PB83-136341
Community Structure, Nutrient Dynamics, and the
Degradation of Diethyl Phthalate in
Aquatic Laboratory Microcosms
(U.S.) Environmental Research Lab.
Athens, GA
Nov 82
U.S. DEPARTMENT OF COMMERCE
National Technical Information Service
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EPA-600/3-82-093
November 1982
COMMUNITY STRUCTURE, NUTRIENT DYNAMICS, AND THE
DEGRADATION OF DIETHYL PHTHALATE IN AQUATIC LABORATORY MICROCOSMS
by
T. L. Hall, J. A. Hamala, P. F. Hendrix, *H. P. Kollig,
J. A. Krewer, C. L. Langner, and *W. R. Payne, Jr.
The Bionetics Corporation
Environmental Research Laboratory
College Station Road
Athens, Georgia 30613
*U.S. Environmental Protection Agency
Environmental Research Laboratory
College Station Road
Athens, Georgia 30613
ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
ATHENS, GEORGIA 30613
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-82-093
3. RECIPIENT'S ACCESSIOC*NO.
5 13634 1
4. TITLE AND SUBTITLE
Community Structure, Nutrient Dynamics and the Degrada-
tion of Diethyl Phthalate in Aquatic Laboratory
Microcosms
5. REPORT DATE
November 1982
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
T.L. Hall, J.A. Hamala, P.P. Hendrix, H.P. Kollig,
J.A. Krewer, C.L. Langner, and W.R. Payne, Jr.
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
The Bionetics Corporation and Environmental Research
Laboratory
U.S. Environmental Protection Agency
Athens, Georgia 30613
10. PROGRAM ELEMENT NO.
ACUL1A
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Research Laboratory—Athens, GA
Office of Research and Development
U.S. Environmental Protection Agency
Athens, Georgia 30613
13. TYPE OF REPORT AND PERIOD COVERED
Final, 11/79-1/81
14. SPONSORING AGENCY CODE
EPA/600/01
15. SUPPLEMENTARY NOTES
16. ABSTRACT
An investigation was conducted of the environmental fate of diethyl phthalate
(DEP) in the continuous-flow channel microcosms housed in the USEPA's Environmental
Research Laboratory, Athens, Ga. The objectives of the investigations were to deter-
mine (1) whether a definable stable state could be achieved, (2) the effects of dif-
ferent nutrient treatments on ecosystem structure and function and on the fate of DEP
and (3) the degree of similarity between replicate microcosms.
Aufwuchs assemblages in the microcosms reached fairly stable levels of biomass
metabolic activity, and similar species composition within 2 or 3 months after inocu-
lation. Communities receiving direct nutrient inputs appeared to stabilize first,
followed by downstream communities. A highly significant relationship between phos-
phorus inputs and aufwuchs chlorophyll a_ was established suggesting that the relative
ly stable input concentrations of inorganic nutrients into any given compartment were
among the primary factors controlling maximum development of aufwuchs.
Replicate microcosms were statistically indistinguishable with respect to nu-
trient concentrations for most of the experimental period. Compartments receiving
direct inputs of inorganic nutrients had the most consistent replicability. Although
non-taxonomic community structure was generally similar in replicate compartments,
some differences were observed in relative species abundance. Sorption, volatiliza-
tion, and photolysis were insiginficant processes in the fate of DEP.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
13. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (This Report)
UNCLASSIFIED '
21. NO. OF PAGES
148
20. SECURITY CLASS (This page)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
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DISCLAIMER
This report has been reviewed by the Environmental Research Laboratory,
U.S. Environmental Protection Agency, and approved for publication. Mention
of trade names or commerical products does not constitute endorsement or
recommendation for use.
ii
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E'ORE~'JORD
Environmental protection efforts are increasingly directed towards pre-
venting adverse health and ecological effects associated with specific com-
pounds of natural or human origin. As part of this Laboratory's research on
the occurrence, movement, transformation, impact, and control of environmen-
tal contaminants, the Environmental Systems Branch studies complexes o'f
environmental processes that control the transport, transformation, degrada-
tion, and impact of pollutants or other materials in soil and water and
assesses environmental factors that affect water quality.
Concern about environmental exposure to toxic substances has increased
the need for accurate information on the transport, fate, and effects of
trace contaminants in natural waters. One technique that shows promise as a
useful tool for providing some of this information is the use of laboratory
microcosms as ecosystems for the assessment of pollutant exposure to natural
aquatic systems. This report presents an evaluation of microcosms as :est
systems for exposure analysis models using a phthalate ester as the study
compound.
David W. Duttweiler
Director
Environmental Research
Athens, Georgia
LcboratoL:Y
iii
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ABSTRACT
The research reported herein was conducted in two 8~compartment continu~
ous~flow channel microcosms housed in the USEPA's Environmental Research
Laboratory, Athens, Georgia. The microcosms were designed and established
to enable testing of the Exposure Analysis Modeling System (EXAMS)~ a theo~
retical~type predictive model for the determination of the fate of toxic
compounds in freshwater systems.
The objectives of the investigations were to determine (1) whether a
definable stable state could be achieved in which to test the model; (2) the
degree of similarity between corresponding microcosms: (3) the effects of
different nutrient treatments on ecosystem structure and function: and (4)
the fate of diethyl phthalate (DEP) in channel microcosms.
Aufwuchs assemblages in the microcosms reached fairly stable levels of
biomass, metabolic activity~ and similar species composition within two or
three months after inoculation. Communities receiving direct nutrient
inputs appeared to stabilize first~ followed by downstream communities.
Replicate microcosms were statistically indistinguishable with respect
to nutrient concentrations for most of the experimental period. Compart~
ments receiving direct inputs of inorganic nutrients had the most consistent
replicability. Although non~taxonomic community structure was generally
similar in replicate compartments, some differences were observed in relae
tive species abundance.
A highly significant relationship between phosphorus inputs and aufwuchs
chlorophyll a was established suggesting that the relatively stable input.
concentrations of inorganic nutrients into any given compartment were among-
the primary factors controlling maximum development of aufwuchs. ~~
Sorption~ volatilization~ and photolysis were insignificant processes in
the fate of DEP. Alkaline hydrolysis at pH 10 showed only a slight effect.
Microbial degradation was the dominant process. First~order degradation.
rates were all within an order of magnitude even though there were signifie
cant differences in both chemical environments and biological communities.
This report covers the period November 13, 1979, to January 29, 1981,
and work was completed as of December 1981.
iv
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CONTENTS
For eword . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . . . . . . .
Abstract[[[
Figures............................ 0.......,...................,..'.....'
Tables[[[ .
ACknowledgments.......................................0..................
Literature
1.
2.
3.
4.
5.
6.
7.
Introduction.........................o........................~.
Conclusions[[[
Recommendations.................................................
Facility[[[
Materials and Methods...........................................
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Number
FIGURES
1
Toxicant Feed System.............................................
3
Orthophosphate Concentrations in CSTRs A3
Experimental Conditions of Each Channel..........................
2
4
Orthophosphate Concentrations in CSTRs A4
S
Orthophosphate Concentrations in CSTRs AS
6
Orthophosphate Concentrations in CSTRs A6
7
Orthophosphate Concentrations in CSTRs A7
8
Orthophosphate Concentrations in CSTRs AS
9
and B3........0........
and B4.................
and B5.................
and B6.................
and B7.............~...
and B8.................
10
Total Dissolved Phosphorus Concentrations in CSTRs A3 and B3.....
Total Dissolved Phosphorus Concentrations in CSTRs A4 and B4.....
11
Total Dissolved Phosphorus Concentrations in CSTRs AS and B5.....
12
Total Dissolved Phosphorus Concentrations in CSTRs A6 and B6.....
13
Total Dissolved Phosphorus Concentrations in CSTRs A7 and B7.....
14
15
Nitrate-Nitrogen Concentrations in CSTRs A3
Total Dissolved Phosphorus Concentrations in CSTRs A8 and B8.....
16
Nitrate-Nitrogen Concentrations in CSTRs A4
17
Nitrate-Nitrogen Concentrations in CSTRs AS
18
Nitrate-Nitrogen Concentrations in CSTRs A6
19
Nitrate-Nitrogen Concentrations in CSTRs A7
20
Nitrate-Nitrogen Concentrations in CSTRs A8
21
Nitrite-Nitrogen Concentrations in CSTRs A6
22
Nitrite-Nitrogen Concentrations in CSTRs A7
23
Nitrite-Nitrogen Concentrations in CSTRs A8
24
Ammonia-Nitrogen Concentrations in CSTRs A3
vi
and B3...............
and B4...............
and B5...............
and B6...............
and B7...............
and B8...............
and B6...............
and B7...............
and B8...............
and B3...............
Page
38
39
40
41
42
43
44
45
46
47
4~
49
so
51
52
53
54
55
56
57
58
59
60
61
-------
25
Ammonia-Nitrogen Concentrations in CSTRs A4
and 84...............
26
Ammonia-Nitrogen Concentrations in CSTRs A5
and 85.........0.....
27
Ammonia-Nitrogen Concentrations in CSTRs A6
and B6...............
28
Ammonia-Nitrogen Concentrations in CSTRs A7
and 87...............
29
Ammonia-Nitrogen Concentrations in CSTRs AS
and B8....0..........
30
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A3 and B3....
31
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A4 and 84....
32
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A5
and 85....
33
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A6 and B6....
34
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A7 and B7....
35
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A8 and B8....
36
Dissolved Organic Carbon Concentrations in CSTRs A3
and B3.......
37
Dissolved Organic Carbon Concentrations in CSTRs A4 and 84.......
38
Dissolved Organic Carbon Concentrations in CSTRs A5
and B5.......
39
Dissolved Organic Carbon Concentrations in CSTRs A5
and B6.......
40
Dissolved Organic carbon Concentrations in CSTRs A7
and 87.......
41
Dissolved' Organic Carbon Concentrations in CSTRs AS and B8.......
42
Total Organic Carbon Concentrations in CSTRs A3
and B3...........
43
Total Organic Carbon Concentrations in CSTRs A4
and B4...........
44
Total Organic Carbon Concentrations in CSTRs A5
and B5...........
45
Total Organic Carbon Concentrations in CSTRs A6 and B6...........
46
Total Organic. Carbon Concentrations in CSTRs A7
and B7...........
47
Total Organic Carbon Concentrations in CSTRs A8
and B8...........
48
Dissolved Oxygen Concentrations in A3
and 83.....................
49
Dissolved Oxygen Concentrations in A4
and 84.....................
50
Dissolved Oxygen Concentrations in A5
and 85.....................
vii
.- ..- ..' .. ,-.-_.-...,.,..--'-.,.., .'~ ~-...- --, ...._~-. .
62
63
64
6S
66
67
68
69
70
71
72
73
74
75
76
77
78
79
~o
81
82
~3
84
8S
86
87
-------
51
Dissolved Oxygen Concentrations in A6
and B6..'..........8.......
52
Dissolved Oxygen Concentrations in A7
and B7'O............. 0 . . . . . .
53
Dissolved Oxygen Concentrations in A8
and Ba.....................
54
pH Values in CSTRs A3
and B3......................0..............
55
pH Values in CSTRs A4
an d 84....................... II 0 . . 0 . . . . :a 8 . . .
56
pH Values in CSTRs A5
and 85..............04... 0 .00.0 . . 0 . . . 0 . . . . . .t
57
pH Values in CSTRs A6
an d B 6 . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0 . . . 0 . 0 . .
58
pH Values in CSTRs A7
and B 7 . -0 . 0 . . . . . . . . . . . . . . 0 . .. . . . . .. ~ . . II . . . . . . .
59
pH Values in CSTRs A8
an d B8............................. 0 0 . . . . . 0
60
Aufwuchs chlorophyll a in AEcoS at three sampling times
corresponding to 2 months (early), 6 months (mid), and 9
months (late) of system development.s..........s.................
61
Aufwuchs adenosine triphosphate in AEcoS at three sampling
times corresponding to 2 months (early), 6 months (mid),
and 9 months (late) of system development..~oo..4.~..~..~oo...,~.~
62
AUfwuchs ash-free dry weight in AEcoS at thcee sampling
times correspondi!'.g to 2 months (early), 6 months (mid),
and 9 months (late) of system development........................
63
Aufwuchs total organic carbon in 1l.EcoS at three sampling
times corresponding to 2 months (early); 5 months. (mi.d),
and 9 months (la.te) of syst2m dEvelopment........~...............
64
Aufwuchs bacter ia1 numbers in AEcoS at three s.::.mpling
times corresponding to 2 months (early), 6 months (mid),
and 9 months (late) of system development........................
65
First order rates of degradation (Kl) for DE? in AEcoS
(Channels A and B averaged) with 95% confidence intervals........
66
Orthophosphate Output:lnput Ratios
in CSTP.s fl.3
and B3o..o..~O.O.9
67
Orthophosphate Output::nput
Ratios in
CSTRs A6
~ ,....r
ana ~OQ~..oo~o.eo.
6'8
Nitrate-Nitrogen Output:Input Ratios
in CSTRs A3
and B3..........
69
Nitrate-Nitrogen Output:Input Ratios in CSTRs A6
and 86..........
70
Ammonia-Nitrogen Output:Input Ratios in CSTRs A3
and B3..........
71
Ammonia-Nitrogen Output:Input Ratios in CSTRs A6
and B6..........
viii
88
09
90
91
92
93
94
95
96
97
93
99
100
101
102
10.3
104
105
106
107
108
-------
72
Linear relationship between aufwuchs chlorophyll a concentra-
tion and total dissolved phosphorus input concentration for
Channel A, Channel B, and Channels A and B combined..............
73
Curvilinear relationship between aufwuchs chlorophyll a concen-
tration and total dissolved phosphorus input concentration for
Channels A and B combined........................................
74
Dissolved Nitrogen:Phosphorus Output Ratios in CSTRs A3 and B3...
75
Dissolved Nitrogen:phosphorus Output Ratios in CSTRs A6 and B6...
l.X .
109
110
III
112
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Number
TABLES
1
Elemental concentrations in
AEcoS...............................
2
Nutrient quantities in 44 1 of macronutrient concentrate
(drip rate of 4.0 ml/min).......................................
3
Nutrient quantities in 1.0 1 of micronutrient concentrate.......
4
Major sampling dates for DEP
experiment.........................
5
Steady-state concentrations of DEP (ug/l) due to sorption, photoly-
sis, and hydrolysis of a mean input concentration of
191 ug/l DEP (s = 6.0, n = 20).................................. 117
6
Concentrations (mg/l), percent degradation, and first-order
rate coefficients (kl) of DEP in AEcoS..........................
7
Estimated chlorophyll a, ATP, and bacteria for CSTRs 4 and 8
in sediments, suspended matter, and aufwuchs during DEP
additions. Numbers in parentheses indicate percentage of
total found in each phase.......................................
8
Calculated SIMI values for aufwuchs algal assemblages during
DEP additions...........................................08......
9
Groups of CSTRs having similar aufwuchs algal assemblages
during DEP additions - Channel A................................ 121
Groups of CSTRs having similar aufwuchs algal assemblages
during DE? addi tions - Ch ann el B................................ 122
10
11
Relative abundance (%) of aufwuchs algal species comprising
> 1.0% of the total density on 1/22/80..........................
12
Relative abundance (%) of aufwuchs algal species comprising
> 1.0% of the total density during DEP experimentation (8/5 -
9/16/80)[[[
13
Calculated HI diversity and J evenness values for aufwuchs
algal assemblages during DEP additions..........................
14
Results of Newman-Keu1s Range Test for aufwuchs biomass during
DEP additions (a = 0.05). Means expressed as 109l0(x + 1),
n = 6[[[
15
SIMI values comparing aufwuchs algal assemblages on 1/22/80 to
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16
Calculated product moment correlation coefficients for aufwuchs
structural parameters during DEP additions......................
17
Concentrations of trace metals in aufwuchs from AEcoS the week
of December 2, 1980.............................................
18
Chemical concentrations (mg/l) in CSTRs on major sampling dates
(see Table 4)...........o~....o.................................
19
Total dissolved phosphorus and aufwuchs chlorophyll ~
concentrations for each CSTR in AEcoS...........................
20
Analyzed mean N:P input and output ratios (by weight) for
11/16/79 - 12/1/80..............................................
21
Rep1icability and diel pattern Student's t-test results.........
22
Results of Student's t-test comparing aufwuchs biomass
parameters. in Channels A versus B during DEP additions..........
23
SIMI values comparing aufwuchs algal species composition
in Channels A and B during DEP additions........................
xi
128
129
130
131
132
133
134
135
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ACKNOWLEDGMENTS
We wish to thank the staff of the Environmental Systems Branch and The
Bionetics Corporation for their assistance, especially Harvey Holm of the
EPA during the planning stages. Technical support from Jacquelyn Benner and
David Lewis of the EPA was essential for completing diethyl phthalate analy-
ses and measuring microbial degradation rates. Biological analyses of the
water, sediment, and aufwuchs were completed through the assistance of R.
Vince Howard and Lita Proctor of The Bionetics Corporation.
Special thanks are due Ronnie Moon and Rudy parrish of the Computer
Sciences Corporation for providing statistical analyses and preparing
graphs, and to Sereta Wallis of EPA for the many hours of correcting and
typing the manuscript.
xii
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SECTION 1
INTRODUCTION
The EXposure Analysis Modeling System (EXAMS) was developed by the
Environmental Systems Branch of the USEPA's Environmental Research Labora-
tory, Athens, Georgia, to provide a tool for predicting the fate of chemical
compounds in natural waters (Burns et ale 1981). Since its implementation, .
the model has been tested with the chemical methyl parathion under con-
trolled laboratory conditions using the Aquatic Ecosystem Simulator (AECOS),
housed at the same laboratory. During 1980, the model was subjected to
another experimental test, again in AEcoS, using the chemical diethyl phtha-
late (DEP). The purpose of the present report is to (1) present the experi-
mental results of the DEP fate study in AEcoS, and (2) evaluate the degree
to which the experiments met some of the assumptions implicit in the EXAMS-
AEcoS testing procedure. EXAMS modeling results will be presented elsewhere.
-,.
ENVIRONMENTAL FATE OF DEP
Phthalic acid esters (PAEs) are plasticizers which are in widespread use -
today with approximately two million tons produced annually (Thomas et ale
1978). As plasticizers, PAEs are used in'PVC plastics, resulting in mate-
rials that are flexible and supple. PVC plastics are widely used for wire
and cable insulation, construction materials, home furnishings, automotive
components, apparel, and medical products. Non-plasticizer uses of phtha-
lates include pesticide carriers, munitions, cosmetics, and insect repel-
lents. Phthalate plasticizers are currently being produced and used in
amounts that could easily contribute to environmental pollution. The high
disposal rates of plastics may be producing an environmental reservoir of
the phthalates that may slowly leach into aquatic ecosystems (Giam et ale
1978). The two most commonly used phthalate compounds are di-n-butyl phtha-
late (DBP) and di(2-ethylhexyl) phthalate (DEHP). In a recent comparison of
concentrations of selected organics in the atmosphere (Atlas and Giam 1981),
both DBP and DEHP were found in the atmosphere in amounts above those found
for PCB, DDE, lindane, dieldrin, and chlordane.
Limited information exists on the fate of phthalate esters in biological
systems or aquatic environments despite their widespread use. PAEs have
been identified in soil (Ogner and Schnitzer 1970), fish (Mayer et ale
1972), food and biological samples (Cerbulis and Ard 1967, zitko 1973),
freshwaters of the Mississippi River and marine coastal waters of the Gulf
of Mexico (Corcoran 1973), open-ocean atmospheric samples of the Gulf of
Mexico (Giam et ale 1980), and waters of the Charles and Merrimack Rivers
(Hites 1973). Metcalf et ale (1973) studied the uptake of DEHP by a variety
of aquatic organisms using a laboratory model ecosystem with a terrestrial-
aquatic interface. This study showed that DEHPis rapidly accumulated by a
variety of aquatic plants and animals but is biodegraded very slowly by
algae, Daphnia~ snails and clams, and more rapidly in fish. The authors
suggest that the major degradative pathways are through hydrolysis of the
ester groups to produce monoethylhexyl phthalate, phthalic acid, and phtha-
lic anhydride. In support of this, Stalling et ale (1973) have shown that
DEHP is metabolized primarily to the mono ester in cha~nel catfish.
1
. ~ 0.-- - --i"-...~
-------
Several reports examine the biodegradation of phthalate esters by acti-
vated sludge and bacteria. Saeger and Tucker (1976) indicated that 70 to
78% of DEHP was biodegraded in an activated sludge test. Kurane et al.
(1980) showed that DBP, DEHP, and dimethy phthalate (DMP) were hydrolyzed to
the free phthalic acid in the presence of the bacteria Nocardia erythropolis.
DEP is present in the environment in small quantities compared to DBP
and DEHP. Background contamination from natufal.waters, reagents, solvents,
and other laboratory materials do not pose significant problems, thus making
it well suited for study in an aquatic ecosystem facility such as AEcoS.
DEP' is commonly used as a denaturant in the preparation of perfumes and
insect repellents with limited use as a plasticizer. Little information
. specific to DEP and its impact on the aquatic environment is available;
therefore, the aquatic fate of this compound has been inferred from data for
phthalate esters as a group (Versar, Inc. 1979). Based on such data, bioac-
cumulation and biodegradation are probably the most important biological
processes determining the fate of DEP in an aquatic ecosystem. Losses of
environmental pollutants can also occur through hydrolysis, volatilization,
photolysis, and sorption. However~ using the method of Radding et al.
(1977), we estimated from the half-life for alkaline hydrolysis that hydro-
lysis is not an important process. Volatilization of DEP as a transport
process can also be considered inconsequential since the vapor pressure of
DEP is extremely low (0.05 Torr at 700C). Only limited amounts of data
exist on the hydrolysis, photolysis, and sorption of DEP, and thus it was
important to investigate the effects of these processes on the fate of DEP.
The specific objective of the toxicant testing phase of this study was
to determine the fate of DEP in a complex aquatic microcosm. -Processes of
particular interest were sorption, volatilization, photolysis, hydrolysis,
and microbial degradation. Thus it was necessary to set up the experimenta-
tion such that the rate coefficients of these environmental processes could
be determined. Testing was conducted in two compartments to ascertain rates
of sorption, vOlatilization, hydrolysis, and photolysis; microbial degrada-
tion was studied under various environmental conditions in the other com-
partments. .
ASSUMPTIONS OF EXAMS and AEcoS
Implicit in the EXAMS parameterization experiments are several assump-
tions pertaining both to the model and to the experimental facility:
1. The chemical, physical, and biological processes that influence the
environmental fate of a compound are at steady state during the time inter-
val under study. Physical conditions (temperature, light intensity, turbu-
lence, etc) and the chemical regime (nutrient, toxicant, and water inputs)
in AEcoS remained relativ~ly constant under laboratory control throughout
the experiment, except during occasional mechanical failures. However,
temporal variability did occur in the biotic communities and in biotically
mediated processes. Therefore~ the validity of the steady-state assumption
was examined with respect to uptake and release of nutrients~ diel cycles of
metabolic gases~ and organic structure of the biotic communities.
2
.- -~ . '-~.-"T'7-'-
-------
2. Manipulation of chemical inputs into various compartments within
AEcoS creates a set of different environments in which to study the environ-
mental fate of a compound. To this end, different levels of inorganic and
organic nutrient enrichment were applied. Chemical and biological data were
analyzed to determine whether the chemical manipulations produced distinct
chemical environments and microbial assemblages.
3. Identically treated compartments in AEcoS behave as experimental
replicates. Chemical and biological data from duplicate AEcoS channels were
statistically analyzed to determine whether similar experimental treatments
resulted in replicable behavior.
3
" -. . ..-... - '-, ''''':-'~''r-- -- ,...."" - . .
~- -., -,-- -~ .-. -_. '-""--J -.. - '==-.--. -,... - '"
-------
SECTION 2
CONCLUSIONS
1. Sorption, volatilization, photolysis, and chemical hydrolysis were
insignificant processes in the fate of DEP. Microbial degradation of DEP
resulted in the disappearance of 36 to 90% of the compound. Although auf~
wuchs bacterial numbers varied by several orders of magnitude among the
microcosms, first~order degradation rates were similar in all except those
receiving low inorganic and high organic nutrient inputs. Therefore, only a
fraction of the total bacteria present may have participated in DEP degrada~
tion.
2. Aufwuchs communities appeared to reach a definable steady state
within two to three months of inoculation, based on available data for taxoe
nomic and non~taxonomic community structure (relative abundance of algal
species, similarity indices, chlorophyll a, ashefree dry weight, ATP, and
total organic carbon) and community metab;lic activity (relative changes in
dissolved oxygen and pH). Also, based on metabolic activity estimates, come
munities receiving direct nutrient inputs appeared to stabilize first, fol~
lowed by downstream communities.
3. A significant linear regression (r ~ 0.88) between phosphorus load~
ing and aufwuchs chlorophyll i suggested that phosphorus inputs were impore
tant in controlling non~taxonomic structure. A significant curvilinear
relationship (r = 0.91) between the same two parameters indicated that spa~
tial limitations may have become important in controlling aufwuchs develop--
ment at higher phosphorus loading rates.
Non~taxonomic parameters associated with living aufwuchs (chlorophyll a,
ATP, and bacterial numbers) provided greater resolution in distinguishing -
structurally distinct communities than did those that also included detritus
(ash~free dry weight and total organic carbon). As expected, microcosms
receiving nutrient enrichment displayed significant increases in non~taxono~
mic structure for all parameters examined. .
4. Replicate microcosms were generally similar with respect to both
chemical and biological variables. Nutrient concentrations (P04' N03'
NH3) were statistically indistinguishable for most of the experimental
period; the differences which did occur could not be attributed to experi~
mental manipulations. Non~taxonomic structure also was similar between
replicate microcosms. High light intensity, temperature, and nutrient load~
ing combined with low carbonate alkalinity appeared to select for euryecious
organisms typical of eutrophic systems. Algal species composition was
generally more similar than dissimilar (based on SlMI indices) among the
different microcosms. The differences which were observed usually resulted
from large variations in relative abundances of one or two algal species.
Shannon~Wiener species diversity and evenness estimates indicated relatively
complex algal aufwuchs communities.
4
-------
SECTION 3
RECOMMENDATIONS
1. In this study, DEP degradation was observed under environmental
conditions typical of eutrophic ecosystems. Further research is needed to
characterize the fate of DEP and its metabolites over a broader spectrum of
environmental conditions fu~d trophic states. .
2. DEP concentrations fu~d subsequent metabolites were assumed to have
no antagonistic or synergistic effects on the biologically mediated degrada-
tion of the compound. To verify this assumption, future experimental
designs should be expanded to include a range of chemical concentrations
permitting a simultaneous investigation of compound fate and effects.
s
-------
SECTION 4
FACILITY
DESCRIPTION AND EXPERIMENTAL DESIGN
Physical-Setup
This study was conducted in the EPA's Aquatic Ecosystem simulator
(AEcoS), a controlled-environment chamber that houses a 19.5 m long, 46 cm
wide, 51 em deep .U-shaped" flume. The Plexiglas flume is lined with
adhesive-backed 4 mil Teflon film. The flume is divided into two indepen-
dent channels (designated A and B), each subdivided into eight 250-liter .
compartments called Continuously Stirred Tank Reactors (CSTR). Each CSTR is
equipped with an outlet weir such that the effluent from an upstream CSTR
constitutes the influent for the next downstream CSTR. Uniform mixing in
each CSTR is accomplished by the use of Teflon-covered Plexiglas paddle-
wheels suspended longitudinally down the channel and adjusted to desired
mixing rates by variable speed controllers. The paddlewheel speed for this
experiment was set at 2.0 rpm.
The environmental chamber provides control of air and water temperature,
water flow, relative humidity, and light quality and quantity. The water
supply is well water treated by reverse osmosis and deionization followed by
ultraviolet sterilization, and flow was maintained at 500 liters per day at
200c during the experiment. The chamber temperature was maintained at
21°C with a relative humidity of 50%. The channel was illuminated with
2000 foot-candles of fluorescent light on a 12 hour light/12 hour dark cycle
using a radiant energy system located 0.6 meter above the water surface.
Nutrient chemicals were introduced continuously to selected CSTRs using
feed lines of silicone tubing and Harvard peristaltic pumps.
A concentrated solution of the toxicant, DEP, was added using an all
glass constant-head system (Figure 1); the metering system was set to 2.0
ml/mfn to achieve the desired final concentration when diluted with the
incoming water.
t~~~-thata~t~tistits
Figure 2 shows the experimental conditions of each channel with Channels
A and B treated as replicates. The purpose of this design was to estimate
the rate of degradation of a toxicant in different environments so each CSTR
was designed to simulate different conditions. Concentrations of elements
after final dilution in AEcoS are shown in Table 1. The pH adjustments will
be discussed in a later section. The purpose of each treatment is summa-
rized in the following sections.
CSTR I--
Neutral photolysis of DEP. This CSTR
nutrient-free water (345 ml/min) adjusted
input of 0.003 & NaOH (4.0 ml/min). CSTR
received a continuous input of
to a pH of 7.0 with a continuous
1 was not inoculated with biotic
6
-------
material and was continuously illuminated with two banks of black-light
fluorescent lamps to induce photolysis.
CSTR 2--
Alkaline hydrolysis
plus a solution of 0.03
also was not inoculated
of DE? This CSTR received the effluent from eSTR 1
~ NaOH (4.0 ml/min) to bring the pH to 10.0. eSTR 2
with biotic material.
eSTR 3--
Effects of -low. nutrient concentrations on community activity. (CSTRs
3-8 were inoculated weekly as described below.) This CSTR received the
effluent of eSTR 2 and the input of inorganic nutrient at a flow rate of 4.0
ml/min (Table 2). This eSTR also received an input of Hel to neutralize the
base added to eSTR 2. The concentrates were modifications of Freeman's
Reference Water (Freeman 1953) with the addition of micronutrients (Miller
et ale 1978) and 10% of the total amounts of Nand P added to AEcoS (0.062 g
NH4N03 and 0.011 g Na2HP04 . 7 H20 per liter concentrate).
eSTR 4--
Effect of sediment and low nutrients on DEP degradation. eSTR 4
received only the effluent of eSTR 3. This eSTR contained a stainless steel
sediment tray containing 100% unsterilized river sediments (Oconee River,
North Georgia).
CSTR 5--
Enhanced microbial degradation of DEP. CSTR 5 received the effluent of
CSTR 4 plus 4.0 ml/min of a concentrate of glycerin to stimulate hetero-
trophic growth. The glycerin was added such that a final concentration of
3.9 mg/l carbon was obtained.
CSTR 6--
Comparison of community development rate
received the effluent of CSTR 5 and an input
amount added to AEcoS). The concentrate was
and contained per liter: 0.558 g NH4N03 and
H20.
with eSTR 3. This eSTR
of high Nand P (90% of total
added at the rate of 4.0 ml/min
0.101 g Na2HP04 . 7
CSTR 7--
Effects of high amounts of Nand P on the rates of community develop-
ment. This CSTR received only the effluent of eSTR 6.
CSTR 8--
The influence of a natural sediment and high
of DEP. This CSTR received the effluent of CSTR
tray of 100% river sediments (same as eSTR 4).
nutrient regime on the fate
7 and contained a sediment
Inoculation
Biological communities were established in eSTRs 3-8 by inoculating each
eSTR with a I-liter sample from each of two local ponds and a creek. Each
replicate eSTR was then cross inoculated with a I-liter sample from the
7
-------
corresponding CSTR. Reinoculation and cross inoculation were performed
weekly throughout the experiment.
pH Adjustment
Experimentation was initiated on November 13, 1979. Nutrient inputs
were set up as described in the previous section and weekly inoculations
began. There was no addition of DEP from November 13, 1979, to May 23,
1980, to allow time for communities to become established. The addition of
Na salts to CSTR 3 during this period of time required the addition of 0.252
meq/l HCl for neutralization. DEP addition began on May 23, 1980. At this
time it was necessary to set up pH adjustments for CSTRs 1, 2, and 3. CSTR
1 required the addition of NaOH at a final concentration of 0.034 meq/l to
adjust the inflow water pH from 5.8 to 7.0. CSTR 2 required an input of
NaOH such that a final concentration of 0.358 meq/l NaOH was maintained to
adjust the pH to 10. This required an additional input of 0.358 meq/l HCl
to CSTR 3 to neutralize the base added to CSTR 2. In an attempt to lower
the overall pH in CSTRs 3-8, an excess of 0.358 meq/l HCl (final concentra-
tion) was added to CSTR 3. This excess acid was omitted after a period of 11
weeks in Channel Band 13 weeks in Channel A.
DEP"Experimental-Design
Initial investigation of the degradation of DEP showed 30-80% losses in
any individual CSTR containing a biological community. This made it impos-
sible tc study the degradation of DEPin more than one pair of replicate
CSTRs (e.g., A3 and B3) at a time. For this part of the study toxicant
input was sequentially moved downstream from one CSTR to the next on a
weekly schedule.
An aqueous solution containing 34.3 mg/l DEP was dripped in at a rate of
2.0 ml/min; analysis of the final dilution gave a mean input concentration
of 194 ug/l DEP.
After this part of the DEP study was completed, a separate study was
initiated to determine the sorption, volatilization, photolysis, and hydro-
lysis of DEP in CSTRs land 2 (in the absence of biological communities).
For sorption and volatilization all CSTRs were filled with nutrient-free
water. A continuous DEP input of 34.3 mg/l was added at a rate of 2.0
ml/min to CSTR I to achieve a final DEP concentration of 200 ug/l; analysis
of this final dilution gave a mean input concentration of 191 ug/l DEP. The
effluent from each.CSTR was the influent for the next downstream CSTR.
Samples from CSTRs 1-4 were analyzed in duplicate for five days to determine
the amount of volatilization and/or sorption of DEP to the surfaces of each
CSTR.
In order to determine the rates of photolysis and hydrolysis, pH adjust-
ments were set up as previously described for CSTRs 1 and 2 and the black
light fluorescent lamps were used. The input level of DEP into CSTR 1
remained 191 ug/l. Samples were analysed in duplicate for five days to
determine the rates of photolysis in CSTR 1 and rates of hydrolysis in CSTR
2.
8
-------
SECTION 5
MATERIALS AND METHODS
SAMPLING
Chemical
From November 20, 1979 to July 18, 1980, chemical measurements were
routinely performed in each CSTR four times per week: Tuesday and Friday at
0800 hrs (after 11 hours of dark) and Monday and Thursday at 1500 hrs (after
6 hours of light). Beginning with the July 22 additions of DEP, chemical
samples were taken in duplicate each Tuesday at 0800 hrs and at 1500 hrs.
Table 4 gives a summary of major sampling dates for the entire DEP experi-
ment. From August 5 to October 14 all chemical parameters were measured in
triplicate for the designated CSTRs. Oxygen and pH were measured in situ.
Water samples for toxicant analyses were pipetted directly from designated
CSTRs and extracted immediately in culture tubes with iso-octane. For other
chemical parameters (nitrite, nitrate~ ammonia, dissolved Kjeldahl nitrogen,
total dissolved phosphorus, orthophosphorus, and organic carbon), water was
collected with a glass tube, filtered through Gelman HT-450 filters (0.45 urn
pore size), and dispensed for analyses.
Biological
Aufwuchs samples were collected in triplicate beginning August 5, 1980,
using Teflon-covered plastic strips that had been colonized in AEccS for
four weeks prior to sampling. Measured areas of each strip were scraped,
brought to exact volume with distilled water, and blended for 5 seconds;
aliquots of the resultant slurry were dispensed for algal identification/
quantification, total microbial ATP concentration, chlorophyll i~ content,
ash-free dry weight, toxicant concentration, bacterial enumeration via plate
counts, and total organic carbon. ,"
Suspended samples were collected in
glass column. Samples were blended for
pensed for chlorophyll a content, total
bacterial enumeration via plate counts.
triplicate using a 38.0-mm (I.D.)
5 seconds and aliquots were dis~
microbial ATP concentration, and
The sediment communities in CSTRs 4 and 8 were sampled in triplicate by
collecting shallow (0.5 em) and deep (2.5 em) wire baskets contained in the
sediment trays. The surface algal mat was removed from the collected sedi-
ment samples, the material from the serliment baskets was mixed with a stain-
less-steel spatula, and aliquots dispensed for analysis of chlorophyll ~
concentration, total microbial ATP concentration, bacterial enumeration via
plate counts, and toxicant concentration.
CHEMICAL ANALYSES
DEI>
Concentrations of DEP in the water phase were determined according to
9
+.,-,- -- ...~.. -.,d, .-..~- --'-. .,..- .' ."-4.,-". .
-------
methods used in earlier studies with malathion (Paris et ale 1975) and
methyl parathion (Holm et ale 1981). Analyses were done on a Tracor 550
gas-liquid chromatograph equipped with a Ni-63 electron capture detector and
a 2 m (4 rom i.d.) glass column packed with 3% SE-30 on 80/100 mesh Gas-Chram
Q. Concentrations of DEP in aufwuchs and sediment samples were determined
on a Tracor MT-222Q gas-liquid chromatograph with a Hall detector using a
method described by Holm et al. (1981) for methyl parathion. Samples of
aufwuchs were scraped from Teflon strips and extracted directly in methy-
lenechloride with a Brinkman Polytron blender. DEP was extracted from the
sediments using a soxlet extraction method (Payne and Benner 1981). The
samples were initially extracted with acetonitrile and diluted to volume in
. hexane for analysis.
phospnorus. and. Nitrogen
Ammonia nitrogen (NH3-N), nitrite nitrogen (N02-N), nitrate nitrogen
(N03-N), orthophosphorus (P04-P), total dissolved phosphorus (TDP), and
dissolved Kjeldahl nitrogen (DKN) were analyzed in sample filtrates using
.EPA methods (1979) and a Technicon AutoAnalyzer II system.
Carpon
Total organic carbon (TOC) and dissolved organic carbon (DOC) were
determined directly using a Beckman 915-3 Toe Analyzer. Th€ method was
based upon the organic carbon combustion-infrared method outlined in APBA
(1976). Samples for DOC were filtered through Gelman HT-450 filt2rs~ All
samples \"ere acidified with 50% aqu':=o\lS ECI and spar':1ed ,
-------
boiling tris buffer (pH 7.75, 0.05 M) (APHA 1976).
then analyzed as described above.
Sample extracts were
Results were expressed as ng/l for the suspended community samples, ng/g
dry weight for the sediment community samples, and ng/crn2 for the aufwuchs
samples.
Chlorophyll' a
Algal biomass of aufwuchs, suspended, and sediment communities was
determined by fluorometric measurement of chlorophyll ~ {chI ~)conte~t
corrected for phaeophytin a (Strickland and Parsons 1968). Samples were
extracted with acetone using a B. Braun cell homogenizer and analyzed using
a G.K. Turner and Associates (Model III) fluorometer (Holm et ale 1981).
Results were expressed as ug/ml for the suspended samples, ug/g dry
weight for the sediment samples, and ug/crn2 for the aufwuchs community.
Algal"Species"Counts
Algal enumeration and identification was based upon the inverted micro-
scope method of Utermohl as described by Lund et ale (1958). A Zeiss"
invertoscope D with brightfield and phase contrast capabilities was used for
observation and identification of algal species. Samples of the aufwuchs
slurry were fixed with a modified acid Lugol's solution (Holm et ale 1981),
diluted to a known volume, and settled onto a sedimentation slide for enu-
meration. Algal density was estimated from observation of 400 algal units
or more of the total sample and at least 100 cells of the most abundant spe-
cies. Results were expressed as algal units/crn2.
Bacterial"Plate"tounts
Total bacteria were measured using standard plate counts (APHA 1976).
Total"Organic"Carbbn
Before analysis, slurry samples from the aufwuchs community were blended
for one minute to resuspend the community. A sample of this suspension was
taken and diluted to a workable concentration. The sample was then acidi~
fied to pH < 2 with 50% aqueous HCl and sparged for 3-5 minutes with N2
gas. At the time of analysis samples were sonicated for 30 seconds to
assure a homogeneous mixture. Samples were analyzed as described in the
chemistry section.
NUMERICAL AND STATISTICAL ANALYSES
Algal"species"biversity'(Evenness"ana"Siffiilarity)
Aufwuchs algal species diversity (H') was determined from the Shannon-
Weiner equation (Taylor et ale 1979).
11
..'- .. .~. .-:,. :-;"':-- -.i;;~I~. - ',. ",.wr"" . ..--- ,- '--"",- ,-,,' '; -..- "'-,""-1--.'''',' -"".' --'''', ' .~.-.-,_..,..., "". -".......,..,..~-,-. '-..' ".'" _. ~,- "'<;-. '''''''''.,.-.......- ,""" ~ .....~_.- ...........-,..,..- '-"--""""". '.~.'., -
'-'-"-"',_......"",... ~-'~-"'-~" ~ - - --
-------
H' = -
S
r Pi log 2 Pi
i=l
(1)
where P is the proportion of the i-th taxon in the sample, estimated from
ni/N; ni is the number of algal units/cm2 of the i-th taxon; N is the
total number of algal units/cm2 in the sample; and S is the total number
of taxa in the sample.
Aufwuchs algal species evenness (J) was calculated from the ratio
H'/Hmax (Taylor et ale 1979) where Hmax is the diversity obtained if all
taxa in the sample are equally abundant and is estimated from 1092s (S is
the total number of taxa in the sample).
SIAI'Indices
< .. . '.
. . . .'.' ..~ .. . - .' -' . .'
'..' ," The community structure of selected 'aufwuchs algal samples was compared
~by means of a similarity index (SIMI). SIMI has a maximum value of 1.00 .
~'when algal assemblages are identical between two samples and a value of 0.00
:'\:;'when' assemblages are totally different in. composition. The formula is, as
follows (Sullivan 1975).
S
r P.. P.
. 1J 1n
;" .1:;:1" . .. . -. -, -. " ,
SIMI =
S
r
i=l
2
P. .
1J
S
r
i=l
(2)
P. 2
1n
where Pij and Pin are the average relative abundances of the i-th
species 1n the j-th and n~th communities and S is equal to the number of
. species. . .' . .
. Aufwucns 'Biomass 'Analysis
]
Data from five estimates of autotrophic and/or heterotrophic components
of the aufwuchs community collected during DEP testing (chlorophyll ~,
microbial ATP, ash~free dry weight(AFDW)~ total organic carbon (TOC), and
.total bacterial numbers) were used in the following statistical analyses.
1. Two sample Student t-tests evaluated CSTR replicability of Channel A
versus Channel B.
2. One way analysis of variance and Student~Newman-Keuls multiple range
test evaluated CSTR treatment differences using combined data from Channels
A and B. : .
3. A matrix of product~moment correlation coefficients using combined
data from Channels A and B.
12
': -..-:':o.~
. 'r=:==- --C!:::~~ - - ~.... -.:
. - "'~-~ .-.. "1''']~'''~ -...........--,..-~~ ,.. ~ - ~ - ....... -~ - ~- -,... r 0>
-- ~ ----- -
-------
4. The Mann~Whitney test, a nonparametric analogue of
Student t-test, was used to evaluate treatment differences
and CSTR 6 based on data collected during the entire year.
the two sample
between CSTR 3
5.
tation.
Confidence intervals (a = 0.05) were constructed for graphic presen-
Because standard deviations were proportional to the means, data were
transformed by X' = 10910 (x+l) before parametric analyses were per-
formed. See Zar (1974) for a complete explanation of test statistics used.
tnemical'Enviro~rnent'Analysis
Concentrations of DEP in the water and first-order degradation rate con-
stants were analyzed using the Statistical Analysis System (SAS) (Servi~e
1972), and treatment differences were summarized using Duncan's multiple
range test. Differences between mean input level of DEP and concentrations
found during the sorption, photolysis~ and hydrolysis experiments were eval-
uated with two sample student t-tests. Student t-tests were used to test
chemical replicability of Channels A and B by comparing mean concentrations
of N03-N~ NH3-N, and P04-P for each channel from selected time periods
during the sampling year. Differences between morning ahd midday nutrient
mean concentrations were also evaluated by Student t-tests after combining
data from Channels A and B.
13
'. -:-0"---",---.' .~.......-.," ',;:".-... .-~--.~........-~ 'f""" - -."""...---r""-'..-,.' ...~-_.. '-'~...-....'" ..,'-... ""'-''''-~''--''''''--~ --- -:..-.._-,....-_r- .-" - '." -.- --~,-...,....... r--u-" ... ..*.,'......-...--
-------
SECTION 6
RESULTS
DIETHYL PHTHALATE
Concentrations of DEP found in CSTRs 1-4 in the absence of microbial
growth and nutrient additions (including pH adjustments) are shown in Table
5. Results indicate that there was 00 significant loss of DE? resulting
either from sorption to channel surfaces or from volatilization. No detect-
able concentration of DEP was found in the aufwuchs community based on a
total of 60 samples extracted and analyzed by the method described. A total
of 36 sediment samples extracted and analyzed also showed no detectable con-
centrations of DEP indicating that after the addition of DEP to CSTRs con-
taining microbial communities, no sorption to the sediments or microorgan-
isms occurred. There was no measurable loss of DEP by photolysis in CSTR 1
while the loss of DEP from water as a result of chemical hydrolysis at pH 10
in CSTR 2 was 4-5% (Table 5). The percent hydrolyzed was calculated using
the output from CSTR 1 as the input to CSTR 2.
Concentrations of DEP in CSTRs containing microbial growth indicated
that rapid loss of DEP occurred due to microbial degradation (Table 6).
first-order rates of loss, kl, for all CSTRs were calculated as follows
(Lewis et al. 1981).
The
k = L!V-~- (F!V) -t
I c
(3 )
where L is the mass of DEP added to the CSrrR per time (ms/nr) 1 V is the
volume of the CSTR (liters), F is the flow rate of the C5TR (l/hr), and C is
the steady-state concentration (mg/l) of DEP determined experimentally in
the CSTR. Table 6 shows a summary of the rate constants calculate6 in this
manner. Samples were taken for analysis beginning on SepteIT'.ber 9! 1980 q
PHOSPHORUS CONCENTRATIONS
Orthophosphate
Figures 3-5 summarize the orthophosphate concentrations in CSTR 3, 4]
and 5 over the entire course of the DEP experiment.
CSTRs A3 and B3 both exhibited rapid initial uptake of phosphorus; seven
days after inoculation, P04-P concentrations had dropped from a mean of
0.017 mg/l to 0.008 mg/l in C8TR AJ and 0.002 mg/l in C3TH 33. Concentra-
tionsthen became relatively stable, fluctuating between 0.002 mg!l and
0.006 mg/l in both CSTRs. During March 1980 P04-P concentrations in both.
CSTRs increased and exhibited much wider fluctuations, particularly in CSTR
B3.
Between March and June, P04-P concentrations in CSTRs A3 and B3 (Fig-
ure 3) showed wide fluctuations, but became relatively stable by July. Con-
centrations in both CSTRs remained at or below 0.006 mg/l until the end of
14
-------
the experiment. Neither CSTR reacted noticab1y to the cessation of acid
additions in August or the readditions in October and no effects were
noticed from the additions of DEP.
The P04-P concentration graphs for CSTRs 4 and 5 (Figures
very similar to each other and to CSTR 3 (Figure 3). Notable
P04-P concentrations around late March and early June 1980 in
CSTR 5 were most likely due to the P04-P increases in CSTR 3.
4 and 5) are
increases in
CSTR 4 and
The P04-P concentration graphs for CSTRs 6, 7, and 8 (Figures 6-8)
reveal higher standing concentrations, as well as a wider range of fluctua-
tions than in CSTRs 3, 4, and 5. In CSTR 6, as in CSTR 3, initial utiliza-
tion of P04-P was rapid, with concentrations falling from a mean of around
0.130 mg/l to below 0.030 mg/l within 2 weeks of inoculation. Concentra-
tions fluctuated during the next two months but remained below 0.060 mg/l.
After January 20, 1980, concentrations increased slightly but remained below
input levels. The incr~asing trend continued during April and May. Ob-
served peaks in P04-P concentrations in CSTR 6 at the end of May may have
been due to acid additions. This transient response was particularly not-
able in CSTR A6. P04-P concentrations increased until early July when
concentrations in CSTR A6 dropped noticeably below those in B6. Concentra-
tions increased in both CSTRs during the last month (November 1980) of the
experiment. Again, there was no apparant response to later acid manipula-
tions or DEP treatments. P04-P concentrations in CSTRs 7 and 8 generally
showed temporal patterns similar to those in CSTR 6.
Jotal-tissolved-Pnosphorus
Total dissolved phosphorus (TDP) concentrations (Figures 9-14) include
orthophosphate, hydrolyzable, and dissolved organic phosphorus. The TDP
graphs for CSTRs 3-5 (Figures 9-11) demonstrate the same trends as the
P04-P graphs (Figures 3-5) discussed in the previous section. TDP concen-
tration values were similar to the P04-P concentration values reflecting
low dissolved organic phosphorus in these CSTRs.
TDP concentration graphs for CSTRs 6-8 (Figures 12-14) followed the same
trends as the P04-P concentration graphs (Figures 6-8). These high nutri-
ent CSTRs, however, contained a greater percentage (> 20%) of dissolved
organic and/or hydrolyzable phosphorus.
NITROGEN CONCENTRATIONS
Nitrate-Nitrogen
Figures 15~20 present N03-N concentrations over time in all CSTRs.
N03-N concentrations in CSTR 3 dropped rapidly fr9m the input value of
0.12 mg/1 to below detection limits at the start of the experiment (Figure
15)~ remained at this level for about two weeks, and then increased to
between 0.02-0.06 mg/l where they remained for more than half of the experi-
mental period. During the last 5 months of the experiment N03-N concen-
trations increased to an average of 0.09 mg/1 by December 1980. In CSTRs 4
and 5, N03-N concentrations fell below detection limits after inoculation,
15
. -~-- """~-""'---"""-'-"-""-'-""~ "\..- .-.- "'-"'0;0"," .,'....-- --,'"".,..-,,--,.,- on.
..- -- -----....... ...,. ..- -",---"'"'.......--..,,,-- -,,,---,,-,.._-.,. -.-.............- .-' -"''''--'-~-----
-------
but gradually increased during the experiment (Figures 16 and 17). By
December 1980, N03-N in CSTR 4 averaged 0.04 mg/l; except for transient
peaks, concentrations in CSTR 5 never exceeded 0.02 mg/l. In CSTR 6,
N03-N concentrations decreased from the input value of 1.12 mg/l over a
period of 3 months, reaching a minimum of 0.50 mg/l in February (Figure
18). Concentrations gradually increased over the next 4 months to approxi-
mately 1 mg/l and remained near that level for the rest of the experiment.
Channel A showed a variation from this pattern only during May 1980 when
acid additions began. N03-N concentrations in CSTRs 7 and 8 (Figures 19
and 20) followed the same pattern as in CSTR 6, decreasing to a minimum of
0.40 mg/l in February and then approaching input levels.
Nitrite-Nitrogen
Nitrite nitrogen was not introduced into AEcoS as a nutrient and conse-
quently any accumulation of N02-N was due to chemical or biological activ-
ity. This would usually be in the form of ammonia oxidation, nitrate reduc-
tion, or excretion by organisms (Mortonson and Brooks 1980). No accumula-
tion of nitrite was observed during the DEP experiment in the low nutrient
CSTRs 3-5.
~.
;
~
~
In CSTRs 6-8 nitrite accumulated to concentrations exceeding those
usually found in natural systems. Figure 21 shows that nitrite concentra-
tions in CSTRs 6 began to accumulate within 2 weeks and steadily increased
to a maximum of 0.40 mg/l N02-N in CSTR A6 and 0.46 mg/l in CSTR B6 in
March. Concentrations then declined until both CSTRs reached 0.02 mg/l in
May. In June CSTR A6 showed a sudden increase to 0~39 mg/l that then -
declined to low levels in early Junei no increase was observed in CSTR B6
during this period. For the remaining 5 months of the experiment, N02-N
levels in CSTRs A6 and B6 remained relatively low (0.05 mg/l).
Nitrite accumulation also was observed in CSTRs 7 and 8 (Figures 22 and
23). compared to CSTR 6, slightly higher levels of N02-N were reached in
CSTRs 7 and 8 during the mid-March maximum. CSTRs A7 and A8 behaved simi-
larly to CSTR A6 in June, and bothCSTRs maintained relatively low N02-N
levels for the remainder of the experiment.
. ,
,
.~
Ammonia-Nitrogen
In CSTR 3, NH3-N concentrations dropped rapidly from the input value
of 0.125 mg/l to between 0.010-0.040 mg/l and remained at this level for
almost the entire experimental period (Figure 24). Channel A showed a
variation from this pattern during late May, when there was a transient
increase in the standing- concentration to 0.45 mg/li levels returned to
previous low valUes (0.040 mg/l) within two weeks. Channel B showed
increased NH3-N concentrations during May and August, but returned to
previous levels by the end of each month. NH3~N values in CSTRs 4 and 5
dropped to below 0.040 mg/l and generally remained at this level throughout
the experiment (Figures 25 and 26), except for a temporary increase during
May in CSTR A4. .
!
,
I
I
i
!
I
1
f
I
!
;
16
-------
Input of NH3-N into CSTR 6 was 1.125 mg/l. Average concentrations in
the water were 0.5 mg/l (morning) and 0.2 mg/l (midday) for the first four
months of the experiment (Figure 27). In March, these values decreased to
below 0.300 mg/l (morning) and below 0.010 mg/l (midday), but after two
months, returned to previous levels and remained relatively stable for the
rest of the experiment. Channel A showed an increase in NH3-N concentra-
tions to a maximum of 1.10 mg/l during the end of May, but returned to pre-
vious levels by the next week. In CSTRs 7 and 8, NH3-N levels were lower
than in CSTR 6, but generally showed the same temporal patterns (Figures 28
and 29).
Diss6lved"Kjeldahl"Nitrogen
Dissolved Kjeldahl nitrogen (DKN) in CSTR 3 remained near 0.2 mg/l
throughout the experiment (Figure 30). Channel A showed an increase during
May, due to the previously mentioned increase in NH3-N. In CSTRs 4 and 5
(Figures 31 and 32), there was no obvious trend in the data; values remained
near 0.2 mg N/l.
Dissolved Kjeldahl nitrogen remained near 0.6-0.8 mg/l in CSTR 6 (Figure
33); Channel A also reflected the increase in NH3-N during May.
In CSTRs 7 and 8 (Figures 34 and 35), concentrations in "the water
high week-to-week variability, but generally remained between 0.3 and
rng/l. Channel A showed the same pattern of increase in May that was
observed in CSTR 6.
showed
0.9
CARBON CONCENTRATIONS
Diss6lved"Organic"carDon
Dissolved organic carbon (DOC) concentrations remained fairly constant
throughout the experiment at levels dependent on nutrient treatments (Fig-
ures 36-41). In CSTRs 3 and 4, DOC concentrations averaged about 1 mg/l
with peaks as high as 4 mg/l. In the last month of observation, concentra-
tions increased rapidly to about 6 ~g/l in the midday samples. CSTR 5
received a continuous input of 3.9 mg/l DOC (as glycerin) and concentrations
remained near this level throughout the experiment, with peaks up to 9.0
mg/l in May and August. Midday DOC values again increased during the last
mon th .
In CSTRs 6-8, DOC concentrations averaged about 2 mg/l and showed a
steady increase to about 3 mg/l during the last four months. From J~ly on
less carbon analyses were performed due to instrument break-down. As in
previous CSTRs, midday DOC concentrations rapidly increased to about 6 mg/l
during the. final month of the exper iment.
Total"organic'Carbon
Total organic carbon (TOC) concentrations (Figure 42-47) were consider-
ably more variable in all CSTRs than were DOC concentrations, probably due
to sampling errors and to non-homogeneous distribution of suspended parti-
17
- -P. -:".R"'.. .'~":'- ~~_.._.., '~-'.': ~.--~: '....,. "> P",--- "....'. --... ¥ ,p' ,-- -. .." ".
-.. -- :. -', " "'~', ""~'-''',,:-' ...~_.~ -"!'~-"'~' ~- .-.-..-. ,--- -, '''''- -..... - .
-------
cles. In eSTRs A3 and A4, TOe was about 2 mg/l for much of the experiment,
but with major peaks as high as 12 mg/l. eSTRs 33 and B4 showed wider vari-
ations, especially after March. TOe concentrations in eSTR 5 were higher (5
mg/l) due to the glycerin input, but the temporal patterns were somewhat
similar to those in eSTRs 3 and 4.
From the beginning of the experiment until around April, Toe concentra-
tions averaged about 2 mg/l in eSTR 6,followed by wide variations especial-
ly notable in A6. Toe levels were slightly higher (average approximately 3
mg/l) in eSTRs 7 and 8. Toe variability in all CSTRs accompanied develop-
ment of the aufwuchs communities and apparently resulted from sloughing of
particulate organic matter.
DISSOLVED OXYGEN
As is typical in aquatic ecosystems, dissolved oxygen (DO) concentra-
tions (Figures 48-53) showed daily cycles associated with photosynthetic and
respiratory activity in the eSTR communities. Following an initial peak to
about 15 mg/l in early December associated with a bloom of suspended algae
. (unpublished data), midday DO values remained relatively stable (10-13 mg/l)
until May in all eSTRs except 4 and 5. In the latter systems, midday DO.
gradually increased from about 9 mg/l to 13 mg/l until May, and then fluc-
tuated between 8 and 15 mg/l. Morning DO concentrations remained fairly
stable throughout the experiment in eSTRs 3, 4, and 6, gradually increased
in eSTR 5, and gradually decreased in CSTRs 7 and 8.
Spikes in both midday and morning DO concentrations in May and June were
due to extended periods of darkness and light, respectively, caused by
malfunctions in the light system. The low midday values at the end of May
coincided with the initiation of acid/base additions.
Hydrogen Ion Activity (pH)
In ~he absence of acid/base additions to the channels, temporal pH pat-
terns (Figures 54-59) were relatively stable and closely resembled those
described for dissolved oxygeno This is due to the opposite metabolic
behaviors of 02 and C02' pH being an inverse function of the latter. pH
generally ranged from as low as about 6.5 in the morning to as high as about
10.2 in the evening; this wide range is probably due to intense metabolic
activity coupled tQ the low alkalinity (25 mg/1 caC03) of the medium.
During the first period of acid/base addition (~la:l - ,1!..ugust) t temporal
pH patterns were obscured and replaced by approximately 7-day cycles of
increasing and decreasing pH. After acid/base additions were stopped in
August, diel pH patterns once again became similar to those of dissolved
oxygen. Similar results occurred during and following the second acid/base
additions in October and November.
Several spikes in the pH data were due to mechanical malfunction of the
lighting system during mid-January, eariy May, and mid-September.
18
. ...~--~_.....,.............. ~,.- -- """".'-'-...---'--".-- '--""""'-"~'..._-..~-" -...-...-.~- '--.- -.- ,-. -. ,-., - '-
-------
A more detailed evaluation of the acid perturbations is discussed by
Kollig and Hall (1982).
AUIwucns" Communities
At the time of DEP experimentation (July - November), temporal develop-
ment of the biotic community" had progressed to the point where profuse auf-
wuchs growth covered all colonizable surfaces in the channel microcosms.
The development of an aufwuchs dominated community in "mature" microcosms
has been noted in previous studies (Dudzik et ale 1979) and is primarily a
consequence of the high surface area to volume ratios typically found in
microcosms.
The approximate values and percentages of chlorophyll a, ATP, and bac-
terial numbers found in the sediment, suspended, and aufwu~hs assemblages of
CSTRs 4 and 8 are presented in Table 7. Due to the large contribution of
the aufwuchs assemblage to total system biomass and to the observation that
the species composition of the suspended and aufwuchs algal assemblages was
generally very similar in any given CSTR (unpublished data), the present
evaluation is limited to analysis of aufwuchs data.
The aufwuchs assemblage biomass and species composition data are sum-
marized in Figures 60-64 and Tables 8-16. In addition, trace metal analysis
of the aufwuchs from each of the CSTRs was done during the week of December
2, 1980. Results of these analyses are presented in Table 17. Evaluations
of the results are found in the Discussion section.
19
0.,- p.-~-'.._--.~-...- ~....~..-.--_.. ''''-''''''-'',",~-~'~---'.'-'--.-':--'''--'''''p--''''''''''- .. .... ... ----~...- -...... '-- ----.............-...-~_. n ~ .".-. -- -- - ---- --~.. -.
-------
\
SECTION 7
DISCUSSION
ENVIRONMENTAL FATE OF DEP
Results of this study indicate that sorption, volatilization, photoly-
sis, and chemical hydrolysis were insignificant processes in the fate of
DEP~ Student's t-tests conducted on data shown in Table 5 showed no dif-
ferences between the mean input level of DEP and the concentrations found in
the CSTRs for sorption/volatilization (a = Oe05). A similar evaluation of
the photolysis data showed no significant differences. Evaluation of the
data on alkaline hydrolysis of DEP at pH 10 showed only a slight effect.
However, since it has been shown that the half-life of DEP resulting from
alkaline hydrolysis is proportional to hydroxyl concentration (Wolfe et al.
1980), and since the pH in AEcoS ranged from 6.5 to 9.5, it can be concluded
that alkaline hydrolysis was an insignificant process. These results corro-
borate the conclusions of Versar, Inc. (1979).
Results indicate that microbial degradation of DEP was the dominant pro-
.. cess in AEcoS. Table 6 shows that DEP was degraded from 36-90% in CSTRs
. containing biological communities. It has been previously determined (Lewis
et al. 1981) that bacterial transformation is by far the most significant
process in determining the fate of DEP in aquatic ecosystems. Figure 65
shows the first-order rates of degradation, kIt for DEP in AEcoS. These
data indicate that the rates in all CSTRs were very similar with the excep-
tion of CSTR 5 (Duncans Multiple Range Test, a = 0.05) even though there
were significant differences in both chemical environments and aufwuchs bio-
mass levels (Tables 8 and 18). Figure 65 further demonstrates that all
rates were within an order of magnitude which was surprising considering the
large differences in the biological co~munities.
Although the concentrations of bacteria in the CSTRs varied by several
orders of magnitude (Figure 64), there is some indication that on1y a frac-
tion. of the total bacteria present participated in the degradation of DEP.
This conclusion has been supported in an earlier study by Lewis and Holm
(1981) for both methyl parathion and diethyl phthalate in AEcoS. It also
has been shown by Lewis et al. (1981) that the rates of transformation in
aufwuchs dominated systems can be predicted within an order of magnitude
using second order rate coefficients based on total bacterial plate counts
of batch cultures. Any differences between predicted and observed rate
values may be attributable to the fact that only a percentage of the total
bacteria participated in the biotransformation of DEP. The need exists for
further investigation of the role of bacteria in biodegradation in terms of
percent degraders and of the precision and accuracy with which first-order
rates can be predicted from second-order rates using bacterial plate counts.
ASSUMPTIONS OF EXAMS AND AEcoS
Evaluati6ns'6I'steady~state
The concept of ecological succession toward some climax or steady state
20
-------
biotic community has long been a subject of discussion in the ecological
literature. Despite numerous debates about the mechanisms, directionality
or deterministic nature of the process, it is frequently suggested that
under constant environmental conditions, ecological succession results in
biotic communities of relatively constant species composition over some
period of time. Furthermore, this constancy of community structure is pre-
sumably reflected in measures of overall community function, such as nutri-
ent retention and gross productivity.
Several qualitative models have been proposed that detail expected pat-
terns of change as ecosystems develop toward steady state (Margalef 1968,
Odum 1971, Vitousek 1977). Empirical tests of these predictions have been
attempted in ~atural ecosystems, but long successional time scales and vari-
able environmental conditions have precluded firm conclusions. Investiga-
tions in laboratory microcosms that approach steady state under controlled
environmental conditions, however, have lent support to a number of the
predictions (Gordon et ale 1969, Cooke 1977, Wilhm and Long 1969, Kurihara
1978).
Since the EXAMS model rests on the assumption
the AEcoS system is used to validate EXAMS, it is
whether AEcoS achieves an operationally definable
imental time frames.
of steady state, and since
important to determine
steady state within exper-
An attempt was made to make such a determination with data from the DEP
experiment. This consisted of visual and, where possible, statistical ana-
lysis of the chemical and biological data. Temporal patterns of nutrient
retention, dissolved metabolic gas concentrations, and organic structure
were considered over the entire experimental period.
Nutrient Retention
Ecosystems can be considered to be at steady state when total nutrient
output is equal to total nutrient input (Jordan and Kline 1972; Vitousek
1977). In the present study, total input concentrations were relatively
constant for all elements. System outputs were measured as standing dis-
solved concentrations times output volume of each CSTR. Since particulate
outputs were not measured, complete nutrient bUdgets could not be. calculated
from the data: interpretations are therefore tentative. However, output/
input ratios were calculated for dissolved P04-P, N03-N, and NH3-N to
provide an estimate of retention patterns for these chemical species.
Phosphate was rapidly and almost completely utilized by communities
under both low (CSTR 3, Figure 66) and high (CSTR 6, Figure 67) nutrient
regimes. The latter communities thus incorporated approximately ten times
as much P04-P as the former, as reflected in significantly higher biomass
levels in CSTR 6 (Table 8). The output/input ratio remained low for most of
the experiment in CSTR 3 but gradually increased toward unity near the end
of the experiment in CSTR 6. The increase in suspended particulate organic
matter mid~way through tQe experiment (Figures 42~47) suggests that P04-P
(as well as other dissolved nutrients) comprised only a portion of the total
output and, therefore, the total output/input ratio may have approached
21
- ". ,~-. -'-""-"" r--, - -- ...... "',' " -. -- .-.. ~
. .-~ ,_.....- -.' <"--'.. ,_. -_. -.. ......-~.... ..'......- ~ - .- -', '-~, '00 ~.-. ~ ..'"-~. ~._-~ .-' ~ -- ...
-- --- ---
-------
unity. Net system loss of P04-P (i.e., output> input) also is plausable
since the communities in some CSTRS showed signs of senescense (increased
concentrations of phaeo-pigD~nts and P/R < 1.0, unpublished data) late in
the experiment.
Initial uptake of N03-N (Figure 68) and NH3-N (Figure 70) in CSTR 3
was rapid, probably corresponding to a bloom of suspended algae. Outputs of
N03-N then gradually inc~eased through time, approaching input concentra-
tion by the end of the experiment in CSTR A3. NH3-N outputs remained low
for most of the time, however, possibly indicating net uptake of nitrogen,
preferentially in the form of NH3-N. CSTR 6 (Figures 69 and 71) demon-
strated slight retention of N03-N during the first half of the experiment
but continuous uptake of NH3-N for the entire period of observation (ex-
cept in response to acid/base additions); preferential use of NH3-N is
ag ain indica ted.
In the absence of particulate import and export estimates, it is not
possible to determine whether the communities achieved steady state in terms
of nutrient retention. In some cases (P04-P in CSTR 3 and NH3-N in
CSTRs 3 and 6), however, there were periods of time in which the rate of
nutrient uptake remained relatively constant, especially during the early
months of the experiment. If, as indicated in Figures 60~64, total commu-
nity biomass generally reached a maximum level after about 2 months of
development, then particulate 1055 may have become an important factor rela-
tively early, possibly resulting in nearly equal total output and input
11utrient concentrations.
Met~bbli~'A2£i~i~~
-
r.1easurements of dissol'led oxygen (DO') were made weekly a.t lights-on and
6 hours later at midday. Die1 oxygen curVes measur~d on several occasions
(unpublished data) sho';,'ed thai: after 6 hours of light, DO concentrations
reached about 75% of the daily maximum. Therefore, the difference between
midday and morning rD conc~ntrations provides a relative comparison of net
daytime productivity, uncorrected for diffusion and accrual.
In general, all CSTRs (Figures 48-53) except 5 showed an initial burst
in oxygen production associated with the early bloom of suspended algae.
After about a month, production rates declined somewhat and approached fair-
ly stable levels in CSTRs receiving nutrients either directly (CSTRs 3 and
6) or in large amounts from upstream (CSTRs 7 and 8). Low nutrient CSTRS 4
aDd 5 required longer periods of time to reach stable rates of oxygen pro-
ducti.:m. and never achiev;?d. rate-s as high as, the others. Additions of gly-
cerin had little appare0.t effect on oxygen metabolism in CSTR 5; heterotro-
ph ic activity was probably limited. by the low concentrOations of inorganic
nutrients reaching this community.
In
sh owed
solved
the absence of acid/base additions~ diel
trends at least qualitatively similar to
oxygen. °
pH values (Figures 54~59)
those described for dis-
22
-------
eSTRs 3 and 6 reached stable daytime production rates after about two
months of operation. Without measurements of corresponding rates of night-
time respiration, however, ecological steady state (P/R = 1.0)-cannot be
conclusively defined. Diel oxygen curves measured during July 1980 revealed
P/R > 1.0 in all eSTRs, and during November 1980 P/R < 1.0 in eSTR 6 (unpub-
lished data). In static microcosms P/R typically approaches unity within
30-60 days (Wilhm and Long 1969; Odum 1971), but flow through systems may
require longer periods of time (Hendrix et ale 1981).
Aurwucns " Biomass
A highly significant relationship between total dissolved phosphorus
input concentrations and aufwuchs chlorophyll a concentration was estab-
lished from data collected during DEP additions. This empirical relation-
ship suggests that the relatively stable input concentrations of inorganic
nutrients into any given eSTR were primary factors controlling maximum
development of aufwuchs biomass; Wilhm and Long (1969) demonstrated a simi-
lar phenomenon in static microcosms. The overall stability of aufwuchs bio-
mass in all eSTRs during the sampling year; as measured by chlorophyll ~,
ATP, AFDW, TOe, and bacterial numbers; is shown in Figures 60-64. .EarlyW
data points represent the mean values of samples collected on 1/8/80 and
1/22/80 (n = 4); wmid. data points from 5/27/80 (n = 2), and Wlate. data
points during DEP additions (8/5/80 - 9/16/80, n = 6). These three time
periods roughly correspond to 2, 6, and 9 months of system development.
With the exception of the WearlyW Toe data, the 95% confidence intervals
around the mean biomass estimates for any given eSTR during DEP additions
mostly include the mean values from the two earlier time periods; this
observation suggests that aufwuchs assemblages attained a relatively stable
biomass level approximately 2 months after initial inoculation and main-
tained this stability throughout the experiment. The rather wide confidence
intervals around these estimates of community structure are functions of the
inherent spatial variability in aufwuchs distribution and associated samp-
ling problems, most notably loss of material from sloughing. This high
variability becomes particularly problematic when rigorous statistical eval-
uation of changes in structural parameters is attempted. In a recent study
using artificial streams, Rodgers et ale (1979) concluded that measured
functional parameters (metabolic estimates) were consistantly less variable
and therefore more sensitive to statistical evaluation than measured struc-
tural parameters. Although the functional parameters measured in the pres-
ent study (changes in pH and 02) could not be adequately resolved into
quantitative metabolic estimates, it is apparent from the available data
that inclusion of metabolic estimates in future studies could greatly en-
hance the ability to define system level stability.
Aurwucns" Algal" Species" Composition
In an attempt to evaluate the temporal stability of algal aufwuchs
assemblages in individual eSTRS, the SIMI index was used to compare algal
samples collected on January 22, 1980 to samples collected during DEP addi-
tions. SIMI values were based first on all taxa present in the sample and
then on the green and bluegreen algal divisions separately (see Tuchman and
23
~ ,," ". ,.-. - '':-''' ~-"'.~.- --....' -'- .. ":-~'-'>""'.,""," ~"'~_.~~"'-~-""-'.......,.....-.' .......... .".''''''-'~ ~ "~"'--"""-""-'-'-
. ..-..---..-_...~~.~--~--..~~..-.....~ ",.---.-.. ~ ...''''-''-~' ... --.......... -. .. I~ ~--.... ....... -.. -.....-- ~------
-------
Blinn 1979 for a similar treatment of algal divisions). Results of this
comparison are presented in Table 9; calculated SIMI values 2:.0.50 are arbi-
trarily considered to reflect similar species assemblages whereas values <
0.50 indicate dissimilar assemblages. These results should be considered
tentative because many of these comparisons are based on only one sample.
Based on all observed taxa, CSTRs 3-5 maintained generally similar auf-
wuchs species assemblages while CSTRs 6 and 8 developed dissimilar assem-
blages, with an apparent trend of decreasing similarity proceeding down-
stream. It is noteworthy that the bluegreen algae maintained highly similar
taxa in all CSTRs except CSTR 8, while the green algae developed generally
dissimilar taxa in all CSTRs except CSTR 4. Further resolution of the auf-
wuchs algal assemblages from these two sampling periods is provided by
Tables 10 and 11, summarizing the relative abundance of observed algal spe-
cies comprising> 1.0% of the total relative density. From these tables it
is apparent that the temporal stabilities of CSTRs 3-5 are primarily depen-
dent upon the dominance of Pnormidium ~., a filamentous cyanophyte that had
an average relative abundance of 76.5% in CSTRs 3-5 on January 22, 1980 and
33.6% during the DEP additions. The strong dissimilarity between SIMI
values for the bluegreen taxa in CSTR 8 is primarily due to the marked '-
decrease (from 36% to 3.8%) in relative density of Pnormidium ~. and to the
large increase (from 1.0% to 54.3%) in throococcus dispersus, a coccoid cya-
nophyte. The generally dissimilar SIMI values observed for the green algal
taxa in CSTRs 6-8 are mainly due to a temporal change in dominant Scenedes-
mus species; on January 22, 1980 Scenedesmus dimorpnus comprised 72% of the
total relative density of green algal taxa observed in these CSTRs, while
during DEP additions Scenedesrnus bijuga accounted for 44.8%.
In general, the existing data suggest that the aufwuchs assemblages in "
AEcoS reached fairly stable levels of biomass, metabolic activitY, and simi-
lar species composition within 2 or 3 months after inoculation. This is-
most. apparent in the nutrient enriched CSTRs (3, 6, 7, and 8); CSTRs 4"and 5
may have taken longer because of the low concentrations of nutrients reach~
ing them. In addition; CSTRs 4, 5, 7, and 8 may have experienced a time lag
in development corresponding to the distance from initial nutrient input.
Due to the high levels of nutrient loading and light intensity influenc-
ing all of the CSTRs, space may have been the ultimate factor limiting
growth in the communities. With continual sloughing of attached growth, the
CSTRs may have operated as chemostats~ the rate of particulate wash-out
equalling new growth for perhaps most of the experiment, but exceeding new
growth near the end. "
THE NATURE OF DIFFERENT ENVIRONMENTS IN AEcoS
One objective of the experimental manipulations in the DEP experiment
was to create different chemical and biological environments for toxicant
exposure in AEcoS. This section considers (1) the differences among the
biotic communities in AEcoS, (2) the behavior and possible influences of
phosphorus and nitrogen on these differences, and (3) the combined influence
of nutrient inputs and biotic community structure on the appearance of unex-
pectedly high concentrations of :1itrite-nitrogen in AEcoS.
24
-- ..- _._-----~-~.- '----.. --. -~ --... ~ -- - -"'''-''' .... ~ - -~-
-------
Taxonomic. structure
Differences in aufwuchs algal species composition in CSTRs A3~A8 and
B3~B8 during DEP additions were evaluated with the SIMI index. S1MI values
were first calculated for all taxa observed in the sample and then for the
green and bluegreen algal divisions separately.
Results of these comparison~ are su~~arized in Table 8. To facilitate
interpretation, comparisons that resulted in SIMI values greater than 0.50
were grouped together in a format similar to that of a multiple range test;
these results are presented in Tables 9 and 10.
In general, comparisons of aufwuchs algal species composition within a
channel are more similar than dissimilar and reflect the inherent difficulty
in establishing and maintaining distinct algal assemblages under the regime
of physical and chemical parameters used in the study. The combination of
high light intensity, abundant inorganic nutrients, low buffering capacity,
and relatively high water temperature resulted in selection for euryecious
organisms typical of eutrophic systems. Furthermore~ dissimilar SIMI values
were usually attributable to large changes in relative abundance of one or
two taxa; for example~ in Channel B, CSTRs 3~6 were grouped separately from
CSTRs 7 and 8 based on all observed taxa primarily because of differences in
relative abundance of Phormidium ~. and throococcus dispersus (Table 11 and
12). It is likely that i~~erent spatial variability, sampling errors, and
insufficient sample replicates also contributed to observed differences~
Calculated values of HI diversity and J evenness during DEP additions
(Table 13) ranged, respectively, from 1.69 to 2.69 and from 0.47 to 0.63 in
Channel A and from 1.27 to 3.14 and from 0.32 to 0.74 in Channel B. Ale
though these indices are sensitive to the same problems discussed above,
they do indicate that these systems were representative of relatively corne
plex aquatic ecosystem. There was no apparent relationship between CSTR
treatments and calculated diversity and evenness indices.
Non~Taxonomic-structure
An evaluation of the existence of structurally distinct aufwuchs assem~
blages in CSTRs 3~8 dur ing DEP additions was made by per forming a. one~way
analysis of variance on chlorophyll a~ ATP, AFDW, TOC~ and bacterial data
and summarizing any significant diff;rences with the Newman Keuls multiple
range test. Results of the multiple range test are presented in Table 14.
In general, estimators of living aufwuchs material (chlorophyll ~, ATP,
and bacteria) provided greater resolution in distinguishing structurally
different assemblages than did those estimators that also include detritus
(AFDW and TOC). For example~ chlorophyll a and ATP concentrations were
significantly different in 4 out of 6 CSTRS while AFDW and TOC estimates
were not shown to be different in any individual CSTR. Although export of
particulate material was not measured in this study~ it is possible that the
non~significant differences observed in AFDW and TOC.estimates can be par~
tially explained by sporadic sloughing of aufwuchs material and subsequent
export downstream.
25
. ~...-~..---,~...~-----...-.--._"-,, .",,-,"-'.-.~-'-.. ..........-., "-".- ,...,.---.--..-- ,..-.----.-.-.. -''io>'.' -...-~ ""","""'-r-.'" ,....-....- ---,.,..~,.,
-------
Results also indicate that different nutrient regimes influenced th~
measured structural components of aufwuchs assemblages. CSTR 6, which
received high concentrations of inorganic nutrients, was significantly dif-
ferent from low nutrient CSTRs 3-5 for all variables tested. CSTR 5, which
received glycerin to stimulate heterotrophic activity, had unexpectedly low
bacterial numbers; low inorganic nutrient concentrations apparently limit~d
organic carbon utilization in this CSTR and subsequent downstream export
resulted in the significantly highest bacterial numbers in CSTR 6.
Chlorophyll a, ATP, AFDW, TOC, and bacterial numbers collected for CSTRs
3 and 6 on the 8-major sampling dates (Table 4) were compared using the
Mann-whitney test, a nonparametric analogue of the Student t-test. This
analysis tested the hypothesis that the high inorganic nutrient load into
CSTR 6 would result in consistently higher biomass estimates than in low
nutrient CSTR 3. Results indicate that for all biomass parameters tested,
significantly higher (p = 0.05) biomass was observed in CSTR 6. Although
not unexpected, these results substantiate the differences observed between
these two CSTRs during DEP additions and support the assumption that struc-
turally different biological communities existed in AEcoS.
Trace Metal-cornpositibn
Trace metal composition of the aufwuchs samples (Table 17) were within
the ranges found in studies on tropical marine algae (Sivalingham 1978),
stream aufwuchs (Friant and Koerner 1981), and lentic phytoplankton (Goldman
et ale 1972). There is a wide range of algal metal concentrations in the
above studies because it is generally believed that algae take up metals in
proportion to the ambient environmental concentrations (Friant and Koerner
1981, Wong 1980).
Concentrations of Cu, Zn, and Mg per gram dry weight were higher in aL:f-
wuchs samples taken from CSTRs 6-8 than from CSTRs 3-5 (TablE: 17), Hanga-
nese concentrations, however, exhibited a general decrease from CSTRs 3-5 to
CSTRs 6-8.
In soils the availability of Mn and Fe (and perhaps other trace metals)
depends on pH, redox potential, and organic content (Mayer and Gorham
1951). These factors are also important in aquatic systems (Hutchinson
1973) and may determine trace metal accumulation. According to Hutchinson
(1973), Fe is in excess of Mn in most aquatic plants but the ratio of Mn:Fe
varies, generally being below 1.0. The ratios in AEcoS aufwuchs Here 0.70
(s = 0.22) for CST~s 3-5 and 0.14 (s = 0.05) for CSTRs 6-8. .
Ash peIcent (Table 17) ranged from 6.13 to 22.8%, and- generally agrees
with data from Goldman et ale (1972).
Influence-of Nutrients
The importance of phosphorus in relation to aquatic ecosystem develop-
ment and trophic state has been examined in some detail (Sakamoto 1966,
Vollenweider 1968, Edmondson 1972, Dillon and Rigler 1974, Schindler 1977,
Smith and Shapiro 1981). Current theory holds phosphorus to be the single
26
-------
most important limiting nutrient in many freshwater systems, and dissolved
inorganic phosphorus (P04-P) is considered to be the most available form
for plant nutrition (Chiaudani and Vighi 1974, Wetzel 1975).
The relationship between phosphorus concentration and chlorophyll ~ con-
centration has been the subject of research in both lakes and laboratory
microcosms. It has been found that maximum spring or winter concentrations
of phosphorus in lakes are directly proportional to summer planktonic chlo-
rophyll a concentrations in a nlli~ber of temperate lakes (Sakamoto 1966, Lund
1970, Dillon and Rigler 1974, Hickman 1980). It also has been observed that
many lakes exhibit concentrations of chlorophyll ~, nitrogen, and carbon
proportional to phosphorus concentrations, based on mean annual values
(Schindler 1978). The proportional relationship of phosphorus to chloro-
phyll ~ has also been demonstrated in both closed microcosms and simple
flow-through ecosystems (Fraleigh 1978). This section examines the relative
importance of phosphorus inputs in relation to aufwuchs standing crop in
AEcoS.
Mean total dissolved phosphorus input concentrations and aufwuchs chlo-
rophyll a concentrations after 4 weeks in each CSTR (Table 19) were used to
deve~op the log-log linear regressions shown in Figure 72. Channels A and B
are presented separately to demonstrate the replicability of the duplicate
CSTRs. The linear regression for both channels combined is also presented
to give the overall relationship of phosphorus to chlorophyll ~ in AEcoS.
The linear regression equations are
1. Channel A: 10910(chl ~) = 0.973 10910 (p) + 2.512 (r = 0.87, P < 0.001)
2. Channel B: 10910(chl~) = 0.855 10910 (p) + 2.566 (r = 0.91, P < 0.001)
3. Channel A and B: 10910(chl~) = 0.905 10910 (p) + 2.548 (r = 0.88, p< 0.001)
Literature values for correlation coefficients of linear regressions of
total phosphorus versus chlorophyll a from planktonic investigations include
r = 0.67 (Hickman 1980), r = 0.86 (Schindler 1977), and r = 0.98 (Dillon and
Rigler 1974); r values from the present study similarly refle~t a strong
correlation.
In an effort to demonstrate an even greater correlation between phos-
phorus input and aufwuchs chlorophyll a concentrations, two factors that
might cause deviations from linearity in the relationship wer.e considered.
These were limitations of nitrogen and light to the community. Dillon and
.Rigler (1974) used the data of Sakamoto (1966) to demonstrate that the
greatest deviations from the regression line of phosphorus versus chloro-
phyll ~ occurred in lakes with low N:P ratios. The N:P input ratios in
AEcoS, based on total dissolved nitrogen and total dissolved phosphorus
concentrations, varied from 14.8 to 65.3 during the period of study, none
falling below the N:P ratio of 12 considered by Dillon and Rigler (1974) to
indicate nitrogen limitation. Hickman (1980) observed that, as lakes prog-
ress from oligotrophy to eutrophy, decreases in light penetration and eupho-
tic zone depth cause the standing crop of phytoplankton to approach its
theoreticalmaximurn as a result of self-shading. He suggested that a curvi-
27
-------
linear relationship between phosphorus and planktonic chlorophyll ~ is mo~e
ecologically appropriate than a linear relationship and derived the equation:
10910 (chI ~) = 3.27 10910 (10910[P]) + 0.342
(4 )
where r = 0.842, P < 0.001. Data from Channels;.. and 3 combined (Table 19)
were used to determine whether such a relationship 2xisted in AEcoS. The
following curvilinear equation was derived
109lO{chl ~) = 2.439 10910 (lo9lO[TDP])
+ 3.583
(5 )
where r = 0.91, P < 0.001 (Pigure 73). Therefore, the curvilinear relation-
ship (r = 0.91) shows a stronger correlation than the linear relationship (r
= 0.88).
In addition to self-shading, aufwuchs communiti~s are affected by spa-
tial limitations not found in planktonic communities. Plankton are usually
surrounded by available gases and nutrients as they change position in the
water column. In contrast, periphyton are limited to substrates where dif-
fusion of gases and nutrients is restricted; even if biomass continues to
accumulate, only the upper layer of the periphytic mat may remain photosyn-
thetically active (McIntire and Phinney 1964, Sumner and Fisher 1979). As
Figure 73 illustrates/ chlorophyll a concentrations began to level off
despite increasing total dissolved phosphorus inputs. Therefore, the curvi-
linear relationship aoes seem more ecolo:::;icall'[ ,:.::;;:)[opri.ate for these auf-
',.;uehs communities.
Highly significar,t c'crr:elations among auf'liuchs chlocophyll ~, ash-fr'ee
dry weight, AT?, and TOC (Table 16) suggest that similar relationships exist
between phosphorus input and othe~ aspects of community structure. Thus,
manipulation of phosphorus inputs appears to be .:111 effective means of creat-
ing biotically distinct environments in AEcoS, particularly with respect to
non-taxanomic structure.
In this respect, the relative importance of nitrogen appeared to be less
than that of phosphorus. Inorganic nitrogen (NB4N03) and phosphoius
(NaR P04 . 7 H?O) were introduced at N:P weight ratios of 14.76 - 14.82
(CSTR 3) and l6q77 - 16.93 (CSTR 6, including inputs from CSTR 5) (Table
20). These values fall within the range considered to be optimal for algal
growth (Sakamoto 1966/ Lund 1970, Schindler 1977, Rh~e 1978). Inorganic N:P
ratios in the outflo~ing water (Table 20, Figures 74 and 75) were generally
higher than input values in both CSTRs, suggesting that phosphorus was taken
up to a larger deg~Ae than was ni~rogen. Therefore, downstream CSTRs (espe-
cially CSTRs 7 and B) may have .ceceived an excess of nitrogen relative to
phospborua, but with no pcoportional increase in periphytic growth. The
absence of nitrogen fixing brue-green algae nom all CSTRs (Tables 10 and
11) also suggests that nitrogen was present in abundance throughout the
experiment. Thus, phosphorus availability probably controlled growth of the
communities.
28
-------
Nitrite~NitrogenAccumu1ationin AEcoS
The accumulation of dissolved nitrite-nitrogen in CSTRs 6-8 during the
first half of the DEP experiment (Figures 21-23) was unexpected because of
the absence of significant amounts of nitrite in most aerobic natural
systems. Nitrogen in the form of nitrite was not introduced into AEcoS and
was therefore the product of chemical or biological activity. Nitrite for-
mation can result from .(1) oxidation of ammonia, (2) reduction of nitrate,
and/or (3) extracellular release of nitrite by phytoplankton (Mortonson and
Brooks 1980).
Nitrification involves oxidation of ammonia to nitrite and then to ni-
trate by the bacteria Nltrosomonas and Nitrobacter, respectively, although
other autotrophic and neterotropn1c organisms also may participate (Painter
1970). This process usually proceeds quickly from ammonia to nitrate, but
inhibition of the oxidation of nitrite to nitrate can allow nitrite to accu-
mulate in the system. Anthonisen et ale (1976) found that Nltrobacter acti-
vity was inhibited by high concentrations of free ammonia (NH3) and free
nitrous acid (HN02) in water treatment systems. Temperature, pH, and con-
centrations of nitrite and total ammonia controlled the concentrations of
the two inhibitory agents, free ammonia affecting the organisms at high pH
and free nitrous acid at low pH. Although Anthonisen et ale (1976) worked
with much higher concentrations of nutrients than those in AEcoS, inhibitory
effects might also be expected at lower concentrations. The constant input
of ammonia coupled with low alkalinity and high pH observed in AEcoS during
the period of high nitrite concentrations (pH> 9 for p.m. samples) could
potentially supply free ammonia and thus inhibit Nitrooacter activity. Bre-
zonik (1973) notes that a well buffered medium is necessary to avoid high pH
and ammonia inhibition effects.
In denitrification, nitrate is reduced to nitrite and then to ammonia by
phytoplankton, macrophytes and some bacteria, such as Escherlcla coli, Azo-
tobacter, and Lactobacillus. Other bacteria, including Micrococcus, Pseudo-
monas, and Denitrobacil1us convert nitrate to nitrite, ammonia, nitrous
oxide, or dinitrogen (Painter 1970). Since denitrification proceeds at low
oxygen concentrations, such conditions in AEcoS during morning sampling
(Figure 51-53) could have encouraged this process. In addition, thick peri-
phyton mats could have provided anaerobic microzones suitable for denitrify-
ing organisms. Comparison of the nitrite and nitrate concentration graphs
of CSTRs 6-8 (Figures 18-23) reveals an inverse relationship between the two
chemical species. This indicates that nitrite accumulation could have been
the result of nitrate reduction.
Excretion of nitrite by phytoplankton has been studied extensively in
marine environments (Vaccaro and Ryther 1960, Hattori and Wada 1971, Vortur-
iez and Herbland 1978) and more recently in fresh water systems (Ohmori
1978, Mortonson and Brooks 1980). Ohmori (1978) correlated excretion of
nitrite by Oscillatorla rubescens with nitrate uptake during the initial
growth phase of cultures. Nitrite accumulation was not affected by ammonia
concentration or by the presence of bacteria. Nitrite excretion was appa-
rently due to initially high levels of nitrate-reduction activity. Later,
nitrite-reduction activity increased and the cells assimilated both nitrate
29
-------
and excess nitrite. Nitrite concentrations in Ohmori's cultures reached
maximum values in 45 days and declined to undetectable levels by 70 days.
These findings resemble the pattern of nitrite accumulation in CSTRs 6-8 and
might explain why nitrite accumulated significantly only during the first
half of the experiment. Although Q. ruoescens was not observed, the pres-
ence of other species of Oscillatoria was noted (Tables 10 and 11). Ohmori
and Hattori (1970) have also observed nitrite accumulation due to Anabaena
cylindrica.
Thus, three processes may have contributed to the accumulation of ni-
trite in CSTRs 6-8: (1) inhibition of nitrification, (2) stimulation of
denitrification, and (3) algal excretion of nitrite. These processes may
have been promoted by a combination of high light intensity, nutrient -
enrichment, and low alkalinity which resulted in frequent occurence of high
pH and low dissolved oxygen concentrations, and in the presence of poten-
tially nitrite-excreting algae. The absence of measurable concentrations of
nitrite under lower nutrient conditions and less extreme variations of pH
and DO (i.e., CSTRs 3-5) but the presence of similar algal assemblages sug-
gests that processes (1) and (2) may have been more important than process
(3) in CSTRs 6-8.
REPLICABILITY AND DIEL PATTERNS
Chemical-Data
In order to evaluate replicability of chemical variables between Chan-
nels A and B of AEcoS and to compare morning and midday sampling, the nutri-
ent data were divided into four time blocks.
a.
b.
November 27, 1980 to January 20, 1980 (early succession)
January 21, 1980 to May 20, 1980
May 21, 1980 to July 20, 1980 (acid/base addition)
July 21, 1980 to December 5, 1981 (DEP study)
c.
d.
Replicability of A and B was tested first by calculating means of A and
B for each time block and then calculating means for morning samples and
midday samples (lumping A and B) for each of the time blocks, finally per-
forming t-tests on the comparisons. The nutrients tested in this manner
were N03-N, NH3-N, and P04-P in CSTRs 3 and 6.
The results in Table 21 show that for most of the experimental period,
there were no differences in N03-N between A and B for either CSTR. The
only differences were found in the last time block in CSTR 6 between Chan-
nels A and B, but it is not possible to conclude if this difference might
have continued. There were two time blocks in CSTR 3 that showed signifi-
cant differences in N03-N concentrations between sampling times, but these
comparisons involve values near the detection limit of the instrumentation
and may not have reflected a real difference.
There were no significant differences in NH3-N levels in Channels A
and B for either CSTR over the entire experiment. However, there were sig-
nificant differences in NH3-N levels between the two sampling times:
30
-------
NH3-N was usually lower at midday in both CSTRs 3 and 6, probably due to
biotic uptake during the light cycle.
There were no significant differences in P04-P concentrations between
CSTRs 3A and 3B except in the third time block, while CSTR 6A and 6B showed
significant differences in the first and fourth time blocks. CSTR 3 morning
versus midday samples showed significant differences in P04-P concentra-
tions in the first two time blocks, while CSTR 6 morning versus midday sam-
ples showed significant differences in the first and forth time blocks.
In general, Channels A and B were indistinguishable with respect to
nutrient concentrations for most of the experiment. The few differences
that were observed were not attributable to experimenal manipulations,
except perhaps effects of acid addition on P04-P concentrations in CSTR
3. Relatively greater P04-P uptake early in community development in CSTR
6A and 6B and diverging N03-N concentrations in the same CSTRs at the end
of the experiment may reflect random variation. As expected, diel cycles of
nutrient uptake and release were apparent, especially for NH3-N. Toetz
(1976) observed diel periodicity in concentrations of N03-N and NH3-N in
the epilimmion of a reservoir.
Aufwucns'Biomass'and'Soecies'ComDosition
.
Estimates of aufwuchs chlorophyll ~, ATP, AFDW, TOC, and bacterial num-
bers from Channels A and B during DE? additions were compared using the
Student's t-test (Table 22). Results indicate that CSTRs receiving direct
inputs of inorganic nutrients (CSTRs 3 and 6) had the most consistent repli-
cability, while CSTR 8 had the poorest. Of the 5 parameters evaluated, AFDW
was the most replicable with no significant differences, and bacterial esti-
mates were least replicable with 5 significant differences.
Comparisons of aufwuchs algal species composition in Channels A and B
via the SIMI index indicate that bluegreen algae were generally similar in
any given CSTR comparison while green algae were generally dissimilar (Table
23). Bluegreen algal similarities are primarily due to the ubiquitous pres-
ence of Pnormtdium sp. and the occurrence of tnroococcus dispersus in CSTRS
7 and 8, while green algal dissimilarities in CSTRs 6-8 result from the high
average relative density (46.4%) of Scenedesmus bijuga in Channel A and the
absence of this organism in Channel B.
31
-------
SECTION 8
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~ .... - .. ~ ~--- ~ ~ ~, ; - ~ - -~-~
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sampl'2so
-.) "
37
-------
Figure 1.
6 liters
3 liters
Toxicant Feed System
38
- -,,-- .........~..--~.., - -- ~
Flask 1
Flask 2
(constant head)
Flask 3
-- ~---.--~,-, :::::::~~:::~=~~~ --::::::~~::-' =::'~ -~;::~~~O::~:::-;: -;::::~-~;::::----~ - -- - -- - --- - _4___- --- - -----
-------
,/
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ACID LOW HIGH I
OR I INORGANIC I I 10 mq/I I INORGANIC: : I
BASE: NUTRIENTS: : GLYCE~IN : NUTRIENTS I I I
I J I I I I I
I I I I I I I
I I I I I I I
~~ ~~ ~~ ~~ ~~ ~~ ~~
~
VI
\D
AUTOTROPHIC
PHOTOL YSIS HYDROLYSIS SYSTEM mg/I RIVER mg/I
pH = 7.0 NO;}-N 0.125 SEDIMENT NO;}-N 1.125
NH;)-N 0.125 NH;)-N 1.125
P 0.015 P 0.1.35
1 2 3 4 5 6 7
RIVER
SEDIMENT
j
1 .
8
CSTR
CSTR
Conditions
H20 Flow - 500 '/Oay
Retention Time - 12 Hr
Mixing Rate - 2 RPM
H20 Temp. - 20°C
Light - 2000 Foot Candles
(12 hr light7 12 hr
,
,
i'
,
,
,
dark)
. Figure 2.
Experimental Conditions of Each Channel
I!' ,
. , .
, .
". .
, , ,
, "
'. "
. I .., . 'I 'I'! ; :
-------
./
(
I
I
!
.020
.015
-'"""'
"
01
5.010
a..
I
...
o
a...005
CSTR A3
MIDDAY
MORNING
- --
,
II
I
I
.000
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
~
0 .020 CSTR 83 MIDDAY - - -
MORNING
.015
-'"""'
" ~
01
5.01'0 ", .
"" ,
\ 'I" "
a.. " I., , "
I ' I ' " 1/ 1
... ..' I \, I
0
£L.005
.000
Figure 3.
NOY
AUG
NOV
DEC
SEP
OCT
JAN
FEB
MAR
MAY
JUN
JUL
DEC
APR
Orthophosphate Concentrations in CSTRs A3 and B3
-------
, .
, .
l
"
.020
.015
--.
'-..
0)
5.010
a...
I
..
a
0....005
.000
~
......
.020
.015
.r--.
'-...
(J) ,
5.010
a...
I
..
a
0....005
.000
Figure 4.
'.:' ,
. ..;
'.
, J I ",1
,
""1';'+"":'
,':': ".;."
CSTR A4
MIDDAY
MORNING
- --
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
NOY
DEC
CSTR
84
MIDDAY
MORNING
- --
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
NOY
DEC
Orthophosphate Concentrations in CSTRs A4 and B4
-------
.000 r
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
oj::. .020 CSTR 85 MIDDAY - -- -
N MORNING
.015
.,--...
'-..
\:J'I
-S .010
CL
I
... I, l'
0
CL.OOS I, I'" I I
V\d\ f') \ I 1
I ~ I 1 'I
-', 1 ~ '
, 1 I
1 '_I -
'I
.000 r
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
.020
.015
.r--
'-..
\:J'I
~.010
CL
I
...
o
[L.OOS
Figure 5.
CSTR A5
MIDDAY
MORNING
- --
Orthophosphate Concentrations in CSTRs A5 and 85
-------
0.16 CSTR A6 MIDDAY - - - !!I
0.14 MORNING
0.12
'""'
ci10.10
~0.08
a..
I.. 0.06 I,. I"'
a
a.. 0.04 . " ,\ , '
, " \ / '
If' ,
0.02 I "
~
0.00
NOY DEC JAN. FEf3 MAR APR MAY JUN JUL AUG SEP OCT NaY DEC
+:0. 0.16 CSTR 86 MIDDAY - - -
(.N
0.14 MORNING
0.12
...-..
~0.10
~0.08
a..
I.. 0 . 0 6
0
a.. 0.04
0.02
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 6
Orthophosphate Concentrations in CSTRs A6 and B6
-------
0.16 CSTR A7 MIDDAY --"",* m
0.14 MORNING
0.12
".......
~0.10 .
I
50.08 I
I
I
a.. -'
1...0.06 I
I
a I
a.. 0.04 I
I
I
I
0.02 I
--"
0.00
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
~ 0.16 MIDDAY
~ CSTR 87 - - -
0.14 MORNING
0.12
".......
~0.1 0
50.08
a..
1...0.06
0
a.. 0.04
0.02
0.00
NOY DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
.. . _.. .. - . .
Figure 7. orthoph?sphate Concentrations in CSTRs A7 and 87
-------
0.16 CSTR AS MIDDAY - -- Bf
MORNING
0.14
0.12
,,-...
~0.10 I
I
E A J
J""
"""0.08 I
I
a.. I
1...0.06 J
I
a J
a.. 0.04 I
I
.,
",,~., "
0.02
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
.s::. 0.16 CSTR 88 MIDDAY - - -
U1 MORNING
0.14
0.12 ,
,,-... \
~0.10 \
\
50.08 \
a..
I.. 0.06
a
a.. 0.04
0.02
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 8.
Orthophosphate Concentrations in CSTRs A8 and 88.. .
-------
.030 CSTR A4 MIDDAY - - -
MORNING
.025
-0.020
'-...
0'1
~.015
a..
~.010
.005 f)'
J
.000
NOV DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
.j:>. .030 CSTR 84 MIDDAY - - _.
C]\
MORNING
.025
-0.020
"
0'1
E .
'J.015
a..
~.010
.005
.000
NOY DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 9.
Total Dissolved Phosphorus Concentrations in CSTRs A3 and 83
-------
.030 CSTR A3 MIDDAY - - -
MORNING
.025
0.020
'-....
en
5.015
a.
~.010
.005
.000
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
.J::>. .030 CSTR 83 MIDDAY - - -
-...] MORNING
.025
0.020
.....,
en
5.015
a..
~.010
.005
.000
NOY DEC JAN FEB MAR ,APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 10
Total Dissolved Phosphorus Concentrations in CSTRs A4 and 84
-------
.030 CSTR A5 . MIDDAY - - -
MORNING
.025
-;:::- . 0 2 0
'-.
en
5.015
a..
~.010
.005
.000
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
""" .030 CSTR 85 MIDDAY - - -
oc.
MORNING
.025
-;:::- . 0 2 0
'-.
en
5.015
a..
~.010
.005
.000
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 11. Total Dissolved Phosphorus Concentrations in CSTRs A5 and a5
-------
0.30 CSTR A6 MIDDAY - - -
MORNING
0.25
i -::::-0.20
: '-..
U1
'50.15
a..
~ 0.10
0.05
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
.j:> 0.30 CSTR 86 MIDDAY - - -
\D MORNING
0.25
-::::-0.20
'-..
en
50.15
!1..
~ 0.10
0.05
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
i
Figure 12. Total Dissolved Phosphorus Concentrations in CSTRs A6 and B6
-------
0.30 CSTR A7 MIDDAY - - -
MORNING
0.25
00.20
"
tJl
-.SO.15
a..
~ 0.10
0.05
0.00
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
V1 0.30 CSTR 87 MIDDAY - - -
p MORNING
0.25
00.20
"
tJl
E .
,-,0.15
a..
~ 0.10
0.05
0.00
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 13. Total Dissolved Phosphorus Concentrations in CSTRs A7 and B7
-------
0.30 CSTR A8 MIDDAY ---
MORNING
0.25
-:::::-0.20
"-
tj!
50.15
C1-
~ 0.1 0 /
,. /
I ~
"
"
0.05 "
/
~~ _oJ
J\ -~.... -
""""--<...
0.00 T I I ,--r- I ~
NOV DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
U1 0.30 CSTI-( 88 MIDDAY - - -
I-'
MORNING --
0.25
-:::::-0.20
"-
en
50.15
C1-
~ 0..1 0
0.05 ~
"' I J
0.00 ~- -..
-, r---,- I -,
NOV DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 14.
'rotal Dissolveo Phosphorus Concentrations in CSTRs A8 and B8
-------
0.16 CSTR A3 MIDDAY - - -
0.14 MORNING
0.12
.--,.
610.10
50.08
z
1..,0.06
0
zO.04
0.02
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
I.J1 0.16 CSTR 83 MIDDAY - - -
N
MORNING
0.14
0.12
.--,.
610.10
50.08
:z
11')0.06
o.
20.04
0.02
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 15. Nitrate~Nitrogen Concentrations in CSTRs A3 and 83
-------
0.16 CSrrR A4 MIDDAY - - -
MORNING --
0.14
0.12
.........
ci10.10
.50.08
z
1,~0.06
a
z 0.04
0.02
0.00
NOY DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
U1 0.16 CSTR 84 MIDDAY - - -
V,J MORNING
0.14
0.12
,....
;'0.10
50.08
z
11')0.06
a
z 0.04
0.02
0.00
NOY DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
! Figure 16. NitrateeNitrogen Concentrations in CSTRs A4 and B4
-------
0.16 CSTR A5 MIDDAY - - -
MORNING
0.14
0.12
"..-.,.
'ci0.10
--SO.OS
z.
II') 0.06
a
z 0.04
0.02
. .
0.00 \ I \
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Vl 0.16 .CSTR 85 MIDDAY - - -
oj:>. MORNING
0.14
0.12
"..-.,.
'ci0.10
50.08
z
II') 0.06
a
20.04
0.02
0.00
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 17. Nitrate~Nitrogen Concentrations in CSTRs AS and BS
-------
1.4 CSTR A6 MIDDAY - - -
I
MORNING ,\
1.2 ,\
I
-""""' 1.0
"
01 0.8
E
'-J
z 0.6
I
..,
0 0.4
z
0.2
0;0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
V1 1.4 CSTR 86 MIDDAY - - -
V1 MORNING
1.2
.,-,. 1.0
"
01 0.8
E
'-.J
:z 0.6
I
..,
0 0.4
:z
0.2
0.0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
. Figure 18. NitratecNitrogen Concentrations in CSTRs A6 and B6
-------
1.4 CST'R A7' MIDDAY - - -
MORNING
1 .2
".-.., 1.0
'-..
01 0.8
E
'--'
z 0.6 -
I
0')
a 0.4
z
0.2-
0.0
NaY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NaY DEC
!J1 1.4 CS11R 87 MIDDAY -,- -
0\ :MORNING
1.2 ~
".-.., 1.0
'-..
01 0.8
.f
'--' ...
... ... I
:z 0.6 ""...../ '\ r
1 '/'1
0') 1/
a 0.4 "
z \
0.2
0.0
NOV DEC ,JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 19. Nitrate~Nitrogen Concentrations in CS'l'Rs A 7 . and B7
-------
l.4 CSTR A8 MIDDAY - - -
MORNING
1.2
""' 1.0
'-..
CfI 0.8
E
'-"
z 0.6
I
...
a 0.4
z
0.2
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
(J1 1 .4 CSTR 88 MIDDAY - - -
-..,J
MORNING
1.2
""' 1.0
'-..
CfI 0.8
E
'-"
z 0.6
I
PI
a 0.4
z
0.2
0.0
NOY DEC JAN FEE MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
; Figure 20. NitrateeNitrogen Concentrations in CSTRs A8 and B8
-------
0.7 CSTR A6 MIDDAY - - -
MORNING
0.6
........ 0.5
'-..
rn 0.4
E
'--"
z 0.3
I
..
0 0.2
z
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
U1 0.7 CSTR 86 MIDDAY - - -
00
MORNING
0.6
........ 0.5
'-..
rn 0.4
E
'--"
z 0.3
I
..
0 0.2
z
0.1
0.0 --
I
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 21. NitriteeNitrogen Concentrations in CSTRs A6 and B6
-------
0.7 CSTR A7 MIDDAY - - -
MORNING
0.6
,,-.., 0.5
..........
Q'I 0.4
E
'-'
z 0.3
I
..
0 0.2
z
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
U1 0.7 CSTR 87 MIDDAY - - -
\D MORNING
0.6
,,-.., 0.5
..........
Q'I 0.4
E
'-'
z 0.3
I
..
0 0.2
z
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 22. Nitrite~Nitrogen Concentrations in CSTRs A7 and 87
-------
0.7' CSTR AS MIDDAY ---
MORNING
0.6
........ 0.5
"'-
01 0.4
E
'---'
'z 0.3
' J
'"
a 0.2
z
0.1
0.0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Q\ 0.7 CSTR 88 MIDDAY - --
0 MORNING
0.6
........ 0.5
"'-
01 0.4
E
........,
:z 0.3
I
..
a 0.2
z
0.1
0.0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
F'igure 23. Nitrite~Nitrogen Concentrations in CSTRs A8 and 88
-------
0.5 CSTR A3 MIIYDA Y - - -
MORNING
0.4
,.-...
'-. 0.3
01
E
'--"
z 0.2
I
'"
J:
z
0.1
0.0
NOY DEC JAN FEI3 MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
0\ 0.5 CSTR 83 MIDDAY - - -
.....
MORNING
0.4
,.-...
'-. 0.3
01
E
'--"
z 0.2
I
'"
J:
z
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 24.
AmmoniaeNitrogen Concentrations in CSTRs A3 and 83
-------
0.5 CSTR A4 MIDDAY - - -
MORNING
0.4
.........
'-.. 0.3
C1
E
'--"
z 0.2
I
..,
I
z
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
0\" 0.5 CSTR 84 MIDDAY - - -
N
MORNING
0.4
.........
'-.. 0.3
C1
E
'--"
z 0.2
1
..,
I
z
0.1
"
I ,
0.0 ----_J
r I I
NOY DEC JAN FEB MAR APR MAY JUL AUG SEP OCT NOY DEC
Figure 25.
Ammonia~Nitrogen Concentrations in CSTRs A4 and B4
-------
0.5 CSTR A5 MIDDAY - - -
MORNING
0.4
..........
......... 0.3
0'1
E
'--'
z 0.2
I
'"
::r:
z
0.1
0.0
NOV DEC JAN FEB. MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
0\ 0.5 MIDDAY - - -
(".I CSTR 85
MORNING
0.4
..........
......... 0.3
0'1
E
'--'
z 0.2
I
'"
::r:
z
0.1
0.0
NOV DEC JAN FEB MAR APR MAY JUN' JUL AUG SEP OCT NOV DEC
Figure 26.
AmmoniacNitrogen Concentrations in CSTRs A5 and B5
-------
1.2
1.0
........... 0.8
.........
01
E 0.6
'-J
z
I 0.4
'"
:r:
z
0.2
0.0
'" 1.2
.j:>,
1.0
........... 0.8
.........
01
E 0.6
'-J
z
I
'" 0.4
:r:
z
0.2
0.0
Figure 27.
CSTR A6
MIDDAY
MORNING
-- -
I,
... I'.;" - 1 \
'_J ....."" ""',-'.I'J""'" ~-'-
, _J \ '\
J ... \ '\1' "I
-... '''-\/ \J
1
\
I
I
I
, I
I I
I 1
I I
I I
I ,
I I ,
, ~\ , ,
I \ " /',
\ J -.I --
1/\ \ ~.J....-... ....---- ./
'\,....., , , -
J - ~
--,
---
NOY
MAY
DEC
DEC
JAN
FEB
MAR
APR
CSTR
MIDDAY
MORNING
86
-- -
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
NOY
" ,
I' - - J' /,
"\ - - ./" "" .... ..... .... I '\1"\
...,/ 'J'/-" ,-..... .....J
JUN
JUL
AUG
SEP
OCT
NOV
DEC
Ammonia~Nitrogen Concentrations in CSTRs A6 and a6
-------
1.2 CSTR A7 MIDDAY - - - ~
MORNING
1.0 -
'"' 0.8
'-.
01
E 0.6
"--'
z
I 0.4
..,
::r:
z
0.2 1-,
1\... 1,./ '
" IJ \/ ",I '~,I
,/_.J ,--~,J \, 1\
0.0 .....~ \....._--
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
C]\ 1.2 MIDDAY _. - --
VI CST'R 87
MORNING
1.0
'"' 0.8
'-.
01
E 0.6
"--'
z
I
.., 0.4
::r:
z
0.2 ,\
I, - 1 \
',I '"..,-./ --........."\- \
. \
0.0
NOY DEC JAI'-J FEB MAR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 28. Ammonia""Nitrogen Concentrations in CSTHs A7 and 87
-------
1.2 CSTR A8 MIDDAY - - -
MORNING
1.0
..--. 0.8
'-
0"'
E 0.6
'-"
z
I
,., 0.4
:c
z
0.2
0.0
NOY DEC JAN FEB. MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
0\ 1.2 MIDDAY
0\ CSTR 88 - - -
MORNING
1.0
..--. 0.8
'-
0'1
E 0.6
'-"
z
I
,., 0.4
:c
z
0.2
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 29. AmmoniaeNitrogen Concentrations in CSTRs A8 and B8
._-~._--------- .
-------
0.8 CSTI~ A3 MIDDAY -- - -
0.7 MORNING --
0.6
"....... 0.5
"
0"1
E 0.4
'-"
:z 0.3
~
0
0.2
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
'" 0.8 CSTR 83 MIDDAY - - -
-...J
0.7 MORNING
0.6
"....... 0.5
"
0"1
E 0~4
'-"
-
z 0.3
~
0
0.2
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 30.
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A3 and 83
-------
0.8 CSTR A4 MIDDAY ---
MORNING
0.7
0.6
........ 0.5
'-.
en
E 0.4
"--'
z 0.3
~
£:)
0.2
0.1
0.0
NOV DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
0\ 0.8 CSTR 64 MIDDAY - - -
ex:>
MORNING
0.7
0.6
........ 0.5
'-.
en
E 0.4
"--'
z 0.3
~
£:)
0.2
0.1
0.0
NOV DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 31. Dissolved Kje1dah1 Nitrogen Concentrations in CSTRs A4 and B4
-------
0.8 CSTR A5 MIDDAY - - -
0.7 MORNING
0.6 J
......... ,I
, 0.5 /I
II
01 1 I
E 0.4 1 1
"-.J 1 I ~
, 1 1
z 0.3 ' 1 ,I
~ 1 I' \
0 1 I' \
0.2 ' ,I \
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
(j\ 0.8 MIDDAY - - -
1.0 CSTR 85
MORNING
0.7
0.6
",.-.,. 0.5
,
0'1
E 0.4
"-.J
z 0.3
~
0
0.2
0.1
0.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 32. Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A5 and 85
-------
1.8 CSTR A6 MIDDAY - - -
MORNING
1.5
.,...... 1.2
"
en
E 0.9
.........
z
~ 0.6
CJ
0.3
0.0
NOV DEC ,JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
-...J 1.8 MIDDAY
a CSTR 86 - - -
MORNING
1.5
.,...... 1.2
"
en
E 0.9
.........
z
~ 0.6
CJ J" ",;""'--"""""""'---,J.I-"',
.1.1 ,...... \
0.3
0.0 r
NOY DEC JAN FES MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
J
Figure 33.
Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A6 and 86
-------
1.8 CSTR A7 MIDDAY - - -
MORNING
1 .5
......... 1.2
'-
Q'I
E 0.9
'-'
:z
~ 0.6
£:)
0.3
0.0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
-...J 1 .8 CSTR 87 MIDDAY - - -
I-'
MORNING
1.5
......... 1.2
'-
Q'I
E 0.9
'-'
:z
~ 0.6
£:)
0.3
0.0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 34. Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A7 and B7
-------
1.8 C'STR AS MIDDAY - - -
MORNING
1.5
'"' 1.2
'-
0'1
E 0.9 -
'-'
:z
~ 0.6
0
0.3
0.0
NOV DEC JAN FEB. MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
--..J 1.8 CSTR 88 MIDDAY -- -
N MORNING
1.5
..--. 1.2
'-
0'1
E 0~9
'-'
:z
~ 0.6
0
0.3
0.0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 35. Dissolved Kjeldahl Nitrogen Concentrations in CSTRs A8 and 88
-------
12 CSTR A3 MIDDAY ---
MORNING
10
........ 8
"
0'1
5 6
0
0 4
0
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG OCT NOY
"'-I 12l CSTR 83 MIDDAY - - -
tJ.I
MORNING
10
........ 8
"
0'1
E 6
'-'
0
0 4
0
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 36.
Dissolved Organic Carbon Concentrations in CSTRs A3 and 83
-------
12. CSTR A4 MIDDAY - - -
MORNING
10
..---. 8
"
m
E 6
'---'
u
0 4
0
2
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
-...J 12 CSTR 84 MIDDAY - - -
.j::>.
MORNING
10
.......... 8
"
m
E 6
'---'
J
U I
o 4 I
o I
I
I
I
2 J
,J
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 37. Dissolved Organic Carbon Concentrations in CSTRs A4 and 84
-------
12 CSTR A5 MIDDAY - - -
MORNING
10
-'"""' 8
'-..
0'1
E 6
'-'
(.)
a 4 -' ~
0 -'
"
I
I
2
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
-..J 12 CSTR 85 MIDDAY - - -
VI
MORNING
10
---- 8
'-.. J~
0'1
E J'
6 I '
'-' I' --,
, - - '----\
(.) , .r -
a 4 ,"
0
2
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
,
Figure 38. Dissolved Organic Carbon Concentrations in CSTRs A5 and 85
-------
12 CSTR A6 MIDDAY - - -
MORNING
10
"....... 8
""
en
E 6
'--"
u
a 4
C)
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
" 12 CSTR 86 MIDDAY - - -
(]\
MORNING
10
"....... 8
""
en
E 6
'--"
u
a 4
CI
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 39. Dissolved Organic Carbon Concentrations in CSTRs A6 and B6
-------
12 CSTR A7 MIDDAY - - -
MORNING
10
""""" 8
"
trI
E 6
~
0
0 4
1=1
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
" 12 CSTR 87 MIDDAY - - -
-...]
MORNING
10
""""" 8
"
trI I
E 6 I
I
~ I
.() J
I
0 4 I
CI I
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 40. Dissolved Organic Carbon Concentrations in CSTRs A7 'and 87
... ~.
-------
12 CSTR A8 MIDDAY - - -
MORNING
10
'"' 8
"
O'!
E 6
'-'
u
0 4
CI
2
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
"-I 12 CSTR 88 MIDDAY - - -
00
MORNING
10
'"' 8
"
O'!
E 6
'--'
u
0 4
CI
2
0
NOY FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 41. Dissolved Organic Carbon Concentrations in CSTRs AS and 88
-------
12 CSTR A3 MIDDAY - - -
MORNING
10
........... 8
""
01
E 6
'-'
u
0 4
I-
2
0
NOY DEC JAN .FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
-...] 12 CSrrR 83 MIDDAY - - -
1.0
Iv.! 0 RNIN
10
........... 8
""
01
E 6
'-'
u
0 4
I-
2
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AlJG SEP OCT NOV DEC
Figure 42.
Total Organic Carbon Concentrations in CSTRs A3 and 83
-------
12 CSTR A4 MIDDAY ml!t- - -
MORNING
10
8 '
.......... 1\
" II
OJ "
E "
6 I ,
'--' I , I
U I , ~
I , "
0 4 I , ,
I- I , I
, , ,
,,' I , ,
2 "II ,"
" , '
" , ~
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
(XI 12 CSTR 84 MIDDAY I!t-[!}- - *
0
MORNING
10
......... 8
"
OJ ~ ~ I " ~
E .6 '
'--' 'I ~, , 1
, \
U ','I ",' " 1
" , I', ,I, ' 1
o 4 I ,
I- ' , I I'. I', ' 1
I ',,\I .' , '~' , 1
, ',' ,I ,
2 ' ' l '
, \ I I
,I'
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 43. Total Organic Carbon Concentrations in CSTRs A4 and B4
-------
00
I-'
12
10
...--...
"
c:n
E
'-"
8
6
u
a
f--
4
2
o
12
10
-'"""'"
"
c:n
E
'-"
8
6
u
a
f--
4
~~
csrrR A5
MIDDAY (!) - - -!!H~W
M 0 RNIN G ----r--
'I
NOY
NOY
DEC
NOY
DEC
JAN
FES
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
NOY
,
I' 1
I ' ,
Ji\~/" M .'
j \ J \ j' J ,
I/"X--.-J. J 'J J :-. 0.. "-1 \,
'l" v , W \,<1\ :
y "
DEC
JAN
FES
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
CSTR
MIDDAY
MORNING
- --
Figure 44.
(!)
(!)
85
I,
.... -\ I \
,..,.""" \ I \
-- '"
" ,
" '
,
Total Organic Carbon Concentrations in CSTRs A5 and B5
DEC
-------
12 CSTR A6 MIDDAY .. I!J
MORNING
10 I
~
I'
.---.. 8 ,\
I'
" 1\
tJ1 I '
E 6 I '
.......... I \
0 . I '
I I
0 4 I \
I- I I
J 1
--- ,
2 I \
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV. DEC
oc 12 CSTR 86 MIDDAY * -- -
N MORNING
10
.---.. 8
" I
tJ1 I
E 6 I
'-' I
0 I
0 4 -'- -
I- ./ "\
\
\
2
0
NOV DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC
Figure 45. Total Organic Carbon Concentrations in CSTRs A6 and B6
-------
12 CSTR A7 MIDDAY - - - ~ *
MORNING
10
I
,
.......... 8 '
"- '
,
01 I
E 6 I
'--"
()
0 4 "'
I-
"' "'
"'
"' "'
2 "' "'
"'
"'
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
00 12 CSTR 87 MIDDAY - - -
(.N
MORNING
10
I
.......... 8 ,
"- 1
J' 1
01 "' \ '
E 6 " \ 1
'--" " \ 1
"
" \ I
() , \ 1
0 4 I \ 1
I- I \ I _I
----.....
... ....,"' ''
2
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 46. Total Organic Carbon Concentrations in CSTRs A7 and 87
-------
00
.j::.
12
10
........
..........
01
E
'---'
u
a
I-
12
10
........
..........
01
E
'---'
u
a
I-
Figure 47.
CSTR A8
MIDJ;AY
MORNING
I
'I
'I
1
I
,
,
- --
(!)
"
8
6
to 1\", I
,\ , .. I
,\ I I
I \ I
I \ ,
, \ I
I \I
,
...
.,.-.,"
"'"'
"'
"'"'"'
4
2
o
NOY
JAN
FEB
APR
MAY
JUN
JUL
SEP
OCT
NOY
DEC
AUG
MAR
DEC
CSTR
88
MIDDAY
MORNING
~
,I
"
,I
,I
"
I I
I I
- --
**
" , E*
I'
,
I ,
I ,
,
,
,
,
,
,
,
,
,
,
,
,
,
,
,
"
,-...
......
8
6
4
I
I
, I
, -
"
{
2
o
NOY
JAN
APR
MAY
JUN
AUG
SEP
OCT
DEC
JUL
FEB
MAR
DEC
Total Organic Carbon Concentrations in CSTRs A8 and B8
NOY
-------
18
15
"....... 12
"
0'1
E 9
"'--'
a
CI 6
3
o
00 18
U1
15
"....... 12
"
0'1
E 9
"'--'
a
CI 6
3
o
Figure 48.
1
I I
I I II '
I' t' h.. 'I I ,A...
", ,I I I 'I I \ J \
. 'II I, r'~ II ',,/1 I ~" ,I, I '" ",-'" \
, . .... I, J' I 1\ -' I I I' I I I I \ 1 J' \-
I ........ ""_J''''..I \ " \ ' " '",""""--"
I I I I ,
, ,-,
I I ... .
, I ' , J'
I ,'"
I,
"
I
I
CSTR A3
MIDDAY
MORNING
- --
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
CSTR
MIDDAY
MORNING
--
83
- --
~
\ I I
I' - I' J J \ 'I I' /, - 1 I .
II ~ " ,/1,,\/\
I I - ....-",\1 \1"'" '\/1111 \ II
I lA, I '...",...""J . "\ 11 \,\1 II 1111 \../ \
1 ,I . . 1 I , . J
1 '" '\. .....,
I II \ ,-
I II, 1
I II',
I 11
U
,
SEP
OCT
NOY
DEC
..
.. J ,
I '\ ,"'" "\ .",
I '\ J "'"""",,,
'I '",'"
NOY
AUG
DEC
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUL
Dissolved Oxygen Concentrations in A3 and B3
SEP
OCT
NOV
-------
18
15
..--... 12
"
rn
E 9
"-'
o
£:) 6
3
o
00 18
0\
15
..--... 12
"
rn
E 9
"-'
o
0 6
3
o
Figure 49.
J'
J '
J '
~ ' ~
. /,'", I'~,,,,,,,"-' I
_/,-' 'J', ~,.. ,'\ I
-, J, ,\.J" " " ' ,
J I, --,--' "'-' ~ ' I
\ J ~ .. / , I
~ ----' .J , I
, ,
"
II
II
,
CSTR A4
MIDDAY
MORNING
- ---
.
,
,
'_'\ ,-
, "
, ,/--- ,
,-,/ ,
-''\
--...._-- '.....,.,.....,
'\
'\ J
, J
..,
NaY
JAN
MAY
DEC
CSTR
84
FES
MAR
APR
JUN
JUL
AUG
SEP
OCT
DEC
MIDDAY
MORNING
NOY
- --
I
, r- n I
~" " ~ I '\ I ~ I' 1\. .
-' \, , '\ , , v' " ,I' ( , " '\
I .. "J" I , ,/,' , J""
-"', ./ \J " I I " "\' \ ./ ....
.1.1', ./,.1,\1 ~ - ,,' " ,I \ ./- J-' ,
./, , I,' -"' '.../
,-- ./ I, \ I
- " \/
"
./ ,
v ,
'--
NOY
DEC
JAN
APR
MAY
JUN
JUL
SEP
NOY
DEC
FES
MAR
AUG
OCT
Dissolved Oxygen Concentrations in A4 and 84
-------
18 CSTR A5 1vIIDDAY - - -
MORNING
15
...."'
12 I \
\ ---",""-"'"
, ..--.. ....,
'-.. '
i ....; ,
0'1 ,
E 9 ,--
~
0
0 6
3
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
00 18 85 MIDDAY - - -
...... CSTR
MORNING
15
..--.. 12
'-..
0'1
E 9
~
0
0 6
3
0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
: Figure 50. Dissolved Oxygen Concentrations in A5 and B5
-------
18
15
......... 12
""
rn
E 9
'-J
a
a 6
3
o
00 18
00
15
......... 12
""
rn
E 9
'-J
a
a 6
3
o
, Figure 51.
CSTR A6
MIDDAY
MORNING
- --
J,
1 , II
J , 1" J'" . 1 I \ "
" \ J, 1 1 J \ I' . J- \ I \ -... J'
\ - ,,,'-- . J ',IJ \... \ .. J\ - J \" \, -, '" ,
.... - - - - ,J .... - '" J , ~, J,'" I"'" J \1 .., \ ...- "\,
*"" ',I', J \...-- """....
\I ' I '.
II 1 I
1 I
I J
II
II
,I
II
,I
-
NOV
DEC
JAN
MAR
MAY
JUN
AUG
SEP
OCT
NOV
DEC
JUL
FEB
APR
CSTR
MIDDAY
MORNING
- --
86
I,
1 \
1 1
1 1
1 1 J\ - I
J \ .,1,.'/ \---\......., .,~ ~ /".... I \.,. ~ I i'"
\ I' - I \ -...' '''' \ I J \ I " II _I \ - -I ,'" -,
'I ... J - / 1/ \1" ...". - ...,
, I II \ '" \", "...
" '",'- 'J "',,," ,
'.
'.
I,
~
"'---
NOV
DEC
JAN
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
NOV
DEC
FEB
Dissolved Oxygen Concentrations in A6 and 86
-------
18
15
"....." 12
"
I:J)
E 9
'--'
a
0 6
3
o
00 18
to
15
"....." 12
"
I:J)
, E 9
'--'
a
0 6
3
o
. Figure 52.
MIDDAY
MORNING
---
CSTR A7
"
I ,
I ,
I , ,
, , I t, j'.... "
" ,." 'I '\ I
""./' ~- '"" J 'I' ,\1 \ , -
",,,'" "'-, .I" II I. 1-'
'-' J IJ II'.... I
IJ , I
1 '
,I
"
,I
II
,I
I'
1
I" .
1 '" "
I 'I' "
-- ,,' I I
\ , I I
'.,,'''-'"' I , ....---
" '...,""'~'" '--,
,
,
NOV
JAN
MAR
APR
JUN
SEP
NOV
DEC
JUL
AUG
OCT
MAY
DEC
FEB
CSTR
MIDDAY
MORNING
- --
87
,
I' ,"
\ ,
I , J' ",
I , II , ,
\ ,..- III \
ow" -./ \." ." " I ,I '"-
...." "" "-\I'
'.,/ -.,-'"\ / ..
~
"
I' j' ../\
,I , I "", ,,-' - , I"
," . , ... , --
, I "- ,,", "I .... - "
I I , I', "....
, , ,...J \
,I " ...
,I -...
,I
,I
11
I
I
~
NOV
JAN
MAR
APR
JUL
SEP
DEC
MAY
JUN
AUG
OCT
NOV
DEC
FEB
Dissolved Oxygen Concentrations in A7 and 87
-------
18
15
"..... 12
"
t!I
E 9
'--'
o
£:) 6
3
o
\D 18
o
15
"..... 12
"
~
E 9
'--'
o
£:) 6
3
o
Figure 53.
MIDDAY
MORNING
CSTRA8
"
, ,
- ,
,
, - 1"""''''- I A
, ,.-_J '\1 '--" 1\, I" ,- "
I J I '- , '" J .... - 'I " J ,
.. , 1 '" ", '
\ 11,,.1
. , 1
, 1
I 1
, ,
, 1
11
\I
II
.
.
---
/, I,
{\ ~ I,
,J , I , I, -,
1 ,I' 1 J,
"',' ',., ./'- ""'.J" \" '-.;,
,
...
,
,
NOY
~
MAY JUN
DEC
DEC
MAR
APR
JUL
SEP
JAN
FEB
AUG
OCT
NOY
MIDDAY
MORNING
CSTR 88
1,1,
, ,
, ,
I ,
',I, -,-,,,,--"'\, r1' J
, J , 1\' L I I
'- ... , 'I "', 1 \ .. 1'1 '
''''', ',' 'I -, 1 ," I" -
,J -oJ Ij '\ 1 'II \ ,,' J - ,-
I ..," , ." ~
I I'" " --
I I ,J.
, I
, I
I,
"
I,
I,
\
I
- --
--
" J'"'" '\
J '., .....-
- \
\
"
'---
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
NOY
DEC
Dissolved Oxygen Concentrations in A8 and a8
-------
11 .0
10.0
9.0
8.0
I 7.0
Q..
6.0
5.0
4.0
3.0
I.D
~ 11 .0
10.0
9.0
8.0
I 7.0
CL
6.0
5.0 -
4.0
3.0
Figure 54.
CSTR A3 EVENING
A': ._r~ MORNI~G
I , J ~ - .I., , ,. -., J I I J,"'" ....
J I - - - .., 1-"" , / J oJ" '""- \ "II 1
I It ~r" J .," Ii 11
" ' ,
'I II ,
\ II ,
,
\ I I
I ,
I
,
,
,
- --
, . J ..
, " II .' 11
,I "I,ll "
1"'111' t, I, 1
I , 1111" J I I, 1
, , , " \1 I , b I, ,
I I I " . I " 'I" I
I 'I ~ "I 'I I I
I I, I ,I II I
I " II '..'1 1 :
'J I I-
I
"..J~, "'--~_./\......
I " \
,I 1 I ~ '
I' ,1, . '
1'111 '
11,'1 '
,','I I
"III I
J ','I '
I , "
I \/1,
, '
----r-
APR MAY
.JUN
JUL
AUG
NOV
DEC
JAN
FEB
MAR
CSrT'R 83 EVENING
/~J'J'-"'\ _J -. ,... ~9~J!~/:t:Ig"r{ ,./ '\/-\J~ I"\J'~ ~
I '~J\' "h' 11 I I, ,
I, . , ,'I ,'. 1
A I( 'I I I I ,I ~ II & I
fll I, 11 ,'1111'11111 I
11\ ~ :', I I I II I: : I II II I 1 I
I \ \ l' I', I ,II II ~ 1 I
'~,..J!, 1,1,,1'111111,,1
v'~.~(iJ\A.y~ ,'~' ::::::::::\~:I :':
J , IM,III II \" . I',
I ~~~\ ":
- --
SEP
OCT
DEC
NOV
NOV
,---
DEC
,-
MAY
r--
AUG
,JUN
JUL
,JAN
FEB
MAR
APR
pH Values in CSTRs A3 and B3
SEP
OCT
NOV
I~
DEC
-------
\0
N
:r:
Q..
11.0
10.0
9.0
8.0 -
7.0
6.0
5.0
4.0
3.0
J:
a.
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
Figure 55.
CSTR A4 EVENING - --
, ,.... ¥OR:r~~lJijP..I\.."~"'-'-' -J"" ,~ , . " ~
oJ ....'...'\.- }16 -,/'" '""""\,",.1' ",.', 11"-1 1,',,'" 1 ..,...'" -J..,A'J~J~--.c "
..~... " I' I'" ~ \, ,I I 1\ .. I" I A , I....r
" , I 1 '" 1\ I 1\ , 11 I 1 I,
i I I I 'II ,I /1 In, U I I'
I I I I I I I I I ''', ,
I I ' 'I " I I ,II, I I
I I II I I' I I ~', I I
I , , I: I I ' " : '
I , I I , It 1
I I , I
I I I, II U\J"
, ' " II
I I, II
I I, I
I I
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUl
AUG
SEP
OCT
NOY
DEC
CSTR 84 EVENING
MORNING
"
~ l /I' ", 1 ,
r_~ J~ '-"\ ,-I"", ''''''.~' ,Iv t'./ "-11,
: .. I'V. 1/11 ,....' .
,
,
- --
NOY
DEC
JAN
FEB
MAR
APR
MAY
JUN
JUl
AUG
SEP
OCT
NOY
DEC
pH Values in CSTRs A4 and 84
-------
11.0 CSTR AS EVENING - - -
MORNING
10.0 ,-_-J"\/""I
9.0 - - ,- ...',tl' '1 I
'-'~A-- - -J . .
, I
8.0
:r: 7.0
a.
6.0
5.0
4.0
3.0 I
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
\D 11 .0 CSTR EVENING - - -
tN
10.0
9.0 r
8.0
:r: 7.0
a.
6.0
5.0
4.0
3.0
NOY DEC JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOY DEC
Figure 56. pH Values in CSTRs A5 and B5
-------
11.0
10.0
9.0
8.0
I 7.0
a.
6.0
5.0
4.0
3.0
\D 11 .0
.. .j::o.
10.0
9.0
8.0
I 7.0
a.
6.0
5.0
4.0
3.0
Figure 57.
CSTR
'..
.' , I , ,1\ r' J" .
, , "--",''',,,,--".., I' 111'",4,,' ,
\,,,..'~I I,"~ 11..\ ,,1,,", ,,,,...I,,,,,,, ~, '. ,. II "
. , 1 '\V I" I I 1\ '''''-' .1' r"~ ,,-
, ! "J \JI,' ','I Mill " ~II" ,--I", 'f,I{""'" " "\.
" t", I I . I ~. ',', ,. 1 II,' , .I ,,' ..
" "', , ,', ',I, "I' 'I VI ,I
~ 'I 'S '," I " I, I, 'I ' I I 8 I I' . I,' I
I I; ,I I "I ~ I' ,,1111'11 I" I It III ,
, I : 011111: "": ,:I~ III: u ,: I
"' 11'1,1,11 , III, I I, '
" II \I,' I.' 1/1, I "
1 I II' I "'". "
I ,
CSTR A6
"
1\ .. ,-
\ I,' J "..... ~
I
I
,
,
EVENING
MORNING
- --
--~ ",)"''''''"',.",;.".,,'1 "\ ...
~ r " - ,. --" -\, I ,.. .r,
111 " ... "," V , .
III " I
1 II ,I I
I II ,I ,
, Q " ,
, , ,I I
I 1 ,I 1
I ,I I
I I ,
I
I
I
... ,'\ ., '\ 'I" "' (
, 1 ,I' ,11..1 I, I J. N 'I
, '1,1" 1'1 I, , I ' II
',,~IIII I, I' 'Ip' I,
I 'I j,' I I," 'I II I II
I 'I' ,I I I I I , I, "I 'I
I 'I ,I I ~ I I' I : 'I': 'I
I - ,I I I" I I I 1 'II, I
I I i I" ~ ,I 1 1 ~ 'I '
1 '" J I' " I
I' "
I' II
I
JUL
AUG
NOY
JAN
FEB
MAR
MAY
JUN
APR
DEC
86
EVENING
MORNING
- --
SEP
NOY
DEC
OCT
NOY
JAN
FEB
APR
JUN
AUG
OCT
JUL
MAR
MAY
DEC
pH Values in CSTRs A6 and B6
SEP
NOY
DEC
-------
\D
-------
\.0
0\'
:r:
a.
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
:r:
a.
11.0
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
Figure 59.
CSTR A8 EVENING
k_- I~,., MORNING
. -"\J--"
'..I"\J.Jo""\r ""--.,.,-,'
11\ ::., .,. - ~ I~ :
~:: :
, I
'~'
- --
--
i\r. ~J' 1 ", oIJ1~ '
I, 1'1 (,"/.1,... . #1 " ..,~
1,/ 11'1., 1" -, ..I(
, .I" I}r 'I I 1\ r " II' (, I ',. I .J' ~'"
, I , I " I' ,I 1 "~ \ (
,I , " 1 ',1 .' 1 1'", " I {I" "
,I ,~ I, ,~ 111 ~ ,1, I I I) I' ~", ~ I 1 ~ \ I
1 "', ' ~ 'II' " 'I~ I ' 1 "" ,I II t ~
, "If ~ I'UI ,I '. I I I ~ ,I ,I .,
I I 1 I' a' 1 II : 1 I I II P I
I I ,I t ' I II , , "
I I , I, II 1 1 1 I
I 1 1 \ 1 "
, I " II
\ ,I "
1
NQY
DEC
JAN
FEB
MAR
1 PR
MAY
JUL
AUG
SEP
OCT
NOV
DEC
JUN
cs~~: R 88 EVEJNI~G - - -
J ~vJJI-\ -- MORNING
- 1"''''"\, f-"'J\.. ~I\ , J\
" ,..I \ ~ IJ- I, I .1 , ~ '
1 I 1"\ ,--,,, I I 1 r ' 1# \ I 'J ~ h' r - r I
\, 1"-.\.1',; "I'i I, IAII~' ~l ': .J}~J'.~-'II"I/'!\"I\"\ /-/.,
L I' \ { I 1 I, "'~' I /1 I I'." " fo Iv II ,'i rl' "
I 1 , II I"",' I I ~ " I 111 I I I I I' I' L ( I
l\~ "11""",/,,,1 II I
\ I II I, i: II, I'll ~ I II II, I I' I '
. i' 'I ,/111'111 \',,1 1/ I,'
-v' I r'1 AI~: I,' ,III ,: II ,I I I /I~
Vv I' II , I \II
I , i" I' I! .
I IV .
Ah, JUN
~l
NOV
DEC
JAN
FEB
MAR
JUL
AUG
SEP
OCT
NOY
DEC
pH ValuE-:s in CSTRs A8 and B8
-------
1.50 * ~
I - 95% C.r.
'>;I * EARLY ~
~.
\0 ~ MID
c: C\1
t'1 S 1.25 QJ LATE
CI>
~ i
o 0
.
::I O;J:>o ~
0 0 c:
;j t'1 H\ b.O
(1' t'1 :E:
::r CI> c: :i. 1.00
en en 0
'O::r *
--0 en
......;j
DI 0. 0 ,--.......
(1' ~. ::r
CI> ;j...... M
....... \0 0
t'1 +
0 (1' 0
HlO'O
::r. ~
en N I-<: 0.75
I-<: ......
en S ...... '-"
(1'0 0
CI> :sIDl
\D ::I (1' .....
-....J ::r ~. b.O
0. en :s
ro 0
<:~f;j
ro CI> . r--i
...... DI 0
0 t'1 0 0.50
'0 ...... en
SI-<: .....
ro ......., DI ro
:s.. (1'
(1' I
~(1'
::r
::I t'1 r--i
0 ro
:s f1) ,.q
(1" ~
::ren 0.25
en DI U
::I *
--'0
S ......
,~.'~. i
o.:s
"""'\0
..
(1'
DI .....
.:S::l 0.00
o.ro
en
I.D
3 4 5 6 7 8
CSTR
-------
3.5
I - 95% C.r.
* EARLY * !
t'Ij 3.0 ili MID
1-" ili
\Q
r:: QJ LATE I
0; C\2
Ct> .8
0"1
I-'
. 0
\Ort"> 2.5
I-'.r:: ~
SSHI
0 Ct> ~ QD I
~ {/) r::
rt" 0 R
::ro::r
{/) 0 {/)
0;
~o;DJ
I-'roo. ,--.., 2.0
DJ {/) It) i
r? '0 ~ ~ *
Ct> 0 0
~ ::3 CI) +
0. 1-'-
0 1-" ~
HI ~ Ct> ~
\Q
{/) rt" ili
'< rt" 0; "--"
{/) 0 1-" 0 1.5
~ rt" '0 ~
00 Ct>N::r ttO !
S 0
S {/)
0.0'0 0
ro ~ ::r
< rt" !lI r--i
Ct>::rrt"
I-' {/) ro ili
0 1.0
'0 ~ 1-"
EI ro ~ ~
ro DJ
~ 0;. > E-;
rt"1-'tz1
'< 0
~o ~
... en
O"IDJ 0.5
rt"
S
0 rt"
~::r
rt"0;
::rCt>
{/) Ct>
~{/)
S DJ 0.0
1-'- S
0.'0
~I-' 3 4 5 6 7 8
.. 1-'-
~
DJ\Q
::3 CSTR
0.
-------
/ 0.6
I - 95% C.r.
'>:I * EARLY
.... ~ MID ~
t.O C\2
c:: 0.5
1"1 S Q] LATE
f1)
~
IV 0
.
\O(t';J>o ~
.... c:: QO
EI EI HI
0 f1) ~
=' en c:: S 0.4
rT 0 .6
='"o='"
en 0 en
1"1 *
~I"1OJ
I-' f1) en ~
OJ en ='" M
(t' '0 n
f1) 0 HI
~='1"1 +
a. f1)
0 .... f1) 0.3
HI=' ~
t.O a.
en 1"1 '-'"
~ rT ~
en 0 0
1.0 (t' ~ .-f
1.0 ro IV f1) QO
EI ....
Elt.O
a.o=," 0
ro =' (t'
<: (t' r--t 0.2
ro ='"....
I-'en=, !
0 ....
'O~r;; ~
EI f1)
f1) OJ 0
P 1"1 0
(t'1-'C/)r ~
~ *
~OJ ~
.. (t'
~(t' <4 0.1
='"
EI 1"1
0 ro i
P f1)
rT
=,"en
en OJ
EI
~'O
EI I-'
.... .... 0.0
a.P I I
~ t.O
.. 3
OJ 4 5 6 7 8
P
a. CST.R
-------
0.5
I - 95% C.r.
"'J * EARLY
~. .& MID
1.0 *
C C\2 [1] LATE
1"'\
II> S 0.4
0'1
IN
. U
~(I'»o ~
~. c
!::I a HI
0 II> ~ b1J
::J rn r.:
(I' 0 S
::ro::r
rn 0 rn
1"'\ 0.3
~I"'\(I'
1-'(1)0
I\) rn rr- ",--...... *
(I' '0 III
II> 0 I-' ~
~::J
0..0 +
0 ~. 1"'\
HI::J1O
<.Q III ~
rn ::J
'< (I' ~.
I-' rn 0 0 "--' *
a c1' a
II> NO 0.2
a !::I I)J .-t *
a 1"'\ b.D
0..00" ~
m ::J 0 0
<: (I' ::J
II> ::r r--I
I-' rn ~.
0 ::J
'O~
a II> »0
II> I)J tzj U
-:;$ 1"'\ 0
(1'1-'0 0
'
(I'll> m
::r ID
rn rn
III
~EI
EI'O
~. I-' 0.0
o..~.
~::J
... 1.0 3 4 5 6 7 8
III
::J
a. CST'R
-------
a.v
I - 95% C.r.
»j * EARLY
~.
o.a ~ MID
~
1"1 7.5 [!] LATE
t1)
0'1
~
.
51 0»' ~
0 0 ~ ,,-......
::s 1"1 HI ~
rt" 1"1 ~
::J" t1) ~
en en 0 '---" 7.0
'O::J" 0
~oen
.....::s .....
IlIc.t1' on
rt" ~. III
t1)'::s 0 0
..... o.a rt" *
t1) .....--i !
0 rt" 1"1
HI o~. !
III
en IV ..... 6.5
I-<: ro
en 51 ::s
'rt' 0 ~ .,..-!
t1) ::s a
I-' a 'rt" t1' H
0 ::J"t1>
I-' C. en 1"1 CJ.)
t1) en
<:~ ~
t1) t1) ~.
..... 1lI::S 0 *
0 1"1 6.0
'O""'g; ro
a I-<:
t1) ..... 0
::s .. '0 c:q
rt" C/)
. 0'1
III
a rt"
0
::s rt"
rt"::J"
::J" 1"1 5.5
en t1)
t1)
a en
~. III
C. a *
""''0
.. .....
~.
1lI::S
::so.a
c. 5.0
rt"
\O~.
a 3 4 5 6 7 8
t1)
en
CSTR
-------
First Order Rate Constant for DEP Degradation
10°
9
8 I
7 - C.I.
tzj
..... 6
lQ
~
1"1 5
(1)
0'\ 4
U1
.
...... t"Zj
() ..... 3
::r1"1
PJ en
:J rt'
:J
(1) 0
I-' 1"1 2
en c..
(1)
~ 1"1
~ ~ .....--....
c.. rr ~
(1)
IX! en I
PJ 0 H 10-
<: HI ~
(1) 9
f-' 1"1 c..
OPJro . "--" 8
NlQlQ
ro 1"1 .-4 7
c.. PJ ~
~ c.. 6
PJ
:(;rt
.......... 5
rr 0
::r:J
\D...... 4
U1;>o;
dPl-'
~
n 3
0 HI
:J 0
HI 1"1
.....
c..tJ
(1) t>:I 2
:J '"d
n
ro .....
:J
.....
:J ~
rT" t>:I
(1) n
1"1 0
<: en
PJ 10-
I-'
en
3 4 5 6 7 8
CS1'R
-------
3.0
+-'
:J
D-
C 2.0
"--
-f....J
:J
.B- 1.0
:J
o
0.0
I-'
o
!J.I
3,0
+-'
:J
D-
C 2.0
"--
+-'
:J
.B- 1.0
:J
o
0.0
CSTR A3
P04-P
Midday
Morning
-------
---------
- --
-- - -
~--~
--------------
"" .....
J.r- .....
Noy I Dee I Jan I Feb I Mar I Apr I May I ,Jun I Jul I Aug I Sep I Oct I Noy I Dee I
Time
P04,- P
Midday
Morning
CSTR. B3
-------
---------
~~ ~
/ ~~--
- --
---"---___0______---
Noy , Dee " Jan I Feb I Mar I Apr I MCJY' Jun I Jul I Aug I Sep I Oet I Noy I Dee I
Time
, Figure 66. Orthopho.sphab~ Output:Input Ratios in CS'rRs 1>.3 and 83
-------
+-'
:::J
Q...
c 2.0
"
+-'
:::J
.B- 1.0
:J
o
f-'
a
~
+-'
:J
Q...
c 2.0
"
+-'
:::J
.B- 1.0
:::J
o
3.0
0.0
3.0
0.0
CSTR A6
Midday
Morning
-------
P04- P
-------------
--------------
Noy
CSTR B6
Midday
Morning
--
-------
P04- P
------------
- - -, - - - - - - - - - - -
- --
Jul I Aug I Sep I Oct I Noy I Dee I
Figure 67. Or~hophosphate Output:Input Ratios in CSTRs A6 and 86
-------
3.0
-+-'
::)
0-
C 2.0
~
-+-'
::)
-8- 1.0
::)
o
0.0
......
0
V1 13.O~
c 2.0
~
-+-'
::)
-8- 1.0
::)
0
0.0
CSTR A3
Midday
Morning
-------
NOs- N
-------------
----------
May I Jun I Jul I Aug I Sep I Oet I Noy I Dee I
Time
CSTR B3
Midday
Morning
-------
NOs- N
------------------
- --
- --
- --
Figure 68. Nitrate~Nitrogen output:Input Ratios in CSTRs A3 and 83
-------
3.0 CSTR A6 N03- N
-+-'
:J
0..
C 2.0
~
-+-'
:J
(l 1.0 ------
-+-' ~
:J --""" -
0 '
.... - --
0.0 Noy
~
0 3.0
Q\ CSTR B6 NOs- N Midday -------
Morning
-+-'
:J
0..
C 2.0
~
-+-'
~
(l 1.0 - - - -----
-+-' -==-:::--
:::s
0
0.0 Noy
Midday
Morning
-------
- ~- -~ - -
--~-
Figure 69.. NitrateoorNitrogen output:lnput Ratios in CSTRs A6 and B6
-------
-t-I
:J
a..
c 2.0
~
-t-I
:J
.B- 1.0
:J
o
f-'
o
-....J
-t-I
:J
a..
c 2.0
~
-t-I
:J
.B- 1.0
:J
o
3.0
CSTR A3
Midday
Morning
NHa-N
-------
0.0
-------------
Nov I Dee I
3.0
CSTR B3
Midday
Morning
NHs-N
-------
---------------------------
0.0
Dee I
Nov
Figure 70. AmmoniaeNitrogen output:Input Ratios in CSTRs A3 and B3
-------
3.0
CSTR A6
NHs-N
Midday
Morning
-------
+'
:J
Q..
C 2.0
""-
+'
:J
E- 1.0
::s
o
----------------
--------------
/'
1\
\;~_Jj \~---_J-_J~~-------------'
0.0 I Nay I Dee I Jon I Feb I Melr ! AprF'May T Jun I Jul I Aug I Sep I Oct I Nov I
Ti rn e
Dee I
f-'
a
co
3.0
CSTI~ B6
NI:I3 - N
Midday
Morning
-------
+'
:J
Q..
C 2.0-
i 1.J - - - - - - - - - - - - - - - -- - - - - - - - - - - - - -
<5 J 'v-_/",---~~,. ' i'.
I .- . ~. ~ ~ '''--------''''- -K- ~ ~ - - ........-.-J./ - --.."---:---
O.O-"-N(~;-;-6~~c ~=J~i-'-;e~~~~~~;T~;p~~~y I l~:n ~-~u~-I-~:g~l~e; r~~~'1 N:V -I Dee-'
Tirne
Figure 11. Ammonia-Nitrogen Output:Input Ratios in CSTRs A6 and B6
-------
5.0
C\1
8
C)
~4.0
Q[)
q
...-.....
cd
I
r--i
~
C)
'-;;; 3.0
.-I
Q[)
o
r--i
2.0
0.0
Figure 72.
C) A
~ B
6
(!)
~
(!)
1.0
loglo(TDP)
C)
(!)
~
A
A+B
B
Linear relationship between aufwuchs chlorophyll a
concentration and total dissolved phosphorus input
concentration for Channel A, Channel B, and Channels
combined.
109
2.0
/Lg!l
A and B
-------
5.0
(9 A
~ B
C)
C\2 ~
S C) ~
C)
0 ~
~4.0
bD
~
~ ~
cd C)
I
r--I
~
CJ
~3.0
..-.1
OD
0
r--I ~
C)
2.0
0.0
1.0
IOg10(11DP)
2.0
fLg/l
Figure 73. Curvilinear relationship between aufwuchs chlorophyll a
concentration and total dissolved phosphorus input
concentration for Channels A and B combined
110
-------
CL
-f-I 250
:J
0...
-f-I 200
:J
.0
~ 150
Z 100
-f-I
:J 50
0...
-f-I
:J
o
f-'
f-'
f-'
CL
-f-I 250
:J
0...
-f-I 200
:J
o
~ 150
z 100
-f-I
~ 50
-f-I
~ 0
CSTR A3
MIDDAY
MORNING
--------
CSTR B3
MIDDAY
MORNING
--------
-~-,~--- J~~
-----~-------'~------~~~ ~--- ~---- ,--
NaY' DEC I JAN I FEB I MARl APR I MAyTJUN I JUL IAUG I SEP I OCT I NOyl DEC I
Figure 74. Dissolved Nitrogen:Phosphorus Output Ratios in CSTRs A3 and 83
-------
[L
-f-J 250
::J
0...
-f-J 200
::J
o
~ 150
:z 1 00
-f-J
::J 50
0...
-f-J
::J
o
.....
.....
N
[L
-f-J 250
::J
0...
+-' 200
::J
o
~ 150
:z 1 00
-f-J
::J 50
0...
-f-J
::J
o
CSTR A6
MIDDAY
MORNING
-------.-
o
csrrR B6
MIDDAY
MORNING
--------
-
""" J --"'\ .r--
J./ V \ -'= J - """-
v .....
./'-
o
Figure 75. Dissolved Nitrogen:Phosphorus Output Ratios in CSTRs A6 and B6
-------
Table 1.
Elemental concentrations in AEcoS.
. .. ..-
. - .. . . . - - . - - - - - -
. . - ~ . - . . -
- . -. . - .
Macronutrients
mg/1
Micronutrients ug/1
.. . . .
B 3100
Mn 11.1
Zn 14.9
Fe 31.4
Co 0.329
CU 0.00354
Mo 2.75
EDTA 440
.. . . . . - - .
. - .. - - . . - - . - - . - - -
p
0.15
Si
2.94
HC03
34 .50
Mg
1.33
K
2.77
N
2050
Ca
4.82
S04
8.66
Na
18.03a
26.33b
C1
17.53 c
30.40d
43.27e
. . . . - . . . . . - . . . . . . -. .
- - - - -.-' . .. - -. . - . . .
. . - . . . . . - - . . - - . . . -
aAmount added from Na salts
bTota1 Na after addition of NaOH for pH adjustment in CSTR 2
cAmount added prior to acid/base addition (inorganic chlorides plus HC1)
dAmount added through the addition of 0.610 meq/l HC1
eAmount added through the addition of 0.968 meq/1 HC1
113
-------
Table 2.
Nutrient quantities in 44 1 of macro-nutrient concentrate (drip
rate of 4.0 m1/min into CSTR 3).
- - . . . . . - - -
. - - - - - . - - .
- . . - - . .
- - - - . - . . -
Concen ti: a te . !
ATnount
Na2Si03 . 9 H20
113.56 9
*Na2HP04 . 7 H20
0.494 9
NaHC03
181.4 9
Concentrate. II
Amount
MgS04 . 7 H20
51.52 9
K2S04
23.58 9
*NH4N03
2.73 9
CaC12 . 2 H20
67.48 9
Micronutrients
362.8 m1 stock
- . - - - .
. - - - - .
- - - - - -
. . - - - . - -
- - - - - . . . . . . . -
*10% of quantity was dripped into CSTR 3, 90% into CSTR 6.
114
-------
Table 3.
Nutrient quantities in 1.0 1 of micronutrient concentrate.
- - - - - . - - - - - - - . - - -
. - . .. - - . . . . - - - - -. - - - - - - . . - . . . - . . .
- . . . - . -
H3B03 1.855 9
l>1nC 12 . 4 H20 0.420 9
znCL2 0.327 9
Na2EDTA . 2 H20 6.0 9
COC12 . 6 H20 0.014 9
CUC12 . 2 H20 0.001 9
Na2Mo04 . 2 H20 0.073 9
FeCL3 . 6 820 1.599 9
HCl (12 M)a 220 ml
- . - - . . - . . - . . - - - - - - - - . - - ~ - - - - - - - - - - - - - - - - - - - - - . . - - - . - . . - - - - - - - - ~ - - - - . . - - - - - - - -
aAmount of RC1 required to neutralize Na salts of Concentrate I after
addition of Concentrates I and II to channel flow
l1S
-------
Table 4.
Major sampling dates for DEP experiment.
. - - . ~ - . - . - - - .
. - - . . . . - - - - . . . - - - - - -. - . - . - - - . - . . -. - - - . . .-
- - . - - - . .
. . - - . - - .
Date
Number Samples
eSTRs samp1ed1 per eSTR
. . - - . - . - . - - - - - - - . . . . . - - . - - . . ~ - - . . . . - - - - - . - - . - - - - - - . . . -
Parameter
Measured2
- - - . - - - .
. - - - .. -
11/20/79
1/8/80
1/22/ 80
2/5/80
2/20/80
5/27/80
8/5/80
8/12/80
8/19/80
8/26/80
9/9/80
9/16/80
9/23/80
9/30/80
10/7/80
10/14/80
A3-8, 83-8
A3-8, 83-8
A3-8, 83-8
83, 86
83, 86
A3-8, 83-8
A3, 83
A4, 84
A5,- 85
A6, 86
A7, 87
A8, 88
A3, 83
A4, 84
AS, 85
A6, 86
1
1
1
4
1
1
3
3
3
3
3
3
3
-3
3
3
I
f
2-4
1-5
1-53
1-5
1-5
1-4
1-6
1-6
1-6
1-6
1-6
1-6
1-6
1-6
1-6
1-6
- . - - . .
- - - - - - . . . - - - - - - . -. . . .. - -. -
. -. -.- - . - - - . -
. . . . - - - -. - - - - . - - - - - - . . . --
1200 ug/1 DEP introduced into each pair of eSTRs for seven days prior to
sampling dates between 8/5/80 and 10/14/80
2parameter code: 1 - aufwuchs TOe; 2 - suspended NH3-N, N03-N,
N02-N, DKN, P04-P, TDP, DOe, Toe; 3 - DO, pH; 4 - aufwuchs AFDW,
ATP, bacteria plate counts; 5 - aufwuchs algal enumeration and
identification; 6 - toxicant analysis
ehl ~,
3eSTRs 3 and 7 samples not taken for algal identification and enumeration
116
-------
Table 5.
Steady-state concentrations of DEP (ug/l) due to
sorption/volatilization, photolysis, and hydrolysis of a mean
input concentration of 191 ug/l DEP (s = 6.0, n = 20).
. - . - - -. . . - " - - - - - . . -
- - - - - --
Sorption/Volatilization
Photolysis
Hydrolysis
. . . - . - . . -
- - - - . . -. . - . - - -
. . - . . . - . - -' -
Channel
CSTR
-
x
s
n
-
x
s
n
-
x
s
n
- -. - - - . . - - - - - - . - -. - - - - - - . . . - - . . . . - . . - - - . - - - - - - . - - - - - . - - - - - - -
. - - . - --
A 1 192 5.3 10 187 3.4 8
A 2 192 4.4 10 178 4.4 8
A 3 188 5.3 10
A 4 188 4.3 10
B 1 193 6.2 10 190 4.3 8
B 2 190 6.7 10 182 3.9 8
B 3 185 5.4 10
B 4 189 5.6 10
- - . - - . - - - - . - -' - - - - - - - . - - - - - - - - . . - . . _.. . - . - - - - . - - - - - --
- - - - - - - - - . - --
- - - . - - - - - - --
Note:
Study conducted in the absence of biological growth
117
-------
Table 6.
concentrations (mg/l), % degradation, and firsteorder degradation
rate coefficients (kl) of DEP in AEcoS.
. . . - - .
- . - - ~ . . - - - - . - - .
- . . . . . -
- . - - - - . - - -. -
. - - - - . - - - . . - - -
-. - - - - - - .
concentrationl
% Degradedl
kl(hr-l)1,2
- - - - - - -
- - - . -. . - - . -
Channel
CSTR
-
x
s
-
x
s
x
s
- - - - - . -
. . - - - - - - .
- - -. - . . . . -
- - . -. . - . - . -.
. - - - - . - - . . - -
. - - - - - -
- - - . - - . -
- . . - - -
A 3 0.043 0.006 77 3 0.294 0.051
A 4 0.049 0.006 73 3 0.251 0.043
A 5 0.116 0.006 37 3 0.057 0.007
A 6 0.073 0.005 60 3 0.148 0.016
A 7 0.054 0.004 71 2 0.232 0.023
A 8 0.066 0.008 64 4 0.173 0.029
B 3 0.046 0.007 75 4 0.272 0 .051
B 4 0.050 0.004 73 2 0.245 0.024
B 5 0.118 0.003 36 2 0.054 0.003
B 6 0.061 0.004 67 2 0.194 0.018
B 7 0.038 0.002 79 1 0.361 0.029
B 8 0.019 0.002 90 1 0.777 0.077
- . - . . - -. . -. - - . - - ~ . -
In = 7
2kl calculation based on a DEP mean input of 0.194 mg/l (s = 0.006, n = 22)
118
-------
Table 7.
Estimated chlorophyll ~, ATP, and bacteria for CSTRs 4 and 8 in
sediments, suspended matter, and aufwuchs during DEP additio~s.
Numbers in parentheses indicate percentage of total found in each
phase.
.. - - . - - - -
- . - - - - .
- - - . - . - - -
Sediment1
Suspended
l>.ufwuchs2
. . - . . . -. - - - . . . -
Chlorophyll ~ (ug/l)
CSTR 4
CSTR 8
34.8 (8.9)
120 (18.6)
6.6 (1.7)
41.0 (6.3)
351 (89.4)
484.8 (75.1)
ATP (ng/l)
CS'!'R 4
CSTR 8
2.8 x 103 (24.5)
1.6 x 104 (18.7)
1.4 x 102 (1.2)
1.2 x 103 (1.4)
8.4 x 103 (74.3)
6.9 x 104 (79.8)
Bacteria (cells/I)
CSTR 4
1.8 x 107 (12.3)
2.8 x 107 (17.4)
3.4 x 108 (81.0)
. - - - - - -
. . - - - - - . - . .
- . . . - - .
. . . - . . - . - . - . - .
lconverted from a dry weight
weight of 3000 grams for the
dividing by CSTR volume
(mg) to a volume (1) basis by estimating a
upper 25% of the total sediment mass and
2Converted from area (cm2) to volume (1) basis by estimating a total
CSTR surface area of 3.0 m2 and dividing by CSTR volume
119
-------
Table 8.
Calculated SIMI values for aufwuchs algal assemblages during DEP
additions.
- ~ ~ . - - - - - - -
~ - . ~ . - . - - - - - . ~ - . - . . . . -
. . - . . - -. -.. . . - - -. . - -. . . - . -. . - --
Channel A Channel B
- . . - .. - . -. - -. - - - - - . - - - - . . . - . - - - - - - - -. . - . - - - . - - . .- - .
CSTR Comparison
All Taxa Bluegreens Greens All Taxa Bluegreens Greens
- - . . -. . - - - - - - - - . - - - -... . - . . . - . . . - - - . . " - - . - . - -. . -. . - - - - -.. . .. - - . - - .
3..4 .36 .29 .92 .85 .99 .41
3..5 .23 .27 .85 .86 .99 .23
3..6 .31 .28 .67 .67 .71 .17
3-7 .57 .80 .62 .32 .41 .17
3..8 .50 .79 .59 .09 .09 .19
4-5 .69 .99 .96 .94 .99 .81
4..6 .81 .99 .59 .59 .71 .24
4-7 .51 .48 .55 .34 .43 .23
4~8 .40 .27 .52 .09 .11 .49
5-6 .57 .99 .75 .62 .70 .32
5..7 .60 .46 .72 .31 .36 .25
5-8 .62 .24 .70 .02 .02 .47
6-7 .76 .47 .99 .39 .37 .95
6..8 .69 .25 .99 .13 .13 .72
7..8 .98 .96 .99 .69 .93 .78
- . .. ... . . . . - - - - - - - - - - - - -. - - . . . -. - . . - . - . .
120
-------
Table 9.
Groups of CSTRs having similar aufwuchs algal assemblages during
DEP additions - Channel AI.
All Taxa
CSTR
Grouping
3
8
7
6
5
4
-. - - - . -" - . .
- . - . - . - - . . . - - - .
. - -. . . . - .
- . . -. - . . . - -
. . . . . - - . - - . -. . - . . - . - -' - . - .
Bluegreens
CSTR
Grouping
4
5
6
7
8
3
. - . - - . .
. .. - - . . .
- . - - -. .
- - . - - -. . .
Greens
CSTR
Grouping
3
4
5
6
7
8
- . - . - - . -. . . - -. . . - -. . - - - - - - . - . - - -
- - - . - - . - - . - - - .
- . - - - - - - - . -. . - - - - - - - . . - - - - . . - - - - . - - - - - .
. - - - - . - - - - - - . .
IHorizontal bars group comparisons with SIMI values> 0.50
121
-------
Table 10.
Groups of CSTRs having si~ilar aufwuchs algal assemblages during
DEP additions - Channel B .
. . - . -.- - - - . . . . . - . - . - - - . - -. . . . -. . - - . - . - - - . . - .
. . . . ~ . - - - . - . . . . - .
. - . . .. ..
All Taxa
CSTR
Grouping
3
4
5
6
7
8
. - '.- -. - - . . - - - - . - - - - . - . - . -. . - - ..
Blue Greens
CSTR
Grouping
3
4
5
6
7
8
. - - - - - - - .. . - . . - - - - . - - - - . - - .. . . .
Greens
CSTR
Grouping
3
4
5
6
7
8
- - --. . - . . . . -
. . - . - . -. . -
.. - - - .
- - . - - - . - - . - - - - - . - . - - - - - - - . . - -
- -. . . . --
lHorizontal bars group comparisons with 81M1 values> 0.50
122
-------
Table 11. Relative abundance (%) of aufwuchs algal species compr ising > 1.0% of the total density on
1/22/80.
CSTR
Aufwuchs Algal Species 3A 3B'. 4A 4B 5A 5B 6A 6B 7A 7B 8A 8B
BLUE..GREENS
Chroococcus dispersus 23.5 1.7
Oscillatoria sp. 9.0 26.5 3.9
Lyngbya subtilus 1.8
Phormidium sp. 90.7 81.0 47.6 75.2 88.0 23.6 8.1 55.4 16.5
Phormidium subfuscum 1.2
Aphanothece nidulans 2.3
Aphanothece saxicola 3.4 2.8
GREENS
Ankistrodesmus falcatus 2.8
...... Scenedesmus denticulatus 5.0 8.0
N
(,N Scenedesmus dimorphus 1.5 20.3 56.1 32.2 46.8
Scenedesmus obliquus 3.4 3.1 1.2 4.4
Scenedesmus bijuga 2.7
Stigeoclonium sp. 1.2 1.5 2.9
Oedogonium sp. 10.4 2.6 1.2
Coelastrum microporum 1.0 3.8 1l.2 6.9 33.3 16.3
DIATOMS
Nitzschia palea 1.6 2.9 3.7
Synedra ulna 1.0 2.3
Synedra incisa 2.1
Synedra rumpens 1.4
Achnanthes minutissima 1.3 3.8 3.4
% of Total Density 95.2 96.2 98.3 95.2 95.7 98.7 95.1 95.0 97.7
CSTRs 3A, 7A, and 7B not sampled
-------
Table 12. Relat:i ve abundance (%) of aufwuchs algal species campr ising > 1.0% of the total density
during DEI' experimentation (8/5 0;- 9/16/80).
CSTR
Aufwuchs Algal Species 3A .313 4A 4B SA 5n 6A 6n 7A 7B 8A 8n
BLUEeGREENS
Chroococcus dispersus 62.4 3.8 3.2 6.4 29.2 26.3 27.1 81.6
Chr oococe us minor 1.6 3.0 1.2 5.1 4.8 2.7
Oscillatoria Spa 2.7 36.8
Oscillator ia subbrevis 1.8 3.0
Lyngbya subti1us 2.8
Phormidium Spa 12.7 55.9 34.1 35.7 3.7 38.6 45.0 36.8 15.5 10.1 7.0 1.6
GREENS
Chlamydomonas g10005a 8.1
Dictyosphaeriurn ehrenber.gianum 1.3
Ank istrodesmus falci.it:us 1.2 2.5 3.8 3.4 3.6 4.4 10.7 24.3 1.1
~ Scenedesmus denticu1a\:us 1.5
N Scenedp.smus dimorphus 1.5 1.3 3.3
"""
Scen~"?desmus bijugil 6.7 15.4 35.6 2.5 43.3 46.5 8.4 49.4
'l'etr aedr 011 min imum 1.7 3.9 2.4 12.1 1.1
Oedogonium s1'. 1.2 12.7 21.3 24.4 17.6
Cosmar iurn meneghinii 2..0
Cosmarium subreniforme 1.5
staurastrum Spa 4.5
Coe1astrum microporum 9.4 23..3 10.8 27.1 2.5 5.1 2.5 6.3 1.1
Zygnema Spa 13.8 10.0 2.7 5.8
DIATOMS
Ni t zsch ia palea 5.6
Navicula eonfervacea 5.7
Navicula minima 2.5
Gomphonema parvulLUn 2.0 4.4 2.6 2..2 1.4 2.0
Cyrnbella minuta 1.5
Achnanthes minutissima 3.9 2.2
% of Total Density 94.4 95.2 93.6 97.7 98.2 91.4 96.1 94.9 97.9 95.0 94.1 96.3
-------
Table 13.
Calculated HI diversity and J evenness values for aufwuchs algal
assemblages during DEP additions.
.. -. -'
Channel A
Channel B
. ~ . - . . . . - . . . . - -
. . . . ~ . -
. - . - . -. - -. . . - . . " - --
CSTR
HI
J
HI
J
- - - . . . . - .. - . - - - - - - . - - - - -. - . . - . . - . - - - - - - - . - - - - - - - " . -. -
- - . -. - - - . -. - . - '.0' - - ... -
3
1.97 0.47 2.72 0.70
2.69 0.63 2.88 0.74
2.27 0.58 3.14 0.68
1.69 0.47 2.68 0.63
1.97 0.57 3.13 0.74
2.16 0.53 1.27 0.32
4
5
6
7
8
- . - - - - - - . . -
- - - . . - . . - - . - -.. - - - - - . - - - - - - - - - - - - -. -- - - - - - - . - - - - - - - - - - - -. - - . - - - . - .
125
-------
Table 14.
Results of Newman-Keu1s Range Test for aufwuchs biomass during DEP
additions (a = 0.05). Means expressed as lo910(x + 1), n = 6.
- -. . . - .
. . . . - - . -
- - . - - - - - - - - - . . . -
Chlorophyll ~
CSTR
Mean
Grouping
6
1.29
. ~ ~ . - - - - - - ~ - ~ - . . * .
7
1.26
ATP
CSTR
Mean
Grouping
6
2.98
AFDW
CSTR
Mean
Grouping
6
0.38
7
2.77
8
1.13
8
2.74
7
0.37
~ . - - . . ~ - -
8
0.28
- . - . - -
. - . . . . . - - - - - . - - . ~ ~ . - - . - .
3
0.76
3
2.21
3
0.21
4
0.51
5
0.12
4
1.84
5
1.27
4
0.17
5
0.06
. - - - - -
-. - . --
7
0.13
- - - . - - - - - - . . . ," - - .-
. - - - - . . - - --
3
0.05
7
6.76
. - - - - - - --
- . . . . . ~ - . .
- .
. - . - . - - - . - . - - -
TOC
8
0.12
4
0.04
5
0.02
CSTR
Mean
Grouping
6
0.17
Bacteria
CSTR
Mean
Grouping
6
7.21
3
6.51
8
6.41
4
5.84
5
5.81
- - . - - - - -
126
-------
Table 15.
SIMI values comparing aufwuchs algal assemblages on 1/22/80 to
DEP data.
. + -. - - - - -
. . . - . - - - . -. - - - - - -
- - - - . .-
Channel A
Channel B
- - . - - - . . ~ . - - - . - + - . . - - . . . . -
- . - . - - . - . - . . . .- ~ . . - - . .
CSTR
Dates
Compared
All Taxa
B1uegreens
Greens
All Taxa
B1uegreens
Greens
- - . - . - - . - - ~ - ~ . . ~ - -. - . . - . . - . - - - - - . . .
. . - . - - . .. . . - - - - - . - . . . .
. . - " - - . .. . .- - - - - . - . -
3a 2/5~8/5/80 .95 .99 .04
4 1/22-8/12/80 .77 .99 .78 .83 .89 .82
5 1/22-8/19/80 .12 .99 .51 .87 .99 .48
6 1/22-8/26/80 .43 .95 .10 .42 .90 .12
7a
8 1/22-9/16/80 .12 .24 .03 .01 .02 .25
. - - - - - . . - . - - . - - - . . . . . . - - - - . . . - - - - - - '.' . . . - . - -. - - . - . - -' - -. - - - - - . . - - - - - . - . - . . - -
aCSTRs 3 and 7 not sampled 00 1/22/80
-------
Table 16.
Calculated product moment correlation coefficients for aufwuchs
structural parameters during DE? additions.
. . . - . . -
- - - - . . . - . - -. . - . . . . . . - . - - . . - - . .
- - - - - . - - . . . .
Chlorophyll a
AP Dvi
ATP
TOC
. - - - - - . . . . . .. .-
. . . . - . . . . A' . - . . . .
. . . - . - -
- - .. .
Chlorophyll ~
1.00
0.91
0.85
0.88
AFDW
1.00
0.87
0.84
ATP
1.00
0.70
TOC
1.00
. . - . . - . ~ . . . . . - . . . . - - .-
. . - . - .. . . - .. . . - - - -
- - .. ..
n = 60, all correlations significant (p < .001)
128
-------
Table 17.
concentrations of trace meta1s* in aufwuchs from AEcoS the week
of December 2, 1980.
. - - - - - - - - ~ . - . - - - - - . - .
- . - - - - . - - . . . .
. - - - - . . - - - - - - - . - - ~
. - - - . - -
- - . . - . ..
ug/ 9 dry wt
mg/g dry wt
- - . . - " - . - - - . -. . -. - - .-
- - . . - . .. - . . - . . - . .. ~ . . ~ - - -
CSTR
% Ash
Cu
Zn
Mn
Ca
Mg
Fe
Na
. - -.. - .. . . .. - - - - -
. . - - -.. - - . - - - .. . - - - - - - - - - - . - . - .-
- - - . . ... .. . - -. . - - . - - - - - - . " - - - -
A3 7.99 11.5 6.3 519.3 5.6 0.8 0.3 1.3
A4 10.12 9.9 7.3 238.1 4.5 0.5 0.4 1.3
A5 9.91 8.2 6.6 220.4 5.4 0.4 0.5 2.1
A6 9.00 27.9 58.1 66.8 5.6 4.8 0.8 1.5
A7 7.59 31.0 44.7 103.4 10.9 4.0 0.6 1.8
A8 10.35 37.3 85.5 185.3 9.5 3.4 1.4 1.7
B3 7.89 14.4 18.1 293.8 8.0 1.3 0.4 2.8
B4 14.78 13.4 8.6 172.4 6.6 0.3 0.3 2.5
B5 7.92 10.3 18.0 945.3 6.9 1.0 1.3 3.3
B6 6.13 26.7 29.8 60.2. 5.9 5.0 0.4 2.4
B7 6.20 36.7 43.6 64.4 8.3 3.4 0.7 1.7
B8 22.81 23.7 44.9 378 .5 7.3 3.3 1.6 1.6
. - - - - - . - - - - -. . . . - - - - . . - - - -. - -.. - - . - . . - - - . - - - - - - . . . - - -. - - . - . . -. ' .. . - - - - . -. . - -. .
*Co and Mo were below detection limits
129
-------
Table 18.
Mean chemical concentrations (mg/1) in CSTRs over major sampling
dates (see Table 4).
. . - - . . . . - . ~ - . . ~ . . . . . - . - ~ - - ~ - - ~ - - - - . - . - - . - . ,- - . - - . - - . - ~ . - - - - . - . . - - - . - - . .
DOC - . - . ~~4 -p - - ~O~~N - - - . NH3-N
- - - - . - . . - . . - - . - . .
CSTR
-
x s x s x s x s
. - . - - . - -. - - . - - - . - . - - - - - - . . - - - - - - . - - - - - - - - -. - - - - - .. - - - - - - - - '. - -. - - - - - - - - - . - -
3 1.8 0.2 0.004 0.000 0.06 0.02 0.035 0.003
4 1.2 0.2 0.003 0.000 0.02 0.01 0.012 0.004
5 4.0 0.0 0.004 0.000 0.01 0.00 0.014 0.001
6 2.1 0.0 0.029 0.027 1.06 0.16 0.318 0.087
7 2.7 0.8 0.044 0.037 1.06 0.07 0.253 0.130
8 2.8 0.7 0.027 0.014 0.82 0.02 0.212 0.046
- . . - - - - .
- . - - - - .
- . - . - . - . - - --
- - - . . - - - - - - - -. - - - - - - - - - - - - . . - - - - .
. - . . - - . - . -.
130
-------
Table 19.
Total dissolved phosphorus and aufwuchs chlorophyll ~
concentrations foe each CSTR in AEcoS.
- - . . - . . . . ~ . . . - - . - - - - . . ~ - ~ ~ . . - - . - - - - - - - - - - - . - - - . - . . ~ - . -
TDP (mg/1) Ch1_i- ~~g/cm2)
. - - - - - . - - - -. - . .
peeiod Colonized CSTR
-
x s x s
. - - - - - - - - - - - - . - - - - - - - - - - - . - - . . . - - - - - - - - - . - - . - - - . . - - - - - . - - - - - - . - . - - . - - . - . . - - . -
7 /7/80~8/5/80 A3 0.017 0.002 4.2 0.7
7/7/80-8/5/80 B3 0.017 0.002 5.7 2.9
7 /14/80~8/12/ 80 A4 0.005 0.001 1.4 0.5
7/14/80-8/12/80 B4 0.005 0.001 3.4 0.4
7 /21/80~8/19/ 80 A5 0.004 0.001 0.3 0.1
7/21/80~8/19/80 B5 0.004 0.001 0.4 0.1
7 /28/80~8/26~80 A6 0.134 0.006 16.8 5.3
7/28/80~8/26/80 B6 0.135 0.006 21.6 8.4
8/11/80..9/9/80 A7 0.021 0 . 0 08 17.1 5.3
8/11/80-9/9/80 B7 0.042 0.012 18.5 6.3
8/18/80~9/16/ 80 A8 0 .024 0.005 14.2 0.8
8/18/80-9/16/80 B8 0.069 0.011 10.9 1.2
8/25/80..9/23/80 A3 0.017 0.002 6.4 0.3
8/25/80~9/23/80 B3 0.017 0.002 5.2 1.3
9/1/80~9/30/ 80 A4 0.004 0.001 2.9 0.4
9/1/80~9/30/80 B4 0.004 0.001 1.6 0.4
9/8/80~10/7 /80 A5 0.003 0.001 0.8 0.1
9/8/80~10/7/80 B5 0.003 0.000 0.6 0.0
9/15/80~10/14/80 A6 0.133 0.006 37.5 18.0
9/15/80~10/14/80 B6 0.133 0.006 14.8 7.8
. - - - -. - - - - - - - - .. - -.. - - - . - - - - - - - - - - - -. - - -. -... - - - - - - - -.. - - - -. - . - - - - - - - - - - -. - --
131
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Table 20.
Analyzed mean N:P input and output ratios (by weight) for
11/16/79 c 12/1/80.
. . ~ . . . . . .
. - . . . . . -. . . - .. . .. . - . . .. . - . .. - - - - - -
. . . . -. - -. . . . - . . - . --
- - - - - - -
CSTR
Sample
Time
N:P Inp~t Ratios
x
N:P Output Ratios
- - - . - . - . . - -. .. - . - . . . - - - - -
Lowes t
Highest
-
x
s
n
- -. . . - - - - - - - - - . .. .
. . - - - . . . - - . - - - .. - -. -. - . -. -. . .
A3
83
morning
morning
14.82
14.76
14.82
14.76
16.93
16.77
16.93
16.77
. - . . . - - - - - - . . . . . - -
A3
83
midcday
midcday
-' - - . - . . .
. . . . .. -. . . - . .
4.00
3.60
59.00
75.50
22.34
24.62
12.69
12.83
78.64
45.07
59.12
40.15
.. -.. - - - . .
12.99
14.53
9.05
6.59
57.34
33 .18
37.58
15 . 21
- - - - - . .
52
52
51
51
52
52
51
51
A6
86
morning
morning
A6
86
midcday
midcday
1.20
2.00
43.20
30.00
18.03
18.56
262.80
191.00
13 . 83
20.66
152.90
75.60
. - . . . - - - -
- - . - - . .
132
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Table 21.
Replicability and aiel pattern student's t~test resultsl.
A versus B Morning versus Mid..day
Nutrient Timeblock2
CSTR 3 CSTR 6 CSTR 3 CSTR 6
NH3..N a n.s. n.s. * *
b n.s. n.s. * n.s.
c n.s. n.s. n.s. *
d n.s. n.s. * *
N03..N a n.s. n.s. n.s. 005.
b n.s. n.s. * n.s.
c n.s. n.s. n.s. n.s.
d n.s. *' * n.s.
P04"P a n.s. * * *
b n.s. n.s. * n.s.
c "* n.s. n.s. n.s.
d n.s. * n.s. *
In.s. = not significant at
Cf.= 0.05;
Ok. . c. .L.
slgnl.1-l.can... at Cf. = 0.05
2a = 11/27/80 to 2/20/80
b = 2/21/80 to 5/20/80
c = 5/21/80 to 7/20/80
d = 7/21/80 to 12/5/81
133
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Table 22.
Results of Student's t-test comparing aufwuchs biomass parameters
in Channels A versus B during DEP additions.
- - - . . .. . . - - . -
. - . . -. . . .
- . - - .. - - .. . . . . - . -
. - - . . . - . - - . . . -
- - - - -... . . - -. - - - -
CSTR
- - . - . - - . . - - - - . .
. . - - - - .. . - - - - . . . - - - -
. - . -. - .
Parameter 3 4 5 6 7 8
Chlorophyll a n.s. * n.s. o.s. n.5. *
ATP n.5. * * n.s. n.s. *
AFDW n.s. n.5. n.5. n.s. n.s. n.s.
Toe n.5. n.5. n.5. n.5. * *
Bacteria * * * * n.5. *
- - ,' - - - -
* = significantly different at 0.= 0.05; n.5. = not significantly different
(n = 6)
134
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Table 23.
SIMI values comparing aufwuchs algal species composition in
Channels A and B during DEP additions.
SIMI Value
CSTR All Taxa Bluegreens Greens
3 .31 .33 .09
4 .86 .99 .66
5 .33 .99 .54
6 .54 .74 .11
7 .60 .97 .34
8 .48 .96 .15
- - . . - . - . . . . . . .. - - - - - - - - . - . . - . -. .. . . . . .. -. - - .
135
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