&EPA
United States
Environmental Protection
Agency
Environmental Research
Laboratory
Corvallis OR 97330
EPA-600/9-79-021
July 1979
Research and Development
Bibliography of
Literature
Pertaining to the
Genus
Selenastrum
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7. Interagency Energy-Environment Research and Development
8. "Special" Reports
9. Miscellaneous Reports
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/9-79-021
July 1979
BIBLIOGRAPHY OF LITERATURE PERTAINING TO
THE GENUS SELENASTRUM
by
Amy A. Leischman, Joseph C. Greene, and William E. Miller
Freshwater Systems Division
Corvallis Environmental Research Laboratory
Corvallis, Oregon 97330
CORVALLIS ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CORVALLIS, OREGON 97330
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DISCLAIMER
This report has been reviewed by the Corvallis Environmental Research
Laboratory, U.S. Environmental Protection Agency, and approved for publica-
tion. Mention of trade names or commercial products does not constitute
endorsement or recommendation for use.
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FOREWORD
Effective regulatory and enforcement actions by the Environmental Protec-
tion Agency would be virtually impossible without sound scientific data on
pollutants and their impact on environmental stability and human health.
Responsibility for building this data base has been assigned to EPA's Office
of Research and Development and its 15 major field installations, one of which
is the Corvallis Environmental Research Laboratory (CERL).
The primary mission of the Corvallis Laboratory is research on the ef-
fects of environmental pollutants on terrestrial, freshwater, and marine eco-
systems; the behavior, effects and control of pollutants in lake and stream
systems; and the development of predictive models on the movement of pollu-
tants in the biosphere.
This report is a supplement to the 1978 Selenastrum capricornutum Printz
Algal Assay Bottle Test. It reflects the varied and world-wide application of
algal assays in water quality assessment and management.
J. C. McCarty
Acting Director, CERL
m
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PREFACE
The 1976 Experimental Use of Algal Cultures in Limnology Symposium (held
in Norway) centered around the use of algae as sensitive indicators to define
the stimulatory and/or inhibitory effects of pollutants entering aquatic
ecosystems.
The necessity of a standardized test protocol to insure comparative
results throughout differing geographic regions and in waters of varying
quality was recognized. The Algal Assay Procedure Bottle Test (U.S. EPA,
1971) was cited by Chiaudani and Vighi (1978) as the only algal growth test
that had been evaluated and refined enough to be a reliable and reproducible
test. This test protocol was endorsed into the 14th edition of Standard
Methods for the Examination of Water and Wastewater (APHA, 1975). A revised
and updated document, The Selenastrum capricornutum Algal Assay Bottle Test
was published by Miller, Greene and Shiroyama in 1978. This document is the
foundation of the algal assay growth potential and effluent testing protocols
currently in the ballot process for inclusion in the American Society for
Testing and Materials Annual Book of ASTM Standards for water. The revised
bottle test (1978) new product evaluation section is the basis for a toxicity
assay screening protocol which is in the validation process.
Algal assays are a sensitive test for evaluating the effects of heavy
metals and for predicting the future effects of toxics in aquatic systems
(Chiaudani and Vighi, 1978). Recognition of the application of algal assays
and the usefulness of their results for water quality management is increasing
throughout the world. The 1978 Algal Assay protocol (Miller, Greene and
Shiroyama, 1978) has been distributed throughout the United States and to more
than 40 foreign countries.
The purpose of this bibliography is to supplement the 1978 Algal Assay
protocol by compiling the available literature, published and unpublished, on
research that contained algal assays using Selenastrum. The papers cited in
this bibliography include studys of the effects of nutrients, toxicants,
complex wastes, and specific inorganic and organic compounds upon
Selenastrum's growth and physiology. The result is a collection of abstracts,
citations, and keywords describing 220 papers. This bibliography is designed
to be used as a public reference for managers of water quality programs and
scientists or educators with interests in research pertaining to Selenastrum
and algal assays.
A general review of the cited literature indicates that the standard
algal assay procedure was followed, but often with modifications to meet the
needs of each investigation. In order to compare and utilize the results of
similar studies, standardization of the algal assay procedure must be main-
tained. When deviations from the normal test procedures are deemed necessary,
IV
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the standard procedure should be performed simultaneously for control pur-
poses. The modified algal assay results can then be compared to those
achieved by following the recommended procedures and recognized improvements
in methodology can be used to improve the standard procedure. This, hope-
fully, will be a continuing process. Review of the literature also indicates
that a trend has been developing over the years towards a more consistent
practice of following standard algal assay procedures such as use of stand-
ards: growth media; growth conditions (i.e., light, temperature); inoculum
size; and, methods used to measure and assess growth. Utilization of a stand-
ardized algal assay procedure, using a selected universal organism, would
continue the advancements made towards building a comparable and highly appli-
cable data base for management of water quality.
This bibliography would not have been possible without the generous
efforts of those individuals who recognized the requests for literature per-
taining to Selenastrum and responded with reprints and/or citations of their
research papers. The kindness of the individuals and organizations that
granted us the privilege of reproducing copyrighted abstracts and summaries is
greatly appreciated. Without these the usefulness of the bibliography would
have been greatly reduced. The patience and continued efforts of Zilla
Toohill and Bonnie Byron in typing the bibliography are also greatly appre-
ciated.
A standard format was followed for the presentation of the literature.
Each paper is presented as: a complete citation with an associated accession
number; a list of keywords; and, an abstract or summary provided by the au-
thor(s) or the properly acknowledged source. An alphabetized keyword index
and author index follows the bibliographic list.
The format for the citations was derived from that followed by the jour-
nal Limnology and Oceanography (American Society of Limnology and Oceanog-
raphy) and the 4th Edition Council of Biology Editors Style Manual (Council of
Biology, Editors Inc. 1978). Abbreviations for the journals followed Biosis:
List o_f Serials (BioScience Information Service, 1977). Exceptions to the
basic format are:
1) The accession number was designed initially as a four-digit number.
As a result of papers being added after the first number assignments and
because we wished to maintain alphabetization of authors names, lower case
letters were added to the four-digit number, where affected.
2) Citations are provided with brief keyword lists for two papers
(0098-b, 0098-c) that were not retrievable. A citation, keyword list and an
abstract are provided for paper 0085-a, also not retrievable. Either the
translations of the papers' title or source were in error. An extensive
library search for these papers proved fruitless, but it was decided to in-
clude the citations. Hopefully, someone may recognize and correct the infor-
mation that has been provided.
3) A few of the papers were published in a foreign language. All are
provided with the translated title, source and abstract or summary. The
language in which the paper was published is indicated in the citation.
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4) Additions to the bibliography were not accepted after 1 April, 1979
so that the project could move toward publication.
5) It is requested that copies of papers using Selenastrum in algal
assays that are not included in this bibliography, and any future papers
discussing Selenastrum, be forwarded to Joseph C. Greene, Corvallis Environ-
mental Research Laboratory, Corvallis, Oregon 97330, to be included in a
future addendum.
REFERENCES
American Public Health Association, American Water Works Association, and
Water Pollution Control Federation. 1975. Standard methods for the
examination of water and wastewater, 14th edition. American Public
Health Association, Washington, D.C. 1193 p.
BioSciences Information Service. 1977. Biosis: List of serials. Bio-
Sciences Information Services, Philadelphia, Pennsylvania. 251 p.
Chiaudani, G. , and M. Vighi. 1978. The use of Selenastrum capricornutum
batch cultures in toxicity studies. Mitt. Int. Ver. Limnol. 21:316-329.
Council of Biology Editors, Inc. 1978. Council of Biology Editors style
manual. A guide for authors, editors, and publishers in the biological
sciences. 4th edition. Amer. Inst. Biol. Sci., Arlington, Virginia.
265. p.
Miller, W.E., J.C. Greene, and T. Shiroyama. 1978. The Selenastrum
capricornutum Printz Algal Assay Bottle Test: Experimental design,
application, and data interpretation protocol. U.S. Environmental Pro-
tection Agency, Corvallis, Oregon. EPA-600/9-78-018. 125 p.
U.S. Environmental Protection Agency. 1971. Algal Assay Procedure Bottle
Test. U.S. Environmental Protection Agency, Corvallis, Oregon. 82 p.
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TABLE OF CONTENTS
Page
Foreword iii
Preface iv
Bibliography 1
Keyword Index 144
Author Index 179
VI1
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1.0 Bibliography
In accordance with copyright regulations, reprints other than our own are
not available from the Corvallis Environmental Research Laboratory.
0001-a
Adam, V. D. , D. B. Porcella, and M. L. Cleave. 1978. The biological role of
specific compounds in aquatic ecosystems produced by oil shale develop-
ment, p. 73-74. In Utah Water Res. Lab. & Center Water Res. Res.,
Annual Report UWRL G-78/03.
bottle test, algal assay, oil shale, leachates, lake waters, algistatic, salt
loading, stimulation, algae, Selenastrum capricornutum, phytoplankton, Synedra
delicatissina, Scenedesmus quadricauda, diatoms.
The development of the soil shale industry will produce large quantities
of retorted shale during shale oil production. This retorted shale contains
high levels of salt substances and some organic compounds, and water passing
through it will leach these salts and possibly the organics into downstream
surface and groundwaters. This salt-laden water could have a significant
impact on the growth of freshwater algae if it enters the Colorado River
system. Since algae are the basis for the food chain in the river, any effect
on the algal productivity could affect the food supply for all higher life
forms in the river. Since the salts common in the retorted shale are also
common to the soil in the region, laboratory experiments that determine the
effects of increased salt loading on algae are also applicable in determining
the effects of irrigation practices and other water usage which causes evapo-
rative losses in the river system. (Abstract-Author).
0001-b
American type culture collection. 1975. Collections of algae and protozoa -
List of strains. ATCC, Rockville, Maryland. 9 p.
algae, cultures, protozoa, Selenastrum capricornutum ATCC #22662.
A list of strains of algae and protozoa available and the ATCC classifi-
cation number.
0001
Aronson, J. G. 1973. The effect of some common insecticides upon C-14 uptake
in phytoplankton. M.S. thesis, Univ. Nebraska, Lincoln. 96 p.
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carbon-14, insecticides, organochlorine compounds, algae, Anabaena flos-aquae,
Microcystis aeruginosa, Selenastrum capricornutum Printz, algal assay.
The acute effect of the twelve organochlorine compounds DDT, DDE, BHC,
methoxychlor, lindane, chlordane, aldrin, dieldrin, endrin, toxaphene, endo-
sulfan, and mi rex upon the carbon-14 uptake of i_n situ and it\ vitro phyto-
plankton was determined utilizing the C-14 technique of Steeman-Nielsen. The
IQ s11 u experiments were performed in Pawnee Reservoir, Lancaster County,
Nebraska. The ijn vitro experiments were performed using unialgal cultures of
Anabaena flos-aquae (Lyngb.) DeBrebisson, Microcystis aeruginosa Kutzing amend
Elenkin (Anacystis cyanea Drouet and Daily), and Selenastrum capricornutum
Printz. Initial concentrations of l.Oppm, O.lppm, O.Olppm, and l.Oppb of each
compound were used for each series of tests.
In situ C-14 uptake was reduced by all twelve chemicals. DDT, its decom-
position product DDE, and a close structural relative, methoxychlor, were most
effective in reducing ijn situ C-14 uptake. Ijn vitro results were variable.
While the C-14 uptake of Anabaena flos-aquae cultures was reduced by all
twelve chemicals, that of the Microcystis aeruginosa cultures was generally
enhanced. Cultures of Selenastrum capricornutum were relatively unaffected by
any of the chemicals. (Abstract-Author).
0002
Bartlett, L., F. W. Rabe, and W. H. Funk. 1974. Effects of copper, zinc, and
cadmium on Selenastrum capricornutum. Water Res. 8(3):179-185.
zinc, copper, cadmium, algae, Selenastrum capricornutum, algicidal, algi-
static, bottle test, rivers, algal assay.
The algicidal and algistatic effects of copper, zinc and cadmium on
Selenastrum capricornutum, a unicellular green algae were analyzed by using a
modification of the Algal Assay Procedure Bottle Test.
Algicidal concentrations of copper, zinc and cadmium were 0.30, 0.70, and
0.65 mg I-1. Treatment of Selenastrum with various concentrations of the
metals resulted in similar growth rates characterized by extended lag growth
phases.
Combinations of copper, zinc and cadmium were similar in toxicity to
equal concentrations of zinc. Combinations of copper and cadmium resulted in
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a greater growth rate than equal concentrations of copper suggesting that
cadmium inhibits copper toxicity.
Selenastrum was able to exist in waters from the upper South Fork and
North Fork of the Coeur d'Alene River where zinc and other metals were in low
concentrations. However, the algae was not able to tolerate zinc concentra-
tions greater than 0.5 mg I-1 from waters of other parts of the drainage.
These observations were consistent with laboratory findings where 0.7 mg I-1
zinc was algicidal and 0.1 mg I-1 inhibited the growth of Selenastrum. (Ab-
stract-Author).
0003
Behnken, D. W. , and D. G. Watts. 1972. Bayesian estimation and design of
experiments for growth rates when sampling from the Poisson distribution.
Biometrics 28(4):999-1009.
data interpretation, experimental design, Poisson distribution, algae, Sele-
nastrum capricornutum, growth rates, statistical methods, algal assay.
Data on algal growth rates were used to investigate the problem of esti-
mating the growth rate parameter, B, for a process with an expected value X.
P+-. i
equal to aep ^. This equation produces data from a Poisson distribution P of
lambda A. at time t, (i equals 1,2, n). The posterior distribution for p
is derived, and an approach to designing experiments in this situation is sug-
gested. (Abstract-Little-Battelle WRSIC).
0003-a
Bentley. R. E. , J. W. Dean, S. J. Ells, T. A. Hollister, G. A. LeBlanc, S.
Sauter, and B. H. Sleight. 1977. Laboratory evaluation of the toxicity
of cyclotrimethylene trinitramine (RDX) to aquatic organisms. Final
report, E G & G Bionomics, Wareham, Mass. DAMD-17-74-C-4101. 86 p.
toxicity, algistatic, cyclotrimethylene trinitramine, RDX, algal assay, bottle
test, chlorophyll a, algae, Microcystis aeruginosa, Selenastrum capricornutum,
Anabaena flos-aquae, diatoms, Navicula pelliculosa, fish, water quality.
The toxicity of nitroglycerine to a wide variety of aquatic organisms
representing several different trophic levels in aquatic ecosystems was stud-
ied. Results of static acute toxicity tests indicate that the acute LC50
values are greater than 3 mg/1 RDX. There was an apparent lack of bioaccumu-
lation in edible or nonedible tissues or organs in all species tested. Ef-
-------
fects were observed on growth at 5.8 mg/1 RDX during egg and fry studies, on
survival at 4.9-63 mg/1 during chronic exposure of fathead minnows, and on
number of young produced per parthenogenetic female at concentrations > 4.8
mg/1. Applying an application factor of 0.1 to the lower limit of observed
acute toxicity values, (3.6 mg/1, 96-hour LC50 for bluegill at pH 6.0, 20°C) a
water quality criterion of 0.35 mg/1 RDX is proposed for the protection of
freshwater aquatic life with an adequate margin of safety. (Abstract-Author).
0003-b
Bentley, R. E. , G. A. LeBlanc, T. A. Hollister, and B. H. Sleight III. 1977.
Acute toxicity of 1,3,5,7-tetranitro-octahydro-l,3,5,7-tetrazocine (HMX)
to aquatic organisms. Final report, E G & G, Bionomics, Wareham, Mass.,
DAMD-17-74-C-4101. 23 p.
toxicity, HMX, 1,3,5,7-tetranitro-octahydro-l,3,5,7-tetrazocine, water qual-
ity, phytoplankton, fish, algae, Microcystis aeruginosa, Anabaena flos-aquae,
Selenastrum capricornutum, diatoms, Navicula pelliculosa, algal assay, bottle
test.
The acute toxicity of 1,3,5,7-tetranitro-octahydro-l,3,5,7-tetrazocine
(HMX) was studied utilizing aquatic organisms representing several different
trophic levels in aquatic ecosystems. Generally, no adverse effects of expo-
sure to 32 mg/1 HMX were observed among any of the algae, fish, or inverte-
brate species tested. The 7-day old fry of the fathead minnow were the only
life stage or species acutely affected. Based on an application factor of
0.05 and a 96-hour LC50 for the most sensitive aquatic organism (7-day old fry
of the fathead minnow) tested (15 mg/1), a water quality criterion of 0.75
mg/1 is proposed for the protection of freshwater aquatic life with an ade-
quate margin of safety. (Abstract-Author).
0003-c
Bentley, R. E. , J. W. Dean, S. J. Ells, G. A. LeBlanc, S. Sauter, K. S.
Buxton, and B. H. Sleight, III. 1978. Laboratory evaluation of the
toxicity of nitroglycerine to aquatic organisms. Final report. E G & G,
Bionomics, Wareham, Mass. DAMD-17-74-C-4101. 82 p.
toxicity, nitroglycerine, water quality, algistatic, algal assay, bottle test,
chlorophyll a, algae, Microcystis aeruginosa, Anabaena flos-aquae, Selenastrum
capricornutum, diatoms, Navicula pelliculosa.
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The toxicity of nitroglycerine to a wide variety of aquatic organisms
representing several different trophic levels in aquatic ecosystems was stud-
ied. The results of static acute toxicity tests indicate that the nominal
acute LC50 values are generally greater than 1 mg/1 nitroglycerine. A pilot
bioaccumulation study with C-14-nitroglycerine showed that there was no appre-
ciable accumulation of C-14-nitroglycerine in the edible tissues of four fish
species. The estimated application factor for fathead minnow over one com-
plete life cycle is >0.01<0.02 and >0.03<0.07 for eggs and fry of the fathead
minnow and channel catfish, >0.2<0.4 for daphnids and >0.3<0.6 for midges.
Applying the lower limit of all estimates of the application factor (0.03) to
the lower limit of the observed acute toxicity values for freshwater organisms
(1.28 mg/1, 96-hour LC50 for bluegill at pH 6.0), we propose a water quality
criterion of 0.01 mg/1 nitroglycerine. (Abstract-Author).
0003-d
Bentley, R. E. , J. W. Dean, T. A. Hollister, G. A. LeBlanc, S. Sauter, B. H.
Sleight III, and W. G. Wilson. 1978. Laboratory evaluation of the
toxicity of elemental phosphorus (P4) to aquatic organisms. Final Re-
port, E G & G, Bionomics, Wareham, Mass. DAMD-17-74-C-4101. 92 p.
toxicity, phosphorus, algistatic, algal assay, bottle test, chlorophyll a,
algae, Microcystis aeruginosa, Anabaena flos-aquae, Selenastrum capricornutum
diatoms.
The toxicity of elemental phosphorus to a wide variety of aquatic organ-
isms was studied. Results of static acute toxicity tests indicate that the
acute LC50 values varied over several orders of magnitude with an observed
range of 6 - >560 ug/1 of elemental phosphorus. Chronic exposure of fathead
minnows for 150 days to concentrations ranging from 1.5 - 3.4 |jg/l signifi-
cantly reduced growth. In addition, survival was affected in concentrations
>1.5 ug/1, while eggs spawned by fish exposed to the low concentrations of
0.71 and 0.40 (jg/1 failed to hatch when incubated. Although bioaccumulation
of elemental phosphorus was low (ca 200X), the cumulative mortality pattern
observed suggests continuing damage to target organs. Based on the available
data and the use of a safety factor of 0.1, an ambient water quality criterion
for elemental phosphorus of 0.04 ug/1 is recommended (Abstract-Author).
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0003-e
Bilcea, R. 1975. Contributions to the knowledge of the influence of nutri-
tion on the growth and development of the alga Selenastrum gracile
Reinch. Rev. Romaine de Biol. 20(3):185-191.
algal assay, algae, N:P ratio, Selenastrum gracile Reinsch, soil extraction.
The physiological characters of S. gracile grown in various nutritive
media with a small soil extract supplement in various concentrations were
studied. The alga thrived on concentrated nutritive media with a N:P ratio of
5.5:1. The soil extract at a concentration of 1% stimulated the multiplica-
tion of the alga in all the nutritive media. (Abstract-Biological Abstracts,
Inc.).
0004
Bishop, N. I., M. Frick, and L. W. Jones. 1977. Photohydrogen production in
green algae: Water serves as the primary substrate for hydrogen and
oxygen production, p. 3-22. In A. Mitsui, S. Miyachi, A. San Pietro, and
S. Tamura [eds.], Biological solar energy conversion, Academic Press, New
York.
DCMU, CCCP, photohydrogen, algae, Selenastrum sp. , Ankistrodesmus falcatus,
Ankistrodesmus braunii, Scenedesmus, Chlorella fusca, Chlorella pyrenoidosa
(211-8k), Chlorella kesslerii, Kirchneriella lunaris, Coelastrum proboscideum,
Ulothrix acuminata, Scenedesmus obliquus, algal assay.
Photohydrogen evolution was surveyed in a number of species of algae.
The following ones had rates of H2 evolution comparable to Scenedesmus
obliquus: Selenastrum sp. , Ankistrodesmus falcatus, Ankistrodesmus braunii,
Scenedesmus sp. , Kirchneriella lunaris, Coelastrum proboscideum, Ulothrix
acuminata, Chlorella fusca, Chlorella pyrenoidosa (211-8K), and Chlorella
kesslerii. Maximum rates for autotrophic and heterotrophic: Scenedesmus
approached 2 and 10 (jl H2/ul cells/hr respectively, 50-90% inhibition of H2
production by DCMU was uniformily observed in all of the above species and in
manganese deficient Scenedesmus.
Re-examination of photohydrogen production by a series of Scenedesmus
mutants blocked in PS-II revealed a nearly complete loss of photohydrogen
production. CCCP stimulated the initial low rates but only to about 10% of
wild type. We conclude that hydrogen evolution in the species examined re-
-------
quires both photosystems for maximal activity; whether water photolysis is
essential for this reaction remains an open question. (Abstract-Author).
0005
Bowman, R. H. 1974. The control of pH by carbon dioxide, TRIS (Hydroxymethyl
aminomethane), and other buffers on the Algal Assay Bottle Test. M.S.
thesis, Utah State Univ., Logan. 100 p.
algal assay, bottle test, pH, carbon dioxide, TRIS, buffers, algae, Sele-
nastrum capricornutum, reservoirs, water hardness.
The purpose of this paper is to discuss the development of a convenient
method for using the Environmental Protection Agency's Algal Assay Procedure
Bottle Test in hard waters and to demonstrate the usefulness of the technique
in assays of Hyrum Reservoir water samples.
This thesis also deals with the problem of using the bottle test in hard
water. This problem is that when the pH rises above 9.0, a chemical precipi-
tation is observed on the bottom of the flasks and all algal growth stops.
The first step in developing a convenient method for using the assay in hard
water was to determine a pH range at which the test alga (Selenastrum capri-
cornutum) would grow but precipitation would not occur- Once the pH range was
determined, it was necessary to develop a suitable method for controlling the
pH in that range.
Two methods for controlling pH were evaluated in this experiment: Tris
(hydroxymethyl aminomethane) buffer and C02 addition. Other buffers were
evaluated in terms of pH control within a specific range, availability and
cost, ease of addition and effect of buffer on algal growth. (Abstract-
Author).
0006
Brezonik, P. L. , F. X. Browne, and J. L. Fox. 1975. Application of ATP to
plankton biomass and bioassay studies. Water Res. 9(2):155-162.
ATP, chlorophyll a, algal assay, DNA, luminescence, algae, Anabaena flos-
aquae, Microcystis aeruginosa, Selenastrum capricornutum, phytoplankton,
Chlorella sp.
The use of adenosine triphosphate (ATP) analysis for freshwater algal and
plankton populations was evaluated as a measure of biomass and as a bioassay
-------
response parameter. ATP analysis was performed using the firefly luminescence
procedure. In short term laboratory studies, ATP levels in cultured algae and
lake plankton correlate well with other standard biomass parameters, including
chlorophyll a and dry weight. Algal ATP responded rapidly to mercury addition
and pH changes, indicating its usefulness as a measurement of toxicity. The
rapid response of ATP following nutrient additions to starved algal cultures
suggests ATP may be useful as a tool in nutrient bioassay studies. (Abstract-
Author).
0007
Brown, E. J. , and R. F. Harris. 1978. Kinetics of algal transient phosphate
uptake and the cell quota concept. Limnol. Oceanogr. 23(1):35-40.
phosphate, phosphorus, limiting nutrients, uptake, cell quota, growth kinet-
ics, bottle test, algae, Selenastrum capricornutum, Nostoc sp., algal assay.
Dissolved inorganic orthophosphate (Px) uptake rates (v) were determined
as a function of total cell phosphorus content (Q) in P± perturbed axenic
batch cultures of Selenastrum capricornutum and Nostoc sp. The results showed
an inverse relationship between v and Q for both organisms. These findings
are used to relate the cell quota concept of nutrient limited, steady state
microbial growth kinetics to nutrient uptake kinetics observed in perturbed
batch cultures. (Abstract-Author).
0008
Brown, E. J. , R. F. Harris, and J. K. Koonce. 1978. Kinetics of phosphate
uptake and aquatic microorganisms: Deviations from a simple Michaelis-
Menten equation. Limnol. Oceanogr. 23(l):26-34.
kinetics, phosphate, 32P, uptake, Michaelis-Menten equation, bottle test,
algae, Selenastrum capricornutum, phytoplankton, lake waters, algal assay.
The uptake kinetics of dissolved inorganic phosphate (31Pi) by lake water
microorganisms and a green alga (Selenastrum capricornutum) were determined as
a function of uptake rates of radiophosphate (32Pi) at added 31P! levels >0.
The data did not follow a simple Michaelis-Menten equation. (Abstract-
Author).
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0008-a
Brown, E. J. , and D. K. Button. 1979. Phosphate-1imi ted growth kinetics of
Selenastrum capricornutum (Chlorophyceae). J. Phycol. In press.
phosphate, limiting nutrients, kinetics, growth rates, continuous flow, nutri-
ents, algal assay, algae, Selenastrum capricornutum Printz.
Most theoretical studies of phytoplankton growth in aquatic environments
assume that relative nutrient utilization abilities regulate species composi-
tion. The steady-state phosphate-limited growth kinetics of Selenastrum
capricornutum Printz were examined using continuous cultures to characterize
the green alga's ability to compete for orthophosphate (Pi) when Pi limits
growth. The maximal specific growth rate for Selenastrum at 20°C was 1.20
(day)-1, and the concentration where half maximal growth rate occurs was 40
nanomolar Pi. There was an apparent threshold of 10 nanomolar Pi. Cell
yields varied inversely with growth rate; thus ability to utilize Pi could not
be characterized in terms of the Monod half-saturation constant and maximal
growth rates. Instead, we computed the Pi affinity from steady-state flux vs.
external Pi concentrations. This affinity was 2.8 liter (milligrams dry
weight-day)-1 for Selenastrum. Kinetic evidence from this study suggests that
Selenastrum will not be growth competitive with some other common aquatic
heterotrophs and autotrophs when Pi limits microbial growth in lakes. (Ab-
stract-Author).
0009
Cain, J. R. , and F. R. Trainor. 1973. A bioassay compromise. Phycologia
12(3/4):227-232.
algal assay, water quality, nutrients, growth rates, bioindicators, rivers,
wastewater, algae, Selenastrum capricornutum.
Selenastrum capricornutum was used as a test organism for assaying the
nutrient condition of natural waters. Using an actively growing culture,
inoculated, sterile-filtered water was incubated at 22° C in continuous fluor-
escent illumination. Organisms were grown in tubes on a roller tube rotator
and transferred daily, re-establishing a population of 4 x 105 cells each day.
Averages of 5-day runs showed no evidence for nutrient carry-over when 1 +
doublings/day were recorded; carry-over usually was not apparent on days 3-5
when nutrient poor waters were examined.
-------
We examined sites above and below a sewage treatment plant, in different
rivers and during fall, winter and spring. The assay, as measured in doub-
lings of Selenastrum/day, shows clear differences in the carrying capacity of
the rivers at these sites. (Abstract Only-Author).
0009-a
Camp, F. A., J. M. Dolan III, and% A. C. Hendricks. 1974. Algal bioassay
studies of the effects of bio-degradation on the toxicity of a non-ionic
surfactant to Selenastrum capricornutum, Chlorophyceae. ASB Bull.
21(2):45.
wastewater, biodegradation, algal assay, toxicity, algae, Selenastrum capri-
cornutum, bottle test.
A mixture of a nonionic surfactant and synthetic sewage was introduced
into bench scale activated sludge units. Retention times of 4 and 12 hours
were used to vary the efficiency of biodegradation of the surfactant. Both
control and experimental units were operated for each retention time. E.P.A.
algal bioacivjy techniques were used to evaluate the toxicity of the effluents
to the test organism Selenastrum capricornutum. Growth was monitored regu-
larly by reading absorbance with a double-beam spectophotometer. Results were
statistically significant (a=0.05) in showing that experimental effluents
supported essentially the same levels of growth as the control effluents.
This was found for both the 4 hour and 12 hour effluents, i.e. no toxic effect
was observed. (Abstract Only-Author).
0010
Chiaudani, G. , and M. -Vighi. 1974. The N:P ratio and tests with Selenastrum
to predict eutrophication in lakes. Water Res. 8(12):1063-1069.
nitrate, phosphate, algae, Selenastrum capricornutum, eutrophication, lake
waters, limiting nutrients, N:P ratios, algal assay.
During the spring overturn, at the time of maximum potential phytoplank-
ton growth, 26 Italian lakes were sampled to carry out enrichment experiments
in which single and mixed treatments of nitrate and phosphate were applied to
laboratory cultures of Selenastrum capricornutum. The method, in conjunction
with chemical analysis, can be used to classify the lakes and to evaluate the
limiting values of each nutrient. It was possible to correlate the phosphorus
10
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concentration to the maximal primary production and obtain threshold levels
for phosphorus in predicting eutrophication trends in lakes. (Abstract-
Author).
0011
Chiaudani, G. , and M. Vighi. 1975. Dynamic of nutrient limitation in six
small lakes. Verh. Int. Ver. Limnol. 19(2):1319-1324.
bottle test, algal assay, algae, Selenastrum capricornutum, limiting nutri-
ents, lake waters, eutrophication, phosphorus, nitrogen.
Laboratory bioassays of samples which utilized Selenastrum capricornutum
(Printz) as the test organism, were carried out monthly in 1972-73 on six
small subalpine lakes (the Brianza Lakes) in northern Italy near the southern
part of Lake Como. The bioassays were aimed at establishing experimentally
which of the essential mineral elements must be removed to restrain or cut
down excessive algal growth in lake waters. Measurements were made of ortho-
phosphate phosphorus, total soluble phosphorus, ammonia nitrogen, total inor-
ganic carbon (computed from total alkalinity, acidity and temperature) and
three measured nitrogen oxidation forms. Morphological characteristics were
examined in the lakes, and a stepwise multiple regression analysis was run to
evaluate the statistical relationship between S. capricornutum growth as a
dependent variable and different chemical parameters as independent variables.
Three conclusions were drawn from this analysis: (1) The Molybdenum Blue
Method, as confirmed by biological assays, measures the phosphorus available
for algal growth; (2) Orthophosphate phosphorus appears to be the key factor
for phytoplankton growth in all six lakes, followed by nitrogen as the factor
of second-ranked importance and as verified by the stepwise multiple regres-
sion analysis; (3) The evaluation of the nitrogen:phosphorus ratio in lake
waters during winter overturn can reasonably permit forecasts of the dynamics
of nutrient limitation all through the year. (Abstract-Harris-Wisconsin
WRSIC).
0012
Chiaudani, G., and M. Vighi. 1976. Comparison of different techniques for
detecting limiting or surplus nitrogen in batch cultures of Selenastrum
capricornutum. Water Res. 10(8): 725-729.
11
-------
nitrogen, limiting nutrients, bottle test, N:P ratios, algae, Selenastrum
capricornutum, algal assay.
Several metabolic responses of Selenastrum capricornutum grown at differ-
ent initial N:P ratios were studied in order to determine the degree of nitro-
gen deficiency when alga enters and leaves nitrogen limitation in batch cul-
tures. The results show a good agreement among the tests and good sensitivity
and accuracy for all the used techniques. (Abstract-Author).
0013
Chiaudani, G., and M. Vighi. 1978. The use of Selenastrum capricornutum
batch cultures in toxicity studies. Mitt. Int. Ver. Limnol. 21:316-329.
bottle test, toxicity, heavy metals, copper, zinc, cadmium, nickel, chromium,
cobalt, lake waters, algae, Selenastrum capricornutum, algal assay.
Selenastrum capricornutum batch cultures are extensively applied to the
assessment of algal growth potential and to identify limiting factors.
In the present paper, some results of the use of a standard assay pro-
cedure for the identification of toxic effects in natural waters, are pre-
sented. The presence of toxic effect is evaluated on the basis of the compar-
ison between observed growth and the growth predicted from the concentration
of the limiting nutrient.
Also, some data relative to the toxicity of selected heavy metals as EC50
96-hr on Selenastrum capricornutum grown in AAP completely inorganic medium
and in medium with a chelating agent are given.
Finally, with the aim to set up water quality criteria for phytoplankton
some results are given relative to the effects of some heavy metals added to
natural waters taken from largely different environments. (Abstract-Author).
0013-a
Chillingsworth, M. B. 1974. The toxicity of aminoanthraquinone dyes to fish
and algae. Chap. 5. Jj} Dyes in the environment - Reports on selected
dyes and their effects, v. II, Amer. Dye Manuf. Inst., Inc. , New York.
dyes, fish, Pimephales promelas, algal assay, bottle test, algae, Selenastrum
capricornutum, toxicity.
12
-------
Twelve aminoanthraquinone dyes—Disperse Violet 1, Disperse Blue 3,
Solvent Blue II, Solvent Blue 14, Solvent Green 3, Acid Green 25, Acid Violet
34, Acid Blue 25, Disperse Blue 7, Acid Blue 45, Basic Blue 21, and Disperse
Red 60—and three anthraquinone compounds--anthraquinone, sodium anthraquinone
beta sulfonates, and 1-anthraquinone sulfanilic acid sodium salt—were studied
for their toxicity to the fathead minnow, Pimephales promelas, using the acute
static bioassay procedure and to the green algae, Selenastrum capricornutum,
using the algal assay bottle test. The anthraquinone compounds exhibited no
toxic effects to the fish in 96 hours at concentrations of 1 mg/1 and 10 mg/1.
Symmetrical dyes, Acid Blue 45 and Acid Violet 34, were also nontoxic. The
strongly cationic dye, Basic Blue 21, is highly toxic to both fish and algae.
The remainder of the dyes showed varying degrees of toxicity related to the
degree of ionization. Within this group the pattern of response by fish and
algae differs, possibly indicating different toxic mechanisms. Reax 83-A,
used to disperse the solvent and disperse dyes tested, or impurities in the
dyes may contribute to the toxicity observed. Further investigation of an-
thraquinone dyes is recommended to elucidate the toxic modes and to enable
better predictions of toxicity. (Abstract-Author).
0013-b
Christensen, E. R. , and J. Scherfig. 1979. Effects of manganese, copper and
lead on Selenastrum capricornutum and Chlorella stigmatophora. Water
Res. 13(l):79-92.
algal assay, algae, bottle test, Chlorella stigmatophora, copper, inhibition,
lead, manganese, media, Selenastrum capricornutum, toxicity, wastewater, heavy
metals.
Algal assays were conducted to observe the responses of Selenastrum
capricornutum and Chlorella stigmatophora to the metals manganese, copper and
lead added singly or in combination to both artificial media and natural
waters. A 50% reduction in the total algal cell volume of Selenastrum in
standard algal medium (SAAM) occurred by the addition of either 3.1 mg manga-
nese I-1, 85 ug copper I-1, or 140 u lead I-1. For Chlorella grown in 28 ppt
artificial seawater plus full SAAM nutrients, the corresponding numbers were
50 mg manganese I-1, 70 pg copper I-1, or 700 \jg lead I-1. Of the metals
tested, only lead was found in higher concentrations in urban and agricultural
runoff than at least one of these levels.
13
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Theoretical considerations show that a product model is a more reasonable
reference for no-interaction than an additive model for experiments in which
toxic metals are added in combination. Combination experiments, in which any
combination of the three metals act at the same time indicate (1) synergism
between manganese and copper (2) antagonism between manganese and lead and (3)
antagonism between copper and lead. As to the manganese-lead interaction, it
is shown that the lead-induced inhibition of cell division is offset, par-
tially for Selenastrum and completely for Chlorella by the addition of mangan-
ese. (Abstract-Author).
0014
Claesson, A. 1973. Algal assay procedure: Minitest with lake water (prelim-
inary report), p. 35-40. In. Algal assays in water pollution research,
Proc. Nordic Symp., 25-26 October 1972, Oslo, Norway. NORDFORSK, Secre-
tariat of Environmental Sciences.
lake waters, Minitest, algae, Selenastrum capricornutum, Ankistrodesmus
falcatus, Chlorella homosphaera, Euglena gracilis, Scenedesmus quadricauda,
AGP, limiting nutrients, algal assay, bottle test.
The report describes the use of the Minitest (National Swedish Environ-
ment Protection Board Algal Assay) to test the growth potential of lake waters
and to study the limiting nutrients. The algal species used was Selenastrum
capricornutum. Results are briefly discussed and growth curves are provided.
The author concludes that the algal assay procedure can be successfully con-
ducted with small volumes of test waters, thus the name "Minitest". (Ab-
stract-Lei schman-EPA).
0015
Claesson, A. 1975. Algal assay procedures at some Nordic laboratories - a
comparison. Vatten 31 (4): 333-338.
bottle test, algae, Selenastrum capricornutum, Chlorella sp. , Chlorella homo-
sphaera, Phaeodactylum tricornutum. Ankistrodesmus falcatus, Oscillatoria
agardii, AGP, limiting nutrient, algal assay, diatoms.
A comparison between the different algal assay procedures used at some
Nordic laboratories gave the following results:
14
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1. A comparison between the precision of the electronic particle counters
indicated the necessity of corrections. The following intercalibrations
then gave good agreements between the countings.
2. The maximal biomass produced varied a great deal between the labora-
tories. The same experience has been obtained from similar American
investigations.
3. The different laboratories determined with good agreement P or N as the
most limiting nutrients.
4. The growth curves for Selenastrum in standard medium showed good agree-
ment between three of four participating laboratories when maximal yield
(measured as cell volume) was determined, but the algal cells were of
different size (mean cell volume). (Summary-Author).
0016
Claesson, A. 1978. Research on recovery of polluted lakes: Algal growth
potential and the availability of limiting nutrients. Abstr. Uppsala
Diss, ACTA Univ. Upsaliensis No. 461, Uppsala, Sweden. 27 p.
Minitest, lake waters, AGP, algal assay, limiting nutrients, algae, Sele-
nastrum capricornutum, wastewater, N:P ratios.
In order to elucidate the effects of advanced wastewater treatment for P
removal and sewage diversion, the National Swedish Environment Protection
Board started a long-term program in 1972 for analyzing the conditions in a
number of wastewater receiving lakes. This thesis summarizes some algal assay
results from some of these polluted lakes mainly concerning algal growth
potential (AGP), growth-limiting nutrients and availability of algal nutri-
ents. As a result of the present investigation a few general remarks can be
made.
- In lake water a total-N: total-P (TN:TP) ratio of > 17 and an inorganic-
N: inorganic-P (IN: IP) ratio of > 12 can be used as critical values for
P-limited Selenastrum growth and for N-limited growth a TN:TP ratio of £
10 and an IN: IP ratio of < 5. These values are suggested for use only
when no algal assay data are available.
- A decrease in the nutrient content of a lakewater can result in decreased
lake chlorophyll a content or in decreased AGP or in both simultaneously.
15
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- The potential algal biomass should be considered when describing the
trophic status of a water body e.g. as "total chlorophyll a".
- The degree of nutrient- availability for algal growth of a water sample
can be estimated by algal assays.
- The amount of available algal nutrients ought to be considered in nutri-
ent budget calculations. (Abstract-Author).
0016-a
Claesson, A. 1978. Variation in cell composition and utilization of N and P
for growth of Selenastrum capricornutum. No. II, Publ. in Abstr. Uppsala
Diss. ACTA Univ. Upsaliensis No. 461, Uppsala, Sweden. 20 p.
nutrients, nitrogen, phosphorus, chlorophyll a, growth kinetics, N:P ratio,
algal assay, algae, Selenastrum capricornutum.
Yield coefficients (mm3 cell volume/mg nutrient element) for the green
alga Selenastrum capricornutum were determined with N03-N (YN = 134), NH4-N
(YN = 111) and P04-P (Yp = 2573).
Cell material from different phases in a growth curve was analyzed for
major elements and chlorophyll a content. The ranges in percent of the dry
weight were 50-80%, 4.4-8.8%, 0.24-3.0%, and 0.20-3.23% for C, N, P and chlor-
ophyll a respectively. The dry weight varied correspondingly 2.3 times. A
N/P-quotient of about 10 (by weight) was found to be a critical level for
Selenastrum when these elements were the most limiting nutrients for growth.
31 different organic N-compounds were tested as single N sources in growth
experiments with Selenastrum. At least 19 of them could be used - to varying
extents - for algal growth. (Abstract-Author).
0017
Claesson, A., and A. Forsberg. 1978. Algal assay procedure with one or five
species: Minitest. Mitt. Int. Ver. Limnol. 21:21-30.
algae, Selenastrum capricornutum, Ankistrodesmus falcatus, Chlorella homo-
sphaera, Euglena gracilis, Scenedesmus quadricauda, AGP, limiting nutrients,
lake waters, toxicity, wastewater, bottle test, Minitest, algal assay.
Two algal assay methods (batch), one with Selenastrum capricornutum and
one with this species together with Ankistrodesmus falcatus, Chlorella homo-
sphaera, Euglena gracilis, and Scenedesmus quadricauda mixed on equal cell
16
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volume basis, are described and compared. Selenastrum capricornutum is rou-
tinely used for determining the maximum algal growth potential (AGP) and the
limiting nutrients by enrichment experiments in lake water. The mixed algal
assay method is used to estimate AGP-values and to indicate toxic effects in
wastewater.
Different ways to prepare the algae before the assays are described and
discussed.
The tests are performed in small plastic tubes containing a culture
volume of 2.5 ml.
Different conditions influencing the growth of the algae were studied.
Examples of results from some treatment plants and some different lakes
using the two algal assay procedures are given, together with a comparison of
the two methods. (Summary-Author).
0017-a
Claesson, A., and S.-O. Ryding. 1977. Nitrogen-a growth limiting nutrient in
eutrophic lakes. Prog. Water Tech. 8(4/5):291-299.
algal assay, nitrogen, chlorophyll a, lake waters, N:P ratio, limiting nutri-
ents, AGP, algae, Selenastrum capricornutum.
Water chemistry and algal assays in surface water (0-2 m) are presented
from five eutrophic lakes, all recipients for sewage effluent. The lakes were
studied intensively during May-October 1973.
Some common features in water chemical aspects were: a decline in inor-
ganic-N during summer, P04-P always in excess, and a high chlorophyll a con-
tent throughout the period. The more productive lakes, however, exhibited a
high N supply even during intensive algal growth. The total-N:total-P ratio
was very low for most of the test period giving N limited algal growth with
the test alga Selenastrum capricornutum. A N:P ratio of 15:1 in the lake
water seemed to be a critical value concerning N- or P-limitation for this
test alga. The sum of lake- and algal assay chlorophyll was used to indicate
the algal growth potential (AGP) in lakes, and is proposed as a simple type of
"trophic index".
The lakes presented have been chosen to illustrate the variability
amongst lakes where N is the most limiting nutrient. In the less productive
ones N probably influenced the production level to a larger extent than in the
17
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more eutrophic ones, where other factors such as light, C-deficiency, or
zooplankton predation mainly limited phytoplankton growth. (Summary-Author).
0018
Clesceri, N. L. , G. C. McDonald, I. S. Kumar, and W. J. Green. 1973. Organic
nutrient factors affecting algal growths. Ecol. Res. Series. EPA-660/3-
73-003. 302 p.
wastewater, lake waters, organics, bottle test, algae, Microcystis aeruginosa,
Anabaena flos-aquae, Selenastrum capricornutum, algal assay.
Effects of wastewater organic fractions on the growth rate of Selenastrum
capricornutum and Anabaena flos-aquae were investigated. Effluent from a
conventional activated sludge facility was membrane filtered, freeze-dried,
and gel fractionated. Apparent molecular weights (AMW) were assigned to the
appropriate fractions. These and organic carbon data showed 69% of the efflu-
ent organics had an AMW less than 700.
Absorbancies and regression analyses within algal exponential growth
phases demonstrated the control growth rate for Selenastrum was 0.43 and for
Anabaena was 0.34. Selenastrum growth rates were monitored using Lake George
water as the diluent for the media employed. An inhibition in growth oc-
curred. Halving the nitrogen concentration in modified Gorham's had no sig-
nificant effect on growth rate.
In concentrating organics from natural water (Lake George and Saratoga
Lake), raw sewage, and sewage effluent, thin film evaporation was preferred
when using natural waters whereas freeze-drying was advantageous when working
with sewage samples. Also, the soluble organic component in municipal waste-
water was characterized and the effect of chemical-physical treatment on it
has been shown. (Abstract-Author).
0019
Condit, R. J. 1972. Phosphorus and algal growth in the Spokane River.
Northwest Sci. 46(3):177-189.
phosphorus, algae, Selenastrum capricornutum, phytoplankton, limiting nutri-
ents, rivers, bottle test, nitrogen, manganese, algal assay.
The green algal form, Selenastrum capricornutum, proved to be an adequate
and responsive test organism for natural water bioassays when properly accli-
18
-------
mated. The growth parameters obtained from control flasks indicate that
during the test periods, the total productivity and algal growth rates in the
natural waters increase as the river progresses through the city. During the
spring, high runoff provided adequate nutrients to promote algal growth in
bloom proportions. The large biomass reduced P concentrations to limiting
levels in the river. The especially high algal production in the lower river
was due in part to the added P made by the wastewater treatment plant above
the Bowl and Pitcher Bridge. Dissolved orthophosphate levels at this station
were sufficiently high to cause inhibition to algal growth rates when addi-
tional P was introduced in the bioassay. The August bioassay indicated that
productivity was governed by factors other than limiting concentrations of P.
Nitrogen was found to be limiting in Porcupine Bay while Mn appeared to be a
limiting factor in Sullivan Road waters. (Abstract-Biological Abstracts,
Inc.).
0020
Cowen, W. F. , and G. F. Lee. 1976. Phosphorus availability in particulate
materials transported by urban runoff. J. Water Pollut. Control Fed.
48(3)580-591.
nutrients, phosphorus, urban runoff, algal assay, Selenastrum capricornutum,
algae.
Samples of urban runoff were analyzed for dissolved reactive phosphorus,
total soluble phosphorus, particulate phosphorus, and total phosphorus.
Particles were isolated by membrane filtration and extracted with acid, base,
and anion exchange resins, and the inorganic phosphorus in the extracts were
compared with the particulate phosphorus extracted. Ranges of group mean
values for various urban land uses were 33 to 46, 22 to 27, and 13 to 17
percent of particulate phosphorus for acid, base, and resin extractions,
respectively. Bioassays with Selenastrum capricornutum showed a range of 8 to
55 percent of particulate phosphorus available to the algae. Long-term aero-
bic incubations of unfiltered runoff with resin generally showed the same
percentage of particulate phosphorus extractable by resin as did short-term
extraction. At most 30 percent of the particulate phosphorus should be con-
sidered available to algae in the receiving water. (Abstract-JWPCF).
19
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0020-a
Cowen, W. F. , and G. F- Lee. 1976. Algal nutrient availability and limi-
tation in Lake Ontario during IFYGL, Part I. Available phosphorus in
urban runoff and Lake Ontario tributary water. U. S. Environmental
Protection Agency, Duluth, Minn., Ecological Research Series, EPA-600/3-
76-094a. 218 p.
urban runoff, rivers, nutrients, phosphorus, algal assay, bottle test, algae,
Selenastrum capricornutum.
Urban runoff, precipitation, and river samples from Madison, Wisconsin
and New York state were analyzed for various phosphorus forms and the portion
of each phosphorus form available for algal use. Total phosphorus, soluble
phosphorus, particulate phosphorus, and soluble reactive forms were measured.
In addition, acid extractable, base extractable, and anion exchange resin
extractable inorganic phosphorus was determined on the particulate fractions.
Algal assay procedures were used to assess portions of the various phos-
phorus fractions available for Selenastrum capricornutum growth. Availability
of particulate phosphorus in urban runoff from Madison, Wisconsin was highly
variable ranging from 8 to 55 percent. Genesee River basin urban runoff had
from less than 1 to 24 percent of its particulate phosphorus available.
Particulate phosphorus from the Niagara, Genesee, Oswego, and Black Rivers
showed only 6 percent or less available to this alga. Autoclaving the samples
increased the amount of particulate phosphorus available. Precipitation
samples usually showed less than 9 percent of the total phosphorus available
to Selenastrum capricornutum. Total phosphorus available for algal growth
from New York tributaries was highly variable.
About 39, 24, and 15 percent of particulate phosphorus in urban runoff
from Madison could be extracted by acid, bases, and anion exchange. Results
from urban areas in the Genesee River basin in New York were similar. Resin
extractions in long-term aerobic dark incubations produced results similar to
short-term tests, indicating that physical and chemical rather than microbial
mineralization processes were probably the key factors regulating the release
of inorganic P from the runoff particules to the solution phase. (Abstract-
Author).
20
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0020-b
Cowen, W. F. , K. Sirisinha, and G. F. Lee. 1977. Algal nutrient availability
and limitation in Lake Ontario during IFYGL, Part II. Nitrogen available
in Lake Ontario tributary water samples and urban runoff from Madison,
Wisconsin. U. S. Environmental Protection Agency, Duluth, Minn., Eco-
logical Research Series. EPA-600/3-77-045. 88 p.
rivers, nutrients, nitrogen, algal assay, bottle test, algae, Selenastrum
capricornutum, Anabaena flos-aquae, Microcystis aeruginosa.
Samples of water from the Niagara, Genesee, Oswego, and Black Rivers were
collected from March to June, 1973. The samples were analyzed for nitrogen
forms and were incubated in darkness under aerobic conditions to promote
mineralization of soluble inorganic nitrogen from the organic nitrogen in the
samples. The amounts of ammonia and nitrate were determined as a function of
the time of incubation. Generally, over 50 percent of total nitrogen present
in these river samples was immediately available for algal growth or poten-
tially available after mineralization by bacteria. The results were highly
variable from each tributary and no single value could be selected from the
data obtained to describe the availability of total nitrogen in a given river.
(Abstract-Author).
0020-c
Cowen, W. F. , K. Sirisinha, and G. F. Lee. 1978. Nitrogen and phosphorus in
Lake Ontario tributary waters. Water, Air, and Soil Pollut. 10(3):343-
350.
rivers, streams, algal assay, phosphorus, nitrogen, nutrients, lake waters,
algae, Selenastrum capricornutum.
During the spring of 1972 through the spring of 1973 samples of rivers
tributary to Lake Ontario and streams in the Genesee River basin (New York)
were analyzed for N and P forms by chemical methods, then incubated in dark-
ness or bioassayed with algae to estimate the percentage of total N, organic
N, total P or particulate P which could eventually become available for algal
growth in Lake Ontario. The total available P in the river water samples
could be estimated by adding to the soluble orthophosphate 0.2 of the differ-
ence between the soluble orthophosphate and the total phosphate. The total
available N can be estimated from the sum of the inorganic N (NH3 and N03)
21
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plus 0.5 times the total organic N concentration of the river water sample.
(Abstract-Author).
0021
Czygan, F. C. 1970. Studies on the importance of the biosynthesis of secon-
dary carotenoids as a taxonomic character in green algae. Arch. Micro-
biol. 74(1):77-81.
biosynthesis, carotenoids, algae, Selenastrum gracile (strain 278-2), Chlor-
ella fusca (strain 211-8b).
Under certain extreme conditions, i.e. desiccation and UV-irradiation,
cells of the green chlorococcalean alga Selenastrum gracile (strain 278-2),
which normally produce secondary carotenoids (esters of astaxanthin and can-
thaxanthin) under nitrogen-deficient conditions, will be changed to forms
which are not able to biosynthesize such polyenes. The loss of this taxonomic
character will recover after 10-12 transfers in fresh culture medium. In
contrast, forms of Chlorella fusca (strain 211-8b) obtained as above do not
revert to the wild type producing secondary carotenoids, not even after 20
transfers. (Summary-Author).
0022
Denison, J. R. 1974. Limiting nutrient tests used in an investigation of
factors controlling phytoplankton development. Water Treat. Exam. 23(1):
52-75.
limiting nutrients, reservoirs, algal assay, ecological factors, phytoplank-
ton, algae, Selenastrum capricornutum.
Data are presented showing seasonal variations of phytoplankton in four
Manchester reservoirs and relationships are sought with some ecological fac-
tors controlling their growth. Selenastrum capricornutum was used as a test
species in bioassays to determine the availability of growth-1imiting algal
nutrients and to quantify the biological response to changes in concentrations
of these nutrients. Further study is required before precise interpretation
of the bioassay results can be provided, in relation to the naturally occurr-
ing algal populations, but the technique promises to be a valuable and versa-
tile tool for studying specific algal problems encountered in waterworks
practice. (Abstract-Author).
22
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0022-a
Dye, C., J. Hand, D. Jones, and L. Ross. 1978. Determination of critical
nutrient levels in Florida lakes using algal assays. Florida Dept. Envr.
Reg., Preliminary Report.
algal assay, lake waters, nutrients, eutrophication, agricultural runoff,
chlorophyll a, nitrogen, phosphorus, limiting nutrients, N:P ratios, Sele-
nastrum capricornutum. bottle test, algae.
The preliminary report is of a study conducted on 43 lakes in Florida.
Samples collected from the lakes were analyzed for chlorophyll a content,
total nitrogen, total phosphorus, and other chemical parameters. A bottle
test was conducted, using the alga Selenastrum capricornutum. The results are
given as critical chlorophyll a versus dry weight; and, limiting nutrients
(phosphorus was found to be the primary limiting nutrient). Twenty-seven of
the 43 lakes were found to have factors other than nitrogen and phosphorus
levels, that limited algal growth. (Abstract-Leischman-EPA).
0023
Ferris, J. J. , S. Kobayashi, and N. L. Clesceri. 1974. Growth of Selenastrum
capricornutum in natural waters augmented with detergent products in
wastewaters. Water Res. 8(12): 1013-1020.
phosphate, detergents, wastewater, lake waters, bottle test, algae, Sele-
nastrum capricornutum Printz, algal assay.
A determination of whether the removal of phosphate builders from deter-
gents would modify the ability of domestic secondary treated sewage effluent
to stimulate the growth of a test alga (Selenastrum capricornutum Printz) in
receiving waters alone and augmented with detergent products was made. The
lakes used as sources of test waters were located in northeastern New York
state and possessed total phosphorus concentrations ranging from ca. 0.001 to
0.04 mg P I-1.
The alga experienced stimulation in all three test lake waters from
secondary sewage containing detergent with phosphate or detergent without
phosphate. A concentration of 60 ug P I-1 was sufficient to effect signifi-
cant algal growth in two of the test waters; however, concentrations ranging
up to 110 ug P I-1 did not generate such a response in the third test water.
This latter result and others suggested that neither phosphorus nor other
23
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nutrients from these wastewater additions were the factors fully accountable
for the observed response(s). (Abstract-Author).
0024
Filip, D. S. , and R. I. Lynn. 1972. Mercury accumulation by the alga Sele-
nastrum capricornutum. Chemosphere 6:251-254.
mercury, algae, Selenastrum capricornutum, toxicity, algal assay.
Laboratory cultures of Selenastrum capricornutum were used to examine the
uptake of inorganic mercury. The test organism was exposed to amounts of
mercuric chloride to yield 0.1 ppm of mercury. The innoculated samples were
exposed to the mercury levels for two hours and then analyzed for mercury
content. The authors concluded that the mercury uptake of live cells (light-
incubated cultures) and formalin killed cultures were not statistically dif-
ferent. The uptake of mercury was concluded to be a passive absorptive pro-
cess. (Abstract-Leischman-EPA).
0025
Filip, D. S. , and E. J. Middlebrooks. 1975. Evaluation of sample preparation
techniques for algal bioassays. Water Res. 9(5/6):581-585.
pond water, membrane filtration, mesotrophic, eutrophication, algal assay,
pretreatment, feedlot runoff, algae, Selenastrum capricornutum, bottle test.
The chemical and biostimulatory properties of mesotrophic pond water and
eutrophic feedlot runoff were examined after subsamples of each were sepa-
rately subjected to standard membrane filtration and autoclaving pretreatments
for bottle test bioassays. Correspondingly separate subsamples were exposed
to ultraviolet light to determine its effectiveness as a possible pretreat-
ment. All three pretreatments fulfilled the requirement of destroying or
removing indigenous algae, but each technique substantially altered the water
chemistry changing the bioassay results. Filtration removed phosphorus;
autoclaving raised the pH value, precipated salts and eliminated dissolved
carbon dioxide; and ultra-violet light oxidized organic and inorganic nitrogen
compounds. (Abstract-Author).
24
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0026
Fitzgerald, G. P. 1970. Aerobic lake muds from the removal of phosphorus
from lake waters. Limnol. Oceanogr. 15(4):550-555.
phosphate, phosphorus, sorption rates, sediments, aerobic, phytoplankton,
Cladophora sp., algae, Selenastrum capricornutum. lake waters, limiting nutri-
ents, algal assay.
Phosphorus-limited Selenastrum and Cladophora sp. will respond by growth
or changes in extractable P04-P to as little as 0.02 mg P04-P in solution;
these same species did not respond when exposed for a period of 1 or 2 weeks
to as much as 2 mg of phosphorus as lake muds under aerobic conditions.
Studies of the rate of sorption of phosphorus by lake muds, under aerobic
conditions, from two lakes and from three depths in one lake, indicated that
as little as 0.4 g (dry wt.) of mud could sorb about 0.05 mg P04-P in less
than 30 min. These findings suggest that the sorption of phosphorus by lake
muds under aerobic conditions can be used to remove phosphorus from lake
water. (Abstract-Author).
0027
Fitzgerald, G. P. 1972. Bioassay analysis of nutrient availability, p.
147-165. Ijn H. E. Allen and J. R. Kramer [eds.], Nutrients in natural
waters, John Wiley & Sons, Inc. , New York.
algal assay, nutrients, phosphorus, nitrogen, algae, Microcystis aeruginosa,
Chlorella pyrenoidosa, Selenastrum capricornutum, Anabaena flos-aquae, am-
monia, lake waters, Spirogyra sp. , Cladophora sp.
The paper is a full-length chapter within the book, Nutrients i_n natural
waters, and consists of an in-depth discussion of the nutrient analyses for
bioassays. No abstract is provided. (Abstract-Leischman-EPA).
0027-a
Fitzgerald, G. P. 1974. Applications of growth and sorption algal assays.
U.S. Environmental Protection Agency, Washington, D. C. , Ecological
Research Series EPA-660/3-73-023. 175 p.
lake waters, nutrients, sorption, algal assay, phosphorus, agricultural run-
off, wastewater, water quality, algae, Selenastrum, Microcystis, Anabaena,
duckweed, Lemna minor, phytoplankton.
25
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The availability of nutrients in selected Wisconsin lakes was measured in
laboratory studies utilizing both sorption and growth algal assays. These
tests were conducted to evaluate contributions of phosphorus to the Madison
area lakes from septic tanks, agricultural runoff, and urban drainage and to
measure changes in the nutritional status of six lakes which were manipulated
for water quality improvement by nutrient inactivation or hypolimnetic aera-
tion. Characteristics of the assay techniques are discussed and results are
compared to chemical determinations of plant nutrients. (Abstract-Author).
0027-b
Fitzgerald, G. P. 1975. Are chemicals used in algae control biodegradable?
Water and Sewage Works 122(5):82-85.
pollution, biodegradation, toxicity, algistatic, algicidal, algal assay,
copper, mercury, ammonia, silver, algae, Selenastrum capricprnutuin, Micro-
cystis aeruginosa, Chlorella pyrenoidosa, Anabaena, Gloeotrichia, Oscilla-
toria.
Data collected during tests performed over the last 11 years have indi-
cated that the algicides that have been used to control the growth of problem-
causing algae, products containing copper, silver, mercuric chloride, phenyl
mercuric acetate, and quaternary ammonium compounds, do not pose a threat to
aquatic environments if used at the minimal concentrations required to be
effective, because these concentrations are biodegraded by the treated algae.
However, all chemicals to be added to aquatic environments should be evalu-
ated, since at least one compound --methyl mercuric chloride—is not detoxi-
fied by the action of treated algae and its use as an algicide would pose a
threat to the general environment. (Abstract-Author).
0027-c
Fitzgerald, G. P. 1975. Factors affecting the Algal Assay Procedure. Report
for Office of Research and Monitoring, U. S. Environmental Protection
Agency, Washington, D. C. Project No. P5J11912-J. 31 p.
algal assay, nutrients, bottle test, nitrogen, phosphorus, carbon, light
intensity, limiting nutrients, growth rates, standing crop, maximum yield,
fluorescence, lake waters, algae, Selenastrum capricornutum. phytoplankton,
Cladophora, diatoms.
26
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Evaluations of the Algal Assay Procedure (AAP) have demonstrated its
value for determining the level of nutrients in water samples which is avail-
able for the growth of algae as contrasted to chemical analyses of total
nutrient contents. The maximum specific growth rate, |j max, has been shown
not to be affected by the supply of N, P, or carbon when tests are carried out
using the suggested AAP. The maximum yield of algae is affected by the ini-
tial concentration of N and P (and Fe, Ca, Mg, K, and S in other nutrition
studies). The maximum yield is not affected by the supply of carbon in normal
AAP cultures (foam or cotton plugs), but increased incubation times might be
required for the maximum yield to be attained if comparatively concentrated
nutrient solutions are used. The inoculum levels of algae suggested in the
AAP are sufficiently low to make use of fluorometry as a measurement of the
growth of algae in relatively dilute culture media. If cell counts or absorb-
ance measurements are to be used to follow the growth of cultures, higher
initial cell densities may be employed. The p max of algae cultures is_ influ-
enced by the light intensity, but maximum yields merely require longer incuba-
tion periods if less than suggested light intensities are used. The applica-
tions of the AAP with the suggested algal species or HI vivo algae have demon-
strated its value in determining which algal nutrient will limit the growth of
algae in water samples. (Abstract-Author).
0027-d
Fitzgerald, G. P., S. L. Faust, and C. R. Nadler. 1973. Correlations to
evaluate the effects of wastewater phosphorus on receiving waters. Water
and Sewage Works. 120(1): 48-55.
phytoplankton, Rhizoclonium sp. , Cladophora sp. , Spirogyra sp. , algae, Sele-
nastrum capricornutum, phosphorus, rivers, wastewater, algal assay, bottle
test, nitrogen, iron, lake waters, sorption, extraction, chemical analysis.
Studies were conducted to determine the amounts of phosphorus that were
available for algal growth in lakes and rivers within Wisconsin. An algal
assay and a chemical analysis were the methods used to measure phosphorus
levels. Within the algal assay procedure, two tests were conducted, the
one-day sorption-extraction test and the long-term (12 days more) bottle test.
The results from the two tests were in agreement, as relating algal growth to
phosphorus availability. The authors did conclude that the long-term assay be
27
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used in conjunction with the short-term and the chemical analysis, as a check
system of the phosphorus availability for growth. The methods of measuring
available phosphorus were comparable; the authors concluded that either method
could be used to measure phosphorus concentrations available for algal growth.
The authors also concluded that data from the tests and analyses could be used
to determine whether water systems contained excess phosphorus. (Abstract-
Lei schman-EPA).
0027-e
Fitzgerald, G. P., M. S. Torrey, and G. C. Gerloff. 1975. Green Bay self-
purifies algal nutrients. Water and Sewage Works. 122(6):58-61.
pollution, wastewater, nutrients, phosphorus, nitrogen, water quality, algal
assay, bottle test, algae, Selenastrum capricornutum, phytoplankton, Clado-
phora, rivers, lake waters.
Algal assays were used to determine the effects of nutrient loadings in
the Fox River-Green Bay system. The studies were designed to aid in evalu-
ating and determining the effects of pollutants already present or being added
to the water system. Field assays were conducted with the green alga Clado-
phora; Selenastrum capricornutum, a green alga, was used in bottle tests to
determine effects of removal of phosphorus from the waters. The authors
concluded that if Green Bay were to be improved in terms of nutrient removal
(self-purification), the results would be effective in reducing algal growth
in the Great Lakes area. (Abstract-Leischman-EPA).
0027-f
Forsberg, A. 1974. Algal assay with drainage water from waste disposal.
Vatten 30(1):79-83. In Swedish.
pollution, wastewater, phosphorus, inhibition, algae, Ankistrodesmus falcatus,
Chlorella homosphaera, Scenedesmus quadricauda, Selenastrum capricornutum,
Euglena gracili.s, algal assay.
During spring 1972 some unknown pollution in the water of the River
Motala Strom gave the drinking water of Norrkoping a bad taste. Drainage
water from a big waste dump was discussed as a possible source for this inci-
dent.
28
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In order to study the effect of drainage water on algal growth, samples
were taken during a one year period, one week (5 days) every month, from a
brooklet receiving this drainage water. Mixed week samples were also analysed
for P04-P, total-P, COD and pH.
The algae were exposed to 0, 2.5, 10, 25 and 40% of the test water. The
incubation period was 7 days. The results are expressed as relative growth in
relation to the standard, an artificial nutrient solution without addition of
test water. Samples from the brooklet showed algal growth in proportion to
the amount of test water added. No toxic effects could be detected. The cor-
responding tests with water from the drainage pond gave inhibitory effects at
25% and 40%. (Modified translated Summary-Author).
0028
Forsberg, A. 1978. Research on recovery of polluted lakes: Chemical and
biological methods for monitoring wastewater effluent quality. Abstr.
Uppsala Diss. , ACTA Univ. Upsaliensis No. 458, Uppsala, Sweden. 28 p.
wastewater, toxicity, phosphate, phosphorus, ammonia, nitrite, nitrate, algae,
Ankistrodesmus, Chlorella, Scenedesmus, Selenastrum, Euglena, Minitest, algal
assay.
The quality of effluent water from chemical treatment plants has been
shown to vary considerably. Thus it is essential to conduct frequent sampling
to obtain a reliable basis for estimating nutrient loadings. The Minitest, a
handy method for sampling and analyzing phosphate, phosphorus, total phos-
phorus, chemical oxygen demand, ammonia, nitrite, nitrate, organic nitrogen
and other constituents if required, is presented. This procedure also in-
volves the use of a mixture of five algal species, four green algae and one
flagellate, for biological control of the effluent quality. Toxic effects of
wastewater effluent have been shown with a 40 times dilution (2.5% waste-
water). Besides description of fertilizing capacity and presence of toxic
compounds the algal assay also provides an opportunity to estimate the avail-
ability of nutrients, which for instance is of interest in comparisons between
different treatment processes. (Abstract-Author).
0028-a
Forsberg, A. 1978. Algal growth response and variation in cell composition
in cultures of a mixed algal system under varying nutrient conditions.
29
-------
No. V, Publ. in Abstr. Uppsala Diss., ACTA Univ. Upsaliensis No. 458,
Uppsala, Sweden. 14 p.
algal assay, nitrogen, carbon, phosphorus, chlorophyll a, algae, Ankistrodes-
mus falcatus (Corda) Ralfs., Chlorella sp., Scenedesmus quadricauda Turpin em.
Chod., Selenastrum capricornutum Printz, Euglena gracilis Klebs., nutrients.
A mixed algal system consisting of five algal species (Ankistrodesmus
falcatus (Corda) Ralfs, Chlorella sp. , Scenedesmus quadricauda Turpin em.
Chod., Selenastrum capricornutum Printz and Euglena gracilis Klebs) could
utilize at least 20 out of 31 organic nitrogen compounds as the only nitrogen
source. Analyses of cell contents of carbon, nitrogen, phosphorus, and chlor-
ophyll a gave the ranges 38-64%, 2.25-4.05%, 0.25-1.6% and 0.19-1.7% of dry
weight respectively. From the lower figures of nitrogen and phosphorus in
these ranges, cell yield coefficients could be estimated to be YN = 44 and Yp
= 400. The corresponding values obtained with gradient experiments gave about
twice the yields, YN = 90 and Yp - 775. (Abstract-Author).
0029
Forsberg, C. G. 1972. Algal assay procedure. J. Water Pollut. Control Fed.
44(8):1623-1628.
wastewater, nutrients, AGP, algae, Selenastrum capricornutum, Euglena
gracilis, Ankistrodesmus falcatus, Scenedesmus quadricauda, bottle test, algal
assay.
A new algal assay procedure using small samples, small culture volumes,
and short incubation periods has been developed. The algal growth potential
of wastewater effluents has been tested in a standard recipient solution
including test algae. The method permits tests with several algal species in
the same standard. The difference between the maximum algal growth in the
standard recipient solution with and without the addition of wastewater efflu-
ent was taken as a measure of the algal growth potential of the effluent.
Preliminary results obtained with effluents sampled at some Swedish wastewater
treatment plants are presented. The growth of Selenastrum capricornutum can
be about 90 percent lower in chemically treated wastewater than in effluent
from biological treatment. (Abstract-JWPCF).
30
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0029-a
Forsberg, C., and A. Claesson. 1974. Recovery of polluted lakes. Algal
growth limiting nutrients and algal assay (Selenastrum) chlorophyll.
Vatten 30(1):84-95. In Swedish.
wastewater, water quality, AGP, chlorophyll, N:P ratios, phosphorus, nitrogen,
chelation, trace elements, algal assay, algae, Selenastrum, limiting nutri-
ents.
This paper deals with algal assays performed parallel to the chemical
analyses with water collected from August—October, 1972. All algal assays
were performed with glass fiber filtered (Whatman GF/c, 0.45 u) surface water,
0—2 m. The algal assay procedure is essentially the same as the bottle test
method, though several modifications have been performed. Some are in accor-
dance with ideas previously presented e.g. small culture volumes. Also the
procedure for the enrichment experiments has been simplified. The tubes were
prepared beforehand with solutions of phosphorus, nitrogen, etc. in small
volumes. After evaporation the salt was resolved in the test water. A short
preliminary report of "Mini-test" with lake water has been presented.
The growth of the test alga, Selenastrum capricornutum, was determined as
the volume of the cells. The algal volumes have also been expressed as chlor-
ophyll. The correlation between the volume of Selenastrum and the chlorophyll
content (obtained by the acetone method) of the cells gave the correlation
coefficient r= +0.99 (n=25).
Enrichment experiments were performed in order to study algal growth
limiting nutrients. Results from Lake Kyrkviken show the variation of the
growth potential of surface water from week to week. Each sample tested was a
mixed week sample based on three samples/week. Enrichment with nitrogen had a
marked effect. Monthly average values of the growth potential of water from
Lake Si lien are given along with the results of tests enriched with phosphor-
us, nitrogen, chelating agent and trace elements.
The proportions between the lake water chlorophyll and the algal assay
(Selenastrum) chlorophyll are illustrated. The primary growth limiting nutri-
ents are also indicated. The sum of the natural lake water chlorophyll and
the algal assay chlorophyll has been related to the ratio total-N:total-P.
Lake Frosjon and Lake Glaningen showed special features. The former are
31
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polluted by a detergent factory which can explain the diverging conditions in
this lake water. The low chlorophyll content of Lake Glaningen can depend on
special conditions giving rise to big masses of very small easily consumed
algae. In any case the algal assay indicated no toxic effects in this water.
The sum of the two chlorophyll fractions mentioned above was correlated
to the content of total-P. Below phosphorus values of 0.05 mg/1 phosphorus
was the growth limiting nutrient. Above 0.1 mg/1 nitrogen played the corres-
ponding role. Between these values phosphorus, nitrogen or chelating agent
could be the primary growth limiting factor. The lakes belonging to the last
mentioned group, with total-P values between 0.05--0.1 mg/1 will be expected
to give the most rapid answer on advanced waste water treatment for phosphorus
removal. (Modified translated Summary-Author).
0030
Forsberg, C. G., and A. Forsberg. 1972, Algal growth potential test improves
sewage effluent control. Ambio 1(1):26-29.
wastewater, algal assay, phosphorus, algae, phytoplankton, Selenastrum capri-
cornutum, Euglena gracilis, Ankistrodesmus falcatus, Oocystis submarina var,
variabilis, Scenedesmus quadricauda, AGP, toxicity.
A new mixed algal assay procedure for monitoring sewage effluent is
described. Results from routine tests of effluents at treatment plants are
given. Five algae were used. Effluent from one plant showed daily rhythm in
algal production, low values during the summer vacation, and disturbed treat-
ment efficiency due to incoming oil. The correlation coefficient for total-P
and algal volume was + 0.81; for P04-P and algal volume it was + 0.99. Toxic
effects were noted. (Abstract-Ambio).
0030-a
Forsberg, C. , and E. Hokervall. 1971. An algal assay procedure of value for
control of sewage effluents from treatment plants in Stockholm. Vatten
27(l):51-57. In Swedish.
wastewater, algal assay, AGP, algae, Selenastrum capricornutum, chemical
flocculation.
This paper presents the first results with a new algal assay procedure of
value for control of sewage effluents from treatment plants with chemical
32
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flocculation. In July, 1970, about 30 chemical treatment plants were func-
tioning in Sweden and more than 200 more were projected. Since the aim of
chemical flocculation is to reduce the fertilizing power of the effluents, the
idea of using an algal-assay procedure has been tested at the algal test
laboratory of the Institute of Physiological Botany, Uppsala. A brief des-
cription of the method is presented together with some results obtained with
effluents from three treatment plants in Stockholm.
Small samples of the effluents were frozen quickly and then sent by
regular mail while still frozen, in special boxes. After thawing and filter-
ing, the samples were diluted in a recipient solution with ionic concentra-
tions of the same order as ordinary lakes. The recipient solution also in-
cludes the test alga, Selenastrum capricornutum (a mixed algal standard will
be developed later). To this standard the sewage effluent was added at con-
centrations of 5 o/o, 2.5 o/o or lower. The difference between the maximal
algal growth in the standard recipient with and without addition of sewage
effluents were calculated. The volume of algae produced was measured by
electronic particle counting. The new method requires small volumes of sam-
ple, about 5 ml, small culture volumes, 1 ml, and short test periods, 3--4
days.
Algal-growth potential (AGP) values from some tests with sewage effluents
from the chemical-biological treatment plant at Eolshall are presented. The
AGP values were transformed from the algal volume figures. AGP-values between
0.5--1.5 were obtained when no disturbance of the treatment was detected.
During the last part of April the spring flood was the reason for the poor
treatment which gave increased AGP-values. At the end of September and begin-
ning of October the values reached a level of about 8 which corresponds to
biological treatment only. This depended on incoming oil (September 28--29)
and technical errors (first half of October) with the equipment for the chemi-
cal flocculation. The biological treatment plants of Loudden and Akeshov-
Nockeby were tested during the end of their operation as biological treatment
plants only. The plant at Akeshov-Nockeby gave values of the same order as
other biological treatment plants tested, about 8, while the plant at Loudden
reached higher values. On December 1, 1970 the plant at Akeshov-Nockeby
started with chemical-biological treatment and for the last period tested,
December 3--9, all AGP values were between 0.5--2. (Modified translated
Summary-Author).
33
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0030-b
Forsberg, C. , and E. Hokervall. 1972. Algal growth potential test (AGP-test)
of sewage effluent. Vatten. 28(l):17-26. In Swedish.
wastewater, AGP, algal assay, nutrients, nitrogen, phosphorus, phosphate,
algae, Ankistrodesmus falcatus, Euglena gracilis, Oocystis submarina var.
variabilij, Scenedesmus quadricauda, Selenastrum capricornutum.
From February 1971 the efficiency of sewage treatment at the chemical and
biological treatment plant at Akeshov-Nockeby (230,000 pe.), Stockholm, has
been continuously tested using a new algal assay procedure. Results from 240
day-samples together with chemical data are presented.
The nutrient content of unfiltered sewage effluent was tested after
dilution to 2.5 o/o with a laboratory recipient solution. This solution
includes the test algae, five different species: Ankistodesmus falcatus,
Euglena gracilis, Oocystis submarina var. variabilis, Scenedesmus quadricauda
and Selenastrum capricornutum. These species were chosen for their excellent
growth in dilute sewage. The difference between maximal algal growth in the
standard with and without the addition of sewage effluent was taken as a
measure of the algal growth potential (AGP). The method for the algal assay
procedure, which will subsequently be described in more detail, requires
comparatively short test periods, small volumes of sample (5 ml), small cul-
ture volumes (2.5 ml) and employs disposable plastic culture tubes.
Chemical analyses of the day-samples representing Tuesday to Wednesday
were performed, parallel to the algal tests at The Stockholm City Municipal
Services Department, Construction Division. Phosphate-phosphorus was analysed
at the algal test laboratory as day-average per week for 7 mixed day-samples.
The algal volumes obtained during 8 months are given. During these
months the treatment plant showed a rhythm. The high values in June are due
to incoming oil, which disturbed the last (biological) step in the treatment.
During July and August some samples gave no algal production at all indicating
some inhibitory effect of the effluent. The phosphate content was extremely
low during the vacation period, with some average values per week below 0.10
mg/1. In nonindustrial effluent from other treatment plants tested, the same
results were obtained when the Al content was comparatively high, 1--2 mg/1,
with the highest value of 4 mg/1. As preliminary studies have indicated that
algal growth can be retarded by comparatively low amounts of Al (unpublished),
34
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one reason for the low production in the effluent, as observed during July and
August, could be that the dosage of Al-sulphate in the treatment plant was too
high in relation to the existing, low phosphate content. Correlations for
monthly average values for samples representing Tuesday—Wednesday were ob-
served between total-phosphorus, phosphate-phosphorus and algal volume pro-
duced. The correlation coefficients were +0.81 and +0.88, respectively.
Positive correlations also were obtained between the monthly average values of
BOD5, suspended material and total-phosphorus with the correlation coeffi-
cients of +0.78 and +0.89 respectively. The relations between total-nitrogen,
total-phosphorus and algal volume produced are illustrated. The correlation
between algal volume, total-phosphorus and BOD5, is also outlined.
The average algal volume produced during the whole period was 425 mm3/!
(n=240). The corresponding phosphate value was 0.25 mg/1. The biggest out-
flow of phosphate-phosphorus occurred during week 24, when 500 kg was trans-
ported by 760,000 m3 effluent. During week 31 only 60 kg was transported by
700,000 m3.
As a result of this investigation further effort will be directed into
decreasing the fertilizing capacity of the effluent and to eliminating the
high values obtained in the middle of the week. During the vacation period in
1972 the dosage of Al-sulphate will be lowered from that of the normal months.
(modified translated Summary-Author).
0031
Forsberg, C. G. , S.-O. Ryding, and A. Claesson. 1975. Recovery of polluted
lakes: A Swedish research program on the effects of advanced wastewater
treatment and sewage diversion. Water Res. 9(1):51~59.
Minitest, wastewater, algae, Selenastrum capricornutum, chlorophyll a, phos-
phorus, nitrogen, lake waters, algal assay.
In order to elucidate the effects of the effluents from advanced waste-
water treatment plants for phosphorus removal, the National Swedish Environ-
ment Protection Board during 1972 started a program for analyzing the condi-
tions in a number of different recipient lakes. This study of recovery of
polluted lakes also includes some recipient lakes from which sewage effluent
will be completely diverted. This paper presents the program, some new meth-
ods used and the treatment plants and recipient lakes involved. The project,
35
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including 18 lakes and 15 sewage treatment plants is running in cooperation
with 13 Provincial Governments and 16 Municipalities. The efficiency of the
sewage treatment plants is continuously monitored by a new method called
Minitest, and the nutrient loadings from these plants as well as from other
sources are calculated.
As the water quality of many recipient lakes showed great variations
during the vegetation period, a need for a frequent sampling exists in these
lakes.
In order to simplify the sampling procedure sampling on the shore (after
pumping water from the sampling point) has been introduced. Data obtained
after analyzing samples taken from a boat and samples from the same area taken
on the shore after pumping through a 600 m plastic tube showed good agreement.
The lake water samples are preserved by freezing. Analyses after rapid
thawing showed that this method can be used also when determining from cell
material easily leaking ions such as P04, N03, etc.
The sewage treatment plants, the recipient lakes and some background data
are presented. The water quality of the lakes is illustrated with the corre-
lation between chlorophyll a and suspended solids. The correlation coeffi-
cient = + 0.88. The first sign of lake recovery has been observed after an 85
percent reduction of the waste phosphorus load. Post-precipitation (Al-
sulphate) decreased the yearly load from 2000 kg P to 300 kg, which gave a 50
percent reduction of the P content of the recipient water. Also, the chloro-
phyll values decreased.
The role of nitrogen and phosphorus as algal growth limiting nutrients
has been studied by using an algal assay procedure. N limited the growth of
Selenastrum capricornutum in 2/3 of the samples. In filtered lake water P was
the growth limiting nutrient at total-P values below 0.05 mg I-1. Above 0.1
mg P I-1, N played the principal role. Between these values the growth was
primarily limited by P or N or chelating agents. (Abstract-Author).
0031-a
Forsberg, C. , S.-O. Ryding, A. Forsberg, and A. Claesson. 1978. Research on
recovery of polluted lakes. I. Improved water quality in Lake Boren and
Lake Ekoln after nutrient reduction. Verh. Int. Ver. Limnol. 20(2):825-
832.
36
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wastewater, phosphorus, nitrogen, chlorophyll a, COD, algal assay, algae,
Selenastrum capricornutum, water quality, nutrients.
After advanced wastewater treatment for P removal improved water quality
has been observed in Lake Boren and Lake Ekoln. The P-load from the treatment
plants at Motala and Uppsala has been reduced from 30 to 3 and from 100 to 20
tons yr-1, respectively. In L. Boren and L. Ekoln the average chlorophyll a
values for surface water have decreased by 50 o/o, from about 10 to 5 and from
20 to 10 mg m-3, respectively. For lowering chlorophyll a concentration by 1
mg m-3 in the lakewater as an average for the summer period, the annual P-load
had to be decreased by 6 and 5 tons, respectively. The P residence time was
of the same order in these lakes. In both lakes the transparency as an aver-
age for the summer period has increased from 1.5 to 2 m. Further improvements
can be expected. (Summary-Author).
0032
Forsberg, C. , S.-O. Ryding, A. Claesson, and A. Forsberg. 1978. Water chemi-
cal analyses and/or algal assays? Sewage effluent and polluted lake
water studies. Mitt. Int. Ver. Limnol. 21:352-363.
wastewater, algal assay, lake waters, chemical analysis, limiting nutrients,
algae, Selenastrum capricornutum, Ankistrodemus falcatus, Scenedesmus quad-
ricauda, Chlorella sp. , Euglena gracilis, AGP, pollution.
In a Swedish research program dealing with recovery processes of polluted
lakes, algal assay procedures have been routinely used for three years, for
determining algal growth potential (AGP) and growth-1imi ting nutrients. Both
sewage effluent and lake water were studied. Algal assay data and results
from chemical water analyses are presented in order to illustrate and discuss
the usefulness of the practical application of algal assay procedures.
After chemical treatment for phosphorus removal, the algal growth of five
species (mixed culture of Ankistrodesmus falcatus, Chlorella homosphaera,
Euglena graci1 is, Scenedesmus quadricauda, and Selenastrum capricornutum) in
sewage effluent, often showed a high correlation to the phosphate content of
the test water. But, the results also indicated both growth-inhibiting and
toxic compounds in the effluent. Monthly average values for nitrogen, phos-
phorus, chlorophyll a (chl a) and AGP expressed a chl a, are presented from
three lakes. The AGP values of filtered water, expressed as chl a, were
37
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determined for 17 polluted lakes by using S. capricornutum as the test alga.
In four lakes, AGP was < 10 mg chl a/m3. The maximum value obtained was 488
mg/m3.
The Selenastrum test indicated N to be the most limiting nutrient in
seven of 17 lakes, P in seven lakes, and N, P in the remaining three. Calcu-
lated from chemical data, N was found to be the most limiting nutrient in five
lakes and P in eight lakes. In four lakes, the available P and N was calcu-
lated to give rise to the same AGP values, which means that in these lakes
neither P nor N could be looked upon as the most limiting nutrient. Ratios
for N:P in the lake water in relation to the role of these elements as most
limiting nutrient are suggested. Values obtained for the maximum specific
growth rate of Selenastrum showed tendencies to increasing growth rate with
increasing degree of pollution.
Algal assays were found to be of great value for explaining situations
which cannot be indicated by chemical analysis. On the other hand, many
results presented today by algal assays can equally well be calculated by
water chemical data. (Modified Summary-Author).
0033
Francisco, D. E. , and C. M. Weiss. 1973. Algal response to detergent phos-
phate levels. J. Water Pollut. Control Fed. 45(3):480-489.
detergents, phosphate, wastewater, algal assay, phosphorus, eutrophication,
lake waters, algae, Selenastrum capricornutum, limiting nutrients.
Algal assays were performed on treated wastewaters, some containing
phosphate detergents and others containing a non-phosphate detergent. In
general, phosphate detergent wastewater allowed the same growth as did non-
phosphate detergent wastewater at each level of treatment. Tertiary treatment
for phosphorus removal resulted in much less growth than secondary treatment.
Tertiary-treated phosphorus detergent wastewater allowed significantly greater
growth than the non-phosphorus wastewater at a concentration of 2 percent.
Simply removing detergent phosphates from wastewater apparently will not
control eutrophication, except where the receiving water is very strongly
phosphorus-limited and wastewater is the principal phosphorus source, but may
allow significantly better tertiary phosphorus removals. (Abstract-JWPCF).
38
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0034
Funk, W. H., F. W. Rabe, R. Filby, J. I. Parker, J. E. Winner, L. Bartlett, N.
L. Savage, P. F. X. Dunigan, Jr., N. Thompson, R. Condit, P. J. Bennett,
and K. Shaw. 1973. Biological impact of combined metallic and organic
pollution in the Coeur d'Alene - Spokane River drainage system. Washing-
ton State Univ. University of Idaho. OWRR (B-044 WASH and B-015 IDA).
187 p.
rivers, sediments, toxicity, zinc, algae, Selenastrum capricornutum, phyto-
plankton, algal assay, water quality, cadmium, lead, mercury, iron, manganese.
An investigation was made into the biological effects of metallic and
organic pollution in the Coeur d'Alene - Spokane River Lake drainage system.
Water quality data show heavy inflows of nutrients during late spring
into Coeur d'Alene Lake from the St. Joe and Coeur d'Alene Rivers and Plummer
Creek. Although no waters devoid of dissolved oxygen have been measured, 2-4
mg/1 have been measured in the southern portion of the lake. Phytoplankton
productivity measurements and bottom organism identification and enumeration
have shown that the lake functions at several trophic levels, meso-eutrophic
to eutrophic in the southern portion, mesotrophic in the immediate area of the
Coeur d'Alene River and oligotrophic in the northern portions. High metal
concentrations (1000-7000 mg/kg Zn) in the bottom sediments of the middle and
northern portions of the lake did not appear to substantially affect distri-
bution of benthic chironomids or oligochaetes.
Lake sediment cores taken across the delta region of the Coeur d'Alene
River penetrated through the regions of high metallic concentration and al-
lowed an average sediment deposition rate to be calculated.
Algal toxicity tests showed the amount of Zn normally present in the
Coeur d'Alene Lake and River and the Spokane River to be inhibitory to the
algal test organism Selenastrum capricornutum.
The water quality of the Spokane River was shown to be of good to excel-
lent quality in all parameters tested except for high metallic content, es-
pecially Zn. Activation and atomic absorption analysis of the metallic con-
tent in tissues of the organisms populating the Spokane River indicated that
the algae were the prime concentrators of Zn, Cd, Pb, Hg, Fe, and Mn. Algae
and detritus consumers such as the larvae of the caddis fly Hydropsyche and
the nymphs of may fly Baetis reflected high metallic concentrations. Most
higher aquatic plants showed relatively lower concentrations.
39
-------
Analysis of fish tissues showed a considerably less concentration of
metals than the aquatic-plants, insects, or algae. However, Zn was measured
at concentrations of 80-200 mg/kg in liver tissues of several species of
fishes. Fillet tissues generally contained less than one quarter of these
amounts. The fish, when collected, did not appear to be under stress leading
the investigators to believe that most of the metals, although present in
relatively high concentration in the tissues of the organisms tested, must be
in a relatively innocuous state. (Abstract-Author).
0035
Gargas, E. 1973. Preliminary statements of algal assay procedure at the
Water Quality Research Institute, Denmark, p. 19-32. Ln Algal assays in
water pollution research, Proc. Nordic Symp., 25-26 October 1972, Oslo,
Norway. NORDFORSK, Secretariat of Environmental Sciences.
algal assay, bottle test, algae, Selenastrum capricornutum, Phaeodactylum
tricornutum, AGP, diatoms.
A modified bottle test procedure was used to determine how algae would
grow under different cultivating conditions, such as light intensity. The
author found no difference in biomass (when expressed as cell number, cell
volume, organic nitrogen or organic phosphorus) cultivated under different
light intensities, although the cultures under the lowest light intensity
showed a growth lag. The author indicated that light conditions must be
considered when expressing biomass in terms of chlorophyll a content or inor-
ganic carbon. The algal species used were Selenastrum capricornutum and
Phaeodactylum tricornutum. (Abstract-Leischman-EPA).
0036
Gargas, E. 1978. The effect of sewage (mechanically, biologically, and
chemically treated) on algal growth. Mitt. Int. Ver. Limnol. 21:110-
124.
wastewater, algal assay, rivers, lake waters, nitrogen, phosphorus, algae,
Selenastrum capricornutum.
The report is a presentation and discussion of results of algal assays
conducted in a laboratory and j_n situ. The assays were part of an investi-
gation on pollution of the Guden River System in 1974-75. The laboratory
40
-------
experiments were designed to study effects on growth rates of Selenastrum
capricornutum from spikes of increasing concentrations of sewage (mechanically
treated and biologically-chemically treated). Organic matter produced was
measured and expressed in terms of cell numbers, dry weights, chlorophyll a
content, and concentration of nitrogen and phosphorus for each concentration
of sewage spike. The addition of the sewage spikes into the natural system
caused an increase in phytoplankton production but the author could not con-
clude that the production was directly a result of increased growth rates.
Another form of measurement used in the field experiments was the combination
of secchi depth and chlorophyll a content. The author was able to make linear
regression analyses to estimate the effect of sewage loadings on the increased
chlorophyll a content. The author concludes that the algal assay could be
conducted with sewage waters, as spiked and unspiked waters, to estimate the
effects of sewage loading on receiving waters; however, it was not possible to
estimate the effects of sewage loading on the natural phytoplankton in the
field. (Abstract-Leischman-EPA).
0037
Garton, R. B. 1973. Biological effects of cooling tower blowdown, p. 284-
292. In Water-1972, AIChE Symp. Ser. No. 129, Vol. 69, New York.
algae, Selenastrum capricornutum, algal assay, fish, Salmo gairdneri, morpho-
line, cyclohexylamine, hydrazine, zinc, phosphate, sulfates, sodium chromate,
heavy metals, toxicity: nuclear power.
Toxicity was determined of a 'typical' cooling tower blowdown and of its
individual components. Since blowdown composition is variable, a simulated
blowdown was made using chemicals and concentrations listed in waste discharge
permit applications. The mixture contained varying amounts of chromate, zinc,
phosphate, sulfate, boron, sodium, ammonia, morpholine, cyclohexylamine, and
hydrazine, and simulated blowdown from the Trojan nuclear plant. Separate
tests were also conducted with sodium chromate, zinc, phosphate, and with
blowdown without chromate. Fish (Salmo gairdneri) and an alga (Selenastrum
capricornutum) were used in bioassays of the mixtures. Algal assays were
conducted according to the Algal Assay Procedure Bottle Test. Fish bioassays
were conducted in accordance with procedures given in Standard Methods. Algal
growth was reduced by blowdown at full strength and at dilutions of 0.1. Fish
41
-------
were killed at dilutions of 0.1. No effect was observed at 0.032 dilution.
The LC50 value was 0.068 dilution. With sodium chromate, algal growth was
reduced at 0.139 ppm, and "fish survived 96 hours at 31 ppm. Zinc phosphate
had no significant effect on algal growth at 0.016 ppm. The LC50 for fish was
0.09 ppm zinc phosphate. Based on all the tests, zinc and chromium were
identified as the toxic components in the blowdown. It is noted that syner-
gistic effects may occur in other mixtures containing different compounds.
Furthermore, the organisms used in the bioassays mus't be specified since
toxicities may differ for different ones. (Abstract-Little-Battelle WRSIC).
0038
Gerhold, R. M. 1976. Algal nutritional bioassay of Lake Wylie, North Carol-
ina, p. 175-220. In E. J. Middlebrooks, D. H. Falkenborg, and T. E.
Maloney [eds.], Biostimulation and nutrient assessment, Ann Arbor Sci.,
Mich.
lake waters, algal assay, bottle test, algae, Selenastrum capricornutum,
limiting nutrients, phosphorus, phytoplankton, Anabaena circinalis, Anabaena
wisconsinense, Microcystis aeruginosa, Anabaena scheremetievi, Anabaena sprio-
ides, Raphidiopsis curvata, Oscillatoria tenuis, Oscillatoria geminata.
The key findings from the bioassay studies, ancillary laboratory experi-
ments, and the field observations are summarized below:
1. Phosphorus was identified as a limiting nutrient in Lake Wylie
waters.
2. Overall, it appears that an approximate doubling of the mean annual
available phosphorus concentration in the Lake Wylie study area would have to
occur before another nutrient would become limiting.
3. The bioassays did not indicate any influence of the Allen Stream
Station on the ability of the Lake Wylie study area waters to support algal
growth as a function of nutrient content.
4. No measurable effects of urbanization were encountered in the study.
5. Ancillary laboratory experiments and field observations support the
hypothesis that the Lake Wylie study area waters are not conducive to the
overabundant growth of potential nuisance blue-green algae, and that the low
hydrogen ion concentration may be prominent among the factors responsible.
6. Field observations suggest that the genus Oscillatoria was the most
prominently represented blue-green algae during the study period. (Summary-
Author).
42
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0039
Gerhold, R. M. , and R. G. Otta. 1976. Algal bioassays to evaluate a proposed
cooling lake. J. Water Pollut. Control Fed. 48(10):2351-2366.
lake waters, reservoirs, algal assay, algae, Selenastrum capricornutum, nitro-
gen, phosphorus, nuclear power.
The primary objectives of this study of waters to be used in supplying a
proposed cooling lake for a Kansas nuclear power station were to identify and
quantify algal growth-limiting nutrients, compare available concentrations
with chemical analyses, predict sensitivity to changes in concentrations of
the limiting nutrients, measure the algal growth potentially supported by the
waters, and predict the effects on algal growth of evaporative concentration
of nutrients within the proposed impoundment.
The results support the following conclusions:
1. Nitrogen or phosphorus alone or in association were the limiting
nutrients. Some differences in limiting nutrient relationships were noted in
samples collected at different times of the year.
2. Chemical analysis of the inorganic forms such as orthophosphate
phosphorus or the sum of ammonia, nitrate, and nitrite nitrogen frequently
underestimated the biologically available portion of the limiting nutrient,
and total phosphorus, or total nitrogen as the sum of organic and inorganic
nitrogen forms generally overestimated the biologically available portion.
For this reason, it was concluded that chemical analyses alone should not be
substituted as estimators of biological availability.
3. Although added nitrogen often may lead to increased algal growth in
the test waters, phosphorus should be considered the most economical key to
controlling future eutrophication in the proposed cooling lake or in the
source waters for the lake.
4. The data from experiments repeated during four quarters of an annual
cycle suggest that evaporation more than two-fold may lead to a significant
reduction in algal production, rather than an increase as might have been
predicted for evaporative concentration of nutrients.
5. It was concluded that the procedure, as applied in this study should
be useful in understanding the long-range effects, if any, of power plant
operation. The technique is simple. Equipment and the required laboratory
facilities are uncomplicated and economically modest. (Conclusions-Author).
43
-------
0040
Goldman, J. C. 1976. Effects of temperature on growth constants of Sele-
nastrum capricornutum. J. Water Pollut. Control Fed. 48(9): 2215-2216.
temperature, algae, Selenastrum capricornutum, growth rates, ammonia, algal
assay.
The report is a discussion of the basis behind discrepancies between two
studies of growth constants and growth rates as functions of temperature for
the alga Selenastrum capricornutum. One study was conducted by the author,
Goldman et a\_. and the other study was conducted by Reynolds et a/L (Ab-
stract- Lei schman-EPA).
0041
Goldman, J. C. , W. J. Oswald, and D. Jenkins. 1974. The kinetics of inor-
ganic carbon limited algal growth. J. Water Pollut. Control Fed. 46(3):
554-574.
carbon, inorganics, kinetics, pH, continuous flow, algae, Selenastrum capri-
cornutum, Scenedesmus quadricauda, algal assay.
A series of continuous culture studies under inorganic carbon limiting
conditions with axenic cultures of two representative green algae were per-
formed in the pH range 7.05 to 7.61. Kinetic coefficients were determined for
both algae. It was found that the inorganic carbon limited specific growth
rates of both species were described by the Monod model when bicarbonate
alkalinity was the sole inorganic carbon source. It was also found that
inorganic carbon limited algal growth is a function of the total inorganic
carbon present. Half-saturation coefficients for each alga were found to be
low in relation to the quantities of inorganic carbon available from bicar-
bonate alkalinity in most natural waters. It was found that these coeffi-
cients increased significantly, however, with small increases in pH. (Ab-
stract- JWPCF).
0042
Goldman, C. R., M. G. Tunzi, and R. Armstrong. 1969. Carbon-14 uptake as a
sensitive measure of the growth of algal cultures, p. 158-170. In E. J.
Middlebrooks, T. E. Maloney, C. F. Powers, and L. M. Kaack [edsTT Proc.
Eutroph. Bioassess. Workshop, 19-21 June 1969, U.S. Pacific Northwest
Water Laboratory, Corvallis, Oregon.
44
-------
carbon-14, algal assay, algae, Chlorella pyrenoidosa, Selenastrum gracile,
wastewater, limiting nutrients.
The report is a summarization of results from experiments designed to
test the reliability of the carbon-14 uptake method as a measurement of algal
production. The method was then compared to other methods of measuring algal
growth such as cell counts, absorbance as cells multiply, and determination of
biomass as cells are produced. The algal species used were Chlorella pyre-
noidosa and Selenastrum gracile. A series of experiments were conducted using
filtered effluent from Reno-Sparks Sanitary Treatment Plant and Upper Truckee
Trout Creek as additions to filtered Lake Tahoe water. Each method, cell
counts, optical density, and carbon-14 uptake were used as measurements of
algal growth. The author found that the Chlorella experiments provided a good
comparison of the methods. The growth of the Chlorella culture was measured
and expressed adequately by using the carbon-14 method. The author concluded
that the carbon-14 method was easy to use for measuring algal growth within
nutrient-rich and nutrient-poor waters. The carbon-14 method experiments
could detect growth in waters with natural phytoplankton when the other meth-
ods gave no indication of change in biomass. The author also found that the
carbon-14 method yielded values proportional to the mass of cells in the
cultures. (Abstract-Leischman-EPA).
0043
Greene, J. C. , W. E. Miller, T. Shiroyama, and T. E. Maloney. 1975. Utili-
zation of algal assays to assess the effects of municipal, industrial and
agricultural wastewater effluents upon phytoplankton production in the
Snake River System. Water Air Soil Pollut. 4:415-434.
algal assay, rivers, algae, Selenastrum capricornutum, wastewater, toxicity,
nitrogen, phosphorus, inhibition, bottle test.
The most characteristic water quality problem of the Snake River Basin is
the excessive aquatic growth and thick blooms of algae. The cause of these
aquatic growths is related to the high concentrations of basic nutrients -
nitrogen and phosphorus - in the Snake River system. Industrial wastes,
natural phosphate levels, irrigation return flows, municipal wastes, and the
decay of aquatic biota all contribute to the nutrient balance which stimulates
aquatic growths.
45
-------
Algal assays were conducted on waters of eighteen Snake River and tribu-
tary sites to: (1) determine if algal growth was consistent with results
predicted from review of chemical analysis for orthophosphorus and total
soluble inorganic N; (2) determine if algal yields were limited to P, N, or
some other nutrient essential to algal growth; and (3) predict the effects of
N or P additions in algal productivity. (Abstract-Author).
0044
Greene, J. C. , W. E. Miller, T. Shiroyama, and E. Merwin. 1975. Toxicity of
zinc to the green alga Selenastrum capricornutum as a function of phos-
phorus or ionic strength, p. 28-43. In Proc. Biostim. Nutr. Assessmt.
Workshop, 16-17 October 1973, U.S. Environmental Protection Agency,
Con/all is, Oregon. EPA-660/3-75-034.
algal assay, algae, Selenastrum capricornutum, toxicity, zinc, phosphorus,
inhibition, heavy metals, bottle test.
Experiments are described in which a modifiction of the algal assay
procedure bottle test was used to evaluate Selenastrum capricornutum as a
bioassay for zinc toxicity and to determine whether relationship exists be-
tween zinc toxicity and phosphorus concentrations that would affect the 14-day
maximum growth yield of this alga in a defined inorganic medium. Results
indicated that phosphorus (as orthophosphorus) concentrations of 0.047 to
0.930 mg/1 did not significantly affect the toxicity of zinc to this alga and
that algal cell numbers did not significantly affect the concentration of zinc
required to produce 95% inhibition of the 14-day maximum yield. Ionic
strength (specific conductance) was the dominant factor regulating zinc tox-
icity as the sensitivity of the test alga to zinc was inversely proportional
to the increase in ionic strength of the test substrates. The factor of 2.72
plus or minus 20% multiplied by the ionic strength (micromhos/cm) of a test
substrate will indicate the level of zinc (in microgram/1) that would inhibit
95% growth of the alga provided other antagonistic or synergistic constituents
are absent. (Abstract-Auen-Wisconsin WRSIC).
0044-a
Greene, J. C. , W. E. Miller, T. Shiroyama, R. A. Soltero, and K. Putnam.
1976. Use of algal assays to assess the effects of municipal and smelter
wastes upon phytoplankton production, p. 327-335. In Proc. Symp. Terr.
Aquat. Ecol. Studies of N.W., EWSC Press, Cheney, Wash.
46
-------
algal assay., bottle test, algae, heavy metals, lake waters, wastewater, riv-
ers, zinc, water quality, toxicity, Selenastrum capricornutum.
The Algal Assay Procedure Bottle Test is used as a tool for defining and
understanding water quality management problems including: (1) the effects of
municipal and industrial waste discharges upon algal productivity; (2) the
effects of removing inhibitory or toxic substances from wastewaters on algal
productivity when nutrients are not controlled; and (3) the degree to which
wastes must be controlled to enhance water quality. The algal assay procedure
is currently being used to determine the combined effects of smelter waste
high in zinc content and domestic waste inflow from the City of Spokane, Wash-
ington, upon algal growth in the Spokane River basin. Long Lake, an impound-
ment of the Spokane River, is highly eutrophic as a result of nutrient input
from the city's primary sewage treatment plant. Throughout the study Long
Lake was limited for optimal algal growth primarily by either nitrogen or
phosphorus. Continued presence of heavy metals in the system was indicated by
increased yields of Selenastrum capricornutum in test waters spiked with ethy-
lenediaminetetraacetic acid (EDTA). For approximately 80 years, heavy metal
inflow throughout the system has probably affected species composition but not
productivity in the natural environment. (Abstract-Author).
0045
Greene, J. C. , W. E. Miller, T. Shiroyama, R. A. Soltero, and K. Putnam.
1978. Use of laboratory cultures of Selenastrum, Anabaena and the indi-
genous isolate Sphaerocystis to predict effects of nutrient and zinc
interactions upon phytoplankton growth in Long Lake, Washington. Mitt.
Int. Ver. Limnol. 21:372-384.
water quality, algal assay, zinc, nitrogen, phosphorus, algae, Anabaena flos-
aquae, Selenastrum capricornutum, Sphaerocystis schroeteri, phytoplankton,
bottle test, wastewater, rivers, lake waters.
The Algal Assay Procedures Bottle Test is a tool to define and understand
water quality management problems including: (1) the effects of municipal and
industrial waste discharges upon algal productivity; (2) the effects of re-
moving inhibitory or toxic substances from wastewaters on algal productivity
when nutrients are not controlled; and (3) the degree to which wastes must be
controlled to enhance water quality. The algal assay procedure is currently
being used to determine the combined effects of smelter waste high in zinc
47
-------
content and domestic waste inflow from the City of Spokane, Washington, upon
algal growth in the Spokane River basin. Long Lake, an impoundment of the
Spokane River, is highly eutrophic as a result of nutrient input from the
city's primary sewage treatment plant. Throughout the study, Long Lake was
limited for optimal algal growth primarily by either phosphorus or nitrogen.
Continued presence of heavy metals in the system was indicated by increased
yields of Selenastrum and Anabaena in test waters spiked with ethylenedini-
trilotetracetic acid (EDTA); its addition was not required to support optimum
growth of Sphaerocystis. For approximately 80 years, heavy metal inflow
throughout the system has probably affected species composition but not pro-
ductivity in the natural environment. (Abstract-Author).
0046
Greene, J. C., R. A. Soltero, W. E. Miller, A. F. Gasperino, and T. Shiroyama.
1976. The relationship of laboratory algal assays to measurements of
indigenous phytoplankton in Long Lake, Washington, p. 93-126. In E. J.
Middlebrooks, D. H. Falkenborg, and T. E. Maloney [eds], Biostimulation
and nutrient assessment, Ann Arbor Sci. , Mich.
algal assay» lake waters, algae, Selenastrum capricornutum, indigenous phyto-
plankton, heavy metals, bottle test, zinc, phosphorus, nitrogen, limiting
nutrients, chlorophyll, reservoirs, eutrophication.
To determine if laboratory algal assays of natural waters actually re-
flect environmental conditions, studies were made of Selenastrum capricornutum
growth potential on autoclaved and filtered water from Long Lake, Washington.
Increased algal yields in EDTA spiked test waters showed toxic levels of heavy
metal(s). Maximum algal yields were more than twice the minimum biomass level
indicating highly eutrophic water. Samples were primarily phosphorus-limited,
but sometimes the reservoir was limited so closely by both nitrogen and phos-
phorus that the primary limiting nutrient could not be determined. Samples
collected after an underflow destroyed thermal and chemical stratification and
induced extensive vertical mixing would not support algal growth in control or
EDTA spiked samples. Indigenous phytoplankton contained approximately half
the normal chlorophyll concentration / unit volume. Correlation was high
between indigenous phytoplankton and S. capricornutum dry weight if considera-
tion was given to whether the reservoir was stratified or homothermal. An
equation was developed to predict mean indigenous phytoplankton standing
48
-------
crops. If the reservoir is homothermal, the equation is useful to determine
approximate indigenous phytoplankton standing crop at the time of sampling.
If stratified, it indicates the phytoplankton standing crop which was present
approximately two weeks earlier. Another equation was developed to predict
chlorophyll concentrations. (Abstract-Buchanan-Davidson-Wisconsin WRSIC).
0047
Haaland, P. T. , and G. Knutson. 1973. Growth experiments with Selenastrum
capricornutum Printz, p. 69-72. In Algal assays in water pollution
research, Proc. Nordic Symp., 25-26 October 1972, Oslo, Norway.
NORDFORSK, Secretariat of Environmental Sciences.
algal assay, temperature, light, phosphate, algae, Selenastrum capricornutum
Printz, growth physiology.
The report is of a study initiated for reasons of acquiring knowledge
about the growth physiology of the alga Selenastrum and to gain experience so
as to design and improve general, applicable tests; also, to search for extra
and possibly more suitable test algae. Growth as a function of temperature
and light were measured; investigations of growth in dilute medium and growth
in phosphate deficient conditions were discussed.
The authors concluded that use of Selenastrum in an algal test gave
reliable results within 20 hours when increased cell numbers were indications
of growth. (Abstract-Leischman-EPA).
0048
Hall, R. H. 1973. An algal toxicity test used in the safety assessment of
detergent compounds. Presented at 36th Annual ASLO meeting, 12 June
1973, Salt Lake City, Utah.
algal assay, bottle test, algae, Selenastrum capricornutum, Navicula semi-
nulum, Microcystis aeruginosa, toxicity, algistatic, detergents, algicidal,
inhibition, diatoms.
Detergent components which can enter surface waters are screened for
their environmental safety. An important part of this effort involves expo-
sure of aquatic organisms to selected levels of these compounds. A standard
algal toxicity test for use in this assessment is not now available. A pro-
cedure for the determination of the relative toxicity of detergent components
to phytoplankton has proven useful and informative. A five-day exposure of
49
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Selenastrum capricornutum, Microcystis aeruginosa, and Navicula seminulum to
levels of test materials ranging from 0.1 to 1000 mg/1 is followed by a nine-
day recovery period in the absence of the compound. ][n vivo fluorescence
measurements and cell population counts, made during the 14 days, allow quan-
titation of inhibition ranging from reduced growth to algistatic to algicidal
responses. The 5-day Minimum Algistatic Concentration (MAC-5 day) of a deter-
gent component is considered as potentially harmful to the phytoplankton
community. An important part of the test interpretation involves data evalu-
ation in light of potential and actual detergent component levels in surface
waters. Test results on specific surfactants and builders are presented which
illustrate the usefulness of these procedures in predicting the environmental
safety of detergent components. (Abstract-Author).
0049
Hendricks, A. C. 1978. Response of Selenastrum capricornutum to zinc sul-
fides. J. Water Pollut. Control Fed. 50(1):163-168.
algae, Selenastrum capricornutum, continuous flow, algal assay, toxicity,
heavy metals, zinc, sulfides, oxygen.
Oxygen production by Selenastrum capricornutum was determined after
exposing the cells to various concentrations of ionic zinc, sulfide, and zinc
plus sulfide. One mg/1 and 2.5 mg/1 of Zn2+ and S2- reduced oxygen production
by 50 percent or more. However, the two exhibited antagonistic behavior when
tested together. When 5 mg/1 of Zn2+ was mixed with 2.5 mg/1 of S2- only a
slight reduction in oxygen production occurred. (Abstract-JWPCF).
0050
Hostetter, H. P. 1976. Rapid bioassay for algal toxins in effluents and
surface waters. Ohio State Univ. RF NO 3934, U.S. Environmental Protec-
tion Agency, Cincinnati, Ohio, Final Report. 41 p.
algal assay, algae, Selenastrum capricornutum, pollution, nutrients, toxicity,
DO, oxygen electrode, limiting nutrients, copper, mercury, zinc, agricultural
watersheds, heavy metals.
A preliminary technique has been developed for a rapid bioassay to mea-
sure the effects of pollutants or nutrients on the metabolism of algae. A
Clark type oxygen electrode is used to measure changes in dissolved oxygen in
50
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a photosynthetic chamber containing the water to be tested and the test organ-
ism. The test alga is incubated 24 hr in the water prior to the test. Deter-
mination of photosynthetic rate takes 10-15 min. Evaluation of the water can
be made the day following collection. Results of the rapid bioassay are
compared with the Algal Assay Procedure Bottle Test. In culture medium the
rate of photosynthesis is directly proportional to the concentration of nutri-
ents limiting photosynthesis. Photosynthesis in waters collected from agri-
cultural watersheds (expected to be high in nitrogen) is not enhanced or only
slightly enhanced by fortification with algal growth medium or nitrate.
Photosynthesis of cells incubated in waters with toxic concentrations of cop-
per, mercury, or zinc is inhibited. (Abstract-Author).
0050-a
lonescu, A., and L. Gavrila. 1972. Contribution to study of the influence of
some stimulatory and inhibitory substances on growth and photosynthesis
on algae. Stud. Cercet. Biol. Sci. Bot. 24(1):9-16. In French.
algae, inhibition, indolebutyric acid, colchicine, atrazine, chlorelline
extracts, scenedesmine, diatoms, Scenedesmus, Chlorella, Selenastrum,
Navicula, Cocconeis, Amphora, Cymbella, Gomphonema, algal assay.
In research designed to investigate the multiplication, growth, and
intensity of photosynthesis in algae in pure or combined cultures, different
doses of IAA, indolebutyric acid, colchicine, atrazine, and extracts of chlor-
elline and scenedesmine were administered to cultures of Scenedesmus,
Chlorella, and Selenastrum. Phytoplankton (diatoms-predominantly Cocconeis,
Navicula, Amphora, Cymbella, Gomphonema) were also tested. The alga cultures
responded favorably to the action of the growth factors even during the first
few days of application, while colchicine and extracts of old cultures slowed
cellular multiplication. Photosynthesis was inhibited by colchicine and
atrazine and favored by the growth substances. In stopping photosynthesis,
atrazine revealed the existence of a heterotrophism in Scenedesmus that gave
it the opportunity to live without chlorophyll assimilation. Also, certain
combinations of culture media and growth substances removed the retarding
effect on multiplication and photosynthesis in old cultures and allowed an
increase in biomass. (Translated Summary-Biological Abstracts, Inc.).
51
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Q051
Jadlocki, J. F., Jr., J. Saldick, S. E. Coleridge, W. W. Smith, J. W. Brown,
and C. S. Nicholson. 1976. Effects of water hardness, phosphorus con-
centration and sample pretreatment of the Algal Assay Procedure Bottle
Test, p. 323-334. In E. J. Middlebrooks, D. H. Falkenborg, and T. E.
Maloney [eds], Biostimulation and nutrient assessment, Ann Arbor Sci.,
Mich.
algal assay, bottle test, algae, Selenastrum capricornutum, phosphorus, pre-
treatment, lake waters, water hardness.
To determine if different types of pretreatment affect the results of the
Algal Assay Procedure Bottle Test, chemical and algal assays were made on
samples from lakes in New York and Indiana. Algal assays using Selenastrum
capricornutum were done on water samples before treatment or after pretreat-
ment by autoclaving and/or filtration. The results indicated that pretreat-
ment of the water samples may cause erroneous results in the Algal Assay
Procedure Bottle Test. The controlling factors include water hardness, phos-
phorus concentration, and standing algal crops. Waters which are very hard
(above 200 ppm as calcium carbonate) and have a high orthophosphate concen-
tration (above 10 micrograms/L) may form a precipitate during autoclaving
which effectively depletes the available phosphorus, especially if the pre-
cipitate is removed by filtration. A substantial growth of algae (over 200
fluorescence units from J_n vivo chlorophyll a) in these waters would release
enough dissolved phosphorus during autoclaving to cause a net increase in
soluble phosphorus concentration. Much of the increased analyzable orthophos-
phate was apparently not available for Selenastrum growth. Since the expected
growth corresponding to the increased orthophosphate analyses was not ob-
served, filtration pretreatment may also remove large amounts of total phos-
phorus from eutrophic water samples. (Abstract-Buchanan-Davidson-Wisconsin
WRSIC).
0052
Joint Industry/Government Task Force on Eutrophication. 1969. Provisional
algal assay procedure. Joint Indust./Govt. Task Force Eutroph. , Grand
Central Station, New York. 62 p.
algal assay, algae, Selenastrum capricornutum, Anabaena flos-aquae. Micro-
cystis aeruginosa, continuous flow, bottle test, nutrients, eutrophication,
pollution, nitrogen fixation, carbon-14.
52
-------
The procedure described was developed by a team of international experts
/-
under the sponsorship of the Joint Industry/Government Task Force on Eutro-
phication and represents the first step toward developing reliable and repro-
ducible tests to determine the capacity of various aquatic environments to
grow algae. It is emphasized that the procedures presented are provisional
and are not standardized tests. Three procedures are described: a bottle
test, a continuous-flow chemostat test, and an jm situ test. Detailed de-
scriptions of equipment required, laboratory procedures, analytical tech-
niques, and sample preparation are given. Indicator organisms selected for
use in the tests are Selenastrum capricornutum, a green alga, Anabaena flos-
aquae, a blue-green alga capable of fixing nitrogen; and Microcystis
aeruginosa a blue-green alga not capable of fixing nitrogen. (Abstract-
Uttermark-Wisconsin WRSIC).
0052-a
Justice, C. , S. Murray, P. S. Dixon, and J. Scherfig. 1972. Evaluation of
materials for use in algal culture systems. Hydrobiol. 40(2):215-221.
bottle test, continuous flow, algal assay, algae, Selenastrum capricornutum.
The report is of experiments conducted to test new materials used in
continuous flow algal assays. The materials tested were Pyrex and Kimax glass
flasks; neoprine stoppers and Cafe-au-lait rubber stoppers. The authors
concluded that new materials used in tests, as new, had some inhibitory ef-
fects on algal growth and that the new materials should be soaked prior to
use. The authors support testing the new materials before incorporating the
materials into important algal assays. (Abstract-Leischman-EPA).
0053
Kallqvist, T. 1973. Algal assay procedure (bottle test) at the Norwegian
Institute for Water Research, p. 5-18. IJTI Algal assays in water pollu-
tion research, Proc. Nordic Symp., 25-26 October 1972, Oslo, Norway.
NORDFORSK, Secretariat of Environmental Sciences.
algal assay, bottle test, limiting nutrients, algae, Selenastrum capricorn-
utum, Phaeodactylum tricornutum, Skeletonema costatum, toxicity, PCB, phos-
phorus, wastewater, diatoms.
53
-------
The report is an outlined description of the laboratory procedure using
the algal assay bottle test to determine the following: (1) the toxicity of
substances on growth of algae; (2) the maximum standing crop in dilute medium;
(3) the effect of phosphorus on the maximum standing crop; (4) sewage effluent
effects on algal growth; and (5) determination of limiting nutrients. The
algal species used were Selenastrum capricornutum, Skeletonema costatum and
Phaeodactylum tricornutum. Brief examples of results are given, graphs in-
cluded, but no discussion is provided. (Abstract-Leischman-EPA).
0054
Kallqvist, T. 1973. Use of algal assay for investigating a brackish water
area, p. 111-124. Jji algal assays in water pollution research, Proc.
Nordic Symp., 25-26 October 1972, Oslo, Norway. NORDFORSK, Secretariat
of Environmental Sciences.
wastewater, nitrogen, phosphorus, N:P ratios, rivers, limiting nutrients,
algae, Selenastrum capricornutum, Phaeodactylum tricornutum, Skeletonema
costatum, fjord, algal assay, bottle test, diatoms.
The algal assay procedure was used to investigate the Frierfjord and
influences of wastewater upon the nutrient status of the fjord. The algal
species used were Selenastrum capricornutum, Phaeodactylum tricornutum,
Skeletonema costatum. The author found that the wastewater had a stimulatory
effect on the growth of algae in the fjord as well as growth further out from
the coastal area. Phosphate was found to be the primary limiting nutrient. A
high N:P ratio was calculated for the wastewater. The author concluded that
the algal assay procedure was advantageous in that it expressed clearly the
characterizations of waters in terms of nutrient status more than would chemi-
cal analyses. (Abstract-Leischman-EPA).
0055
Kallqvist, T. 1974. Algal growth potential of six Norwegian waters receiving
primary, secondary, and tertiary sewage effluents. Ver. Int. Assoc.
Theor. Appl. Limnol. Proc. 19(3):2070-2081.
AGP, wastewater, lake waters, fjord, algae, Selenastrum capricornutum, Phaeo-
dactylum tricornutum, phosphorus, nitrogen, limiting nutrients, algal assay,'
diatoms.
54
-------
The experiments with algal growth in mixtures of three different kinds of
treated sewage and six receiving waters have shown that the growth potential
in the mixture depends on the composition of the receiving water as well as
the wastewater. This is especially the case for chemically treated wastewater
(CTW). With this kind of effluent, the P concentration in the receiving water
is important for the resulting growth potential. In P deficient waters the
fertilizing effect is very low, but in waters having P in excess as is usually
the case in sea water, low concentrations of chemically treated sewage may
give the same increase as biologically treated sewage (BTW). In such waters
treatment methods that reduce N would be the best solution (Goldman et al,
1974). The reason why an addition of CTW to P deficient waters causes almost
no increase in growth potential is that the P content in the sewage after the
chemical precipitation is low. It must be pointed out, however, that the
assays were carried out on filtered water, which means that the P adsorbed to
the particulate matter was removed more efficiently than in most treatment
plants. Later experiments have shown that much of this particulate-bound P is
readily available to the algae and consequently that algal assays on filtered
water from a chemical treatment plant give values which are too low (Lofgren,
1974). This finding emphasizes the point that filtering of the effluents from
the chemical treatment plants will bring about better P removal.
Of the mechanically and biologically treated wastewaters, the MTW con-
tains the higher level of nutrients and also gives a greater increase of the
growth potential in all receiving waters. The difference between these two
types of wastewater regarding their fertilizing effect is, however, small.
BTW seems to have a more immediate stimulating effect on the algal growth than
MTW, resulting in a higher growth rate of the test algae in cultures with BTW.
The reason for this has not been investigated. (Conclusion-Author).
0056
Katko, A. 1975. Algal assays for the National Eutrophication Survey, p.
44-52. In Proc. Biostim. Nutr. Assess. Workshop, 16-17 October 1973,
U.S. Environmental Protection Agency, Corvallis, Oregon. EPA-660/3-75-
034.
algal assay, lake waters, bottle test, algae, Selenastrum capricornutum,
phosphate, nitrogen, wastewater.
55
-------
The algal assay procedure was incorporated into the National Eutrophi-
cation Lake Survey. The investigation was of the nutrient status (primarily
nutrient reduction to improve water quality) of approximately 800 lakes in the
United States. The report is of the programs completed and those in progress,
for all 27 states east of the Mississippi River. The programs involved 485
lakes and 2500 stream sampling sites. Selenastrum capricornutum was the test
alga used for the algal assay. Results are given for the 1972 sampling year.
A discussion is given of the algal assay as a tool; and, the advantages and
disadvantages in relating the results of algal assays to real world situations
in the attempt to assess and classify all the lakes within the Lake Survey.
(Abstract-Leischman-EPA).
0057
Keenan, J. D. , and M. T. Auer. 1974. The influence of phosphorus luxury
uptake on algal bioassays. J. Water Pollut. Control Fed. 46(3):532-542.
algal assay., phosphorus, uptake, bottle test, algae, Anabaena flos-aquae,
Microcystis aeruginosa, Selenastrum capricornutum.
The effect of luxury uptake of orthophosphate on the sensitivity of algal
growth bioassays was determined. Short-term bioassays consisted of gravimet-
rically determined harvested dry weights; long-term bioassays consisted of
manometrically determined photosynthetic rates. Phosphorus metabolism was
studied by measurement of stored phosphorus, alkaline phosphatase activity,
and phosphate concentration in the nutrient medium. Anabaena flos-aquae,
Microcystic aeruginosa, and Selenastrum capricornutum exhibited luxury uptake
of phosphorus. However, M. aeruginosa and S. capricornutum did not exhibit
alkaline phosphatase activity. It was concluded that short-term bioassays are
less sensitive to phosphorus concentration than long-term bioassays. The
influence of luxury uptake is a function of the phosphorus concentration in
the original growth medium and of the length of the starvation period. (Ab-
stract- JWPCF).
0058
King, D. L. , and J. T. Novak. 1974. The kinetics of inorganic carbon-limited
algal growth. J. Water Pollut. Control Fed. 46(7):1812-1816.
56
-------
inorganics, carbon, algae, Selenastrum capricornutum, Scenedesmus quadricauda,
growth rates, algal assay.
The data of an earlier paper by J. C. Goldman and others is reinterpreted
to show that Selenastrum capricornutum and Scenedesmus quadricauda both re-
spond in a kinetic fashion to free C02 concentration of water for their photo-
synthetic C source, and that their specific growth rate is not related di-
rectly to total inorganic C in the water as suggested by Goldman et al.
(Abstract-Biological Abstracts, Inc.).
0059
Klotz, R. L. , J. R. Cain, and F- R. Trainor. 1975. A sensitive algal assay:
An improved method for analysis of freshwaters. J. Phycol. 11(4):411-
414.
algal assay, algae, Selenastrum capricornutum Printz, rivers, bottle test.
An algal assay capable of detecting the carrying capacity of nutrient-
poor rivers was developed. In order to eliminate carry-over of nutrients,
thus masking the true nutrient condition of the water tested, a new growth
medium was developed. This maintenance medium contained less than 4 mg-1-1
total inorganic salts. With an inoculum of 104 cells ml-1 the test organism
Selenastrum capricornutum Printz was grown in sterile-filtered river water and
transferred daily with a 1:1 dilution. The assay was carried out for 4 or 5
days, at which time cell number was calculated, and the doublings per day were
determined. Comparisons are made with similar but less sensitive assays, on
the basis of both daily and 5-day readings. (Summary-Author).
0059-a
Kotai, J. , T. Krogh, and 0. M. Skulberg. 1978. The fertility of some
Norwegian inland waters assayed by algal cultures. Mitt. Int. Ver.
Limnol. 21:413-436.
eutrophication, lake waters, rivers, nutrients, bottle test, algal assay,
algae, Selenastrum capricornutum Printz, nitrogen, phosphorus, AGP.
In an investigation and comparative study of more than 400 lakes and
rivers in East Norway, algal assays were conducted to determine algal growth
potential values; the limiting nutrients; and, the fertility of the waters
57
-------
examined. The algal assays were found to provide more information about
biostimulatory and bioinhibiting properties of the waters than the chemical
analyses. The authors felt that both chemical analysis and algal assays
should be combined in studies of polluted waters. (Abstract-Leischmn-EPA).
0060
Kotai, J. , T. Krogh, K. Ormerod, and 0. M. Skulberg. 1976. Estimation of
algal growth potential. NIVA Pub!., D2-25, Oslo, Norway. 32 p.
AGP, algae, Selenastrum capricornutum, Phaeodactylum tricornutum, algal assay,
diatoms.
The report is a detailed, outlined procedure for use and application of
the algal growth potential, AGP, as a method to test a laboratory sample under
set standard conditions, either freshwater or marine water. The authors
provide detailed descriptions of the necessary, reagents, apparatus, test
organisms, and methods to conduct an experiment utilizing the AGP method. The
test organisms suggested are Selenastrum capricornutum and Phaeodactylum
tricornutum. (Abstract-Leischman-EPA).
0060-a
Laake, M. 1978. Monitoring the effects of chemical and biological waste
water treatment i_n situ by dialysis cultures of freshwater algae. Mitt.
Int. Ver. Limnol. 21:453-472.
continuous flow, algal assay, algae, dialysis, chlorophyll a, growth kinetics,
ATP, heavy metals, zinc, copper, lead, cadmium, Scenedesmus quadricauda,
Selenastrum capricornutum, Chlamydomonas reinhardtii, Asterionella formosa,
diatoms.
Unialgal and mixed cultures of freshwater algae were inoculated in 12
dialysis bags (32 ml) attached to a rotating wheel submerged in throughflow
boxes of white polyethylene. The surface was freely exposed to daylight. Two
parallels were harvested every third or fourth day and the growth followed by
measuring optical density at 430 mm. The cells were analyzed for organic
carbon, chlorophyll a and phaeophytin, ATP and the heavy metals Cu, Zn, Cd and
Pb. Four experiments each with four types of treated waste water at 0.5 and 5
vol % concentrations were conducted: secondary Al-treatment + biological
oxidation pond (Al + BD), secondary Al-treatment (Al), activated sludge bio-
58
-------
logical treatment + Al-treatment (B + Al) and activated sludge with simultan-
eous Al-treatment (SIM). Two experiments with Scenedesmus quadricauda and two
starting with equal numbers of Scenedesmus, Selenastrum capricornutum Chlamy-
domonas reinhardtii and Asterionella formosa were performed.
The results show that SIM generally gave highest cell yield, while the
others were more or less equal. With mixed cultures the ATP content per cell
decreased with decreasing waste water load, but increased with Scenedesmus.
Mean levels of 0.6 pg ATP/mg C confirm phosphate limitation. Chlorophyll
content significantly decreased with decreasing load, while no differences
between treatments were evident. Chlamydomonas and Selenastrum did grow to a
lesser extent at 5% than at 0.5% dosing, while Asterionella diminished almost
completely. All mixed cultures were dominated by Scenedesmus, which appeared
to grow better than in pure culture.
The yield factors to phosphate and the maintenance rates were calculated
from growth rate in linear phase and asymptotic biomass, respectively. These
factors gave the best basis for comparative evaluation of the treatment pro-
cesses. The results show that the algal growth efficiency on phosphate limi-
tation may be ranged in the order Al + BD, Al, B + Al and SIM, thus indicating
the degree of environmental stress in the cultures. The efficiency also
decreased with increasing load for all treatments.
From data on heavy metal content concentration factors in the order of
magnitude 106 for Cd, 4-105 for Pb and 2.5-104 for Zn were calculated. High
levels were correlated with high maintenance rates versus phosphate. (Sum-
mary-Author).
0060-b
Lambou, V. W. , L. R. Williams, S. C. Hern, R. W. Thomas, and J. D. Bliss.
1976. Prediction of phytoplankton productivity in lakes, p. 696-700. I_n
Proc. Conf. Environ. Modeling and Simulation, U.S. Environmental Protec-
tion Agency, 19-22 April 1976, Cincinnati, Ohio. EPA-600/9-76-016.
algal assay, bottle test, algae, chlorophyll a, water quality, phytoplankton,
lake waters, limiting nutrients, phosphorus, nitrogen, N:P ratios, Selenastrum
capricornutum, Anabaena, Microcystis.
This study presents relationships between phytoplankton productivity as
measured by yearly mean chlorophyll a levels, and ambient water quality and
hydrologic measurements. Among the nutrients examined, phosphorus forms were
59
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most highly correlated with chlorophyll a levels. The effects of such factors
as retention time, primary nutrient limitation, stratification, and macrophyte
dominance upon productivity responses are evaluated. Additional parameters
related to productivity include turbidity, Secchi disc, nitrogen to phosphorus
ratio, pH, total alkalinity, and forms of inorganic nitrogen. Discussions of
the factors affecting phytoplankton productivity and the application of the
limiting nutrient concept are included. (Summary-Author).
0061
Lange, W. 1971. Limiting nutrient elements in filtered Lake Erie water.
Water Res. 5(1):1031-1048.
algal assay, lake waters, limiting nutrients, nitrate, phosphorus, cobalt,
iron, algae, Anabaena circinalis, Microcystis aeruginosa, Nostoc muscorum,
Selenastrum capricornutum. bottle test.
Biweekly laboratory bioassays were made using filtered Lake Erie waters
frdm the Bass Island region to simulate the effect of an influx of individual
nutrients to Lake Erie water. Each sample during the 1969 growing season was
inoculated singly with each of three blue-green algae and one green algal
species. Portions of each inoculated water sample were enriched with one of
16 essential nutrient elements and cell numbers were determined in the fourth
week after exposure under controlled culture conditions. Comparative cell
counts showed nitrate-N was the nutrient the most frequently required and was
limiting for these algal species in about 2/3 of the water samples. Phosphor-
us, cobalt and chelated iron were limiting for about 1/3 of their cultures.
To achieve maximum growth stimulation generally required a combination of
several nutrient elements. An Aphanizomenon bloom occurred naturally in Lake
Erie at the water sampling site and after its collapse from nutrient insuffi-
ciency, the water samples were found to be toxic to certain of the test algal
species. (Abstract-Author).
0062
Lee, C. C. , R. F. Harris, J. K. Syers, and D. E. Armstrong. 1971. Adenosine
triphosphate content of Selenastrum capricornutum. Appl. Microbiol.
21(5):957-958.
ATP, adenosine triphosphate, phosphorus, algal assay, algae, Selenastrum
capricornutum.
60
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The adenosine triphosphate content of Selenastrum caprlcornutum was
maintained within 1.4 to 3.4 |jg of adenosine triphosphate/mg (dry weight) of
living biomass at different growth stages in media of different phosphorus
concentrations. (Abstract-Author).
0063
Lehmusluoto, P. 0. 1973. Algal assay procedure in use in Finland, p. 33-34.
Jj} Algal assays in water pollution research, Proc. Nordic Symp. , 25-26
October 1972, Oslo, Norway. NORDFORSK, Secretariat of Environmental
Sciences.
algal assay, bottle test, nitrogen, phosphorus, algae, Ankistrodesmus
falcatus, Selenastrum capricornutum, Chlorella sp. , wastewater.
The report is a description of an algal assay procedure, the AGP (Algal
Growth Potential), used in Finland to investigate the nutrient status of
various types of waters. The test organisms described as those being used are
Ankistrodesmus falcatus, Selenastrum capricornutum, and Chlorella sp. Brief
explanations of steps in the procedure and references are given. (Abstract-
Lei schman-EPA).
0064
Lindemann, E. G. , and G. P. Volpp. 1971. Carbon dioxide and trace elements
as factors in algal assay procedures. Presented Water, Air, and Waste
Chem., Amer. Chem. Soc., Washington, D.C. 12 p.
algal assay, bottle test, carbon dioxide, algae, Selenastrum capricornutum,
trace elements.
The report is a discussion of the investigation on the effect of bac-
teria-generated carbon dioxide on the growth of the green alga Selenastrum
capricornutum and the final yield of the algae under laboratory conditions.
The second portion of the report covers the importance of trace elements and
the required precautions in identifying them as limiting nutrients. The
authors found that the maximum cell count was more than three times greater in
the flasks supplied with bacteria-generated C02 as opposed to the flasks with
C02 supplied from the atmosphere. The authors suggested the use of polycar-
bonate flasks for the procedure sensitive to trace elements and the technique
described in the report. (Abstract-Leischman-EPA).
61
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0065
Lindmark, G. 1973. Bioassay with Selenastrum capn'cornutum to assess the
nutrient status of lakes and the fertilizing influence of interstitial
water, p. 73-80. Ir\ Algal assays in water pollution research, Proc.
Nordic Symp., 25-26 October 1972, Oslo, Norway. NORDFORSK, Secretariat
of Environmental Sciences.
lake waters, algal assay, algae, Selenastrum capricornutum, wastewater, eutro-
phication, restoration.
The algal assay method, as a form of investigation of the nutrient status
of lake waters, was incorporated into the Lake Trummen Restoration Project.
The study used the algal species Selenastrum capricornutum. The restoration
process was by removal of lake sediments. The algal assay was used as a tool
to study the nutrient status of interstitial waters and lake waters before,
during, and after restoration. The author found a marked seasonal variation
in the nutrient levels; phosphorus was always in excess: additions of the
interstitial water to the lake increased heavy algal growth. (Abstract-
Lei schman-EPA).
0065-a
Little, L. W. , and M. A. Chillingworth. 1974. Effect of 56 selected dyes on
growth of the green alga Selenastrum capricornutum. Chap. 2. Ln Dyes
and the environment - Reports on selected dyes and their effects, v. II,
Amer. Dye Manuf. Inst. , Inc., New York.
dyes, toxicity, inhibition, algal assay, algae, Selenastrum capricornutum,
fish, Pimephales promelas, bottle test.
Fifty-six commercially important dyes, selected by American Dye Manufac-
turers Institute Ecology Committee, were tested for toxicity to the green alga
Selenastrum capricornutum. The Algal Assay Procedure developed by the Na-
tional Eutrophication Research Program—Environmental Protection Agency was
employed. Concentrations of 1 and 10 mg/1 of dye were assayed. Growth of
algae was measured by direct counts at 7 and 14 days of incubation.
In general, basic dyes, even at a concentration of 1 mg/1, inhibited
algal growth to less than 1% of that attained in the dye-free controls. Dyes
of other classes were generally less toxic. Overall, the effect of dyes on
algal growth correlates well with that of the same dyes on the fathead minnow,
Pimephales promelas. (Abstract-Author).
62
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0066
Little, L. , and D. Pittman. 1974. Determination of herbicides and their
relationships to algal growth. Proc. Nebr. Acad. Sci. Affil. Soc. 84:48.
herbicides, dacthal, aldrin, algal blooms, lake waters, algae, Selenastrum,
phytoplankton, Anacystis, Pediastrum, algal assay.
Herbicide concentrations, with emphasis on dacthal, and their relation-
ships to algal growth was studied in connection to algal blooms. Dacthal was
examined as a possible stimulant. Anacystis was stimulated by a concentration
of 0.01 ppm dacthal, but Pediastrum and Selenastrum results were inconclusive.
Algae, plankton, water, and soil samples from Bluestem Lake near Lincoln,
Nebraska were found to contain pesticides, tentatively aldrin and dacthal.
Extraction, cleanup, and analysis methods for lake samples outlined. Lake
algal blooms are not, conclusively, due to the stimulating effects of herbi-
cides. (Abstract Only-Author).
0066-a
Long, E. T., and G. D. Cooke. 1978. Phosphorus variability in three streams
during storm events: chemical analysis vs. algal assay. Mitt. Int. Ver.
Limnol. 21:441-452.
eutrophication, streams, phosphorus, lake waters, algal assay, bottle test,
algae, Selenastrum capricornutum, urban runoff, forest runoff.
The majority of calculations of P loading to lakes have been made on the
basis of a chemical analysis. The present study was conducted to evaluate and
compare chemically determined total phosphorus (TP) and soluble reactive
phosphorus (SRP) to biologically available phosphorus (AP) loading of lakes
during hydro!ogical events (storm runoff) and calendar events (weekly sam-
ples).
Three streams draining a forest, an urban region, and a lake, all located
in the Twin Lakes Watershed (NE Ohio, U.S.A.) were studied. Water samples
were collected throughout the course of storm events. They were also collec-
ted at weekly intervals for a 51 day period prior to storm sampling. Measure-
ment of stream flow, suspended solids, soluble reactive and total P, amount of
precipitation (rain), and storm intensity were taken. A portion of each water
sample was assayed for biologically available P using a modified Algal Assay
Procedure with Selenastrum capricornutum Printz.
63
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Changes in loading of the three forms of P, calculated from weekly sampl-
ing, are depicted.
The response of the urban and forest area to two storm events was one of
increased water loading. The response of the lake outflow to storm initiation
was increased water loading, but this stream did not display the abrupt ampli-
tude changes that were exhibited by the other two streams. Changes in sus-
pended solids loading during storm events is depicted. The changes in TP, SRP
and AP loadings are also illustrated.
Comparison of loading rates from calendar events to those from hydro-
logical events showed that considerably greater amounts of all three forms of
P were contributed during storm runoff from all three areas. The urban area,
however, contributed substantially larger amounts of all three P forms than
did the other two areas.
The conclusions reached from this study were that short-term phenomena,
such as storm runoff, do play a major role in P loading to lakes. It was also
shown that land use activities greatly affect these loading rates and that
urban areas have the greatest impact. Available phosphorus (AP) loadings were
considerably different than total phosphorus (TP) loading, both during storm
events and during base flow. During base flow, only a small fraction of the P
income was AP, this fraction increased greatly during storm flow from the
urban and forest areas, but not from lake discharge. The suggestion is made
that the use of biologically available P in nutrient budgets might greatly
enhance the predictability of limnological models relating change in P income
to the degree of eutrophication. (Modified Summary-Author).
0067
Maki, A. W., L. D. Geissel, and H. E. Johnson. 1975. Toxicity of the lamp-
ricide 3-trifluoromethyl-4-nitrophenol to 10 species of algae, p. 3-17.
IP U.S. FishWildl. Serv. Invest. Fish. Control 56.
algae, Scenedesmus quadricauda, Stigeoclonium tenue, Selenastrum capricorn-
utum, Chlorella pyrenoidosa, Cylindrospermum sp. , Anabaena flos-aquae,
Anabaena cylindrica, Nostoc linokia, Nitzschia sp. , Navicula pelliculosa,
growth inhibition, TFM, algal assay, diatoms.
The toxicity of analytical and field grades of the lampricide 3-tri-
fluoromethyl-4-nitrophenol (TFM) to unialgal cultures of four green algae,
four blue-green algae, and two species of diatoms was examined in 96-h toxi-
64
-------
city tests. Growth was measured by daily optical density readings, cell
counts of nonfilamentous species, and a gravimetric determination of maximum
standing crop at the end of the tests. A 50% inhibition of growth (calculated
from optical density data) occurred at concentrations less than 10 mg/1 of TFM
for all species tested except Chlorella pyrenoidosa, which was resistant at
concentrations greater than 15 mg/1. Inhibition of growth was lowest in the
blue-green species (50% effect levels, 9-10 mg/1), generally intermediate in
the green algae, and severe in diatoms (50% effect levels, 1-4 mg/1). Field
grade TFM tended to be more toxic than the analytical grade. Similar toxicity
relationships were indicated by each of the three types of measurements made.
Growth tests conducted with high concentrations of TFM and subsequent
filtration of the cells and resuspension in toxicant-free medium indicated
that exposure to TFM at concentrations of 30 mg/1 for 96 h did not destroy the
viability of algal cells but temporarily inhibited growth. (Abstract-Author).
0068
Malone, R. F. , K. A. Voos, W. J. Grenney, and J. H. Reynolds. 1976. The
effects of media modifications upon Selenastrum capricornutum in batch
cultures,
-------
pH. It was suggested that a buffer be used in conjunction with sodium bicar-
bonate. (Abstract-Leischman-EPA).
0069
Maloney., T. E. , and W. E. Miller. 1975. Algal assays: Development and
application, p. 344-355. J.n Water quality parameters, Amer. Soc. Test.
Mater., ASTM STP 573.
water quality, limiting nutrients, algal assay, bottle test, wastewater, lake
waters, streams, algae, Selenastrum capricornutum.
The development, refinement, standardization, and practical application
of an algal procedure are described. Results of experiments are presented to
illustrate the use of the algal assay procedure to determine algal growth-
limiting nutrients in fresh waters, including lakes and streams; for the as-
sessment of receiving waters to determine their nutrient status and sensi-
tivity to change; for the assessment of the effects of changes in waste treat-
ment processes on receiving waters; and for the evaluation of materials and
products to determine their potential effects on algal growth. (Abstract-
Author).
0070
Maloney, T. E., W. E. Miller, and N. L. Blind. 1973. Use of algal assays in
studying eutrophication problems, p. 205-214. In S. H. Jenkins [ed.],
Advances in water pollution research, Sixth Internat. Conf., Jerusalem,
Pergamon Press, Oxford and New York.
algal assay, eutrophication, wastewater, lake waters, phosphorus, nitrogen,
nutrients, limiting nutrients, algae, Selenastrum capricornutum.
The use of algal assays to assist in the solution of practical eutroph-
ication is illustrated by the studies described herein. We have demonstrated
their use in assessing the effects of changes in waste treatement processes on
receiving waters, in identifying algal growth-limiting nutrients, and in
assessing receiving waters to determine their nutrient status and sensitivity
to change. We feel that algal assays will also be a valuable asset in dealing
with other practical problems such as evaluating materials and products to
determine their potential effects on algal growth in receiving waters.
66
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The algal assay cannot, of course, be used by itself in evaluating and
solving eutrophication problems. If combined, however, with good chemical and
physical analyses as well as thorough limnological investigations, it can be
an extremely useful procedure. (Summary-Author).
0071
Maloney, T. E. , W. E, Miller, and T. Shiroyama. 1972. Algal responses to
nutrient additions in natural waters: I. Laboratory Assays, p. 134-140.
_In G. F. Likens [ed.], Nutrients and eutrophication: The limiting nu-
trient controversy, Spec. Symp. Amer. Soc. Limnol. Oceanogr., Vol 1,
Allen Press, Kansas.
lake waters, nitrogen, phosphorus, carbon, algae, Selenastrum capricornutum
Printz, algal assay, bottle test.
Laboratory algal assays were conducted on waters from nine Oregon lakes
of varying water quality. Selenastrum capricornutum Printz was used as the
test species in an algal assay using the Provisional Algal Assay Procedure
(Joint Industry/Government Task Force on Eutrophication 1969). Additions of
nitrogen, phosphorus, and carbon, singly and in combination, were made to the
waters and algal growth rates were determined. The addition of phosphorus
alone greatly stimulated algal growth rates in four of the waters and the
addition of nitrogen alone slightly stimulated algal growth rates in waters of
two of the lakes. Three of the waters were capable of supporting relatively
high algal growth rates without nutrient additions, and in one highly oligo-
trophic lake water the addition of nitrogen, phosphorus, and carbon had no
effect on algal growth rates. In all cases algal growth rates were directly
proportional to the amounts of dissolved phosphorus in the waters, but there
was no obvious correlation between algal growth rates and concentrations of
nitrogen and carbon. (Abstract-Author).
0072
McDonald, G. C. , and N. L. Clesceri. 1973. Effect of wastewater organic
fractions on the growth of selected algae, p. 479-496. In E. Glass
[ed.], Bioassay techniques and environmental chemistry, Ann Arbor Sci.,
Mich.
wastewater, algal assay, carbon, phosphorus, organic fractions, bottle test,
algae, Anabaena flos-aquae, Selenastrum capricornutum.
67
-------
The existence and extent of algal growth enhancement brought about by the
addition of wastewater organic fractions to representative algal cultures were
determined. A sample of effluent from a conventional activated sludge facil-
ity located at Batavia, New York was subjected to fractionation using gel
chromatographic techniques. Subsequent to filtration and freeze drying, the
concentrated wastewater was separated into organic fractions by gel chroma-
tography. Algae selected for the study were Selenastrum capricornutum and
Anabaena flos-aquae. Growth of both species was determined by absorbance
measurements at 750 nm with a Beckman DU-2 Spectrophotometer. Organic com-
pounds contained in wastewater fractions have been found to exert a growth-
enhancing effect on the two algae species. The exact nature of the causative
growth factors and the pathway by which they achieve their effects are as yet
unknown, and further investigative efforts are being conducted to assess
generality of these results. This undertaking is important as increased algal
productivity may continue to occur if such factors are general constituents of
wastewater effluents, notwithstanding the removal of nitrogen and phosphorus
from such effluents. (Abstract-Jones-Wisconsin WRSIC).
0073
McDonald, G. C. , R. D. Spear, P. J. Lavin, and N. L. Clesceri. 1970. Kine-
tics of algal growth in austere media, p. 97-105. In J. E. Zajic [ed.],
Properties and products of algae symposium, Plenum Publ. Corp., New York.
nutrients, nitrogen, phosphorus, limiting nutrients, lake waters, inhibition,
growth kinetics, algae, Selenastrum capricornutum, media, algal assay.
The report is of experiments conducted to determine the effects of dif-
ferent concentrations of nitrogen and phosphorus on growth of the alga Sele-
nastrum capricornutum. Initially the use of the two media, modified Gorham's
medium and Basic ASM medium, diluted with glass filtered water, was part one
of the experiments. Secondly, the media were diluted with filtered Lake
George waters. Varied concentrations of nitrogen and phosphorus were added
and growth of the algae was measured. The authors found that the reduction of
nitrogen concentrations was less effective on reduction of algal growth than
the reduction of phosphorus concentrations for both experiments. The effect
of nitrogen reduction in the Lake George:media mixture was more pronounced
than in the distilled waterrmedia mixture. The alga was found to be phos-
68
-------
phorus limited; Lake George waters used as the dilutent had an inhibitory
effect on growth other than the limiting nutrients; and, phosphorus limitation
was found to be the least pronounced in the Lake George:Basic ASM medium
mixture. (Abstract-Leischman-EPA).
0074
Middlebrooks, E. J. , D. H. Falkenborg, and T. E. Maloney, editors. 1975.
Biostimulation and nutrient assessment. Ann Arbor Sci., 391 p.
algal assay, nutrients, eutrophication, toxicity, limiting nutrients, waste-
water, sediments, phosphorus, lake waters, bottle test, growth rates, indi-
genous phytoplankton, algae, Selenastrum capricornutum, Anabaena flos-aquae,
continuous flow, water hardness.
The application of the Algal Assay Procedure Bottle Test and its modifi-
cations are discussed in relation to various ecological and environmental
variables. Contributions focus on the application of the test in biostimu-
lation and toxicity testing; field evaluation of the test on surface waters of
North Carolina, its application to define the effects of wastewater effluents
upon algal growth in multiple use river systems; and the relationship of
laboratory algal assays to measurement of indigenous phytoplankton in Long
Lake, Washington. The test's applicability to compare algal growth responses
of Selenastrum capricornutum and Anabaena flos-aquae in waters collected from
Shagawa Lake, Minnesota, seasonal variation of algal biomass production poten-
tial and nutrient limitation in Yaquina Bay, Oregon; and algal nutritional
bioassays of Lake Wylie, North Carolina, is described. Nutrient assessment as
a basis for lake management priorities, as applied to Wisconsin lakes, is re-
viewed. Other contributions compare semi-continuous and continuous flow bio-
assays; the effects of media modifications upon S. capricornutum in batch cul-
tures; sediment-water microcosms designed for assessment of nutrient inter-
actions; effects of water hardness, phosphorus concentration and sample pre-
treatment on the test; and an elutriate method to determine the significance
of dredged material disposal in open waters. (Abstract-Auen-Wisconsin).
0075
Middlebrooks, E. J. , E. A. Pearson, M. Tunzi, A. Adinarayana, P. H. McGauhey,
and G. A. Rohlich. 1971. Eutrophication of surface water: Lake Tahoe.
J. Water Pollut. Control Fed. 43(2):242-251.
69
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ammonia, carbon, eutrophication, lake waters, nitrogen, nitrogen oxide, phos-
phorus, nutrients, streams, bottle test, algae, Selenastrum gracile, contin-
uous flow, algal assay.
Although nitrogen and phosphorus are major factors in the nutrient-rich
or eutrophic condition that leads to algal blooms, their presence is not
always the critical factor in eutrophication. The authors make an effort to
develop suitable methods of assaying the algal growth-stimulating potential of
various influents to lakes and streams and to apply these methods to an eval-
uation of their ability to increase the eutrophication of water. A test
algae, Selenastrum gracile, was used to assess quantitatively biostimulation
by exposure to surface runoff, seepages, effluents from various wastewater
treatment processes, and Lake Tahoe (Calif.) water. Initial growth response
to the test algae was much greater with batch-type assay data than with
steady-state conditions. Both batch and continuous flow assays indicated that
the biostimulatory properties of secondary effluents were much greater than
that from raw or primary treated wastes. Wastewater effluents of all types
showed higher growth response rates than could be accounted for by the amount
of N and P present in the samples. (Abstract-JWPCF).
0076
Miller, W. E. , and T. E. Maloney. 1971. Effects of secondary and tertiary
wastewater effluents on algal growth in a lake-river system. J. Water
Pollut. Control Fed. 43(12):2361-2365.
algal assay, eutrophication, nitrogen, nutrients, phosphorus, water quality,
wastewater, lake waters, rivers, algae, Selenastrum capricornutum, bottle
test.
The biostimulatory effects of secondary and tertiary treated wastewater
effluent were determined on the Shagawa Lake-Burntside River system in Minne-
sota. Chemical characteristics of the river were consistent year-round and
supported growth of test algae in concentration of 5 X 103 cells/ml. The
addition of 0.02 mg phosphorus (P)/l increased algal growth. The addition of
1.0 mg nitrogen (N)/l resulted in no increase. The addition of 2 percent
secondary effluent increased algal growth significantly. Ten percent tertiary
effluent did not stimulate growth. Lake water supported algal growth, and
correlation was shown between algal growth responses and N and P content of
70
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lake water samples. The lake was N and/or P deficient, depending on the time
of the year. (Abstract-JWPCF).
0077
Miller, W. E., J. C. Greene, and T. Shiroyama. 1976. Application of algal
assays to define the effect of wastewater effluents upon algal growth in
multiple use river systems, p. 77-92. lr\ E. J. Middlebrooks, D. H.
Falkenborg, and T. E. Maloney [eds.], Biostimulation and nutrient assess-
ment, Ann Arbor Sci., Mich.
wastewater, rivers, limiting nutrients, nitrogen, phosphorus, trace elements,
heavy metals, toxicity, N:P ratios, algal assay, bottle test, algae, Sele-
nastrum capricornutum.
The report is of application of algal assays to define effects of domes-
tic and industrial waste discharges on algal growth in the Spokane River
System, Washington. The design and interpretation of laboratory algal assays
is discussed. Algal responses to nitrogen and phosphorus limitations, trace
element deficiency, and heavy metal toxicity were determined. The critical
nitrogen:phosphorus ratio to support Selenastrum capricornutum was 11.3:1
micrograms. This ratio can assess algal growth limitations in natural waters.
Waters with N:P ratios less than 10 are considered nitrogen-limiting while
waters with N:P ratios above 10 may be phosphorus-limiting for algal growth.
Correlation of algal assays with yields predicted from N:P ratios depend on
the reliability of test procedure. Total soluble inorganic nitrogen (nitrite
+ nitrate + ammonia) and orthophosphate concentrations or availability in
samples were calculated by dividing maximum assay yields by the total soluble
inorganic nitrogen or orthophosphate yield coefficient. That samples do not
attain the predicted assay yield or nutrient limitation status may be due to
growth-1imiting nutrients other than nitrogen or phosphorus, presence of toxic
materials, or unreliable chemical determinations of orthophosphate and total
soluble inorganic nitrogen. Addition of EDTA before assay can help determine
nutrient enrichment problems in river systems receiving heavy metal wastes.
(Abstract-Buchanan-Davidson-Wisconsin WRSIC).
0078
Miller, W. E. , J. C. Greene, and T. Shiroyama. 1976. Use of algal assays to
define trace-element limitation and heavy metal toxicity, p. 317-325. In
Proc. Symp. Terr. Aquat. Ecol. Studies of N.W. , EWSC Press, Cheney, Wash.
71
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heavy metals, trace elements, algal assay, bottle test, zinc, copper, cadmium,
inhibition, algae, Selenastrum capricornutum.
Recent attention has focused on the biological response to conditions of
trace-element deficiency and heavy metal stress. This approach is necessi-
tated, in part, by the fact that metals are bonded by ligands and particles in
varying degrees. Therefore, absolute metal concentrations contained in the
wastewater effluent or receiving water do not necessarily reflect the degree
to which they affect biological organisms. The Algal Assay Procedure Bottle
Test has been used to define the toxicity of zinc, copper and cadmium under
controlled laboratory conditions. Those results indicate that the sensitivity
of Selenastrum capricornutum to these metals is inversely proportional to the
ionic strength of the test substrate. Other factors affecting toxicity, such
as increasing metal concentration, contact time and environmental test charac-
teristics, were also evaluated. Inhibition by zinc, 0-100 percent, was linear
with increasing concentration while increasing the concentration of both
copper and cadmium resulted in linear inhibition up to 20 and 40 percent,
respectively. Length of contact time, up to 21 days, reversed inhibition of
growth of the test organism when exposed to sublethal concentrations of cop-
per, cadmium, and zinc. (Abstract-Author).
0079
Miller, W. E. , J. C. Greene, and T. Shiroyama. 1978. The Selenastrum capri-
cornutum Printz Algal Assay Bottle Test: Experimental design, applica-
tion, and data interpretation protocol. U.S. Environmental Protection
Agency, Corvallis, Oregon. EPA-600/9-78-018. 125 p.
algal assay, bottle test, wastewater, nutrients, inhibition, heavy metals,
lake waters, rivers, data interpretation, experimental design, trace elements,
pretreatment, toxicity, algae, Selenastrum capricornutum Printz.
This document is the product of intensive research to improve and expand
the understanding of results obtained from the Algal Assay Procedure Bottle
Test (USEPA, 1971) to enable investigators to define the stimulatory and/or
inhibitory interaction(s) of municipal, industrial and agricultural wastes
upon algal productivity in natural waters.
This research was designed to determine:
(1) The impact of nutrients and/or changes in their loading upon algal
productivity;
72
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(2) Whether the growth response of Selenastrum capricornutum reflects
the response of indigenous species;
(3) The feasibility of the assay test protocol to evaluate heavy metals;
(4) The capability of the assay to define the effect(s) of complex
wastes; and
(5) If the assay information can be applied to define and assist in the
management of real-world situations. (Preface-Author).
0080
Miller, W. E. , T. E. Maloney, and J. C. Greene. 1974. Algal productivity in
49 lake waters as determined by algal assays. Water Res. 8(9): 667-679.
lake waters, algal assay, phosphorus, nitrogen, limiting nutrients, algae,
Selenastrum capricornutum.
Algal assays were conducted on waters from 49 American lakes to determine
whether phosphorus, nitrogen, or some other nutrient was the limiting factor
upon algal productivity.
A comparison of the known trophic state of 23 American lakes and their
bioassay response is presented. Phosphorus limited algal growth in 35 of the
49 lakes assayed. Nitrogen was limiting in eight lakes and constituents other
than nitrogen and phosphorus were limiting to six lakes. The results of this
study indicated phosphorus limitation decreased as the productivity of the
lake waters increased. (Abstract-Author).
0080-a
Miller, W. E. , J. C. Greene, E. A. Merwin, and T. Shiroyama. 1978. Algal
bioassay techniques for pollution evaluation, p. 9-16. Ijn Toxic mater-
ials in the aquatic environment. Seminar Oregon St. Univ., Water Res.
Res. Inst., Spring 1978, Corvallis, SEMIN WR 024-78.
toxicity, algal assay, bottle test, eutrophication, water quality, heavy
metals, limiting nutrients, nitrogen, phosphorus, zinc, wastewater, stimula-
tion, inhibition, lake waters, impoundment, algae, Selenastrum capricornutum.
The paper is a discussion of the utilization of the Algal Assay Bottle
Test procedure in studying nutrient limitation, heavy metal toxicity and in
evaluating complex wastes. The test organism is the alga Selenastrum capri-
cornutum. The basic techniques involved in the procedure are included in the
73
-------
discussion of the development and application of the AA:BT procedure. (Ab-
stract- Lei schman-EPA).
0081
Miller, W. E. , J. C. Greene, T. Shiroyama, and E. Merwin. 1975. The use of
algal assays to determine effects of waste discharge in the Spokane River
system, p. 113-131. Ij} Proc. Biostim. Nutr. Assess. Workshop, 16-17
October 1973. U.S. Environmental Protection Agency, Con/all is, Oregon.
EPA-660/3-75-034.
wastewater, heavy metals, zinc, toxicity, inhibition, limiting nutrients,
nitrogen, phosphorus, bottle test, algal assay, algae, Selenastrum capri-
cornutum.
This paper is a report on the assessment of waters impacted by industrial
and domestic waste effluents and their effect on algal growth. The area of
investigation was the Spokane River system, Idaho and Washington. Algal
assays were performed on samples collected at seven sites along the Spokane
River system. The assays were conducted to determine the nutrient status of
the waters (nitrogen and phosphorus concentrations); the critical nutrient
responsible for algal growth in the river system; and, the level of zinc that
would prevent algal growth (algistatic level). The authors found that in
sampled areas of the river system where heavy metals, particularly zinc, were
not dominant, additions of nitrogen and phosphorus stimulated algal growth;
however, the process reversed in waters below the locations of smelter and
mining industries, growth decreasing proportionally to increase in zinc con-
centrations. The method used to remove zinc from the sample waters and the
addition of a chelator (EDTA) both resulted in increased algal growth with the
nutrient additions. The algal assay was found to be effective in assessing
the problem of nutrient enriched waters containing heavy metals. (Abstract-
Lei schman-EPA).
0082
Mitchell, D. 1973. Algal bioassays for estimating the effect of added mater-
ials upon planktonic algae in surface waters, p. 153-158. I_n E. Glass
[ed.J, Bioassay techniques and environmental chemistry, Ann Arbor Sci.,
Mich.
algal assay, bottle test, PAAP, MAAP, NTA, trisodium nitrilotriacetate, diver-
sity indices, algae, Selenastrum capricornutum, Anabaena flos-aquae, Micro-
cystis aeruginosa.
74
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The Provisional Algal Assay Procedure (PAAP) Bottle Test and the Micro-
cosm Algal Assay Procedure (MAAP) were compared. The PAAP was found to be an
excellent analytical tool for assessing the physiological effects of various
materials upon algae, and for determining the limiting nutrient in water
samples. However it lacks the capability, which the MAAP does have, of delin-
eating interactions between the ecosystem and the material being assessed, as
well as between the various algal species making up the natural community.
Examination of sodium nitrilotriacetate (NTA) showed no significant effects
even at 200 mg/liter (at least 1000 times the expected environmental levels)
in the PAAP. Likewise, NTA showed no effects in the MAAP at the lower levels,
but at 200 mg/liter a stimulation of algal growth and loss of diversity was
observed. (Abstract-Author).
0083
Monahan, T. J. 1976. Lead inhibition of chlorophycean microalgae. J. Phy-
col. 12(3):358-362.
lead, phosphate, pH, toxicity, algae, Scenedesmus sp. , Scenedesmus obtusius-
culus, Selenastrum capricornutum, Chlorella pyrenoidosa, Chlorella ellipsoidea
Chlorella vulgaris, Ankistrodesmus sp. , algal assay.
The effects of Pb on the growth of 9 microalgae representing 4 chloro-
phycean genera were investigated in axenic laboratory cultures. Variations in
sensitivity to Pb existed among the algae tested with availability of phos-
phate often limiting growth. Under conditions in which phosphate was growth
limiting at pH 6.2 Chlorella pyrenoidosa Chick, C. ellipsoidea Gernick, C
vulgaris Beij. Scenedesmus sp. (IUCC 1591), S obtusiusculus Chodat and Sele-
nastrum capricornutum Printz first showed toxicity at 0.5 mg I-1 Pb. Two
planktonic Scenedesmus spp. and an Ankistrodesmus sp. were more tolerant,
developing toxicity at 1 mg I-1 Pb. The relationship of phosphate supply to
Pb toxicity was investigated with Selenastrum capricornutum and S. obtusius-
culus. At pH 8.0, Selenastrum developed toxicity at 3 mg I-1 Pb either with
or without phosphate. When phosphate supply was growth-1imiting, 6 times as
much Pb (i.e. 3 vs 0.5 mg I-1) was required to demonstrate toxicity with
Selenastrum at pH 8.0 as opposed to pH 6.2. S>. obtusiusculus was inhibited at
0.5 mgl-1 Pb in phosphate-limiting medium and developed toxicity at 1 mg • I-1
in media containing phosphate. Control of colonial morphology with S. pbtus-
75
-------
iusculus was inhibited. Phosphate availability and pH seem to be important
factors determining Pb toxicity with some algae. (Summary-Author).
0084
Morton, S. D. , and T. H. Lee. 1974. Algal blooms: possible effects of iron.
Environ. Sci. Technol. 8(7):673-674.
iron, manganese, phosphorus, algae, Chlorella pyrenpidosa, Dictyosphaerium
pullchellum, Selenastrum capricornutum, Anabaena circinalis, Gloeotrichia
echinulata, Microcystis aeruginosa, algal assay.
Iron, in concentrations of 0.1-1.0 mg/1, causes a shift in the dominant
type of algae grown in batch cultures from relatively unobjectionable greens
to the scum-forming blue-greens, without causing a significant change in the
total algal biomass. Results were not dependent on the manganese concentra-
tion (0-0.40 mg/1) or the phosphorus concentration (0.07-7.0 mg/1). (Ab-
stract-Author).
0085
Murray, S. , J. Scherfig, and P. Dixon. 1971. Evaluation of algal assay
procedures: PAAP batch test. J. Water Pollut. Control Fed. 43(10):1991-
2003.
algal assay, trace elements, nutrients, light intensity, hydrogen, carbon
dioxide, eutrophication, bottle test, algae, Selenastrum capricornutum.
Preliminary investigations have demonstrated four important difficulties
involved in the Provisional Algal Assay Procedure (PAAP) Batch Test. The four
difficulties were: (1) the effects of medium preparation on the final compo-
sition of the medium; (2) pH changes during the test period in relation to
carbon dioxide availability; (3) the method of gas addition in the batch test
units; and (4) the effects of light intensity on algal growth. Because of
these limitations, certain modifications to the standardized procedures were
developed. A new culture media preparation which eliminates the precipitation
of essential trace elements was developed and provides a reliable reference
medium for algal assay work. The addition of C02-enriched air reduced the
growth-limiting effects of C02 and stabilizes the pH at 7.5 to 8.0. Aeration
was shown to be more effective than ventilation for C02 addition and gaseous
as a result of bicarbonate utilization. Light intensities of 500 ft-c reduced
76
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growth rates of Selenastrum capricornutum in bottle test experimentation
compared with 350 ft-c. (Abstract-Lowry-Texas WRSIC).
0085-a
Natarajan, K. V. 1960. Mineral nutrition of Selenastrum westii, p. 156-161.
In P. Kachroo [ed.J, Symposium of Algology, Indian Council Agric. Re-
search, New Delhi.
algal assay, algae, media, nutrients, nitrogen, phosphorus, N:P ratios, Kirch-
neriella subsolitaria, Selenastrum westii.
The present observations showed that in a purely inorganic medium of
suitable composition, Selenastrum westii showed rapid growth and gave high
final yields of cells. It was observed that while the alga required about
20.4 ppm N for its maximum growth, the phosphate concentration required was
only 0.9 ppm P. The nitrogen:phosphorus ratio of 22.6:1 found for this alga
is in close agreement with the values found for other algae (Harvey, 1940;
Miller and Fogg, 1957). Kirchneriel la subsolitaria, another member belonging
to the same family was found to require an N:P ratio of approximately 15:1
(Potash, 1958). The minimum concentrations of the essential major elements
for the production of maximum numbers of cells of this alga were 20.4 ppm N,
Ca and 2.5 ppm Fe. The growth of the alga in this solution was found to be
135, 27.36 of that in the basic solution. (Abstract-Biological Abstracts,
Inc.).
NOTE: The above article was not located, as there were errors in trans-
lation of the title of book and the source. The National Agricultural Li-
brary, Dictionary Catalog of N.A.L., Vol. 59, U.S.D.A., does list the book
under the editor's name.
0086
Nyholm, N. 1977. Kinetics of phosphate limited algal growth. Biotech.
Bioengr. 19:467-492.
growth kinetics, limiting nutrients, phosphate, continuous flow, phosphorus,
Michaelis-Menten equation, chlorophyll a, algae, Chlorella pyrenoidosa, Selen-
astrum capricornutum, algal assay.
The kinetics of phosphate limited growth of two green algae Chlorella
pyrenoidosa and Selenastrum capricornutum have been studied in chemostats.
77
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Several kinetic models which express the specific growth rate as a function of
the intracellular phosphorus content have been examined, and one of the models
was found to be significantly better than the other models. The principles of
this model were described in a recent paper by Nyholm.
The kinetics of phosphate uptake have been investigated by adding pulses
of phosphate to the chemostats. The uptake by phosphorus deficient cells
could be described by Michaelis-Menten kinetics for phosphate concentrations
below approximately 500 (jg P/liter. Further, with the assumption of a discon-
tinuous adjustment of the uptake rate at the onset of phosphorus deficiency, a
complete kinetic model for growth and phosphate removal is proposed.
The mean cell size and the contents of chlorophyll a and RNA per unit dry
weight have been measured for C. pyrenoidosa as a function of the dilution
rate. (Summary-Author).
0086-a
Olofsson, J. A., Jr., and F. E. Woodard. 1977. Effects of pH and inorganic
carbon concentrations upon competition between Anabaena flos-aquae and
Selenastrum capricornutum. Land and Water Res. Inst., Univ. Maine,
Orono, Completion report.
inorganic carbon, competition, inhibition, pH, photosynthetic rates, uptake,
eutrophication, algal blooms, algae, Anabaena flos-aquae, Selenastrum capri-
cornutum, bottle test, algal assay.
Underlying reasons for seasonal dominance of lacustrine phytoplankton by
the blue-green algae have not heretofore been precisely defined. Relative
competitiveness of a selected green-alga and blue-green alga was examined in
relation to culture pH and total inorganic carbon (CT) in an effort to uncover
a mechanism which might be influential in blue-green algal bloom initiation.
Short-term growth experiments were conducted employing Anabaena f los-aquae, a
bloom forming blue-green alga and Selenastrum capricornutum, a green alga. A
self-contained polarographic dissolved oxygen electrode system was employed to
measure net photosynthetic oxygen evolution and respiratory oxygen consumption
at 25°C.
Media containing total inorganic carbon levels of 0.107, 0.214, 0.429 and
1.072 mg I-1 were employed at pH levels of 7.0, 8.0, 9.0 and 10.0. Algal
cells of uniform culture history were harvested in mid- to late log phase
78
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growth. They were washed and resuspended in the test medium and then placed
in the dissolved oxygen probe cell. Dissolved oxygen changes during light and
dark periods permitted calculation of oxygen evolution rates per unit mass of
algal material in the test suspension. The chosen CT levels imposed carbon
limitation upon the algal suspensions as evidenced by increasing photosyn-
thesis as CT increased at given pH levels. All other nutrients were present
in excess.
A. flos-aquae, at a given level of CT, exhibited increasing photosynthe-
tic rate as pH increased and reached a maximum in the region of pH 9.5 to
10.0. On the other hand, at given CT, levels, S. capricornutum exhibited
decreasing photosynthetic rates as pH was increased. Photosynthetic rates of
A. flos-aquae exceeded those of S. capricornutum above pH 7.5, indicating
potential pH-related enhancement of blue-green photosynthesis.
Carbonic anhydrase was implicated in facilitating photosynthetic rates in
A. flos-aquae by using a specific inhibitor of carbonic anhydrase activity.
Outside the pH range of 8.35 to 9.0, when the inhibitor Diamox (5-acetamido-
1,3,4-thiadiazole-2-sulphomanide) was introduced, significantly reduced photo-
synthetic rates were observed in A. flos-aquae. It was concluded that A.
flos-aquae has a higher affinity for inorganic carbon (as evidenced by photo-
synthetic rates) than does ^. capricornutum due to carbonic anhydrase ac-
tivity. Based upon the observation that Diamox had no significant effect upon
photosynthetic oxygen evolution within the pH range of 8.35 and 9.0, it was
concluded that carbonic anhydrase is not important here. Also, A. flos-aquae
may preferentially utilize HC03-carbon. The maximum kinetic rate of HC03
availability via carbonate alkalinity equilibria was equal to the maximum rate
of carbon fixation estimated from oxygen evolution rates.
The overall conclusion drawn from this investigation is that periodic
dominance of the lacustrine phytoplankton by bloom forming blue-green algae,
as represented by A. flos-aquae, may be related to competitive uptake of in-
organic carbon facilitated by carbonic anhydrase activity. Facilitated carbon
transport in blue-green algae may provide additional insight into the manage-
ment of blue-green algal blooms in surface water resources. (Abstract-
Author).
79
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0087
Oswald, W. J. , and S. A. Gaonkar. 1969. Batch assays for determination of
algal growth potential, p. 23-38. In E. J. Middlebrooks, T. E. Maloney,
C. F. Powers, and L. M. Kaack [eds.], Proc. Eutroph. Biostim. Assessmt.
Workshop, 19-21 June 1969, Berkeley, Calif. SERL and NERP sponsored.
algal assay, bottle test, growth rates, kinetics, trace elements, toxicity,
eutrophication, AGP, algae, Selenastrum capricornutum, Anabaena flos-aquae,
Microcystis aeruginosa, Chlorella pyrenoidosa, Scenedesmus obliquus, Euglena
gracilis.
Successful use of algal cultures in studies of algal growth potentials,
trace mineral analyses, tastes and odors, and toxicity, gives reason to be-
lieve they may be valuable for assessment of eutrophication potential in the
"Bottle Test" version of the Provisional Algal Assay Procedure (PAAP), as
proposed by the National Eutrophication Research Program. While tests may
lack elegance, they are simple, flexible, versatile, reliable, and appli-
cable—criteria established as desirable for that program. Author feels that
certain characteristics of the test, as presently recommended, require alter-
ation. It is doubtful, for example, if requirements for vigorous shaking,
rigid temperature control, and accessory gases are necessary. In addition,
simplified incubation techniques and apparatus should be evaluated. Several
of the techniques proposed for assessment of growth in cultures may be unnec-
essarily complex or expensive, whereas techniques of fluorescence analysis and
volumetric analysis of growth—not now being considered—should be studied.
Parameters of growth in PAAP could be evaluated in terms of growth levels for
which they may be most useful; thus, a rational system might involve enumera-
tion of cells for algal growth potentials (in milligrams/liter) of 0.1-1.0;
fluorescence, for 10-100; and gravimetry, for 100-1000. (Abstract-Eichhorn-
Wisconsin WRSIC).
0088
Palmer, M. 1969. A composite rating of algae tolerating organic pollution
J. Phycol. 5(l):78-82.
pollution, organics, algae.
From information on pollution-tolerant algae compiled from reports from
165 authors, the genera and species most often referred to as significant fall
80
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into a relatively stable series. Diatoms, pigmented flagellates, green, and
blue-green algae are all well represented among the pollution-tolerant genera
and species. The top 8 genera are Euglena, Oscillatoria, Chlamydomonas,
Scenedesmus, Chlorella, Nitzschia, Navicula, and Stigeoclom'um, and the top 5
species, Euglena viridis, Nitzschia palea, Oscillatoria limosa, Scenedesmus
quadricauda, and Oscillatoria tenuis. In some genera, e.g., Euglena, a single
species is far more significant than all others as a pollution-tolerant form.
In other genera, e.g., Oscillatoria, only a slight difference distinguishes
the polluton tolerance of 2 or more species. Algal genus and species pollu-
tion indices are presented for use in rating water samples with high organic
pollution. (Summary-Author).
0089
Paoletti, C. , B. Pushparaj, G. Florenzano, P. Capella, and G. Lercker. 1976.
Unsaponifiable matter of green and blue-green algal lipids as a factor of
biochemical differentiation of their biomasses: I. Total unsaponifiable
and hydrocarbon fraction. Lipids 11 (4):258-265.
lipids, hydrocarbons, sterols, phytoplankton, Spirulina platensis, Spirulina
sp. , Calothrix sp. , Nostoc commune, Scenedesmus quadricauda, Chlorella sp. ,
Uronema gigas, Uronema terrestre, Selenastrum gracile, carotenoids, unsapon-
ifiables, algae, algal assay.
As part of a program to study the chemical composition of algal bio-
masses, the composition of the unsaponifiable matter of the lipids of ten
algal species (five Myxophyceae and five Chlorophyceae) was investigated. The
total unsaponifiable content, its general composition, and the components of
the hydrocarbon fraction are discussed in the present paper. The unsaponifi-
able content of green algae is constantly higher than that of the blue-green
ones, with the exception of Chlorella. In both algal classes, the major
components are hydrocarbons and sterols. Blue-green algae are richer in
hydrocarbons, whereas the green ones contain higher amounts of sterols. In
most of the species examined, at least 48 components are present in the hydro-
carbon fraction. Each algal species shows a characteristic gas liquid chroma-
tography pattern, but n-C17 is always one of the most abundant components.
Generally, the procaryotic blue-green algae show a simpler hydrocarbon compo-
sition than the eucaryotic green algae, which contain higher amounts of high
mol. wt. components. Unsaturated hydrocarbons are generally present in very
81
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limited quantities, with the exception of Spirulina sp. , and Chlorella sp. ,
which contain a C17 alkene. Green algae also contain appreciable amounts of a
C27 monoene and of squalene. (Abstract-Author).
0090
Parker, M. 1977. The use of algal bioassays to predict the short and long
term changes in algal standing crop which result from altered phosphorus
and nitrogen loadings. Water Res. 11 (8): 719-725.
algal assay, bottle test, lake waters, phosphorus, nitrogen, mathematical
model, algae, Selenastrum capricornutum, standing crop.
Bioassays were performed on filtered water from Jackson Lake, Wyoming,
using the Algal Assay Procedure-Bottle Test with Selenastrum capricornutum as
the assay organism. It is hypothesized that (i) short- and long-term addi-
tions of phosphorus (P) and nitrogen (N) to waters should be evaluated sepa-
rately, and (ii) only bioassay treatments of lake water plus phosphorus plus
nitrogen (LW + P + N) should be used to predict the effect of long-term phos-
phorus additions on algal standing crop. Equations are developed directly
from the corresponding bioassay responses to predict short-term increases in
the lake's algal standing crop. Accepting the hypothesis, an equation de-
scribing the response of S. capricornutum to LW + P + N treatments is derived
to predict the long-term effects of phosphorus additions on algal standing
crop. This equation is then combined with another derived to describe the
steady state concentration of phosphorus in lake water. The resultant equa-
tion predicts the percent change in algal standing crop as a function of any
change in the lake's phosphorus loading. In most cases, use of LW + P rather
than LW + P + N treatments to predict changes in algal standing crop will lead
to quite different interpretation of bioassay data and probably to different
management decisions. (Abstract-Author).
0091
Parr, M. P. , and R. V. Smith. 1976. The identification of phosphorus as a
growth-limiting nutrient in Lough Neagh, using bioassays. Water Res.
algal assay, lake waters, limiting nutrients, nitrogen, phosphorus, waste-
water, bottle test, algae, Selenastrum capricornutum, phytoplankton, Aphaniz-
omenon flos-aquae, Oscillatoria redekei.
82
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Algae were used in monthly bioassays of Lough Neagh water to evaluate
nutrient limitation. Blue-green algae were limited by chelated iron; but once
this limitation was overcome both nitrogen and phosphorus were necessary to
produce a large growth response. The green alga Selenastrum showed a low
requirement for chelated iron. Addition of phosphorus was necessary in all
bioassays for prolonged algal growth, confirming the strategy of removing
phosphorus from sewage effluents to reduce algal levels in the Lough. (Ab-
stract-Author).
0092
Parra, B. 0. 1973. Qualitative study of phytoplankton of Laguna Verde,
Concepcion (Chile), excluding diatoms. Gayana Bot. 24:1-26. In Spanish.
phytoplankton, algae, classification.
Thirteen phytoplankton samples collected in Laguna Verde, Concepcion,
Chile, were studied. 35 species, 2 forms and 2 varieties of Fungi, Cyano-
phyta, Chrysophyceae and Chlorophyta were found. Four genera, sixteen species
and two varieties are new records for the microflora of Chile. These taxa are
Chlorobotrys regularis, Pleodorina californica, Polyedriopsis spinulosa,
Selenastrum gracile, Chodatella longiseta, Chodatella quadriseta, Franceia
droescheri, Franceia oval is, Scenedesmus dimorphus, Scenedesmus el 1ipsoideus
Scenedesmus opoliensis, Scenedesmus spinosus, Arthrodesmus octocornis, Stau-
rastrum irregulare, Staurastrum iverseni i var. americanum, Staurastrum poly-
morphum, Staurastrum tetracerum var. biverruciferum. (Translated Abstract-
Author).
0093
Payne, A. G. 1973. Environmental testing of citrate bioassays for algal
stimulation. Proc. 16th Conf. Great Lakes Res. 16:100-115.
algal assay, stimulation, growth rates, sodium citrate, bottle test, waste-
water, lake waters, inhibition, eutrophication, algae, Selenastrum capri-
cornutum, Microcystis aeruginosa.
Possible effects of sodium citrate on eutrophication were investigated
throughout a growing season using the bioassay procedure of the Algal Assay
Procedure Bottle Test. Primary and secondary treated sewage, with and without
83
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citrate, was added to filtered waters from 17 locations across the U.S.,
including Lake Erie and Lake Michigan. The algae selected for study were
bacteria-associated Selenastrum capricornutum and Microcystis aer_ugjno^a;
growth was followed by cell counts.
Sewage generally stimulated algal growth to higher maximum standing crops
than the control. Citrate added to sewage normally had no additional effect
on algal maximum standing crops and no stimulation by addition of citrate
alone was detected in any water. Citrate added to sewage prevented die-off of
Microcystis in four waters and citrate added without sewage treatment and at
relatively high levels of 1 and 10 mg/1 caused some growth inhibition of
Selenastrum in seven waters. In 156 test situations, citrate added to sewage
caused an increase in growth rate in eight situations and a decrease in growth
rates in six situations. In most instances, the different growth rates caused
no differences in maximum standing crops. (Abstract Only-Author).
0094
Payne, A. G. 1975. Responses of the three test algae of the Algal Assay
Procedure Bottle Test. Water Res. 9(4):439-445.
algal assay, bottle test, light intensity, nutrients, phosphorus, wastewater,
limiting nutrients, algae, Selenastrum capricornutum, Microcystis aeruginosa,
Anabaena flos-aquae.
The Algal Assay Procedure (AAP) Bottle Test utilized three test organisms
which, under standardized culture conditions, give a range of responses to the
physical and nutritional states of test waters. The test algae of the AAP are
Selenastrum capricornutum, Microcystis aeruginosa and Anabaena flos-aquae.
The responses of these organisms to light intensities and to micro- and macro-
nutrients are described. Phosphate spikes as low as 1-2 ug P I-1 are shown to
give a measurable bioassay response in highly oligotrophic waters.
The AAP test algae are used to assess the effects of nutrient enrichment
with sewage in waters from a cross-section of U.S. lakes. Enrichment of
oligotrophic waters with primary or secondary sewage effluent results in
significant growth of the three algae, while sewage additions to eutrophic
waters cause little or no significant increase in algal growth rate or maximum
standing crop. The response of the AAP algae also indicate that nutrient
removal by tertiary treatment greatly reduces the stimulatory properties of
84
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sewage. In the presence of tertiary sewage, growth rate and maximum standing
crop of the three test organisms are near those of lake water controls.
The AAP test protocol is shown to be useful in the determination of
limiting nutrients and in the comparison of the algal growth potential of
natural waters to that of defined media. (Abstract-Author).
0095
Payne, A. G. 1976. Application of the algal assay procedure in biostimula-
tion and toxicity testing, p. 3-28. .In E. J. Middlebrooks, D. H. Falken-
borg, and T. E. Maloney [eds.], Biostimulation and nutrient assessment,
Ann Arbor Sci. , Mich.
algal assay, bottle test, lake waters, wastewater, eutrophication, phosphorus,
chlorophyll a, trace elements, chelation, heavy metals, toxicity, Microcystis
aeruginosa, Anabaena sp., Navicula seminulum, phytoplankton, inhibition,
algistatic, algicidal, algae, Selenastrum capricornutum, diatoms.
The algal assay procedure, used in conjunction with physical/chemical
measurements of natural waters has been successfully applied to the assessment
of the algal growth potential of those waters.
In testing nutrient conditions of natural waters, the AAP showed logical
differences among waters and by seasonal patterns. Samples from oligotrophic
waters, when filtered, inoculated and maintained under laboratory culture
conditions, supported low standing crops of algae throughout the year. Natu-
ral populations in these waters also remained low throughout the year- Eutro-
phic waters during the winter months were rich in nutrients and supported high
maximum standing crops of test algae in AAP tests. The following spring,
these lakes, as predicted, supported high natural population levels.
The AAP can provide useful information on the relative chelating capacity
of synthetic compounds, sewage effluent, and natural chelators in the aquatic
environment. The sensitivity of laboratory-cultured Microcystis aeruginosa to
low levels of free metals makes this a useful test organism for trace metal/
chelator studies. The level of Zn in AAP media is close to the toxicity level
for Microcystis and may need to be lowered for optimization of Microcystis
growth.
The application of the AAP has been expanded to include an assessment of
algal toxicity of test materials. A toxicity test is described which is both
operationally simple and environmentally meaningful. The Minimum Algistatic
85
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Concentration after 5 days represents that level of a test material which
causes no net change in the population of the test algae after chronic expo-
sure. Inclusion of a recovery period following this contact allows interpre-
tation of the MAC-5 day to be the level of the test material which causes
primary inhibitory effects at the cellular level but does not cause permanent
or irreversible damage to the test culture. (Summary-Author).
0096
Payne, A. G. , and R. H. Hall. 1978. Application of algal assays in the
environmental evaluation of new detergent materials. Mitt. Int. Ver.
Limnol. 21:507-520.
algal assay, bottle test, detergents, phosphate, wastewater, chelation, trace
elements, toxicity, algae, Selenastrum capricornutum, Microcystis aeruginosa,
Anabaena flos-aquae, stimulation.
Algal assays have been used extensively in the environmental safety
evaluations of detergent candidate materials. The Algal Assay Procedure
Bottle Test (AAP), developed in cooperation with the U.S. Environmental Pro-
tection Agency, has been successfully applied, with modifications, to studies
of stimulation, chelation, and toxicity effects.
Algal stimulation studies have been conducted on materials considered for
use as detergent phosphate substitutes. These materials have been tested at
realistic and exaggerated environmental concentrations, alone and in the
presence of sewage, in surface waters from across the U.S. which differed in
hardness and varied from infertile to highly fertile.
The AAP ha*s been used to develop information on the relative chelating
ability of detergent builder candidates in defined systems. A comparative
chelating capacity test has been developed by altering the trace metal/ che-
lator system of AAP medium. Results of this test demonstrate the ability of
specific chelators to protect against metal toxicity and give some measure of
effects of detergent builders on the aquatic environment.
The AAP has been further modified to provide an assessment of algal
toxicity of detergent surfactants. A dose/response relationship can be ob-
tained which quantifies the effects of concentrations of test materials on net
cell production. The test is both operationally simple and environmentally
meaningful. (Abstract-Author).
86
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0097
Payne, A. G., and R. H. Hall. 1978. A method for measuring algal toxicity
and its application to the safety assessment of new chemicals. Proc.
Sec. Symp. Aquat. Tox., 31 October and 1 November 1977, Cleveland, Ohio.
ASTM publ. In press.
algal assay, bottle test, toxicity, algistatic, algicidal, trace elements,
inhibition, algae, Selenastrum capricornutum, Microcystis aeruginosa, Navicula
seminulurn, Dunaliella tertiolecta, diatoms.
An algal toxicity test has been developed specifically for use in the
environmental safety assessment of new chemicals. The test is based on the
Algal Assay Procedure Bottle Test, but has been modified in order to make the
procedure useful for toxicity assessment. A five-day exposure of freshwater
or marine unialgal cultures to a range of test material concentrations is
followed by a nine-day recovery period in the absence of the test material.
Cell counts or i_n vivo fluorescence measurements made during the study allow
quantification of toxic responses ranging from reduced growth to algistatic to
algicidal. The primary toxicity effect sought in this method is the algista-
tic response, which is both easily determined and environmentally meaningful.
The algistatic concentration of a test material, as derived by this method, is
the concentration that causes no net change in cell number after chronic
exposure but permits regrowth when the cells are resuspended in fresh media
without the test material. The environmental significance of this type of
measurement relative to other measurements of algal toxicity is discussed.
(Abstract-Author).
0098
Plumb, R. H. , Jr. 1975. A bioassay dilution technique to assess the signifi-
cance of dredged material disposal, p. 335-346. Ir\ E. J. Middlebrooks,
D. H. Falkenborg, and T. E. Maloney [eds.], Biostimulation and nutrient
assessment, Ann Arbor Sci. , Mich.
nutrients, nitrogen, phosphorus, sediments, interstitial waters, algal assay,
bottle test, dredging, algae, Selenastrum capricornutum, Dunaliella tertio-
lecta.
A potential problem at open-water dredged material disposal sites is that
nutrients (nitrogen and phophorus) associated with dredged sediment and inter-
stitial water may stimulate algal growth. Conventional bioassays are not
87
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applicable, since they only provide information on nutrient bioavailability in
dredged material, not the significance of discharge, as conventional bioassays
are run for long periods compared to the duration of dredged material pertur-
bation. A bioassay is proposed; based on nonlinear diffusion data observed in
the field, approximating changing concentration-time relationships at disposal
sites. Elutriates were prepared by shaking sediment with site water, then
allowing it to settle. After centrifugation and filtration, samples were
inoculated with Dunaliella tertiolecta or Selenastrum capricornutum. At times
selected by analysis of dye diffusion data, cultures were diluted to approxi-
mate what may happen at the disposal sites. Results with Dunaliella suggested
that open-water disposal of Mobile Harbor (Alabama) sediments would have no
significant effect on algal population; concentrated elutriates were toxic to
Dunaliella. It is concluded that inhibitory and stimulatory additions would
not significantly affect algae when dilution rates at open-water sites are
considered. The method could provide information to help determine the appro-
priate method of dredged material disposal. (Abstract-Buchanan-Davidson-
Wisconsin WRSIC).
0098-a
Plumb, R. H. , Jr., and G. F. Lee. 1974. Phosphate, algae, and taconite
tailings in the western arm of Lake Superior. Proc. 17th Conf. Great
Lakes Res. 17:823-836.
phosphate, tailings, taconite, lake waters, algal assay, stimulation, toxici-
ty, eutrophication, bottle test, algae, Selenastrum capricornutum.
The purpose of this investigation was to evaluate the significance and
availability of phosphate associated with taconite tailings that are presently
being discharged into the Minnesota waters of Lake Superior. Laboratory
leaching studies showed that phosphate would not be leached from tailings in
measurable quantities during a three year period and that phosphate from this
source is insignificant compared to other sources in the study area. In
addition, laboratory bioassays with Selenastrum and Lake Superior algae as
well as i_n situ bioassays demonstrated tailings to be non-stimulatory and
non-toxic. Since an observation of the primary productivity distribution in
the western arm of Lake Superior showed no apparent effect in the discharge
area and agreed with the measured and calculated phosphate flux distribution,
88
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it was concluded that the discharge of taconite tailings would not contribute
to the eutrophication of the lake. (Abstract Only-Author).
0098-b
Polesco-Ionasesco, L. 1974. Biomass produced by green algae Selenastrum
bibraianum and Chlorella vulgaris: 157 cultivated in laboratory and out
of doors. Lucr. Gradinii Bot. Bureau. 1974:183-190.
algae, Selenastrum bibraianum, Chlorella vulgaris, algal assay.
NOTE: No other information is available for the above article. A li-
brary search conducted by Library of Congress was not sucessful. Errors in
the bibliographic citation made retrieval of the article impossible.
0098-c
Polescu, L. 1974. Some physiological processes in the alga Selenastrum
bibraianum. Analele Universitatii Bucuresti, Seria Stiintele Naturii.
23:63-68.
algae, Selenastrum bibraianum, algal assay.
NOTE: No other information is available for above article. A library
search conducted through Library of Congress was not sucessful. Errors in the
bibliographic citation made retrieval of the article impossible.
0099
Porcella, D. B. 1969. Continuous flow chemostat assays, p. 7-22. In E. J.
Middlebrooks, T. E. Maloney, C. F. Powers and L. M. Kaack [eds.], Proc.
Eutroph. Biostim. Assessmt. Workshop, 19-21 June 1969, Berkeley, Calif.
SERL and NERP sponsored.
continuous flow, algal assay, phosphorus, kinetics, algae, Selenastrum capri-
cornutum, phytoplankton, Chlorella sp. , Anabaena sp. , wastewater, nitrogen,
lake waters.
Because their dynamic properties more nearly approximate ecological
conditions than did batch cultures, continuous cultures operating in chemostat
mode (constant flow-rate) have been utilized in a variety of studies of algal
growth, including assessment of biostimulatory responses. Requirements for
steady state continuous flow systems and characteristics of ideal chemostats
with illustrations of idealized curves from an experiment concerning relative
89
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change in algal cell material and limiting phosphorus concentrations are
included. In transient phases of chemostat operation, phosphorus concentra-
tions are less than those calculated theoretically, probably due to "luxury"
consumption by algae. Considerations of mass balances and kinetics of algal
growth, useful in derivation of continuous cultures theory, need be applied to
natural ecosystems to elicit useful information on productivity, nutrient
cycling, and successional relationships, based on chemostat experiments de-
scribed, utilizing both Selenastrum gracile and natural algal populations as
"seed", are areas requiring further study, including the following problems:
multiple limiting factors; use of natural "speed" populations; excessive
washout and extinction of cultures in bioassay of oligotrophic waters; devel-
opment of synthetic media; and profiles of biomass analysis and nutrient
concentration during transient growth in order to establish appropriate kin-
etic parameters. (Abstract-Eichhorn-Wisconsin WRSIC).
0100
Porcella, D. B. , P. A. Cowan, and E. J. Middlebrooks. 1973. Biological
response to detergent and nondetergent phosphorus in sewage: Part I.
Water and Sewage Works 120(11):50-67.
detergents, growth rates, limiting nutrients, phosphorus, wastewater, reser-
voirs, algal assay, algae, Anabaena flos-aquae, Selenastrum capricornutum.
Sewage effluent from a suburban community of the City of Logan, Utah, was
collected during times when detergents were and were not used to ascertain
whether the elimination of phosphate detergents would affect algal growth.
Algal bioassays were conducted with Selenastrum capricornutum and Anabaena
flos-aquae using nutrient spikes to determine which nutrients were limiting.
Bioassays were also performed using secondary treated, detergent-free sewage
samples spiked with detergent to observe whether any increase in biostimula-
tion resulted. The nutrient spikes were NH4C1 , KH2P04, Fe, and trace ele-
ments, and NAAM solution for control.
Chemical analysis for the sewage samples showed that restricted use of
detergent resulted in 57 percent decrease in phosphorus content; alum treat-
ment also reduced phosphorus content. However, calculated phosphorus concen-
trations in the bioassay flasks for different dilutions of the secondary and
tertiary effluents of the two sewage samples indicated the P content was high
90
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as a result of the high level of P in the reservoir water used for dilution.
It is concluded that if the P content of the water is naturally high, the
addition of more P will not affect algal growth. The results of the algal
bioassays are not included in this part of the report. (Abstract-Little-
Battelle WRSIC).
0101
Porcella, D. B. , P. A. Cowan, and E. J. Middlebrooks. 1973. Biological
response to detergent and nondetergent phosphorus in sewage: Part II.
Water and Sewage Works 120(12):36-46.
algal assay, wastewater, toxicity- phosphorus, nutrients, algae, Selenastrum
capricornutum, An-abaena flos-aquae, reservoirs, growth rates, trace elements,
stimulation.
The responses are presented of Anabaena flos-aquae and Selenastrum capri-
cornutum to detergent and nondetergent phosphorus. Maximum growth and mean
growth rates are discussed with maximum growth being based upon the first and
last biomass measurements during the growth cycle (18 or 21 days). Mean
growth rate represents growth over a 7-day period. Correlation between par-
ticulate carbon (PC), optical density (OD) and suspended solids (SS) was
performed to establish the relations between the individual parameters.
Dilution studies were made to detect toxicity, indicated by unusual growth as
a function of dilution. Deionized and reservoir water were used to dilute
effluents from secondary and tertiary sewage treatment. In summary, the
results indicate the following conclusions: (1) no differences exist between
baseline and test samples; (2) response was greater in secondary than tertiary
effluents; and (3) iron and trace elements appeared to stimulate slightly
greater growth. Samples of various wash products were added to a solution of
two percent test secondary effluent in reservoir water. The results showed no
significant difference in maximum growth. It can be concluded that the addi-
tion of the wash products to treated sewage did not affect the bioassay re-
sponse. (Abstract-Holoman-Batelle WRSIC).
0102
Porcella, D. B., P. Grau, C. H. Huang, J. Radimsky, D. F. Toerien, and E. A.
Pearson. 1970. Provisional algal assay procedures: First annual report.
Sanitary Engr. Research Lab, SERL Report, Berkeley, Calif. SERL Report
No. 70-8. 180 p.
91
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eutrophication, algal assay, growth kinetics, continuous flow, bottle test,
nutrients, growth rates, Michaelis-Menten equation, rivers, lake waters,
wastewater, algae, Selenastrum capricornutum.
This report is a detailed coverage of the algal assay procedure with
emphasis on a comparison of batch and chemostat assays. The procedures are
given for both methods, including actual experiments conducted using the test
alga Selenastrum capricornutum and the batch (bottle test) assay and the
chemostat assay. An outlined conclusion is given by the authors. (Abstract-
Lei schman-EPA).
0102-a
Premazzi, G. , 0. Ravera, and A. Lepers. 1978. A modified turbidostatic
system for algal population studies. Mitt. Int. Ver. Limnol. 21:41-49.
algal assay, continuous flow, algae, Selenastrum minutum, growth rates.
A new type of turbidostats described with these important advantages: the
population density is controlled by a photodetector sensor with one threshold
and the difference between the photoelectrical relays is minimized. The
system, in addition to being able to compare the growth rate of algal popula-
tion cultivated on media with different macro- and/or micronutrients, may be
used to record quantitatively and continuously the effects of any pollutant on
the primary production of algal populations. The advantage to adopt this
turbidostat has been demonstrated by the results obtained from experiment on
the effects of low concentrations of Cd salt on a population of Selenastrum
minutum. (Summary-Author).
0103
Proctor and Gamble. 1968. Proctor and Gamble laboratory data: PAAP test.
Unpublished report.
algal assay, bottle test, wastewater, lake waters, nutrients, algae, Sele-
nastrum capricornutum.
Not available for reproduction as requested by author.
92
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0104
Rawlings, G. D. 1978. Source assessment: Textile plant wastewater toxics
study - Phase I. U.S. Environmental Protection Agency, Washington, D. C.
Technology Series EPA-600/2-78-004h. 153 p.
wastewater, pollution, toxicity, stimulation, algal assay, bottle test, nutri-
ents, inhibition, algae, Selenastrum capricornutum.
The purpose of this study was to provide chemical and toxicological
baseline data on wastewater samples collected from textile plants in the
United States. Raw waste and secondary effluent wastewater samples were
analyzed for 129 consent decree priority pollutants, effluent guidelines
criteria pollutants, and nutrients; Level I chemical analyses were also per-
formed. Secondary effluent samples from the 23 plants selected for study in
the EPA/ATMI BATEA Study (American Textile Manufacturers Institute/best avail-
able technology economically achievable) (Grant No. 804329) were submitted for
the following bioassays: mutagenicity, cytotoxicity, clonal assay, freshwater
ecology series (fathead minnows, Daphnia, and algae), marine ecology series
(sheepshead minnows, grass shrimp, and algae), 14-day rat acute toxicity, and
soil microcosm. Since this was a screening study, samples of the textile
plant intake water were not collected for chemical analysis.
Based .on the bioassay results, 10 of the 23 textile plants were found to
have secondary effluents sufficiently toxic to proceed to a second phase of
the study. In the second phase, samples will be collected from these 10
plants to determine the level of toxicity removal attained by selected terti-
ary treatment technologies. (Abstract-Author).
0104-a
Reynolds, J. H., E. J. Middlebrooks, and D. B. Porcella. 1973. A continuous
flow kinetic model to predict the effects of temperature on the toxicity
of oil refinery waste to algae. Utah Water Res. Lab., NTIS, PB-239699,
U. S. Dept. Commerce, Springfield, VA. , 42 p.
oil, refinery, temperature, algal assay, continuous flow, wastewater, mathe-
matical model, inhibition, toxicity, algae, kinetics, Selenastrum capricornu-
tum.
Many industries, such as pulp and paper mills, tanneries, and oil refin-
eries, discharge toxic wastes at extremely high temperatures. The temperature
93
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of an oil refinery waste may range from 22°C (71 °F) to 41 °C (106°F). These
high temperatures could have a significant effect on the toxicity of a given
waste. It is therefore necessary to develop relationships which will evaluate
and predict the effects of increased temperature on the toxicity of various
wastes.
Equations have been developed utilizing continuous flow stirred tank
reactor (chemostat) kinetics and enzyme inhibition kinetics to describe the
effects of temperature on toxicity to microorganisms. These equations were
tested by semi-continuous and continuous flow experiments with phenol and the
algae Selenastrum capricornutum.
The data obtained were fitted to competitive, uncompetitive, and non-
competitive inhibitor models. The results indicated that phenol exerts a
competitive inhibition effect on the growth of S. capricornutum.
Comparison of the competitive inhibition constants, Kj, for S. capri-
cornutum exposed to phenol at 20°C, 24°C, and 28°C, indicate that phenol
toxicity increases with an increase in temperature according to the Arrhenius
function. (Abstract-Author).
0104-b
Reynolds, J. H., E. J. Middlebrooks, D. B. Porcella, and W. J. Grenney. 1974.
A continuous flow kinetic model to predict the effects of temperature on
the toxicity of waste to algae. Utah Water Res. Lab., NTIS, PB-239194,
U.S. Dept. Commerce, Springfield, VA. , 112 p.
continuous flow, toxicity, temperature, oil, wastewater, inhibition, phenol,
growth rates, algal assay, uptake, kinetics, algae, Selenastrum capricornutum.
A continuous flow kinetic model to describe and predict the effects of
temperature on the toxicity of a specific oil refinery waste to the green
alga, Selenastrum capricornutum, is developed. The model is based on enzyme
inhibition kinetics and is developed using semi-continuous flow and continuous
flow algal cultures grown between 20°C (68°F) and 33°C (91°F). Phenol is
employed as the controlling inhibitor or toxicant. The model is applied to
continuous flow algal cultures exposed to an actual oil refinery waste.
In addition, the maximum specific growth rates, u, the half saturation
constants, Kg3 and the nutrient utilization constants, K. and KR, for two
luxury uptake functions are determined for the alga, Selenastrum capricornu-
94
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turn, growing in an ammonium-nitrogen limited environment between 20°C (68°F)
and 33°C (91°F).
Results indicate that phenol and the oil refinery waste studied exert
competitive inhibition of Selenastrum capricornutum, and that phenol is more
toxic at 24°C (75°F) than at either 20°C (68°F) or 28°C (81°F). In addition,
the maximum specific growth rate, p, has a maximum value between 24°C (75°F)
and 27°C (81°F). Also, the ammonium-nitrogen half saturation constant, K ,
does not vary with temperature between 20°C (68°F) and 33°C (91 °F). The
variation of the nutrient utilization constants, K. and KB, for the luxury
uptake functions is similar to the variation of the maximum specific growth
rate, p, for the temperature range studied. (Abstract-Author).
0105
Reynolds, J. H., E. J. Middlebrooks, D. B. Porcella, and W. J. Grenney. 1975.
Effects of temperature on growth constants of Selenastrum capricornutum.
J. Water Pollut. Control Fed. 47(10):2420-2436.
algae, kinetics, Selenastrum capricornutum, temperature, bottle test, contin-
uous flow, nitrogen, growth rates, uptake, algal assay.
The alga Selenastrum capricornutum Printz has been specified for use in
bioassays by the Environmental Protection Agency. Continuous-flow (chemostat)
experiments conducted at 20°, 24°, 28°, and 33°C indicated that S. capri-
cornutum exhibits luxury uptake of ammonium-nitrogen. The luxury uptake of
the organism may be described by either a first-order or a hyperbolic func-
tion. The nutrient utilization constants for both luxury uptake functions
have a maximum value of 3.16 and 2.46 days, respectively, at 24°C. The con-
stants decrease as the temperature is increased or decreased from 24°C. The
maximum specific growth rates of the organism at 20°, 24°, 27°, 28°, and 33°C
are, respectively, 1.365, 1.992, 1.412, 1.390, and 1.274 days, the maximum
occurring between 24° and 27°C. The value of the ammonium-nitrogen half-
saturation constant was constant between 20° and 33°C with an average value of
5.356 ug/1 NH4N. (Abstract-JWPCF).
0106
Reynolds, J. H. , E. J. Middlebrooks, D. B. Porcella, and W". J. Grenney. 1975.
Effects of temperature on oil refinery waste toxicity. J. Water Pollut.
Control Fed. 47(11):2674-2693.
95
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algae, inhibition, mathematical model, oil, phenol, refinery, temperature,
toxicity, Selenastrum capricornutum, wastewater, continuous flow, algal assay.
A continuous flow kinetic model was developed to describe and predict the
effects of temperature on the toxicity of a specific oil refinery waste to the
green alga, Selenastrum capricornutum. The model is based on Michaelis-
Menten enzyme inhibition kinetics with ammonium nitrogen-limited, continuous
flow algal cultures between 20° and 28°C. Phenol is the controlling inhibi-
tor. The model was applied to continuous flow algal cultures exposed to an
actual oil refinery waste. The phenol and oil refinery waste studied exerted
competitive inhibition of Selenastrum capricornutum. Phenol was more toxic at
24°C than at either 20° or 28°C. The particular oil refinery waste used was
approximately 10 times more toxic than pure phenol to Selenastrum capricornu-
tum. (Abstract-JWPCF).
0107
Reynolds, J. H., E. J. Middlebrooks, D. B. Porcella, and W. J. Grenney. 1976.
Comparison of semi-continuous flow bioassays, p. 241-266. Iji E. J.
Middlebrooks, D. H. Falkenborg, and T. E. Maloney [eds.], Biostimulation
and nutrient assessment, Ann Arbor Sci., Mich.
algal assay, continuous flow, growth rates, mathematical model, temperature,
phenol, toxicity, inhibition, algae, Selenastrum capricornutum.
In semi-continuous flow cultures the organism specific growth rate,
substrate concentration, and cell concentration vary, while in continuous flow
cultures there is a steady state where organism specific growth rate, sub-
strate concentration, and cell concentration are theoretically constant. If
the withdrawal period in semi-continuous flow cultures is small enough, con-
tinuous flow cultures will be approximated. Actually continuous flow cultures
may be considered as a special case of semi-continuous cultures where number
of withdrawals per unit time is infinity. The critical factor is the time be-
tween withdrawal of culture content and addition of fresh substrates. An
experiment was conducted to develop a mathematical model to predict tempera-
ture effects on phenol toxicity to Selenastrum capricornutum and to compare
semi-continuous and continuous flow cultures. Both methods showed that phenol
was more toxic at 24°C than at 20°C or 28°C but the level of toxicity indi-
cated by continuous flow competitive inhibition constants was greater than
96
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that indicated by the semi-continuous flow constants. Continuous flow data
showed that phenol was more toxic as the hydraulic residence time increased.
These differences may be due to variation in the organism specific growth rate
which is observed in semi-continuous cultures and may mask certain toxic
effects. (Abstract-Buchanan-Davidson-Wisconsin WRSIC).
0108
Rodhe, W. 1978. Algae in culture and nature. Mitt. Int. Ver. Limnol. 21:7-
20.
algal assay, cultures, media, limiting nutrients, nutrients, lake waters,
PAAP, Z-8, experimental design, algae, Selenastrum capricornutum.
The report is a general discussion of the use of algal assays; the devel-
opment, continuation and future outlook. The author discusses the development
of media and the two media used now in algal assays, Z-8 and PAAP; also the
test organism, Selenastrum capricornutum; nutrients; growth interactions;
criteria of the algal cultures; and practical and theoretical conclusions.
(Abstract-Leischman-EPA).
0108-a
Ryding, S. 0., A. Claesson, and A. Forsberg. 1978. Research on recovery of
polluted lakes. II. Biomass budgets using natural and potential (algal
assay) chlorophyll a. Verh. Int. Ver. Limnol. 20(2):833-839.
lake waters, chlorophyll a, algal assay, nitrogen, phosphorus, algae, Sele-
nastrum capricornutum, nutrients.
Biomass budgets in terms of total chlorophyll a (= natural + potential
chlorophyll a), are presented for four polluted Swedish lakes, where nutrient
input was reduced due to chemical treatment of wastewater and sewage diver-
sion. A fertilizing capacity-availability-biomass budget (F.A.B.)-diagram was
constructed to illustrate the mass balance of the nutrients available for
algal growth.
The contributions from the wastewater treatment plant ranged between
11--40 o/o, 11—53 o/o and 9—78 o/o of the total external loading for N, P
and total chlorophyll a, respectively. For one lake the monthly variation in
biomass transports are also illustrated. Comparisons are made between nutri-
ent and biomass budgets. (Summary-Author).
97
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0109
Sachdev, D. R. 1973. Effect of organic fractions from secondary effluent on
algal growth. Ph.D. thesis, Rensselaer Polytech. Inst., Troy, N.Y.
402 p.
wastewater, organics, fractionation, stimulation, inhibition, phosphorus,
nitrogen, algae, Selenastrum capricornutum, Anabaena flos-aquae, Microcystis
aeruginosa, carbon-14, bottle test, algal assay.
An evaluation regarding the effect of sewage effluents and their organic
fractions on three test algal species, Selenastrum capricornutum (a green
alga), Anabaena flos-aquae (a nitrogen fixing blue-green alga) and Microcystis
aeruginosa (a non-nitrogen fixing blue-green alga), was made. Effluents of
domestic wastes from water pollution control facilities (a trickling filter
plant in Lake George, N.Y., and a contact stabilization plant, Clifton Knolls,
in Elnora, N.Y) were selected for study. The sewage effluents were collected,
membrane filtered (0.45 p) and then concentrated by the freeze-drying tech-
nique in order to get a sufficient amount of organics in the concentrated
effluent.
Sephadex Gels G-10, G-15, and G-25 were employed for separation. Frac-
tionation of concentrated effluent in G-10 resulted in five well defined
organic fractions. The frontal peak of G-10 was chromatographed on G-15
yielding two organic fractions. The frontal peak of G-15 was fractionated on
G-25 and only one organic fraction was obtained. The apparent molecular
weight (AMW) was assigned to each fraction from the calibration curves devel-
oped for each Sephadex Gel Column. Soluble organics, in the concentrated
effluents, with AMW <700 formed 62.2% (Lake George) and 58.5% (Clifton Knolls)
and of AMW >5000 were 20.4% (Lake George) and 28.8% (Clifton Knolls).
Algal cultures were grown in New Algal Assay Medium under controlled
environments of temperature, light and shaking. Concentrated effluents and
their organic fractions were added to the cultures of the three algal test
species and their effect was followed by daily absorbance and 14C (cpm) mea-
surements. 14C experiments were conducted in two different ways.
Maximum specific growth rate (u ) and maximum standing crop were em-
max
ployed to evaluate the growth response of the test algal species. |j was
nicix
calculated by EPA (1971) method and regression analysis. Maximum standing
crop was assumed to have been achieved in a flask when daily increase in
98
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biomass was less than 5%. Results were considered significant if deviation
from the control was more than ±10%.
Based upon the results generated, it was concluded that in general both
concentrated effluents and organic fractions with AMW <700 indicated-signifi-
cant stimulatory effect to u and maximum standing crop. Organic fractions
niBX
of AMW <700 had varying effects on different algal species, some showed stimu-
lation, some depicted inhibition while others had no effect.
The stimulatory effect was due to the organic compounds in the fractions.
These organic compounds acted as growth substances and substrates (nutrients).
In some organic fractions they acted in both capacities.
It was also noted that removal of organics from concentrated effluents or
reduction of their concentration by dilution could arrest algal growth.
The dual nature of organic fractions was noticed. Some organic fractions
would act differently when alone than when in combination with others. Some
organic fractions were stimulatory to one alga and inhibited another.
Some of the organic fractions showed better stimulation than the concen-
trated effluent.
The addition of phosphorus and nitrogen did not significantly affect the
algal growth response and it was concluded that the effect, noted in this work
was due to organic compounds in the fractions. (Abstract-Dissertation Ab-
stracts Internatl.).
0110
Sachdev, D. R. , and N. L. Clesceri. 1978. Effects of organic fractions from
secondary effluent on Selenastrum capricornutum (Kutz). J. Water Pollut.
Control Fed. 50(7): 1810-1820.
wastewater, fractionation, organics, stimulation, algal assay, nitrogen,
phosphorus, inorganic carbon, inhibition, algae, Selenastrum capricornutum.
The secondary treated effluents from two domestic wastewater treatment
facilities were membrane filtered (0.45u), concentrated via freeze-drying
technique, and then fractionated. Sephadex gels G-10, G-15, and G-25 were
employed for fractionation. Effect of the concentrated effluents and their
various organic fractions on the maximum specific growth rate (u ) and
max
maximum standing crop of Selenastrum capricornutum was investigated. In
general the concentrated effluents and the organic fractions with apparent
99
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molecular weight >700 indicated stimulatory effect on |j and the maximum
nicix
standing crop. The stimulation of S. capricornutum was the result of the
organic compounds in these fractions. The supplemental nitrogen, phosphorus
or inorganic carbon added to the medium from the addition of the organic
fractions did not contribute to the stimulation of this alga. Neither the
concentrated effluents nor any of the organic fraction showed any inhibitory
effect on S. capricornutum. (Abstract-JWPCF).
0111
Saldick, J. , and J. F. Jadlocki. 1978. Solubilization of biological avail-
able phosphorus by autoclaving Selenastrum. Mitt. Int. Ver. Limnol.
21:50-55.
phosphorus, algal assay, bottle test, nutrients, pretreatment, inhibition,
algae, Selenastrum capricornutum, solubilization.
Autoclaving solubilized 70-85% of the phosphorus contained in Selenastrum
but only 15-45% appeared as soluble reactive phosphorus (SRP). The soluble
but non-reactive phosphate (TSP minus SRP) was generally not available for
cell replication in 21 days in the Algal Assay Procedure Bottle Test (US EPA,
1971), but there were some exceptions in which a portion of this fraction was
used. SRP was consistently available in the 21-day bottle test. Retardation
of growth rate due to removal of essential micronutrient(s) other than iron
was observed when the cell population at the time of autoclaving was suffi-
ciently high. Recently acquired surplus phosphorus was released as SRP in
greater yield (-^50%) by autoclaving than older surplus phosphorus.
The nutritional state of the algae present in bottle test water samples
influences the test results and must be considered in their interpretation.
(Summary-Author).
0112
Schelske, C. L. , E. D. Rothman, and M. S. Simmons. 1978. Comparison of
bioassay procedures for growth-limiting nutrients in the Laurentian Great
Lakes. Mitt. Int. Ver. Limnol. 21:65-80.
phosphorus, algal assay, nitrogen, lake waters, algae, Selenastrum capri-
cornutum Printz, phytoplankton, bottle test.
100
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A series of experiments conducted on the effects of phosphorus, nitrogen
and silica on the growth of algae showed that phytoplankton were phosphorus
limited in Lake Huron and Lake Michigan but not in Lake Erie. Three types of
experiments were used: a spiked test with Selenastrum capricornutum Printz, a
spiked test with natural phytoplankton assemblages and a fixed-level test with
natural phytoplankton assemblages in which depleted nutrients were replenished
periodically to maintain specified levels. A replicated factorial design was
used in all experiments. Levels of nutrients were ambient (existing levels),
plus three additions of phosphorus ranging from 5 to 15 |jg/liter, two addi-
tions of silica, 0.8 and 1.6 mg Si02/liter, and one addition of nitrate, 0.30
mg N/liter. Although other variables were used, changes in chlorophyll stand-
ing crops were utilized most frequently to determine whether treatments affec-
ted algal growth. Results indicate that experiments with natural populations
can be conducted either in the field or laboratory and that the naturally
occurring phytoplankton assemblages respond to treatments containing low
levels of nutrients and are not excluded by experimental conditions. Such
experiments with small nutrient additions therefore can be used to obtain
valuable information on the ecology of specific populations in natural phyto-
plankton assemblages. (Summary-Author).
0113
Scherfig, J. , P. S. Dixon, R. Appleman, and C. A. Justice. 1973. Effect of
phosphorus removal processes on algal growth. U.S. Environmental Protec-
tion Agency, Washington, D. C. EPA-660/3-73-015. 81 p.
algal assay, phosphorus, bottle test, continuous flow, wastewater, eutrophi-
cation, limiting nutrients, algae, Selenastrum capricornutum, phytoplankton.
Laboratory studies were conducted to improve algal assay techniques for
use in evaluation of sewage treatment processes.
Laboratory studies (batch and continuous cultures) were conducted at the
Santee California water reclamation plant to evaluate the effect of tertiary
waste treatment processes on the amount of algal growth in the treated efflu-
ent.
Laboratory studies were also conducted to determine the growth limiting
nutrients in each type of tertiary effluent.
101
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Field tests were conducted using special study ponds and the results of
the field tests were compared with the laboratory test results. The labora-
tory and field tests showed the same relative ranking for the treated efflu-
ents. (Abstract-Author).
0113-a
Scott, W. E. , M. T. Seaman, A. D. Connell, S. I. Kohlmeyer, and D. F. Toerien.
1977. The limnology of some South African impoundments. I. The physi-
co-chemical limnology of Hartbeespoort Dam. J. Limnol. Soc. South Afr.
3(2):43-58.
eutrophication, lake waters, impoundment, nutrients, nitrogen, phosphate,
algal assay, algae, Selenastrum capricornutum Printz, bottle test.
Hartbeespoort Dam is a eutrophic, warm monomictic lake with overturn
occurring in April. The lake is stratified for about six months of the year
and in late summer the stratification appears stable. Light penetration in
the dam depends both on the inflow of turbid water as well as on the magnitude
of the algal population. In summer the surface waters are often super-
saturated with oxygen, while the hypolimnion is anaerobic. The concentrations
of nitrogen compounds, phosphate, Na, K, Ca, Cl and S04 are higher in waters
entering the dam via the Crocodile River. Nitrogen compounds are the primary
growth-limiting nutrients for algal growth and phosphate is the secondary
growth-limiting nutrient. Phosphate concentrations in the water have in-
creased approximately a hundredfold since 1928 and N03-N concentrations about
threefold. The pH as well as the mineral content of the dam have also in-
creased over the last 50 years. (Summary-Author).
0114
Sherrard, J. H. , and J. L. Hoyle. 1977. A productivity study of the Roanoke
River above Niagara Dam in Virginia. Va. Water Res. Res. Center, Blacks-
burg. Bulletin No. 104, 65 p.
rivers, lake waters, eutrophication, algal assay, bottle test, nutrients,
nitrogen, phosphorus, limiting nutrients, growth rates, wastewater, algae,
Selenastrum capricornutum, Anabaena flos-aquae, water quality.
Deterioration of the water quality in the upper end of the Roanoke River
of Smith Mountain Lake due to eutrophication has caused much concern. The
Smith Mountain Lake impoundment was constructed in 1965 as a pump storage
102
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facility to generate electrical energy and has served as a major recreational
area for Southwestern Virginia.
Eight sampling stations were established along the length of the Roanoke
River from an agricultural area in Montgomery County through the urban area of
Roanoke-Salem to Niagara Dam above Smith Mountain Lake. On 12 different occa-
sions, approximately once a month, grab samples were taken at the sampling
stations. Samples were analyzed for general chemical parameters and for
nitrogen and phosphorus. The Algal Assay Procedure Bottle Test [Environmental
Protection Agency, 1971] was conducted with the samples, and in addition, with
samples in which phosphorus and/or nitrogen spikes were added to determine
potential productivity and to determine which element was limiting productiv-
ity.
Results of this investigation demonstrate that the algal assay bottle
test is useful in correlating phosphorus concentrations with algal growth
rate. Phosphorus was found to limit the algal growth in the Roanoke River
above the Roanoke wastewater treatment plant for Selenastrum capricornutum and
Anabaena flos-aquae, the two algal species used in this study. Maximum spe-
cific growth rates tended to increase with increasing phosphorus concentra-
tion. (Abstract-Author).
0115
Shiroyama, T. , W. E. Miller, and J. C. Greene. 1975. Effect of nitrogen and
phosphorus on the growth of Selenastrum capricornutum Printz, p. 132-142.
In Proc. Biostim. Nutr. Assessmt. Workshop, 16-17 October 1973, U.S. En-
vironmental Protection Agency, Corvallis, Oregon. EPA-660/3-75-034.
algal assay, bottle test, nutrients, nitrogen, phosphorus, limiting nutrients,
stimulation, eutrophication, algae, Selenastrum capricornutum Printz.
The report discusses the use of the algal assay bottle test to determine
effects of nitrogen and phosphorus on algal growth. The test species used was
Selenastrum capricornutum. Tests were conducted in a defined medium. Nitro-
gen and phosphorus were added as nutrient spikes. The authors found that: in
the analysis of growth rates, maximum uptake of the nutrients nitrogen and
phosphorus occurred in the initial 5 days of growth; the increased growth
rates of the alga, with phosphorus additions indicated assimilation of phos-
phorus more rapidly than nitrogen; there was a defined linear relationship
103
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between produced biomass and concentrations of the nutrients; when all essen-
tial nutrients were present, toxicants were absent, and nitrogen and phos-
phorus requirements were determined, maximum growth could be predicted within
±20% of the actual yield. (Abstract-Leischman-EPA).
one
Shiroyama, T., W. E. Miller, and J. C. Greene. 1976. Comparison of the algal
growth responses of Selenastrum capricornutum Printz and Anabaena f los-
aquae (Lyngb.) De Brebisson in waters collected from Shagawa Lake,
Minnesota, p. 127-148. In E. J. Middlebrooks, D. H. Falkenborg, and T.
E. Maloney [eds.], Biostimulation and nutrient assessment, Ann Arbor
Sci., Mich.
algal assay, bottle test, growth rates, nutrients, nitrogen, phosphorus,
wastewater, limiting nutrients, algae, Anabaena flos-aquae (Lyngb.) De
Brebisson, Selenastrum capricornutum Printz, lake waters.
The growth response of Anabaena flos-aquae was studied to determine its
reliability as a test alga and to compare its growth response to that of
Selenastrum capricornutum in water samples from Shagawa Lake, Minnesota.
Results showed that A. f los-aquae can be used as a test organism to define
growth potential and nutrient status of lake waters. For counting, sonication
was more effecient than blending to fragment A. flos-aquae, but it damaged
cells so cannot be used to prepare an inoculum. Water containing more than
0.010 mg/1 ortho-phosphate yielded 0.45 mg/1 dry weight A. flos-aquae per
0.001 mg/1 phosphorus. Filtered and autoclaved-fi Itered water samples were
primarily nitrogen-limited for ^. capricornutum support. The A. flos-aquae
growth response indicated constituents other than nitrogen and phosphorus
limited its growth in filtered samples; addition of phosphorus stimulated
growth to the maximum predicted values in autoclaved-fiItered samples.
Comparative analysis of algal growth responses suggested that more than one
test species should be used to determine the growth potential or nutritional
status of a test water. If the presence of a biologically produced toxin is
suspected, filtered and autoclaved-fiItered samples must be analyzed to deter-
mine the presence of a biologically produced growth-inhibitor. (Abstract-
Buchanan-Davidson-Wisconsin WRSIC).
104
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0117
Shoaf, W. T. , and B. W. Lium. 1976. An evaluation of methods for measuring
algal growth. J. Res. U.S. Geological Survey 4(4):497-504.
algal assay, growth rates, electronic particle counter, fluorescence, algae,
Chlorella vulgaris, Selenastrum capricornutum, Chlamydomonas reinhardti i,
Scenedesmus quadricauda, Anacystis nidulans.
Algal growth was expressed as milligrams dry weight per litre with use of
the electronic particle counter for determining cell number and cell volume.
A gravimetric constant in milligrams dry weight per cubic micrometer was
determined. This constant was applicable throughout the growth curve since
there was a linear relationship between total cell weight and total cell
volume. Cell volume must be considered in measuring growth because there is a
change in average cell volume in addition to an increase in cell numbers
throughout the growth curve. Chlorella vulgaris and Selenastrum capricornutum
exhibited similar growth curves when growth was expressed as cells per litre
or as milligrams dry weight per litre. Chlamydomonas reinhardtii appeared to
have a lower algal growth potential when measured in cells per litre but had
the same growth potential as C. vulgaris and ^. capricornutum when expressed
as milligrams dry weight per litre. The growth rate for Scenedesmus quadri-
cauda was less than the other three, regardless of how the results were ex-
pressed. The image analysis system was used to count different types of algae
in a mixed culture. C. reinhardtii, C. vulgaris and Anacystis m'dulans were
not grossly distorted by the sample preparation method and could be distin-
guished from one another by variations in morphology. The counts could be
separated by computing the ratio of area to length for each particle and by
allowing the computer to sort and count each of the three types. Typical
ratios ranged from 5 to 7 for C. reinhardtii, 2 to 4 C. vulgaris, and 0.2 to
1.3 for A. m'dulans. The number of types to be counted in a mixture was
limited only by the ability to apply shape factors without having an overlap
between each type. Both the image analysis and electronic particle counter
produced equal count and size measurements. Fluorescence was a sensitive
measure of growth on a laboratory prepared medium since there was a loga-
rithmic relationship between fluorescence and cells per millilitre for both C.
vulgaris and S. capricornutum. Fluorescence measurements were not satisfac-
tory for determining growth in samples of varying chemical composition (which
105
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occurs in natural samples) because the chlorophyll-to-cell ratio varies with
media composition. This was especially pronounced with C. vulgaris. (Ab-
stract-Author).
0118
Shoaf, W. T. , and B. W. Lium. 1976. Improved extraction of chlorophyll a and
b from algae using dimethyl sulfoxide. Limnol. Oceanogr. 21(6):926-928.
chlorophyll a, chlorophyll b, DMSO, dimethyl sulfoxide, algae, Selenastrum
capricornutum, Chlorella pyrenoidosa, Tetraedron bitridens, Scenedesmus
quadricauda, Oocystis marssonii, Ankistrodesmus braunii, Chlamydomonas rein-
hardtii, Anacytis nidulans, Anabaena flos-aquae, Fremyella diplosiphon,
Cyclotella sp. , Nitzschia sp. , algal assay, diatoms.
Dimethyl sulfoxide (DMSO) and 90% acetone extracted equal amounts of
chlorophyll from diatoms and blue-green algae, but DMSO was superior to 90%
acetone for all green algae tested giving 2-60 times more chlorophyll depend-
ing on the species. The absorbance spectra of pure chlorophyll a (and b) from
600 nm to 750 nm were identical whether dissolved in 90% acetone or a mixture
of DMSO and 90% acetone (1:1 v/v). Thus, several equations for estimating
chlorophyll concentration based on extinction in 90% acetone are applicable
with this solvent. (Abstract-Author).
0119
Skulberg, 0. M. 1964. Algal problems related to the eutrophication of Euro-
pean water supplies, and a bioassay method to assess fertilizing influ-
ences of pollution on inland waters, p. 262-299. Iji D. F. Jackson [ed.J,
Algae and man, Plenum Press, New York.
algal assay, eutrophication, pollution, wastewater, lake waters, rivers,
phytoplankton, algae, Selenastrum capricornutum, Ankistrodesmus sp.
The author lists among the problems caused by excessive algal growth,
filter-blocking, impartation of taste and odor to water supplies, discolora-
tion of water, provision of nutrients for undesirable heterotrophs in pipes,
and formation of sediments in lakes. To illustrate differences in trophic
status among lakes, he compares two lakes in Norway:Lake Maridalsvannet (3.9
square kilometers/maximum depth, 45 meters) which provides Oslo with drinking
water, is oligotrophic, and shows little annual variation in its low concen-
trations of dissolved nutrients. Lake Borrevannet (1.8 square km/max depth,
106
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16 meters) is eutrophic and shows important annual changes with respect to
chemistry and water quality. Author states that Lake Lugano, Northern Italy;
has eutrophied due to pollution enrichment, citing classic example of Lake
Zurich as evidence that lacustrine trophic changes are best recorded in the
sediments. Utilizing Selenastrum capricornutum, (or Ankistrodesmus) an organ-
ism requiring only a few days to attain peak growth, author devised a bioassay
to assess effects and concentrations of nutrients in natural waters. Such an
approach would appear to have its greatest value in comparative limnological
studies. (Abstract-Eichhorn-Wisconsin WRSIC).
0119-a
Skulberg, 0. M. 1966. Algal cultures as a means to assess the fertilizing
influence of pollution, p. 113-138. Jji 0. Jaag and H. Liebmann [co-chm],
Advances in water pollution research, Water Poll. Control Fed., Washing-
ton, D. C.
pollution, wastewater, rivers, lake waters, fjord, algal assay, nutrients,
cultures, media, algae, Selenastrum capricornutum, Skeletonema costatum,
Oscillatoria Vaucher, Chlorella oval is Butcher.
The paper is a discussion of experiments, and the results, conducted to
study eutrophication in relation to the pollution of Norwegian inland waters
and fjords. Algal assays were used to determine the nutrients available to
support algal productivity. The presentation includes discussion of the test
organisms, Selenastrum capricornutum, Skeletonema costatum, Oscil latoria sp.,
and Chlorella oval is Butcher; the culture media, Z-8; growth experiments in
the media and inland waters; and, the possibilities and limitations in using
algal assays. (Abstract-Leischman-EPA).
0120
Skulberg, 0. M. 1973. A comparative investigation of water from 15 European
lakes, p. 85-100. Jji Algal assays in water pollution research, Proc.
Nordic Symp., 25-26 October 1972, Oslo, Norway. NORDFORSK, Secretariat
of Environmental Sciences.
algal assay, lake waters, eutrophication, water quality, COD, phosphorus,
nitrogen, nutrients, classification, algae, Selenastrum capricornutum.
A report on investigations of the status of 15 European lakes. The algal
assay was used to assess the effects of enrichment of the lake waters as a
107
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result of human influence. Methods of sampling; preparation of samples;
analyses of nutrients; and, growth results for the test alga Selenastrum
capricornutum are given. A classification of the lakes in regards to the
trophic status was possible from the results obtained. (Abstract-Leischman-
EPA).
0121
Skulberg, 0. M. 1974. Observation and monitoring of water quality by use of
experimental biological methods. Verh. Int. Ver. Limnol. 19(3):2053-
2063.
algal assay, pollution, wastewater, eutrophication, nutrients, water quality,
algae, Selenastrum capricornutum.
The report is a discussion of the importance and the implications for
using experimental biological methods to assess the effects of pollution on
water systems. The author emphasizes use of observations of biological popu-
lations coupled with physical and chemical characteristics of the waters, as a
good practice in water quality management. Algal assays as a comparative
limnological method are discussed. Data and results from past studies con-
ducted on 15 European lakes are presented. (Abstract-Leischman-EPA).
0122
Skulberg, 0. M. 1976. Culture collection of algae. Norwegian Inst. Water
Res. (NIWR) Publ. Q91, Blindern, Norway, 9 p.
cultures, algae, Cyanophyceae, Chlorophyceae, Bacil lariophyceae, lake waters,
rivers, coastal waters, nutrients, trace elements, algal assay, Selenastrum
capricornutum Printz CHL 1, Selenastrum westii G. M. Smith CHL 17.
The report is a brief procedure of the collection of 57 isolate algal
species, most originating from Norwegian waters. A description of the media
used and the growth conditions are given. Included is a list of all the
species collected, isolated, the location and year of collection, and the
media used for each species. (Abstract-Leischman-EPA).
0123
Smock, L. A., D. L. Stoneburner, and J. R. Clark. 1976. The toxic effects of
trinitrotoluene (TNT) and its primary degradation products on two species
of algae and the fathead minnow. Water Res. 10(6):537-543.
108
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TNT, trinitrotoluene, algal assay, toxicity, bottle test, algae, Selenastrum
capricornutum, Microcystis aeruginosa, inhibition.
The effects of alpha trinitrotoluene (alpha TNT) and its primary degrad-
ation product (TNT cc) commonly referred to as "pink water", were determined
on members of two trophic levels. The growth responses of the algae Sele-
nastrum capricornutum and Microcystis aeruginosa were examined through static
bioassays. Death and behavioral responses of the fathead minnow (Pimephales
promelas) were determined using a proportional diluter. Alpha TNT and TNT cc
were both more toxic to the fathead minnow than to either species of alga.
Five and 15 mg I-1, inhibited S. capricornutum and M. aeruginosa growth,
respectively. TNT cc inhibited 55. capricornutum growth at concentrations
above 9 mg I-1; it was lethal to M. aeruginosa at 50 mg I-1, but stimulated
growth at lower concentrations. The 96-h LC50 values based on the death
response of the fathead minnow to alpha TNT and TNT cc were 2.58 and 1.60 mg
I-1, respectively. The 96-h EC50 values based on the behavioral responses
were 0.46 and 0.64 mg I-1, respectively. There was no response to concentra-
tions of 0.05 mg I-1 alpha TNT and 0.07 mg I-1 TNT cc. (Abstract-Author).
0124
Smolen, M. D. 1975. Prediction of algal growth potential from chemical
nutrient analysis. Va. J. Sci. 26(2):78.
reservoirs, watershed, nutrients, nitrogen, phosphorus, AGP, nitrate, phos-
phate, ammonia, algal assay, algae, Selenastrum, Anabaena.
Weekly water samples were obtained from the reservoir and major tributary
streams of the Occoquan watershed and analyzed for forms of nitrogen and
phosphorus. A portion of these samples were also subjected to AGP analysis
using Anabaena and Selenastrum. Distinctly different responses were noted for
the two test species on water samples from several locations. Correlation
analysis for AGP yield with chemical nutrient parameters indicates that ni-
trate and phosphate concentrations relate closely to the AGP yields observed.
Although ammonium ion concentration was observed to vary from extreme low to
extreme high levels, no significant positive correlation with AGP yield was
noted.
109
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A series of curves is presented which show observed AGP yield as a func-
tion of nitrate concentration at various phosphate levels. The results indi-
cate that nitrate concentration determines Selenastrum yield when phosphate
concentration is greater than 0.010 mg P/L. Anabaena, however, responds to
nitrate concentration only when total phosphate concentration is greater than
0.150 mg P/L. (Abstract Only-Author).
0125
Soltero, R. A., A. F. Gasperino, P. H. Williams, and S. R. Thomas. 1975.
Response of the Spokane River periphyton community to primary sewage
effluent and continued investigation of Long Lake. D.O.E. Project
WF-6-75-081. Completion Report. 117 p.
wastewater, rivers, nutrients, reservoirs, nitrogen, phosphorus, eutrophica-
tion, temperature, stratification, fractionation, organics, sediments, algal
assay, bottle test, algae, Selenastrum capricornutum, phytoplankton.
In part, the study examined the effects of primary sewage effluent from
Spokane, Washington on the diatom community in the Spokane River. Both nat-
ural and artificial substrates were used at each of those sampling stations,
one above and two below the outfall. In terms of individual species abun-
dance, mean diversity indices, floristic similarities between sampling sta-
tions and mean autotrophic indices, the diatom community at the last down-
stream station had begun to recover from the discharge.
Continued investigation of Long Lake and its tributaries has shown that a
major source of nutrients influent to the reservoir was the City of Spokane's
sewage effluent. Existing mean concentrations of inorganic nitrogen and
orthophosphate increased approximately twofold and sevenfold, respectively,
below the effluent. Of the total influent phosphate, 22 percent was retained
in the reservoir.
The reservoir was again determined to be highly eutrophic. Orthophos-
phate, nitrate nitrogen, silica and temperature were found to be important
variables regulating the size of the phytoplankton community. Long Lake's
seasonal phytoplankton periodicity, however, depended primarily upon strati-
fication, turbulence, light penetration and retention time.
Close interval fractionation of Long Lake sediment cores has provided a
reliable static record of the reservoir's trophic status since 1958. Organic
matter appeared to be stabilized prior to incorporation into the sediments.
110
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Nitrogen and phosphorus concentrations increased 1.4 times whereas diatom
production has increased sevenfold. A sediment accumulation rate of 24 mm
yr-1 was determined and does not appear to be excessive when compared to other
reservoirs. Vertical migration, leaching and internal recycling of phosphorus
from the sediments should have little effect, if any, on the reservoir's
trophic status once secondary sewage treatment with phosphorus removal is
initiated by the City of Spokane. (Abstract-Author).
0126
Soltero, R. A., D. M. Kruger, A. F. Gasperino, J. P. Griffin, S. R. Thomas,
and P. H. Williams. 1976. Continual investigation of eutrophication in
Long Lake, Washington. Verification data for the Long Lake Model.
D.O.E. Project WF-6-75-081. Completion Report. 64 p.
lake waters, eutrophication, reservoirs, wastewater, algal assay, bottle test,
nutrients, phosphorus, nitrogen, limiting nutrients, heavy metals, inhibition,
water quality, phytoplankton, algae, Selenastrum capricornutum Printz.
Long Lake, an impoundment of the Spokane River, was again determined to
be eutrophic. The major source of nutrients influent to the reservoir was
from the Spokane primary sewage treatment plant. During the study. Long Lake
had a mean exchange rate of 27 days, shortest of the four years studied
(Soltero, Gasperino and Graham, 1973, 1974; Soltero et al_. , 1975). Hypolim-
netic anoxia was not as extensive as previously determined (Soltero et al. ,
1973; 1974; 1975) due to lower reservoir retention times and the establishment
of an underflow of the Spokane River at stations 1 and 2 during August and
September.
Phytoplankton and zooplankton standing crops also approximated those of
previous years (Soltero et al_. , 1973; 1974; 1975) in species diversity, occur-
rence and abundance. Primary productivity was the lowest of the four years
studied (Soltero et al. , 1973; 1974; 1975). Reservoir algal assay data indi-
cated nitrogen to be the primary limiting nutrient (57 percent of the time)
and phosphorus the secondary limiting nutrient (30 percent of the time).
Nitrogen and phosphorus co-limitation occurred in 13 percent of the samples.
Heavy metal inhibition to Selenastrum capricornutum Printz existed in every
sample.
Ill
-------
Prior studies (Soltero et a_[. , 1973; 1974; 1975) have given a comprehen-
sive portrayal of Long Lake's dynamics and this base-line information has been
used by Battelle, Pacific Northwest Laboratories, in the calibration of a
water quality model. However, the simulated results must be verified with
"actual" field data in order to determine the sensitivity of the model's
predictions. The data collected during this study are representative of past
years and can be used for the verification of the Battelle water quality model
of Long Lake. (Abstract-Author).
0126-a
Sridharan, N. , and G. F. Lee. 1974. Algal nutrient limitation in Lake
Ontario and tributary waters. Water Res. 11(10):849-858.
nutrients, lake waters, nitrogen, phosphorus, stimulation, rivers, trace
elements, limiting nutrients, wastewater, toxicity, eutrophication, algal
assay, algae, Selenastrum capricornutum.
In nutrient-spiking studies of Lake Ontario and its tributary rivers of
Niagara, Black, Genesee, and Oswego waters both nitrogen and phosphorus were
required for stimulation of cultured and natural algae growth. Samples of the
Genesee and Oswego Rivers generally showed nitrogen stimulation. Also, many
of the samples from these rivers demonstrated stimulation due to the addition
of micronutrients. However, care must be exercised in interpreting micro-
nutrient stimulation as the micronutrient solution contained EDTA, a strong
complexing agent, which can eliminate toxic elements in the water. Essen-
tially all lake water samples showed phosphorus limitation. It is possible
that phosphorus removal at domestic wastewater treatment plants located in
these areas could be sufficient to cause phosphorus to be limiting. It is
also possible that advanced waste treatment could reduce the concentration of
the apparent toxicants in these rivers which would stimulate algae growth in
the rivers and nearshore lake waters due to excess of phosphorus present.
Even though the potential impact of phosphorus removal cannot be predicted for
these rivers and nearshore Lake Ontario waters, it is clear that such a prac-
tice will be of some benefit in reducing and possibly reversing the excessive
eutrophication trend. (Abstract-Auen-Wisconsin WRSIC).
112
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0126-b
Sridharan, N., and G. F. Lee. 1977. Algal nutrient availability and limita-
tion in Lake Ontario during IFYGL, Part III. Algal nutrient limitation
in Lake Ontario during IFYGL. U.S. Environmental Protection Agency,
Duluth, Minn., Ecol. Res. Series. EPA-600/3-77-046a. 46 p.
nitrogen, phosphorus, limiting nutrients, phytoplankton, lake waters, rivers,
algal assay, bottle test, algae, Selenastrum capricornutum.
This study was conducted on the potential significance of nitrogen,
phosphorus and micronutrients in limiting planktonic algal growth in Lake
Ontario and its major tributaries. Standard algal assay procedures were used.
Samples of the open waters of Lake Ontario and Niagara River waters collected
during the spring showed phosphorus limitation. By late summer these waters
showed both nitrogen and phosphorus limitation. Genesee and Oswego Rivers
showed, in general, nitrogen limitation. Samples of the Black River waters
showed both nitrogen and phosphorus limitation. (Abstract-Author).
0126-c
Starr, R. C. 1978. The culture collection of algae at the University of
Texas at Austin. J. Phycol. Suppl. 14:47-100.
algae, media, cultures, Selenastrum bibraianum Reinsch 324 (UTEX), Selenastrum
capricornutum Printz 1648 (UTEX), Selenastr.um gracile Reinsch B 325 (UTEX),
Selenastrum minutum (Nag.) Collins 326 (UTEX), Selenastrum 235 (UTEX), Selen-
astrum 747.
An updated, complete list of algae (a variety of strains and their class-
ifications) available to researchers. Sections on maintenance media and
special growth media included.
0126-d
Steeman Nielson, E. 1978. Principal aspects concerning the batch technique
in algal assays. Mitt. Int. Ver. Limnol. 21:81-87.
algal assay, growth rates, growth kinetics, algae, diatoms, Steeman Nielson
and v. Brand technique, Selenastrum capricornutum, Nitzschia palea.
The report is of experiments conducted using a special batch technique
(Steeman Nielson and v. Brand counting technique) to investigate influences of
low nutrient concentrations on dilute algal suspensions as might be found in
113
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nature. The author felt that the measurement of maximum growth rates in batch
cultures were not always the same as real growth rates. The experiments were
designed to run during the growth phase where growth rates were constant. The
two algae used in the experiments were Selenastrum capricornutum and Nitzschia
palea. The test containers were grown under continuous light and alternating
light and dark periods. The author found there was little difference in the
growth rates of the algae under the continuous light or 12 hr light and 12 hr
dark periods (radiance given as 28 quanta x 1015 x cm-2 x sec-1). Growth
curves are given. (Abstract-Leischman-EPA).
0127
Steensland, H. 1973. Application of the dialysis technique in testing pos-
sible growth retarding effects on algae in purified wastewater, p. 47-49.
In Algal assays in water pollution research, Proc. Nordic Symp., 25-26
October 1972, Oslo, Norway. NORDFORSK, Secretariat of Environmental
Sciences.
dialysis, growth rates, wastewater, toxicity. pollution, water quality, Z-8,
limiting nutrients, continuous flow, algae, Scenedesmus quadricauda, Sele-
nastrum capricornutum, algal assay.
A brief preliminary report on the use of the dialysis technique to test
effects of wastewaters on algal growth. An outlined procedure is presented
for the design of the dialysis bags and culture baths. References given.
(Abstract-Lei schman-EPA).
0128
Steyn, D. J. , D. F. Toerien, and J. H. Visser. 1974. Continuous culture
algal bioassays. South Afr. J. Sci. 70(9):277-278.
eutrophication, wastewater, limiting nutrients, continuous flow, nitrogen,
algal assay, algae, Selenastrum capricornutum.
A brief report of studies conducted using continuous flow algal assays to
determine the limiting nutrients of four South African impoundments. Nitrogen
was found to be the limiting nutrient in all four impoundments. Results and
discussion given. (Abstract-Leischman-EPA).
114
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0128-a
Steyn, D. J. , W. E. Scott, D. F. Toerien, and J. H. Visser. 1975. The eutro-
phication levels of some South African impoundments. I. Rietvlei Dam.
Water S.A. 1(2):45-52.
algal assay, impoundment, eutrophication, wastewater, nitrogen, phosphorus,
nutrients, bottle test, algae, Selenastrum capricornutum, Anabaena flos-aquae.
Algal bioassay results show Rietvlei Dam to be in a highly eutrophied
state. The growth of non-nitrogen-fixing algae is limited by nitrogen, and
that of nitrogen-fixing algae by phosphorus. It is predicted that increased
discharge of sewage works effluent will increase the eutrophication status of
the dam, but an inflow with lower nutrient concentrations will reduce it. The
results suggest that the control of phosphorus rather than nitrogen inputs
must be considered for eutrophication control, but a more complete understand-
ing of the chemical, physical and biological characteristics of the impound-
ment is required before a decision can be made as to the best methods of
eutrophication control. (Abstract-Author).
0128-b
Steyn, D. J. , D. F. Toerien, and J. H. Visser. 1975. Eutrophication levels
of some South African impoundments. II. Hartbeespoort Dam. Water S.A.
algal assay, bottle test, AGP, nitrogen, phosphorus, nutrients, eutrophica-
tion, impoundment, algae, Selenastrum capricornutum Printz.
Algal bioassay indicated that Hartbeespoort Dam is highly eutrophied
because algal growth potentials (AGP) of more than 100 mg I-1 and batch cul-
ture algal growth rates of 2.1 d-1 were recorded. Nitrogen was the primary
and phosphorus the secondary growth-limiting nutrient. Addition of secondary
sewage effluents linearly increased the AGP at rates of between 7.7 and 13.0
mg I-1 percent (v/v) effluents added. These increases could be attributed to
an increase in the nitrogen content of the water by the effluent. Addition of
sewage effluents did not change the identity of nitrogen as the primary
growth-limiting nutrient. Dilution with water of a low nutrient content
linearly decreased the AGP. Although nitrogen at present determines the
magnitude of algal blooms in the dam, phosphorus control is advocated for the
control of eutrophication problems in the dam. (Abstract-Author).
115
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0128-c
Steyn, D. J. , D. F. Toerien, and J. H. Visser. 1976. Eutrophication levels
of some South African impoundments. III. Roodeplaat Dam. Water S.A.
-
algal assay, AGP, limiting nutrients, phosphorus, wastewater, nitrogen, algae,
Selenastrum capricornutum, impoundment.
Algal bioassays indicated that the waters of Roodeplaat Dam are severely
eutrophied since algal growth potentials (AGP) of up to 100 mg/1 and batch
culture algal growth rates of up to 2.2 d-1 were registered with Selenastrum
capricornutum as test alga. Phosphorus was usually the primary growth"
limiting nutrient in the samples except at the end of winter when nitrogen was
primary growth limiting. The secondary limiting nutrient was either nitrogen
or phosphorus, depending on the identity of the primary growth-limiting nutri-
ent. Addition of secondary effluent linearly increased the AGP of Roodeplaat
Dam water by 7.2 to 12.4 mg/1 for each one per cent (v/v) of secondary efflu-
ent added. Addition of secondary treated sewage effluent resulted in a shift
to nitrogen as the primary growth- limiting nutrient. Dilution with a low
nutrient water linearly decreased the AGP. The control of phosphorus inputs
into the impoundment could be used to reduce the eutrophication level. (Ab-
stract-Author).
0128-d
Steyn, D. J. , D. F. Toerien, and J. H. Visser. 1976. Eutrophication levels
of some South African impoundments. IV. Vaal Dam. Water S.A. 2(2): 53-
57.
eutrophication, impoundment, wastewater, nutrients, nitrogen, phosphorus, AGP,
algal assay, algae, Selenastrum capricornutum.
The Vaal Dam shows algal bioassay growth rates and algal growth poten-
tials (AGP) as high as 1 . 6 d-1 and 146 mg/1-1 respectively using Selenastrum
capricornutum as test alga. Addition of secondary treated sewage effluents to
Vaal Dam water increased the AGP by between 7.4 and 11.2 mg/1-1 for every one
percent (v/v) added. Nitrogen and phosphorus were the important algal grow-
limiting nutrients in the impoundment. Plant nutrients adsorbed onto clay
particles could be important in the eutrophication of the impoundment, indi-
cating the importance of soil conservation techniques in eutrophication corr
116
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trol. The significance of the impoundment and the paucity in knowledge of its
physical, chemical and biological characteristics suggest that such studies
should be undertaken as a matter of urgency. (Abstract-Author).
0129
Sturm, R. N., and A. G. Payne. 1973. Environmental testing of trisodium
nitrilotriacetate: Bioassays for aquatic safety and algal stimulation,
p. 403-424. _In E. Glass [ed.], Bioassay techniques and environmental
chemistry, Ann Arbor Sci., Mich.
toxicity, wastewater, phosphate, NTA, trisodium nitrilotriacetate, algal
assay, bottle test, lake waters, iron, phosphorus, nitrogen, algae, Selenas-
trum capricornutum, Microcystij aeruginosa, Anabaena flos-aquae, Navicula
seminulum, fish, diatoms.
While data for chronic toxicity studies have not yet been released,
information is available from an extensive acute toxicity program which in-
cludes several species as well as three degrees of water hardness. The rela-
tive non-toxicity of trisodium NTA, even in very soft water where its effects
would be the greatest, and the absence of evidence of accumulative toxicity
suggest NTA to be compatible with the goal of environmental improvements.
While this work is but a small part of the total NTA environmental safety test
program, it does produce evidence that 1) NTA exerts no acute toxic effect on
the fish or algae species tested at levels well above those anticipated in the
environment, 2) Bluegills and fathead minnows exposed to high but sublethal
concentrations for a period of 28 days do not exhibit abnormal gill pathology,
a further indication of NTA's safety, and 3) NTA stimulatory effects on algae
would not be anticipated on the basis of nitrogen contribution and were not
found by application of the AAP Bottle Test to a variety of lake waters.
(Summary-Author).
0130
Sugiki, A. 1977. Algal growth potential of tertiary effluents. Japan Sewage
Works Agency. Dept. of Research and Development (unpublished).
eutrophication, wastewater, phosphorus, nitrogen, nutrients, algal assay, AGP,
water quality, stimulation, algae, Chlorella sp. , Selenastrum capricornutum,
Chlamydomonas sp., Stigeoclonium tenue, lake waters.
117
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Algal assays were used to determine the effects of the sewage effluents
and removal of phosphorus from the effluents on algal growth rates. The AGP
parameter was measured as the primary production rates within the water bod-
ies. Water samples from lakes and creeks of Japan were assessed. The test
algae used were Chlorella sp. and Selenastrum capricornutum. Results and
discussion are given for the AGP in natural waters and activated sludge -
treated effluents and effluents from other listed sewage treatment processes.
(Abstract-Lei schman-EPA).
0131
Thomas, N. A., K. Hartwell, and W. E. Miller. 1975. Great Lakes nutrient
assessment, p. 226-243. JTI Proc. Biostim. Nutr. Assess. Workshop, 16-17
October 1973, U.S. Environmental Protection Agency, Corvallis, Oregon.
EPA-660/3-75-034.
lake waters, nutrients, nitrogen, phosphorus, limiting nutrients, algal assay,
bottle test, algae, Selenastrum capricornutum, phytoplankton, Cladophora.
A series of studies were conducted to assess the nutrient status of the
Great Lakes. The first of the series was designed to determine the nutrient
requirements of the green alga Cladophora, a problem alga in the Great Lakes.
The studies were also to determine the availability of nutrients in the wa-
ters, especially those nutrients stimulating algal growth (nitrogen and phos-
phorus), and the effects of the addition of alum or iron salts to reduce
phosphorus concentrations. The final portion of the series was to find an
appropriate algal assay procedure for Great Lakes research, using the alga
Selenastrum capricornutum and natural phytoplankton populations. (Abstract-
Leischman-EPA).
0132
Thomas, R. E. , and R. L. Smith. 1975. Assessing treatment process efficiency
with the algal assay test, p. 244-248. I_n Proc. Biostim. Nutr. Assess.
Workshop, 16-17 October 1973, U.S. Environmental Protection Agency,
Corvallis, Oregon. EPA-660/3-75-034.
wastewater, nutrients, nitrogen, phosphorus, eutrophication, lake waters,
algal assay, bottle test, algae, Selenastrum capricornutum.
118
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The algal assay test was included in the evaluation of wastewater treat-
ment processes for feedlot runoff and for municipal wastewater. The biomass
productivity was directly related to the total nitrogen and/or phosphorus
concentration in the product water from the treatment process. The results of
these exploratory tests suggest that the algal assay test has promise for
comparing the eutrophying potential of the effluents from a variety of waste-
water treatment processes. This capability to measure eutrophying potential
could be an important factor in determining cost effectiveness of treatment
processes in many localities. (Summary-Author).
0133
Tison, D. L. , and A. J. Lingg. 1977. Algal bacterial mutualism in a simu-
lated aquatic environment. Annu. Meet. Am. Soc. Microbiol. 77(1):237.
organics, carbon, carbon dioxide, mutualism, glucose, stimulation, algae,
Anacystis nidulans, Anabaena flos-aquae, Chlorella pyrenoidosa, Selenastrum
capricornutum, algal assay.
The effects of increasing levels of organic enrichment in unialgal-
Dacterial cultures of non-limiting nutrient levels were determined. The
closed system of an electrolytic respirometer was employed to follow the fate
of added organic carbon. Glucose additions of 14, 64, and 128 mg C I-1 were
made daily. Glucose additions contained 3 uCi D-glucose-UL-14C. After six to
ten days each system was assayed for carbon dioxide evolved to the atmosphere,
inorganic carbon in solution, dissolved organic carbon, bacterial biomass
carbon, and algal biomass carbon. Bacterial metabolism of added substrate and
the resulting production of carbon dioxide and/or cofactors stimulated the
growth of the cyanobacteria Anacystis m'dulans and Anabaena flos-aquae and the
green algae Chlorella pyrenoidosa and Selenastrum capricornutum. Autotrophic
oxygen production provided a major portion of the oxygen required by the
heterotrophic community. The association between the autotrophic cyanobac-
teria and green algae and the heterotrophic bacteria in this system was mu-
tual istic. (Abstract Only-Author).
0134
foerien, D. F. 1974. Half saturation constant for nitrogen limited growth of
the green alga Selenastrum capricornutum. South Afr. J. Sci. 70(3):75-
76.
119
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algal assay, kinetics, limiting nutrients, nitrogen, continuous flow, algae,
Selenastrum capricornutum Printz, eutrophication.
The report is a brief discussion of results obtained from a continuous
flow algal assay conducted to determine the half-saturation content for nitro-
gen-limited growth of the test alga Selenastrum capricornutum Printz. Growth
of the algae was found to be limited by nitrogen; the calculated kinetics
value for Selenastrum capricornutum indicated a high affinity for nitrogen
during the growth period. The author felt a need to conduct further experi-
ments to test the limiting nutrient hypothesis and to aid in managing eutro-
phication problems. (Abstract-Leischman-EPA).
0135
Toerien, D. J. , and C. H. Huang. 1973. Algal growth prediction using growth
kinetic constants. Water Res. 7(11):1673-1681.
growth rates, kinetics, phosphorus, cultures, forecasting, time, eutrophica-
tion, biomass, nutrients, suspended solids, Selenastrum capricornutum Printz,
algal assay; bottle test, algae.
The growth kinetics under conditions of phosphorus limitation of Selenas-
trum capricornutum Printz, a green alga specified for use in algal bioassays,
were used to predict growth in batch cultures for varied specific conditions
of time and phosphorus concentration. These predictions compared very well
with actual batch culture growth studies. The predicted maximum cell concen-
tration for two different levels of phosphorus lay within values obtained in
different laboratories. The predicted maximum specific growth rates (u, ) were
either close to or just above actual laboratory data. The determination of
growth kinetics thus allows accurate prediction of the growth of planktonic
algae, a benefit in either algal bioassays or the solution of practical eutro-
phication problems. The growth kinetic constants of specific algae important
in eutrophication problems need to be determined in order to utilize poten-
tialities of prediction in the rational solution of these problems. (Ab-
stract-Author).
0136
Toerien, D. F. , and D. J. Steyn. 1974. The eutrophication levels of four
South African impoundments. Verh. Int. Ver. Limnol. 19(3):1947-1956.
120
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algal assay, eutrophication, wastewater, impoundment, AGP, limiting nutrients,
nitrogen, phosphorus, nutrients, algae, Anabaena circinalis, Selenastrum
capricornutum.
Algal assays were used to determine eutrophication levels of four South
African dams, Hartbeespoort, Rietvlei, Roodeplaat, and Vaal. The algal growth
potential (AGP) was measured for the test alga Selenastrum capricornutum.
Identification of the limiting nutrients; indication of stimulation from
additions of wastewater; and removal of nutrients by dilution with high qual-
ity water, were determined from the studies. The authors concluded that all
four impoundments were eutrophic and algal growth was stimulated by the waste-
water additions on a linear relationship. Nitrogen and phosphorus were the
primary and secondary limiting nutrients; the AGP increased with increased
concentrations of nitrogen. The removal of nutrients from the waters tested
was thought to be the potential method to control eutrophication. (Abstract-
Leischman-EPA).
0137
Toerien, D. F. , K. L. Hyman, and M. J. Bruwer. 1974. Comparison of the algal
bioassay responses of Selenastrum capricornutum and Microcystis
aeruginosa. South Afr. J. Sci. 70(3):75.
algal assay, eutrophication, bottle test, AGP, impoundment, algae, Selenastrum
capricornutum, Microcystis aeruginosa.
A brief discussion of results obtained from comparisons of growth rates
(AGP) of two test algae, Selenastrum capricornutum and Microcystis aeruginosa,
used in an algal assay for waters of Hartbeespoort Dam. Very significant
correlation coefficients to growth were found between the two algae from the
results. The authors found that the response of Selenastrum capricornutum can
be related to the growth of Microcystis aeruginosa. Summarized regression
analyses are given. (Abstract-Leischman-EPA).
0137-a
Toerien, D. F. , K. L. Hyman, and M. J. Bruwer. 1975. A preliminary trophic
status classification of some South African impoundments. Water S.A.
121
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algal assay, bottle test, limiting nutrients, nitrogen, phosphorus, impound-
ment, eutrophication, AGP, algae, Selenastrum capricornutum.
A preliminary trophic status classification of 98 major South African
impoundments was constructed using algal bioassay procedures to determine the
algal growth potential and to identify the growth limiting nutrients in each
impoundment. The classification procedure embodied a technique whereby the
growth potential of nitrogen fixing algae was accounted for. Although the
classification is based on a single parameter, it appears to provide a useful
guide to the trophic status of these impoundments. Approximately 50 percent
of the impoundments were low in plant nutrients, 11 percent were heavily
eutrophied and the rest were intermediate. Phosphorus was the most frequent
algal growth limiting nutrient in non-eutrophied impoundments and nitrogen the
most frequent in highly eutrophied ones. Because of the fixation of atmo-
spheric nitrogen by certain blue-green algae, the control of nitrogen appears
to have limited value in eutrophication control. Limiting the access of
phosphorus to impoundments seems to offer a long-term solution to eutrophica-
tion problems. Urban-industrial development giving rise to nutrient high
effluents, seems to be the major cause of eutrophication although the role of
agriculture can not be negated. (Abstract-Author).
137-b
Toerien, D. F. , D. J. Steyn, and S. I. Kohlmeyer 1975. Nitrogen yield
coefficient of the green alga, Selenastrum capricornutum Printz. South
Afr. J. Sci. 71:217-218.
algal assay, growth kinetics, nitrogen, phosphorus, nutrients, limiting nutri-
ents, algae, Selenastrum capricornutum Printz, impoundment, wastewater.
Water samples from various impoundments were tested to determine eutro-
phication levels. The samples were enriched with sewage effluent and varying
concentrations of nitrogen. Yield coefficients for the test alga Selenastrum
capricornutum were calculated for different concentrations of nitrogen and
phosphorus; a growth curve was constructed. Tests conducted in the algal
culture medium resulted in a predicted growth curve that was closely related
to the curve from the actual test waters. The authors concluded that it was
possible to predict growth of Selenastrum capricornutum under nitrogen limited
conditions. (Abstract-Leischman-EPA).
122
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0138
Toerien, D. F. , C. H. Huang, J. Radimsky, E. A. Pearson, and J. Scherfig.
1971. Final report: Provisional algal assay procedures. Sanitary Engr.
Research Lab. SERL Report, Berkeley, Calif., SERL Report No. 71-6. 211
P-
continuous flow, algal assay, nutrients, bottle test, lake waters, limiting
nutrients, algae, Selenastrum capricornutum, rivers, wastewater.
Batch and continuous flow (chemostat) assays were investigated as part of
a joint industry-government sponsored, multi-laboratory effort to develop a
standardized algal assay procedure for nutrient level assessment. Assays were
conducted with Selenastrum capricornutum as a standard assay organism.
Batch culture assays were found to have a lower level of precision than
chemostat assays in the assessment of growth response as a function of nutri-
.^
ent concentration. The biomass parameter, maximum cell concentration, X, of
the batch assay generally responded to the nutrient concentration of the
samples, however, the chemostat biomass parameter, steady state cell concen-
tration, X1? always was found to be proportional to the nutrient concentration
of the samples.
The results of spiking tests with batch assays generally were inconclu-
sive with respect to identification of the growth rate limiting nutrient
whereas the results of spiking tests with chemostats indicated clearly the
growth rate limiting nutrient. It is recommended that batch type algal assays
be used only for crude screening or routine monitoring purposes and that the
chemostat should be used for the quantitative assessment of the algal growth
supporting properties of waters as well as for the development of kinetic
descriptions for nuisance algae and the rate limiting nutrients of concern.
A kinetic description of Selenastrum capricornutum indicated a low half
saturation constant, K , (the concentration of nutrient supporting one-half
the maximum growth rate) of about 5 |jg P/£ for phosphate phosphorus and a
yield coefficient, Y, that varied as a function of growth rate. A theoretical
model was proposed and evaluated which describes the varying yield coefficient
(the result of "excess" uptake) as a function of the growth rate (mean cell
residence time). The function was verified experimentally at a very high
statistical confidence level. The significance of these findings and their
application to the practical problem of eutrophication assessment is pre-
sented. (Abstract-Author).
123
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0139
Trainor, F. R., and L. E. Shubert. 1973. Growth of Dictyosphaerium,
Selenastrum, and Scenedesmus (Chlorophyceae) in a dilute algal medium.
Phycologia 12(l/2):35-39.
algal assay, growth rates, rivers, bottle test, algae, Scenedesmus,
Dictyosphaerium, Selenastrum, media.
Cultures were maintained in dilute laboratory media for weeks by frequent
transfer of cells. Single strains of Scenedesmus and Dictyosphaerium were
cultured in a medium with 28.5 mg/1 total inorganic salts, with growth at the
rate of 1 + doublings/day. Growth of a Selenastrum was compared in one ex-
periment. Bristol's medium was also used for comparison.
Routinely, an actively growing inoculum was used, sufficient to give an
initial cell concentration of approximately 2 X 10s cells/ml; this was ade-
quate for colorimeter readings. Diurnal light of 3200 lux intensity was used
throughout. Each day, after a determination of growth, the culture was trans-
ferred by dilution to the original cell number. Some cultures have been grown
in this way for several weeks, and a uniform daily increment in growth is
reported for 10-day experiments. Using this technique we record similar
growth rates in sterile-filtered river water used as a medium. Stock cultures
can be held in dilute media for several weeks, by first establishing a healthy
population, and then maintaining the culture under minimal conditions, usually
light at 160-215 lux. (Abstract-Author).
0140
U.S. Environmental Protection Agency. 1971. Algal Assay Procedure Bottle
Test. U.S. Environmental Protection Agency, Corvallis, Oregon. 82 p.
algal assay., bottle test, eutrophication, nutrients, trace elements, growth
rates, toxicity, pretreatment, inhibition, stimulation, PAAP, limiting nutri-
ents, wastewater, chlorophyll, standing crop, temperature, light intensity,
phosphorus, nitrogen, N:P ratios, experimental design, data interpretation,
algae, Selenastrum capricornutum, Microcystis aeruginosa, Anabaena flos-aquae.
This document is the product of more than two years of intensive research
at governmental, industrial, and university laboratories to develop a reliable
and reproducible algal assay. The "Bottle Test" of the Provisional Algal
Assay Procedure (PAAP), which was published by the Joint Industry/Government
124
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Task Force on Eutrophication in February, 1969, served as the starting point
for this effort. In improving and evaluating the test, each laboratory, of
the participating group of nine, followed the same research plan using algal
test species from a common source. This course of action identifies elements
of the test that were faulty or difficult or questionable. As such specific
problems were recognized, selected laboratories were assigned the task of
probing them and developing sound adjustments. To do this they followed
research plans developed jointly by all laboratories. As a result of this
massive effort, the Algal Assay Procedure:Bottle Test was refined sufficiently
to be used in connection with eutrophication and other algal production prob-
lems. (Condensed forward-Author).
0141
U.S. Environmental Protection Agency. 1974. Dillon Reservoir - Blue River
Study, Colorado. U.S. Environmental Protection Agency, Denver, Colorado.
S & A / TIB - 28. 35 p.
reservoirs, eutrophication, wastewater, nutrients, rivers, algal assay, molyb-
denum, growth rates, algae, Selenastrum capricornutum, phosphorus, bottle
test.
Studies of the Dillon Reservoir - Blue River area involved algal assays
to determine nutrient levels; the effects of nutrient loadings from wastewater
discharge on algal productivity; and the possible effects, if nutrient load-
ings increased. The algal growth potential was measured for the alga Sele-
nastrum capricornutum in laboratory tests. High levels of molybdenum were
caused from mining activities; the algal assay was used to determine effects
of molybdenum on the growth of Selenastrum capricornutum in the waters.
(Abstract-Lei schman-EPA).
0142
U.S. Environmental Protection Agency. 1974. Biostimulation characteristics
of wastes and receiving waters of the Snake River basin. National Field
Investigations Center - Denver, Colorado and U.S. Environmental Protec-
tion Agency, Seattle, Washington. 49 p.
reservoirs, rivers, water quality, wastewater, nutrients, stimulation, phos-
phorus, feedlot runoff, algae, Selenastrum capricornutum, algal assay.
125
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Studies were designed to assess the characteristics of wastewater being
discharged into the Snake River basin. Algal assays were used to determine
the nutrient levels required to stimulate algal production; to provide infor-
mation that would aid in the removal of nutrients from the waters; and to add
further information pertaining to the development of discharge permits.
Research was conducted in the laboratory and field. (Abstract-Leischman-EPA).
0143
U.S. Geological Survey. 1975. Bioassay: Algal growth potential (AGP), p.
289-293. In P. E. Greeson, T. A. Ehlke, G. A. Irwin, B. W. Lium, and K.
V. Slack [eds.], Methods for collection and analysis of aquatic biologi-
cal and microbiological samples, Ch. A4, Book 5, U.S. Geological Survey,
Washington, D. C.
AGP, water quality, limiting nutrients, electronic particle counter, algal
assay, pretreatment, algae, Selenastrum capricornutum.
The report is a modification of the US EPA 1971 Algal Assay procedure
using the algal assay and the algal growth potential parameter. Methodology,
apparatus, reagents, collection, analysis, calculations, appendix and refer-
ences given. (Abstract-Leishman-EPA).
0144
Venkataraman, G. S. , and K. V. Natarrajan. 1958. Molybdenum requirements of
Selenastrum westii. Current ,Sci. 27(11):454-456.
molybdenum, growth rates, toxicity, trace elements, bottle test, algae, Sele-
nastrum westii, Scenedesmus obliquus, algal assay.
The report is of a study conducted to determine the molybdenum require-
ments for the alga Selenastrum westii. The authors describe the methods
followed in the experiment and the results, in terms of growth rates. Sele-
nastrum westi i was found to have a very low molybdenum requirement, as com-
pared to the requirement for Scenedesmus obliquus. The level of molybdenum
that was inhibitory to Selenastrum westii was also found to be less than the
level for Scenedesmus obliquus. (Abstract-Leischman-EPA).
0144-a
Vyas, H. 1975. The effects of dyes on environment, p. 251-269. _In Proc.
Third Annual Poll. Control Conf. Water and Wastewater Equip. Manuf.
126
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Assoc. Industry Solutions '75, Air-Water-Noise-Solid Waste, Ann Arbor
Sci. Mich.
dyes, pollution, toxicity, heavy metals, wastewater, anaerobic, aerobic,
inhibition, stimulation, algal assay, bottle test, algae, Selenastrum
capricornutum, fish, Pimephales promelas.
An attempt has been made in this paper to present some of the environ-
mentally related studies that ADMI has sponsored in the past. These studies
represent only the thin edge of the wedge. There is much left to be done,
however, we are very encouraged by the results obtained so far which seem to
show that dyes, for the most part, do not present serious problems to the
environment.
In our continuing quest for greater understanding of the role of dye in
the ecosystem, ADMI is currently directing a study, partly funded by EPA, to
investigate treatability of dying wastes by various biological and physico-
chemical treatments. This work is being carried out at the North Carolina
State University at Raleigh, and the University of North Carolina at Chapel
Hill. As a part of this project, a survey of textile mill effluents will be
carried out for selected heavy metal contents, TOC and benzidine. (Summary-
Author).
0145
Walmsley, R. D. , and P. J. Ashton. 1977. Algal growth potential and the
succession of algae in a nitrogen-limited impoundment. South Afr. J.
Sci. 73(5):151-152.
algal assay, bottle test, impoundment, AGP, nutrients, n.itrogen, phosphorus,
succession, limiting nutrients, algal blooms, algae, Anabaena flos-aquae,
Selenastrum capricornutum, phytoplankton, Anabaena circinalis, Volvox rous-
seletii, Microcystis aeruginosa Kutzing, chlorophyll a, standing crop.
An algal assay was used to test waters from the Rietvlei Dam and to
determine the potential of the waters to support algal growth. Two test
species were used, Selenastrum capricornutum and Anabaena flos-aquae. AGP
values were calculated; standing crop was estimated by analysis of chlorophyll
a. Results showed that phosphorus was always abundant and available to algal
growth; nitrogen was in limiting supply. The AGP values for the two test
algae were found to relate to algal blooms and succession within the impound-
ment. The nitrogen supply was thought to be the governing factor of succes-
127
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sion of the algae. Although the results yielded high AGP values, the authors
did not conclude that the values meant the development of large algal blooms.
The authors felt this indicated limitations involved with using assays to
predict algal growth. (Abstract-Leischman-EPA).
0146
Walmsley, R. D. , and D. F. Toerien. 1975. Algal bioassays and nitrogen
fixation. South Afr. J. Sci. 7(10):313-314.
nitrogen fixation, algal assay, bottle test, impoundment, limiting nutrients,
AGP, phosphorus, nutrients, algae, Selenastrum capricornutum, Anabaena flos-
aquae (Tischer1s-strain A37), eutrophication.
Studies were conducted in order to understand results from algal assays
of test waters where nitrogen is the primary limiting nutrient and phosphorus
the secondary limiting nutrient. Comparisons of two test algae, Selenastrum
capricornutum a green, non-nitrogen fixing alga; and Anabaena flos-aquae a
blue-green, nitrogen fixing alga, and the respective algal growth potential
values are given from tests of waters of Rietvlei Dam. (Abstract-Leischman-
EPA).
0146-a
Walmsley, R. D. , and D. F. Toerien. 1977. The summer condition of three
eastern Transvaal reservoirs and some considerations regarding the as-
sessment of trophic status. J. Limnol. Soc. South Afr. 3(2):37-41.
algal assay, bottle test, nutrients, nitrogen, phosphorus, chlorophyll a,
algae, Selenastrum capricornutum Printz.
Summer temperature, oxygen and light penetration profiles in the Da Gama,
Klipkopje and Witklip Reservoirs are presented. Analysis of waters from
different depths showed the presence of chemical stratification despite the
absence of clear thermal stratification. An algal bioassay procedure also
confirmed the presence of increasing quantities of available nutrients with
depth in all three reservoirs. The significance of the results in assessing
trophic status is discussed and it is concluded that for South African im-
poundments, an anaerobic hypolimnion is not exclusively associated with eutro-
phy. (Summary-Author).
128
-------
0147
Waimsley, R. D. , W. E. Scott, and D. F. Toerien. 1975. A laboratory evalua-
tion of a dye for the control of algal growth. Water SA 1(2):90-92.
dyes, growth rates, algal assay, bottle test, chlorophyll a, light penetra-
tion, algae, Selenastrum capricornutum, Anacystis m'dulans.
The principle of the use of a dye for the control of algal growth was
investigated at the laboratory scale. The commercial dye employed was shown
to have little effect on the growth rates of Anacystis nidulans and Sele-
nastrum capricornutum in the laboratory at concentrations above the recom-
mended doses. Experiments using 200 £ tanks also substantiated this lack of
effect on mixed phytoplankton populations. It is suggested that a dye with
absorption characteristics closer to those of algal pigments might be more
effectual for algal growth control. (Abstract-Author).
0148
Walmsley, R. D. , D. F. Toerien, and D. J. Steyn. 1978. An introduction to
the limnology of Roodeplaat Dam. J. Limnol. Soc. South Afr. 4(1): 35-52.
impoundment, eutrophication, nutrients, oxygen, AGP, nitrogen, phosphorus,
chlorophyll a, phytoplankton, Potomogeton, Microcystis, Microcystis aeru-
ginosa, toxicity, Melosira granulata, algae, Selenastrum capricornutum, algal
assay, diatoms.
A two-year limnological survey between 1973 and 1975 showed that the
Roodeplaat Dam is a warm monomictic impoundment which has stable thermal
stratification during summer. Oxygen supersaturation of the surface waters
and anaerobic conditions in the hypolimnion were encountered during summer.
Water transparency was seasonal and related to existing phytoplankton popula-
tions and allochthonously-introduced clay material. Chemical stratification
during summer was prominent and appeared to be caused not only by mineraliza-
tion processes in the hypolimnion, but also by dilution in the surface layers
during summer flooding. Because of the morphometry of the impoundment and the
fact that sewage effluents are discharged into the major inflow, considerable
horizontal variation in most chemical and biological characteristics was
found. On the basis of the nutrient content of the waters and the magnitude
of the phytoplankton populations observed, Roodeplaat Dam may be classified as
a highly eutrcphic water-body. (Summary-Author).
129
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0149
Walmsley, R. D. , D. F. Toerien, and D. J. Steyn. 1978. Eutrophication of
four Transvaal dams. Water SA 4(2): 61-75.
rivers, impoundment, eutrophication, wastewater, nutrients, algal assay, AGP,
algae, Selenastrum capricornutum.
A two year study of river waters in the catchment areas of the Buffels-
poort, Hartbeespoort, Rietvlei and Roodeplaat Dams was conducted. Sources of
eutrophying elements were detected by chemical analyses and algal bioassays.
The degree of enrichment of each impoundment was compared by means of phos-
phorus surface loading rates. Treated sewage effluents which are discharged
into the inflow waters of the Hartbeespoort, Rietvlei and Roodeplaat Dams are
the main cause of their present enriched condition. (Abstract-Author).
0150
Walton, C. P., and G. F. Lee. 1972. A biological evaluation of the molyb-
denum blue method for orthophosphate analysis. Verh. Int. Ver. Limnol.
18(2):676-684.
molybdenum, orthophosphate, lake waters, algal assay, bottle test, phosphorus,
limiting nutrients, algae, Selenastrum capricornutum.
A study has been conducted on the relationship between the results of the
molybdenum blue method for measuring soluble orthophosphate in natural waters
and the soluble phosphate concentrations present in natural waters which are
readily available for algal growth under standardized laboratory culture
techniques. The green alga Selenastrum capricornutum, was used as the test
organism. The lake waters tested were fortified with the nutrients contained
in the Provisional Algal Assay Procedure media such that the growth of the
test organism would be limited by the amount of phosphorus available in the
test water. It was found that there was no discrepancy between the amounts of
growth of the test organism in the Lake Mendota water, sediment extracts, and
algal detritus extracts predicted by the molybdenum blue soluble orthophos-
phate and the growth of the test organism in the test media with a corres-
ponding amount of soluble orthophosphate added. (Summary-Author).
130
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0150-a
Weiss, C. M. 1969. Batch culture tests: Effects of inoculum age and micro-
nutrients on growth of Selenastrum in PAAP medium. Univ. North Carolina,
Chapel Hill, N.C. Interim Report 1. FWPCA 16010 DQT.
algal assay, algae, Selenastrum, inoculum age, PAAP, media, growth rates,
micronutrients, bottle test.
Experiments were designed and carried out to resolve the following ques-
tions:
a) To what extent does algal inoculum age affect viability and nutrient
uptake on subculturing into PAAP medium.
b) Is it possible to maintain a stock algal culture in steady state
conditions by withdrawing a constant volume of liquids from culture
flasks, and replacing with an equivalent volume of fresh medium.
The overall objectve is to determine the optimum pre-culture conditions
necessary to obtain a viable inoculum of Selenastrum with minimum nutrient
content in the cells, for use in assaying lake waters.
This report deals with the first half of a series of experiments, involv-
ing studies on the effect of aging cultures for 5-21 days, and of maintaining
a steady state culture, on subsequent viability and nutrient uptake, as deter-
mined by subculturing in different dilutions of the PAAP medium (October 1969-
version). Good replication and a uniformity of trends in regard to the basic
features of algal growth and nutrient uptake have confirmed previous results
obtained by Weiss and other workers.
The above experiments were conducted with 50 ml liquid volumes in 125 ml
culture flasks with an initial algal cell concentration of 103 cells/ml.
(Modified Summary-Author).
0151
Weiss, C. M. 1970. Batch culture tests: Effects of inoculum age and micro-
nutrients on growth of Selenastrum in PAAP medium. Univ. North Carolina,
Chapel Hill, N.C. Interim Report 2. FWPCA 16010 DQT.
inoculum age, PAAP, media, growth rates, micronutrients, carbon, nitrate,
bottle test, algal assay, algae, Selenastrum.
131
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This report presents the results of experiments which replicate the
experiments reported in Interim Report #1 in which the effect of age of cul-
ture on the growth of Selenastrum in PAAP medium was examined. Growth re-
sponse was compared at 800 fc and 400 fc in 10 and 100% PAAP media with and
without micronutrients. Detailed examination of the results of the second age
of culture experiment is made and statistical comparisons of the two experi-
ments is presented. The conclusions drawn indicate that age of culture be-
tween 7 and 21 days has no statistically significant effect on growth. No
significant effect of light intensity between 800 and 400 fc on algal growth
could be demonstrated but the higher light intensity did appear to produce
significantly greater carbon production in 100% PAAP. The rate of N03-N
uptake was considerably higher in the +m PAAP than in -m PAAP and thus appears
to be dependent on the presence of micronutrients. The presence of micro-
nutrients in PAAP medium is indicated as highly desirable since in all compar-
ative experiments micronutrients enhanced the growth of algae, carbon syn-
thesis and N03-N uptake. (Summary-Author).
0152
Weiss, C. M. 1970. Batch culture tests: Effects of inoculum age and micro-
nutrients on growth of Selenastrum in PAAP medium. Univ. North Carolina,
Chapel Hill, N.C. Interim Report 3. FWPCA 16010 DQT.
growth rates, inoculum age, media, bottle test, algal assay, algae, Sele-
nastrum, nutrients.
Studies were conducted in two laboratories, University of Wisconsin and
University of North Carolina, on the effects of inoculum age and media concen-
trations on specific growth rates of Selenastrum. A comparison of the two
laboratory's conclusions is outlined in this report. Tables of specific
growth rates and final yields are given. (Abstract-Leischman-EPA).
0153
Weiss, C. M. 1970. Effect of pH on precipitation and resolubilization of
phosphate-phosphorus in algal cultures. Univ. North Carolina, Chapel
Hill, N.C. Interim Report 4. FWPCA 16010 DQT.
pH, phosphorus, media, culture age, phosphate, precipitation, nutrients,
resolubilization, algae, Selenastrum, algal assay, bottle test.
132
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The report is of a study conducted to determine the effects of pH on the
precipitation and resolubilization of phosphorus within algal cultures.
Cultures of Selenastrum used in the tests, ranged from six to 34 days of age.
Maximum removal of P04=P from the media was found to occur around day seven of
incubation. It was found that adjustment of the pH by acidification did not
greatly increase the P04=P concentration in the media, compared to samples
untreated; however, the increase of P04=P in cultures older than 13-27 days
was greater after acidification. The author concluded that in comparing sam-
ples that were acidified to samples unacidified, it was found that some in-
crease in dissolved P04=P concentrations did occur following acidification in
the majority of samples. (Abstract-Leischman-EPA).
0154
Weiss, C. M. 1970. Batch culture tests: Effects of inoculum size on growth
of Selenastrum in PAAP medium. Univ. North Carolina, Chapel Hill, N.C.
Interim Report 5. FWPCA 16010 DQT.
inoculum, growth rates, PAAP, media, carbon, phosphate, nitrate, bottle test,
algal assay, algae, Selenastrum.
This report presents the results of an experiment designed to examine the
effect of inoculum size on the growth of Selenastrum in +m and -m 100% PAAP
media at 800 fc. Cultures were inoculated with 1,000, 10,000 and 50,000
cells/ml and measurements of algal numbers, algal carbon content, P04-P and
N03-N concentrations were made after 5, 7, 9 and 14 days incubation. The
results indicate that inoculum size has no effect on algal yield in either +m
or -m PAAP after 14 days incubation. Higher yields are produced in the +m
PAAP, however, than in -m. The P04-P uptake rate was found to increase with
increasing inoculum size and was, in general, greater in +m PAAP than in -m.
Although inoculum size had little effect on the N03-N uptake rate, the +m PAAP
medium greatly enhanced the rate of N03-N utilization. The inoculum size had
little effect on carbon synthesized in either +m or -m PAAP but there was a
large difference in the amount of carbon synthesized by cells grown in the two
media. (Summary-Author).
133
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0155
Weiss, C. M. 1970. Factors affecting growth of Selenastrum in batch cul-
tures: Removal of samples, surface-volume ratio of media, carbon limit-
ation. Univ. North Carolina, Chapel Hill, N.C. Interim Report 6. FWPCA
16010 DQT.
algal assay, bottle test, growth rates, surface:volume ratios, carbon limit-
ation, PAAP, algae, Selenastrum.
Three experiments were conducted to investigate the effects of various
procedural aspects on the growth of Selenastrum in the Provisional Algal Assay
Procedure. Experiment I contrasted the growth of Selenastrum in cultures from
which samples were removed periodically with the growth in cultures from which
no samples were taken. Experiment 2 was conducted to determine the effect of
surface-volume ratio, as reflected by the various combinations of flask size
and initial culture volume as well as the removal of different size samples,
on the growth of Selenastrum. The effect of surface-volume ratio, in terms of
the potential carbon limitation possibly imposed upon the cultures of low
surface-volume ratio, was investigated in Experiment 3. In this experiment,
differences in growth attributable to the use of aluminum foil vs. foam plas-
tic plugs because of gas exchange limitations were also studied.
The results indicate that the surface-volume ratio of the culture exerts
a profound influence on the growth of Selenastrum in 100% PAAP. The increased
surface-volume ratio produced by the periodic removal of samples resulted in a
higher algal yield. Likewise, variations in growth were observed in different
flask sizes which reflected the differences in surface-volume ratio among the
flasks.
It was found that in 10% PAAP neither surface-volume ratio nor the addi-
tion of double the normal amount of NaHC03 affected the total yield of algae
indicating that carbon limitation did not occur in this medium.
Little difference in growth was noted between cultures grown with foil
caps and those stoppered with foam plastic plugs. (Summary-Author).
0156
Weiss, C. M. 1970. Batch culture tests: The effects of surface volume ratio
on growth of Selenastrum in PAAP media of varying strengths. Univ. North
Carolina, Chapel Hill, N.C. Interim Report 7. FWQA 16010 DWT.
134
-------
PAAP, media, surface:volume ratios, carbon, nutrients, algae, Selenastrum,
bottle test, algal assay.
Cultures of Selenastrum were grown in a variety of strengths of PAAP
medium under three different surface volume ratios. All other ambient condi-
tions were held constant. Algal number and total carbon were used for indices
of algal yield. A proportional relationship between yield and nutrient
strength from 0 to 100 percent PAAP was attained after 21 days incubation in
cultures with a surface volume ratio of 1.04. At lower surface volume ratios
proportionality was evident only at reduced medium strengths. In cultures
with a surface volume ratio of 0.22 the correspondence between yield and
nutrient strength was maintained to no more than 40% of the October 1969 PAAP.
It was suggested that either a medium strength of 30 percent of the
October 1969 PAAP or that a surface volume ratio of at least 1.0 be used in
subsequent PAAP studies. A possible carbon limitation effect is suggested by
the results of these experiments. (Abstract-Author).
0157
Weiss, C. M. 1970. Batch culture tests: The effects of the micronutrient
history of algal stock cultures on subsequent growth in media with or
without micronutrients. Univ. North Carolina, Chapel Hill, N.C. Interim
Report 8. FWPCA 16010 DQT.
media, PAAP, micronutrients, phosphorus, nitrogen, carbon, phosphate, nitrate,
growth rates, algae, Selenastrum capricornutum, bottle test, algal assay.
Stock cultures of Selenastrum which had been carried consistently in one
type of medium (PAAP with or without micronutrients) were transferred to both
media types. After 9 days incubation, cells from each of'these cultures were
also transferred to both -M and +M PAAP media. These cultures were incubated
14 days, at which time measurements were made of P04-P and N03-N uptake,
organic carbon and the growth in cell number.
The results indicated that no relationship between the micronutrient
history of the culture and the growth in cell number, the amount of carbon
assimilation, P04-P or N03-N uptake. The growth in cell number, the amount of
carbon assimilation and the N03-N uptake in the -M medium were, however,
consistently lower than in the +M medium, regardless of the history of the
stock algae. (Abstract-Author).
135
-------
0158
Weiss, C. M. 1970. Batch culture tests: The exchange of phosphorus between
algal cells and media as related to the age of culture. Univ. North
Carolina, Chapel Hill, N.C. Interim Report 9. FWPCA 16010 DQT.
phosphorus, carbon, nitrogen, PAAP, bottle test, algal assay, media, nutri-
ents, algae, Selenastrum capricornutum.
Selenastrum were cultured in 100% (October 1969) PAAP media for 35 days.
At intervals throughout the incubation period samples were taken and analyzed
for P04-P, N03-N and organic carbon content, as well as growth in cell number.
At each sampling a portion of the sample was inoculated into 10% PAAP media.
These cultures were analyzed at 14 days for growth in cell number and organic
carbon content.
Of primary interest in this work was: (1) the phosphorus content of the
algal cells (cultured in 100% PAAP) as related to the age of the algae, (2)
and the effects of the algal phosphorus content on the growth of these cells
when inoculated into fresh medium. The results indicated a relatively high
level of phosphorus in the algal cells of the 100% PAAP cultures during the
first three days of incubation. The level dropped sharply while the cells
were in the log phase of growth, to a low at about 7 days incubation. The
algal phosphorus content remained constant, thereafter. Cells of 3 to 35 days
of age, when inoculated into 10% PAAP medium, showed no significant differ-
ences in growth, measured in terms of algal number and organic carbon at 14
days incubation. (Abstract-Author).
0159
Weiss, C. M. 1970. Influence of nitrilotriacetate (NTA) on eutrophication i_n
situ and batch assays. Univ. North Carolina, Chapel Hill, N.C. Interim
Report 11. FWQA 16010 DQT.
nitrilotriacetate, NTA, stimulation, carbon-14, algal assay, bottle test,
wastewater, nitrogen, carbon, phosphorus, lake waters, NAAM, media, impound-
ment, algae, Selenastrum capricornutum, phytoplankton.
The potential of nitrilotriacetate (NTA) to act as an algal growth stimu-
lant was evaluated by i_n situ carbon-14 productivity determinations and batch
assay according to the PAAP procedure. The NTA in pure chemical form as well
as following passage through secondary and tertiary sewage treatment was
136
-------
tested in comparison with nitrogen and carbon provided from other chemical
sources in amounts approximately equivalent to that found in the NTA molecule.
In addition to untreated controls, the lake waters and NAAM media (0.3 PAAP
media of October 1969), one test media included a phosphorus addition to the
lake waters.
The i_n situ and batch assays were made on three North Carolina impound-
ments over the period of July to October 1970. Each lake water was used for
four test series, of the ui situ and batch assays respectively. The i_n situ
experiments to determine effects of NTA on productivity as determined by
carbon-14 uptake were of four hour duration and the sets of light-dark bottles
were hung at a depth which provided 50% of surface illumination. Batch assays
used 0.45 (j membrane filtered water reseeded with Selenastrum capricornutum at
10s cells/ml. Growth responses in the various test media were followed by
cell counts through 21 days incubation at 24°C and 400 f.c. on an oscillatory
shaker at 110 oscillations/min. Dry weight of cell mass formed and carbon
synthesized were also determined at the end of the 21 day incubation period.
Changes in pH of the media were also followed through the growth period.
Evaluation of the i_n situ productivity results found that the presence of
NTA either in pure form or following passage through some level of sewage
treatment produced no change in productivity, carbon fixed (mg/l-hr-1), that^
exceeded the 90% confidence limits of the untreated lake water controls. The
only materials added to the lake water which resulted in productivity values
that were outside the confidence limits of the controls were the Na and K
carbonates and those on several tests resulted in productivity levels that
were lower than the controls.
The batch assays of NTA as an algal growth stimulant essentially con-
firmed the results of the productivity tests. No medium containing NTA either
in pure form or after passage through secondary or tertiary sewage treatment
levels resulted in growth, by any parameter used, which exceeded that of the
specific control in any significant manner. The phosphorus supplement used
for comparative purposes in this test, even though used at 1/5 the concentra-
tion originally specified resulted, in nearly every assay series, in growth
greater than the control, the nitrogen supplement or the NTA addition.
One interesting aspect of the productivity determinations was the finding
that if the productivity of the lake controls were compared -to the total
137
-------
phytoplankton count of the sample and total insulation for the period of
exposure there appeared to be indicated an optimum cell concentration and
insolation for the peak productivity values. (Summary-Author).
0160
Weiss, C. M. 1970. A statistical analysis of the effect on nitrilotriacetate
(NTA) and other algal nutrients on productivity i_n situ and algal assay
when tested in a randomized twelve block design. Univ. North Carolina,
Chapel Hill, N.C. Interim Report 12. FWQA 16010 DQT.
lake waters, nutrients, wastewater, NTA, nitrilotriacetate, nitrogen, carbon,
carbon-14, uptake, statistical methods, algal assay, bottle test, Selenastrum
capricornutum, stimulation, algae.
From the results of the preceding statistical analysis, it would appear
that the presence of nitrilotriacetate (NTA) on entering an aquatic environ-
ment through a sewage treatement plant could very well provide additional
nitrogenous nutrients which would be available for the support of algal
growth. It would appear that the sewage treatment process does break down the
NTA molecule but in doing so apparently makes more readily available the
nitrogen for cellular metabolism as compared to the pure chemical. The pure
chemical would appear to have no significant effect on algal growth as tested
in the batch assay procedure although it did appear to significantly stimulate
productivity as determined by the uptake of NaH14C03. (Conclusions-Author).
0161
Weiss, C. M. 1976. Evaluation of the algal assay procedure. U.S. Environ-
mental Protection Agency, Corvallis, Oregon. Ecological Research Series
EPA-600/3-76-064.
lake waters, algal assay, bottle test, impoundment, rivers, nutrients, pre-
treatment, water quality, chlorophyll a, limiting nutrients, algae,
Selenastrum capricornutum.
Evaluation of the algal assay bottle test and its relationship to the
trophic state or nutrient levels of surface waters was examined in 44 lakes,
impoundments and rivers in North Carolina in 345 separate assay sets. Of
particular concern was the evaluation of the significance of the pretreatment
procedure, autoclaving or filtration, upon growth of the reseeded alga in
relationship to the original water quality.
138
-------
A limnological data profile was developed for each of the bodies of water
sampled. A data processing procedure was used to establish the relationship
between water quality data and algal cell density, chlorophyll a and produc-
tivity.
The algal assay procedure provided an indication of limiting nutrient,
phosphorus, nitrogen or both. By clustering all samples of similar nutrient
limitation a basic nitrogen, phosphorus relationship emerged. When the ratio
of soluble inorganic nitrogen to total soluble phosphorus was greater than 13
the waters were phosphorus limited; when the ratio was in the range of 9-12
both nutrients were limiting; and when the ratio was below 8 nitrogen was
limiting.
The quantity of algal biomass grown in autoclaved or filtered samples,
without the addition of nutrient spikes provides an indication of the relative
trophic or nutrient level of the particular body of water, the former a mea-
sure of total growth potential and the latter ambient growth potential.
(Abstract-Author).
0162
Weiss, C. M. 1976. Field evaluation of the algal assay procedure on surface
waters of North Carolina, p. 29-76. J.n E. J. Middlebrooks, D. H.
Falkenborg, and T. E. Maloney [edsj, Biostimulation and nutrient assess-
ment, Ann Arbor Sci. , Mich.
algal assay, nutrients, nitrogen, phosphorus, lake waters, rivers, pond water,
impoundment, chlorophyll, pretreatment, growth rates, limiting nutrients, N:P
ratios, algae, Selenastrum capricornutum.
To establish relationships between assays and existing trophic levels of
water bodies, algal assays were made of 44 North Carolina water bodies. Cell
density decreased with inorganic nitrogen increase; chlorophyll remained
relatively constant; productivity reached a peak in middle concentration
ranges. In the growing season soluble nitrogen showed an inverse relationship
between cell number and nutrients. Phosphorus values varied but were higher
in winter. Chlorophyll showed a positive relationship to cell density, pro-
ductivity, and nitrogen. In the growing season there was a parallel increase
in organic nitrogen and soluble phosphorus and increased productivity; cell
density, and chlorophyll. Selenastrum capricornutum growth after autoclaving
or filtering water samples indicated total and ambient growth potentials
139
-------
respectively. Growth responses to added phosphorus or nitrogen compared to
growth with original nutrient levels indicated nitrogen:phosphorus ranges,
soluble nitrogen:phosphorus determinations following limitation of either or
both nutrients indicated that the ratio can define which nutrient is limiting
for a particular water body. The standard assay procedure should be modified
to limit the determination to biomass grown in reseeded samples after auto-
claving and filtration. Whether a sample is limited in growth potential by
the relative quantity of phosphorus or nitrogen can be determined from the
total soluble nitrogen:phosphorus ratio. (Abstract-Buchanan-Davidson-Wiscon-
sin WRSIC).
0163
Weiss, C. M. , and R. W. Helms. 1971. Inter!aboratory precision tests: An
eight laboratory evaluation of the Provisional Algal Assay Procedure
Bottle Test. National Eutrophication Research Program. U.S. Environmen-
tal Protection Agency, Corvallis, Oregon. 70 p.
algal assay, bottle test, media, nutrients, lake waters, PAAP, chlorophyll,
fluorescence, carbon, statistical methods, algae, Selenastrum capricornutum.
In order to establish the validity of an algal assay procedure for the
determination of algal nutrient levels in surface waters a suitable protocol
was designed and followed by eight laboratories. This group consisted of one
government laboratory, four university laboratories and three industrial
laboratories. The basic procedure was to evaluate by use of the "bottle" or
batch test the precision and reproducibil ity of the growth response of one
test organism, Selenastrum capricornutum, in four media varying nutrient
strength. The media was originally defined for the PAAP test and modified
slightly in subsequent evaluations. The test media of this experiment were
all dilutions of the PAAP medium.
In addition to following the growth of the test alga in the four "manda-
tory" media, several of the participating laboratories examined growth re-
sponse in the basic medium minus specific nutrient elements. The test proto-
col also permitted evaluation of growth controlling factors such as continuous
shaking versus handswirling, comparison of the use of the hemacytometer versus
Coulter Counter in determining cell numbers, and the levels of correlation'
between cell mass and cell number as determined by optical density, chloro-
phyll fluorescence and total algal carbon.
140
-------
The unusual nature of this evaluation in the numbers of laboratories
participating and magnitude of the test size has permitted the statistical
analysis to be developed to the extent that the results will set new guide-
lines for future assays of this type. (Summary-Author).
0164
Williams, P. H. 1975. Response of the Spokane River diatom community to
primary sewage effluent. M.S. thesis, East. Wash. State College, Cheney,
Wash. 38 p.
wastewater, species diversity, species composition, phosphate, nitrogen,
diatoms, ammonia, heavy metals, inhibition, pollution, organics, algae,
Selenastrum capricornutum Printz, algal assay.
This study examined the effects of the primary sewage effluent from the
City of Spokane, Washington on the diatom community in the Spokane River.
Both natural and artificial substrates were used at each of three sampling
stations, one above and two below the sewage outfall, to determine the impact
of the effluent on species composition, species diversity, and autotrophic
index. Water chemistry samples were taken concurrently with diatom collec-
tions and composited for use in determining the algal growth potential of the
water.
A total of 52 species in 18 genera of diatoms were identified. The flora
was dominated by Achnanthes spp. with this genus accounting for 63.6 percent
of the cells observed. Nine of the ten most abundant diatom taxa showed
statistically significant changes in abundance below the sewage discharge. In
each of these ten species there was a significant correlation between their
numbers and orthophosphate or ammonia nitrogen concentrations.
Mean autotrophic index values of 203 above and 1005 below the sewage
outfall indicate the effluent promoted an increase in heterotrophic biomass.
Results from algal assays using Selenastrum capricornutum Printz as a
test organism suggest that total algal growth above the discharge may have
been limited by phosphorus concentrations while below the effluent heavy metal
inhibition seemed to predominate.
In terms of individual species abundance, mean diversity indices, floris-
tic similarities between sampling stations, and mean autotrophic indices, the
diatom community at the last downstream station had begun to recover from the
effects of organic pollution introduced via the sewage. (Abstract-Author).
141
-------
0165
Won, W. D. , L. H. DiSalvo, and N. G. James. 1976. Toxicity and mutagenicity
of 2, 4, 6-trinitrotoluene and its microbial metabolites. Appl. Environ.
Microbiol. 31(4):576-580.
TNT, toxicity, mutagenicity, metabolites, algal assay, bottle test, algae,
Selenastrum capricornutum, copepods, oyster larvae.
TNT (2,4,6-trinitrotoluene) of explosive grade is highly toxic to the
freshwater unicellular green alga Selenastrum capricornutum, and to the marine
copepod Tigriopus caliform'cus, and the marine oyster larva Crassostrea gigas.
It is also mutagenic to Salmonella typhimurium. On the basis of mutagenic
assays carried out with a set of histidine-requiring strains of the bacterium,
TNT was detected as a frameshift mutagen that significantly accelerates the
reversion rate of a frameshift tester, TA-98. In contrast, the major micro-
bial metabolites of TNT appeared to be nontoxic and nonmutagenic. (Abstract-
Author).
0166
Wright, J. Jr., F. A. Camp, and J. Cairn, Jr. 1974. Preliminary algal bio-
assays to determine nutrients limiting algal productivity in Mountain
Lake, Virginia. ASB Bull. 21(2):92.
lake waters, nutrients, limiting nutrients, phosphate, nitrate, algal assay,
bottle test, algae, Selenastrum capricornutum, nitrogen, phosphorus.
Mountain Lake, the only natural lake in the unglaciated portion of the
Appalachian Mountains, is located at an altitude of 1200 m. (3970 ft) in an
undeveloped section of Giles Country, Virginia. It has a length of 1.2 km.
and a width of 0.4 km. with a surface area of about 100 acres. Proposed
development of areas immediately surrounding the lake prompted the authors to
conduct preliminary studies to evaluate the lake's algal growth potential by
determining (1) what nutrients were presently limiting algal productivity in
the lake, and (2) what concentrations of these nutrients were necessary to
cause a statistically significant increase in algal growth. Chemical analyses
conducted in the summer and fall of 1973 indicated relatively constant nitrate
and phosphate concentrations throughout the lake. Only a trace of phosphate
(as orthophosphate) was present while nitrate levels were approximately 0.3
142
-------
ppm. Previous researchers had reported low levels of nitrate and phosphates
throughout the year and characterized the lake as being oligotrophic. Assays
were conducted in the summer, fall and winter of 1973, using the alga Sele-
nastrum capricornutum as the assay organism. Nutrient spiking techniques were
generally those suggested in the Environmental Protection Agency's Algal Assay
Procedure Bottle Test. Phosphorus appeared to be the primary limiting nutri-
ent, and nitrogen became limiting when phosphorus was added in excess. (Ab-
stract Only-Author).
143
-------
2.0 Keyword Index
adenosine triphosphate
0006
0060-a
0062
aerobic
0026
0144-a
AGP
0014
0015
0016
0017
0017-a
0029
0029-a
0030
0030-a
0030-b
0032
0035
0055
0059-a
0060
0087
0124
0128-b
0128-c
0128-d
0130
0136
0137
0137-a
0143
0145
0146
0148
0149
agricultural runoff
0022-a
0027-a
agricultural watersheds
0050
aldrin
algae
see herbicides
0001
0001-a
0001-b
0002
0003
0003-a
0003-b
0003-c
0003-d
0003-e
0004
0005
0006
0007
0008
0008-a
0009
0009-a
0010
0011
0012
0013
0013-a
0013-b
0014
0015
0016
0016-a
0017
0017-a
0018
0019
0020
0020-a
0020-b
0020-c
0021
0022
0022-a
144
-------
algae (continued) algae
°059
0024 0059-a
0025 006°
0023
0024
0025
0026
0027 006°-b
0027-a
0027-b
0027-d
0027-e °065
0027-f O065'a
0028 °066
0028-a 0066-a
0029 °067
0029-a °068
0030 °069
0030-a 0070
0030-b °07'
0031 °072
0031-a 0073
0032 °074
0033 0075
0034 0076
0035 0077
0036 0078
0037 0079
0038 0080
nn-jq 0080-a
0040
0041 0083
0042 UUb-:5
°°« 0085
0044 °085
0044-a ««-
°086
OOS6-
4 0087
0048 °088
0049 °089
00509 0090
nnqo
0052
0094
-, uua^f
°095
0058 0098-b
145
-------
algae (continued) algae (continued)
0098-c 0134
0099 0135
0100 0136
0101 0137
0102 0138
0102-a 0139
0103
0104
0104-a
0103 0140
0104 0141
0142
0104-b 0143
0105 0144
0106 0144-a
0107 0145
0108 0146
0108-a 0146-a
0109 0147
0110 0148
0111 0149
0112 0150
OH3 0150-a
0113-a 0151
0114 0152
OH5 0153
0116 Q-,54
OH7 0155
0118 0156
0119 0157
OH9-a 0158
0120 0159
2
« 0161
Sill
o°]ff
U I £O
0126-a
0126-b °166
algal assay
028-a
0128 a 0002
0128-b ^003
0128 c onm-a
0128-d nnn^ h
01?q 0003-b
0 30 °°03-c
0 31 °003"d
0 32 °°°3-e
0 33 °°04
Ul66 0005
146
-------
algal assay (continued)
algal assay (continued)
0006
0007
0008
0008-a
0009
0009-a
0010
0011
0012
0013
0013-a
0013-b
0014
0015
0016
0016-a
0017
0017-a
0018
0019
0020
0020-a
0020-b
0020-c
0022
0022-a
0023
0024
0025
0026
0027
0027-a
0027-b
0027-c
0027-d
0027-e
0027-f
0028
0028-a
0029
0029-a
0030
0030-a
0030-b
0031
0031-a
0032
0033
0034
0035
0036
0037
0038
0039
0040
0041
0042
0043
0044
0044-a
0045
0046
0047
0048
0049
0050
0050-a
0051
0052
0052-a
0053
0054
0055
0056
0057
0058
0059
0059-a
0060
0060-a
0060-b
0061
0062
0063
0064
0065
0065-a
0066
0066-a
0067
0068
0069
0070
0071
0072
0073
0074
0075
0076
0077
0078
147
-------
algal assay (continued) algal assay (continued)
0079 0120
0080 0121
0080-a 0122
0081 0123
0082 0124
0083 0125
0084 0126
0085 0126-a
0085-a 0126-b
0086 0126-d
0086-a 0127
0087 0128
0089 0128-a
0090 0128-b
0091 0128-c
0093 0128-d
0094 0129
0095 0130
0096 0131
0097 0132
0098 0133
0098-a 0134
0098-b 0135
0098-c 0136
0099 , 0137
0100 0137-a
0101 0137-b
0102 0138
0102-a 0139
0103 0140
0104 0141
0104-a 0142
0104-b 0143
0105 0144
0106 0144-a
0107 0145
0108 0146
0108-a 0146-a
0109 0147
0110 0148
om 0149
0112 0150
0113 0150-a
01 3-a 0151
04 0152
S ? 0153
0116 0154
0117 0155
0118 0156
01 9 0157
0119-a 0158
148
-------
algal assay (continued)
Anabaena circinalis (continued)
0159
0160
0161
0162
0163
0164
0165
0166
algal blooms
algicidal
ammonia
Amphora
Anabaena
0066
0086-a
0145
0002
0027-b
0048
0095
0097
0027
0027-b
0028
0040
0075
0124
0164
0050-a
0027-a
0027-b
0060-b
0095
0099
Anabaena circinalis
0038
0061
0084
0136
0145
Anabena cylindrica
0067
Anabaena flos-aquae
0001
0003-a
0003-b
0003-c
0003-d
0006
0018
0020-b
0027
0045
0052
0057
0067
0072
0074
0082
0086-a
0087
0094
0096
0100
0101
0109
0114
0118
0128-a
0.129
0133
0141
0145
Anabaena rios-aquae (Lyngb.)
DeBrebisson
0116
Anabaena flos-aquae (Tischer's-
strain A37)
0146
149
-------
Anabaena scheremetievi
0038
Anabaena splroides
0038
Anabaena wisconsinense
0038
Anacystis
0066
Anacystis m'dulans
0117
0118
0133
0147
anaerobic
0144-a
Ankistrodesmus
0028
0083
0119
Ankistrodesmus braunii
0004
0118
Ankistodesmus falcatus
0004
0014
0015
0017
0027-f
0029
0030
0030-b
0032
0063
Ankistrodesmus falcatus (Corda) Ralfs.
0028-a
Aphanizomenon flos-aquae
0091
Asterionella formosa
ATP
0060-a
see adenosine
triphosphate
atrazine
0050-a
Bacillariophyceae
0122
biodegradation
0009-a
0027-b
bioindicators
0009
biomass
0135
biosynthesis
0021
bottle test
0001-a
0002
0003-a
0003-b
0003-c
0003-d
0005
0007
0008
0009-a
0011
0012
0013
0013-a
150
-------
bottle test (continued)
bottle test (continued)
0013-b
0014
0015
0017
0018
0019
0020-a
0020-b
0022-a
0023
0025
0027-c
0027-d
0027-e
0029
0035
0038
0043
0044
0044-a
0045
0046
0048
0051
0052
0052-a
0053
0054
0056
0057
0059
0059-a
0060-b
0061
0063
0064
0065-a
0066-a
0069
0071
0072
0074
0075
0076
0077
0078
0079
0080-a
0081
0082
0085
0086-a
0087
0090
0091
0093
0094
0095
0096
0097
0098
0098-a
0102
0103
0104
0105
0109
0111
0112
0113
0113-a
0114
0115
one
0123
0125
0126
0126-b
0128-a
0128-b
0129
0131
0132
0135
0137
0137-a
0138
0139
0140
0141
0144
0144-a
0145
0146
0146-a
0147
0150
0150-a
0151
0152
151
-------
bottle test (continued)
carbon dioxide
buffers
cadmium
0153
0154
0155
0156
0157
0158
0159
0160
0161
0163
0165
0166
0005
0068
0002
0013
0034
0060-a
0078
0005
0064
0085
0133
carbon limitation
0155
carbon-14
0001
0042
0052
0109
0159
0160
carotenoids
0021
0089
CCCP
Calothrix
0089
carbon
0004
cell quota
0007
0027-c
0028-a
0041
0058
0068
0071
0072
0075
0133
0151
0154
0156
0157
0158
0159
0160
0163
chelation
0029-a
0095
0096
chemical analysis
0027-d
0032
chemical flocculation
0030-a
Chlamydomonas
0130
152
-------
Chlamydomonas reinhardtil
0060-a
0117
0118
Chlorella
0006
0015
0028
0028-a
0032
0050-a
0063
0089
0099
0130
Chlorella ellipsoidea
0083
Chlorella fusca
0004
Chlorella fusca (strain 211-8b)
0021
Chlorella homosphaera
0014
0015
0017
0027-f
Chlorella kesslerii
0004
Chlorella oval is Butcher
0119-a
Chlorella pyrenoidosa
0027
0027-b
0042
0067
Chlorella pyrenoidosa
(continued)
0083
0084
0086
0087
0118
0133
Chlorella pyrenoidosa (211-8k)
0004
Chlorella stigmatophora
0013-b
Chlorella vulgaris
0083
0098-b
0117
chlorelline extracts
0050-a
Chlorophyceae
0122
chlorophyl1
0003-a
0003-c
0003-d
0006
0016-a
0017-a
0022-a
0028-a
0029-a
0031
0031-a
0046
0060-a
0060-b
0086
0095
0108-a
0118
0140
153
-------
chlorophyll (continued)
0145
0146-a
0147
0148
0161
0162
0163
chlorophyll a
0003-a
0003-c
0003-d
0006
0016-a
0017-a
0022-a
0028-a
0031
0031-a
0060-a
0060-b
0086
0095
0108-a
0118
0145
0146-a
0147
0148
0161
chlorophyll b
0118
chromium
0013
Cladophora
0026
0027
0027-c
0027-d
0027-e
0131
classification
0092
0120
coastal waters
0122
cobalt
0013
0061
Cocconies
COD
0050-a
0031-a
0120
Coelastrum proboscideum
0004
colchicine
0050-a
competition
0086-a
continuous flow
0008-a
0041
0049
0052
0052-a
0060-a
0068
0074
0075
0086
0099
0102
0102-a
0104-a
0104-b
154
-------
continuous flow (continued)
0105
0106
0107
0113
0127
0128
0134
0138
copepods
copper
0165
0002
0013
0013-b
0027-b
0050
0060-a
0078
culture age
0153
cultures
0001-b
0108
0119-a
0122
0126-c
0135
Cyanophyceae
0122
cyclohexylamine
0037
Cyclotella
0118
cyclotrimethylene trinitramine
0003-a
Cylindrospermum
0067
Cymbella
0050-a
dacthal
see herbicides
data interpretation
0003
0079
0140
DCMU
0004
detergents
dialysis
diatoms
0023
0033
0048
0096
0100
0060-a
0127
0001-a
0003-a
0003-b
0003-c
0003-d
0015
0027-c
0035
0048
0050-a
0053
0054
0055
0060
0060-a
155
-------
diatoms (continued)
0067
0095
0097
0118
0126-d
0129
0148
Pi ctyosphaeri um
0139
Pictyosphaerium pulIchelTurn
0084
dimethyl sulfoxide
0118
diversity indices
0082
DMSO
0118
DMA
0006
DO
0050
dredging
0098
duckweed
0027-a
Dunaliella tertiolecta
0097
0098
dyes
0013-a
0065-a
0144-a
0147
ecological factors
0022
electronic particle counter
0117
0143
Euglena
0028
Euglena gracilis
0014
0017
0027-f
0029
0030
0030-b
0032
0087
Euglena gracilis Klebs
0028-a
eutrophication
0010
0011
0022-a
0025
0033
0046
0052
0059-a
0065
0066-a
0070
0074
0075
0076
0080-a
156
-------
eutrophication (continued)
fish
0085
0086-a
0087
0093
0095
0098-a
0102
0113
0113-a
0114
0115
0119
0120
0121
0125
0126
0126-a
0128
0128-a
0128-b
0128-d
0130
0132
0134
0135
0136
0137
0137-a
0140
0141
0146
0148
0149
experimental design
0003
0079
0108
0140
extraction
0027-d
feedlot runoff
0025
0142
fjord
0003-a
0003-b
0013-a
0037
0065-a
0129
0144-a
0054
0055
0119-a
fluorescence
0027-c
0117
0163
forecasting
0135
forest runoff
0066-a
fractionation
0109
0110
0125
Fremyella diplosiphon
0118
Gloeotrichia
0027-b
Gloeotrichia ecinulata
0084
glucose
0133
157
-------
Gomphonema
0050-a
growth inhibition
0067
growth kinetics
0007
0016-a
0060-a
0073
0086
0102
0126-d
0137-b
growth physiology
0047
growth rates
0003
0008-a
0009
0027-c
0040
0058
0074
0087
0093
0100
0101
0102
0102-a
0104-b
0105
0107
0114
0116
0117
0126-d
0127
0135
0139
0140
0141
0144
0147
growth rates (continued)
0151
0152
0154
0155
0157
0162
heavy metals
0013
0013-b
0037
0044
0044-a
0046
0049
0050
0060-a
0077
0078
0079
0080-a
0081
0095
0126
0144-a
0164
herbicides
0066
HMX
0003-b
hydrazine
0037
hydrocarbons
0089
hydrogen
0085
158
-------
impoundment
inhibition (continued)
0080-a
0113-a
0128-a
0128-b
0128-c
0128-d
0136
0137
0137-a
0137-b
0145
0146
0148
0149
0159
0161
0162
indigenous phytoplankton
0046
0074
indolebutyric acid
0050-a
inhibition
0013-b
0027-f
0043
0044
0048
0050-a
0065-a
0073
0078
0079
0080-a
0081
0086-a
0093
0095
0097
0104
0104-a
0104-b
0106
0107
0109
0110
0111
0123
0126
0140
0144-a
0164
inoculum
0154
inoculum age
0150-a
0151
0152
inorganic carbon
0086-a
0110
inorganics
0041
0058
insecticides
0001
interstitial waters
0098
iron
kinetics
0027-d
0034
0061
0084
0129
0008
0041
0087
159
-------
kinetics (continued)
lake waters (continued)
0099
0104-a
0104-b
0105
0134
0135
Kirchneriella lunaris
0004
Kirchneriella subsolitaria
0085-a
lake waters
0001-a
0008
0010
0011
0013
0014
0016
0017
0017-a
0018
0020-c
0022-a
0023
0026
0027
0027-a
0027-c
0027-d
0027-e
0031
0032
0033
0036
0038
0039
0044-a
0045
0046
0051
0055
0056
0059-a
0060-b
0061
0065
0066
0066-a
0069
0070
0071
0073
0074
0075
0076
0079
0080
0080-a
0090
0091
0093
0095
0098-a
0099
0102
0103
0108
0108-a
0112
0113-a
0114
0116
0119
0119-a
0120
0122
0126
0126-a
0126-b
0129
0130
0131
0132
0138
0150
0159
0160
0161
0162
0163
0166
leachates
0001-a
160
-------
lead limiting nutrients
(continued)
0013-b
0034 0053
0060-a 0054
0083 0055
0060-b
Lemna minor 0061
0068
0027-a 0069
0070
light 0073
0074
0027-c 0077
0047 0080
0085 0080-a
0094 0081
0140 0086
0147 0091
0094
light intensity 0100
0108
see light 0113
0114
light penetration 0115
0116
see light 0126
0126-a
limiting nutrients 0126~b
0127
0007 0128
0008-a 0128-c
0010 0131
0011 0134
0012 0136
0014 0137-a
0015 0137-b
0016 0138
0017 0140
0017-a 0143
0019 0145
0022 0146
0022-a 0150
0026 0161
0027-c 0162
0029-a 0166
0032
0033 lipids
0038
0042 0089
0046
0050
161
-------
luminescence
0006
MAAP
0082
manganese
0013-b
0019
0034
0084
mathematical model
0068
0090
0104-a
0106
0107
maximum yield
0027-c
media
0013-b
0073
0085-a
0108
0119-a
0126-c
0139
0150-a
0151
0152
0153
0154
0156
0157
0158
0159
0163
Melosira granulata
0148
membrane filtration
0025
mercury
0024
0027-b
0034
0050
mesotrophic
0025
metabolites
0165
Michaelis-Menten equation
0008
0086
0102
Microcystis
, 0027-a
0060-b
0148
Microcystis aeruginosa
0001
0003-a
0003-b
0003-c
0003-d
0006
0018
0020-b
0027
0027-b
0038
0048
0052
0057
0061
0082
0084
0087
0093
162
-------
Mlcrocystis aeruginosa
Navicula
0094
0095
0096
0097
0109
0123
0129
0137
0140
0148
Microcystls aeruginosa Kutzing
0145
micronutrients
0150-a
0151
0157
Mini test
0014
0016
0017
0028
0031
molybdenum
0041
0144
0150
morpholine
0037
mutagenicity
0165
mutual ism
0133
NAAM
0159
0050-a
Navicula pelliculosa
0003-a
0003-b
0003-c
0067
Navicula seminulum
0048
0095
0097
0129
nickel
nitrate
0013
0010
0028
0061
0124
0151
0154
0157
0166
nitrilotriacetate
0159
0160
nitrite
nitrogen
0028
0011
0012
0016-a
0017-a
0019
0020-b
0020-c
0022-a
163
-------
nitrogen (continued) nitrogen (continued)
0027 0128
0027-c 0128-a
0027-d 0128-b
0027-e 0128-c
0028-a 0128-d
0029-a 0129
0030-b 0130
0031 0131
0031-a 0132
0036 0134
0039 0136
0043 0137-b
0045 0140
0046 0145
0054 0146-a
0055 0148
0056 0157
0059-a 0158
0060-b 0159
0063 0160
0068 0162
0070 0164
0071 0166
0073
0075 nitrogen fixation
0076
0077 0052
0080 0146
0080-a
0081 nitrogen oxide
0085-a
0090 0075
0091
0098 nitroglycerine
0099
0105 0003-c
0108-a
0109 Nitzschia
0110
0112 0067
0113-a 0118
0114
0115 Nitzschia palea
0116
0120 0126-d
0124
0125 Nostoc
0126
0126-a 0007
0126-b
164
-------
Nostoc commune nutrients (continued)
0089 0027-a
0027-c
Nostac lino Id a 0027-e
0028-a
0067 0029
0030-b
Nostoc muscorum 0031-a
0050
0061 0052
0059-a
N:P ratios 0070
0073
0003-e 0074
0010 0075
0012 0076
0016 0079
0016-a 0085
0017-a 0085-a
0022-a 0094
0029-a 0098
0054 0101
0060-b 0102
0077 0103
0085-a 0104
0140 0108-a
0162 0111
0113-a
NTA 0114
0115
0082 0116
0129 0119-a
0159 0120
0160 0121
0122
nuclear power 0124
0125
0037 0126
0039 0126-a
0128-a
nutrients 0128-b
0128-c
0008-a 0128-d
0009 0130
0016-a 0131
0020 0132
0020-a 0135
0020-b 0136
0020-c 0137-b
0022-a 0138
0027 0140
165
-------
nutrients (continued)
0141
0142
0145
0146
0146-a
0148
0149
0152
0153
0156
0158
0160
0161
0162
0163
0166
oil
0104-a
0104-b
0106
oil shale
0001-a
Oocystis marssonii
0118
Oocystis submarina var. variabilis
0030
0030-b
organic fractions
0072
organics
0018
0088
0109
0110
0125
0133
0164
organochlorine compounds
0001
orthophosphate
0150
Oscillatoria
0027-b
Oscillatoria agardii
0015
Oscillatoria geminata
0038
Oscillatoria redekii
0091
Oscillatoria tenius
0038
Oscillatoria Vaucher
0119-a
oxygen
0049
0148
oxygen electrode
0050
oyster larvae
0165
32p
0008
166
-------
PAAP
phosphate (continued)
0082
0108
0140
0150-a
0151
0154
0155
0156
0157
0158
0163
PCB
0053
Pediastrum
0066
0026
0028
0030-b
0033
0037
0047
0056
0068
0083
0086
0096
0098-a
0113-a
0124
0129
0153
0154
0157
0164
0166
pH
phosphorus
0005
0041
0083
0086-a
0153
Phaeodactylum tricornutum
0015
0035
0053
0054
0055
0060
phenol
0104-b
0106
0107
phosphate
0007
0008
0008-a
0010
0023
0003-d
0007
0011
0016-a
0019
0020
0020-a
0020-c
0022-a
0026
0027
0027-a
0027-c
0027-d
0027-e
0027-f
0028
0028-a
0029-a
0030
0030-b
0031
0031-a
0033
0036
0038
0039
167
-------
phosphorus (continued) phosphorus (continued)
0043 0126-a
0044 0126-b
0045 0128-a
0046 0128-b
0051 0128-c
0053 0128-d
0054 0129
0055 0130
0057 0131
0059-a 0132
0060-b 0135
0061 0136
0062 0137-a
0063 0137-b
0066-a 0140
0070 0141
0071 0142
0072 0145
0073 0146
0074 0146-a
0075 0148
0076 0150
0077 0153
0080 0157
0080-a 0158
0081 0159
0084 0162
0085-a 0166
0086
0090 photohydrogen
0091
0094 0004
0095
0098 photosynthetic rates
0099
0100 0086-a
0101
0108-a phytoplankton
0109
OHO 0001-a
OUT 0003-b
0112 0006
0113 0008
0114 0019
0115 0022
0116 0026
0120 0027-a
0124 0027-c
0125 0027-d
0126 0027-e
168
-------
phytoplankton (continued)
pond water
0030
0034
0038
0045
0060-b
0066
0089
0091
0092
0095
0099
0112
0113
0119
0125
0126
0126-b
0131
0145
0148
0159
Pimephales promelas
0013-a
0065-a
0144-a
Poisson distribution
0003
pollution
0027-b
0027-e
0027-f
0032
0050
0052
0088
0104
0119
0119-a
0121
0127
0144-a
0164
0025
0162
Potomogeton
0148
precipitation
0153
pretreatment
0025
0051
0079
0111
0140
0143
0161
0162
protozoa
0001-b
Raphidiopsis curvata
0038
RDX
0003-a
refinery
0104-a
0106
reservoirs
0005
0022
0039
0046
0100
0101
0124
0125
0126
169
-------
reservoirs (continued) Salmo gairdneri
0141 0037
0142
salt loading
resolubilization
0001-a
0153
scenedesmine
restoration
0050-a
0065
Scenedesmus
Rhizoclonium
0004
0027-d 0028
0050-a
rivers 0083 .
0139
0002
0009 Scenedesmus obiiquus
0019
0020-a 0004
0020-b 0087
0020-c 0144
0027-d
0027-e Scenedesmus obtusiusculus
0034
0036 0083
0043
0044-a Scenedesmus quadricaruda
0045
0054 0001-a
0059 0014
0059-a 0017
0076 0027-f
0077 0029
0079 0030
0102 0030-b
0114 0032
0119 0041
0119-a 0058
0122 0060-a
0125 0067
0126-a 0089
0126-b 0117
0138 0118
0139 0127
0141
0142 Scenedesmus quadricauda
0149 Turpin em. Chod.
0161
0162 0028-a
170
-------
sediments
0026
0034
0074
0098
0125
Selenastrum
0004
0027-a
0028
0029-a
0050-a
0066
0124
0139
0150-a
0151
0152
0153
0154
0155
0156
Selenastrum 235 (UTEX)
0126-c
Selenastrum 747 (UTEX)
0126-c
Selenastrum bibraianum
0098-b
0098-c
Selenastrum bibraianum Reinsch 324 (UTEX)
0126-c
Selenastrum capricornutum
0001-a
0002
0003
0003-a
0003-b
0003-c
0003-d
Selenastrum capricornutum
(continued)
0005
0006
0007
0008
0009
0009-a
0010
0011
0012
0013
0013-a
0013-b
0014
0015
0016
0016-a
0017
0017-a
0018
0019
0020
0020-a
0020-b
0020-c
0022
0022-a
0024
0025
0026
0027
0027-b
0027-c
0027-d
0027-e
0027-f
0029
0030
0030-a
0030-b
0031
0031-a
0032
0033
0034
0035
0036
0037
0038
0039
0040
171
-------
Selenastrum capricornutum
(continued)
0041
0043
0044
0044-a
0045
0046
0048
0049
0050
0051
0052
0052-a
0053
0054
0055
0056
0057
0058
0060
0060-a
0060-b
0061
0062
0063
0064
0065
0065-a
0066-a
0067
0068
0069
0070
0072
0073
0074
0076
0077
0078
0080
0080-a
0081
0082
0083
0084
0085
0086
0086-a
0087
0090
0091
Selenastrum capricornutum
(continued)
0093
0094
0095
0096
0097
0098
0098-a
0099
0100
0101
0102
0103
0104
0104-a
0104-b
0105
0106
0107
0108
0108-a
0109
0110
0111
0113
0114
0117
0118
0119
0119-a
0120
0121
0123
0125
0126-a
0126-b
0126-d
0127
0128
0128-a
0128-c
0128-d
0129
0130
0131
0132
0133
0136
0137
0137-a
0138
172
-------
Selenastrum capricornutum
(continued)
0140
0141
0142
0143
0144-a
0145
0146
0147
0148
0149
0150
0157
0158
0159
0160
0161
0162
0163
0165
0166
Selenastrum capricornutum Printz
0001
0008-a
0023
0028-a
0047
0059
0059-a
0071
0079
0112
0113-a
0115
0116
0126
0128-b
0134
0135
0137-b
0146-a
0164
Selenastrum capricornutum
Printz CHL 1 (NIWR-)
0122
Selenastrum capricornutum
Printz 1648 (UTEX)
Selenastrum capricornutum
ATCC #22662
0001-b
Selenastrum gracile
0042
0075
0089
Selenastrum gracile Reinsch
0003-e
Selenastrum gracile
Reinsch B 325 (UTEX)
0126-c
Selenastrum gracile
(Strain 278-2)
0021
Selenastrum minutum
0102-a
Selenastrum minutum (Nag.)
Collins 326 (UTEX)
0126-c
Selenastrum westii
0085-a
0144
Selenastrum westii G.M.
Smith CHL 17 (NIWR)
0122
si 1ver
0027-b
Skeletonema costatum
0053
0054
0119-a
0126-c
173
-------
sodium chromate
0037
sodium citrate
0093
soil extraction
0003-e
sol utilization
0111
sorption
0026
0027-a
0027-d
species composition
0164
species diversity
0164
Sphaerocystis schroeteri
0045
Spirogyra
0027
0027-d
Spirulina platensis
0089
standing crop
0027-c
0090
0140
0145
statistical methods
0003
0160
0163
Steeman Nielson and v,
Brand technique
sterols
0126-d
0089
StigeocIonium tenue
0067
0130
stimulation
0001-a
0080-a
0093
0096
0098-a
0101
0104
0109
0110
0115
0126-a
0130
0133
0140
0142
0144-a
0159
0160
stratification
0125
streams
0020-c
0066-a
0069
0075
174
-------
succession
0145
sulfates
0037
sulfides
0049
surface:volume ratios
0155
0156
suspended solids
0135
Synedra delicatissina
0001-a
taconite
0098-a
tailings
0098-a
temperature
0040
0047
0104-a
0104-b
0105
0106
0107
0125
0140
Tetrae'dron bitridens
0118
1,3,5,7-tetranitro-octahydro-
1 ,3,5,7-tetrazocine
0003-b
TFM
time
TNT
toxicity
0067
0135
0123
0165
0003-a
0003-b
0003-c
0003-d
0009-a
0013
0013-a
0013-b
0017
0024
0027-b
0028
0030
0034
0037
0043
0044
0044-a
0048
0049
0050
0053
0065.-a
0074
0077
0079
0080-a
0081
0083
0087
0095
0096
0097
0098-a
0101
0104
0104-a
175
-------
toxicity (continued)
0104-b
0106
0107
0123
0126-a
0127
0129
0140
0144
0144-a
0148
0165
trace elements
0029-a
0064
0077
0078
0079
0085
0087
0095
0096
0097
0101
0122
0126-a
0140
0144
trinitrotoluene
0123
TRIS
0005
trisodium nitrilotriacetate
0082
0129
Ulothrix acuminata
0004
unsaponifiables
0089
uptake
0007
0008
0057
0086-a
0104-b
0105
0160
urban runoff
0020
0020-a
0066-a
Uronema gigas
0089
Uronema terrestre
0089
Volvox rousseletii
0145
wastewater
0009
0009-a
0013-b
0016
0017
0018
0023
0027-a
0027-d
0027-e
0027-f
0028
0029
0029-a
0030
0030-a
0030-b
0031
0031-a
0032
0033
0036
176
-------
wastewater (continued) wastewater (continued)
0042 0130
0043 0132
0044-a 0136
0045 0137-b
0053 0138
0054 0140
0055 0141
0056 0142
0063 0144-a
0065 0149
0069 0159
0070 0160
0072 0164
0074
0076 water hardness
0077
0079 0005
0080-a 0051
0081 0074
0091
0093 water quality
0094
0095 0003-a
0096 0003-b
0099 0003-c
0100 0009
0101 0027-a
0102 0027-e
0103 0029-a
0104 0031-a
0104-a 0034
0104-b 0044-a
0106 0045
0109 0060-b
0110 0069
0113 0076
0114 0080-a
0116 0114
0119 0120
0119-a 0121
0121 0126
0125 0127
0126 0130
0126-a 0142
0127 0143
0128 0161
0128-a
0128-c watershed
0128-d
0129 0124
177
-------
l-i
zinc
0108
0127
0002
0013
0034
0037
0044
0044-a
0045
0046
0049
0050
0060-a
0078
0080-a
0081
178
-------
3.0 Author Index
Adam, V.D.
0001-a
Adinarayana, A.
0075
American Type Culture Collection
0001-b
Appleman, R.
0113
Armstrong, D.E.
0062
Armstrong, R.
0042
Aronson, J.G.
0001
Ashton, P.J.
0145
Auer, M.T.
0057
Bartlett, L.
0002
0034
Behnken, D.W.
0003
Bennett, P.J.
0034
Bentley, R.E.
0003-a
0003-b
0003-c
0003-d
Bilcea, R.
0003-e
Bishop, N.I.
0004
Blind, N.L.
0070
Bliss, J.D.
0060-b
Bowman, R.H.
0005
Brezonik, P.L.
0006
Brown, E.J.
0007
0008
0008-a
Brown, J.W.
0051
Browne, F.X.
0006
Bruwer, M.J.
0137
0137-a
179
-------
Button, O.K.
0008-a
Buxton, K.S.
0003-c
Cain, J.R.
0009
0059
Cairn, J., Jr.
0166
Camp, F.A.
0009-a
0166
Capella, P.
0089
Chiaudani, G.
0010
0011
0012
0013
Chillingsworth, M.B.
0013-a
0065-a
Christensen, E.R.
0013-b
Claesson, A.
0014
0015
0016
0016-a
0017
0017-a
0029-a
0031
Claesson, A. (continued)
0031-a
0032
0108-a
Clark, J.R.
0123
Cleave, M.L.
0001-a
Clesceri, N.L.
0018
0023
0072
0073'
0110
Coleridge, S.E.
0051
Condit, R.J.
0019
0034
Connell, A.D.
0113-a
Cooke, G.D.
0066-a
Cowan, P.A.
0100
0101
Cowen, W.F.
0020
0020-a
0020-b
0020-c
180
-------
Czygan, F.C.
0021
Dean, J.W.
0003-a
0003-c
0003-d
Denison, J.R.
0022
DiSalvo, L.H.
0165
Dixon, P.S.
0052-a
0085
0113
Dolan, J.M.
0009-a
Dunigan, P.F.X.
0034
Dye, C.
0022-a
Ells, S.J.
0003-a
0003-c
Falkenborg, D.H.
0074
Faust, S.L.
0027-d
Ferris, J.J.
0023
FIT by, R.
0034
Filip, D.S.
0024
0025
Fitzgerald, G.P.
0026
0027
0027-a
0027-b
0027-c
0027-d
0027-e
Florenzano, G.
0089
Forsberg, A.
0027-f
0028
0028-a
0030
0031-a
0032
0108-a
Forsberg, C.G.
0029
0029-a
0030
0030-a
0030-b
0031
0031-a
0032
Fox, J.L.
0006
Francisco, D.E.
0033
181
-------
Frick, M.
0004
Funk, W.H.
0002
0034
Gaonkar, S.A.
0087
Gargas, E.
0035
0036
Garton, R.B.
0037
Gasperino, A.F.
0046
0125
0126
Gavrila, L.
0050-a
Geissel, L.D.
0067
Gerhold, R.M.
0038
0039
Gerloff, G.C.
0027-c
Goldman, C.R.
0042
Goldman, J.C.
0040
0041
Grau, P.
0102
Green, W.J.
0018
Greene, J.C.
0043
0044
0044-a
0045
0046
0077
0078
0079
0080
0080-a
0081
0115
0116
Grenney, W.J.
0068
0104-b
0105
0106
0107
Griffin, J.P.
0126
Haaland, P.T.
0047
Hall, R.H.
0048
0096
0097
Hand, J.
0022-a
182
-------
Harris, R.F.
0007
0008
0062
Hartwell, K.
0131
Helms, R.W.
0163
Hendricks, A.C.
0009-a
0049
Hern, S.C.
0060-b
Hokerval1, E.
0030-a
0030-b
Hollister, T.A.
0003-a
0003-b
0003-d
Hostetter, H.P.
0050
Hoyle, J.L.
0114
Huang, C.H.
0102
0135
0138
Hyman, K.L.
lonescu, A.
0050-a
Jadlocki, J.F. , Jr.
0051
0111
James, N.G.
0165
Jenkins, D.
0041
Johnson, H.E.
0067
Joint Industry/Government Task
Force on Eutrophication
0052
Jones. D.
0022-a
Jones, L.W.
0004
0137
0137-a
Justice, C.
0052-a
0113
Kallqvist, T.
0053
0054
0055
Katko, A.
0056
Keenan, J.D.
0057
183
-------
King, D.L.
0058
Klotz, R.L.
0059
Knutson, G.
0047
Kobayashi, S.
0023
Kohlmeyer, S.I.
0113-a
0137-b
Koonce, J.K.
0008
Kotai, J.
0059-a
0060
Krogh, T.
0059-a
0060
Kruger, D.M.
0126
Kumar, I.S.
0018
Laake, M.
0060-a
Lambou, V.W.
0060-b
Lange, W.
0061
Lavin, P.J.
0073
LeBlanc, G.A.
0003-a
0003-b
0003-c
0003-d
Lee, C.C..
Lee, G.F.
0062
0020
0020-a
0020-b
0020-c
0098-a
0126-a
0126-b
0150
Lee, T.H.
0084
Lehmusluoto, P.O.
0063
Lepers, A.
0102-a
Lercker, G.
0089
Lindemann, E.G.
0064
Lindmark, G.
0065
184
-------
Lingg, A.J.
0133
Little, L.
0066
Little, L.W.
0065-a
Lium, B.W.
0117
0118
Long, E.T.
0066-a
Lynn, R.I.
0024
Maki, A.W.
0067
Mai one, R.F.
0068
Maloney, I.E.
0043
0069
0070
0071
0074
0076
0080
McDonald, G.C.
0018
0072
0073
McGauhey, P.H.
0075
Merwin, E.A.
0044
0080-a
0081
Middlebrooks, E.J
0025
0074
0075
0100
0101
0104-a
0104-b
0105
0106
0107
Miller, W.E.
0043
0044
0044-a
0045
0046
0069
0070
0071
0076
0077
0078
0079
0080
0080-a
0081
0115
0116
0131
Mitchell, D.
0082
Monahan, T.J.
0083
Morton, S.D.
0084
185
-------
Murray, S.
0052-a
0085
Nadler, C.R.
0027-d
Natarajan, K.V.
0085-a
0144
Nicholson, C.S.
0051
Novak, J.T.
0058
Nyholm, N.
0086
Olofsson, J.A., Jr.
0086-a
Ormerod, K.
0060
Oswald, W.J.
0041
0087
Otta, R.G.
0039
Palmer, M.
0088
Paoletti, C.
0089
Parker, J.I.
0034
Parker, M.
0090
Parr, M.P.
0091
Parra, B.O.
0092
Payne, A.G.
0093
0094
0095
0096
0097
0129
Pearson, E.A.
0075
0102
0138
Pittman, D.
0066
Plumb, R.H., Jr.
0098
0098-a
Polesco-Ionasesco, L.
0098-b
Polescu, L.
0098-c
Porcella, D.B.
0001-a
0099
186
-------
Porcella, D.B. (continued)
0100
0101
0102
0104-a
0104-b
0105
0106
0107
Premazzi, G.
0102-a
Proctor and Gamble
0103
Pushparaj, G.
0089
Putnam, K.
0044-a
0045
Rabe, F.W.
0002
0034
Radimsky, J.
0102
0138
Ravera, 0.
0102-a
Raw!ings, G.D.
0104
Reynolds, J.H.
0068
0104-a
0104-b
0105
Reynolds, J.H. (continued)
0106
0107
Rodhe, W.
0108
Rolich, G.A.
0075
Ross, L.
0022-a
Rothman, E.D.
0112
Ryding, S.-O.
0017-a
0031
0031-a
0032
0108-a
Sachdev, D.R.
0109
0110
Saldick, J.
0051
0111
Sauter, S.
0003-a
0003-c
0003-d
Savage, N.L.
0034
187
-------
Scherfig, J.
Scott, W.E.
Seaman, M.T.
Shaw, K.
Sherrarc
Shiroyama, T.
Shoaf, W.T.
J.
0013-b
0052-a
0085
0113
0138
C.L.
0112
0113-a
0128-a
0147
T.
0113-a
0034
J.H.
0114
T.
0043
0044
0044-a
0045
0046
0071
0077
0078
0079
0080-a
0081
0115
0116
0117
0118
Shubert, I.E.
0139
Simmons, M.S.
0112
Sirisinha, K.
0020-b
0020-c
Skulberg, O.M.
0059-a
0060
0119
0119-a
0120
0121
0122
Sleight, B.H.
0003-a
0003-b
0003-c
0003-d
Smith, R. L.
0132
Smith, R.V.
0091
Smith, W.W.
0051
Smock, L.A.
0123
Smolen, M.D.
0124
188
-------
Soltero, R.A.
0044-a
0045
0046
0125
0126
Spear, R.D.
0073
Sridharan, N.
0126-a
0126-b
Starr, R.C.
0126-c
Steeman Nielson, E.
0126-d
Steensland, H.
0127
Steyn, D.J.
0128
0128-a
0128-b
0128-c
0128-d
0136
0137-b
0148
0149
Stoneburner, D.L.
0123
Sturm, R.N.
0129
Sugiki, A.
0130
Syers, J.K.
0062
Thomas, N.A.
0131
Thomas, R.E.
0132
Thomas, R.W.
0060-b
Thomas, S.R.
0125
0126
Thompson, N.
0034
Tison, D.L.
0133
Toerien, D.F.
0102
0113-a
0128
0128-a
0128-b
0128-c
0128-d
0134
0135
0136
0137
0137-a
0137-b
0138
0146
0146-a
0147
0148
0149
189
-------
Torrey, M.S.
0027-e
Trainor, F.R.
0009
0059
0139
Tunzi, M.GV
0042
0075
U.S. Environmental Protection Agency
0140
0141
0142
U.S. Geological Survey
0143
Venkataraman, G.S.
0144
Vighi, M.
0010
0011
0012
0013
Visser, J.H.
0128
0128-a
0128-b
0128-c
0128-d
Volpp, G.P.
0064
Voos, K.A.
0068
Vyas, H.
0144-a
Walmsley, R.D.
0145
0146
0146-a
0147
0148
0149
Walton, C.P.
0150
Watts, D.G.
0003
Weiss, C.M.
0033
0150-a
0151
0152
0153
0154
0155
0156
0157
0158
0159
0160
0161
0162
0163
Williams, L.R.
0060-b
Williams, P.M.
0125
0126
0164
Wilson, W.G.
0003-d
190
-------
Winner, J.E.
0034
Won, W.D.
0165
Woodard, F.E.
0086-a
Wright, J. , Jr.
0166
191
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
REPORT NO.
EPA-600/9-79-021
4. TITLE AND SUBTITLE
2.
M I i_ e MINUSUCII i i_ t
Bibliography of Literature Pertaining to the Genus
Sf>l pna^l-rnm
Selenastrum
3. RECIPIENT'S ACCESSION NO.
5. REPORT DATE
July 1979 issuing date
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Amy A. Leischman, Joseph C. Greene, and
William E. Miller
8. PERFORMING ORGANIZATION REPORT NO.
9 PERFORMING nor;Awi-7A-rinw NAMF AND ADDRESS
Environmental Research Laboratory--Corval1is
Office of Research and Development
U.S. Environmental Protection Agency
Corvallis, Oregon 97330
10. PROGRAM ELEMENT NO.
IBA608
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
Same as above
13. TYPE OF REPORT AND PERIOD COVERED
Final
14. SPONSORING AGENCY CODE
EPA/600/02
15. SUPPLEMENTARY NOTES
This report supplements the Selenastrum capricornutum Printz Algal Assay
Test EPA 600/9-78-018
Bottle
16. ABSTRACT
This bibliography supplements the 1978 Algal Assay protocol by compiling the
aveilable literature, published and unpublished, on research that contained
algal assays using Selenastrum. The papers cited in this bibliography include
studys of the effects of nutrients, toxicants,, complex wastes, and specific
inorganic and organic compounds upon Selenastrum's growth and physiology. The
result is a collection of abstracts, citations, and key words describing 220
papers. This bibliography is designed to be used as a public reference for
managers of water quality programs and scientists or educators with interests
in research pertaining to Selenastrum and algal assays.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS
c. COSATI Field/Group
Bibliography, Algal assay, Selenastrum,
nutrients, toxicity, wastewater,
herbicides, author index, key work index
08/H
8. DISTRIBUTION STATEMEN1
Unlimi ted
19. SECURITY CLASS (This Report)
Unclassi fled
21. NO. OF PAGES
199
20. SECURITY CLASS (This page)
Unclass if led
22. PRICE
EPA Form 2220-1 (Rev. 4-77) PREVIOUS EDITION is OBSOLETE
192
•6GPO 699-322
------- |