&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

-------
                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.

-------
                                                 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

-------
                                  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.

-------
                                   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

-------
                                    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

-------
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.

-------
     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.

-------
                               TABLE OF CONTENTS



                                                                          Page



Foreword	   iii



Preface	    iv



Bibliography 	     1



Keyword Index  	   144



Author Index 	   179
                                     VI1

-------
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.

-------
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

-------
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.

-------
     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).

-------
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).

-------
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

-------
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

-------
     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

-------
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

-------
  -  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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
     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

-------
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

-------
     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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
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

-------
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

-------
     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

-------
     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

-------
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

-------
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

-------
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

-------
     (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

-------
     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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
     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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
     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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
     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

-------
     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

-------
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

-------
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

-------
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

-------
     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

-------
     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

-------
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

-------