m
e.g. u = SUM C ( y - y* My - y' ) u )
12 1 i =1	ill 11 i21 21 i 1
(-4.3333X1. 5556H.1) + (-2.3333X-1.4 444X.2)
+ (-.3333 X .5556 ) ( .8 ) + ( 1 .6667 X-.4444X . 7 )
-.67409 + .67404 - .14815 - .51848
.-66668
k.
-------
594
APPENDIX B (CONTINUED)
4. M matrix for group 2 (W )
2
v ar iable
variable
1	2
8.38096 -2.92381
-2.92381 3.89524
4	_	_
e.g. w = SUM ( ( y - y' My - y' ) u )
122 i = 1	i12 12 i 2 2 22 i2
= C- 1 . 9048 ) ( 1 . 6 1 9 1 ) ( . 8 ) + ( . 0952 )(- 1 .38 1 M .9)
+ (2.0952M.6191M.1) + ( 4 . 0952 ) (- . 38 1 ) C . 3 )
= -2.46710 - .11832 + .12969 - .46808
= -2.92381
5. pool
W = W + U
1 2
variable
1	2
1 3 . 38096
-3.59049
•3.59049
4.93968
variable
-------
595
APPENDIX B (CONTINUED)
E. Calculation of the T matrix
1 . over-all weighted means
a. for variable 1
2 4
SUM SUM ( x u )
h=1 i=l ilh lh
x'
1	2 4
SUM SUM ( u )
h= 1 i= 1 ih
1 ( . 1 ) + 3 C .2 ) +5( .8)+7( . 7 ) + 1 ( . 8 ) + 3( .9)+5( . 1 3 + 7 ( .3)
.1+.2+.8+.7+.8+.9+.1+.3
15.7/3.9 = 4.02564
b. similarly, for variable 2
_ 8( . 1 ) + 5 ( . 2 ) + 7( . 8 ) + 6 ( . 7 ) + 8 ( . 8 ) + 5 ( .9 ) + 7 ( . 1 ) + 6( .3)
x. = 	
2	3.9
= 6 . 4 1026
k
-------
596
APPENDIX B (CONTINUED)
2. center data by over-all weighted means
variable
1	2
1
. -3.02564
1.58974 .


2
. -1.02564
-1.41026 .
group
1
3
.97436
.58974 .


4
2.97436
-.41026 .


1
. -3.02564
1.58974 .


2
. -1.02564
-1.41026 .
group
2
3
.97436
.58974 .


4
2.97436
-.41026 .


3. T matrix
variable
variable
1	2
19.09744 -3.44102
-3.44102 4.94359
2
e.g. t = SUM
12 h= 1
4
SUM < ( x - x'
i=1	i1h 1
) ( x - x» ) u )
i2h 2 ih
= (-3.02564) C 1 .58974)(. 1 ) + (- 1 .02564 ) (- 1 .4 1 026 ) ( .2 )
+ (.97436 )C .58974)(. 8 ) + C 2 . 97436 ) (-.4 1 026 )( .7 )
+ C-3.02564 )C 1 . 58974 ) ( .8) + < - 1 . 0 2 5 6 4 ) ( - 1 . 4 1 0 2 6 ) ( .
+ (.97436 )(.58974 )(. 1 ) + (2.97436 )(-.4 1 026 )( .3 )
-3.44 102
-------
597
APPENDIX B (CONTINUED)
F. Calculation of the B matrix
1 . difference method
variable
1	2
B = T - U =
5.71648 .14947
. 14 9 47 .0039 1
variable
2. direct method
2	_
e.g.	b = SUM ( p ( y' - x' ) ( y' - x' ) )
12 h=1 h 1h 1	2h 2
= ( 1 . 8M 5 . 3333 3-4. 0 2563)C 6 . 44444-6. 41026)
+ (2.1)(2.90476-4.02564)(6.38095-6.41026)
= .14944 (differs in the fifth decimal
from the difference method -
due to rounding error)
G. Once the U, B and T matrices are calculated, the analysis proceeds
as with unweighted discriminant analysis. When calculating the
scores, use the original unexpanded data matrix, since only
one score per site is required.
-------
Intentionally Blank Page
-------
5tii!r Ciili'on»tsj
Ni e m orandum
THE S;.5CU3CCS AC
To : Mr. L. Frank Goodson
Project Coordinator
Resources Agency
1416 Ninth Street
13th Floor
Sacramento, CA 95814
Prom : STATE WATER RESOURCES CONTROL BOARD
DIVISION or WATER QUALITY
Select; REVIEW OF DRAFT ENVIRONMENTAL IMPACT REPORTS (ElR's) STATE CUARIHGliOUSE
NO. 79C51509A, FOR CITY OF LOS ANGELES, TERMINAL ISLAND TREATMENT FLAKf,
PROJECT NO. 1202
«
This office has reviewed the two .draft E/R's for this project ar,d appended
report, dated May, 1979. The Division of Water Quality hereby presents
preliminary comments on th? draft documents.
0
State Clearinghouse No. 79051509A includes a Draft Environmental Irr,pact
Report (EIf<) on the proposed new effluent disposal system for Terminal
Island Treatment Plant, a wastewater treatment plant of the City of
Los Angeles. The recommended alternative as a remanent effluent disposal
system is an outfall to Los Angeles Harbor in Snn Pedro Bay. Its
acceptance as a permanent system relies upon acceptance by !:ne Regional
Water Quality Control Board of the concept that the effluent enhances
the Harbor waters. The alternative presented is an ocean outfall,
constructed essential ly as an extension of the harbor cu'cfall. S-^r.e
the present outfall site may soon be buried under a landfill, the City
has also presented a second EIR under the same Clearinghouse n^bsr.
It describes the Harbor outfall as4 if necessary, a t^mpcr^ry solution,
that can be extended to the ocean if the claim of enhancement is
rejected. This procedure was necessary to ..prevent possible delays of
a Clean Water Grant to construct the harbor portion of the outfall, on
the presumption, that the existing outfall might be burif'J before fii:al
determination of-the issue of enhancement. The format of the two
EIR's is similar.
The third document under review as Clearinghouse Nc*. 75G::i?j09A h Part. 10
c-f the Marine Studies of San Pedro Bay. It is incorporated by roforcncs
under both the above cIR's, and was issued along with them. Cur comments
on this third document will be submitted in a separate lettc-r.
Terminal Island.Treatment Plant Unit II C, Effluent Disposal Sy>to-:n Craft EIR
Sursnary Background
Comment: The Bays and Estuaries Policy that forbid:; discharge of affluent
to San Pedro Bay (unless enhancement is demonstrated) is a policy
of the State Water Resources Control Board, ;>ot EPA•, «r is
correctly stated on. Page 11-4.
c~'
Reproduced from
best ava'l^ble copy^
-------
Mr. L. Frank Goodson	-2-
Alternatives
II.B.3.f.(1)	- Federal Water Pollution Control Act - 197? Amendments
Comment: This section is out of date. It should describe the Clean Water
Act of 1977, P.L. 95-217.
III.C.3.	- Environmental Setting, Project Site, Effluent Characteristics
Comment: The EIR points out that numerous operational difficulties occurred
at Terminal Island Treatment Plant during 1978 and that "...in-
dustrial discharges were apparently the major cause of the large
number of violations in the second half of the year," because
"...certain industrial dischargers were violating the conditions
of their discharge permits,..".
Moreover, it is pointed out in Section IV.A.2. (Page*1V-1) that
industrial wastewater formerly discharged to the los Ancelcs
Coimty Sanitary Districts' system now enters Terminal Island
Treatment Plant. The EIR points out in Section IV.A.5.a. (see
below) that photosynthetic activity in the harbor was lower
in 1978, although there was no drop in phytopiankton. Did the
Terminal Island Treatment Plant influent during 1978 contain
newly added tone constituents {such as mi rex or other chlorinated
crganics used as fire retardants) that might have inhibited
photosynthesis?
111 .D.5.d. - Water Column Fauna
Comment: It is unclear when the settling rack studies described were performed,
We understand that such studies were funded for 1978. Please clarify
what is described. •
III.D.5.e. - Environmental Setting, Ecological Systems, Marine Biology,
Benthic Fauna
Comment: The section describes changes in species numbers and biouass
in general terms, and asserts a rapid recovery in these
parameters during the 1978 plant upset. Was the rapid
increase due to recruitment of those opportunistic species
(such as capitellid polychaetes) that are commonly
asserted to be indicators of a stressed environment? C?r,
arty qualitative judgments be made about spatial differences
in benthic fauna?
111.0.5.f. - Fish Fauna
Comment: Paragraph 1, Page III-44, states that "Both of these species
(anchovy and white croaker) fed on the particulates...and on...
benthic worms...". What evidence shows that anchovies eat
benthic worms?
ir .-r\
-------
\
Mr. I. Frank Goodson
III .D.!5.f. - Fish Fauna
Comment: (cont.)
This section seems to be based entirely on the results of the
Harbors Environmental Projects (HEP) surveys. How do the
results cf other studies, such as those performed for Southern
California Edison Company, compare with the HF.P surveys?
Comment: If the kelp bed planted on the Middle Breakvrater "did not develop
¦* /» A WO	k -1	n V>«\ n/\4-	1 A ^ V*	K	r U A I I 1 *4 f U 4 f «/* '- t.n	n *4
for reasons that are not clear...", should this not be cited
as possible evidence of toxicity of Terminal Island Treatment
Plant "effluent, considering the evidence that the effluent may
suppress photosynthesis?
III.D.5.3•	- Marine Mammals
w— 0
Consent: Does the statement	that sea lions, seals and dolphins "...are
not dependent upon	the Habor for their livelihood" mean that
loss of fish fauna	has no impact on marine manuals?-
IV.A.5.e.	- Long-Term Operational Impacts
Comment: Chlorination is stated to have occurred March 9 - Aucust 30, 1978.
rt • 		_ .n • _	' it. li._ _ x . j.	a.	r •	_ t \; n nrLi	_•	x. •	
This conflicts with the statement on Figure IV-4 "Chlorination
February - August." This is an important point in evaluating
impacts. Please give dates of chlorination.
The statement is made (Page IV-II) that "the research shoys that
there 1s a beneficial impact associated Kith the discharge of
biodegradable wastes-into "the harbor." The statement should
also call attention to the evidence that points out the possibility
that Terminal Island Treatment Plant effluent seriously inhibits
photosynthesis in the harbor. The account of. that critical
point 1s limited to the statement.later in the same section
(Page IV-13) that "...productivity and assimilation ratios...are
drastically reduced, presumably due to loss of nutrients, or
to inhibition." In our view, the evidence presented in
Reference IV-6 (the ecological study don» for this project)
makes nutrient limitation improbable. The EIR must edrnit the
evidence for adverse water quality impacts along with the
emphasis on bio-enhancement.
The statement (Page IV-14) that "...loss of bacterial populations
will also be reflected In the ability of the harbor to assimilate
wastes..." seems to ignore the rapid rate of reproduction of
bacteria. Is there any actual ev.idsnce to supprt the statement?
III.0.5.g. - Algal Flora
CT9
Reproduced from ||w
best available copy.
-------
Reproduced from
besl available copy.
fir. L. Frank Goodson	-4-
IV.A.5.e. - Long-Term Operational Impacts
Connent: (cont.)
On Page IV-15, the EIR states "Biostimulation and grov.th
experiments iri the field and laboratory shewed that both
pre-DAF cannery waste and Terminal Island Treatment Plant
secondary waste could sustain or stimulate growth in
phytoplankton, some invertebrates, and some fish." We
have difficulty finding experiments that demonstrate, for
instance, effect of^Terminal Island Treatment Plant
secondary wastes on"growth of fish. Please point out the
appropriate experiments in Reference IV-6. .
Conclusion (Pages. IV-21-22)
Comment: The EIR states (Page IV-21) ifiat "...release of managed levels *
cannery wastes into the harbor without secondary treatment of
those wastes would create a better nutrient balance in conjunct*',
with secondary Terminal Island TreatiTimt Plant wastes...". Is
it the City's position that cannery wastes should be removed
from the treatment plant? What is meant by "a tetter nutrient
balance?"
IV.B. - Mitigation Measures Proposed to Minimize Environmental Impact
IV.B.4. - Protection of Endangered Species
Comment: The EIR lists two possible mitigation measures to protect nesting
sites of Least Terns.- A mitigation measure must be chosen and
put Into effect as a condition of grant funding for the project*.
Terminal Island Treatment Plant, Unit II C, Harbor Outfall Draft EIR
Summary - Environmental Setting Page J.-3
Comment: The statement that "the Terminal Island Treatment Plant effluent
is the only remaining nutrient source to the harbor..." is
incorrect. As is pointed out elsev;bere in the EIR, large
amounts of ocean water enter the harbor daily. These waters
bear nutrients. Like all shallow marine bays, San Pedro Bay
maintains an intricate nutrient influx in its ecosystem,
including at least some recycling of nutrients from sediivent:..
Ill.C.3. - Environmental Setting, Project Site, Effluent Characteristics
Comment: Since the effects of chlorination may be critical to the environ-
mental impacts of the effluent, this cecticn should explain
whether or not chlorination of the	is J>; .
The discussion in IV.C.1. !.!•
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Mr. L. Frank Goodson
IjI.D, - Environmental Sstting, Ecological Systems
Cormient: Sections III.Q.e., III.D.5.f., and III.D.5.g. are similar to
the same sections in the Effluent Disposal System Draft EIR,
and our comments on those sections apply to both EIR's. The
problems relate to the claim of enhancement, which is asserted
not to be an issue in the Harbor Outfall EIR.
As a funding agency the SVIRCB reserves the right to make further comments
this project prior to granting EIR approvals pursuant to the Clean Water
Grant Regulations.
Should you have anv questions regarding this review, please contact
Howard Wright at (915) 322-7734.
nei i uiinnom ;
Division Chief
Manager - Clean Water Grant Program
cc: Mr. Russ Beckwith, EPA, Region IX
Mr. Lewis Shinazi, CRWQCB, Los Angeles Region ('*>)
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TMC RESOURCES AGE;
Mr. L. Frank Coodson
Project Coordinator
Resources Agency
1A16 Ninth Street
fcatei JUL 2 1973
In Reply Refer
to: 526:HOW
13th Floor
Sacramento, CA 95814
STATE WATER RESOURCES CONTROL BOARD
DIVISION OF WATER QUALITY
\ ~
REVIEW OF DRAFT ENVIRONMENTAL IMPACT REPORT (E1R), STATE CLEARINHOUSE
NO. 79051509A,- FOR MARINE STUDIES OF SAN PEDRO BAY, PART 16, ECOLOGICAL
CHANGES IN OUTER LOS ANGELES-LONG BEACH HARBORS FOLLOWING INITITATION OF
SECONDARY WASTE TREATMENT AND CESSATION OF FISH CANNERY WASTE EFFLUENT .
•
The subject document reports on part of a study, largely funded under a
Step 1 Clean Water Grant to the City of Los Angeles. The claim that, the
Terminal Island Treatment Plant (TITP) effluent enhances Los Angeles Harbor
rests primarily upon the subject document. The document includes a section .
upon the editor's views of the subject of enhancement. We do not choose to
comment upon that Section of the subject document. We have also not re-
peated comments that would essentially duplicate comments upon the two EIR's
that incorporated this document by reference (see our letter of 6/13/79).
Comment, Executive Summary
The statement is made on Page vii that productivity and assimilation ratios
"...are drastically reduced, presumably due to loss of nutrients, or to in-
hibition." Numerous statements throughout the volume, such as on Page-IIID7,
state that phytoplankton productivity is probably not nutrient-limited. If
that is true, the conclusion should be that there is evidence of inhibition.
Since the report is so massive, this critical issue deserves a separate sec-
tion, with graphical presentation of data on nutrients, rather than asking
the reader to draw conclusions from scattered statements or relying on the
Executive Summary.
The report states on Page ix that "The Iosb of bacterial populations will
also be reflected in the ability of the harbor to assimilate wastes, since
they were an important link in recycling material." Does this statement
imply that the present bacteria cannot reproduce quickly if more wastes are
added?
Comment, Section IB, Evaluation of Blocnhancc.mcnt in Outer Los Angeles Harbor
r
The criteria to be .used for enchancement do not include mention of levels of
primary productivity. Please state the reason for that exclusion.
Comment, Section HA. Changes In Fish Populations in Outer Los AnReles-Lons
Beach Harbors. Fish Trawla
This section seems to be based entirely on the results of the Harbors Envi-
ronmental Projects (HEP) surveys. How do the results of other studies, such
is those performed for Southern California Edison Company, compare with the
!KP surveys?	f ¦" 1	(Reproduced from
best available co
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Mr. L. Frank Goodson	-2-
Comment, Section IIA, Fish Populations, Conclusions on Fish Investigations
We agree 9trongly with the statements In this section that point out that
since the original drop in fish populations In the outfall areas predated
upgraded treatment, the dropoff therefore cannot be solely the result of
changes in effluent. This point should be reflected in the summary.
Comment, Section IIC, PhytoplnnUton Primary Productivity in Outer Los Angeles>
Harbor,"1976-1973
This section presents evidence that photosynthesis has either been inhibited
or limited by some change more or less coincident with the changes at TITP,
The normal seasonal increases in phytoplankton numbers were not, apparently,
accompanied by increased photosynthesis* The tables and figures are based
011 only six of fifteen stations, with no analysis of variance. Was the mean
assimilation ratio low in 1978 for* all stations sampled? Would an analysis
of variance show the 1978 drop in assimilation ratio to be statistically sig-
nificant for individual stations and/or for all stations sampled?
<
Comment. Section IIP, Changes in Zooplankton in Outer Los Angeles-Long Beach
Harbors, Conclusions
Tlie section says on Page I1D7 that, "...total concentrations are not greatly
increased...". Did concentrations of zooplankton increase or decrease? Were
any collections made that might answer this question, in view of the statement
(Page 1ID2) that changes in methods'exclude year-to-year comparisons of total
zooplankton?
Comment, Section HE, Changes in Benthic Fauna in Outer Los Angeles-Long Beach
Harbors, 1972-1978
This section concisely presents extensive data on changes in benthic popula-
tions. The only statement about qualitative differences is the statement on
page 11E9 that, "....species such as Capi tel la capitnta ... are hardy, oppor-
tunistic, fast-growing species that thrive in the absence of competition", and
that, "...the species occurs in many unstable (variable)- environments where
rapid growth and short, year-round reproductive cycles give them an advantage.
In view of the recent decline of such species, con the benthic environment be
inferred to be more stable now?
Is the sampling gear equally efficient in collecting small polychaetes and dec
burrowing clams and crustaceans that are less hardy and slower-growing?
Does the data on Station A1 (outside the harbor) indicate that the influence
of the harbor discharges on the adjacent ocean has decreased?
Comment, Section I1F, Fish F.gs and Larvae Surveys
This section presents data on temperature, salinity, and nutrients. Similar,-
more detailed figures would be very useful in a place easier for the reader
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Mr. L. Frank Goodson
to find. Scales that show the nutrients in milligrams per liter as well as
microgram atoms per liter, would be useful to decision-makers who must use
these results.	'
The statement (Page /IIF13) that "secondary treatment has not significantly
altered nutrient conditions" seems to conflict with statements elsewhere in
the volume. Please explain.	? ;	'
What "previous studies .show that the dominant adult fish Genvonemus utilized
the cannery area for foraging on suspended cannery wastes..."?
"The present study found higher counts of eggs and larvae than were "found in
an earlier study; this is probably due to reduced predation by adult fish
and to more efficient sampling methods." Why were methods so changed as to
prevent measuring changes in year-to-year abundance?
The section refers to greatly reduced numbers of anchovy eggs and larvae, but
omits direct presentation of the data. What quantitative statements can be
made about thfe decrease in abundance of anchovy eggs and larvae?
Contnent, Section 1VB, Weighted Discriminant Analysis of Benthic Data
This section contains valuable analyses of phytoplankton and other water
column measurements. A change in the title of the section would alert the
reader to seek such analyses here.
Comment, Section VA, Phytoplankton Growth and Stimulation in the TITP.Secondary
Waste Plume
The section on methods points out that results were highly variable with all
phy toplankton species used except Dunaliella tertlolecta. EPA work on D_;_ ter-
tlolecta (Proceedings of Seminar on Methodology for Monitoring the Environment,
Seattle, Washington October 1973) points out that that alga is well suited for
studies on nitrogen of phosphorus limitation. However, since it is tolerant of
toxicity and requires no outside source of vitamins, it is not suitable for
general assessment of unknown algal growth factors. Did results with other *
species point to apparent limitation by toxicity or vitamins? Could TITP ef-
fluent supply vitamins or metals that stimulate phytoplankton?
Did preparation and treatment of samples of TITP effluent allow chlorine to
escape before the growth test?	•
Since the laboratory scawater supply and artificial seawater apparently both
provided nutrient enrichment, why was open ocean water not used for dilution?
The last paragraph on Page VA7 seems to state that the effluent is diluted to
a IZ level 525 meters from the outfall, and a 27, level farther away. Is this .
an error?
€
Reproduced from
best available copy.
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Mr. L. Frank Goodson
-4-
Comment, Section VB1, TITP Secondary Waste Bionssays,, Discussion and Conclusions
The results seem to show toxicity that varies from one date to the next, but .
that is discounted in the discussion. Considering the fact that the effluent
varied greatly, and,was chlorinated on some dates and not others, is it not
reasonable to conclude that toxicity varied?
Examination of Tables 1, Z, 3 and U seem to show evidence that TITP effluent is
toxic to anchovy embryos. Are these data statistically significant?	>
Comment, Section VC, Cannery Waste as a Food for Anchovies
If the waste given the anchovies was sludge from the DAF units, is the study
.relevant to the situation prevailing during DAF treatment of wastes and dis-
charge to the hajrbor?
Were rigid glass rectangular tanks used, or were tanks chosen to allow circular
swimming patterns with miftiraal chance of contacting a hard suxface?':
*	0
As a funding,agency the SWRCB reserves the right to make further comments on
this report prior* to granting an EIR approval pursuant to the Clean Water Grant
Regulations,
Should you have any questions regarding this review, please contact Howard 0.
Wright at (916) 322-7734.

eil' Dunham
Division Chief
Manager - Clean Water Grant Program
cc: Mr. Russ Beckwith
EPA, Region IX *
San Francisco
Mr. Lewis Schinazi
CRWQCB, Los Angeles .Region (A)
Los Angeles
bees U^ike Fnlkenstcin
Ccrald Bowes, Planning & Rcaaarch
Mike Coony
William Attwatcr, Legal Division
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OF CAHfOVMlA—AGfNCY
^ARTMcNl Or FISH AND GAME
:ime resources region*
• Golden Shore
ig Beach, California 90802
.3) 590-5117 or 5118
EDMUND C . BROWN JR.. Go.cr
22 May 1979
RESPONSES BY HARBORS ENVIROMENTAL PROJECTS,
UNIVERSITY OF SOUTHERN CALIFORNIA, AS CONSULTANT
FOR THE CITY OF LOS ANGELES, TO THE LETTER BELOW
(RESPONSES INTERSPERSED IN ITALICS>-JULY 5, 1979
Mr. Jeffery D. Benit, Chief
Food Industry Branch (VH-552)
United States Environmental
Protection Agency
Washington, D. C. 20460
Dear Mr. Denit:
Review of report titled Ecological Changes in Outer
Los Angeles-Long Beach Harbors Since the Initiation
of Secondary Waste Treatment and Cessation of Fish
Cannery Waste Effluent at Terminal Island, California
submitted to EPA by Dr. Dorothy Soule. of the
Harbors Environmental Projects (HEP)	\
This is in response tc your request for review of the subject document.
We believe the philosophical, legal and biological issues pertinent to
this report merit significant consideration; however, in keeping with
the informal nature of the April 5 workshop, indicated in the cover
'letter you sent with the HEP report, we also offer these remarks as in-
formal or preliminary. Any Departmental position, if requested, would
be presented in a letter from our Director in Sacramento. With this
understanding, we offer the attached comments.
We believe the critique we offer addresses the more important issues
contained in the HEP document, but we do not consider it exhaustive.
We think there are other issues that merit discussion and reflected our
views on those lesser issues at our April 5 meeting. Since our meeting,
HEP.has published a final draft of the document. Because our comments
were delayed, we have elected to revise them somewhat to reflect changes
and additions to that document.
•
In closing, let me say that the HEP report, even with the imperfections
we perceive, represents a continuing worthwhile effort. In view of the ^
administrative and funding difficulties that have apparently accompanied
the accomplishment of this report, we think HEP's persistence is cer-
tainly commendable. And, although we do not always concur with conclu-
sions presented by HEP, we look forward to a continuing dialogue with
Dr. Soule and her colleagues.
Reproduced from
best available copy^
r/7
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Mr. Joffcry D. Denit, Chief
2
22 Mav 1979
We appreciate the opportunity to provide comment.
Sincerely,
cc: Dr. D. Soule, HEP
SWRCB - Div. Water Quality - Howard Wright
NMFS, Terminal Island (Jim Slawson)
USFWS (Jack Fancher)
RWQCB U (Dr. Lewis Schinazi)
EPA-SFO (Terry Brubaker)
Regional Manager
Attachment
C'"3
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California Department of Fish and Game Preliminary Review of Report
by the Harbors Environmental Project, Allan Hancock Foundation
(Dr. D. Soule, editor) regarding the Effects of Waste Discharges
on Los Angeles-Long Beach Harbors
Comment: The editors, D.F. Soule and M. Oguri, feel that so many items are
addressed which require correction or reply that the responses are inserted
in italics in a copy of the DFG review. June 15, 1979
Section IA - BI0ENHAXCEME3T: CAN THIS CONCEPT BE DEFINED AKD MEASURED?
The philosophical, and legal aspects of this question, used as the title
for the introductory section of the Harbors Environmental Project (HEP) report
critically important to the California Department of Fish and Game (DF&G)
because they may influence decisions made by Environmental Protection Agency
(EPA) California Regional Water Quality Control Boards (CRVOCBs)
4	J
and the California State Water Resources Control Board (SWRCB) re-
garding waste discharges which, in turn, may profoundly affect the accomp-
lishment of Dr&G's legislated mandate. Department of Fish and Game has
the unique responsibility to manage the state's fish and wildlife popula-
tions in addition to its charge to protect, maintain and enhance living
resources, e charge it shares with other agencies.
The principal question raised regards enhancement and pertains to the
Water Quality Control Policy for the Enclosed Bays and Estuaries of
California (The Bays and Estuaries Policy) adopted by the SVRCB. The
report tends to focus upon the lack of definition of the word "enhancement".
Although we believe this focus is important, there are several other points
that have been omitted from the discussion of the Bays and Estuaries Policy
and of enhancement. In addition, we believe the interests and functions
of the several agencies concerned with the Policy and the enhancement
Issues comprise an important perspective that oust be clearly under-
€ .0
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-2-
vhile these Issues are debated. Therefore we offer the following
review:
1. Misunderstandings by HEP personnel have occurred in the past at
public hearings and are apparent in the present document in dis-
cussion of the Bays and Estuaries Policy and its relation to law
and regulatory agencies. For this reason, we believe it is im-
portant for HEP staff and all others who may read these comments
to recall the path by which the Bays and Estuaries Folicy was
adopted and why EPA should be concerned with that Policy in its
thinking about discharges of seafood processing wastes to the
marine environment as the Agency carries out its duties required
by Section 74 of the Clean Water Act of 1977.
Contrary to statements contained in the introductory para-
graphs of the Bioenhancement section of the HEP's report, the
State Water Resources Control Boards and the Regional Water Quality
Control Boards were created in their earliest form when the
Legislature passed the Dickey Act in 1949-
Comment: The clear intent of the introductory statements made in Section IA
of our report is to outline the developments that led to the need to establish
a definition of "enhancement" as it appears in the document cited in our
report. It is impossible to establish the existence of such a condition as
enhancement unless there are criteria to establish what is to be demonstrated.
It was not our intent to document all of the legal milestones involved in
environmental law; clearly, the 1969-70 period was when the power of compliance
was delegated.
a. In 1969, the Porter-Cologne Water Quality Control Act
redefined the role of the SWRCB and broadened its powers.
In 1972, the Porter-Cologne Act was amended to enable
California to be designated by EPA to administer PL-92-500
€ .0
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-3-
in California.
b. Section 301C.3 of PL-92-500 sets forth the mechanism by
which state-enacted standards, when approved by EPA,
become the applicable federal standard for state waters.
The Bays and Estuaries Policy, adopted by the State
Board and approved by EPA, is an example of this mech-
anism. For this reason, and because the Policy specifi-
cally applies to the Los Angeles-Long Beach Harbor complex
not only the SWRCB but EPA must weigh the enhancement
issue when considering discharges to harbor waters.
Comment: The first paragraph of our report (without going into legal section
numbers) is directed at getting to the section of Bays and Estuaries Policy
under which the Regional and State Water Quality Control Boards were discussing
the cannery and Terminal Island sewage effluents. We did not intend to docu-
> ment the history of all environmental efforts at enforcement in the harbors
(e.g., since 1916, when complaints about sulfide fumes were recorded), but to
indicate the rationale on which the 1978 studies were based.
c. In 1974 the State Board adopted the Bays and Estuaries
Policy. It is important to note the Policy is not a
"plan" as defined by the Porter-Cologne Act. According
to Section 13050(j) a "Water quality control plan" must
designate 1) beneficial uses to be protected, 2) water
qxiality objectives and 3) a program of implementation.
This apparently is not the case for a "policy". Many
of the additions advocated by the HEP document
would convert the Bays and Estuaries Policy into a
"Plan" as defined above. In view of subsequent diffi-
culties with issues such as the definition of enhance-
ment, such a conversion may have merit.
^	C. "
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Comment: There is no place in our document, Marine Studies of San Pedro Bay,
California, Part 16, where we intimated anything about a "plan" instead of
a "policy". Nor have we been confused over the difference. Rather, we
described some of the "evolution" (p. IA5) exhibited in the stated goals of
various plans for the harbor, solely in order to make the point that no
resource-oriented environmental goals were even expressed for organisms or
habitats until very recently; only "people" goals were spelled out.
2. The stated purpose for the Bays and Estuaries Policy, presented in
the first introductory statement of the Policy is "...to provide
vater quality principles and guidelines to prevent water quality
degradation and to protect the beneficial uses of enclosed bays
and estuaries" (emphasis added). We believe this statement
clearly indicates that vhen it adopted the policy the SWRCB did
not intend to create a plan but acted only to declare its policy
intentions and left the prescribing of mechanisms for achieving the
policy for Regional Boards to accomplish.
Comment; Authority was delegated by EPA to the State of California Water
Quality Control Board and the Regional Boards to make determinations and issue
permits for effluents. Presumably this meant a better measure of local control
and on-site information. The Regional WQCB, as a matter of record, found in
1976 that enhancement was occurring; but this finding was twice rejected by
EPA Region IX, as we recall. The RWQCB was ordered to withdraw their drafted
permits for the cannery effluents, and the Terminal Island Treatment Plant
was permitted a year of secondary effluent emission to monitor and attempt
to determine whether enhancement existed under the new conditions. There was
no "Plan" involved, but clearly a "Policy" was involved, which is stated;
.however, it has not been possible as yet to define the word "enhancement"
biologically, to EPA's satisfaction. A draft of the chapter on bioenhancement
was circulated to a number of scientists and the RWQCB for comment several
months before the April draft was assembled.
It is clear that the law, as it exists at any given moment, is subject to
enforcement. However, laws are usually general, and regulations are created
to implement the laws with specifics. Regulations can and should be changed
in the light of new information; so can the law, although it is admittedly
a much slower process.
But, there must be provision for feedback in the Regulatory hierarchy. Costle,
jorling and Davis of the EPA have all made speeches about the desire of the
Agency for input from the public (I assume the scientific community is at
least included in that category). Jorling has recently asked the rhetorical
question as to whether the Agency is capable of flexibility, and of mid-course
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-5-
correction in the light of new evidence,— or is the Agency too monolithic
to adjust? If there is no connection between the administrators and the
local regulatory groups, then no changes in regulatory posture occur in
spite of speeches made.
' 3. The State Board declared its intention to be that "...the dis-
2/
charge of wastewaters and industrial process waters...— to en-
closed bays and estuaries....shall be phased out at the earliest
practicable date." (Chapter I Section A). We believe two points
need to be made about this statement.
a. The issue of enhancement applies to the exception rather
than the rule. This indicates to us that the demonstra-
tion of enhancement oust be unequivocal.
Comment: The comment concerning the definition of enhancement again applies
here. How can enhancement be declared to be unequivocal if nobody will issue
a definition and criteria for measuring the conditions?
Any good biologist would surely qualify his/her remarks, as there are no abso-
lutes (unequivocals} or guarantees in biology. It is patently impossible to
come up with any biological statement that can not be argued to some extent
by peers.
There are no other regular organized studies of the same harbor area in ques-
tion, covering the same lengthy time period. Reference to other areas of the
harbor (Edison) or other sites in California, or random diver observation can
not be applied to the site of the precise area (case study) to which the
question of bioenhancement was directed.
b. Footnote 2 refers to the discretion that RWQCBs have
regarding implementation of the Policy. We believe this
~
again indicates the State Board's intention to provide
general guidance only and leave specific determinations
regarding enhancement for the Regional Boards to decide.
Comment: if the footnote, as referenced, delegated the decision-making to the
RWQCB, why have they been forced to withdraw the drafted permits because EPA
Region IX objected to them?
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-6-
4. We believe the SWKCB clearly indicated why discharges should be
elininated from bays and estuaries ar.d why the demonstration of
enhancement should be clearly beyond question before exceptions
are granted. For example, Appendix A to the Bays and Estuaries
Policy is an analysis of comments received by the State Board
prior to its adoption of the Policy. On page 5 of Appendix A,
in the Discussion section, is the following statement:
"The enclosed bays and estuaries of California have
values to the people of the State far in excess of what might
be expected on the basis of their size alone. Aside from the
San Francisco Bay Delta system, they are fairly small isolated
features which presently receive modest quantity of municipal
and industrial wastes (See Exhibits C and D). In view of the
value of this resource, the low wastewater flows which are pre-
sently discharged, and their proximity to open coastal waters,
the staff believes that it is both desirable and feasible to es-
tablish a policy in which the criteria for discharge is water
quality enhancement rather than maintenance.
"Experience has shown that within a wide range of treatment
alternatives, ocean discharge is generally preferable to estu-
arine or bay disposal. We have a particularly good example of
this in California (i.e., San Diego Bay)."
•
Comment: The last sentence of the second paragraph uses the words "... water
quality enhancement rather than maintenance." The root of the problem has
continued to be that a dictionary definition, such as suggested in the last
paragraph of your review (p. 5) does not deal with biological resources; it
deals with "people goals".
Consider another Webster dictionary definition of enhancement: "to advance;
to augment; to increase; to heighten; to make more costly or attractive; ...
to render more heinous; to aggravate ..." Wot particularly relevant to the
€
A
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problems at hand, but open to a wide variety of applications. Criteria are
essential.
With the above review in mind, we wish to return to the HEP's discussion
of the enhancement issue. Considering a similar case, regarding the en-
hancement issue and municipal discharges from the City of Areata to Humboldt
Bay, the HEP document states that a letter on the subject, written by
Mr. Bill B. Dendy, Executive Officer of the SVRCB, goes beyond footnote 3
of the Bays and Estuaries Policy because he stipulated uninterrupted pro-
tection. We disagree. Footnote 3 provides for percent-survival tests as
a measure of enhancement, as the HEP document notes; however, the footnote makes
three additional and, in our opinion, important statements:
1)	Maintenance of survival alone shall not constitute a demonstra-
tion of enhancement.
2)	"Full and uninterrupted protection for the beneficial uses of the
receiving water must be maintained.15
3)	Regional Boards may require physical, chemical, bioassay and
bacteriological testing prior to authorizing discharge to en-
closed bays or estuaries.
We believe these statements place the full responsibility upon
Regional Boards for deciding enhancement issues and we think there is
wisdom in allowing Regional Boards to make primary decisions governing the
0
waters within their purview, with a higher level of review vested in the
SWRCB to resolve conflicts that may arise.
Comment: Note that the Boards are supposed to make decisions.
f: ~
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In view of the "full and uninterupted protection" provision stated
In Footnote 3 and reaffirmed in Mr. Bendy*s letter, we cannot concur with
the statement on page 5 of the HEP document which says "....it seems
desirable that, in semi-enclosed bays and harbors, some averaging con-
ditions should be allowed over space."
Comment: This is HEP's stated opinion.
Ve believe enhancement can and should be defined. As a point of
departure, we believe a dictionary definition is appropriate: (i.e. to
increase or make greater as in value, beauty or reputation; augment. To
improve or make better). However, we believe the determination of en-
hancement should be on a case-by-case basis. .We suggest that one way to
accomplish this task would be for the Regional Board to appoint a committee
consisting of representatives from regulatory agencies, academic institu-
tions, and industry (i.e., the discharger). The committee would have at
least the following tasks:
1)	Meet before the enhancement demonstration is undertaken to deter-
mine the criteria to be satisfied and to evaluate study plans
proposed by the affected discharger.
2)	Monitor the progress of studies and suggest appropriate modifi-
cations.
3)	Evaluate the studies upon their completion and make findings and
~
recommendations to the Regional Board.
Comment: With regard to establishing a committee to design the investigations
prior to the study: The fact is that HEP put public information above pro-
prietary contracts and was able to obtain a baseline that no_ single agency
funded, over a long period (1970-1979).
The scope of work for 20-30 contracts and the federal Sea Grant Program were
designed with the needs of the various funding agencies or entities in mind
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9-
and to meet federal, state or local requirements. The procedures used are
acceptable worldwide. The early VSC Sea Grant Program (1971-75) was reviewed
by national panels and selected specialists, and was subjected to annual site
visits of 20-30 people as well.
The 1976-78 studies were planned with the active participation of the Regional
and State WQCB; the field work was carried out with Regional and State WQCB
people on board the research vessels, as well as representatives from the
Ports of Los Angeles and Long Beach and the City of Los Angeles Public Works
environmental staff.
Review copies of our report were sent to academic institutions across the
country. The Washington office of the National Marine Fisheries Service has
circulated many copies of the summary and Mr. Leitzell, the Director of NMFS,
has made the study of effluent enforcement a priority item for an advisory
subcommittee as well as for his Habitat Protection division. The Science
Advisory Board of EPA has also initiated examination of effluent guidelines
as a priority.
The DFG resources and environmental staffs were supplied with copies of the
full volume as soon as it was delivered by the printer. A partial draft was
courier-serviced to Mr. Dysinger on February 7, in an attempt to provide him
with research results which EPA had said they were unable to fund as part of
their Report to Congress on seafood processing effluents. It was probably
gullible of us to expect that any EPA personnel might view the document as
a resources study and not something which they considered "illegal" before
the document was even written. EPA apparently regards any flexibility on
effluent regulation as leading to the "domino effect" on EPA control, without
regard to whether mid-course correction is needed to save the environment
rather than to save the bureaucracy.
We agree with the suggestion in the HEP document that several criteria
should be applied to a determination of enhancement, but we suggest that
the concept of protection of beneficial uses (as defined by lav and basin
plans) be included in those critera. Generally, we support the idea of an
enhancement demonstration for gaining exemptions from the Bays and Estu-
aries Policy provided:
1.	A beneficial use is created or enhanced.
2.	Waste discharge requirements can be set to assure full and unin-
terupted protection of existing beneficial uses.
'3. Continued discharge to bays or estuaries would not cause con-
flicts in the uses that could be made of the receiving waters in

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-lO-
th e absence of the discharge.
4. Benefits accrued from allowing a discharge to bays or estuaries
do cot compromise fish and wildlife resources or preserves.
Comment: The HEP harbor research clearly showed a decline in benthic organisms,
fish and birds that cannot be explained away by DFG statements that the animals
are Just more normally random in distribution. Even cursory examination of
the 1977 data reported in the Southern California Edison study (EQA-MBC 1978)
shows incredible oscillations in the fish populations near the intermittently
flowing warm water discharge. Figures are attached to demonstrate this.
A report of 3385 dead California gulls in outer harbor waters due to an oil
spill would rightly be viewed with alarm and horror by DFG as an environmental
insult. But the HEP report that there were 3385 fewer birds per observation
day, as documented by an eminent ornithologist, was considered by DFG to be
equivocal or of no significance. DFG cannot decry the loss of California Gull
nesting sites at Mono Lake (Los Angeles Times) even at a cost of depriving
thousands of people of drinking water, after having said that a 23-fold decrease
of California Gulls in the harbor is just a loss of garbage-eaters (Long Beach
Independent) and that gulls eat Least Tern eggs'. We have found no reference
to support the contention that gulls do damage the Tern population by eating
Least Tern eggs in the harbor. It was documented in the HEP report and the
newspapers that humans (with trucks or bulldozers) disturbed Least Tern nesting
on Terminal Island in previous years and that Least Terns did achieve nesting
success when undisturbed by people.
Summary and Conclusions
The remarks offered below are intended not only to recapitulate points
ve believe to be important but to respond to questions contained in the
"General Review Criteria" section of the L.A. "Harbor Enhancement Study
Review Criteria enclosed as EPA guidance for reviewing the HEP docu-
ment .
1. Generally, ve believe there is" a place for both managed artificial
environments and natural and pristine environments. Managed
environments, in our view, imply hatchery operations for the
case at hand where nutrient inputs, water quality, and wastewater
quality are carefully and completely controlled. Hatcheries and
C .3
j
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other aquaculture operations have a valid place in the husbandry
of living marine resources, both as food sources for man and for
the enhancement of a species for its own sake. We do not, however,
concur with the concept implied in this document and advocated
elsewhere by HEP staff for an "open aquaculture" system especially
in bays and estuaries because they offer the potential for un-
«
desirable eutrophic conditions and because they do not provide
for strict controls of such factors as water quality or nutrient
inputs of proper type in proper amounts, nor do they assure that
desirable (i.e. for market or sport) fish species will be harvested.
On the other hand, natural nursery grounds and refuges that
estuaries or bays provide should be preserved. Ve believe the
Bays and Estuaries Policy in its present form will help attain
this objective.
Comment: Hatchery environments have been the particular expertise of DFG for
many years, because the original mandate in part was to provide freshwater
anglers with fish to catch. Hatcheries for marine species are in their
infancy by comparison, and closed-system, single-species aquaculture has
encountered many difficulties with mass infections and mortalities, and poor
flavor or texture in some final products. The so-called open aquaculture
system is a permitted function under EPA regulations (318) for semi-enclosed
basins, but so far it seems to have been used only to enable power plants to
enlarge their bundaries for thermal compliance and perhaps to raise gourmet
lobsters.
Comment: With regard to the lack of ability to manage effluent nutrient loads,
HEP has worked on computer models for several years to move toward managing
a semi-enclosed body of water. We were, in fact, successful in working with
the canners between 1975-1977, to maintain water quality. StarKist Foods limited
their reduction plant processing on the basis of monitoring, sometimes daily
and sometimes twice a week, for BOD loading and BOD and DO in the effluent
plume. Temperature and seasonal meteorologic and oceanographic conditions must
also be taken into consideration, based at this point on expertise, not on
modeling. There were no more episodes of zero DO in the harbor after we began
to work together. There also have not been any more harbor-wide red tide
episodes since 1973-74, although no one knows what the triggering mechanisms
are for dinoflagellate blooms. Phytoplankton still remains higher than normal
in Long Beach almost all year long, particularly in the area of the receiving
waters for the Los Angeles River, around Pier J, and in 1978 in the Long Beach
Middle Harbor and Back Channel.
CD
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CommentWhile people consider clear water to be esthetically pleasing, NMFS
scientists and others have shown that larval Jfish need turbid waters to cut
down on predation and to provide sufficiently dense phytoplankton for contin-
uous feeding. Therefore, a compromise in favor of the larval and juvenile
fish in the harbor would seem to be appropriate, since clear waters for swim-
ming and water-skiing among the tankers and freighters are not considered
appropriate uses of most of the harbor. There are, of course, and for years
have been, swimming areas at Cabrillo and Long Beach in the harbors.
Comment: The harbor has been eutrophic only in the sense that it is not nutrient-
limited for organisms. It has not been eutrophic in recent years in the sense
of supporting algal scums such as happens in freshwater lakes and streams.
Beginning in 2969-1970, when legislation put the power of enforcement into
pollution control (even if the DFG reviewer believes that it was possible earlier),
the harbor improved dramatically. This was documented (Reish, 1971). Raw
sewage under every boat and dock, fish cannery scrap and industrial wastes from
refineries certainly provided excess nutrients, but depleted the oxygen severely.
Once oxygen returned to anoxic areas of the harbor, blooms followed, but so did
a wide variety of organisms.
Comment: If HEP advocated dumping toxic chemicals into the harbor, or making
irreversible alterations to the marine waters, it would rightly be viewed with
alarm. That is not the case. All we have suggested and recommended are large-
scale experiments, in the outer harbor instead of in the laboratory, of con-
trolling effluent quality and quantity. Surely it is not such a drastic,
dangerous, threatening proposal to try something a little different to see
whether we can build up inshore fish populations once again. If computer tech-
niques, monitoring and good management of effluent levels don't produce the
desired results, or if degradation results, the permits can easily be revoked.
An adequate test period would perhaps resolve the entire question.
In many places around the world inshore fish populations have declined; where
salmon carcasses no longer decay in Oregon rivers and cannery effluents have
been shut off, larval salmon apparently don* t survive in adequate numbers; it's
in the literature. We repeat, What is so perilous about trying an open
aquaculture experiment?
2. Enhancement may well be a valid reason for granting exception
to a policy prohibiting waste discharge to enclosed bays and
estuaries, but the criteria for evaluating enhancement as
defined in the HEP document are inadequate for several reasons,
including but not limited to the following:
a. The criteria do not include protection of beneficial uses
defined by law and in current Basin Plans. Instead, the
cr:o
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beneficial uses defined by the Los Angeles RWQCB in 1969
are listed (on page 5) and criticized because they were
"based only on human orientations..." and "...no mention
of natural or biological environment was made."
In contrast, we note that beneficial uses specified
in tlit* Rl/nCG's rurront flQ7fi1	di , r t
viy/o; basin PJan for the Los An^i'Ig
Long Beach Harbor complex induce:
Industrial water supply
Navigation, commercial, and naval shipping
Water contact^recreation (e.g. swimming)
Non-contfo-L water recreation (e.g. boating and fishing)
Commercial fishing
Preservation of rare or endangered species
Preservation of marine ecosystems including propagation
and sustenance of fish, shellfish, marine mammals,
waterfowl and vegetation such as kelp.
Because the beneficial uses do include consideration for
biological values, we believe they are a valid criterion for
determining enhancement.
Comment; There was absolutely no suggestion in the HEP document that the other
uses of the harbor were not valid. Page IA5 of our report presents a dis-
cussion of beneficial uses and suggests the possible expansion of these. It
has been the HEP observation that agencies do not seem to recognize that one
of these beneficial uses is the use of the harbor as a delivery system for
the 10 million people in Megalopolis. The Coastal Commission, DFG, and F&W
have, on the contrary, generally applied intertidal open coastal habitat
criteria to harbor pilings, to the dismay of port authorities.
»
b. Although the criteria do distinguish between benefits to nan
and perceive benefits to the biota for their own sake, they
do not iaake distinctions between evaluations based upon
biological parameters (e.g. species diversity) and those
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-14-
based upon political, legal or other parameters based upon
human value judgement (e.g. species with sport or commercial
value; designation and definition of rare and endangered
species). We believe it is important to distinguish between
enhancement that assures full and uninterrupted protection of
nursery grounds, for instance, and enhancement that makes
desirable sport or commercial fish species more convenient.
Comment: The scope of work of the TITP EIR or any other biological investiga-
tion did not at any time call for HEP to do research on political, legal or
human value criteria. We discussed them as the introductory frame of reference
to establish why the field and laboratory research were needed and what uses
would be made of the document.
c. The criteria for enhancement favor an "averaging" concept
rather than the full and uninterupted protection clearly
proclaimed in the Bays and Estuaries Policy.
3.	We believe that marine ecosystem management programs incorporating
inputs of wastewater as a major component of the plan should be
confined to closed, carefully controlled systems such as hatcheries
or, at the most liberal extreme, in marshes created or augmented
by carefully controlled wastewater flows.
Comment: Waste water in a marsh may in some cases do more damage than good because
the mineralized nutrients in secondary waste water force the system toward
eutrophication.
4.	We believe the work presented has omitted meaningful comparisons
with other similar enclosed bays such as San Diego Day. Waste
discharges for example have been largely eliminated from San Diego
Bay. Preliminary data gathered by DF&C personnel monitoring fish-
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15-
eries populations in that Bay indicate that
the spiny lobster (PanuljrHs interruptus), a species historically
recorded throughout the Bay is slowly returning after a period of
absence presumably linked with waste discharges to the Bay.
Comment: HEP does not feel that San Diego Bay has had an adequate, long term
baseline study suitable for comparison with the HEP baseline and research
carried out for the City of Los Angeles TITP SIR study. The reviewer ignores
the mandate of the TITP study. San Diego Bay is a very different hydrological
entity, with severe flushing problems due to piecemeal dredge and fill and
multiple runoff sources. Fish cannery wastes were never examined as a part of
the San Diego Bay system. We suggest that a study similar to ours be carried
out by DFG to ascertain whether or not the San Diego Bay is similar to Los
Angeles-Long Beach Harbors.
The spiny lobster may 2>e returning because of removal of San Diego Bay sewage,
and/or because the role of surf grass as the habitat for the smallest larvae
has been identified, and/or because of oceanographic conditions, flushing by
rainstorms, the temperature and duration of the winter countercurrent, and/or
because DFG has enforced size limits on SCUBA divers' take. It has little
bearing on the lobsters found on the Federal Breakwater in Los Angeles-
Long Beach , or on the conduct and results of the legitimately scoped research
by HEP on the TITP study. Divers found a 15 lb. lobster on the Federal Break-
water in 1977; no conclusions can be drawn from that, either. We have avoided
announcing their presence, to avoid depredation.
Did DFG in fact measure water quality and BOD, TOC, TVS, etc. before and after,
and carry out a sampling program suitable for making the assumption that the
lobsters in San Diego Bay were linked to wastes? Was their scope of work
designed by a committee and supervised by an on-site advisory group?
As referred to in the critique, there is no indication that the "population
dynamics" criteria, which were not otherwise identified, used to criticize
our report are available or were thought of in considering the San Diego
lobster population.
5. We believe that cautions offered by Calaprice (1979) ~ are valid.
He states that while biostitnulation has occurred, only a small
fraction of the energy of organic matter discharged to the marine
environment in sewage sludge has been used through the harvest
of marine species.
New Comment: Only a small portion of an% energy level is transferred to other levels.
1/ Calaprice, J. R. 1979. Draft Summary Report. Evaluation of Marine
Impacts of Sludge Discharge to the Ocean under Baseline Conditions.
Report to the Los Angeles/Orange County Metropolitan Area Project for a
Regional Wastewater Solids Management Program (LA/OMA Project). Pfi- 41
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BIOLOGICAL RESOURCES
yiiTTnnArv of Comments
The remarks chat follow are offered in response to the EPA
"Technical Review Criteria" of the L-A. Harbor Enhancement Study Re-
view Criteria enclosed as guidance for reviewing the HEP's document.
1 As we understand it, the approach to the study plans for this
section was to collect data in the Los Angeles-Long Beach
Harbors during normal cannery operations with discharge to harbor waters
and subsequently compare them with data collected after discharge from the
canneries was diverted to the Terminal Island Treatment Plant. We
believe this is a reasonable approach, and if properly done, could
provide meaningful results.
However, we believe several erroneous assumptions and applications
of imperfect logic to data admitted to be inconclusive, have led to
erroneous conclusions. For instance-, DF&C fish block catch statistics
are used in the document to gauge populations. However, these dntn
only reflect catch and effort and not actual populations. For another
example, although many variables pertaining to population dynamics are
mentioned we do not believe the effects they may have are accurately
portrayed in several cases. Also, we believe the sampling techniques and
level of effort employed to collect data on population dynamics are
barely adequate for the task at hand.
In several cases, the report adnits that the data do not lend
themselves to statistical analysis. We believe the lack of statis-
bor development upon marine biota that might be found there whether
cannery discharges to the harbor continue are serious shortcomings
with the lack of a portrayal of future har-
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of this section. These deficiencies also reflect an incomplete state-
ment of qualifying factors- pertinent to any conclusions drawn from the
data. In short, the cause and effect relationships portrayed are based
largely on circumstantial evidence and coincidence with little re-
?liance on hard data having statistical significance.
Conwnent: "Erroneous assumptions, imperfect logic, inconclusive" data .... and
Mr. Kaneen looks forward to continuing dialogue with HEP?
What would DFG suggest in lieu of a non-existent, prior, long-term, ongoing
inshore fisheries trawl inventory program and an identical offshore program?
At the suggestion of SWQCB and DFG, we used the only data available to us to
attempt assessment of fluctuations in coastal populations over a large area.
DFG data are tabulated in a variety of ways, and the assistance in obtaining
data of several DFG people (Messrs Collins and Oliphant, Ms Wine) and Mr. Verna,
the bait dealer, was very much appreciated. Ms Cooksey and Ms Smith, former
DFG aides, carried out the dock surveys for HEP.
The reviewer decries the lack of statistical treatment but substitutes "belief"
(four statements on p. 10) and opinion on his part.
Comment: Sections IV and VIB of the HEP report do detail statistical treat-
ment of those data which could be so treated. The treatment presented is per-
haps different from what your reviewers are familiar with but are acceptable
means of data analysis. Stephens' fish data were treated with standard statis-
tical methods, as indicated.
What has the lack of certain statistical analyses to do with future harbor
development? We dealt with the latter in the Report to the Army Corps of
Engineers (AHF, 1975; 19.76) in the Master Environmental Setting (Port of Long
Beach, 1976), in the SOHJO EIR (SES-AHF-SI, 1976) and in the TITP Preliminary
Draft EIR, among others (all cited). Obviously the reviewer did not look at
any of the references. It should be noted that the Phase I TITP EIR deals only
with relocating the outfall pipe if the Los Angeles Main Channel dredging takes
place. The research report was not supposed to deal with any of these factors.
It was supposed to be a resource document and will be used as such. The DEIR
dealt properly with completion of the TITP project.
Taken by themselves, without qualifiers or assumptions, we believe
the data can just as readily'be interpreted as showing that since
cannery effluents were diverted from the harbor, populations have dis-
persed from an attractant focus to more norxaally random distributions.
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18
Comment: "... normally random distributions:. The data cannot be interpreted
to show that populations are dispersed in a random pattern. Fish and other
mobile organisms are not randomly distributed like thrown dice; they are dis-
tributed according to physical, chemical and biological preferences and/or
requirements. Since food is at least as important as habitat and protection
from predation, some fish will move long distances in search of food. Others
do not move any great distance and simply will not survive and reproduce if the
food source in their area disappears. By dealing with mean populations in the
Edison report the trends downward have been masked. Because this is such an
important point, we attach new graphs comparing the Edison data with the HEP
data, which show very similar downward trends.
Section IIA - FISH POPULATIONS IK LOS ANGELES-LONG BEACH ilAREORS
This section of the report portrays trawl data, creel census data and
catch statistics in an attempt to portray changes in fish populations. Our
analysis is as follows:
1. Page ID contains a statement that white croaker (Genyonemus
lineatus) are sold as "butterfish" at about $3 per pound. This
is probably an erroneous statement. White croakers are often
2/
sold in markets as either king! ish or tont cod — . The market
demand for this fish is limited to the Los Angeles area.
Current market price for this fish is about 70c per pound.
Butterfish on the other hand is the common name for
3/
Anoplopoma fimbria which is also known as sablefish — or black
2/
cod — . Although butterfish may sell for $3 per pound, it is
illegal to sell white croaker as butterfish.
Comment: It is not an erroneous statement. The quotation marks are purposeful;
so was the sale of croaker on the dock and the subsequent marketing of it under
2/ Gates, D.E. and H.W. Frey. 1974. Designated Common Names of Certain Marine
Organisms of California. In_ Fish Bulletin 161. Calif. Dept. Fish and
Game. 90 pp.
3/ California Administrative Code, Title 14, Section 103(2).
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the guise of "butterfish" It may be illegal, but it was being done. Contrary
to the statement, the common name "butterfish" is applied to several species
of the genus Peprilus on both the Atlantic and Pacific. The sable fish
(Anoplopoma fiiobria), also known as black cod, is not caught by dock anglers
but occurs only in deeper waters; the National Marine Fisheries Service has
several fisheries development documents which allude to these commercial
species.
2. On page IIA-4 Che statement is Bade that trends in fish popula-
tions in the harbor cannot be related to waste discharges. We
believe this to be an understateaent because the "populations"
cited are small parts of the larger whole which can r.ove freely
into and out of the harbor complex.
Comment: We are glad that the reviewer feels that the statement attributed to
us is an understatement. We feel that the reviewer makes a misstatement of
what was said in the text. A few years ago DFG thought the harbor Engraulis
was an endemic species and also that the harbor croakers never left home.
The statement on p. II A 4 is misquoted by DFG; it says "There is no practical
method for directly relating the fish populations to the wastes. However, ..."
Nevertheless we believe that
the trawl data reported are incomplete because they do not con-
tain data concerning such factors as salinity, water temperature
and other physical/chemical parameters. Cur experience has shown
us that these factors can influence trawl sampling.
However, even if we assume the parameters were recorded and
factored into the reported data, discussions of these data do not
(except in one or two cases) make comparisons between population
dynamics of a species within the harbor and the population dynamics
»
of the same species outside the harbor.
*Combs, Earl R.r Inc. 1978. A study to determine the export and domestic markets
for currently underutilized fish and shellfish. Prepared for U.S. Department of
Commerce under Contract No. M0-A01-78~00~4037. 376 p.
Anon. Fisheries Development Task Report. 1979. By NMFS, NOAA, Dept. of
Commerce. 106 p.
c ^
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20
Comment: Monthly records at 43 stations at 1 meter intervals through depth
were taken of temperature, salinity, DO, pH and turbidity, AS INDICATED IN THE
DOCUMENT. Records are used in interpreting all the biological data, as, for
example, in the multivariate analyses. It is not feasible to publish all raw
data, given the limits imposed by printing and mailing costs, and the limits
on EIR size imposed by CEQA. For the record, the heavy rainfall of December
1977 occurred after the two fish trawl dates.
No "population dynamics" studies were included, although fish measurements are
on file. Data were not presented because of the lack of comparable "offshore"
studies.
3. In the revised document changes in anchovy density in the harbor
( about 100 fold" decrease) are compared to changes in anchovy density
offshore ( four fold" decrease). Other than these references in
the text, no data are offered nor.is a definition of "offshore"
presented. If "offshore" carries its usual meaning (i.e., deep,
open ocean waters as opposed to near-coast shallow waters or
"inshore"), these comparisons are not surprising and not
necessarily reflective of changes in the harbor. This is because
data collected by the Department indicates that a reproductive
failure in 1976 caused a low population throughout the southern
California Bight. Reproductive failure could easily account for
a "100 fold" decrease in juvenile populations found in the harbor.
Comment: Offshore refers to the fact that the DFG reJeased the acoustical survey
data to the newspapers (Los Angeles Times, Mar. 79} on the anchovy stock off-
shore; it reflects the fact that inshore (in the harbor) there are now insuffi-
cient anchovy for a bait supply; it reflects the disappearance of Engraulis from
trawls where they used to occur in swarms; it reflects the fact that the egg
and larvae trawls "inside and outside" couldn't find eggs in season. The DFG
acoustical survey station data should be available to the DFG reviewer if he
is unaware of the locations sampled.
Comment: The statement by the DFG reviewer that "no data are offered" is not
true. Page 92, Table 18, of the HEP report is based on the records of The
California Department of Fish and Game, and this is indicated in the heading
for that table.
C"Y3
A
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21
Comment.- Note that the "reproductive failure" of 1976 was followed by a
modest recovery in 1977. The harbor used to b£ defended and protected from
development by DFG on the grounds that it was an essential nursery ground for
anchovies. One of the main reasons for the Corps of Engineers 1974-75 studies
(AHF, 1975; 1976) was the controversy over the threat to the nursery. Now
DFG would have it that there appears to be no connection between food and/or
habitat and "reproductive failure". On the contrary, there was the largest
egg and larvae census in recent history in 1975; the one-year class failed
to recruit, which is quite a different matter. The larvae didn't survive,
for unknown causes. But the enormous drop in the harbor coincident with loss
of particulate waste food and loss of phytoplankton, cannot but have affected
the harbor as a potential contributor to the "offshore" stock.
4.	Commercial parcyboat angler records are used to portray fish
populations in the vicinity of Los Angeles-Long Beach Harbors.
Again, DF&G catch statistics are intended to reflect only catch
and effort. It is a fundamental error to make population esti-
mates from these data because variables such as selectivity of
gear, number of fishermen, number of boats in the fleet, and
weather influence both catch and effort. Another source of
error results because the catch data are reported by the fisher-
men, or in some cases, by clerical personnel at markets and land-
ings where fish are unloaded. Fishermen are sometimes indifferent
and clerical personnel often uninformed about where fish were caught.
5.	The bait catch and creel census data are portrayed as
«
showing interesting, coincidental phenomena but, according to
the document, cannot be used to make statistical analysis nor, in
our opinion, can they be used to reach any sound conclusions of
•the type offered.	•
Comment: The creel census was done by two people who have been hired by the
DFG for conducting a creel census. The committee on the scope of work wanted
it done, imperfect as the creel census may be statistically, and coincidentally,
the results fitted in with other data. As indicated, these are considered to
be supporting data only. Attractive young ladies may have more luck than
uniformed wardens in getting people to talk about their catch, especially when
they are knowledgeable about fish species.
k.
c r>
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22
For example, a creel census requires careful documentation
of catch versus effort and complete enumeration of fish with
weight or length, or both. This normally is accomplished by
placing a person at a point of restricted entry, such as at the
foot of a pier, counting anglers' time of entry and exit, number
of anglers, number of fishing rods, and recording measurements of
fish caught. Because people are occasionally reluctant to show
their catch, uniformed personnel using prominent visual aids ate
used to overcome reluctance and to better assure completeness
of the data.
The creel census reported was conducted independently of
DF&G participation and census takers did not stay in one spot,
but moved throughout the harbor complex. Enumeration of hours
fished, number of fish caught and evaluation of the fishing ex-
perience were thus only as objective as fishermen were reliable.
Section 113 - MARINE ASSOCIATED AVIFAUNA OF OUTER LOS ANGELES-LONG BEACK
HARBORS IN 1978
We cannot agree with the conclusion that the bird survey results are
consistent with the concept that cessation of cannery effluent removed a
significant source of enrichment in the harbor food chain (Section ID).
Two sets of data were compared, but as the report points out, the two cannot
be compared directly. We. believe that without data from years between the
1973-74 survey and 1978 survey that trends cannot be accurately portrayed
for bird populations. Our staff expert believes avian populations in the
harbor area are healthy and stable. He also believes that because gulls
are no longer strongly attracted to Terminal Island outfalls, they are less
likely to prey on eggs of the California least tern (Sterna albifrons)
which nests nearby.
c?o
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23
Comment: DFG opionion: DFG doesn't agree with Dr. Power and his staff on the
bird data. Power has a Ph.D. in ornithology, is Director of the Santa Barbara
Museum of Natural History, and does surveys for the National Park Service and
other agencies. His conclusions are likely to be more valid than most. The
DFG "believes" and "believes" but gathers no harbor survey data; Power said
the data were not directly comparable but DFG distorts his words. In 1973-74
the surveys were done weekly and included the inner harbor. Only the outer
harbor data were compared, and data were averaged for count, a normal procedure.
Section IIC, IIP, HE - PHYTOPLANKTON PRIMARY PRODUCTIVITY IN OUTER LOS
ANGELES HARBOR 1976-78: CHANCES IN ZOOPLANKTON*
In" OUTER LOS ANGELES-LONG BEACH" "HARBORS: BEKIHIC
RESOURCES
Conclusions about fish, phytoplankton, zooplankton and benthic in-
fauna are controverted by a report prepared for the Southern California
Edison Company (SCE) concerning the effects of their Long Beach Generating
4/
Station, covering the period 1974 to 1978 — . The SCE report contains the
following conclusions, among others:
"The phytoplankton community in the harbor was characteristic
V
of nearshore environments common in southern California. Diatoms
were dominant throughout the year, with dinoflagellates con-
tributing appreciably to the population only in June 1974.
Analysis of all parameters and comparisons of trends...indicated
that long-term variability in phytoplankton populations in Long
Beach Harbor are due to natural yearly and seasonal fluctuations."
"The zooplankton community in Long Beach Harbor was typical
of southern California coastal waters. Primary components of the
cforraunity.. .were copepods and cirripedia nauplii. In 1976, a
cladoceran also became a dominant nieraber of the community. Data
suggest that major controlling factors of the zooplankton popula-
tion. .. included breeding strategies, food availability, predation,
and longevity of individual species within the plankton...."
j4/ Environmental Quality Analysts and Marine Biological Consultants, Inc.
1978. Marine Monitoring Studies Long Eeach Generating Station, final
^	report 1974-1978.
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24
"The infaunal community was characterized by Inner to Outer
I
Harbor gradients in physical and biological parameters which act
to divide the species complexes. Although species turnover rates
were high harborvide, dominant species demonstrated relatively
consistent patterns of distribution. The influence of natural
variation was reflected in the wide fluctuation in population
density for most of the dominant species. Overall, the diversity
and abundance of organisms in the faunal assemblages were typical
of those In comparable restricted harbor areas of the California
coast...H
"Nekton studies showed that Long Beach Harbor supports a
large, healthy, stable, and diverse population of fishes that are
common in nearshore southern California waters. With Binor
rearrangements in numerical abundance rankings, the same species
maintain a dominant position throughout the study period. The
species were: white croaker, northern anchovy, queenfish,
California tonguefish, white surfperch, and shiner surfperch"
(emphasis added).
Commert: Criticism of Section IIC, IID and IIE of the HEP report relies almost
exclusively on quotes from the 1978 EQA-MBC report prepared for the Southern
California Edison Company. That report addresses results of environmental
monitoring carried out to assess changes in the environment as a result of a
thermal discharge in a very limited area of the harbors, inner Long Beach Harbor.
1.	The DFC reviewer failed to note that different time periods were covered by
the two reports. EQA-MBC completed their field work in early 1978. The Edison
report was not even available when we sent the first draft of our volume to EPA
in Washington in February, 1979. Our HEP work continued through 1978, the
year in which great change was noted. Indeed, the coincidence of events we
have observed, relating waste discharge to biological fluctuations during that
year, occurred after the end of the EQA-MBC study.
2.	Different areas of the harbor were sampled in the two studies. The EQA-MBC
study for Edison was almost exclusively limited to the Middle and Inner Harbor
areas of the Port of Long Beach, where the principal feature being considered
was the effect of a thermal discharge on the marine environment. These areas
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25
are different in habitat and flushing and tend to be somewhat isolated by the
pattern of the currents from the outer Harbor areas studied by HEP. This
observation is supported by the work of EQA-MBC and by independent studies by
the U.S. Army Engineer Waterways Experiment Station, Vicksburg, Miss. (McAnally,
1975 referended in HEP).
Outer Los Angeles Harbor is where the major and most immediate effects of the
waste discharges from the TJTP and the canneries occurred. We do have a report
in press concerning the entire harbor in 1978, updating the 1973-74 studies
for the Corps of Engineers by HEP. (EXCERPT OF RELEVANT DATA APPENDED) .
3. A limited amount of sampling by EQA-MBC was carried out near some of our
stations. Where the sampling coincided in time and location there is sub-
stantial agreement between their data and HEP data. If the two Edison trawl
stations that are located in the outer harbor are compared with the HEP trawl
data from that area, the data are very similar where they overlap in time. The
remainder of the trawl stations (nekton) of the Edison survey are in the inner
and middle Long Beach Channel/ only the two can be even considered as evaluating
the outer harbor.
HEP phytopiankton and zooplankton results are not controverted by Edison data.
They stopped their survey in March, 1978. All the Edison plankton stations
are in the inner and middle Long Beach Channels and not a single sampling sta-
tion is relevant to the TITP study.
Section IIF - FISH EGG AND LARVAE CENSUS
Section IIIA through E - MICROBIOLOGICAL CYCLING OF NUTRIENT
Section IVASB - INTERACTION OF PHYSICAL AND BIOLOGICAL PARAMETER
Section VA - PHYTOPLANKTON GROWTH AND STIMULATION IN THE TERMINAL ISLAND
TREATMENT PLANT SECONDARY WASTE PLUME
Section VP - GROWTH AND STIMULATION OF INVERTEBRATES IN THE WASTE PLUME
We have no criticism to offer concerning these five sections of
the HEP's report except to note that they reaffirm what has been generally
known for some time, namely that populations fluctuate with food supplies or
oc.her variables. This might be an important consideration if nutrients
were limiting environmental factors in the. harbor system, but this is
apparently not the case.
Within this context, however, we believe it is useful and important
to make a distinction between enhancement and the concept of stimulation
used in the HEP report. Enhancement, from our view, means augmentation or
C 3
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26
improvement and is fully compatible with the idea of uninterrupted protec-
tion of beneficial uses. On the other hand we find that to stimulate means
~»	5 /
to rouse to activity or heightened action as by spurring or goading...." — .
The synotiya offered at this reference is to provoke. We think this defini-
tion is clearly at odds with the idea of uninterrupted protection of
beneficial uses because although "stimulate" often has positive connotations,
it is an ambiguous term. It can also carry a negative meaning in the sense
that excessive stimulation in a given case could cause a reversal of any
benefits that may have accrued earlier.
The distinction we have portrayed between enhancement on one hand
and stimulation on the other is for us a fundamental reason why demon-
strations of enhancement must be beyond doubt. In our opinion, it is
logically false to equate biostimulation with bioenhancement as the
HEP report seems to do. We must point out that in the sections listed
above, we see ample demonstration of the stimulatory nature of efflu-
ents from canneries and the waste treatment plant at Terminal Island.
However, the demonstration of enhancement is not clear.
Section VB - TERMINAL ISLAND TREATMENT PLANT SECONDARY WASTE BIOASSAYS
The results of this study suggest that test organisms of hardy species
(Pundulus: Neanthes), that occur naturally in waters unsuitable for many
other aquatic species, will survive an increased concentration of dissolved
organic matter if sufficient oxygen is supplied.
5/ Norris, W. (ed.) 1973. The American Heritage Dictionary of the English
Language. American Heritage Pub. Co. and Haughton Mifflin Co. Boston;
New York; Atlanta; Ceneva, 111.; Dallas; Palos Alto
f
A
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27
The results also suggest that more sensitive species (Engraulis) may
be greatly affected by cannery or treatment plant discharge. We believe
this study would have benefitted if tests involving anchovies (Engraulis
aordax) were repeated, (with different effluent concentrations, if appro-
priate) and I-C^q values calculated.
This is of particular interest because a strong claim is made else-
where in the document that waste effluents from the canneries and Terminal
Island Treatment Plant have enhancing effects upon larval anchovies such
as were used in this test series.
Comment: HEP couldn't continue to use Engraulis; they disappeared, as has been
documented earlier. HEP followed the EPA/Corps of Engineers manual of bioassay
procedure where appropriate,and used representative harbor animals. The DFC
reviewer has not understood the statistical analyses apparently or he would
not have made these statements. With regard to concentrations, the cannery
wastes formerly underwent a natural 50% dilution within 100 yards. The dilu-
tions of the enhancement zone were naturally lower. That does not negate the
concept of enhancement. It does demonstrate the present inappropriateness
of the regulations.
Section VC - CANNERY WASTE AS A FOOD FOR ANCHOVIES
The test regime described in this section of the document is different
enough from that of the previous section that methods should have been
more fully described. For instance, the methods used to collect test fish
and the length of time fish were held before testing are important to
know before meaningful review of the data can be made. Nevertheless, we
offer two comments concerning the results as presented.
First, if the inference is correct that cannery waste causes growth
as great as or greater than "trout chow'', (presumably a commercial product)
then it seems to us that a potentially marketable fish food source is
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28
being thrown away, and perhaps should be reclaimed rather than discharged
to harbor waters or to the sanitary sewer system.
CommentIt is a great surprise to HEP that the DFG as guardians of the natural
environment should recommend a commercial use of the resource over the good
health of the free-living marine populations. The fact that the wet sludge
would have to be dried (an Air Quality problem and an Energy problem), and
sterilized for general sale may make it an inappropriate commercial fish food.
That is a question of economics, and does not affect the question of whether
fish thrive on the semi-solid sludge.
On the other hand, although no data are offered, the document claims
that mortalities were "significantly fewer in the sludge fed group" (page
497), but on page 498, the document says "mortality, although high,
averaged 41% and was not significant from tank to tank." These opposing
statements are difficult to interpret without data.
Comment: "Mortality differences", as stated, were not significant between
replicate tanks. The phrase "from tank to tank" was taken out of context.
As discussed in the review it indicates that the reviewer did not relate the
words to those preceding ther. and following them in the same sentences. We
suggest that the reviewer consider these statements in the context of their
presentation.
However, even dis-
counting some mortality resulting from experimented conditions, the report
as written indicates that cannery sludge discharged directly to harbor
waters may adversely affect a significant part of fish populations in the
harbor even though it "enhances" the rest. If true the net benefit to
the population is apparently nil and demonstrates, in our opinion, why full
and uninterrupted protection of beneficial uses in estuaries and embay-
ments is preferable to an averaging concept.
Comment: The report as written does not indicate that cannery sludge discharged
directly to harbor waters may adversely affect a "significant" part of fish pop-
ulations, as claimed by the reviewer. No such statement was made or implied, nor
can such a statement be supported by the data. How did DFG establish a signifi-
cance test for that statement?
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29-
Irt summary, we have always welcomed criticism and encouraged cooperative parti-
cipation in the hopes of making a contribution'to the environmental health of
our n&rine neighborhood. The attempts of DFG to discredit some 38 experienced
professional marine researchers and an equal number of technical people does
little to distinguish the contribution of DFG to enhancing marine resources.
HEP has obtained input and funds from a wide variety of sources and scopes to
build the only long term harborwide baseline in existence. Genuine scientific
discussion is needed, along with continuing efforts to develop new concepts
and regulatory strategies. The traditional approaches around the country have
not been uniformly optimal, to say the least.
There are several issues involved in the critique submitted by DFG. First, HEP
has always welcomed participation of agencies and colleagues in planning and
implementing studies. Advice can only be accepted so long as it fits within
the purposes and scope of work of funded investigations. Personnel from
agencies and other institutions have often participated cooperatively in
ongoing field and laboratory studies. Second, HEP welcomes constructive
criticism and discussion of results. Since no study can address every pos-
sible aspect of every condition or circumstance encountered in the environ-
ment, discussion of causative factors and interpretation of results is
essential. Third, attempts by DFG to discredit some 38 professional marine
researchers and an equal number of technical staff from several respected
institutions is counter-productive and does little to distinguish the DFG
contribution to enhancing marine resources. Fourth, the efforts of HEP
colleagues to build a harborwide data base by linking their own contracts and
grants to common scopes where possible have provided a baseline that would not
otherwise have existed, even though there are, unfortunately, some gaps in it.
Finally, there are urgent needs for new concepts and regulatory strategies
to improve those areas of environmental control which have not been optimized
under traditional approaches. HEP believes that the interactions in the
marine environment of domestic wastes and non-toxic, nutrient-source wastes
such as fish cannery effluent, have not been managed satisfactorily under
present regulatory processes. HEP has sought to ameliorate the conflicts
of uses and goals in the urban harbors while enhancing the marine life therein.
err
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ATE OF CALIFORNIA—RESOURCES AGENCY
•EPARTMENT OF FISH AND GAME
ARINE RESOURCES REGION
50 Golden Shore
yong Beach, CA 90802
;213) 590-5117
24 May 1979
EDMUND G. BROWN JR., Gevmor
Mr. Jeffery D. Denit, Chief
Food Industry Branch (WH-552)
United States Environmental Protection Agency
Washington, D. C. 20460
Dear Mr. Denit:
Corrections to Review of Report Submitted to EPA
by Dr. Dorothy Soule of the Harbors Environmental Project
In the review accompanying the letter we sent you dated 22 May 1979,
a footnote was inadvertently omitted from page 11. The footnote
should have read as follows:
2/ Gates, D. E. and H. W. Frey. 1974. Designated Common Names
of Certain Marine Organisms of California. In Fish Bulletin
161. Calif. Dept. Fish and Game. 90 pp.
In addition, the following errors need correction (corrections under-
lined) :
page 1, second paragraph, next-to-last sentence: ... the
enhancement issues comprise an important perspective that
must be understood while these issues are debated.
page 8, 6th line: Non contact water recreation ...
page 8, 16th line: and perceived^benefits to the biota ...
page 10, second paragraph, second sentence: We believe the
lack of statistical analyses in concert with the lack of a
portrayal of the impacts future harbor development will have
upon marine biota ...
page 15, first line: maintained a dominant position ...
page 15, sixth line: MICROBIOLOGICAL CYCLING OF NUTRIENTS
page 15, seventh line: INTERACTION OF PHYSICAL AND BIOLOGICAL
PARAMETERS^
C' 3
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Mr. Jeffery D. Denit, Chief
-2-
24 May 1979
We hope, our errors have not caused you inconvenience as you read our
analysis, and that these corrections resolve any questions you may have
had. If you have additional questions about our review, please call
Kuuci L	IVQUCCU
Regional Manager
cc: Dr. D. Soule, HEP
SWRCB - Div. Water Quality - Howard Wright
NMFS, Terminal Island (Jim Slawson)
USFWS (Jack Fancher)
RWQCB #4 (Dr. Lewis Schinazi)
EPA-SFO (Terry Brubaker)
us at (213) 590-5136.
Sincerely,
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UNIVERSITY OF SOUTHERN CALIFORNIA
1
Institute for Marine mid Coaatal Studies
UNIVERSITY PARK ' LOS ANGELES, CALIFORNIA 9OOO7
imiLSJ
„ : fc t f k f --s i
Harbors Environmental Project#
Allan Hancock Foundation, Rin 139
July 3, 1979
Mr. Robert G. Kaneen
Regional Manager
Department of Fish and Game
Marine Resources Region
350 Golden Shore
Long Beach, CA 9 0802
Dear Mr. Kaneen:
Thank you for your courtesy in sending us a copy of your letter
to Mr. Denit and of the review by your staff of our report
entitled:
We have read with considerable interest the critique of our
work and because of the many issues raised, we have inserted
our comments in a copy of your critique (see attachment).
In the closing paragraph of your letter to Mr. Denit you
indicate that you "look forward to a continuing dialogue with
Dr. Soule and her colleagues." We regret that this was not
done prior to distribution of the review.
We have never claimed that our research program was beyond
improvement. This is why we welcomed observers for all of
our research activities and solicited comment from knowledge-
able professionals such as Dr. Howard Wright of the State
Water Resources Board, Dr. Lewis Schinazi of the Los Angeles
Regional Water Quality Control Board and Dr. Christopher
Stevens, then the environmental scientist for the City of
Los Angeles Department of. Engineering. We also welcome
Ecological Changes in Outer Los Angeles-Long
Beach Harbors Following Initiation of Secondary
Waste Treatment and Cessation of Fish Cannery
Waste Effluent.
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Mr. Robert G. Kaneen
July 3, 1979
Page 2
unbiased professional review of our reports; we have sought
reviews in the past and will continue to do so. Some of the
changes in methodology over the eight year period were intro-
duced at the suggestion of others. One must recognize that our
research was not a single project under one long-term contract.
Therefore, in putting together results of HEP contracts, grants,
and graduate student research, some changes in scope do occur.
It still is better than the much favored literature survey of
data from non-comparable sites, periods and methods.
Unfortunately, we find that the DFG critique contains misrepre-
sentations and distortions of statements in our report. For
example, on page 11 of the critique, item 2 references a
statement in our report but alters the meaning and intent of
the statement. The critique displays some surprising over-
sights - e.g., page 10, paragraph 2 of the review decries
the lack of statistical analysis. This suggests that the
reviewer did not see or did not understand Section IV of our
report which presents such analyses, or Section VI with 50
pages of advanced methodology for programs originated by
EEP and now in use by Edison and many consultants.
The DFG reviewer relies on his opinion unsupported by any
substantive documentation to refute our findings which are
based on data from our studies and from other independent
studies (e.<£. , on page 10 of the review the word "believe" is
used four times without any indication of the basis of belief).
In closing, we again would like to express our appreciation
of the courtesy extended by you in sending us a copy of this
critique of our work.
Sincerely,
Dorothy F. Soule, Ph.D.
Director
Mikihiko Oguri, M.S.
Associate Director
Harbors Environmental Projects
Attachment
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Ecological Changes in outer Los Angeles-long Beach Harbors since the Initiatior
of Secondary Waste Treatment and Cessation of fish Cannery Waste Effluent at
Termina1 Island, California. Harbor Environmental Projects, University of
Southern California. February, 1979.
Reviewer and review date: Richard C. Swartz, 28 March 1979
GENERAL COMMENTS
Bioenhancement is a useful concept for coastal zone management. A good
example is the creation of artificial reefs constructed from non-toxic
materials. When placed in habitats with little physical heterogeneity, e.g.
sandy areas of the continental shelf, such reefs attract fishes and other
marine organisms. Such effects are desirable from human and ecological
perspectives.
The introduction of nutrients through wastewater point sources into the
marine ecosystem has the potential for both detrimental and beneficial effects
Adverse impacts are very likely if the effluent contains chern'cal pollutants
or other factors that exert toxic effects on marine organisms. Therefore the
threshold bioassay criterion of the California Bays and Estuaries Policy is
essential in the application of the bioenhancement concept. Earlier Harbor
Environmental Projects (HEP) reports have indicated that the L. A. seafood
cannery effluents are acutely toxic to ecologically important species such as
the cope pod', Acartui lonca and anbryos of the anchovy, Enjixiuliv	at
concentrations well below the 90':1 level included in the Bays and Estuaries
Policy criterion. The present draft HEP report does not include the section
entitled "Bioassay of Invertebrates and Fish". This section must be submitted
and reviewed before any further regulatory decisions can be made about
bioenhancenent in L. A. Harbor.
If wastewater effluents are not toxic, biological conditions have to be
assessed to determine whether enhancement is evident. The qualitative
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biological criteria for enhancement given in the HEP report may be useful for
guidance, but I doubt that any list of biological parameters could be
universally applicable, especially in a quantitative fashion. Biological
impact assessments must be based on site specific considerations.
1 disagree with the HEP suggestion that biological conditions should be
integrated in space to determine if there is an "overall" enhancement
(P1A5). A claim of enhancanent should be denied if there are adverse impacts
at any point beyond the immediate vicinity of a point source.
I agree with the statement (PIA10) that no single criterion should be
considered sufficient to qualify as bioenhancement. However, an adverse
impact evident from any one criterion should be sufficient to deny a claim
of bioenhancement.
Criteria should not be based on diversity indices, especially those
derived from information theory (PIA12). These indices are not unequivocal
indicators of community "health". They may be useful in assessing ecological
conditions, but only when their patterns are interpreted in terms of other
aspects of community and population dynamics.
I agree with the report's conclusion that biomass, by itself, is not a
very good biological criterion (PIA14). If a diverse, indigenous biota is
replaced by a more abundant assanblage of a few opportunistic species, the
increased standing crop should not be considered bioenhancement. This is not
consistent with the report's earlier listing of a criterion based on "the
presence of living biomass, above that which would occur in the absence of
the discharge". (PIA10).
I do not believe that the HEP criteria adequately addresses population
characteristics such as disease incidence, size frequency distribution,
reproductive potential etc. Certainly the condition or quality of populations
must be examined in addition to their relative abundance.
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SPECIFIC COWENTS
• METHODS:
Survey and experimental methods are inadequately described in several
sections of the report, especially part II E which deals with the benthos.
Table 1 in section I B indicates that the benthic fauna was sampled quarterly
with a "Cambell grab or Reinecke box cover, 0.5 mm screc.n". No additional
details are given in section II E. The report does not indicate the size of
the grabs. SCCWRP has shown that different grabs give different results.
The report does not indicate which samples were taken with the Cambell and
which with the Reinecke. The number of replicates at each station is not
given. A statistical analysis of the benthic data is not presented. The
data is restricted to graphs of temporal patterns in areal species richness
and density of the benthic macrofauna. Only the dominant taxa at a few of the
stations are identified in the report. It is very difficult tc make any
conclusions about ecological changes on the basis of such a limited and
qualitative benthic analysis.
The methods of section V C "Cannery Waste as a Food for Anchovies" do not
meet one of the most basic requirements of bioassays: low control mortality.
The EPA Ocean Disposal Bioassay Procedures state that "A test is not acceptabl*
if more than 10 percent of the control animals die". In the anchovy growth
experiment, 41 % of the control specimens died.
EXECUTIVE SUMMARY:
The executive sunnary identifies certain ecological changes that may
have occurred in L. A. Harbor between 1973 and 1978. In several instances the
summary and report fail to present convincing evidence that these changes
are due specifically to the conversion of cannery effluents to secondary
treatment. Some beneficial effects of that conversion are ignored.
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The conclusion that bird populations are down 40?; is based on a comparison
• of two major surveys made in 1973-4 and 1978. Data for the intervening four
years, which might show that the decline is temporally related to the treatment
E
conversion, are not available. Furthermore, the report states that the results
of the 1973-4 and 1978 surveys are not directly comparable (PIIB7).
Conclusions about declines in fish and benthic populations are based on
more complete temporal surveys since 1972, but many of the trends began long
before the conversion to secondary treatment and may have been caused by a
variety of human and natural factors. Dr. John Stephens' report on fishes
concludes, "Certainly, there is no indication that cessation of discharge
has been beneficial to fishes, but because of variations in background levels
of populations it is impossible at this time to state that there has been a
detrimental effect" (P11 A3). He also notes that decreases in fish abundance
may be occurring throughout the southern California bight.
There is evidence of beneficial effects after conversion for benthic and
zooplankton communities near the outfalls. At station A7 benthic species
diversity and abundance increased in 1978 after conversion, to levels equal
to or greater than data collected from 1971 to 1977. The pollution tolerant
polychaete Capiiclti capitals was replaced as the dominant species by
calij'ormumr.is. Mo 11 uscans and crustaceans increased in abundance.
According to Reish's classification the benthos changed from that typical of
a polluted zone to that of a semi-healthy zone.
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SECTION II F Fish Egg and Larvae Surveys
Ichthyoplankton species diversity, density of fish eggs, and density of
fish larvae were consistently greater in 1978 (after conversion to secondary
treatment of cannery wastes) than in 1974. The differences in d-ensity were
substantial; sometimes greater than two orders of magnitude. The HEP report
suggests that these differences are "probably due to reduced predation by adult
fish and to more efficient sampling methods". This report does not adequately
consider the more obvious possibility that the conversion to secondary treatment
may have eliminated a significant stress on the ichthyoplankton. That possibility
is supported by an earlier HEP report (1976, Part 12 - Marine Studies of San Pedro)
in which Brewer reported bioassay results on the toxicity of the original cannery
and sewage effluents and concluded that "the cannery outfalls exert an overriding
influence of the toxicity of the effluent field to E. mordax (anchovy) embryos
and larvae".
SECTION IV B Weighted Discriminant Analysis of Benthic Data
The restriction of the benthic data set to polychaetes and molluscs may
create an initial bias in this analysis. Amphipods and echinoderms are much more
sensitive to wastewater effluents and their exclusion may obfuscate spatial
patterns around the outfalls.
This section suffers from a lack of a synthesis or discussion of the data.
The text lists about 20 factors that may have affected the benthos. It is very
difficult to identify causal relationships in the midst of so many correlations.
The spatial patterns of the species groups show that the benthos at the
stations nearest the outfalls are distinctly different from the remainder of the
collections. A comparison of Figures 1 and 29 shows that this pattern did not
change greatly during the survey. However in 1978 the diversity and density of
the benthos at the stations nearest the outfalls increased substantially.
SECTION V B Terminal Island Treatment Plant Secondary Haste Bioassays
The data in this section do not support the conclusion of the Executive
Summary that "In bioassay/toxicity tests there was no evidence that the secondary
TITP effluent was toxic at any concentration". All of the Emerita test specimens
died in the 100% effluent concentration treatment in test 2. In tests 1, 2, 3,
and 4 the control survival of anchovy embryos v/as always greater than in the 562,
75%, and 100% effluent concentrations and in the treatment consisting of receiving
water from the outfall boil. The HEP report suggests that this is a methodological
artifact. In a supplemental bioassay using a different exposure method, there
was high anchovy survival in all test concentrations. However, the supplemental
test was conducted at a mean temperature substantially below other test
temperatures. In fact there is a good relationship between mean survival in all
treatments and mean test temperature:
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Test
Number
Mean
Temperature
Mean
Survival
Suppl.
2
4
3
5
12.3
15.6
17.1
17.9
19.5
20.8
94.2
63.4
19.6
10.3
12.9
? No data given
Brewer (Fish. Bull. 74:433) has shown anchovy embryos develop normally at
temperatures between 11.5 and 27.0°C. The temperature-survival correlation in
the HEP results therefore suggests the possibility of a synergestic relationship
between temperature and toxicity. This is supported by earlier HEP results that
the LC50's for the cannery effluents released from the old Starkist and Way Street
outfalls were 17-18%.
SECTION V D Growth and Stimulation of Invertebrates in the Waste Plume
Mussel Growth
This section reports a statistically significant, but small difference
in growth rate of mussels placed at increasing distances from the TITP
outfall. Although mussels at the outfall had the fastest growth, mussels
at station A7, 550 m from the outfall, had the slowest growth rate. Thus,
if the effluent enhances growth, that effect is not widely distributed in
the harbor. Other factors such as temperature may also have affected
growth rates inside the harbor.
Biostimulation of Invertebrates
The data on biomass and diversity of settling organisms do not show
any spatial patterns that can be attributed to the outfall. The data do
not support the conclusion that the effluent plume "is providing nutrients
to a food chain which enhances growth".
Flow-Through Studies
These results are also inconclusive. There were no significant
differences in the growth of Mercenaria exposed to seawater with and
without wastes. There were no significant differences in the growth of
Fundulus that were starved, fed trout chow, or fed cannery wastes. These
results certainly do not support the conclusion that the pre-DAF waste
"could have a positive biostimulatory effect on some marine species".

A
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Robert Schaeffer (WH552)
Director of Effluent Guidelines Division
Environmental Protection Agency
401 M Street Southwest
Washington, DC 20460	f f?A
angei^A?;*
Re: Comments on ECOLOGICAL CHANGES IN OUTER LOS ANGEL'fc^A/JjJG, BEACH
HARBORS FOLLOWING INITIATION OF SECONDARY WASTE TRE^fNT/AND
CESSATION OF FISH CANNERY WASTE EFFLUENT. Marine Studiei'of
San Pedro Bay, California Part 16^Aprilyti^

Dear Mr. Schaeffer:
It is recognized that the Environmental Protection AgtjfTcy (EPA) is required
by Sec. 74 of the Clean Water Act of 1977 to conduct an investigation of the
ecological effects of seafood processing waste discharges to the marine
environment. It is also known that a portion of EPA's effort in this
investigation relates to Los Angeles Harbor, where several large tuna cair-
neries have traditionally discharged their wastewaters. The seafood industry
has advanced the concept of 'biological enhancement' of receiving waters by
seafood wastes. On the basis of the subject document prepared by Dr. D. Soule,
Harbors Environmental Projects (HEP), USC, they contend that their previous
waste discharges expanded the scope and magnitude of the marine food web and
converted the soft bottom community to a more productive environment. The
L.A. City Department of Public Works is also contending that sewage discharges
from the TITP have enhanced the harbor marine ecosystem.
Because the Fish and Wildlife Service is concerned for the important biological
resources of Los Angeles Harbor, we are aiding the EPA in this effort to evalu-
r
ate "bioenhancement", as much as manpower and funding allow. However, due to
.s''
the latter constraints, our review must be more cursory than the significant
issues dealt wi.th in the. subject document warrant.
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We offer the following informal comments on the subject document.
The premise under discussion throughout the document is that the discharge
of organic fish processing wastes and primary sewage into the harbor waters
had "bioenchanced" the harbor ecosystem. This premise is tested by compar-
ing data gathered while such discharges were occurring with data gathered
after initiation of secondary waste treatment and cessation of direct fish
cannery discharges. Though much of the sampling procedure and station
locations originated with different projects ^nJJtoonsors, the continued use
A'-*
of them to deal with the "bioenhancement" questiori*3^Sf seem inappropri-
ate. However, it is possible that a more tailored experimental design would
have yielded more definitive results. Further, the actual questions asked by
the investigators seem to have evolved well after much of the actual conduct
of the investigation. The sampling that was resumed in 1977 was somewhat
reduced in scope from previous sampling efforts and often employed dissimilar
techniques which rendered direct comparison impracticable. Therefore, none
of the 'before and after' sampling is statistically tested for significant
differences.
In the introduction, "Bioenhancement: Can This Concept Be Defined and Measured?",
many criteria are discussed which indeed should be used for determining bioen-
hancement (i.e. species diversity, evenness, richness, biomass, essential food
web species, species of commercial/recreational value, rare and endangered
species). Missing from that discussion is the investigator's expectation of
how "bioenhancement" would be manifested in each of those criteria. Neither
the author's nor any regulatory agencies definition of "bioenhancement" is
presented.
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Absent from virtually all summary statements are reports of the adverse
impacts associated with the cannery discharges and primary sewage discharges.
For example, Section II F, Fish Egg and Larvae Surveys p. 211, fails to
mention that during the treatment plant failure of July and August of 1978,
the density of fish larvae in the harbor plummeted. The high concentrations
of ammonia and/or chlorine probably proved fatal to larval fish. The overall
increased density of larval fish	by the 1978 studies could, in part,
be due to the reduction of toxic material concentrations in the harbor since
the 1974 study. The "zone of inhibition" ^^^^^«round the outfalls, when
they were discharging cannery wastes and primary sesa^7 receives scant atten-
tion. The vicinity of the outfalls suffered repeated episodes of high BOD
loads and reduced dissolved oxygen levels (see attachments).
Portions of the study lack a control site (e.g. bird survey, trawl survey). The
selection of an appropriate contxol area would seem difficult, however, con-
sidering the magnitude of the study area (outeT Los Angeles and Long Beach
Harbors) and the different questions being asked when the study first began.
The eastern end of the harbors, east of Pier J and south of Belmont Shores
might have sufficed as a control area, with some qualifying factors.
In order to compensate for the inadequacy of control area comparisons, the
authors could have incorporated the findings of other pertinent major studies.
It is likely that discussion of the results of other research programs in
nearby areas would have aided in a fuller accounting of biological trends of
the area and the responsible environmental factors. For example, the Southern
3
C ^9
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California Coastal Water Research Project has a primary objective of under-
standing the ecology of the open coastal waters of southern California,
especially the effects of the discharge of municipal wastewaters on sealife.
That project is as old as Harbors Environmental Projects; much of the informa-
tion generated during SCCh'RP's ongoing research could have been used for
comparison to HEP's information. Similarly, Southern California Edison had
funded a thorough marine biological study, jointly conducted by Environmental
Quality Analysts and Marine Biological Consultants, Inc., in Long Beach Harbor
spanning the years 1974 to 1978. SamplingSg^y^ftnd methodologies are dif-
ferent, thus precluding direct comparison, but the^jeJ^ral trends are of some
import. The extensive trawl and gill net sampling, mostly within Inner Long
Beach Harbor, indicates increasing fish population densities since 1976.
Because of the circumstances and conduct of the study which preclude statis-
tical tests, some of the conclusions of the study are based upon subjective
interpretation of the data, recognition of coincidental actions that may or
may not actually be related, and circumstance rather than scientific proof.
Often, the summary inferences are not the only on supported by the data.
Much is made of Trawl 13 during the July 1978 sample period. This trawl station
is nearest the TITP outfall; this sample occurred during the treatment plant
failure which resulted in the discharge of high levels of BOD and suspended
solids into the harbor after a period of some months without such discharges.
The unusual feature of this trawl catch is the nearly 800 individual white
croakei^, -frhilc -otfie*1 trawls taken nearby during the same sample had more
normal catches. This is advanced as support for bioenhancement.
4
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A fuller discussion of this episode should have included the following: A
trawls at the same site can result in an order of magnitude of difference
in catch. Kater clarity can be a factor in avoidance of the trawl net by
fishes. It is entirely possible that the turbid waters along the trawl path,
combined with a surface condition (lower salinity and dissolved oxygen
generated by the discharges which caused the fish to concentrate nearer the
bottom), created a circumstance that alftWed the abundant trawl catch. The
environmental conditions prevailing at th(	this trawl are unknown.
Whatever the conditions, there were more fish caugnt near the outfall than
farther from it. At best, the conclusion should be that the white croaker
were attracted to the area by something, probably some component of the
material discharged during the treatment plant failure. Though an organism
may be attracted to something does not necessarily indicate that the organism
will benefit from the attraction. Also the croaker had to already be in
evidence in order for them to appear at the outfall in July to be "enhanced".
That is, the harbor ecosystem was supporting them in the months prior to the
treatment plant blow out. The seasonal trend of fish abundance inside the
harbor, with or without cannery or treatment plant discharges, has been peak
abundances in the summer with the lowest abundances in the winter. This hap-
pens to coincide with the pertinent discharge events. Therefore, it is actually
unknown just what role was played by terminating the direct cannery discharges
in the winter since fish abundances decline to a seasonal low. Similarly,
the treatment plant failure occurred during the seasonal peak in fish abun-
dances. A synopsis of each chapter and the support it provides to the bio-
enhancement issue follows.
single trawl sample has considerable inherent variability. Day versus night
5
r'?.
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Chapter I A, B, § C - Introduction
An adequate discussion of the issues is produced along with quantifiable
parameters for measuring bioenhancement. As mentioned earlier, there is no
statement by the author or any other party of what must be proven to est-
ablish bioenhancement.
Chapter II A, Fish
Abundances of two species white croaker and northern anchovfy'account for
the large declines noted. No significant changes in species diversity is
established. (It should be noted that	purveys sample only a portion
of the total fish community. Larger and moA^ptjfit fishes, eg. sharks,
bonito, barracuda, corbina, are able to avoid the net. Trawls also miss
fish which usually associate with structures of the surface, eg. blennies,
topsmelt/^and jacksmelt, some surfperches.) Because the harbors are an open
system and an integral component of the larger offshore ecosystem, the
relationship of demersal fish populations to the cannery and treatment
plant discharge is not assessable with this type of study. However, there
is an indication that the white croaker population may have been stimulated
by harbor discharges. (Benthic feeding habit and apparent tolerence of low
dissolved oxygen levels accounts for much of its success in the more stressful
harbor environment of several years ago.)
Chapter II, B - Birds
The notable changes are fall and winter declines of several gull species
and surf scoters. Natural variability in the movements and occurence of
migratory birds can account for these changes. Some gull species, with a
scavenging habit, may have abandonned the cannery outfall as a feeding site
once the gurry was turned off. However, there is no support for the bio-
C",3	^
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enhancement contention in this chapter. Actually the two Federally listed
endangered species found in the harbor, the California least tern and the
brown pelican, may encounter less competition, and the tern less egg pre-
dstionjwith fewer gulls around. Not all species decreased. Some have
increased considerable, eg. western grebe, brown pelican, double-crested
cormorant. The decline of shore feeding species, eg. sanderling, turn-
stones, willet, sandpipers, cfould be due to elimination or disruption of
virtually all feeding habitat. Unmodified shoreline and undisturbed sandy
Co re-
beach has become exceedingly «ra-b&» in the harbor.
Chapter II ,C - Phytoplankton
No appreciable changes noted. No supppr^/for bioenhancement.
Chapter II,D - Zooplankton	* £
Changes noted are within the bounds of natural variability. No support for
bioenhancement.
Chapter II, E - Benthos
Bottom dwelling organisms are quite useful in tracking water quality changes
due to their short generation times and relatively sedentary life style.
While regional oceanographic conditions influence the harbor benthos, the
benthos is probably more significantly influenced by local conditions.
The results produced in this chapter seem to demonstrate the classic response
of a biological community to an organic pollutant. A few tolerant species
(i.e. Capitella capitata, a polychaefce worm) are able to grow abundantly in
high concentrations of the pollutant. As the pollutant is diluted other
species can live. At some distance from the source biomass reaches a peak
7
c
-------
and diversity levels off. If part of biocnhai\cemcnt is construed as
maximizing biomass and diversity then there is probably such a zone around
the outfalls. As clearly indicated in the study though, there is a zone
of inhibition associated with the outfall which must be weighed against
the zone of enhancement.
Chapter II, F - Fish Egg and Larvae
Overall increase in fish larvae is attributable to many causes, eg. different
sampling methods or reduced mortality in the harbor, but an adverse influence
of the cannery and treatment plant discharge is also possible. No support
for bioenhancement in this chapter.
The remaining chapters offer little	ntention that
bioenhanced the harbor ecosystem.	, „	, hapterV, B and
V, D include some results which seem to weaken the contention. The very
high mortality of anchovey larvae in diluted secondary treated sewage and
high mortality of mussels suspended in the harbor waters indicate intolerence
of the discharges, or problems of experimental design ot conduct, or
otherwise.
In summary, the report is not able to support the contention that the
harbor's biological environment was enhanced by the fish processing wastes
or the sewage discharges. An increase in the scope of the food web was
qJ	t>Cc\
not demonstrated while ana increase in the magnitude o£\the food web was
the cannery discharges and primary
j
treat ecr |^^^»^ischarges
have actually
suggested but not proven. There were some coincidental changes in

-------
populations which might presage the ecological results should the cannery
discharges be reinitiated. However, those changes (increases in white
croaker and polychaete worm abundance) arguably may not necessarily benefit
the entire marine ecosystem. Also, there is some indication that larval fishes
may be detrimentally influenced by the dischrges. It would seem that further
specifically designed, scientific investigation is warranted.
Due to the presence of two Federally listed endangered species in vicinity of
the outfalls in Los Angeles Harbor, the EPA may choose to consult the FWS,
under the provisions of Sec 7 of the Endangered Species Act, should a Federal
action be contemplated.
It is also worthy of mention that the area of^^MBqrroer cannery outfall
and present Terminal Island Treatment Plant outfall will be filled starting
in 1980. The Corps of Engineers and the Port of Los Angeles, as part of the
harbor deepening project,lintend to dispose of the dredged material by
creating 190 acres of new land out of the present relatively shallow
(less than 20 feet deep) area adjacent to Terminal Island. The Port also
proposes handreds of acres of fill in its Master Plan. The irony is that
should enhancement ever be convincingly proven, it could be for a harbor
environment that had already been destroyed by filling.
The delay in making these cursory comments is regreted. It is hoped that they
prove useful in EPA's review.

7
*. > >
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//
L.A.E. V. Cannery \sQfTITP Sewa.ce Owtfa.1l\
Qutfall^gyZ-	"	\
LA 19C
LA 10
\
• la 18a
Starkist #4 Outfall
LA 3A
H
1 MILE
LA 1
FIGURE 4.
VPvTER QUALITY STATIONS
Source; Los Angeles Haxfaor Dept.
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TABLE 1. 1976 - 19 77 SURFACE
Trans.	D.O.	BOD5
MONTH	(feet)	(mg/1) (mg/1)
19C 19D 19C 19D 19C 19D
JUN
1977
5
5
7.4
6.6;
7.3
1.4
MAY
1977
4
7
3.5
5.6
5.9
0.5
APR
1977
2.5
4
0.2
7.0
66.2
7.4
MAR
1977
3.5
3.5
4.0
5.1
, 12.6
4.7
FEB
1977
5.5
12
5.4
6.8
: 9.1
0.9
JAN
1977
7.5
5
4.7
2.6
7.7
11.7
DEC
1976
2
2
0.0
0.1
43.2
8.7
NOV
1976
4
7.5
1.5
0.5
3.2
2.3
OCT
1976
4
11
3.4
6.1
33
2.9
SEP
1976
9
3.5
6.3
3.0
! 5.9
20
AUG
1976
3
5
3.2
5.7
32
7.7
JUL
1976
2
6
5.3
7.3
¦ 	
	
AVERAGE
4.3
5.9
3.7
4.7
20.5
6.2
S = seme
C = considerable
F =» free
Si =* slight
* See Figure 4 for station locations.
Source: LAI ID Water Quality Survey.
QUALITY AT STATIONS 19C AND 19D*
Temp,	Oil 6	Floating
( C)	Odor	Color Grease Solids
19C
19D
19C
19D
19C
19D
19C
19D
19C
19D
18.5
18.2
F
F
YOG
OG
S
F
F
F
17.5
16.2
F
P
MG
MG
F
F
SI
F
19.0
18.0
FC
F
MG
LOG
F
F
C
S
14.0
13.4
F
F
MCI
MG
S
F
s
S
16.5
15.9
F
F
MOG
OG
F
F
c
c
17.2
17.0
F
S
G
MOG
F
F
c
c
16.8
17.4
F
F
LOG
LOG
S
SI
c
s
18.9
18.5
FCS
FC
LG
LG
F
F
SI
si
21.8
20.8
FC
. FC
LG
G
F
F
SI
F
19.1
19.6
FCS
FC
OG
LOG
F
F
F
SI
20.7
19.2
FCS
F
£OG
OG
F
F
F
F
21.3
20.4
FCS
F
G
G
F
F
F
F
18.4
17.8








M = milky	Y - yellow
G = green	FC = fish cannery
L = light	FCS = fish cannery and sewage
O = olive
-------
Los Angeles harbor Study
P. T. Erusaker
fT?'?*//??
Tne Files
2.
3.
On 4/5/79, at the Long Beach offices of the California
Department ot Fish and Game, an informal workshop was
convened to review the most recent (2/79) Draft L.A.
Harbor Study by the Harbor's environmental projects.
This study is a continuation of earlier work coiuF.issicnec!
by Terminal Island tuna canneries in order to aefine-
effects of cannery waste discharges. It has been
submitted by the canning industry as the industry's
contribution to the Section 74 Seafood i.aste Study,
being conducted by Effluent Guidelines Division.
The participants included:
^Jim Slawson, National marine Fishery Service
'AJack Fancher, U.S. Fish and Wildlife Service
•-HBoward bright, California hater Resources Control Ec-ard
Espinosa,"^California Department of Fisn and Ga-vc
Louis Schinazi, Los Angeles K'.'CCL
Cent- Dysir.oer , Section 74 Project Officer
-Terry Brubaker, Lnvironmental Protection Aoencv
-:L ,	¦¦
Althou^n no £orn.al conclusions were establisned regarcing
the stud^, a clear picture of its strengths and weaKnesses
developed over the course of the discussion. The attached
review criteria were used as a discussion guide.
Comments
A. Methodology:
The submittal is not oriented to fish waste Dis-
charges, but tov.aras defining the effects o£ 11^1
secondary effluent, it therefore cannot be reviewe:
witnout consiceration of earlier studies concucteu
prior to cessation of cannery discharges; the
reviews may then concentrate on the effects of
the absence of fish waste. The lack of suitable
Reproduced Irom
best available copy
C'.O
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-2- *
control areas could have been partially overcome by
comparison with data collected in Long Beach Harbor
by - consultants to Southern California EdiBon Company.
Several significant variables were excluded: other
nutrient sources such as runofr, weather patterns,
future harbor developments and long-terra fish
population cycles. Physical water quality data
were not included.
B.	Execution;
Documentation of field methods is lacking, but
most reviewers expressed confidence in the quality
of data. More importantly, field conditions were
not documented, including salinity, time, tide,
moon phase, temperature of air and water, cloud
cover, etc. Reviewers noted that general treli^*
of data would not be compromised by lack of such
documentation, but this information is critical for
understanding individual data points.
C.	Findings:
1.	Birds; Paul Kelly, Department of Fish and Game
bird expert, noted that data indicate an
improvement in the quality of bird population.
Grebes and pelicans, both fish eaters, increased
in 1978, while scavengers such as gulls,
decreased. The departure of gulls may also
lessen stress on endangered species in the area.
2.	Zooplankton: Data collected are inconclusive;
no significant effects were found.
3.	Phytoplankton: Data indicate no change in
populations but reduced assimilation rates
since cessation of cannery discharges. This
phenomenon is interesting, but cannot be
interpreted without an understanding of other
significant variables; i.e., an ecosystem
moael is necessary.
4.	Benthos: Reviewers agreed that this was tne
strongest portion of the study. It was noted
however, that the aata are improperly used as
a basis for speculation regarding effects on
fish populations.

-------
-3-
5. Fish: Reviewers agreej tnat the fish popula-
tion studies are incor.clusive. Stephens, the
- htp subcontractor tor trawl surveys, states7
in the report that "It is impossible to str.to
that there has been a detrimental effect"
from cessation of cannery discharges. Tne
stuiy did not reference other data available
from the 3CE Long Beach study and from recent
UbFtkS studies which indicate improvements in
fish populations since 1378. Reviewers
speculated that the various studies uid not
reflect anything more than a reaistriDution
of populations within tne harDor. She
study's statements regarding the impact oi
Harbcr population changes on fisheries
resources were- disccuntec by tne reviewers,
as were conclusions based on sport fishing
surveys, since these are not actually popula-
tion surveys, but are very rough "catch
effort" studies. Reviewers commented on
several inaccuracies in tne report, for
example, the statement that white croaker is an
important tccc resource sold as "butter!ish."
In fact it is illegal to so label white
croaKer, which is widely regarded as a trash
f i&h.
D	liJfuilo IT 'y .
'ine concensus ct the reviewers was that the study ?c
presenteo was a flawed ,	£ r agmentarv-ilocument. of limited
UtJ-lity fcr the purpose oi determining effects or fish
.waste or: marine waters. E;ven when considered with its
predecessor documents, it failed to conclusively demon-
strate a clearly defined effect. Among the reviewers,
tne Deportment of Fish end Game representative was most
critical, while the L.A.	biologist played the role
oi apologist to a decree. The problem of defining
"enhancement" was discussed, but no agreement was reached.
Reviewers will exchange written comments informally.
cc. :
V
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L. A. Harbor Enhancement Study Review Criteria
Section 74 of the Clean Water Act requires EPA to conduct a study of the
ecological effects of seafood wastes discharged to the marine environ-
ment. The seafood industry has advanced the concept of "enhancement" of
receiving waters by seafood wastes. This issue is of particular concern
in the L. A. Harbor area since "enhancement" is included as an exception
provision in the California Bays and Estuaries Policy (BEP). We request
that you review the enclosed document from two perspectives: first, technical
aspects and specific conclusions of the study; and second, evaluation of the
enhancement concept generally as it relates to L. A. Harbor and to water
quality goals and water uses. The following outline may be helpful in
your analysis.
Technical Review Criteria
1. Methology
A.	Adequate conceptualization in terms of logic, utility, scale
B.	Adequate study plan - accountability for all significant
variables
C.	Appropriate location of site{s)
D.	Clearly defined objectives
E.	Clear statement of qualifying factors
2. Execution
A.	Use of proper sampling and analysis techniques
B.	Sampling conducted under representative field conditions
C.	Completion of all required study elements
D.	Adequate justification for all changes to study plan
3. Data Reduction and Analysis
A.	Clearly stated findings
B.	Adequate statistical analysis
CT"
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C. The study as it applies to L. A.: how does it fit with future
harbor development plans; how does it relate to previous
research, what policy decisions does it indicate?
General Review Criteria
1.	Is the concept of artificial management of coastal environments
(i. e. "enhancing" various species populations) an environ-
mentally sound concept? Is an artificially managed marine
environment more desirable than a natural, pristine environment?
2.	If you feel that "enhancement" is a feasible concept, does the
subject study adequately define criteria with which to measure it?
3.	Other than savings in the form of decreased waste treatment
costs, are there any conceivable economic benefits to be realized
through implementing an ecosystem management program (i. e.
could harvestable species be enhanced thus increasing fisheries
yields)?
4.	Are you aware of any other work which conflicts with the subject
study with regard to the enhancement concept generally or with
regard to the study findings concerning trends in the Harbor's
condition over the past few years?
r-.3
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i.ATE OF CALIFORNIA—RESOURCES AGENCY
EDMUND G. BROWN JR., Governor
CALIFORNIA REGIONAL WATER QUALITY CONTROL BOARD-
.OS ANGELES REGION
07 SOUTH BROADWAY, SUITE 4027
OS ANGEIES, CALIFORNIA 90012
(213) 620-4460
October 1, 1979
TO: Mailing List
SUBJECT: Interagency/Fish Cannery Workshop on Issues related to the Discharge
of Municipal Effluent and Fish Cannery Wastes to Outer Los Angeles
Harbor
We plan to conduct a workshop on October 11, 1979, to examine the issues related to
the discharge of municipal effluent and/or fish cannery wastes to Outer Los Angeles
Harbor. The failure of the Terminal Island Treatment Plant to consistently and com-
pletely treat all cannery process wastes has prompted the need for a reexamination
of cannery/TITP/discharge relationships.
The workshop will be held in the Port of Los Angeles Meeting Room at the American
President Lines Terminal, located on Swinford Street (east of Harbor Blvd.) in San
Pedro (see attached map). The discussions will begin at 9:00 a.m. We cordially
invite your (or appropriate staff) attendance at the workshop.
All liquid process wastes from the fish canneries on Terminal Island are now secondarily
treated at the City of Los Angeles Terminal Island Treatment Plant (TITP), and treated
effluent is discharged by the City to Outer Los Angeles Harbor under NPDES Permit No.
CA0U53856. The canneries are currently allowed to discharge non-process wastewaters
directly to Outer Los Angeles Harbor and Outer Fish Harbor under non-NPDES waste dis-
charge requirements since there are no pollutants in them.
In recent raonths, particularly during the anchovy packing season, TITP experienced
difficulties in adequately treating all cannery process wastes, and was in repeated
non-compliance with its waste discharge requirements. This condition resulted In the
issuance of a Cease and Desist Order (79-133) to the City by the Regional Board on
July 23, 1979. In compliance with that Order, the City has prepared a report to the
Board specifying the cause(s) of non-compliance, and detailing certain proposed alter-
native corrective and preventive measures to bring the plant into 100% compliance with
its NPDES requirements (including the timing for their accomplishment). A draft copy
of the City's report (attachment 1) is enclosed.
The City maintains that compliance problems arose from the high BOD content of waste-
waters received from the canneries, despite dissolved air flotation (DAF) pretreatment
by these facilities, causing the design capacity of the plant to be exceeded. In addi-
tion, fluctuation in waste strengths and flows from the canneries caused significant
upsets in the biological treatment process. It is likely that violations of TITP's
requirements will continue to occur in the future at times when the plant's treatment
Gentlemen:
Background
-------
Page 2
Interagency/Fish Cannery Workshop
capacity is exceeded unless 1) TITP's treatment capacity is increased, 2) canneries
upgrade pretreatment and deliver a reduced (and constant) BOD waste load to TITP, jor
3) the canners are permitted to discharge the excess waste load directly to the Outer
Harbor on an interim basis. Alternatives (1) and (2) are costly and will require as
long as 56 months for implementation. Alternative (3) is fraught with technical and
administrative complications, as discussed below, but must be addressed as soon as pos'
sible to determine its feasibility as a mitigative measure.
Direct discharge of process cannery wastewaters to the harbor was prohibited by the
Regional Board in October 1977 because the canners failed to demonstrate adequately
that harbor water quality was enhanced by the discharge. Under the State's Bays and
Estuaries Policy discharges of municipal wastewater and industrial process waters to
enclosed bays and estuaries are prohibited unless the discharge would enhance the
quality of receiving waters above that which would occur in the absence of the dis-
charge .
One of the prime objectives of this workshop will be to obtain expert responses as
to:
(1)	the feasibility of considering any resumption of direct cannery discharge
within the framework of the existing legal constraints of the Bays and
Estuaries Policy and other State water quality standards.
(2)	the formulation of recommendations to the Regional Board (for later consider-
ation by the State Board) regarding interpretation of the existing Policy
in light of special conditions in Los Angeles Harbor.
(3)	given that the legal constraints may be resolved, under what conditions
could an interim discharge be allowed?
We look forward to your participation at the workshop.
I am enclosing a simple agenda and a copy of the City's report to the Board.
Very truly yours,
1
Executive Officer

cc: See attached mailing list
Enclosures
-------
Interagency/Fish Cannery Workshop
\
cc: Environmental Protection Agency, Region IX,
^Attn: Mr. Clyde Eller, Chief, Enforcement Division
[Environmental Frotection Agency, Washington, D.C.
Attn: Mr.\yJalvin Dysinger, EPA Seafood Study Project Officer
State Water Resources Control Board, Legal Division
Attn: Mr. William Attwater
Attn: Mr. Harry Schueller
Attn: Mr. Craig Wilson
Department of Fish and Game, Marine Resources Region
Attn: Mr. Rolf Mall
Attn: Mr. Larry Espinoza
U.S. Department of the Interior, U.S. Fish and Wildlife Service
Attn: Mr. John Fancher
National Oceanic and Atmospheric Administration,
National Marine Fisheries Service Attn: James Slawson
South Coast Regional Coastal Commission
Attn: Mr. Melvin Carpenter
City of Los Angeles, Department of Public Works
Attn: Mr. Jack Betz
City of Los Angeles, Department of Public Works
Attn: llr. Robert S. Horii
City of Los Angeles, Harbor Department
Attn: Ed Gorman, Chief Harbor Engineer
Port of Los Angeles, Attn: Calvin Hurst,
Chief Marine Enviorninentalist
Star Kist Foods, Inc., Attn: David Ballands
Attn: Anthony Nizetich
Pan Pacific Fisheries, Inc., Attn: Alan Pasarow
Attn: Joe McGrath
Dr. Dorothy Soule, University of Southern California
Environmental Protection Agency, National Enforcement
Investigations Center, Attn: Barrett E. Benson
-------
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Reproduced from
best available copy
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-------
AGENDA
Canners' Discharge Seminar
October 11, 1979
Introduction and Background
Problem
Alternative Solutions
Institutional Issues
Continuation of Study Program
C "3
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REPORT TO CALIFORNIA REGIONAL WATER QUALITY CONTROL BOARD,
LOS ANGELES REGION CONCERNING ORDER NO. 79-133
ON THE TERMINAL ISLAND TREATMENT PLANT
City of Los Angeles
Department of Public ".Vorks
Bureau of Sanitation
Bureau of Engineering
September 1979
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REPORT TO CALIFORNIA REGIONAL WATER QUALITY CONTROL BOARD, LOS ANGELES
REGION CONCERNING ORDER NO. 79-133 ON THE TERMINAL ISLAND TREATMENT PLANT
INTRODUCTION
Order number 79-133 of the California Regional Water Quality Control Board,
Los Angeles Region (RWQCB) was adopted on July 23, 1979 after a hearing
before that Board in which the reasons for the non-compliance of the treated
"effluents from the Terminal Island Treatment Plant with certain provisions
of its National Pollutant Discharge Elimination System (NPDES) Permit were
examined. Order number 79-133 directs that the City must meet the provi-
sions of the NPDES Permit and provides that the City present a written
report within a 60-day period (September 24, 1979) detailing to the RWQCB
the measures taken or to be taken, including an implementation schedule,
to assure that full compliance with the NPDES Permit limitations will be
achieved at the earliest possible date; and specific measures to mitigate
the effects of the discharge prior to achieving full compliance. Non-
compliance with the order would subject the City to injunction and civil
monetary fines.
At the time of the July 23, 1979 cease and desist hearing, the City pre-
sented information to the RWQCB which in its opinion showed the compliance
problems to arise from wastewaters discharged by four fish canning install-
ations connected to the sewage system tributary to the Terminal Island
Treatment Plant. It indicated that the wastes so received exceeded
design capacity and were in addition discharged in a manner such that
extreme cycling of waste strengths was present. These factors were such
that the ability of the biological treatment processes to adjust was
exceeded and violations had therefore occurred. It was, however, noted
that in general, treatment had been effective and that receiving water
standards had been met.
STAFF PARTICIPATION
Noting that the problem was a complex one, the City stated its intention
of working closely with representatives of the canneries in seeking a
solution. It also requested that the RWQCB authorize personnel from its
staff to work with the City and the canneries to insure that appropriate
alternatives would be reviewed within the time required. The RWQCB agreed
that such participation by its staff would be of value and it was so
authorized.
BASIC DATA
Review of the plant performance has shown that the critical factor causing
non-compliance with permit requirements has been the high organic loads
received. Although standards for parameters other than BOD^ have also been
exceeded, the actual organic load can be best characterized in terms of
BODr. The available data covering the basis of design for the Terminal
Island Plant, the amount of organic loading planned for each type cf dis-
charge and the amount and nature cf the fish cannery loading in terms of
BODj. is accordingly summarized below:
1. Design Capacity:
a.	Present Plant	53,600 lbs/day
b.	When Unit IIA (Solids Handling)	80,000 lbs/day
completed. _
C'O
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2.	Current Domestic Input	20,000 lbs/day
3.	Planned Cannery Input From Consultants Harbor Study: 19,600 lbs/day.
From these it would appear that the capacity should be adequate since design
capacity was 53,600 lbs/day and the projected current loadings was about
40,000 lbs/day. Figure 1 and Table I attached, show the problem which has
developed inasmuch as daily loadings from all sources of organic loadings
have often exceeded 100,000 lbs/day. It will be noted that with organic
loadings greater than 100,000 lbs/day of BODj. compliance was achieved only
about 26% of the time whereas at the 53,600 lbs/day design loading com-
pliance was achieved more than 90% of the time.
Completion of the unit IIA (Solids Handling System) contract in December
1981 should increase the average capability of the plant to handle organic
loads from 53,600 to 80,000 lbs/day or by an additional 26,4 00 lbs/day.
Assuming all of this could be dedicated to the cannery wastes for a period
until other loadings increased, the plant capacity available for this use
would be approximately as follows:
80,000 lbs/day	Ultimate Plant Capacity
Less 20,000 lbs/day	Domestic Wastewaters
Less 3,000 lbs/day	Other Industrial Wastewaters
57,000 lbs/day	Canneries
Since daily loadings on the treatment plant attributable to the canneries
have frequently exceeded the ultimate total design loading figure of
80,000 lbs/day, the 57,000 lbs/day which could be made available to the
canneries under the conditions expected to prevail when the present con-
struction work is completed would still be insufficient. There could
therefore be no assurance of complete and continued compliance with the
NPDES regulations if all cannery wastes continued to be received by the
treatment plant. In addition, since fish catch is controlled both by fish
abundance and by fishing days set by the State Department of Fish and Game,
it is quite probable that organic loadings either in excess of those
received to date or persisting for a longer period would occur. Another
factor needing evaluation is whether the increase arising from completion
of present facilities can be permanently committed to the fish canneries.
The alternatives will consider this.
ALTERNATIVES
Given the facts outlined above, the following are the alternative proce-
dures which could be followed to achieve the required 100% compliance with
the NPDES Permit regulations:
1. Modify the Treatment Plant as a grant fundable project to allow the
heavier loadings to be properly treated. This might include the
following:
i
a.	Retrofit the aeration system to allow a fine bubble aeration or
high purity oxygenation system to be installed.
b.	Provide flow equalization tanks to provide a more uniform loading
to the plant.
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120
110
100
90
BO
70
60
50
t
40
30
2.0
10
0
FIGURE I


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sr;:
-v-

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535=

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-+-


 SUBTACTING VIOLATIONS ARISING
FROM LOW AIR OR MAJOR PEAK
LOADS (UPSETS) PREVIOUS DAY
-T
iM
:nr-
:zi:
—rr
PPI
ebjei:::;
*; 11L

t * "hi"? H*-h
# i
' ii JJ
II
-I ; 1 I
P - t-
7];-j li
Hi t1J H t
* " ' ? H t r
rr; > • 11 ii
. ,.i , j i , , ¦
)	20 30 40 50 60 70 80	90
PERCENT COMPLIANCE (EFFlUENT < !0,000 LBS B0D5 /DAY)
-3-
I00
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TABLE 1
TERMINAL ISLAND TREATMENT PLANT.
1978-79 DATA ANALYSIS
BOD Mass Emission Violations
Raw Influent No. of
lbs/day Days
No. of Days Effluent
Exceeded 10,000 lbs/day
% Compliance
100/000 26 days
19
26.9
90,000-100,000 9 days.
4
55.6
80,000-90,000 14 days
7
50.0
70,000-80,000 17 days
6
64.7
60,000-70,000 32 days
6
81.2
50,000-60,000 51 days
5
90.2
40,000-50,000 55 days
5*
90.9
30,000-40,000 67 days
3**
95.5
20,000-30,000 50 days
2**
96.0
10,000-20,000 44 days
0
100.0
365 days
59 days

*Two of these five violations were due to insufficient air being provided
to aeration tanks because of clogged diffusers.
The other three were due to delayed reactions from much higher peak
loads on previous days.
**Violations due to delayed reactions from much higher peak loads on
previous days.
r 3
-4¦
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C. Provide additional secondary sedimentation and sludge handling
facilities as necessary.
It is believed that it may be possible to justify this approach inasmuch
as neither the City, the industry, the State nor EPA had valid data to
show what a fish cannery loading would be at the time of design, given
the many types of fish handled. Also, EPA pretreatment standards had not
been set and the strength of the effluents were not known, given the
variety of fish and the variability of pre-treatment effluents from each
species packed. The projected plant change could then be a simple adjust-
ment to fact rather than a growth inducing step.
It is proposed that fine bubble jet aeration system utilizing air and/or
high purity oxygen, such as that now under test, be evaluated with the
appropriate sedimentation tanks ana adequate aerated equalization tanks.
Capital costs are estimated to be of the order of $6,000,000.
The time required to accomplish this is estimated as follows:
Step
1:
Project Report and Approvals
6
months
Step
2:
Design and Approvals
12
months
Step
3:
Contract Award, Construction and other
State and Federal Approvals
36
months


Total Time
54
months
Thus if Step 1 could be started by January 1, 1980, operation would
be by June 30, 1984. All fish cannery and other industrial and
domestic loadings could be handled by this alternative. Alternative
2 could be used in the interim for all loadings greater than the plant
facilities could treat.
The Terminal Island Treatment Plant could accept organic loading from
the canneries on each day up to approximately 30,000 lbs. until 1982
and then up to 57,000 lbs. from that point on with the canneries dis-
charging amounts in excess of allowed loadings to the harbor through
a new cannery outfall which would provide enough diffusion to prevent
a marked dissolved oxygen sag in the discharge area. (Increased
loadings from domestic sources and other industries could change
this entitlement.)
a.	The studies of enhancement, to the fisheries in the harbor as
a result of such waste discharges to date has indicated that
probability that enhancement is present. Comments by California
Fish and Game and other agencies had indicated that certain
further areas of study as to enhancement would be valuable and
should be completed.
b.	Impact on Harbor waters would be limited by using the treatment
plant to capacity and directly discharging only excess loadings
through a new cannery outfall-aiffuser system. The excess amount
would, however, allow the desirable enhancement study to proceed.
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C. If enhancement could not be shown then the canneries would be
obliged to help implement either Alternatives 1 or 3 described
herein.
The time necessary to implement Alternative 2 is estimated as
follows:
1. Obtain necessary environmental approvals
6 months
2. Design and construct outfall and diffuser
8 months
3. Total enhancement study time
36 months
Total Time
50 months
Thus, if the process could be started by January 1, 1980, the new
outfall and diffuser would be in operation by March 1, 1981 and the
full study completed by March 1, 1984. If enchancement was demon-
strated, either this alternative or Alternative 4 would then continue
Capital cost for the construction of the outfall and diffuser and
the necessary facilities is estimated to be $1 million.
The third alternative would be for the City to limit the canneries to
approximately the 30,000 lbs/day of BOD^, which would bring the plant
to current design loading, until 1982 and to approximately 57,000
pounds per day after that date when Unit IIA (Solids Handling Unit)
is completed. This would place the full responsibility on the canneries
to meet these loadings by steps such as providing further pretreatment,
flow control and other internal procedures. (It also might be neces-
sary to reduce the cannery allotment if other domestic and industrial
loadings were received.)
In attachment A to this report, the canneries have given their response
to this alternative giving background data on plant capacities and
corresponding waste loadings and indicating the effects that their
resulting operational reductions would have upon employment levels
in both the canneries and the fishing fleet.
The fourth alternative would be for the canneries to discharge their
total waste flow with non-process waters to a new cannery outfall-
diffuser system designed to prevent an oxygen sag and excessive
bottom deposits.
a.	This could aid the enhancement study.
b.	The ability of the harbor to handle the full load of properly
diffused organic wastes could be tested.
c.	If enhancement is shown the procedure could be followed with no
change needed.
The time necessary to fully implement Alternative 4 is estimated
as follows:
1.	Obtain necessary environmental approvals	6 months
2.	Design and construct outfall and diffuser	8 months
3.	Enhancement study time	36 months
C 5
-L, -
Total Time
50 months
a
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Thus if the process could be started by January 1, 1980, the new
outfall and diffuser would be in operation by March 1, 1981, and
the full enhancement study completed by March 1, 1984. If enhance-
ment was demonstrated, the alternative could then simply continue.
The canneries have commented on this alternative in Attachment A
giving a brief background of some related actions.
Summary of Alternatives
Figure 2 summarizes the four alternatives showing their relation-
ships with the various decisions involved. Also shown are two
interim discharge options and the planned Interagency/Canners
Workshop.
"RECOMMENDATION:
k decision as to the ultimate choice of alternatives cannot be made until:
1.	The Regional Water Quality Control Board, the State Water Resources
Control Board and U. S. EPA Region IX can determine whether a con-
tinuation of the present study on enhancement is in order.
2.	If such a continuation is considered pertinent, whether enhancement
is or is not shown at the time of its completion.
From the factors cited above, two avenues of approach are possible. Thus,
if a continuation of the study is not judged to be productive, alternatives
1 or 3 must be undertaken. However, if continuation of the study on enhance-
ment is judged proper, any of alternative 1, 2, 3, or 4 might eventually be
adopted.
Regardless of which of these decisions is made, an interim period will exist
before the necessary permits and/or construction can be accomplished. Opera-
tion in the interim must therefore also be considered. With these factors
in mind, the following procedure is recommended to the RWQCB:
1. The Executive Officer of your Board is organizing a workshop to include
representatives of your Board, the State Water Resources Control Board,
U. S. EPA Region IX, Federal Fish and Wildlife Service, California Fish
and Game, the canneries, the City and other interested parties to
explore the question of the value of further research upon enhancement.
This will be directed at enabling a decision on this important point to
be reached at an early date.
_2. An interim period will exist before such a decision can be reached; and,
if the decision is that further enhancement study is desirable, before
necessary permits, design and construction of a new cannery outfall-
diffuser can be accomplished. During this period, the following is
recommended:
a.	Utilize the Terminal Island Treatment Plant to treat the greatest
cannery organic load possible while staying within NPDES Permit
limitations.
b.	Use the present cannery outfalls to the outer harbor area for the
balance of the non-process and process wastewater from their
facilities.
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If an extended enhancement study is recommended, and after a new
outfall-diffuser system is completed, put all cannery wastewater to
the harbor through the new outfall-aiffuser system during the period
of the study (alternative 4). This alternative would be subject to
further modification during the study period should it prove more
desirable to reduce the cannery discharge to the harbor and direct
some of the wastewater to the Terminal Island Treatment Plant.
If an extended enhancement study is not recommended or if after the
study enhancement is not shown, enlarge plant capacity to handle
these wastes (alternative 1).
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WITH HPDES WASTE D1SCUA. it Kti^uintntu13
INTERIM DlSCHARGt
(two options) '
	I	
1. a) TITP treats up to
30,000 lbs/day BOD
b) Canners discharge excess
to harbor (DAF-treated)
2. a) TITP continues to treat all
pastes and discharge with
periodic violations
b) Canners reduce BOD loading
to TI-TP and equalize flows
to minimize TITP violations
(short-term solution)
1	
EBiancement may occur, but
not yet sufficiently demonstrated
f*use 3-year Study Plan under
lanaged discharge conditions
^>lement Study - January 19o0
Allow Interim Discharge
Wteragency Review of Data
Annual Reviews
Final Review
Decision
(ALTERNATIVE k
All Cannery wastes
to harbor
If enhancement dees occur
TITP discharges excess
via bypass
Canners discharge excess
thru new outfall
Enhancement does not occur;
Further studies not warranted
Long-term Solutions
Allow Interim Discharge
pending completion.
Managed Discharge only
30-57,000 lbs/day to TITP
Excess to harbor
ALTERNATIVE 2
• ALTERNATIVE 1.
TITP upgrades to >80,000 lbs/day
peak capacity. Continue to handle
all wastes from canners
ALTERNATIVE 3.
Canners upgrade 6 improve
pretreatment
Limit production to meet
City effluent limits
IMTERAGEIJCY/CAHMERS WORKSHOP
To review evidence for enhancement
within context of recent State
Board interpretation of Bays & Estuaries
Pol icy
Workshop-October l|, 1979
Decision-December 1, 1979
Reproduced from
best available copy,
r "3
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ATTACHMENT A
Comments by the Fish Canneries
Relative to Alternative Nos. 3 & 4
1. In consideration of Alternative No. 3, which fixes limits for the
cannery discharges to the treatment plant, it is valuable to compare
the effluent loadings from each plant historically since the intro-
duction of DAF with the loadings considered typical for the industry
related to the production capacity for each plant.
-
Current
Capacity
BOD Loadings 1
Calendar Year 1978
(lbs./Day)
Standard Industrial
Performance for
Average 4
(lbs./Day)
Plant
(Tons/Day)
Avg.
Peak
Current Capacity
Pan Pacific
#1 & #2
375 Tuna
350 Mackerel
40 Pet food
300 Anchovy
3,200
10,000
6,750
6,300
720
2,340
16,110
Pan Pacific
120 Tuna
300 Anchovy
1,164
8,0002
2,160
2,340
4,500
SKF #1
1,200 Anchovy
480 Mackerel
135 Pet food
5,460
27,4002'3
9,360
8,640
2,430
20,430
SKF #4
675 Tuna
38 Pet food
7,900
20,000
12,000
684
12,684
NOTE:
Due to metering errors, flow may have been underestimated; due
to differences in analysis, BOD concentrations may have been
overestimated.
2	Anchovy production was low during 1978.
3	During peak production days in 1979, peak loadings were
approximately double those shown.
4	BPT performance - used by RWQCB in 1977 Cannery NPDES Permit.
Tuna (CFR 408.142)
Mackerel (No Ref.)
Pet Food (No Ref.)
Anchovy (CFR 408.152)
Solubles Plant
Other
Max.
Avo.
231b7l"0~00 lb. 9.01b7l000 lb.
If
N
7.01b/1000 lb. 3.91b/1000 lb.
3.51b/1000 lb. 2.81b/1000 lb.
CO
A-l
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It is evident that the peak loadings from Star-Kist during the high
anchovy production periods in 1979 were atypical and the reasons for
these peak loadings will be determined and dealt with. However, it
is evident that if the peak daily loading of 23 lbs/1,000 lbs* allowed
by the July 1, 1977 EPA guidelines were applied to all the four plants
products but anchovy, and using a value of 7 lbs/1,000 lbs* for
anchovy, then a total cannery capacity of about 125,000 lbs per day
would be required for current capacity levels; i.e. an additional
95,000 lbs per day over 30,000 lbs per day which may be available
for the interim period until the municipal treatment plant is com-
pleted. The canners accept that they can improve the operation of
their plant and their DAP systems so as to reduce their effluent
loadings on a pounds per ton of production basis compared with those
which have occurred over the last 18 months. However, the general
concensus is that this degree of improvement will be considerably
short of providing the solution to the basic equation of demand vs.
treatment plant capacity. Therefore, the canner's response to the
above situation can best be forecast as follows:
a.	Facilities which will be required for the canners to reduce
their loadings at each plant to the level required would be
equivalent to secondary treatment and similar to those required
to treat the waste by expanding the municipal treatment plant.
b.	The time required to provide these facilities would be roughly
equivalent to that required by the Municipal Treatment Plant
to extend its facilities.
c.	The Municipal Treatment Plant receives considerable dilution
water with which to handle the high cannery loads; the canneries,
of course, have no dilution water and, therefore, the technical
difficulties in treating the waste would be greater if it were
treated at a cannery secondary treatment plant and cost of the
facilities is likely to be greater.
d.	The canneries have no space in which to install secondary
treatment facilities.
e.	After considering the probable impact on profit contributions
and employment, the canneries would probably choose to operate
that portion of their current production which could be
accommodated within the allotted BOD loadings and would be
forced to cease to operate the remaining portions of their
plants (all types of fishing are considerably interdependent,-
and it is unlikely that the fishing fleet could continue to
exist in its present form). Apart from the loss of income to
the canners, the Port of Los Angeles, and the impact on local
ancillary industry, the resulting loss of employment would be
approximately as follows:
* These are the loading rates used in the cannery 1977 NPDES permits.
It should be noted that the loading factor for tuna was withdrawn
by the EPA in the August 6, 1979 Federal Register based on new infor-
4-hi«5 1977 BPT limit was not consistantly achievable. ^
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Personnel employed in the fishing fleet - approximately
500 (plus ancillary business, e.g. ship repair, marine
supply)
People employed in the canneries - 1,000 to 2,000 (plus
ancillary business)
(NOTE: For statistical purposes ancillary business employment
is calculated by a factor of 5 or more.)
In consideration of Alternative No. 4, which would allow discharge of
all cannery wastes directly to the harbor, flows and loadings from
the canneries have decreased over the recent years due to improved
handling and conservation measures within the canneries. The install-
ation of the DAF units has further reduced loadings being discharged
from the canneries. The various work being carried out by USC and
others over the past 10 years and reported in the recent summary report
prepared by the Allan Hancock Foundation suggests that a considerable
body of evidence exists to support the concept that these wastes,
which are principally of ocean origin and are biodegradable, bioenhance
the receiving waters. Further work which has been done to reduce
cannery effluent loadings since much of the research work was carried
out suggests that there is good reason to reconsider the case for
bioenhancement. It is suggested that this should be progressed in
the following way:
a.	A series of meetings should be convened between the industry,
the City of Los Angeles, the Regional Water Quality Control
Board and the Environmental Protection Agency to determine the
experimental design for data to be collected during the pro-
posed period of monitored discharges from the canners to the
Outer Fish Harbor.
b.	Design should be progressed for a new cannery discharge system
into the Outer Harbor.
fi-3
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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
REGION IX
215 Fremont Street
San Francisco, Ca. 94105
In Reply E-5-2
Refer To: 863.4A
Raymond M. Hertel
Executive Officer	0 2 OCT
California Regional Water Quality
Control Board
Los Angeles Region
107 S. Broadway
Los Angeles, CA 90012
Dear Mr. Hertel:
This is to provide you with comments on proposed agenda
for the Los Angeles Harbor workshop scheduled for October
11, 1979, as discussed in your letter of September 11 and
at our meeting of September 25.
Our primary concern at this time is to insure that all
participants fully understand the statutory and regulatory
framework for the discussions. The Bays and Estuaries
Policy specifically prohibits new discharges to the harbor.
Any application for a permit for such a discharge must,
therefore, include a positive demonstration to satisfy the
"enhancement" provisions of the policy. The policy does
not provide for a prospective determination of enhancement.
Furthermore, our review of the most recent available update
of the Harbors Environmental Projects study does not indicate
that such a positive demonstration of enhancement can be made
at this time. With this in mind# we believe that item IV on
the agenda (interim harbor discharges) may represent an area
of academic interest, rather than a viable regulatory option.
We believe it would be most useful to focus on pragmatic
issues, including:
1} The nature of TITP operating problems;
2)	Mitigating measures taken and planned at TITP
and the canneries;
3)	Schedules and costs of deepwater outfalls;
4. Regulatory options.


/
These would most appropriately fall under item I of the agenda.
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(2)
While we are willing to discuss the subjects covered
in Item II, it may be that a one-day workshop would not
allow sufficient time for a meaningful examination of
each of the technical points listed.
If you wish further discussion of the agenda items, please
contact Terry Brubaker at {415)556-7841.
Sincerely yours,
L. x_y uu lj • i-< j. -lca.
Director
Enforcement Division
cc: SWRCB
hoi UUbf* ¦ ItoM'SSd)
10
C 3
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TATE OF CALIFORNIA—THE RESOURCES AGENCY
EDMUND G. BROWN JR , Go*f
ATE WATER RESOURCES CONTROl BOARD
. O. BOX 100 • SACRAMENTO 95B01
(916! 445-7971
HViSlON OF WATER QUALITY
AUG 7 1973
Mr. Calvin Dysinger
Effluent Guidelines Division
In Reply Refer
to: 526: 1W
Environmental Protection Agency
401 M Street, SW
Washington, D.C. 20460
I have alreadj' forwarded our Division's comments on Marine Studies of San Pedro
Bay, Part 16, and on the two environmental impact reports that incorporated that
document. Those comments reflected our Division's point of view, and concentrated
on issues relevant to the Clean Water Grants Program.
My own professional opinion of the Section 74 program follows, in the format
requested in your letter.
Methodology
The document reports on research undertaken to assess possible enhancement by
secondary effluent from Terminal Island Treatment Plant, not the discharge of
cannery wastes directly to Los Angeles Harbor. The conclusions of the study may
bear on the question of direct discharge of cannery wastes. However, since the
City's treatment plant discharges close to the former cannery outfalls, it is
hard to distinguish the effects of changes in the City's effluent from cessation
of the cannery discharge.
I believe, from my observation of the collection of samples, that the ecological
sampling was done carefully and with generally appropriate techniques. Our
comment letter notes some disagreement on techniques.
Data Reduction and Analysis • • ¦
I did not feel that the report's findings follow clearly from the data. Specific
problems are noted in our comment letter. One problem was that some of the most
critical data, such as measurements of nutrients, were presented only indirectly
and incompletely. The reader is asked to take on faith several statements that
photosynthesis is not nutrient-limited. These statements seem to conflict with
the assertions that secondary treatment of wastewater has deprived the harbor of
nutrients needed for photosynthesis.
The study does fit in with the next planned phase of harbor filling. It points
out that further filling beyond the phase scheduled for 1980 will so damage har-
bor circulation that water quality will probably be degraded. That point is
relevant to the question of discharge of cannery wastes.
Execution
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-2-
General Review Criteria
My personal opinion (as distinct from that of SWRCB) is that the concept of
"'enhancing' various species populations" in the ocean is environmentally sound.
We already do that in an unplanned way by fishing out predators, such as tuna.
However, any attempt at enhancement should add only materials that are known to
be non-toxic and easily biodegradable. Some of the Los Angeles fish cannery wastes
showed toxicity on standard bioassays and the source of the toxicity was apparantly
never found. I can conceive of a situation in which cannery wastes, properly
diffused, could enhance Los Angeles Harbor.
I do not think an artificially managed marine environment is as desirable as "a
natural, pristine environment". That judgement follows both from my personal
values and from the fact that I have little confidence that we will manipulate
the marine environment wisely or well. However, Los Angeles Harbor is not "a
natural, pristine environment". There is evidence that the whole Southern
California Bight is profoundly changed from its natural state.
The subject study defines "bioenhancement", not enhancement, and its list of
criteria on page IB3 omits mention of primary productivity, a critical issue in
local waters. The concept of spatial averaging set forth elsewhere in the
document is highly controversial.
The study sets forth criteria with which to measure "bioenhancement", but they
are not stated such as to allow a "yes" or "no" answer from the data to be
gathered. Operationally definable criteria are necessary to a decision.
I agree with Soule and Oguri that cannery wastes could conceivably be discharged
to the harbor in such a manner as to enhance populations of fishes. I find it
distressing that in years of research no one has directly tested the idea that
anchovy schools feed on cannery wastes. I believe that such a test would be
simple and inexpensive if it were well designed, including proper handling of
the anchovies. I suspect that schools of anchovies actually fed on the cannery
wastes in 1976, but no really serious effort was made to test that commonly
held belief.
I believe you are already aware of the information on harbor fishes gathered by
the California Department of Fish and Game, National Marine Fisheries Service,
and Marine Biological Consultants. I feel that others on the committee are more
competent than I to comment on the data.
In summary, I believe that we still lack a clearcut study of the effects of
cannery wastes on Los Angeles Harbor. I personally think it possible that a
separate discharge of cannery wastes, if properly diffused, would benefit
anchovy populations without harming the harbor ecosystem. I believe that the
necessary research to evaluate that possibility remains to be done. The HEP
studies as a whole could help evaluate the possibilities, but are insufficient
to answer the questions posed by Section 74.
Howard 0. Wright, Ph. D.
Environmental Specialist
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BIMMON C. FAY. PhD,
TO Cal Dysinger
16 June 1979
In 1963 I proposed a way ftf poolii
data collected in the inshore area fo:
common reference. This suggestion re;
in SCCWRP but not in the pooling of th<
data. As a result, we still have obsei
-al data being collected and presented
individual format by non-standardized
technique for specific purposes by non-|
reviewed investigators. SCCWRP has nev<
approached this problem.
Would EPA be interested in our organi:
a conference for discussion of the subjet
and could EPA sponsor such a workshop?
This is important for CZK purposes, NMFS
management programs, EPA review of waivet
applications, and for the marine biologis
working in the area as well as CEIP studie
I can* provide an agenda, meeting plac
chairperson, and contribute to such a work
if it is of interest. We just did one on
coastal wetlands that worked out very wel
I hope that your find our comments on
the latest by Soule and Oguri to be of int
and of some use.
Best regards
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a
PACIFIC BIO-MARINE LABS, INC.
P.O. BOX 536
VENICE, CALIFORNIA 90291
June 14, 1979
TELEPHONE
OFFICE (213) 822.57!
Cal Dysinger
Environmental Protection Agency
401 "M" St. SW
Washington, D.C. 20460
Dear Mr. Dysinger:
Enclosed please find the comments of J. A. Vallee and my-
self on Ecological Changes in Outer Los Angeles - Long Beach
Harbors Following Intiation of Secondary Waste Treatment and
Cessarion of Fish Cannery Waste Effluent edited by D. F.
Soule and M. Oguri, April 1979.
These comments follow two approaches, direct comments on
the studies presented by Soule and Oguri and comments of an
informal nature based upon our own observations over the
same period (1971-1978) and taken in the harbor of Los Angeles
Long Beach as well as elsewhere along the coastline of
Southern California. This is followed by some recommendations
for your consideration with regard to the development of
criteria whereby to measure biological changes that may re-
late to changes in water quality along the coast of Southern
California.
We have enjoyed being able to volunteer our comments upon
these reports on cannery waste and the Los Angeles-Long
Beach Harbor complex. I note that this volunteer service
has taken many days of staff time and has resulted in a
determination that the volunteer effort cannot continue in
the form which it has taken in the past. However, this is
not the major problem; the problem is how to develop informa-
tion which your agency can rely upon and which will sustain
solid scientific review? Obviously, we should be grateful
for an opportunity to develop such information based upon
our extensive experience in this area and the development of
specific research programs designed to answer the questions
which beset the EPA about wastes discharged into the inshore
area.
We share the concern of Soule and Oguri in evaluating the
significance of the on-going change in water quality in the
harbor area and delight in what is happening as a result of
the abatement of toxic discharges here and elsewhere in the
Southern California Bight.

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June 14, 1979
Page two
Please advise us of how we may be able to continue to
cooperate with the EPA as your programs continue to im-
prove the quality of our inshore waters.
Yours sincerely,
Rimmon C. Fay
CC: James Rote, N.M.F.S.
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Two separate approaches to an understanding and manage-
ment of marine resources conflict in evaluating the report of
Dr. Soule and her colleagues. One assumes that the nature of
the quality of ocean waters is that of a highly oxidized,
highly mineralized, non-toxic medium with low concentrations
of bacteria, particulate and dissolved matter, and with
moderate to high transparency in the water column. This is
the quality of the medium in which most marine organisms
evolved and it is the quality of waste water adequately
treated before discharge to the ocean. Dr. Soule and her
colleagues argue for the discharge to the ocean of inade-
quately treated wastes (incompletely oxidized and mineralized)
with high concentrations of bacteria, suspended solids, plus
possible toxins, and for the development of an ecosystem
founded upon this unnatural and illegal basis.
Recognizing the objective of the Clean Water Act to
make the waters of the United States Fishable and Swimmable,
the studies in question are presented by Soule, et.al. from
the aspect of the fishability ignoring the considerations of
swimmability and the maintenance of a balanced endiginous
biota. One could not recommend swimming in the cannery
waste and the evidence for a balanced endiginous biota is
based upon an ecosystem of scavengers.
As discussed in our review of the preliminary report,
the authors establish their own definitions and assumptions
and then proceed to develop the types of evidence needed to
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give the trappings of a scientific approach to the proof
of their case. This strategy produces far more heat than
light as well as ignoring the basic legal issues involved
while providing some pap as legitimate well founded scientific
investigation. It becomes repugnant for the inexperienced
reviewer to deal with this quality of material and absolutely
revolting for anyone who would appreciate better work being
done in this area or anywhere else.
p.vii - "The Harbor was, in '73-74 the richest soft-
bottomed marine area in southern California"... on what
basis? comparisons? at what depths? how much of the bottom?
A reduction in the abundance of fishes is noted in the
period from '73 to '78 but similar observations were noted
elsewhere in Southern California and anchovies especially
declined in abundance everywhere during this period (see
Stevenson, p. 43, last quoted paragraph in this report).
p.vii - Fish are attracted to solid structure on the bottom
or at the surface and into areas of turbulence in the water
column. This cannot be considered enhancement per se as they
may be attracted to a fish trap just as well as this is the
basis for functioning of fish traps and gill or trammel nets.
p. ix - A 30 fold drop in the concentration of bacteria,
fungi, protozoa, etc. after implementation of secondary
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waste treatment would be regarded as an improvement in
water quality by any known criteria for evaulating the
suitability of water for the sustenance and maintenance
of fishes.
The statement that the bioassays demonstrated no evidence
of toxicity of the secondary effluent is predictable to
organisms "Typical of harbor wastes" (PIB4) were used? how-
ever, the techniques were questionable even so. Very
tolerant species lived under all conditions and the more
sensitive species died under all test conditions including
the control conditions. In addition, sub-lethal or chronic
effects were not examined.
ix - The trend from a bacteria/detritus based food web to
a phytoplankton based food web can be considered as an
improvement in water quality to support more natural balanced
endiginous biota.
xi	- The TITP is important to maintaining a population
of scavenger fish in the harbor but what does this do to the
function of the protected waters as a nursery and how does
this compare with unpolluted protected waters such as Mission
Bay?
xii	- Benthic resources might be expected to change in
specific abundance following an initiation of secondary
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treatment which is what this change was intended to
accomplish. The fact that a change in abundance of those
species typical of natural unpolluted conditions is going
to be a slow process is also predictable because bf the
more than 50 years of degradation of water quality in this
area few of the sensitive species are left to recolonize an
unpolluted area and no programs for active restocking are
being implemented.
xiv	- Increased concentrations of suspended solids are
reported to result in increased rates of growth of mussels
but it is not evident that this is equivalent to an increase
in a balanced endiginous biota.
xv	- The time frame considered, 1971 - 78, begins after more
than 50 years of artificial alteration of this area and
adverse impacts upon water quality from sewage, toxins, in-
dustrial wastes, cooling waters, and cannery wastes. At-
tempting to relate the biota of the harbor waters to the
activation of secondary treatment at the TIPT plant ignores
the massive other changes which have occurred in the inshore
waters as a result of pollution and physical modification of
the habitat.
xvi	- The toxicity of cannery wastes is discounted at the
expense of ignoring the unnatural effects of high concentra-
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c r*
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tions of particulate matter, lowered pH, reduced concentra-
tions of dissolved oxygen, and reduced transparency in the
water column.
IA9 - Definition is required for the term nutrients and
for the context of the use of the term. In the last para-
graph, natural or artificial upwelling is taken as equivalent
to artificial discharges and the equivalence of artificial
discharges to natural run-off from the land is implied. All
of these processes and the components involved require defini-
tion and qualitative as well as quantitative comparison.
IB4 - Even though "no 'control' harbors" are available, at
least some qualified comparisons should be made with other
areas of similar sediments, temperature, salinity, depth,
_ftc. that are not subject to waste discharges.
IC4 - Unofficial rainfall figures are used. Why not use
official records which are more complete and are accessible
to the public?
It is noted in reviewing this report, that the fish biota of
the bay drops as the studies continue but the impact of the
studies on the icthyofauna is not evaluated.
There is something very curious about the structure of the
icthyofuana of the bay as detailed by these studies. Very
few elasmobranchs are taken and while the absolute numbers
C 3
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of fish taken varies from year to year, as natural varia-
tion occurs, the relative abundances are essentially
constant save for anchovy but many of the formerly abundant
fishes on this coast are completely absent. These include
barracuda, pacific mackerel , pacific sardine, yellowtail,
bonito, and white sea bass. The absence of these species
is not even discussed in relation to those species which are
observed.
II A 2-3 - These studies concentrate on fin fish which
feed on garbage .... is this enhancement? Fish are also
attracted to thermal outfalls and into the intake structures
where they are killed as a result	is this enhance-
ment?
IIA 5 - The data base for the estimate of the abundance
of anchovies is inconsistent. It is estimated that they
are reduced 100 fold in 1978 versus the period of '72-73
based upon trawl surveys. Offshore a reduction of 4 fold
is estimated based on accoustical surveys in the region of
greatest natural historic abundance of these fishes.
Acoustical surveys are ofetn inaccurate; trawl surveys have
not been conducted off-shore.
IIA 17 - If the icthyofauna of the harbor is defined in
terms of those species utilizing the wastes from the cannery
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discharges, then cessation of the discharge of these
wastes may alter their local abundance at the site discharge.
This does not establish that their overall abundance declines
or that the cessation of the discharge will not result in
increased diversity of fishes or improvements in water
quality will improve the nursery function of the harbor.
IIA 21-22 - The fact that birds feed on the cannery wastes
is considered as enhancement and rats have been observed to
feed on garbage; is this enhancement?
IIB - The wide fluctuation in relative abundances of birds
in southern Californi has been observed for years. In fact,
it is only recently, 1977 or so, that good quantitative
information has been developed for many species of birds
which may be seen on this coast. Depressions in the abun-
dance of cormorants, pelicans, bald eagles, and peregrine
falcons, has been related to the abundance of DDT discharges
in this area including the death of birds in the Los Angeles
Zoo attributed to their being fed with fish that were loaded
with DDT and caught in San Pedro Bay. Loss of some 75% of
the wetlands habitat in this area and the fact that many
species do not use this area for nesting but over winter
here or pass through the area on a seasonal basis makes
information on the abundance of birds interesting but so
highly variable that it is diffic- to interpret its sig-
nificance .
c r.
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IIC 135 - "no large blooms" of dinoflagellates "have
occurred since 1974". This coincides with the period
in which massive discharges of DDT into local waters were
abated. It has been suggested that there is a relationship
between phytoplankton populations structure and the presence
of chlorinated hydrocarbons and the population structure
of zooplankton and the presence of heavy metals. The
discharge of heavy metals has also been reduced in this
area with obvious long-term ecological impact still to
be determined.
IID 3 - They report that in the fall, Oct. - Dec. of 1978,
A. tonsa did not participate in the expected bloom near
the outfall, but did bloom at more distant stations. The
start up of 2° treatment is suggested as a cause for this
failure, but other possible changes in physical and/or
chemical parameters (effects of runoff on temp., salinity,
turbidity in shallow vs. deeper water, etc.) are not men-
tioned.
HE 3 and IIE 10 - They report a decline in the total
abundance of benthic animals since 1974 and a steep decline
in diversity in 197 8 (after going to 2° treatment). But
those species which declined in abundance are those which
are tolerant of a primary effluent habitat, and
dependent upon effluent. As the habitat becomes less
impacted, and as recruitment (planting of animals?)
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takes place diversity and abundance less tolerant to stress
of species will increase.
IIF - They report that anchovy populations in the harbor
have declined much more than offshore populations, and
suggest that going to 2° treatment accounts for this greater
decline. But offshore populations were determined acoustic-
ally and this data is not reliable.
They report a decline in the white croaker populations
since diversion of cannery wastes to TITP and going to 2°
treatment. Good - look for greater diversity, not the
restoration of former white croaker populations.
IIF 11 - They note very high numbers of Genyonemus eggs
and larvae in Jan. - Feb. 1978 and suggest that a decline
in possible predators may account for this. They do not
mention that the 30 fold decline in bacteria in the harbor
after going to 2° treatment, plus the improved quality of
the discharge may have had a positive effect on survival of
fish eggs.
IIIA4 - The 10 fold increase in the bacteria population
following the TITP breakdown during the summer of 197 8 is
a good argument for continued 2° treatment.
IIIB 18-19 They conclude that an ecosystem which is

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bacterially enriched and poor in phytoplankton may be as
productive as phytoplankton based ecosystems. But what
is the relative value of these two types of production?
I'm sure that the phytoplankton can support a much more
diverse food web than bacteria.
IIC 5 - They conclude that cell numbers and chlorophyll a
values are probably not directly correlated. This has long
been recognized, and should have come as no surprise to
them.
HID 8 - Inorganic phosphate has never been shown to be
limiting to phytoplankton production in Southern California.
They report that the harbor has not been considered nutrient
limited in the past. They do not discuss why the standing
crops of higher invertebrates and fish are so low.
IVB5 - They report that the benthos, phytoplankton, zooplank-
ton and fish inside the harbor were all lower than would be
expected in April 1978. They mention rain and abnormal
temperatures as a possible cause, but not the TIPT break-
down, and the use of chlorination starting March 9, 197 8,
and how this might have affected life in the harbor.
VB 1 - The choice of Neathes arenaceodentata and Fundulus
is poor. They are not appropriately sensitive species, to
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bioassay the TITP effluent.
VB 4 - The experimental technique used to bioassay the
TITP effluent was very poor. The more tolerant species
lived under all test conditions, while the more sensitive
species showed high mortalities under all test conditions,
including controls. The results appear to be meaningless.
To conclude that no toxic wastes were present in the effluent
tested was presumptuous.
VC 2 - They imply that, because anchovies, in a 15 day
test, showed some weight gain from being fed cannery sludge,
the sludge should be disposed of at sea. Long-term effects
upon the fish, and especially the habitat (promoting a
detritus/bacteria based food web at the expense of phyto-
plankton based web) are ignored.
VD 1 - Mytilus is used to determine the effects of the
waste plume on invertebrates. This is a poor choice;
the animal is tolerant of polluted conditions. It is
not an appropriately sensitve species.
VD 5 - They emphasize that the TITP upset in the summer of
'78 coincided with the faster growth rate in the mussels
nearest the outfall. Why not emphasize that growth rates
CO
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in the spring and winter were reported to be greatest
furthest from the outfall?
VD 6 - They select a station inside the harbor (A2) as a
control station for their biostimulation of inverts study*
This is a poor choice for a location for a control site.
VD 7 - Why was a grocery scale used to determine biomass
change?
VD 9 - They conclude that the TITP effluent is providing
nutrients to the food chain, as detected using settling
plates, but they ignore the point that the station furthest
from the outfall showed the greatest increase in biomass,
indicating a negative effect on plates proximate to the
effluent outfall.
VD 11-12 They report that Mercenaria fed cannery wastes
suffered negligible mortalities, and conclude that the
treatment was not detrimental. This conclusion is not
justified. Sublethal effects were not studied, except for
growth, and growth was reported to be insignificant.
VD 12 - Fundulus is used in an attempt to demonstrate a
bioenhancement effect of cannery waste. Fundulus is a poor
choice. It is not an appropriately sensitive species.
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e
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The controls grew over twice as much as the cannery waste
treated fish. This was determined by them to be statis-
tically insignificant.
They conclude that, based upon their results with Mercenaria
and Fundulus, cannery wastes could have a positive biostimu-
latory effect in the harbor. This is presumptuous. Growth
in Mercenaria was not significant. Growth in Fundulus was
slower in the cannery waste treated group than in the controls.
No other sublethal effects were studied, and the effects of
the discharge upon the general ecology of the harbor (stimu-
lating a detritus/bacteria based food web at the expense of
a phytoplankton based web) are ignored.
VD 13 - They report that phytoplankton removes ammonia from
TITP effluent, and conclude that this suggests optional treat-
ment modes as well as natural biological processes in the
harbor. But it should be emphasized that this applies only
to ammonia not to the other constituents of the effluent.
We have observed, and as Stephens is quoted, a general
decline in the abundances of fishes in the inshore area of
Southern Calfornia in recent years. Our data is consistent
with the data reported by Soule et. al. but based upon
observations made in Santa Monica Bay more than 15 miles
from the TIPT outfall and presumably independent of that
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site of discharged wastes.
Also, we have observed a decline in the abundance and
diversity of certain invertebrate groups in Los Angeles
Harbor, e.g. , nudibranchs, at sites remote from the TIPT out-
fall. These observations on fishes and nudibraphs might be
worked up and presented in a semi-quantitative manner if the
information would be of importance in evaluating the report
of Soule et. al.
As noted in our previous communication to you, Soule
et. al fail to establish a firm basis for the types of inves-
tigations which they conduct related to the impact or lack
of impact of the waste field which is the subject under con-
sideration. Until this is done, it becomes very difficult
to appraise the significance of their results and the con-
clusions stated remain questionable.
Some criteria are suggested below for your considera-
tion in appraising the quality of inshore waters in Southern
California to form a basis for investigations of the impacts
of pollutants.
Standardization of methods is essential and collection
of repetitive observations of key critical qualities or
components is essential. The EPA guidelines for bioassays
are certainly a good example of what may be done in this
respect.
With regard to background environmental data this
should include: temperature, salinity, dissolved oxygen,
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pH, and transparency through the water column. These
observations will at least permit comparisons of water masses
and phytoplankton production or productivity without deter-
mination of inorganic nutrients, e.g., nitrate, ammonia,
phosphate, and silicate.
Characterization of the bottom sediments for organic
content and sediment composition.
Parallel observations for the same components, proper-
ties, biological relationships in an area relatively free of
waste discharges {a control area) on a synoptic basis.
Determination of the concentration of the waste field
at the respective sampling points in the area of the waste
discharge.
Determination of the composition of the waste being
discharged and the variation in this composition during the
period of study.
Experimental observation of the impacts of wastes
upon species indicated to be sensitive to these wastes.
Measurements may include survival, rate of growth, fecun-
dity, morphology, larval development, accumulation of
toxins, behavior, etc. Suitable species would include those
which were known to once live in the area affected or which
are known to exist in similar but presently unpolluted
habitats.
Independent peer review of the proposed investigations
and of the reports of the results of the studies. . This
should be a part of any contract written for work of this nature
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I have been attempting to develop a trophic analysis
argument which involves describing the diversity and abun-
dance of organisms which may occur at a given location in
the absence of pollution. This hypothetical community
wciuld be considered to be that of a balanced endiginous
biota. It would be developed based upon historical know-
ledge of what was known to live in a given location before
it became polluted, and on the basis of what is known to
survive now in comparable areas relatively free of pollutants.
There is a growing abundance of evidence that this approach
has good empirical support and it provides a strong approach
for coastal management. It may well be a more reasonable
way to look at the problem of the harbor rather than to
attempt to manage this area as a waste oxidation lagoon.
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Suite 541
Lo» Angelas, California 90014
Telephone (213) 687-4030
jSZiZSi	C32K21
TT.S3
July 11, 1979
Mr. William MacCleish
Bureau of Engineering
City of Los Angeles
638 So. Beacon St.
San Pedro, CA 90 731
Dear Mr. MacCleish:
Thank you for forwarding a copy of the Draft Environmental Impact
Report on the Terminal Island Treatment Plant (TITP) Unit II-C,
Harbor Outfall. We also obtained a copy of the Harbors Environ-
mental Projects companion study on the Ecological Changes in
Outer Los Angeles-Long Beach Harbors Following Initiation of
Secondary Waste Treatment and Cessation of Fish Cannery Waste
Effluent. I have reviewed these documents with considerable
interest. However, because of my continuing interest in the
development of new facilities and the proper management of
biological resources within San Pedro Bay, analysis of the above
documents precipitated the need to review several other documents
and certain correspondence associated with the evaluation of the
TITP Effluent Outfall and the continuing controversy over the
issue of bioenhancement in San Pedro Bay.
The comments which follow are based upon a review of the documents
listed at the end of this letter. For .purposes of discussion
herein these documents/letters are noted by a reference number.
Based on a review of the concerns expressed in references 8,
9, 10 and 11, it is very difficult to sort out scientific fact
from conjecture, belief, and varying interpretations of regulatory
processes. I recognize that this problem is not new, for while
serving as the Director of Commerce, Port of Long Beach, and as
a member of the South Coast Regional Coastal Commission, incon-
sistencies in the interpretation of data and intent of proposed
actions in San Pedro Bay occurred frequently. All of this is
indicative of the absence of a carefully "orchestrated" scientific
and regulatory analysis of the character and quality of San Pedro
ENVIRONMENTAL FEASIBILITY STUDIES
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Bay and the evolution of planning options based on the continued
use of San Pedro Day as one of the major port areas on the west
coast of the United States.
It is quite clear that San Pedro Bay has been substantially altered
in an effort to develop an efficient and effective maritime center
for the southern part of the west coast of the United States. For
example/ in 1954, the Los Angeles and Long Beach Harbor areas reached
a very low environmental status when the dissolved oxygen content
plunged to 1.0 ppm or less. Substantial efforts were made to alter
mans influence within San Pedro Bay in such a way as to maximize
the opportunity for maintaining an "appropriate" mix of marine
communities. It was this effort which has lead to the present
status of regulatory confusion.
Procedurally, concerns about the TITP outfall should not include
the issue of bioenhancement. However, the Harbors Environmental
Projects docunent (3) obviously has kindled a resurgence of dis-
agreement over the issue of bioenhancement.. The South Coast Regional
Coastal Commission approved the construction of the TITP several
years ago in accordance with EPA requirements. Construction of
the outfall only represents another component of that basic project.
Bioenhancement, whatever it may be, is related to the general
discharge of effluent into San Pedro Bay, not with the specific
relocation of the outfall.
My review of references 1 and 3 and references 8, 9, 10 and 11
indicates that many of the concerns center around the issue of
whether or not one set of data are more credible as compared to
another and whether or not a given approach is more appropriate
(adequate) than another and whether the "belief" of one scientist
or regulatory agency staff member is more significant than that
of another. For example in the letter from the Scripps Institute
of Oceanography (11) it is noted that "three of the six comparative
studies sampling strategies were altered between the earlier and
later studies.... and that this undermines the strength of the
results." Indeed this is the case, i.e., the sampling strategies
were altered for the zooplankton and the fish egg sampling, but
this was done as a consequence of interpretation of the data
collected at the beginning of the comparative studies. Yet, it
seems clear from the information in reference 3 that continued
sampling, without modification, would have resulted in essentially
negative input, i.e., the desired organisms would not have been
captured. Another illustration is the comment in the Fish and Game
letter of Hay 22, 1979, (9) where the statement is nade that "we
believe that marine ecosystem management programs incorporating
inputs of wastewater as a major component of the plan should be
confined to closed, carefully controlled systems such as hatcheries
Reproduced from
best available copy.
2
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or, at the most liberal extreme, in marshes created or augmented
by carefully controlled wastewater flows." Such a statement indicates
a predetermination that managed release of wastewater into any other
type of marine environment is harmful. To the best of my knowledge,
there are no data substantiating this belief. In the same letter (9)
a reference is made to the omission in the Harbors Environmental
Projects study (3) of any meaningful comparisons with other similar (?)
enclosed bays such as San Diego Bay. Although this criticism may be
appropriate for gross comparisons, it has little merit in this
instance since an adequate long term analysis of the physical and
biological characteristics of San Diego Bay has not been achieved.
Therefore, to assume that San Diego Bay has been rejuvenated (9)
because certain "indicators" have adjusted, is to grossly over-
simplify. Further, in the State Water Resources Control
Board, Division of Uater Quality letter of June 13, 1979 (8) it is
noted that the information on fish fauna in reference 3 "seems to
be based entirely on the results of Harbors Environmental Projects
surveys" and further questions why "results of other studies, such
as those performed for Southern California Edison Company" (4) were
not included for purposes of comparison. A cursory review indicates
that the statistical approach used in the Southern California Edison
Company report represents what can be generally called the "smoothing"
of statistics, i.e., the "lumping" of three-four years of data. For
purposes of the analysis in the Harbors Environmental Projects document
such lumping was not appropriate. This appears to be a good illustra-
tion of the interpretation prerrogative assumed by various individual
scientists, regulators, etc.
The above illustrates that a "reasonable" concensus must be evolved
in order to insure "maintenance" of the marine environment as well
as efficient operation of existing and proposed facilities within
San Pedro Bay. Such a goal of compatibility is not unreasonable,
but it can not be achieved if there is continued "agitation" due
to the absence of clearly developed regulatory guidelines, including
essential definitions such as for enhancement, and a common goal of
effectively analyzing problems usinq available data. Assuming that
this is the goal of all the studies referenced herein, then the con-
cern should not continue to center around absolute compatibility of
sampling strategies or whether the white croaker is being sold as
a "butterfish" or whether the failure of the kelp bed to grow on the
middle breakwater should be linked to toxicity from the TITP effluent
(until adequate data disclaiming same are available) or whether the
TITP effluent is the only remaining nutrient point (or non-point)
source in the harbor, and so forth. Rather, the approach should be
changed to "how can we determine the best possible mix of activities
in San Pedro Bay which will yield a rich and diversified assemblage
of organisms (within an "appropriate" food web), regardless of

3
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whether they are unique to San Pedro Day, comparable to those in
the California 3ight or parallel to certain assemblages in Newport
Bay, San Diego Bay, Monterey Bay, etc.
In order to get this issue off dead center, it is imperative that
the following be accomplished; until these are accomplished, the
continuing debate over bioenhancenent and biological parameters
of San Pedro Day will continue unabated and unresolved:
•	What is bioenhancement (enhancement)?
Most of the references differ on whether bioenhancement has been
demonstrated. Yet for all intents and purposes, there is no working
definition of bioenhancement offered by the EPA, the State Water
Resources Board, the Regional Water Quality Control Board, the
Department of Fish and Game or the Fish and wildlife Service, etc.
If these regulatory agencies are to participate effectively in
determining when bioenhancement does or does not occur, they must
promulgate a usable definition. To criticize an analysis without
the availability of a basis for comparison is to be remiss.
•	Development of a task force and/or single
agency responsible for making decisions
on bioenhancement.
When EPA delegated to the State of California Water Quality Control
Board and the regional boards the responsibility to make deter-
minations and issue permits for effluents, it was assumed that their
decision on bioenhancement would be acceptable. Yet, history
indicates that this was not the case with respect to cannery wastes
or TITP effluent. During all this discourse, no definition of
bioenhancement was provided, and there was no "concensus" approach
between the various federal and state agencies on how to resolve
the interpretation dilemma. With the opportunity for evolution of
as many definitions as participating decision-makers, resolution
of the issue almost is impossible.
o "Normal" environment within San Pedro Bay.
Until a decision evolves which indicates what is acceptable and
within what limits, with respect to basic biological components
in the environs of San Pedro Bay, additional studies only will be
that, just additional studies. The regulatory process must provide
clear guidelines for all to use. Such guidelines must have some
flexibility. These guidelines are essential for the continued
survival of San Pedro Bay. In essence, we must translate our past
totally human oriented goals into goals which maximize both h\iman
desire and environmental stability; we must establish a "normal"
environment definition.
Reproduced from
best available copy.
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• The desirability of an empirical study.
The individual, geographically specific studies conducted over the
last few years within San Pedro Bay have been focal points for
criticism because they have not been conducted with "sufficient"
complementarity, i.e., sampling strategies, statistical methodology,
key assumptions and so forth have been at variance. Yet, without a
large source of investigative funds, and agreement as to the
methodology and approach between all agencies having ministerial
and discretionary approval, no such empirical study will occur.
The likelihood of such a study occurring in the immediate future
is nil. Therefore, we are left with the series of individual
studies, representing an imperfect data base, but from which we must
make the best possible interpretations in order to manage the
resources of Han Pedro Bay.
In conclusion, at the expense of being as autocratic as those I
have criticized above, the available data support the concept
of bioenhancement with respect to the introduction of some cannery
effluent and some TITP effluent into Gan Pedro Bay. Mo data have
been presented to show that bioenhancement has not occurred!
Thus, without the opportunity to conduct the empirical study
noted above, and if we are to proceed with any kind of appropriate
management within San Pedro Bay, i.e., not just say "no" to any
change, we must proceed with "managed" introduction of certain types
and certain volumes of effluent into Gan Pedro Bay. To allow
individual agencies, etc. to continue to express their "beliefs"
without adequate supportive information is to defy the basic tenents
of scientific analysis.
Thank you for this opportunity to comment on these documents.
I hope I have been constructive, and I hope that you too will
persevere in achieving the managed use of effluent in the San Pedro
Bay whereby we can clearly demonstrate the degree and character
of the related bioenhancement process.
Sincerely,
5
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Title
The Draft Environmental Impact Report,
Terminal Island Treatment Plant Unit II-C,
Harbor Outfall.
Draft Environmental Impact Report, Terminal
Island Treatment Plan Unit II-C, Effluent
Disposal System.
Ecological Changes in Outer Los Angeles-Long
Beach Harbors Following Initiation of Secondary
Waste Treatment and Cessation of Fish Cannery
Waste Effluent, Harbors Environmental Projects,
University of Southern California (April, 1979).
Marine Monitoring Studies, Long Reach Generating
Station, Southern California Edison Company
Preoperational Report, 1974-1976, prepared by
Environmental Quality Analysts and Marine Bio-
logical Consultants, Inc. (June 1977).
Environmental Investigations and Analyses Los
Angeles-Long Beach Harbors, 1973-1976, Final Report
to the United States Army Corps of Engineers
prepared by Harbors Environmental Projects,
University of Southern California (December, 1976).
Draft Environmental Impact Report, Master Environ-
mental Setting, Volumes 1 and 2, Port of Long Beach,
prepared by Soils International, Allan Hancock
Foundation, and Socio-Economic Systems, Inc. (1976).
Summary of Knowledge of the Southern California
Coastal Zone and Offshore Areas, Vol\imes I-III,
prepared by the Southern California Ocean Studies
Consortium (1974).
State Water Resources Control Board, Division of
Water Quality, Letter to Mr. L. Frank Goodson,
Project Coordinator, Resources Agency, Sacramento,
California, dated June 13, 1979, signed by Neil Dunham,
Division Chief, Manager-Clean Water Grant Program.
Letter from Department of Fish and Game, Marine
Resources Region, Long Beach, California to
Mr. Jeffrey D. Denit» Chief, Food Industry Branch,
EPA, dated May 22, 19 79, signed by Robert G. Kaneen,
Regional Manager.
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Letter from Department of Fish and Cjar.e to
L. Frank Goodso.n, Project Coordinator, Resources
Agency, dated June 7, 1^79, signed by E. C. Fullerton,
Director.
Letter from Scripps Institution of Oceanography
to Mr. Anthony V. Nizetich, Director, Government
Relations, Star-Kist Foods Co., Inc., Terminal
Island, California, dated August 20, 1979
by E. L. Venrick.
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|e rn o r a n d u m
• 1. L. Frank Coodsori, Projects Coordinator	Dote: June 7, 1979
}	Resources Agency
2. A-95 Coordinator
State Water Resources Control Board
I>. 0. Box 100
|	Sacramento, California 95801
n j Department of Fish and Game
ject: SCH 79051509A — DEIR Terminal Island Treatment Plant Unit II C Effluent
Disposal System and Harbor Outfall
Af ter~ reviewing the subject documents and supporting da'ta, the Department
of Fish and Game believes that the City of Los Angeles has not demonstrated
that Terminal Island Treatment Plant (TIT?) effluent enhances the receiving
vaters of Outer Los Angeles Harbor and therefore in our opinion the alterna-
tive of ocean discharge is preferable to discharge to the harbor.
The following material represents our comments on the three documents sub-
mitted for review- One of the documents, entitled Ecological Changes in
Outer Los Anke1es-Long Keach Harbors Following Ini t ia t i on of Secondary
Waste Treatment and Cessation of Fish Canr.erv Waste El'fluor.t was prepared by
the Harbors' Environmental Project (HE?) of the Allan Hancock Foundation.
This document was submitted by the City to support its contention that
wastes discharged from the TITP enhance the receiving waters. We conducted
an earlier review of the HLP document, rit the request of the Environmental
Protection Agency, and have attached those comments as an integral part of
this review.
The remainder of the following connents will focus primarily upon the
documents concerned with the Harbor Outfall cind the Effluent Disposal System.
As an introductory comment, we note that this pair of documents is presented
as Phase I of the Effluent Disposal System by the City of Los Angeles. The
City is attempting to demonstrate that effluent from the TIT? provides en-
hancement of the receiving water—i.e. enhancement as defined in the State
Water Resources Control Board's (SWRCB) Water Quality Policy for the Enclosed
Eays and Estuaries of California (The Bays and Estuaries Policy). If the
City's attempt is ratified by the Regional Water Quality Control Eoard (RWQCB)
and/or SWRCB then the Harbor Outfall project could represent the completed
Effluent Disposal System project;'Otherwise an ocean outfall would be in-
cluded in the project.
Because the Department of Fish and Cane believes enhancement of harbor waters
has not been demonstrated, we regard the Harbor Outfall as an interim project.
Even though the documents are presented as two separate DEIRs, we think the
Outfall document only discusses a temporary solution to problems related to
development by the Port of Los Angeles, and would not, therefore, be a com-
pleted project.
For the above reason and because many sections of the two documents are
identical, the comments that follow are offered for both documents but are
keyed to the pagination of the more comprehensive Effluent Disposal System
|jocument, unless otherwise noted. 7Reproduced from	- o
j best available copy.	- -•
RESPONSES OF HARBDRS ENVIRONMENTAL
PROJECTS, USC, TO DEPARTMENT OF
FISH AND GAME MEMO DATED JUNE 7, 1979.
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RESPONSES BY HARBORS ENVIRONMENTAL PROJECTS,
UNIVERSITY OF SOUTHERN CALIFORNIA, TO STATE DEPARTMENT OF FISH AND GAME
REVIEW OF DRAFT EIRs FOR CITY OF LOS ANGELES, TERMINAL ISLAND
TREATMENT PLANT, dated June 7, 1979
GENERAL COMMENTS
1. The Effluent Disposal System document is inadequate because it contains only
a limited discussion of but one other project contemplated for the Los
Angeles-Long Beach Harbor complex. The Harbor Outfall document lacks such
a discussion. We believe the documents should more completely portray
future projects (such as detailed in available port master plans and local
coastal planning documents) under consideration by the City of Los Angeles,
Port of Los Angeles, City of Long Beach, Port of Long Beach, or others.
The documents should especially focus on the cumulative impacts of projects
and impacts that could alter flushing patterns for harbor v/aters. We also
think the documents should particularly discuss the adverse effects that
altered or restricted flushing patterns may have upon ambient water quality
and upon the enhancement effects alleged by the City for TITP effluent dis-
charged to the harbor.
Response: Projects that can be assessed in terms of cumulative impacts require
sufficient description of those projects to permit reasonable extrap-
olation of impacts. With the exception of the first phase of Los
Angeles Harbor deepening and land fill, no such description exists
for projects that would interact significantly with this one. The
Port of Los Angeles does not have a Master Plan that has been accepted
by the Coastal Commission. The Port of Long Beach Master Plan, accepted
by the Coastal Commission, has no central outer harbor fill projects.
The reverse consideration is perhaps more appropriate, that as future
projects are developed they should be considered in light of their
cumulative impacts on the present project,if it becomes a reality.
Documents cited in the DETR include the Allan Hancock Foundation (1976)
report to the Army Corps of Engineers, which discussed the impacts
of all proposed fill projects on both Ports. Also cited was McAnally
(1975) and other reports of the Army Engineers on tidal flushing and
the effects of fill configurations.
2. In Section IV.C.l. both documents discuss plant malfunctions ("upsets") as
unavoidable adverse impacts, but of a type that can usually be "quickly"
brouqht under control. Predicted adverse effects are attributed to chlorine
that will presumably be added to control bacterial contamination caused by
the discharge of incompletely treated effluent. We believe adverse impacts
during upsets will not be limited to added chlorine. Instead, we believe
chlorine addition will significantly compound severe adverse effects that
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-2-
are often caused by untreated industrial wastes discharaed during upsets
regardless of where the discharge would occur. We further believe adverse
effects of an upset will be compounded if discharged to the harbor because,
according to the document (Section 111.0.4.6.(2) Circulation and Flushing),
a maximum of 19 percent of the waters in the outer harbor is "tidally dis-
placed"at a mean tide of 5.4 feet. Although the term "tidally displaced" is
unclear, as used in the document, we think it indicates a significant
retention time for wastes discharged to the harbor. This suggests that
for each day that an upset is in progress, concentrations of improperly
treated waste will increase proportionately and, further, that the adverse
effects will linger for several days after an uoset is corrected. We
believe the adverse effects of an upset should be more thoroughly discussed
especially with reference to the "full and uninterrupted protection of
beneficial uses" clause contained in the Bays and Estuaries Policy.
We believe that upsets will cause more harm to living resources if effluent
is discharged to the harbor than would occur if discharged to the open
ocean because the open ocean has greater potential for raDid and continued
dilution than the harbor with its potentials for significant retention
time in concert with comparatively reduced dilution.
Response: Section IV.C.l does not state that	plant malfunctions ('upsets')
... can usually be 'quickly' brought under control". No attempt was
made to minimize the potential impacts or to claim that chlorinazion
alone would alleviate impacts. The statement, as originally written,
agrees with most of the DFG views, with regard to chlorination.
Since the section referred to was originally written, a major plant
upset did in fact occur in the summer of 1978. Data were collected
documenting both biological and other environmental conditions
before, during, and after the upset. These data are reported on and
discussed in detail in Soule and Oguri (1979),
The findings of these investigations indicate that both DFG, in their
comments, and the authors of that section of the document were wrong.
The upset led to increases in fish caught by trawl in the area and
increases in numbers of benthic organisms and numbers of species found
in the area. Because the major industrial wastes are biodegradable,
any negative effects were apparently transitory. Possible effects on
zooplankton and benthic organisms were fully discussed in Marine Studies
of San Pedro Bay, California, Part 16, incorporated by reference
(Soule and Oguri, 1979).
Impacts of ocean disposal are not eliminated by dilution; when large
quantities of fresh water are introduced into an entirely oceanic
environment, many stenohaline species will be eliminated from the
immediate receiving waters.
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SPECIFIC COMMENTS
Page 1-3 (Harbor Outfall document) - The document states that "TITP effluent is
theonly remaining nutrient source to the harbor..." This statement
is inaccurate. Because harbor waters are constantly exchanged with
waters from the open ocean, nutrient input is also constant from that
source. Also, intermittant input occurs from periodic runoff during
rainstorms. The statement should be revised to say that TITP effluent
is the only remaining point source for nutrients (i.e., of human
origin) and the document should further reflect natural and nonpoint
source inDuts that occur as well.
Response: Within the context used, point-source was the intended meaning. TITP
became the only year-around, somewhat uniform nutrient source, with the
elimination of cannery outfalls. In contrast, the storm drain and river
flows are highly variable, and also flush hydrocarbons, metals, fertili-
zers and other urban chemicals into the harbor. Some of these sub-
stances are biodegradable by bacteria and some are biostimulatory to
phytoplankton, but these non-point source nutrients alone may not sus-
tain a healthy ecosystem. The data on mineralized nutrients show that
levels outside the harbor are much lower than in the harbor.
Nutrient input into the harbor waters from TITP considerably outweighs
that from all other sources in terms of quantity of both organic and
inorganic nutrients.
Page 11-9 - In a di.scussi.on of the Consolidated Slip Alternative the document
states it hgs not been demonstrated that enhancement of the receiving
waters would result from continual discharge of variable quality
effluent from TITP. The discussion should clearly state why enhance-
ment that is claimed to result elsewhere in the harbor from dis-
charges of TITP effluent could not be demonstrated for this alterna-
tive.
Response: Enhancement in the outer harbor is predicated upon the assimilation
capacity of the receiving waters. Consolidated Slip does not have
an adequate water volume to serve as receiving waters, and the nodal
point of the tides is nearby so that flushing is very poor. The
physical configuration of the inner harbor, with its many dead-end
slips, also tends to promote poor flushing and isolation of each small
area from the others. Enhancement in the inner harbor was therefore
felt to be less likely than for the outer harbor.
Page 11-10 The document should explain why trace metal contaminants and biologi-
cal vectors present in TITP effluent are a negative aspect for the
Harbor Lake alternative but not for the Outer Harbor Outfall alterna-
tive. We think that metal contaminants and biological vectors signal
adverse impacts for living resources (including mankind) regardless
of the alternative selected but that they would have less such effects
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if an ocean disposal alternative were implemented.
>
Response: The EIR did not specifically address "trace metal contaminants and
biological vectors" but in Section II.C.6. cited other problems of
a more general and inclusive nature.
Variability in the salinity of the TITP effluent and its other char-
acteristics were discussed. The ability of Harbor Lake to cope
with a plant upset is greatly limited by the small volume of water.
For these and other reasons the Harbor Lake alternative was rejected.
It should be noted that if "biological vectors" refers to the possi-
bility of public health impacts,then the outer harbor, with its
higher salinity, greater volume and more rapid dilution would be
safer for receipt of the effluent. In any case there have been no
public health problems associated with the use of secondary treated
waste water for golf course irrigation. Such use is made of waste
waters by the Moulton-Niguel Water District, the Laguna Hills
Sanitary District and the City of San Clemente, to cite just a few.
There have been no public health problems identified in the outer harbor
receiving waters.
Page III-l The document states that the proposed 30 million gallon per day flow
from TITP represents less than 3 percent of the total effluent input
to the southern California coast. We believe the TITP effluent should
be more meaningfully compared to waterflow into the harbor.
Response: Data based on hydraulic model studies conducted by the U.S. Army
Engineer Waterways Experiment Station (McAnally, 1975) showed that
the two flood phases of a mean tide of 5.4 feet would result in
8280 X 106 cubic feet of water entering the harbor through Angels
Gate, Queens Gate and the eastern entry into the harbors. If only
Angels Gate is considered, the values from the hydraulic model data
show 3550 X 106 cubic feet of water entering the harbor. Data based
on prototype (actual field) measurements show the tidal volume
entering the harbor to be 3100 X I06 cubic feet for the two flood
phases.
The 30 X 106 mgd stated as the design flow for TITP converts to
31.25 X 106 gallons for 25 hours, which is closer to the time in-
volved in a complete cycle of two high and two low tides that normally
occur in the harbor. This, based on 231 cubic inches per gallon,
converts to 4.18 X 106 cubic feet.
Consideration of tidal flux alone in calculating the desired data
skews the values upward, giving an exaggerated view of the quantity
of effluent as a percentage of total flow. However, presented as a
"worst case" situation it gives some basis of comparison to the
"less than 3% of the total effluent input to the Southern California
coast." The data are tabulated below.
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-5-
Data base
TITP Effluent Volume
Tidal Prism Volume ^
Model data
All harbor entries
0.05%
Angels Gate
0.12%
Prototype data
Angels Gate
0.13%
McAnally, W.H., Jr. 1975. Los Angeles and Long Beach Harbors Model
Study. Report 5. Tidal verification and base circulation tests.
Technical Report H-75-4. U.S. Army Engineer Waterways Experi-
ment Station, Vicksburg, Miss.
Page II1-7 The composition of TITP is described as 30 percent domestic waste
and 70 percent industrial. The chemical constituents of industrial
discharges should be portrayed and expected discharges of those
constituents during plant malfunctions should be portrayed under
"worst case" conditions.
Response: The industrial portion of the wastes treated by TITP are primarily
wastes from the fish canneries. These are food processing wastes
and contain no exotic chemicals or unusual concentrations of heavy
metals or chlorinated hydrocarbons. The most unusual characteristic
of this material is the high level of BOD. This is reduced consider-
ably by the secondary treatment process.
The chemical constituents at both the influent and the effluent
during normal operation and upsets are shown in the 1978 annual
report to the Regional Water Quality Control Board where it is
available as a document of public record.
Page II1-28 SII1-29 - The flushing rates and circulation patterns for the outer
harbor should be expressed in numbers of days to achieve complete
flushing. This information is needed to more clearly understand
the effects of plant malfunctions listed as "unavoidable impacts"
elsewhere in the document.
Response: Assuming a 19% water exchange (max) per tidal cycle, approximately
ten days are required for a 99% exchange of water. This can be
considered complete flushing. About 80% flushing would occur in
four days with two tidal cycles per day. For consideration of the
significance of this, please refer to the response above to the
comment on Page III-l of the EIR.

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-6-
Page 111-38 The document states that restrictions of cannery wastes caused
"... a loss of nutrients to sustain the biota." This statement
contradicts the statement on page 111-32 which says "In the Harbor,
nutrients are rarely limiting but inhibitory (excess) amounts may
also occur." We believe the latter statement is valid because
(1) the harbor exchanges waters (containing nutrients) with the
open ocean and (2) waste discharges to the harbor not only provide
maintenance for artificially high populations of some animals,
with variously perceived values, but also bear the continuous
potential to cause damaging excesses (e.g., treatment plant
malfunctions).
Response: The confusion arises from taking the two comments out of context,
or of failing to recognize that mineralized nutrients for phyto-
plankton, such as nitrate and phosphate (discussed on pages 111-31
and III-32) are quite different from the complex organic nutrients
for benthic organisms such as the fish cannery wastes (referred to
on page 111-38}.
The statement above regarding the provision of nutrients to main-
tain "... artificially high populations of some animals, with
variously perceived values ..." suggests that the evidence for
bioenhancement is accepted but that the reviewer cannot accept it
as having value. The great diversity of species in the harbor
belies the implication that the ecosystem is somehow not of value.
The further allusion to "... damaging excesses (e.g., treatment plant
malfunctions)," suggests that the reviewer has not reviewed the
information supplied on the effects of the treatment plant mal-
function during the summer of 1978, when benthic organisms, fishes
and birds increased in numbers and in numbers of species.
Page IV-9, 10 and Page IV-22, 23 - Statements on these pages discuss the effects
of Phase II landfills proposed by the Port of Los Angeles. The
document lacks, however, a portrayal of other similar projects con-
templated for the harbor. As we stated in our general comments,
these projects should all be discussed with regard to the effect
they would have on flushing patterns in the harbor and how any
alteration in flushing patterns they may cause would affect the
enhancement issue.
Response: There is no legal requirement nor logical justification for address-
ing, as cumulative impacts, the influence of projects that are not
yet designed or authorized. Phase II landfills were addressed in
the Dt'IR, although it is already apparent that the descriptions
available prior to preparation of this EIR have now been con-
siderably modified. If the outfall is built prior to future project
initiation, the future project should bear the burden of evaluating
the cumulative impacts.

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-7-
The Port of Long Beach Master Plan does not include any central
Outer Harbor fill. The Pier J completion project was tested by
the Army Engineers Waterways Experiment Station and found to
incur only localized circulation impacts. The Oil Terminal (SOHIO)
vas considered to have a net biological enhancement. Both the Port
of Long Beach Master Plan and the Oil Terminal were approved by the
Coastal Commission.
Page IV-28; Page V-9 - The document states that the East Ocean Outfall alterna-
tive would have greater impacts on marine biota in the open ocean
than the harbor outfall alternative would have on biota in the
harbor and that the discharge would adversely affect recreational
beaches at Cabrillo Beach and the nearby marine life refuge. We
disagree. We believe the waste discharge requirements can and
would be set to protect the beneficial uses of the receiving waters
(which include recreational values and protection of living
resources) by the RWQCB in accordance with PL 92-500 and with the
Water Quality Control Plan for Ocean Waters of California.
We believe, in the case at hand, ocean discharge is preferable to
harbor discharge because dilution would be more rapid, because
the effects of malfunctions would be less, recovery would be more
rapid, and because contrary to the opinion expressed in the docu-
ment, ecosystems in enclosed bays and estuaries are usually more
sensitive to waste discharge than the ecosystem of the open ocean.
Therefore, we believe the statement and opinions in these pages
regarding relative sensitivities of ecosystems should be docu-
mented, modified, or deleted.
.ResponseThe harbor biota has been exposed to the waste discharges in the
harbor for many years and has adjusted to their presence. In the
past we have noted that even within a species there is a gradation
of tolerance to stress, such as is found in the harbor.
The East Ocean Outfall alternative introduces a stress to an envi-
ronment that has no history of exposure to it. As stated in the
EIR it is therefore felt that "... the initial iiopact would be
greater ..." and that "... impact may be felt at the recreational
beach, Cabrillo Beach, and the Marine Life Refuge..." (emphasis
added) for this reason.
Waste discharge criteria will^undoubtedly be established to protect
the beneficial uses of the receiving waters, but it is apparent
that the area influenced by other outfalls is extensive and obvious,
although such criteria do exist. It would be difficult to imagine
that these criteria will totally prevent the initial impact that
may eventuate if the East Ocean Outfall is built.
The statements and claims made in paragraph 2 are totally undocu-
mented. Dilution may or may not be more rapid but we would seriously

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-B-
question the statements claiming that effects of plant malfunction
would be less or that recovery would be more rapid. Data docu-
menting the in-harbor effects of a major plant upset and the
recovery of the ecosystem from it were presented in Soule and
Oguri (1979). Similar data were not presented by DFG for such
an occurrence in an open coastal region similar to that projected
for the East Ocean Outfall.
The statement that "... ecosystems in enclosed bays and estuaries
are usually more sensitive to waste discharge than the ecosystem
of the open ocean" (emphasis added) is undocumented. This is pos-
sibly true of bays and estuaries without a history of such dis-
charges. However, the harbor, with its long history of accept-
ing such discharges, might equally be quite sensitive to their
removal and this has been documented.
Page V-l The document states that implementing the East Ocean Outfall alterna-
tive "... would remove the last remaining nutrient source from the
harbor..." This statement is false and should be modified as we
suggested for similar statements elsewhere in the document.
Response: Please refer to the response to the comment on page 1-3 of the EIR.
Soule, D.F. and M. Oguri. 1979. Ecological changes in outer Los Angeles-Long
Beach Harbors following initiation of secondairy waste treatment and cessa-
ation of fish cannery waste effluent. In Marine Studies of San Pedro Bay,
California, Part 16. Allan Hancock Foundation and Sea Grant Program,
Vniv. So. Calif. 737 pp.

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REAR ADMIRAL O.D. WATERS, JR., U.S.N. (RET.)
1260 Cedar Lane
North Indialantic, FL. 32903
Critique on California Department of Fish and Game report to
EPA on Dr. Soule's Ecological Changes in Outer Los Angeles-
Long Beach Harbors Following Initiation of Secondary Waste
Treatment and Cessation of Fish Cannery Waste Effluent.
General. In the first place, one could never accuse this critique
of being objective. In spite of the statement in the forwarding
letter that "we look forward to a continuing dialogue with
Dr. Soule and her colleagues" the general tenor of the report is
destructively critical.
Section IA— Bioenhancement. This section strikes me as another
example in its general context of bureaucratic wordsmanship.
For example: Approximately the first four pages are devoted
to an exercise in semantics as to whether something is a "policy"
or a "plan", the principal difference seeming to be that the
latter contains a "program of implementation". This seems to me
to raise a jurisdictional matter which is a separate considera-
tion and merely beclouds the issue under consideration.
I concur that "enhancement" not only "can and should be"
defined, but that this has been done.
Summary and Conclusions
1.	The preference for hatcheries and aquaculture over "open
aquaculture" overlooks the relative costs involved and the many
disappointing failures to date of pure aquaculture ventures.
2.a.	There is a lack of emphasis in the list of beneficial uses
defined by the Los Angeles RWQCB 1978 Basic Plan on "natural or
biological environment".
2b. Is it necessary and is it possible to make these fine
distinctions?	* [
3.	Again, cost is ignored.
4.	No comment.
5.	So what — so long as overnutrification does not occur. Can-
not this be monitored?
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BIOLOGICAL RESOURCES
Summary of Contents. The frequent use of the words "we believe"
unsupported by technical back-up seriously weakens this whole
section.
Section IIA Fish Populations in L.A./L.B. Harbors
No comment on this section. It is a difference of opinion
on valid methods of measuring populations. The comment seems
to be all "opinion".
Section IIB
Again, "we believe that without data from years between the
1973-74 survey and 1978 survey that trends cannot be accurately
portrayed for bird populations". Why ? ? ?
"Our staff expert believes avian populations in the harbor
area are healthy and stable." Again, Why? ? ? ?
Section IIC, IIP, HE
I have only one question here — Who performed the Southern
California Edison survey and how capable are they, but especially
what is the likely bias of a report by an industry discharging to
the harbor?
Sections IIF, IIIA through E, IVA & B, VA, VP
To offer an obscure definition of the word "stimulate", i.e.,
"to provoke", is really grasping at straws I Incidentally, my
Webster's Unabridged does not offer the synonym "to provoke".
It is true that the Latin root is "stimulus"—a goad— and if
you want to stretch that to provocation, I suppose you can, but
it's a long stretch.
Section VB
No comment.
Section VC
The cost of reclaiming the nutrients as "a potentially
marketable fish food source" is ignored.
*
*¦	N
/ ^ 		^// ' J	W " - F -f	7 \
L' ? i ' t ¦ .* ft v: < < /y.	
Rear Admiral O.D. Waters, ,Jr., U.S.N. (Ret.)
t >
Former Oceanographer of the Navy and
Head, Dept. of Oceanography,
Florida Institute of Technology
Date: ^
• ; >. .f
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SOUTHERN CALIFORNIA COASTAL WATER RESEARCH PROJECT
1500 East Imperial Highway, El Segundo, California 9024 5
(213) 322-3080
13 June 1979
Dr. Dorothy Soule, Director
Harbors Environment Projects
University of Southern
California
Los Angeles, CA 90007
Dear Dorothy:
I have just reviewed Part 16 of the Marine Studies of
San Pedro Bay entitled "Ecological changes in the outer
Los Angeles-Long Beach harbors following ..." Most of the
limited time I could spend went into reading the Executive
Summary, the Table of Contents, and skimming rapidly through
the rest of the book. Then I asked two of our biologists
to do the same. We three agree on the following points:
There was substantial bicenhancement (increase of all
kinds of sea life) when the cannery waste was discharged
directly into the harbor and when the TITP secondary plant
failed. It is hard to see how anyone could disagree with
this finding. Everyone knows that when food is available
animals are attracted—and they, in turn, attract other
animals.
If showing bioenhancement is the entire point of the
study, you have proven it to our satisfaction. However,
¦since the title suggests all kinds of ecological changes
it is worth remembering that each reader finds something
different to look for that relates to his own perceived ob-
jectives. For example, if the object of the treatment is to
"clean up" the harbor by increasing water clarity it might
be regarded as successful (I.can't tell from your data). If
the objective of treatment is to maintain or increase the
sealife, it is a failure.
We feel that there should be some discussion of other
ecological changes besides bioenhancement for the benefit
of persons with other interests. It would also be helpful
-------
if there was a table comparing cannery wastes with TITP
and another which compared the principal parameters during
each of the main conditions of discharge.
Once someone decides whether it is more desirable to
have clear water or more fish.the answer will be evident.
I hope this is helpful. With best wishes,
Sincerely
Willard Bascora
Director
WB :es
-------
hlWi
m

¦JSJOS 3
Star-Kfst Foods, Jnc.
l^asTzU

S,

TERMINAL. ISLAND. CALIFORNIA S073?
September 13, 1979
Cibl* AadfMl "FRENCHSACO*
T«l»xt 65-6342
Amwarback; Sl»r.Kl»t ftrn.
Mr. Cal Dysinger
Environmental Protection Agency
1+01 M Street, S. W.
Washington, D. C. 20^60
RE: Section Seafood Study
Dear Mr. Dysinger:
We should like to offer.some comments and observations on the Harbors Environmental
Projects of the U.S.C. Summary Report issued in February, 1979, relating to eco-
logical changes in Outer Los Angeles and Long Beach Harbors and various reviews
of this document, all of which have been submitted to you for consideration in
the Section Jk Seafood Study. Our remarks are of a general nature and refer in
some cases to matters of policy and the question of bioenhancement with respect
to this report and criticisms of it, as well as to the general question of
bioenhancement with biodegradable non-toxic wastes from fish canneries.
1.	In our judgment, the above-mentioned report demonstrates a sufficient
case for the managed discharge of fish cannery effluent waste to the
Outer Los Angeles Harbor to justify the continuation of such dis-
charges during an extended study period. During this period
quantities of effluent discharge would be managed in accordance with
previously agreed parameters and the environmental impact monitored
according to a program agreed by representatives from relevant
regulatory agencies, scientific bodies and industry. In'the case of
Los Angeles.Harbor, this would be particularly appropriate since
all present indications are that the municipal treatment plant
currently being constructed will not be able to handle the total
volumes of industrial waste and significant further capital expendi-
tures would be required.
2.	We understand that the report was a summary of various separate
studies carried out over a considerable period of time by different
scientific bodies for different purposes and which were.funded from
various regulatory or industrial funds. Although we believe that
some of the adverse criticism'which the report has attracted is
justified on scientific grounds, .(e.g.), because some conclusions
in the Executive Summary were based upon data which-was not strictly
comparable, we believe that for the most part this adverse criticism
is misplaced for the following reasons:. .
a. The critic has not appreciated perhaps the fact that
the report was not the result of a, single experiment
designed specifically to demonstrate bioenhancement
. >
-------
Mr. Cal Dysinger
Page 2
from fish cannery effluent in all the circum-
stances that might conceivably have arisen, and
to a large degree did arise, in the period covered
by the study.
b.	The study represented by the report probably has
cost in total several million dollars. In such
major investigations as this studying environ-
mental effects over a period of years, decisions
have to be made to limit the amount of data
collected on some cost effective judgmental basis.
Since nobody's judgment is perfect in advance in
so complex a situation, it is inevitable that
there vill be some gap in the data which subse-
quently cannot be rectified.
c.	With such an extensive study over this period of
time, it is not practical to believe that an
assembly of all the best experts even vould be
able to forecast accurately every eventuality that
would require inclusion in the experiment design.
d.	Even though individual sets of data may be less
than conclusive or strictly comparable, where
several sets of data indicate similar results,
then conclusions may be drawn with a reasonable
degree of confidence.
We do feel, therefore, that some of the adverse
criticism is ill-judged and unfair and is a result
of the luxury afforded by hindsight.
3- With regard to comments in (2) above, and with the expeience of the
above report in mind, we would suggest that whilst it is incumbent upon
some other person to execute the work, it is reasonable to expect
regulatory agencies to take part in experiment design and by implica-
tion to take some responsibility for the adequacy of this design when
the results are reviewed.
U. We agree with the concept that bioenhancement needs to be proven on a
site specific basis. We believe that in each particular case the final
decision should be made having regard for the environmental impact and
the economic impact of the various alternatives.
5. It is difficult to adequately define the term bioenhancement. But in
the absence of a definition, the question becomes one of judgment.
For example, if there is a substantial increase in species diversity
and population, but a significant reduction in the population of one
particular species, does this represent bioenhancement or not? How is
the proportion to be tested if such a test will not comply with an
existing water quality standard or policy, and who will authorize it?
, / . >
-------
Mr. Cal Dysinger
Page 3	t
Is bioenhancement to be rejected in every case where there is any
zone of inhibition? We believe that it is this sort of difficulty
in judgment, together with imperfections, perhaps inevitable, in
research data which lead regulatory agencies to give undue emphasis
in this situation to physical water quality standards or policies.
6.	We believe that water quality standards and bays and estuaries
policies are essential policy statements to provide guidelines to
industry and others regarding measures they would have to adopt to
operate their business. In most cases, these would apply. How-
ever, we believe that there are certain circumstances where
physical water quality standards as defined by the regulatory
agency are in direct conflict with ecologically more attractive
alternatives. In our judgment bioenhancement from biodegradable
non-toxic wastes such as tuna cannery effluent is an example of
such a case. Finding for bioenhancement should not be precluded
by pre-existence of a Water Quality Standard or a Bays and
Estuaries Policy. Unless these standards or policies are made
sufficiently liberal to cover all cases (which we do not
advocate), then if variances are not permitted, they will force
decisions to be made inequitably in the particular case where
bioenhancement is a factor.
7.	Although in theory a mechanism exists for permitting discharges
on the grounds of bioenhancement, we would suggest that the
burden of proof is being used unreasonably to prevent such
permits becoming a reality.
We should, therefore, like to see policy guidelines drawn up for regulatory bodies
and industry which would indicate the manner in which the question of bioenhance-
ment for biodegradable non-toxic wastes should be addressed. We should like to
see these guidelines indicate how judgment should be applied rather than
indicating what specific parameters and criteria have to be met. We believe these
guidelines should address the need for flexibility in interpreting water quality,
standards and other general policies and issuing variances where it is found on
balance to be the appropriate solution.
Yours truly
STAB-KIST FOODS, INC
Dave Ballands
General Manager, Engineering Services
-------
September 14, 1979
Mr. Calvin J. Dysinger
Project Officer for the Food
Industries Branch
Environmental Protection Agency
401 M Street, S. W.
Washington, D. C. 20460
Dear Cal:
We welcome this opportunity to comment on Dr. Soule's
Report, "Ecological Changes in Outer Los Angeles-Long Beach
Harbors Following Initiation of Secondary Waste Treatment and
Cessation of Fish Cannery Waste Effluent."
We have carefully monitored Dr. Soule's work throughout
its entirety and we have analyzed the Report dated April 1979.
As you know, the work so far is unquestionably an indication
of bioenhancement. It falls short only from a positive
demonstration of bioenhancement because fish canners in the
Los Angeles Harbor were required to hook up to the municipal
treatment facility, thus terminating the rich levels of fish
cannery waste effluent that formed the basis of this important
study. The preliminary results of the study Report, however,
present sufficient evidence to justify the continuation of
managed discharge of cannery waste in order to carry on with
the study for an extended period of time. However difficult,
we believe the continuation of the study will immeasurably
contribute to a definition of bioenhancement.
Since the issuance of the Report in April, we have
received numerous commentaries on the study and its conclu-
sions. I have enclosed for your information, copies of several
letters that may be of interest. In general, we believe there
is broad national support among scientific groups, environ-
mentalists, industry, and local governments for guidelines and
policies in dealing with bioenhancement properties of biode-
gradable non-toxic seafood cannery wastes.
r^3
TUNA RESEARCH FOUNDATION, INC.
SUITE 603 • 1101 SEVENTEENTH STREET, N.W • WASHINGTON, D. C. 20036 • (202) 296-4630
-------
Mr. Calvin J. Dysinger
September 14, 1979
Page 2
We urge that EPA accord to Dr. Soule's work the highesi
level of attention, and perspective as you prepare for the
Seafood Study Report to Congress as mandated under Section 74
of the Clean Water Act.
Respectfully yours
John P. Mulligan
President
Enclosures
JPM:jj
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(§nlf Jliates JSHarme ^ishtvhs (Eommissitm
MEMBER STATES
ALABAMA
FLORIDA
LOUISIANA
MISSISSIPPI
TEXAS
;%"V
/U 'S' S ^
-.*«• *.r
July 2, 1979
onoo
P.O. BOX 726
OCEAN SPRINGS, MS.
39564
(601)875-5912
JUL r

John P. Mulligan
Tuna Research Foundation, Inc.
1101 Seventeenth Street, N.W.
Washington, D. C. 20039
1379
fO^AT
Dear Mr. Mulligan:
I have just finished reading Dr. Soule's study,
"Ecological Changes in Outer Los Angeles-Long Beach
Harbors Following Initiation of Secondary Waste
Treatment and Cessation of Fish Cannery Waste Effluent."
I must say the scientific methodology seemed to reflect
an accurate and complete picture of the ecological
ij	situation in the harbors.
i-l i •
We feel this study brings into focus the real effects
that some agencies' mandated regulations actually have
ir.	on the environment. That is, the net effect of the
fj \	regulation did not enhance the situation, but led to
further complications which were not in the best interest
i|.' y of the environment. Now don't misunderstand, we have a
fji!< keen interest and respect for the environment, and do not
'!¦ purport to sacrifice any part for the sake of convenience
; | ' J: or economics. However, in the light of the findings in
I ; ' the study, there is an opportunity to enhance the marine
•	area with the rich nutrients that do not have a use in
i	the cannery.
We feel the return of the biological wastes to the environ-
ment is much different than the release of metals, and
i£;, /' toxic chemicals, which do not add to the nutrient value
-------
Page 2
of the ecosystem. With the realization that the ecosystem
cannot cope with unlimited Telease of biological wastes,
we agree that managed levels of cannery wastes can and
should be placed back into the ecosystem.
We feel possibly this study might prompt investigation
on the effects of released fertility on the enhancement
of mendaden stocks of the Gulf of Mexico. In any event
the study did address some questions many people have
had about the benefits waste effluents have on the over-
all ecosystem.
Thank you for the opportunity to comment on this study.
Larry 3. Simpson
Sincerely,
Assistant to the Director
LBS/ca
-------
UNIVERSITY OF DELAWARE
COLLEGE OF MARINE STUDIES
LEWES COMPLEX
PHONE: 302-6*5- 4274
LEWES. DELAWARE
1 9958
AUG 6 1979 ^
August 2, 1979
HJNA RESEARCH FOUNDATION
Mr. John P. Mulligan, President
Tuna Research Foundation, Inc.
Suite 607
1101 Seventeenth Street, NW
Washington, DC 20036
Dear Mr. Mulligan:
I examined with interest a copy of the recently completed study con-
ducted at USC's Institute for Marine and Coastal Studies, Allan Hancock
Foundation, entitled "Ecological changes in outer Los Angeles-Long Beach
Harbors following initiation of secondary waste treatment and cessation
of fish cannery waste effluent."
Dr. Dorothy F. Soule and her associates are to be complimented for
conducting such a thorough and timely study. The conversion of the harbor
from a rich soft-bottom community with a rich biota to a less productive
one coincident with diversion of the fish cannery effluents from the harbor
into the treatment plant, is clearly demonstrated. I find it difficult to
disagree with the report that release of managed levels of cannery wastes
into the harbor without secondary treatment of those wastes would create a
better nutrient balance in conjunction with secondary TITP wastes, and would
be beneficial to the biotic life in the area.
I find the report especially interesting, because the richest oyster
growing estuary, by far, in our area, is Broadkill River which receives con-
siderable (it has never been carefully measured) organic wastes from a clam
processing plant a short distance up the estuary from the spot where we
locate our floats for experimental purposes.
The report deserves the serious consideration of EPA personnel in the
completion of their Seafood Study report to Congress as mandated in Section
74 of the Clean Water Act of 1977.
Yours sincerely
/
Mel bourne RCarri ker
/
t.
MRC/dp
cc: Dr. Dorothy F. Soule
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PURDUE
UNIVERSITY,

//
FOOD SCIENCES INSTITUTE
September 10, 1979
Ms. Helen K. Brock
Director of Government Relations
TUNA Research Foundation, Inc.
1101 Seventeenth Street, N.W.
Suite 603
SEP 1 3 J979 ^
MA RESEMf FOUMATIOH
Washington, DC 20036
Dear Ms. Brock:
Thank you for providing me with a copy of the Soule and
Oguri study dealing with the ecological changes in the Los' •
Angeles - Long Beach Harbor areas. I have reviewed the summary
report provided me and found this report interesting. How-!
ever, their findings were not unpredictable since microflora •
and fauna populations are unquestionably dictated by the.food
and food chain relationships that exist within a given defined
region. The Soule-Oguri simply documents that relationship for •
the Los Angeles - Long Beach Harbor areas and correlates these .
population shifts with "bioenhancement" derived from the fish
cannery discharge.
As I assess the TUNA Research Foundation's involvement, I
see the Foundation's position as supporting regulations to
allow for direct discharge to the Harbor areas by the fish-.',
canneries. The benefits obviously would be lower operating-
costs (DAF unit and solids waste disposal) for the fish canneries
and more fish would be potentially caught by the commercial. .
fishermen.
A key to this position is how PL 92-500, Title I, Section t
101 (a), (1) (2), and (5) are interpreted by EPA, state*and
city regulators. An argument for the TUNA Research Foundation's'
position can be found in the defined goals of the Federal
Water Pollution Control Act of 1972. Title I, Section 101 (a)
of this act states "The objective of the Act is to restore.and
maintain the chemical, physical and biological integrity .pf
the nation's waters." Underlined words are relevant to Founda-
tion's position. Also important to the argument is item'(2)
of the objectives which states "it is the national goal that
whenever attainable, an interim goal of water quality which". .
Q Smith Hall
JO West Lafayette, Indiana 47907
-------
2
provides for the protection and propagation of fish, shellfish,
and wildlife (e.g. birds) and provides for recreation in and on
the water be achieved by July 1, 1983;". What needs to be defined
for the Los Angeles and Long Beach Harbor areas is the desired
"water quality". Maybe "quality" as applied to fresh water does
not have the same meaning as for salt water areas.
For the most part, EPA wishes to eliminate the discharge
of pollutants from navigable waters and develop areawide waste
treatment management and planning processes to assure adequate
control of point source pollutants in each state. To this end,
the EPA has required pretreatment of fish cannery wastewater
and subsequent treatment by a municipal waste treatment process
before discharged to the harbor waters. At issue is the esthetics
of the harbor areas and where the priorities are to be on the
environmental quality for these areas (commercial vs recreational
and residential). Frankly, the answer to this issue is in the
local (Los Angeles) political arena. If support can be gained
there, the chances for allowing for fish cannery wastewaters to
be discharged to the harbor will be enhanced.
If I can be of further service to you, please do not hesitate
to- contact me accordingly.
Sincerely yours,
James V. Chambers, PhD
Associate Professor of Animal
Sciences and Extension Specialist
Phone: (317) 494-7825
JVC/jl
P.S. I was an instructor in the workshop for extension specialists.
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GULF OF MEXICO FISHERY MANAGEMENT COUNCIL]
	Lincoln Center, Suite 881 • 5401 W. Kennedy Blvd. \
Tampa, Florida 33609 • Phone: 813/228-2815
June 27, 1979
Mr. John P. Mulligan, President	°ul- «>|j,3
Tuna Research
1101 17th Street, N.tf.	IUNA KloEAHGH F0Ui';0AT»0N
Suite 607
Washington, D.C. 20036
Dear Mr. Mulligan:
We have reviewed with interest the report sent to the Gulf Council
related to effluent discharges from tuna canneries in the Los Angeles
Harbor area. From the report's findings, it would seem that strict
and narrow adherence to the Clean Water Act actually resulted; in
this case, to defeat the spirit of the Act.
Your stated aim to include this report in the EPA Seafood Study
meets with our approval. This report points out the need for
examining the appropriate type of discharge which should be per-
mitted for the surrounding environment. We believe it also points
out the need for similar studies for different types of seafood
processing, where bioenhancement goals or properties of the dis-
charge may be different.
Thank you for the opportunity to review this material.
Sincerely,
A council authorized by Public Law 94-265, the Fishery Conservation & Management Act of 1976
Wayne t. Swingle
Executive Director
WES:VB:mjl
I

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October 19, 1979
Mr. Calvin J. Dysinger
Project Officer for the Food
Industries Branch
Environmental Protection Agency
401 M Street, S. W.
Washington, D. C. 20460
Dear Mr. Dysinger:
This is in further reference to Mr. Mulligan's letter
of September 14, 1979 regarding Dr. Soule's Report, "Ecological
Changes in Outer Los Angeles-Long Beach Harbors Following
Initiation of Secondary Waste Treatment and Cessation of Fish
Cannery Waste Effluent."
Enclosed is a copy of a letter from June Lindstedt Siva,
Senior Science Advisor, Environmental Sciences, Atlantic Richfield
Company, commenting on the report.
I hope this information will be helpful to you in your
review of documents related to the ecology of the Los Angeles-
Long Beach Harbor.
TUNA RESEARCH FOUNDATION, INC.
SUITE 603 • 1101 SEVENTEENTH STREET, N.W. • WASHINGTON, D. C. 20036 • (202) 296-4630
Sincerely
Helen K. Brock
Director of Government Relations
Enclosure
-------
515 South Flower Street
Mailing Address: Box 2679 * T.A.
Los Angeles, California 90051
Telephone 213 486 0741
June Llndstedt Siva, Ph.D.
Senior Science Advisor
Environmental Sciences
October 3, 1979
Dr. Dorothy Soule and
Mr. Mickey Oguri
Allan Hancock Foundation
University of Southern California
Los Angeles, CA 90007
Dear Dorothy and Mickey:
You are to be congratulated for putting together such
a comprehensive report on the ecology of Los Angeles-
Long Beach Harbor. This is one area where it now might
be said that there is a "baseline." I am much impressed
with the participation in the study of a good part of
the local scientific community. The report is not only
a significant contribution to the scientific literature,
but also can be used as a valuable planning tool when
new projects are proposed for the harbor area. I wish
we had similar studies for other coastal areas. They
would certainly make it easier for us to make ecolog-
ically sound coastal zone management decisions.
Again, congratulations on an excellent report.
Sincerely,
June Lindstedt Siva
JLS/lr
/u£ ftssre t/tS/C rr
-------
National Food Processors Ansociation
1133 Twentieth Street N.W., Washington, D C. 20036
Telephone 202/321 - 5900
Agricultural and
Environmental Affaire
Edwin A. Crosby, Ph.D.
Senior Vice President
202/331-5967
Jack L. Cooper
Director,
Environmental Affairs
202/331-5968
Raymond F. Aitevogt, Ph.D.
Assistant Director,
Agricultural Affairs
202/331-5969
September 14, 1979
Mr. Calvin J. Dysinger
Effluent Guidelines Division (WH-552)
U. S. Environmental Protection Agency
401 M Street, S. W.
Washington, D. C. 20460
Regarding: NFPA comments on the report Ecological Changes in Outer Los
Angeles-Long Beach Harbors following Initiation of Secondary
Waste Treatment and Cessation of Fish Cannery Wastes Effluent,
a report for the city of Los Angeles Department of Public Works,
Bureau of Engineering, for the Terminal Island Treatment Plant
and the Environmental Protection Agency, Report to Congress on
Seafood Waste Effluent, for the Tuna Research Foundation by
Dorothy F. Soule and Mikihiko Oguri of the Harbors Environmental
Projects, University of Southern California, Los Angeles, California,
Marine Studies of San Pedro Bay, California, Part 16, April 1979
NFPA staff and several members of our Effluent Guidelines Subcommittee
for Seafoods have reviewed the above report. We conclude that a good case has
been made for bioenhancement from the study of discharges from tuna canneries
at Terminal Island. The seafood processing industry believes that bioenhancement
is a valid concept and urges the Agency to include a thorough discussion of it,
including a proposed definition of bioenhancement, in its Report to Congress
required by Section 74 of the Clean Water Act.
Dear Cal:
Sincerely

Jack L. Cooper
cc: Effluent Guidelines Subcommittee for Seafoods
3
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-2 1
EXCERPT FROM UNPU3LISHED MANUSCRIPT
Comparison of KEP and Edison Data
Souie and Oguri (1979) showed the decrease in the mean
number of fish per trawl from the 1971-73 oeriod through 1978
from 423.2 fish per trawl (or over 700 if larval fish are
included) to under 100 fish per trawl for the period of Decem-
ber 1977-October 1978.
While the Southern California Edison Report (EQA-MBC, 1978
stated that Long Beach Harbor supports a large, healthy, stable
and diverse population of fishes that are common to nearshore
southern California waters, and indicated that there was a
large increase in mean harbor fish populations, analysis of the
Edison data and HEP data shows agreement in major trends as
well as the differences in species composition.
Trawl Data and Trends
The HEP trawl data taken by Dr. J. S. SteD'nens for the
outer harbor were gathered in daylight hours. The data were
plotted (Soule and Oguri, 1979) both as annual means, between
1973 and 1978, and as seasonal means. Dr. Stephens calculated
that the HEP mean number of fish for 1974-197 6 was 212 per
trawl, whereas he calculated the Edison mean for that same
period as 180 fish per trawl, a fairly close agreement for the
two different areas. However, the Edison data were summed and
thus no trends could be shown; a four-way analysis by Stephens
indicated significant annual variation but that cannot indicate
direction.
There were several major changes in the harbor between 197
andl978. In 1975 Dissolved Air Flotation ("primary treatment")
was installed on cannery waste streams. In January 1977, the
operational phase of the Long Beach Edison plant began, while
in April 1977 secondary treatment was initiated at the Terminal
Island Treatment Plant in outer Los Angeles Harbor. The first
cannery outfall was diverted to TITP in October 1977 and the
second in January 1978, closing off those effluents from enter-
ing the harbor except after secondary treatment.
Stephens (in Soule and Oguri, 1979) mentioned the defici-
ency of outer harbor trawl data during 1975-1977. The Edison
trawls in 1977 far outnumbered the casual trawls carried out
by Stephens and his Occidental College students, who provided
their data for the Soule and Oguri report. However-, HEP trawls
numbered 55 between December 1977 and October 1978 for the
Terminal Island Treatment Plant Study. Edison trawl data are
available only through March 1978. in Figure 3 the mean trawl
data for the Edison plant are plotted against the HEP mean
data for 1971-1976. Extreme variation can be seen in the
^
-------
Edison trawl numbers in 1977, almost a "yo-yo" effect to the
lines; seasonal means have been combined to give comparable
data points for both Edison and HEP data. In Figure 4, all
Edison trawl data for 1977 are plotted, along with a separate
plot for the mean data from the two Edison trawl stations
(T13 and T15) nearest the HEP stations, and for HEP 1977 data
(data from EQA-M3C, 1978; Soule and Oguri, 1979). The 1978
HEP data continues through October 1978. The lower plot (based
on data in EQA-MBC, 1978) shows the variations in the Edison
plant operation, which was reported as percent generating
capacity, during the 1977-1978 period.
When the Edison mean data are broken into the same time
periods as the HEP data, the trends are strikingly similar
(Figure 5). The mean data from all Edison daytime trawls (AED)
were plotted; then the means for two Edison trawl stations T13
and T15 in the outer harbor (OED) closest to HEP stations were
plotted separately, and the HEP means were also plotted. These
data were all recalculated into the same time periods reported
in Soule and Oguri (1979) for greater comparability; the
periods were January-June 1977, July-November 1977, December
1977/January 197S and March/April 1978. Edison trawls were
not reported after March 1978, but April, July and October
1978 mean trawls for HEP were plotted.
The Edison trawl data showed much higher 1977 means; 508
fish for all stations (daytime) and 336 for T13 and T15 in the
outer harbor in the Long Beach Pier J-Channel area, as com-
pared with 216 in the HEP total outer harbor area. However,,
for the Edison trawls, the trends in the means from January-
June levels to the July-November 1977 levels were strongly
down. The means were further depressed in December 1977/January
1978, with an outer harbor Edison (OED) mean of 24.5 fish and
an HEP mean of 26.7 fish. These are, as Stephens indicated
(Soule and Oguri, 1979), unprecedentealy low trawl numbers.
The mean of 76.5 for all Edison trawls for January 197 8 is
hardly much better. The smallest symbol used on Stephens'
et at. (1974) Figure 4 on mean abundance was 85-90 fish. In
Soule and Oguri (1979), Figure 8 shows large areas with fewer
than 10 fish per trawl in December 1977, with only the outfalls
area having 155 fish; that station provided the only numbers
of consequence to give the mean cf 26.7 fish to the period.
It is normal for fish counts to be low in winter, but this
seemed impoverished. It also coincided in time with removal of
fish cannery waste effluents from the outer harbor, reoresent-
ing an enormous drop in available energy (calories) to the eco-
system. The December 1977 trawls came before the major rainfall
at the end of the month, and the water was still warm. Fish
will sometimes leave the harbor as a storm front approaches
1 it this did not aDpear to be the case then.
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-600
	O	 HEP x trawl data
!	1 HEP x for ¦period
AED x alt Ed. stations
i	i AED x for period
	o	CED x outer harbor Ed.
I	1 QED x for period
-500
SOS
\
x AED
500
\
3ZS \ , x CED
x OED
x HEP 94.6
Jl-Nov
77
Dec 77/
Jan 73
Jul 78
Oct 78
Figure 5. Comparison of HEP Outer Harbor Data and EQA-MSC Edi-
son Data Adjusted to Season and Daytime for All Stations (AED)
and for Outer Harbor Edison Trawls (T13, T15) Only (OED).
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Summary of Issues - L.A. Harbor Enhancement Study
P. T. Brubaker
Y^/77
Files
1. Historical Background
Discharges from four Terminal Island fish canneries (Van
Camp Seafood Company# Pan Pacific Fisheries and Star-Kist
Poods Plants 1 and 4) have long been associated with water
quality degradation in Los Angeles Harbor. Current efforts
to solve this problem may be traced back to the early 1970's,
when State and Federal actions culminated in 1) the funding
of planning and construction for upgrading the City of Los
Angeles Terminal Island Treatment Plant (TITP), to include
adequate capacity and treatment facilities to provide
secondary treatment to domestic and fish cannery wastes,
and 2) written commitments from the fish canneries to EPA
to install primary treatment facilities and to direct their
wastewaters to TITP v;hen the upgrading was completed. In
1971, the Harbors Environmental Projects of the the Allan
Hancock Foundation, affiliated with the University of
Southern California, commenced a study of Los Angeles Harbor.
In 1973, HPDES permits were issued to the canneries by the
State of California, requiring installation of primary
treatment facilities.
In May 1974, the State of California adopted the "Kater
Quality Control Policy for the Enclosed Bays and Estuaries
of California" (Bays and Estuaries Policy, or BEP). This
policy was approved by EPA as a State/Federal Kater Quality
Standard, pursuant to Section 303, FWPCA, on January 8,
1976. The BEP provides that all municipal and process
wastewater discharges "shall be phased out at the earliest
practicable date," unless it can be shov/n that a discharge
"enhances" the quality of the receiving water and is
non-toxic.
Through 1974, the canneries proceeded to install primary
treatment facilities (disolved air floatation units, defined
by EPA in 1974 as Eest Practicable Control Technology Cur-
rently Available, or, BPCTA). Construction of TITP upgraded
Reprod
best av
duced from
vailable copy.
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facilities, originally planned for 1974, was delayed; incre-
mental delays pushed the completion date back to r.id-1977.
Rather than directly addressing compliance with water quality
standards, the expiration dates of the cannery HPDES permits
vere revised to coincide with the the expected TITP completion
date. Also, in 1974, the canneries' effluent limits were
modified to be consistent with promulgated guidelines for
BPCTCA. In December 1976, the Enhancement Study document
was completed and submitted to EPA and the State by the
canneries.
On February 9, 1977, EPA informed the State that new permits
for the canneries must require compliance with approved
water quality standards (the BEP), and that no discharge of
process wastes could be allowed absent a Finding of Enhance-
ment. Concurrently, EPA conducted a review of the enhancement
study document. EPA's reviews of the document established
that it does not show enhancement, and that, in fact, it
includes considerable evidence that the cannery discharges
are detrimental to the receiving waters, and do not meet
minimum toxicity criteria. In June 1977, the State issued
new NPDES Permits to the canneries which required compliance
with water quality standards by July 1, 1977# i.e., the
elimination of process waste discharges.
On June 8, 1977, EPA brought suit against two of the canneries
for repeated violations of effluent limits. On August 26,
1977, actions were filed against the remaining two canneries.
The object of the suits was to compel the canneries to com-
ply with the water quality standards, and to pay penalties
for past non-compliance with the permits.
2. Enhancement Study Issues
The study document represents a portion of a long term
continuing study of the harbor area by the Hancock Founda-
tion. The study is identified as "Marine Studies of San
Pedro, California, Part 12, December 1976." It is supported
by funds from sources including the City of Los Angeles, the
tuna industry and the U.S. Office of Sea Grant Programs.
The study document is a collection of research papers covering
a variety of physical and biological areas of interest in
L.A. Harbor.
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\
-3-
The basic premises of the enhancement argument are that
the cannery process wastes (characterized as high in BOD,
proteinaceous suspended solids and oil & grease) provide
nutrients necessary to the sustenance of a largo fish popu-
lation in the harbor, and that this benefit overrides the
facts that the cannery wastes show toxic effects up to one-
half mile from the cannery outfalls and that the wastes
create a marked depletion of dissolved oxygen over a large
part of the harbor.
EPA reviews of the study conclude that the research reports
do not support the enhancement contention.
First, the study does not establish that the existing,
•enhanced," conditions represent a iaore desirable state than
existed prior to harbor development or than exists in any
comparable control area. There is no evidence that the
existing situation is in fact the optimum for the harbor.
It is indicated that the overall health of the harbor eco-
system has shown improvement since the institution of improved
wastewater treatment by the canneries in 1974-5.
Second, the study does not show that the existence of any
fish population is dependent upon the cannery discharges for
survival; in other words, the discharges do not provide
relief from a nutrient-limited conditions. The negative
effects of the discharges on the ecology of the harbor, such
as demonstrated acute toxicity and depression of dissolved
oxygen levels, axe not adequately explored.
Third, the study is compromised by numerous methodological
shortcomings and omissions, particularly in the area of
determining the contribution of factors other than waste-
water discharges to the existing and projected conditions.
Fourth, in the context*-of the BEP, which requires compliance
with specific toxicity standards as one condition of an
enhancement finding, the report documents numerous toxic
effects of the cannery wastes on a variety of organisms.
These data alone would disqualify the cannery discharges
from consideration for an enhancement finding. These toxic
effects are net limited to undiluted influent, they are also
apparent over a large portion, possibly as large as several
hundred acres, of the harbor.
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4-
In summary, EPA finds that the study does not demonstrate
enhancement, nor does it provide evidence that the cannery
discharges can meet, rainiEiura toxicity standards. Moreover,
given the evidence at hand, EPA does not believe that there
is a reasonable chance these conclusions will be altered if
the study of existing conditions were to continue. EPA has
no objections to the continuation of the study after the
elimination of the discharges to the harbor.
Brubaker/RJGleason
Reading File
362661 et al
8/30/77
9/26/77:RJG
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