-------
1974 - 23 SPECIES
//77/S//////S////; CHRYSOPHYTA
CHLOROPHYTA
1975 - 42 SPECIES
V////////.//////\ EUGLENOPHYTA
////W7/S//V fcHRYSOPHYTA
CHLOROPHYTA
1976 - 48 SPECIES
//////////T/77. EUGLENOPHYTA
///////////A
.CYANOPHYTA
V7//777//77S/7/////////. CHLOROPHYTA
TOTAL 1974-1976 - 5O SPECIES
EUGLENOPHYTA
CYANOPHYTA
Y77//////////.//////////A CHLOROPHYTA
10 .
20 25 30 35 4O 45 50
COMPOSITION
Figure 20. Percent species composition, by division,
for Lake
OF SPECIES (%}
B over a three-year period.
46
-------
Algal standing crops in Lake B (Figure 2 !)••-fluctuated
dramatically over a two-year period, The 'Cyanqphyta reached
maximum peaks during the summer, while:"maximum ^standing .crops
of Chrysophyta occurred in winter months,, The' green algae
obtained maximum cell numbers in September, but .euglenoids
dominated the algal standing crop during this month. Algal
numbers in Lake B increased during the-second year.>o,f study due
primarily to larger standing crops of blue-green;algae. Subse-
quent studies indicated no significant increases in phytoplankton
from 'November, 1975 to May, 1976. However, a dense phytoplankton
community composed of Oedogonium and-Spirogyra developed in the
littoral zone of this lake.
Since algal species differ in size, cell numbers, do-not
always reflect actual biomass. Therefore, total,cell numbers.
were converted to dry weight-and cell -.volume estimates by use
of tables available in the literature (24,26,28) and calculations
performed in our laboratory. Figure 2^2 . presents ,,estimates of
algal biomass by division on the basis of percent' of total, dry
weight. Euglenoids dominated algal biomass, except in February
of 1975 when diatoms were dominant. Chrysophycean algae were
major codominants during winter months^ while cyanophycean algae
were major codominants in summer months. .Chlorophycean algae
were minor contributors to algal solids in the lake.
Algal populations in Lake B were relatively low when com-
pared to other aquatic ecosysterns^ For example/ cell numbers
exceeding 104 cells/ml have been reported for Lake Michigan (29)
and in highly nutrified water in southeastern: Texas (30) . Algal
cell numbers and biomass in Lake B .were .well,-below the 106 cells/
1 (31) or 8 mg/1 (32) values reported as being .sufficient to
classify algal growth as a water bloom. Algal 'cell volume in
Lake B was also well below the bloom proportion reported by
Kramer et al. (31) . Average total aig'al -biomass ,:ihf Lake B was
38.88 yg/1, while average suspended solids was 603 mg/1 '(Table
14) . Thus, algae were a minor .part of the particulate material
in Lake B, and suspended solids were definitely not' of algal
origin. ' •••'.;' ...'.'•
According to Vollenweider (33) , -'algae'are composed of 40-
60% carbon by weight. On the basis' of V-''5rb'% carbQn content,
total dry weight values per month: were converted .to carbon
estimates (Table 14) . These conversions .indicated- that 'algae
contributed little to organic .loads of ' Lake-B,; since, the average
concentration of total organic carbon was 16.;8.5 mg/1. 'The
average rate of change in algal carbon,.based on-gains and
losses in algal biomass at 30-day intervals,,was 0.14 mg C/m2/
day. Even though this figure is not-.-a -daily rate of carbon
production, it can be used to make general comparisons between
algal carbon in Lake B and other aquatic .ecosystems. For
example, Rodhe (34) reported algal production values .of 7-25 g
C/m2/yr for oligotrophic lakes, 75-250 g*C/m.2/yr for naturally
47 - - ,'. ; ' " '
-------
1.0 .
0.9 .
O.S.
0.7
3
UJ 0.5
o
IO
c
0.4
0.3
0.2
O.I
2.9
• • CHLOROPHYTA
0 0 CYANOPHYTA
x K CHRYSOPHYTA
A A EUGLENOPHYTA
O « TOTAL
QJFMAMJJAiSONDJFMAMJJASON
I MONTHS
Figure 21.
Seasonal algal standing crops
in Lake B.
48
-------
PQ
o
0
o
0>
0
a>
o
h-
o
(T>
o
vn
o
^r
0
to
o
CNJ
o
o
d
tn
H
(%) 1H9I3M AUd
49
-------
TABLE 14. COMPARISON OF ALGAL BIOMASS AND CARBON CONTENT
CONTRIBUTED BYiEACH ALGAL DIVISION
1
Algal
Division
Chlorophyta
Cyanophyta
Chrysophyta
Euglenophyta
Total
I
Weight Range . Average Weight
(ug/D (wg/D
0.24 -
0.52 -
0.42 -
6.5 -
11.6 -
2.16 :
6.50
18.48
377.00
380.10
0.74
2.80
4.62
30.63
38.88
Average Carbon
Content (yg/1)
0.37
1.40
2.31
15.32
19.44
eutrophic lakes, and 350-700gC/m /yr for culturally eutrophic
lakes. Even though these data are subject to interpretation,
they provide a general picture of primary productivity in lakes
with various trophic classifications. More specifically, Wetzel
(24) reported annual productivity values (g C/m /yr) of 36 for
Castle Lake, California (oligotrophic), 160 for Clear Lake,
California (mesotrophic), and 369 for Lake Wintergreen, Michigan
(eutrophic). Lake B probably had productivity rates resembling
those of oligotrophic waters 1. Even considering loss of algal
calls by grazing or sedimentation, it is unlikely that Lake B
had productivity rates approaching those of mesotrophic or
eutrophic lakes.
Multiple regression indicated that the measured physico-
chemical parameters in Table 15 influenced algal numbers in Lake
B. The relatively low regression values indicate that measured
physicochemical parameters were not responsible for total changes
in algal numbers in this lake. Other factors or interactions of
various factors may have exerted more impact on changes. The
composition and standing crops of algae were probably signifi-
cantly influenced by the recirculation of water in the lake and
by the use of Panther Branch water to maintain the constant
volume lake.
I
CONFERENCE CENTER LAKES—NUTRIENT LIMITATIONS
Nutrient limitation studies with low-flow water collected
from Lake B revealed that algal growth was not stimulated by
addition of nitrogen to the water (Figures 23 and 24). However,
phosphorus spikes did stimulate algal growth, thus indicating
that phosphorus was the limiting nutrient for algal growth in
Lake B and Lake A (Figure 24). The relatively high levels of
growth with combined spikes also indicated that algal numbers
could be increased substantially by additions of both nitrogen
and phosphorus to the water and that other nutrients were not
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53
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particularly limiting for algal growth. However, bioassays with
stormwater runoff collected from Lakes A and B (Figure 25) in-
dicated that nitrogen was mo^re limiting than phosphorus for
algal growth. j
The data of Miller et al. (27), previously discussed, were
used to calculate theoretical algal yields from measured concen-
trations of TSIN and PO4-P ih Lake B (Table 16). These data in-
dicate that theoretical yiel&s were substantially higher than
observed algal biomass provided that some factor, other than
nitrogen or phosphorus, was not limiting for algal growth.
Ratios of nitrogen and phosphorus may be used to estimate which
of these nutrients is potentially limiting for production of
algal biomass. Miller et all (27) and Chiaudani and Vighi (35)
reported optimum N:P ratios 6f 11.3 and 10, respectively, for
growth of Selenastrum capricprnutum. A ratio of 15:1 was re-
ported as optimum for algal growth by Fitzgerald (36), while
Vollenweider (33) reported a; range of 10-15 as optimum. On the
basis of a 10:1 ratio, phosphorus is a potential limiting nutri-
ent for algal growth during some months, while nitrogen is
potentially limiting in other months (Table 16). However, algal
biomass in Lake B never exceeded theoretical yields; thus, these
nutrients were probably not the primary limiting factor in the
lake.
Bioassays indicated that low-flow water from Lake B could
support a biomass of S_. capricornuturn which exceeded recorded
algal biomass. A filtered and autoclaved sample produced 27 mg/1
dry weight of £. capricornuturn, while a portion of the same
sample autoclaved before filtration suppored a yield of 50 mg/1.
Autoclaving of lake water seamed to solubilize nutrients which
could be utilized for growth;by the test alga. Bioassays with
unautoclaved, filter sterilized water probably provided low
estimates of growth supporting capabilities of Lake B water.
Yields of S^. capr icornu turn were also increased by removal of
turbidity (suspended solids)'from Lake B water by use of various
size filters. A 97% reduction in turbidity resulted in a 94%
increase in algal yield, while a 56% turbidity reduction
increased algal yield by 84%. Nutrient spikes also indicated
that phosphorus additions stimulated algal growth in the majority
of Lake B water samples. However, nitrogen was, on occasion, the
limiting nutrient for algal growth in low-flow water and was
always the limiting nutrientjin surface runoff collected during
storm events. Correlations between theoretical yields, observed
biomass, cell volume, and bioassay yields are not possible since
water samples for chemical analyses and algal bioassays were not
collected simultaneously. The stimulation of algal growth by
nitrogen and phosphorus spikes does indicate that other nutri-
ents were present in quantities sufficient to support added
growth of the test alga. Thus, nitrogen and phosphorus were
primary limiting nutrients, but were not necessarily the limiting
growth factors in Lake B. j
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TABLE 16. THEORETICAL YIELDS OP ALGAE AND N:P RATIOS BASED ON
NITROGEN AND PHOSPHORUS CONCENTRATIONS OF LAKE B
I
1
Month
Dec.
Jan.
Feb.
Mar.
Jun.
Jul.
Aug.
Sept.
Oct.
Jan.
Feb.
Jul.
Aug.
Sept.
Oct.
TSIN+
(mg/1)
0.18
0.33
0.22
0.17
0.21
0.22
0.18
0.35
0.14
0.20
0.10
0.06
0.11
0.74
0.34
P04-P
(mg/1)
0.08
0.01
0.05
0.07
0.05
0.01
0.10
0.06
0.05
0.01
0.01
0.09
0.03
0.05
0.04
1
1
I
1
N:P
i
2.3:1
33:1
4.^:1
2.jl:l
4.h:l
1
22^1
i
1.8:1
i
5.^:1
I
2.0:1
1
20?1
lOpl
1
1:1.5
i
3.^:1
141.8:1
8.^:1
\
\
TSIN
Yield
(rag/1)
6.8
12.5
8.4
6.5
8.0
8.4
6.8
13.3
5.3
7.6
3.8
'2.3
4.2
28.1
12.9
P04-P
Yield
(mg/1)
34.4
4.3
21.5
30.1
21.5
4.3
' 43.0
25.8
21.5
4.3
4.3
38.7
12.9
21.5
17.2
Recorded
Algal
Weight
(yg/D
0.022
0.015
0.012
0.013
0.016
0.015
0.036
0.380
0.014
0.044
0.031
0.017
0.028
0.033
0.052
1
1
Theoretical yields based oni conversion factors of Miller
et al. (27)
TSIN: total soluble inorganic nitrogen
(NH3-N + N03-N + N02-N)
56
-------
Lake B parameters were compared to those of lakes with
various trophic levels (24). Total organic carbon and phos-
phorus concentrations in Lake B resemble those of eutrophic
lakes, while total nitrogen falls within the range of oligo-
mesotrophic lakes. However, phytoplankton biomass and algal
carbon indicated that Lake B was ultra-oligotrophic. Nutrient
supplies seemed sufficient to support algal standing crops and
biomass in excess of those actually observed in Lake B.
The high initial turbidity of Lake B probably limited algal
diversity to those species physiologically or morphologically
adapted for existence at low-light intensities. Reductions in
turbidity seemed to allow a more diverse assemblage of algae,
composed primarily of chlorophycean species, to develop in the
lake. However, even with reduced turbidity, algal biomass was
dominated by euglenoids and chrysophytes or blue-green algae.
Growth of green algae may have been limited by the high pH, high
turbidity, or a combination of these two factors. The relatively
low algal biomass indicates that some factor, possibly turbidity,
was limiting algal growth. However, dense floating mats of
Spirogyra sp. and Oedogonium sp. developed in the littoral zones
of Lake B. Reduced turbidity would probably be accompanied by
comparatively higher yields of algae in Lake B, and, at this
time, algal growth would probably be limited by nitrogen and
phosphorus concentrations in the lake. Future reductions in lake
turbidity might necessitate some management strategy, such as
regulations of detention time or use of well water to dilute
nutrients entering the lake via treated sewage effluent or
Panther Branch water.
Since nitrate-nitrogen and orthophosphate-phosphorus contri-
bute heavily to algal blooms, it may be desirable to limit or
prohibit homeowner and golf course use of nitrat&^nitrogen and to
substitute ammonia or urea fertilizers for use in increasing lawn
fertility. Another source of both nitrogen and phosphorus could
be eliminated by reducing the amount of lawn clippings and tree
litter which could be transported to the lakes by urban runoff.
In urban areas, the first rains and runoff usually contain
significant quantities of dissolved solids, including trouble-
some amounts of nitrogen and phosphorus. Thus, it might be
advantageous from the standpoint of eutrophication to collect
the first stormwater runoff and treat it with the sewage. After
the first flush, stormwater could then be directed through the
lakes to achieve mixing and washout of accumulated bottom sedi-
ments.
During periods of low rainfall, when phosphorus would be
expected to accumulate in lakes and concentrations of phosphorus
approach those known to cause blooms under laboratory conditions,
it might become desirable to use phosphorus precipitating chemi-
cals to lower the phosphorus level below the critical value.
57
-------
Soil renovation of wastqwater is also practical under some
circumstances. It may be th&t the recreational lakes should be
designed so that groundwateriserves as the primary source of
recharge. Stormwater and sewage effluent could be percolated
through and renovated by a spreading basin before going to the
lakes as groundwater. <
The above possible strategies represent only a few that
could be utilized in maintaining a desired water quality in The
Woodlands lakes. These strategies may be neither necessary nor
financially justifiable unless there is high expectation that
one or more will help control eutrophication. The design of a
lake management program which will maintain the aesthetic and
recreational values of the lakes can only be achieved through a
detailed study of the biological and chemical components of that
habitat. \
EDAPHIC ALGAE |
As members of the terrestrial microbiota, algae may function
in the formation and stabilization of soils (37,38). Some blue-
green algae, notably species of Nostoe and Anabaena, contribute
to soil fertility through nitrogen fixation (39,40,41), and meta-
bolites excreted by algae may be utilized by other microorganisms
(38,42). Even though edaphic algae occupy such important posi-
tions in the ecology of terrestrial habitats, they have received
relatively little attention from phycologists. Relationships
between edaphic algae, soil pH, soil type, and macrovegetation
have been determined for various surface soils in the United
States (43,44,45,46,47,48,49)1. Few publications (41,50,51,52)
deal with standing crops of edaphic algae, and there is a paucity
of information oh the distribution of algae with depth in soils
of the United States '(52,53,54,55) . Previous work on the rela-
tionships between the,-distribution of soil algae and the chemis-
try of soils is unknown to the authors.
Thus, an investigation was conducted to determine the ef-
fects of various types of land usage on distribution of edaphic
algae. Qualitative and quantitative determinations of soil algal
populations, as related to soil chemistry and soil depth, were
made for several disturbed and undisturbed soils. The ability of
water leachates from various soils to support algal growth was
also evaluated.
i
Fifty-one sites were selected for collection of surface soil
samples (Figure 26). Twenty^five sites were located in disturbed
and 26 in undisturbed portions of the study area. Since some of
the sites were situated in and around the Conference Center
Lakes, a more detailed map of their distribution is presented in
Figure 27. Samples were taken from the lake basins before
inundation. A description of each site is presented in Table 17.
58
-------
5-1,5-2
3,5-4
Figure 26. Location of soil collection sites.
59
-------
S6
S9
SIO
0* 4OO* 800"
Figure 27. Location of soil collection sites
in Conference Center Lakes area.
60
-------
TABLE 17. DESCRIPTION OF SOIL COLLECTION SITES
Site No.
SI
S2
S3
S4
S5
86
S7
S3
S9
S10
Sll
S12
S13
S14
815
816
817
818
Al
A2
A3
Description Disturbed Fertilized
Woods - -
Lake Basin + -
Lake Basin + -
Lake Basin +
Woods
Woods
Woods
Woods - -
Lake Basin •¥ -
Woods - -
Woods - -
Lake Basin + -
Woods
Lake Basin •*• -
Woods - -
Lake Basin -f - .
Woods - -
Woods - -
Woods
Lake Shoreline +
Woods
PH
6.9
6.7
6.7
6.9
6.3
6.2
6.0
5.8
6.7
5.9
5.6
7.1
5.9
6.8
6.2
7.4
5.3
6.4
6.2
6.6
6.0
(continued)
61
-------
TABLE 17 (continued)
Site No. Description
A4 Roadside
A5 Swale
P-9 Woods
P-10 Woods
1-1,1-2.
1-3,1-4 Golf Course
2-1,2-2,
2-3,2-4 Woods
3-1,3-2 Lawn
4-1,4-2,
4-3 , 4-4 Woods
5—1 5—2
5-3^5-4' Roadside
6-1, 6-2 ......
6-3,6-4 Swale
7-1, 7-2 Roadside
8-1, 8-2 Woods
AS Woods
BS Wooda
CS Woods
Disturbed Fertilized pft
+ + 6.7
+ + 7.7
6.0
- 5.9
7.2,7.0*,
+ + 7.1,6.4
6.0,5.3,
- 5.7,6.2
-I- + 8.1,7.7
6.8,6.9,
6.9,6.8
6.4,6.7,
+ + 6.5,7.8
8.8,8.5
+ + 8.7,7.2
4- •*• 8.5,8.5
7.0,7.0
5.7
6.3
6.1
1
indicates no
indicates yes
pH values respective to Site No,
62
-------
Representatives of 52 genera of algae were identified from
153 surface samples of soils collected from the 51 sites in the
study area (Table 18). The majority of genera/ in both disturbed
and undisturbed soils,, were members of the Chlorophyta, while
only three genera belonged to the Euglenophyta. Chlamydomonas,
Chlorococcum, Oscillatoria, and Phormidium were ubiquitous, while
several algae (Anabaena, Gloeotrichia, Pyrobotrys, and Scytonema)
were found only in disturbed soils which had received applica-
tions of commercial fertilizer. Thirteen genera of algae (10
greens, 1 diatom, 2 euglenoids) were present only in undisturbed
soils. There was no uniformity in the distribution of algae,
even in soils which had been collected from similar sites.
Actually, there is little reason to believe that exact uniformity
of algal populations would occur in a particular soil. Thus, the
possible combinations of edaphic algal communities which may de-
.velop in a particular area seem to be astronomical. All algal
cells which reach a particular soil do not .survive, due to their
inabilities to adapt to changing physicochemical conditions or to
form resistant cells. Archibald (43), on the basis of surveys of
edaphic algae in closely spaced samples collected along tran-
sections, stated that there is no predictability in the distribu-
tion of soil microalgae. On the other hand, Arvik (50) reported
that it is probable that an association may be found between
edaphic algae and a given soil type. Exact correlations between
edaphic algae and specific soil parameters will probably become
TABLE 18. SUMMARY OP EDAPHIC ALGAL DISTRIBUTION
IN THE WOODLANDS ,'
. Disturbed-fertilized
Division
Chlorophyta
Cyanophyta
Chrysophyta
Euglenophyta
Woods
27
5
11
3
Disturbed-
unfertilized
15
5
7
1
Golf '
Course
7
4
4 '
1."
•Swa 1 e
'3
7
4
1
Road-
side
5
5
4
1
Lawn
4
4
3
—
Total genera/
site
46
28
16
15
15
11
Total genera identified—52
63
-------
evident only when the microalgae are identified to the species
level. Unfortunately, specific identifications of algae are
ofte^ laborious and time-consuming; thus, they are not feasible
when large numbers of soils are being analyzed. Even so,
identifications of edaphic algae to the generic level provide
information for determining Overall effects of land usage on
algal populations.
Undisturbed soils in the study area had a greater diversity
of algae, particularly green! algae, than the disturbed soils.
Disturbance of soils favored; the development of a more diverse
blue-green algal flora, probably due to increases in soil pH
which accompanied soil disturbance. As shown in Table 18, soil
disturbance affects the ratio of green to blue-green algae, with
the number of genera of blue-green algae exceeding the number of
green algae in the soils which had the highest pH values. With
decreasing pH, there was a tendency toward a dominance of green
algae. Floristic surveys in; other regions of the United States
(17,48,56) have also shown that alkaline soils favor the develop-
ment of a more luxuriant bluk-green algal flora than do acid
soils. I
I
The largest standing crbps of edaphic algae were found in
soils collected from the golf course (Table ;19). Green algae
were more numerous in soils with a pH of 7.3 or less, while
members of the Chrysophyta outnumbered blue-green algae regard-
less of soil pH. Jurgensen find Davey (41) have stated that an
inverse relationship exists between soil pH and algal numbers.
This relationship seems to apply to disturbed soils in the study
area, with the exception of jsoils from the roadside. Roadside
collection sites were located on steep inclines, and washout of
algae by surface runoff may have been responsible for reduced
standing crops at these sites. However, undisturbed soils had
the lowest pH and the smallest standing crop of algae. Thus, the
above relationship, if it is! valid, might apply only to soils
collected from similar sites: where pH is the major variable.
Standing crops of algae decreased with soil depth in the
study area (Table 20) . This! inverse relationship between algal
numbers and soil depth has a|lso been noted for other soils in
the United States (53,54), and surveys of vertical cores of soil
(52,55) indicate that algae are not usually encountered below
soil depths of 15 cm. This Was the case for two sites in the
study area, but at one site IChlamydomonas was found at depths of
30 cm. Soils from the latte|r site were water-logged for extended
periods, thus providing amplje moisture which perhaps facilitated
active movement of algae to [greater depths. Some algae may
actively migrate when soil mpisture is abundant, particularly if
they are facultative heterotbrophs. However, light availability
limits most algae to surface soils, and their presence in lower
strata is probably due to re'sistant cells which are moved by
64
-------
TABLE 19. ALGAL.NUMBERS (CELLS/G) IN VARIOUS
SOILS IN THE WOODLANDS
Collection
Site
Golf course
Lawn
Swale
Roadside
Woods
Chlorophyta
39,629*
17,400
8,298
6,628
3,853
Cyanophyta
1,446
7,195
4,533
1,788
586
Chrysophyta
13,084
12,790
22,238
6,264
1,297
Total
54,159
37,385
35,069
14,680
5,73.6
figures represent average values
TABLE 20. ALGAL NUMBERS (CELLS/G) AT VARIOUS DEPTH INTERVALS
IN CORE SAMPLES FROM THE WOODLANDS
Soil Depth (cm)
0 -
5 -
10 -
15 -
20 -
25 -
30-35
35 -
5
10
15
20
25
30
40
Collection Site*
AS
1,200+
1,600
100
0
0
0
0
0
BS
4,000
1,500
2,300
500
100
100
. 0
0
C.S
1,900
400
100
0
0
0
0
0
•c
refer to Figure 26
average of triplicate samples
65
-------
percolation of water. Bold ([personal communication) has found ;'•.
algae at depths of 244 cm in [soils near Austin, Texas.
Leachates from golf course soils had the highest concentra-
tions of nitrogen and phosphorus (Table 21) , due to regular ' •,
additions of commercial fertilizer (Scott's Proturf). The lawn
and roadside collection sites' were also fertilized, but not as
heavily as the golf course. JFertilizers reached the swale soils
only by way of surface runoff from fertilized areas, and decom-
position of plant litter was probably the major source of nitro-
gen and phosphorus for the swales. Likewise, decomposition of
plant debris was the primary isource of nitrogen and phosphorus
for soils in the undisturbed areas of the study area. In fact,
the wooded areas had a thick ground cover of plant debris, and
in many locations thick layers of humus were present. However>,
it seems that standing crops and diversity of edaphic algae are.
not dependent solely upon concentrations of nitrogen and phos- ••
phorus in the soils. Disturbance of soil and changes in pH, as
previously discussed, are probably primary factors which influ-,
ence algal distribution. For; example, the undisturbed soils had
a more diverse algal population but a smaller standing crop of
algae than the disturbed soils, even though concentrations of
nitrogen and phosphorus were higher than in most disturbed soils.
Admittedly, soil leachates do! not measure total nitrogen and
phosphorus in the soils, but rather quantities of nitrogen and
phosphorus which are readily released from soils.
TABLE 21. CHEMICAL 'ANALYSES OF LEACHATES FROM
VARIOUS SOILS IN THE WOODLANDS
i
Collection
Site
Golf course
Lawn
Swale
Roadside
Woods
pH
6
6
7
7
6
.7*
.9
.8
.3
.1
NO^-N
(mgj/1)
2.
84
0.68
0.
1.
1.
46
26
15
NO2-N
(mg/1)
0
0
0
0
0
.260
.008
.015
.016
.300
0-P04
(mg/1)
3
0
0
0
0
.42
.30
.87
.33
.99
T-P
(mg/1)
5
0
1
0
2
.41
.53
.00
.53
.11
average values
66
-------
In order to assess the role of undisturbed soils in provid-
ing nutrients for algal growth, bioassays were conducted with
leachates from three undisturbed site| •.:,;• Nitrogen and phosphorus
concentrations of leachates are presented in Table 22 and the
results of the bioassays are shown in Figure 28. The leachates
exhibited varying abilities to support growth of the test alga.
Growth was enhanced significantly by additions of phosphorus to
leachates from all sites, while addition of nitrogen did not
significantly influence the yield of the test alga. Thus, if
undisturbed areas are inundated, phosphorus could possibly be
the nutrient most limiting for algal growth. Bioassays with
water from the man-made lakes in the study area indicate that
phosphorus is the nutrient most limiting for algal growth. How-
ever, during periods of rainfall, significant quantities of
nutrients are washed into the lakes from surrounding lawns and
golf course greens. Bioassays with water collected from the
lakes during periods of surface runoff indicate that nitrogen is
the limiting nutrient.
TABLE 2'2. CHEMICAL ANALYSES OF LEACHATES FROM SURFACE SOILS
- UTILIZED IN ALGAL BIOASSAYS
Collection
Site*
AS
BS
CS
pH
6.4
6.9
6.6
NO3-N
(mg/1)
3.90
1.54
2.88
NO2-N
(mg/1)
0.50
0.96
0.56
0-P04
(mg/1)
0.22
0.30
0.40
refer to Figure 26
as
Since terrestrial and aquatic ecosystems are connected
hydrologically, they cannot be considered as totally separate
units. Surface drainage serves as a major component in this
hydrologic linkage and is thus an important ecological para-
meter. Land usage in a watershed will determine the quality,
well as the quantity, of surface runoff; thus, influencing '
aquatic habitats in the watershed. In addition to providing
potential nutrients for algal growth, surface drainage probably
transports algal cells to aquatic habitats from surrounding
soils. Since land use affects edaphic algal populations, it
must also influence the diversity of algae which could be trans-
ported by surface runoff. For example, a more diverse assemblage
of blue-green algae could potentially enter lakes in the study
area by surface drainage from disturbed rather than undisturbed
soils.
67
-------
0.60
0.50
UJ 0.40
Q
0.30
0.20
0.10
<
o
Q_
O
I-LEAQH ATE-SPIKES
2-LEAdHATEH.OpfJM
3- LEACHATE4- 0.05ppMP
3
1
in
AS '> BS CS
COLLECTION SITES
Figure 28. Growth of Selenastrum in
leacbhates of various soils
from The Woodlands.
68
-------
AQUATIC VASCULAR PLANTS
^ aquatic vascular plants of The Woodlands were not ex-
tensively investigated due to their limited distribution in the
watershed. Only 15 species of aquatic macrophytes were identi-
fied^ and these were found in only four of the established col-
lection sites. Of these species, only three were found in the
Conference Center Lakes.
During the first year of study, the Conference Center Lakes
did not develop a population of aquatic vascular plants, but
during the second year, Juncus rep ens, and Sagittaria graminea
invaded the southwestern side of Lake B. However, in the third
year of study, the major portion of the littoral zone of Lake B
and portions of Lake A (Figure .29) developed rather extensive
stands of Typha latifola, in addition to the two species
mentioned above. Thus, in a three-year period, the aquatic
vascular plants have invaded and spread through the littoral
zone of these lakes. Unless some management strategy is insti-
tuted, there is a strong probability that these plants will
totally encompass the lakes and gradually develop to nuisance
levels.
69
-------
LAKE A
X - Location of
Aquatic Vasciklar
Plants !
LAKE B
Figure 29.
Distribution of aquatic vascular
plants in the Conference Center
Lakes.
70
-------
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75
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APPENDIX
METHODS; AND PROCEDURES
i
ROUTINE PROCEDURES i
«*
Standard research techniques were utilized in most phases of
this investigation. However[, detailed descriptions for routine
laboratory procedures, such as cleaning and sterilization of
material, filtering procedures, and organism transfer techniques,
will not be discussed due to universality of their use.
LIST OF BIOLOGICAL METHODS AND MEDIA
Method j Reference
. — I .
Quantitative Algal Samples | 1
Sample Concentration 1
Algal Enumeration ; 1
Sample Preservation i 1
Algal Identifications 1-12
Aquatic Macrophyte Identification 15, 16
Hold's Basal Medium 18
Edaphic Algal Isolation 17
Knop's Medium 17
Aspiration Procedure 19
Bioassay Procedure ; 13
Growth Measurements 13, 14
Standard Test Medium ; . 13
Standard Test Algae 13
76
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ALGAL PROCEDURES
Water samples for algal studies were collected from 15
sites in The Woodlands area. The samples were collected in ac-
cordance with procedures in Standard Methods (1) and were
processed immediately upon return to the laboratory. Water
samples for algal enumeration were collected in pre-cleaned, 3.7
liter glass bottles. Algal samples were concentrated in a
Sedgwick-Rafter column (1) and algal enumeration was facilitated
by use of a Whipple disc and Sedgwick-Rafter counting cell (1).
When possible, at least one liter of water was concentrated to a
final volume of 5 ml. Concentrated samples were preserved with
Lugol's iodine (1) and each individual alga was counted as one
unit. ' • '
Samples for phytoplankton identification were collected by
making several horizontal hauls at each station with a plankton
net (#25 mesh). Preservatives were not added to these samples.
Algal species were identified by microscopic examinations of wet
mounts of material and each sample was examined until no addi-
tional algae were observed. A variety of taxonomic references
(1-12) were consulted for species identification.
ALGAL BIOASSAYS
Water samples for use in bioassays were also collected in
pre-cleaned, 3.7 liter glass bottles. Bioassays were conducted
according to the procedures in the EPA "Algal Assay Procedure
Bottle Test" (13). Cotton plugged, 125 ml Erlenmeyer flasks were
used as culture containers and all experiments were, run under
standard culture conditions. In all cases, 40 ml of treated
water sample were used and every effort was expended to maintain
sterile conditions in test preparation. Optical densities
(650 nm) and/or dry weight determinations (14) were made at
appropriate intervals during the incubation period.
The storm event samples were filtered through 0.45 y filters
and were spiked with various concentrations of nitrate-nitrogen
(NaN03) and orthophosphate-phosphorus (K2HPO4). Sample pH was
adjusted to 7 and the flasks were inoculated with Selenastrum
capricornutum. Samples were incubated and growth measurements
were made at appropriate time intervals. Additional samples
from a Hunting Bayou storm event were similarly prepared and
inoculated with Anabaena flos-aquae.
The majority of the stream low-flow samples were treated as
described above. However, on one occasion, aliquots of a sample
were autoclaved and inoculated with S_. capr icornutum. Other
aliquots of the same sample were filtered through glass, 1.2 y
and 0.45 p filters. The glass and 1.2 u filtered samples were
autoclaved before inoculation, while the 0.45 y filtered aliquots
77
-------
were inoculated directly. The pH of some of the 0.45 y filtered
samples was adjusted before [inoculation.
The standard EPA test medium (13) was used to maintain the
inocula for these bioassays.i Variations of this medium were
also used as a test medium in conjunction with the collected
water samples. ;
AQUATIC VASCULAR PLANTS [
!
Aquatic vascular plants | were collected by hand from the
Conference Center Lakes and wet weather ponds in The Woodlands
area. Collections were transported to the laboratory for sorting
and identification. Species5 identifications were made by use of
standard taxonomic references (15,16).
EDAPHIC ALGAL POPULATIONS |
Fifty-one sites were selected for collection of surface soil
samples. The samples were taken from the lake basins before
inundation, and triplicate samples were collected at each site.
Soil samples were collected by use of a small garden trowel and
were placed in sterile plastic bags for transport to the labora-
tory. To minimize carry-over from sample to sample, the trowel
was wiped clean, immersed in alcohol and flamed after each
sampling. In the laboratory^ each sample was divided aseptically
into four 20 g portions. Two of the portions were placed in
separate 125 ml Erlenmeyer flasks containing 100 ml of sterile
Knop's Medium (17), while the remaining two were placed in simi-
lar flasks of sterile Hold's;Basal Medium (BBM) (18). The pre-
pared flasks were placed in a controlled environment culture
chamber at a temperature of 23 ± 1°C. An incident light of
400 ft-c was provided by 40 vj? cool-white fluorescent light bulbs
set on a 12-hr light, 12-hr dark cycle. These conditions will
hereinafter be referred to as standard culture conditions.
After two weeks, the contents of each flask were examined
microscopically and the algal genera were tabulated. The obser-
vations were repeated every week for a period of six weeks.
Samples of algae were also removed from each flask and suspended
in separate tubes of liquid BBM. The contents of each tube were
aspirated (19) over duplicate petri dishes containing sterile
BBM solidified with 15 g agar. The seeded dishes were then
incubated under standard culture conditions. Upon examining the
agar surfaces at two weeks, dissimilarities in colonial morphol-
ogy were noticed. Portions of the various colonies were removed
from the agar surface by meaijis of sterile Pasteur pipettes,
drawn to fine bores in a microflame, and placed in culture tubes
containing sterile liquid BBty. When growth appeared in these
tubes, the algae were studied, and identified to the generic
level.
78
-------
Algal numbers were determined by placing 2 g of soil in
100 ml of sterile liquid BBM. The contents of the flasks were
mixed thoroughly and 1 ml was pipetted aseptically onto the sur-
face of sterile BBM agar. The inoculum was spread with a sterile
transfer loop and the plates were incubated under standard cul-
ture conditions. After ten days, the number of colonies belong-
ing to the Chlorophyta, Cyanophyta, and Chrysophyta were deter-
mined with the aid of a dissecting microscope. Triplicate plates
were used for each soil tested.
Soil leachates, for use in chemical analyses and algal bio-
assays, were obtained by placing 250 g of oven-dried soil in
vertical, plexiglass columns (length, 75 cm; internal diameter,
4.5 cm) which had their lower ends plugged with rubber stoppers.
Each stopper was equipped with a glass tube to allow for passage
of effluent water. Sterile glass wool was placed between the
stopper and soil to permit movement of water without loss of
soil from the column. Once the column had been prepared, 100 ml
of deionized water were placed over the soil. The first 100 ml
of effluent water were discarded and the next 500 ml were col-
lected in pre-cleaned glass bottles. The effluent was filtered
through prewashed, 0.45 y Gelman filters, and portions of the
filtrates were analyzed for N03-N, NO2-N, O-PO4, and total phos-
phorus with a Technicon Autoanalyzer.
Soil samples were collected from various depths by use of a
metal core sampler (length, 70 cm; Internal diameter, 4.5 cm).
Two-inch sections were removed from the collected soil column
and each was wrapped in a sterile plastic bag. The bore of the
core sampler was swabbed between collections to prevent carry-
over of organisms from sample to sample. In the laboratory, the
outer layers of each sample were removed and discarded. The
remaining soil was mixed thoroughly and algal numbers were deter-
mined by the procedures outlined above. Triplicate core samples
were collected at each of three collection sites.
Methods used in, the batch culture bioassays with filtrates
of soil leachates were similar to those described in "Algal Assay
Procedure Bottle Test" (13) . Forty-milliliter .portions of each
filtrate were transferred to sterile 125 ml Erlenmeyer flasks.
Three flasks were used as controls (no added nutrients), while
other sets of three flasks were spiked with nitrogen (1 mg/1 N as
NaNOs), and/or phosphorus (0.05 mg/1 P as KH2P04). The prepared
flasks were seeded with Selenastrum capricornutum (EPA culture),
and initial cell concentrations were adjusted so that each flask
contained 1 x 103 cells/ml. Cotton plugs were used as closures
for the flasks and each flask was shaken daily for mixing. Algal
growth was determined with a Beckman Spectrophotometer (650 nm)
after 21 days incubation.
79
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WATER CHEMISTRY
I
All chemical parameters iwere measured in the Water Chemistry
Laboratory, Rice University, junder the direction of Dr. F. L.
Roe.
80
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APPENDIX
REFERENCES
1. Standard Methods for the Examination of Water and Waste-
water, 13th ed. A.P.H.A., W.P.C.F., A.W.W.A., 1971.
2. Huber-Pestalozzi, G. Das Phytoplankton des Susswassers.
In: Die Binnengewasser, A. Thienemann, ed. Schweizer-
bart'sche Verlazsbuchhandlunz, Stuttgart, 1938-1961.
3. Patrick, R., and C. W. Reimer. The Diatoms of the United
States, Exclusive of Alaska and Hawaii, Vol. 1, Monograph
No. 13. Acad. Nat. Sci., Philadelphia, Pennsylvania,
1966. 688 pp.
4. Prescott, G. W. Algae of the Western Great Lakes Area,
revised ed. William C. Brown Co., Publishers, Dubuque,
1962. 977 pp.
5. Smith, G. M. Phytoplankton of the Inland Lakes of Wiscon-
sin, Part I. Wisconsin Geol. Nat. Hist. Surv. Bull. No.
57. Madison, 1920. 243 pp.
6. Smith, G. M. Phytoplankton of the Inland Lakes of Wiscon-
sin, Part II. Wisconsin Geol. Nat. Hist. Surv. Bull. No. ,
57. Madison, 1924. 227 pp.
7. Smith, G. M. The Freshwater Algae of the United States,
2nd ed. McGraw-Hill Book Co., Inc., New York, 1950.
719 pp.
8. West, G. S., and W. West. A Monograph of the British
Desmidiaceae, Vols. 1-4. The Roy. Soc., London, 1904-1912.
9. West, G. S., W. West, and N. Carter., A Monograph of the
British Desmidiaceae, Vol. 5. The Roy. Soc., London, 1923.
10. West, G. S., and F. E. Fritsch. A Treatise on the British
Freshwater Algae. Bibliotheca Phycologica, Ban 3, 1968.
534 pp.
11. Tiffany, L. H., and M. E. Britton. The Algae of Illinois.
University of Chicago Press, Chicago, 1952. 407 pp.
81
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12. Gomont, M. Monographic Des Oscillariees. Annales des
Sciences Naturelles, Vlli, 15:256-263, 1892.
13. Algal Assay Procedure Bottle Test. National Eutrophication
Research Program, U.S. Environmental Protection Agency,
August, 1971. 82 pp. '•
14. Sorokin, C. Dry Weight,; Packed Cell Volume and Optical
Density. In: Handbook of Phycological Methods, J. R.
Stein, ed. Cambridge University Press, Oxford, 1973.
448 pp. I
15. Correll, D. S., and H. BJ. Correll. Aquatic and Wetland
Plants of Southwestern United States. Water Pollution
Control Research Series,( No. 1630 DNL 01/72, U.S. Environ-
mental Protection Agency;, 1972. 1777 pp.
16. Hotchkiss, N. Underwater and Floating-Leaved Plants of the
United States and Canada1. Resource Publication 44. U.S.
Department of Interior, bureau of Sport Fisheries and Wild-
life, Washington, D.C., [1967.
17. MacEntee, F. J. A Preliminary Investigation of the Soil
Algae of Northeastern Pennsylvania. Soil Science, 110:
313-317, 1970. |
18. Nichols, H. W. Growth Media - Freshwater. In: Handbook of
Phycological Methods, J.! R. Stein, ed. Cambridge University
Press, Oxford, 1973. 448 pp.
19. Hoshaw, R. W. , and J. R.| Rosowski. Methods for Microscopic
Algae. In: Handbook of Phycological Methods, J. R. Stein,
ed. Cambridge University Press, Oxford, 1973. 448 pp.
82
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/2-80-127
2.
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
MAXIMUM UTILIZATION OF WATER RESOURCES IN A PLANNED
COMMUNITY; Eutrophication Potential of Surface
Waters in a Developing Watershed
5. REPORT DATE
August 1980 (Issuing Date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
8. PERFORMING ORGANIZATION REPORT NO,
C. H. Ward and J. M. King
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Department of Environmental Science & Engineering
Rice University
P.O. Box 1892
Houston, Texas 77001
10. PROGRAM ELEMENT NO.
35B1C,DU No.B-124,Task 404202
11. CONTHACT/GRANT NO.
802433
12. SPONSORING AGENCY NAME AND ADDRESS
Municipal Environmental Research Laboratory—Cin.
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio
~,OH
13. TYPE OF REPORT AND PERIOD COVERED
Final 7/73 - 12/76
14. SPONSORING AGENCY CODE
EPA/600/14
is. SUPPLEMENTARY
Project Officers:
al so EPA-600/2-79-050a , b , c , d ,e , f and EPA-600/2-80-113
Richard Field (201) 321-6674 - FTS 3-340-6674
Anthony N. Tafuri (201) 321-6676 - FTS 8-340-6676 _
The purpose of this research was to characterize the algal populations in a
developing area (The Woodlands) to evaluate the impact of urbanization on the aquatic
flora in The Woodlands. Several aquatic habitats were sampled on a regular basis to
identify factors which influence algal population dynamics. Nutrient limitation studie<
were conducted to determine which nutrient was most limiting for algal growth during
conditions of low flow and stormwater runoff. Water from Hunting Bayou and Westbury
Square, developed communities near Houston, Texas, wer.e used in bioassay experiments
The impact of urbanization on edaphic algal populations " ' "
limitation studies in Panther Branch and the Conference
phorus additions to low-flow water increased algal cell
water samples were increased by nitrogen additions.
was also determined. Nutrient
Center Lakes showed that phos-
yields, while yields in stomm-
Undisturbed soils had more diverse algal populations, but smaller standing crops,
than disturbed soils, even though concentrations of nitrogen and phosphorus were higher
than in most disturbed soils. Soil disturbance caused development of a more diverse
blue-green algal flora, probably due to accompanying increases in soil pH. Bioassays
showed that phosphorus was the nutrient most limiting for algal growth in water
leachates of soils.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS
c. cos AT I Field/Group
Water resources, Urbanization
Aquatic flora, Nutrient
limitation data, Algal
growth, Nutrient loadings
Water leachates, Edaphic
algae, Phytoplankton
dynamics, Algal bioassays;
Eutrophication i
13B
18. DISTRIBUTION STATEMENT
Release to public
19. SECURITY CLASS (ThisReport)
Unclassified
21. NO. OF PAGES
97
20. SECURITY CLASS (Thispage)
Unclassified
22. PRICE
EPA Form 2220-1 (Rev. 4-77)
•fr U.S. GOVERNMENT PRINTING OFFICE: 1980--657-165/OH7
83
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