EPA-600/3-77-124
October 1977
Ecological Research Series
            DISTRIBUTION  OF  PHYTOPLANKTON IN
                                     MARYLAND  LAKES
                                Environmental Monitoring and Support Laboratory
                                        Office of Research and Development
                                       U.S. Environmental Protection Agency
                                              Las Vegas. Nevada 89114

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                RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination  of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:

      1.   Environmental Health Effects Research
      2.   Environmental Protection Technology
      3.   Ecological Research
      4.   Environmental Monitoring
      5.   Socioeconomic Environmental Studies
      6.   Scientific  and Technical Assessment Reports (STAR)
      7.   Interagency Energy-Environment Research and Development
      8.   "Special" Reports
      9.   Miscellaneous Reports

This report has been assigned to the ECOLOGICAL RESEARCH series. This series
describes research on the effects of pollution on humans, plant and animal spe-
cies, and materials. Problems are assessed for their long- and short-term influ-
ences. Investigations include formation, transport, and pathway studies to deter-
mine the fate of pollutants and their effects. This work provides the technical basis
for setting standards to minimize undesirable changes in living organisms in the
aquatic, terrestrial, and atmospheric  environments.
 This document is available to the public through the National Technical Informa-
 tion Service, Springfield, Virginia 22161.

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                                                EPA-600/3-77-124
                                                October 1977
DISTRIBUTION OF PHYTOPLANKTON IN MARYLAND LAKES


                        by
V. W. Lambou, F. A.  Morris*, R.  W.  Thomas, M.  K.  Morris*,
       L.  R.  Williams, W.  D. Taylor,  F.  A. Hiatt*,
               S.  C. Hern, and J.  W.  Hilgert*
         Monitoring Operations Division
 Environmental  Monitoring and Support Laboratory
            Las Vegas, Nevada   89114
       *Department of Biological  Sciences
       The University of Nevada,  Las Vegas
            Las Vegas, Nevada   89154
   ENVIRONMENTAL MONITORING AND SUPPORT LABORATORY
         OFFICE OF RESEARCH AND DEVELOPMENT
        U.S. ENVIRONMENTAL PROTECTION AGENCY
              LAS VEGAS, NEVADA    89114

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                              DISCLAIMER
     This report has been reviewed by the Environmental Monitoring and
Support Laboratory-Las Vegas, U.S. Environmental Protection Agency,
and approved for publication.  Mention of trade names or commercial
products does not constitute endorsement or recommendation for use.
                                    1.1

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                                 FOREWORD
     Protection of the environment requires effective regulatory
actions which are based on sound technical  and scientific information.
This information must include the quantitative description and linking
of pollutant sources, transport mechanisms, interactions, and re-
sulting effects on man and his environment.  Because of the complexities
involved, assessment of specific pollutants in the environment requires
a total systems approach which transcends the media of air, water,  and
land.  The Environmental Monitoring and Support Laboratory-Las Vegas
contributes to the formation and enhancement of a sound integrated  moni-
toring data base through multidisciplinary, multimedia programs designed
to:

          •  develop and optimize systems and strategies for moni-
             toring pollutants and their impact on the environment

          •  demonstrate new monitoring systems and technologies
             by applying them to fulfill special  monitoring needs
             of the Agency's operating programs

     This report presents the species and abundance of phytoplankton
in the 17 lakes sampled by the National Eutrophication Survey in the
State of Maryland, along with results from the calculation of several
commonly used biological indices of water quality and community structure.
These data can be used to biologically characterize the study lakes,
and as baseline data for future investigations.  This report was written
for use by Federal, State, and local governmental agencies concerned with
water quality analysis, monitoring, and/or regulation.  Private industry
and individuals similarly involved with the biological aspects of water
quality will find the document useful.  For further information contact
the Water and Land Quality Branch, Monitoring Operations Division.
                                   George B.  Morgan
                                       Director
                 Environmental  Monitoring and Support Laboratory
                                     Las  Vegas

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                               CONTENTS

Foreword 	i i i
Introduction 	  1
Materials and Methods 	  2
     Lake and Site Selection 	  2
     Sample Preparation 	  2
     Examination 	  3
     Quality Control 	  4
Results 	  5
     Nygaard's Trophic State Indices 	  5
     Palmer's Organic Pollution Indices 	  5
     Species Diversity and Abundance Indices 	  7
     Species Occurrence and Abundance 	  9
Literature Cited 	 10
Appendix.  Summary of Phytoplankton Data 	 11

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                             INTRODUCTION
     The collection and analysis of phytoplankton data were included
in the National  Eutrophication Survey in an effort to determine
relationships between algal  characteristics and trophic status  of
individual lakes.

     During spring, summer,  and fall  of 1973,  the Survey sampled 250
lakes in 17 States.  Over 700 algal  species and varieties were
identified and enumerated from the 743 water samples  examined.

     This report presents the species and abundance of phytoplankton
in the 4 lakes sampled in the State of Maryland (Table 1).   The
Nygaard's Trophic State (Nygaard 1949), Palmer's Organic Pollution
(Palmer 1969), and species diversity and abundance indices  are  also
included.
             TABLE 1.  LAKES SAMPLED IN THE STATE  OF MARYLAND


STORET No.            Lake Name                     County


2402                 Deep Creek Lake               Garrett

2403                 Liberty Reservoir             Carroll,  Baltimore

2408                 Loch Raven Reservoir          Baltimore

2409                 Johnson Pond                  Wicomico

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                        MATERIALS AND METHODS
LAKE AND SITE SELECTION

     Lakes and reservoirs included in the Survey were selected through
discussions with State water pollution agency personnel  and U.S.
Environmental Protection Agency Regional Offices (U.S. Environmental
Protection Agency 1975).  Screening and selection strongly emphasized
lakes with actual or potential accelerated eutrophication problems.
As a result, the selection was limited to lakes:

     (1)  impacted by one or more municipal sewage treatment plant
          outfalls either directly into the lake or by discharge to  an
          inlet tributary within approximately 40 kilometers of the
          lake;

     (2)  40 hectares or larger in size; and

     (3)  with a mean hydraulic retention time of at least 30 days.

Specific selection criteria were waived for some lakes of particular
State interest.

     Sampling sites for a lake were selected based on available
information on lake morphometry, potential major sources of nutrient
input, and on-site judgment of the field limnologist (U.S.
Environmental Protection Agency 1975).  Primary sampling sites were
chosen to reflect the deepest portion of each major basin in a test
lake.  Where many basins were present, selection was guided by
nutrient source information on hand.  At each sampling site, a depth-
integrated phytoplankton sample was taken.  Depth-integrated samples
were uniform mixtures of water from the surface to a depth of 15 feet
(4.6 meters) or from the surface to the lower limit of the photic zone
representing 1 percent of the incident light, whichever was greater.
If the depth at the sampling site was less than 15 feet (4.6 meters),
the sample was taken from just off the bottom to the surface.
Normally, a lake was sampled three times in 1 year, providing
information on spring, summer, and fall conditions.
SAMPLE PREPARATION

     Four milliliters  (ml) of Acid-Lugo!'s sollution (Prescott 1970)
were added to each 130-ml sample from each site at the time of
collection for  preservation.  The samples were shipped to the
Environmental Monitoring and Support Laboratory, Las Vegas, Nevada,
where equal volumes from each site were mixed to form two 130-ml
composite samples for  a given lake.  One composite sample was put into
storage and the other  was used for the examination.

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     Prior to examination, the composite samples were concentrated by
the settling method.  Solids were allowed to settle for at least 24
hours prior to siphoning off the supernate.  The volume of the
removed supernate and the volume of the remaining concentrate were
measured and concentrations determined.  A small (8 ml) library
subsample of the concentrate was then taken.  The remaining
concentrate was gently agitated to resuspend the plankton and poured
into a capped, graduated test tube.  If a preliminary examination of a
sample indicated the need for a more concentrated sample, the contents
of the test tube were further concentrated by repeating the settling
method.  Final concentrations varied from 15 to 40 times the original.

     Permanent slides were prepared from concentrated samples after
analysis was complete.  A drop of superconcentrate from the bottom of
the test tube was placed in a ring of clear Karo Corn Syrup with
phenol (a few crystals of phenol were added to each 100 ml  of syrup)
on a glass slide, thoroughly mixed, and topped with a coverglass.
After the syrup at the edges of the coverglass had hardened, the
excess was scraped away and the mount was sealed with clear fingernail
polish.  Permanent diatom slides were prepared by drying sample
material on a coverglass, heating in a muffle furnace at 400  C for 45
minutes, and mounting in Hyrax .  Finally, the mounts were  sealed with
clear fingernail polish.

     Backup samples, library samples, permanent sample slides, and
Hyrax-mounted diatom slides are being stored and maintained at the
U.S.  Environmental Protection Agency's Environmental Monitoring and
Support Laboratory-Las Vegas.
EXAMINATION

     The phytoplankton samples were examined with the aid of binocular
compound microscopes.  A preliminary examination was performed to
precisely identify and list all forms encountered.   The length of this
examination varied depending on the complexity of the sample.   An
attempt was made to find and identify all  of the forms present in each
sample.  Often forms were observed which could not be identified to
species or to genus.  Abbreviated descriptions were used to keep a
record of these forms (e.g., lunate cell,  blue-green filament,
Navicula #1).  Diatom slides were examined using a standard light
microscope.  If greater resolution was essential to accurately
identify the diatoms, a phase-contrast microscope was used.

     After the species list was compiled,  phytoplankton were enumerated
using a Neubauer Counting Chamber with a 40X objective lens and a 10X
ocular lens.  All forms within each field were counted.  The count was
continued until a minimum of 100 fields had been viewed, or until the
dominant form had been observed a minimum of 100 times.
^Registered Trademark

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QUALITY CONTROL

     Internal quality control checks on species identifications and
counts were performed on a regular basis between project phycologists
at the rate of 7 percent.  Although an individual had primary
responsibility for analyzing a sample, taxonomic problems were
discussed among the phycologists.

     Additional quality control checks were performed on the Survey
samples by Dr. G. W. Prescott of the University of Montana at the rate
of 5 percent.  Quality control checks were made on 75 percent of these
samples to verify species identifications while checks were made on
the remaining 25 percent of  the samples to verify genus counts.
Presently, the agreement between quality control checks for species
identification and genus enumerations is satisfactory.

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                               RESULTS
     The Appendix summarizes all  of the phytoplankton data collected
from the State by the Survey.   It is organized by lake, including an
alphabetical phytoplankton species list with concentrations for
individual species given by sampling date.   Results from the
application of several indices are presented (Nygaard's Trophic State,
Palmer's Organic Pollution, and species diversity and abundance).
Each lake has been assigned a four digit STORET number.  [STORET
(STOrage and RETrieval) is the U.S. Environmental Protection Agency's
computer system which processes and maintains water quality data.]  The
first two digits of the STORET number identify the State; the last two
digits identify the lake.


NYGAARD'S TROPHIC STATE INDICES

     Five indices devised by Nygaard (1949) were proposed under the
assumption that certain algal  groups are indicative of levels of
nutrient enrichment.  These indices were calculated in order to aid in
determining the surveyed lakes' trophic status.  As a general rule,
Cyanophyta, Euglenophyta, centric diatoms,  and members of the
Chlorococcales are found in waters that are eutrophic (rich in
nutrients), while desmids and many pennate  diatoms generally cannot
tolerate high nutrient levels and so are found in oligotrophic waters
(poor in nutrients).

     In applying the indices to the Survey  data, the number of taxa in
each major group was determined from the species list for each sample.
The ratios of these groups give numerical values which can be used as
a biological index of water richness.  The  five indices and the ranges
of values established for Danish lakes by Nygaard for each trophic
state are presented in Table 2.  The appropriate symbol, (E) eutrophic
and (0) oligotrophic, follows each calculated value in the tables in
the Appendix.  A question mark (?) was entered in these tables when
the calculated value was within the range of both classifications.


PALMER'S ORGANIC POLLUTION INDICES

     Palmer (1969) analyzed reports from 165 authors and developed
algal pollution indices for use in rating water samples with high
organic' pollution.  Two lists of organic pollution-tolerant forms were
prepared, one containing 20 genera, the other, 20 species (Tables 3
and 4).  Each form was assigned a pollution index number ranging from
1  for moderately tolerant forms to 6 for extremely tolerant forms.
Palmer based the index numbers on occurrence records and/or where
emphasized by the authors as being especially tolerant of organic
pollution.

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 TABLE  2.   NYGAARD'S TROPHIC STATE INDICES ADAPTED FROM HUTCHINSON (1967)
  Index
    Calculation
Oligotrophic   Eutrophic
  Myxophycean


  Chlorophycean


  Diatom


  Euglenophyte


  Compound
     Myxophyceae
      Desmideae

     Chlorococcales
       Desmideae

     Centric Diatoms
     Pennate Diatoms

      Euglenophyta
Myxophyceae + Chlorococcales

Myxophyceae + Chlorococcales +
Centric Diatoms + Euglenophyta
          Desmideae
 0.0-0.4
 0.0-0.7
 0.0-0.3
 0.0-0.2
 0.0-1.0
0.1-3.0
0.2-9.0
0.0-1.75
0.0-1.0
1.2-25
TABLE 3.  ALGAL GENUS POLLUTION INDEX
          (Palmer 1969)
                       TABLE 4.  ALGAL SPECIES POLLUTION
                                 INDEX (Palmer 1969)
Genus
Anacystis
Ankistrodesmus
Chlamydomonas
Chlorella
Closterium
Cyclotella
Euglena
Comphonema
Lepocinclis
Melosira
Micractiniwn
ffavicula
Nitzschia
Oscillatoria
Pandorina
Phacus
Phormidium
Scenedesmus
Stigeoelonvum
Synedra
Pollution
Index
1
2
4
3
1
1
5
1
1
1
1
3
3
5
1
2
1
4
2
2
                                        Species
                                                 Pollution
                                                   Index
                                         Ankistrodesmus fatcatus      3
                                         Arthrospira jenneri          2
                                         Chlorella vulgaris           2
                                         Cyclotella meneghiniana      2
                                         Euglena gracilis             1
                                         Euglena viridis              6
                                         Gomphonema parvulum          1
                                         Melosira varians             2
                                         Navicula aryptocephala       1
                                         Nitzschia acicularis         1
                                         Nitzschiapalea              5
                                         Oscillatoria chlorina        2
                                         Oscillatoria linaysa          4
                                         Oscillatoria princeps        1
                                         Oscillatoria putrida         1
                                         Oscillatofia tenuis          4
                                         Pandorina rnorwn      .        3
                                         Scenedesmus quddricauda      4
                                         Stigeoclonium tenue          3
                                         Simedra ulna	3

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    In analyzing a water sample, any of the 20 genera or species of
algae present in concentrations of 50 per milliliter or more are
recorded.  The pollution index numbers of the algae present are
totaled, providing a genus score and a species score.  Palmer
determined that a score of 20 or more for either index can be taken as
evidence of high organic pollution, while a score of 15 to 19 is taken
as probable evidence of high organic pollution.   Lower figures suggest
that the organic pollution of the sample is not  high, that the sample
is not representative, or that some substance or factor interfering
with algal persistence is present and active.
SPECIES DIVERSITY AND ABUNDANCE INDICES

    "Information content" of biological samples is  being used
commonly by biologists as a measure of diversity.   Diversity in  this
connection means the degree of uncertainty attached to the specific
identity of any randomly selected individual.   The  greater the number
of taxa and the more equal their proportions,  the greater the
uncertainty, and hence, the diversity (Pielou  1966).   There are
several methods of measuring diversity, e.g.,  the formulas given by
Brillouin (1962) and Shannon and Weaver (1963).  The method which is
appropriate depends on the type of biological  sample on hand.

    Pielou (1966) classifies the types of biological  samples and
gives the measure of diversity appropriate for each type.   The Survey
phytoplankton samples are what she classifies  as larger samples
(collections in Pielou's terminology) from which random subsamples  can
be drawn.  According to Pielou, the average diversity per individual
for these types of samples can be estimated from the Shannon-Wiener
formula (Shannon and Weaver 1963):
                        S
                       •I
                       1 = 1
H  =  -I  PI  logx PI.
Where P is the proportion of the ith  taxon  in  the  sample, which  is cal-
culated from n./N; n-  is the number of individuals  per milliliter of the
ith taxon; N ii the total number of individuals  per ml;  and S  is the total
number of taxa.

    However, Basharin  (1959) and Pielou (1966) have pointed out  that
H calculated from the  subsample is  a  biased estimator of the sample H,
aad if this bias is to be accounted for,  we must know the total  num-
ber of taxa present in the sample since the magnitude of this  bias
depends on it.

    Pielou (1966) suggests that if  the number  of taxa in the
subsample falls only slightly short of the  number  in the larger
sample, no appreciable error will result in considering  S, estimated

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from the subsample, as being equal to the sample value.   Even though
considerable effort was made to find and identify all taxa, the Survey
samples undoubtedly contain a fair number of rare phytoplankton taxa
which were not encountered.

    In the Shannon-Wiener formula, an increase in the number of taxa
and/or an increase in the evenness of the distribution of individuals
among taxa will increase the average diversity per individual from its
minimal value of zero.  Sager and Hasler (1969) found that the
richness of taxa was of minor importance in determination of average
diversity per individual for phytoplankton and they concluded that
phytoplankton taxa in excess of the 10 to 15 most abundant ones have
little effect on H, which was verified by our own calculations.  Our
counts are in number per milliliter and since logarithms to the base
2 were used in our calculations, H is expressed in units of bits per
individual.  When individuals of a taxon were so rare'that they were
not counted, a value of 1/130 per milliliter or 0.008 per milliliter
was used in the calculations since at least one individual of the
taxon must have been present in the collection.

    A Survey sample for a given lake represents a composite of all
phytoplankton collected at different sampling sites on a lake during a
given sampling period.  Since the number of samples (M)  making up a
composite is a function of both the complexity of the lake sampled and
its size, it should affect the richness-of-taxa component of the
diversity of our phytoplankton collections.  The maximum diversity
(MaxH) (i.e., when the individuals are distributed among the taxa as
evenly as possible) was estimated from Iog2 S, the total diversity (D)
was calculated from HN, and the evenness component of diversity (J)
was estimated from H/MaxH  (Pielou 1966).  Also given in the Appendix
are L  (the mean number of individuals per taxa per milliliter) and K
(the number of individuals per milliliter of the most abundant taxon
in the sample).

    Zand (1976) suggests that diversity indices be expressed in units
of "sits", i.e., in logarithms to base S (where S is the total number
of taxa in the sample) instead of in "bits", i.e., in logarithms to
base 2.  Zand points out that the diversity index in sits per
individual is a normalized number ranging from 1 for the most evenly
distributed samples to 0 for the  least evenly distributed samples.
Also,  it can be used to compare different samples, independent of the
number of taxa in each.  The diversity in bits per individual should
not be used in direct comparisons involving various samples which
have different numbers of species.  Since MaxH equals log S, the ex-
pression in sits is equal to logs S, or 1.  Therefore diversity in sits
per individual is numerically equivalent to J, the evenness component
for the Shannon-Wiener formula.
                                   8

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SPECIES OCCURRENCE AND ABUNDANCE

    The alphabetic phytoplankton species  list for each  lake,
presented in the Appendix,  gives the concentrations  of  individual
species by sampling date.   Concentrations  are in  cells,  colonies,  or
filaments (CEL, COL, FIL)  per milliliter.   An "X" after a  species  name
indicates the presence of  the species on  that date in such a  low
concentration that it did  not show ,up in  the  count.  A  blank  space
indicates that the organism was  not  found  in  the  sample  collected  on
that date.  Column S is used to  designate  the examiner's subjective
opinion of the five dominant taxa in a sample,  based upon  relative
size and concentration of  the organism.  The  percent column (%C)
presents, by abundance, the percentage composition of each taxon.

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                           LITERATURE CITED
Basharin, G. P.  1959.  On a statistical estimate for the entrophy of a
    sequence of independent random variables, pp. 333-336.  In N.
    Artin (ed.), Theory of Probability and Its Applications
    (translation of "Teoriya Veroyatnosei i ee Premeneniya") 4.
    Society for Industrial and Applied Mathematics, Philadelphia.

Brillouin, L.  1962.  Science and Information Theory (2nd ed.).
    Academic Press, New York.  351 pp.

Hutchinson, G. E.  1967.  A Treatise on Limnology.  II.   Introduction
    to Lake Biology and the Limnoplankton.  John Wiley and Sons,
    Inc., New York.  1,115 pp.

Nygaard, G.  1949.  Hydrobiological studies of some Danish ponds and
    lakes.  II.  (K danske Vidensk.  Selsk.)  Biol.  Sci. 7:293.

Palmer, C. M.  1969.  A composite rating of algae tolerating organic
    pollution.  J. Phycol.  5:78-82.

Pielou, E. C.  1966.  The measurement of diversity in different types
    of biological collections.  J. Theor. Biol.  13:131-144.

Prescott, G. W.  1970.  How to Know the Freshwater Algae.  William C.
    Brown Company, Dubuque.  348 pp.

Sager, P. E. and A. D. Hasler.  1969.  Species diversity in laucustrine
    phytoplankton.  I.  The components of the index of diversity from
    Shannon's formula.  Amer. Natur.  103(929):51-59.

Shannon, C. E. and W. Weaver.  1963.  The Mathematical Theory of
    Communication.  University of Illinois Press, Urbana.  117 pp.

U.S. Environmental Protection Agency.  1975.  National Eutrophication
    Survey Methods 1973-1976.  Working Paper No. 175.  Environmental
    Monitoring and Support Laboratory. Las Vegas, Nevada, and
    Corvallis Environmental Research Laboratory, Corvallis, Oregon.
    91 pp.

Zand, S. M.  1976.  Indexes associated with information theory in water
    quality.  Journal WPCF.  48(8):2026-2031.
                                  10

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             APPENDIX.  SUMMARY OF PHYTOPLANKTON DATA
    This appendix was generated by computer.   Because it was only
possible to use upper case letters in the printout, all  scientific
names are printed in upper case and are not italicized.

     The alphabetic phytoplankton lists include taxa without species
names (e.g., EUNOTIA, EUNOTIA #1, EUNOTIA ?,  FLAGELLATE, FLAGELLATES,
MICROSYSTIS INCERTA ?, CHLOROPHYTAN COCCOID CELLED COLONY).   When
species determinations were not possible, symbols or descriptive
phrases were used to separate taxa for enumeration purposes.  Each
name on a list, however, represents a unique  species different from
any other name on the same list, unless otherwise noted, for counting
purposes.

    Numbers were used to separate unidentified species of the same
genus.  A generic name listed alone is also a unique species.   A
question mark (?) is placed immediately after the portion of a name
which was assigned with uncertainty.   Numbered, questioned,  or otherwise
designated taxa were established on a lake-by-lake basis; therefore
NAVICULA #2 from lake A cannot be compared to NAVICULA #2 from lake
B.  Pluralized categories (e.g., FLAGELLATES, CENTRIC DIATOMS, SPP=)
were used for counting purposes when  taxa could not be properly
differentiated on the counting chamber.
                                   11

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LAKE NAPE: DEEP CREEK LAKE
STORET NUMBER:  2402
                                              NYGAARD TPOPHIC STATE INDICES

                                        DATE   04 21 73  07 23 73  10 04 73
                                 MYXOPHYCEAN
                               CHLORCPHYCEAN
                                EUGLENCPHYTE
                                      DIATOM
                                    COMPOUND
0/01 0
1.00 E
1.00 E
0.20 7
3.00 E
05/0 =
04/0 E
0.11 7
1.67 E
15/0 E
2.00 £
1. CO E
0.33 E
0.50 E
6.00 E
                                            PALMER'S ORGANIC POLLUTION INDICES

                                        DATE   04 21 73  07 23 73  10 04 73
                                       GENUS
                                     SPECIES
02
03
01
00
01
00
                                         SPECIES DIVERSITY AND ABUNDANCE INDICES

                                        DATE   04 21 73  07 23 73  10 04 73
                      AVERAGE DIVERSITY
                         NUMBER OF' TAXfl
           NUMBER  OF SAMPLES CCMPOSITEC
                      MAXIMUM DIVERSITY MAXH
                        TOTAL DIVERSITY
         TOTAL  NUMBER OF INDIVIDUALS/ML
                      EVENESS COMPONENT
        MEAN NUMBER CF INDIVIDUALS/TAXA
       NUMBER/ML  OF MOST ABUNDANT TAXON
H
S
M
;H
c
N
J
L
K
2.56
14.00
4.00
3.81
2391.04
934.00
0*67
66.71
334.00
2.89
24.00
4.00
4.58
7742.31
2679.00
0.63
111.63
686.00
2.45
16.00
4.00
4.00
5902.05
2409.00
0.61
150.56
821.00

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LAKE NAME: DEEP CREEK LAK =
STOPE-T NUMBER: 2402
CONTINUED
TAXA

ANABAENA
ANKISTPCDPSMUS FALCATUS
APHANOCAPSA
ARTHROCESMtlS
ARTHRODESMUS INCUS
 V. RALFSII
ASTERICNELLA FORMOSA
 V. GRACILLIMA
CENTRIC CIATOM
CHRYSOCAPSA ? PLANCTCNICA
CRUCIGENIA TETRAPEDIA
CRYPTOMCNAS
DINCBRYON S5RTULARIA
CINCBRYQN SOCIALE
CINOFLAC-ELLATE
EUGLENA
FLAGELLATES
LYKGBYA
MELOSIRA #2
PELOSIRA #3
MELOSIRA DISTANS
HERISMOPEOIA
PICPOCYSTIS
MICRGCYSTIS INCERTA
NAVICULA HI
NAVICULA #2
PFDIASTRtM TETRAS
 V. TETRAODON
PEKNATE CIATCM #1
PERIDINIUM HISCONSINENSF.
SCENEDESPUS ABUNCANS
SCENEDESHUS BIJUGA
                                                  04 21 73
                                                                   07 23 73
                                                  10  04  73
FORM
FIL
CEL
ecu
CEL
CEL
CEL
CEL
CEL
COL
CEL
CEL
CEL
CEL
CEL
CEL
FIL
CEL
CEL
CEL
COL
COL
COL
CEL
CEL
COL
CEL
CEL
COL
COL
ALGAL
UNITS
S TC PER ML
1

51 7.4| 69



2



3
4



1
















X
13.7 128



13.71 128
17.91 167
1 X
5.21 49
1.11 10
35.81 334
















X





X





ALGAL
UNITS
S *C PER ML







5


1



2


3











2. 8| 74

1.2



0.4
25.6
0.4

22.1



19.7


4.3
0.8
7.3





0.4
2.8



32


X
10
6S6
10

591

X
X
528
X
X
116
21
195
X



X
10
74
X
X
ALGAL
UNITS
S JC PER ML





5




2



4


3











0.9




4.1



2.6
2E.7



13.1


9.6



0.9







21


X

98

X
X
63
692


X
315

X
231



21
X


X




-------
LAKE NAME: DEEP CREEK LAKE
SECRET NUMBER: 2402
74XA

STEPHANCCI?CUS DUBIUS
SYNE OR A
SYNUPA
SYNURA 1
TABELLARIA
TABELLARIA FENESTRATA
CONT!NUED
                                                  04 21 73
                                07 23 73
10 04 73


FORM
CEL
CEL
CEL
CEL
CEL
CEL


S






ALGAL
UNITS
*C PER ML
1
1 X

1
1 X
5.21 49


S *C
4(10.0
1
1
1
1
1 2.4
ALGAL
UNITS
PER ML
269

1
1 X

63
ALGAL
UNITS
S *C PER ML
... .

1 1
1 6.11 147
1 1
1 I
1(34.11 821
  TOTAL
                                                     934
                                  2679
                                                                                       2409

-------
LAKE NAME:  LIBERTY  RES.
SECRET NUMBER:  2403
                                              NYGAARD TROPHIC STATE INDICES

                                        DATE   04 11 73  07 20 73  10 01 73
                                 MYXQPHYCEAN
                               CHLOROPHYCEAN
                                EUGLENOPHYTE
                                      DIATOM
                                    COMPOUND
0/0 0
02/0 E
0/02 ?
0.30 ?
05/0 E
2.50 E
0.50 7
0,33 P
1. 00 E
6.50 E
05/0 E
0/0 0
0/05 ?
0.86 E
11/0 E
                                            PALMER'S ORGANIC POLLUTION INDICES

                                        DATE   04 11 73  07 20 73  10 01 73
                                       GENUS
                                     SPECIES
     01
     00
03
00
02
00
                                         SPECIES DIVERSITY AND ABUNDANCE INDICES
                                        DATE
                      AVERAGE DIVERSITY
                         NUMBER OF TAXA
           NUMBER OF SAHPLES COMPOSITED
                      MAXIMUM DIVERSITY MAXH
                        TOTAL DIVERSITY
         TOTAL NUMBER OF INDIVIDUALS/ML
                      EVENESS COMPONENT
        WEAN NUMBER CF INDIVIDUALS/TAXA
       NUMBER/ML OF MOST ABUNDANT TAXON
04 11 73  07 20 73  10 01 73
H
S
M
;H
D
N
J
L
K
2.08
20.00
4.00
4.32
3178. 24
1528.00
0. 48
76.40
765.00
2.19
2B.OO
4.00
4.81
7513.89
3431.00
0.45
122.54
1643.00
2.83
22.00
4.00
4.46
11090.77
3919.00
0.63
178.14
1421.00

-------
LAKE NAME: LIBERTY RES.
STCRET NUMBER: 2403
CONTINUED
                                                  04  11 73
                                                                    07  20  73
                                                                                     10  01  73
TAXA

ANABAENA
ANKISTRODESMUS
ASTERIONEULA FORMOSA
 V. GRACILLIMA
GERANIUM HIRUNOINELLA
COELASTRUM
COELOSPHA?RIUM NAEGELIANUM
CRYPTOMONAS
CRYPTOMCNAS #1
CRYPTOMONAS 02
CYANOPHYTAN FILAMENT
CYCLOTELLA
CYCLOTELLA STELLIGERA
CYMBELLA
DINOBBYON
DINOBRYCN BAVARICUM
DINCBRYON DIVERGENS
CINOFLAC-ELLATE
EUGLENA #1
EUGLENA #2
FLAGELLATE #1
FLAG ELLA-IPS
FRAG ILARIA
FRAGILARIA CROTCNENSIS
CCMPHOSPHAERIA ? LACUSTRIS
LYNGBYA
VALLlMCNAS PSEUCCCOFONATA ?
MELOSIRA #2
fELOSIRA DISTANS
MELOSIRA ITALICA
MEPISMOPEDIA
MICRCCYSTIS
FORM
FIL
CEL
CEL
CEL
COL
COL
CEL
CEL
CEL
FIL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
COL
FIL
CEL
CEL
CEL
CEL
CCL
COL
S


4




2
5











3







1


K

0.0
0.0




16.4
0.0



0.0







50.1







21.3


ALGAL
UNITS
PER ML

10
42




251
63


X
21
X






765







325


S






5















1





2


*C
1.4



1.4

4.2




1.4




1.4



1.4

47.9





23.6


ALGAL
UNITS
PER ML
48


X
48
X
143


X
X
A8


X

48
X
X
X
48

1643
X

X

X
610


S









I

4








5
2






3


*C
0.5





5.1


36.3

9.2




0.5



6.4
22.0


2.8

1.8

6.9
1.4

ALGAL
UNITS
PER ML
18





198


1421

360
X


X
18



249
863
X

108
1
72
1 X
1 270
54
1 X

-------
LAKE NAME: LIBERTY RES.
STOPET NUMBER: 2403
CCNTINUED
                                                  04  11  73
                                                                    07  20  73
                                                                                     10 01 73
TAXA

NAVICULA
NAVICULA CRYPTOCEPHALA
NAVICULA HAMBER6TI
NITZSCHIA
OSCILLATORIA
PENMATE DIATOMS
PERIDINIUM
SELENASTRUM ?
STAUPASTRUM #1
S^AURASTRUM #2
STEPHANOOISCUS
STEPHANCDISCUS ? OUBIUS
STEPHANODISCUS DUBIUS
SYNEDRA
SYNEORA ?
SYNEORA «1
SYNECRA #2
SYNEORA ULNA
TABELLARIA
TABELLARIA FENESTRATA
TABELLARIA FLOCCULCSA
FORM
CEL
CEL
CEL
CEL
FIL
CEL
CEL
COL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
S





















XC
1
0.0








0.0






0.0



ALGAL
UNITS
PEP ML

31
X
X


X
X


10



X
X

10

X
X
S











4




3




SIC











3.5




13.9




ALGAL
UNITS
PER ML
X
X


X

X

X
X

119

X


476




S





















*C





1.8






5.1



0.5




ALGAL
UNITS
PEP. ML

X



72





X
198



13

X


  TOTAL
                                                     1528
                                   3431
                                                                                       3919

-------
                  LAKE  NAHE:  LCCH  RAVEN  RES.
                  STORET  NUMBER; 2409
                                                                NYGAARD TRCPHIC STATE INDICES

                                                          DATE   04 11 73  07 21 73  10 Cl 73
                                                   MYXQPHYCEAN
                                                 CHLOROPHYCEAN
                                                  EUGLENOPHYTE
                                                        DIATOM
                                                      COMPOUND
02/0 E
0/0 0
0/02 ?
0.42 E
OT/0 E
6.00 E
6.00 E
0/12 ?
1. 00 5
16.0 E
7.00 E
4.00 E
0/11 ?
1.00 E
16.0 E
00
    PALMEP'S ORGANIC POLLUTION INDICES

DATE   04 11 73  07 21 73  10 01 73
                                                         GENUS
                                                       SPECIES
            07
            00
02
00
01
00
                                                           SPECIES  DIVERSITY AND ABUNDANCE INDICES
                                                          DATE

                                        AVERAGE'DIVERSITY
                                           NUMBER  OF  TAXA
                            NUMBER OF  SAMPLES COMPOSITED
                                        MAXIMUM  DIVERSITY  MAXH
                                         TOTAL  DIVERSITY
                          TOTAL  NUMBER OF  INDIVIDUALS/ML
                                        EVENESS  COMPONENT
                          MEAN  NUMBER CF  INDIVIOUALS/TAXA
                         NUMBER/ML OF MOST  ABUNDANT TAXON
       04 11 73  07 21 73  10 01 73
H
S
M
:H
D
N
J
L
K
2.50
23.00
3.00
4.52
5265.00
2106.00
0.55
91.57
712.00
1.76
25.00
3.00
4.64
9396.64
5339.00
0.33
213.56
3659.00
2.36
25.00
3.00
4.64
5354.84
2269.00
0.51
90.76
1193.00

-------
LAKE NAME:  LCCH PAVEN RES.
STORET NUMBER: 2408
CONTINUED
                                                  04 11  73
                                07 21 73
                                                                                    10 01 73
TAXA

ANABAENA #1
ANABAENA #2
ASTERICNELLA FORMOSA
ATTHEYA
CENTRIC DIATOM
CERATIUM HIRUNDINELLA
CHROOCCCCUS LINNETICUS
COELASTRUM RETICLLATUM
COELOSPHAERIUM
COELOSPHAERIUM NAEGELIANUM
CRUC1GENIA APICULATA
CRYFTOMONAS ?
CYCLOTELLA STELLIGERA
CYMBELLA
DINCBRYCN 6AVARICUM
DINCBRYCN OIVERGENS
DINOFLAGELLATE
FLAGELLATES
FRAGILARIA CROTCNENSIS
GCMPHOSPHAERIA LACUSTRIS
LYNGBYA BIRGEI
MELOSIRA *2
HELOSIPA #3
MELOSIRA DISTANS
MELOSIRA ITALICA
HICRCCYSTIS AERUGINOSA
NAVICULA
NAVICULA CAP1TATA
NAVICULA CRYPTQCEPHALA
NAVICULA VULPINA
NITZSCHIA PALEA
OOCYSTIS
FORM
FIL
FIL
CEL
CEL
CEL
CEL
COL
COL
COL
COL
COL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
COL
FIL
CEL
CEL
CEL
CEL
COL
CEL
CEL
CEL
CEL
CEL
CEL
S


2








4





3
I













*C


13.7

1.2






6.6




2.4
33.8
2S.3









0.6



ALGAL
UNITS
PER ML


394

25






140
X



51
712
597





X



12
X
X

S












2




4
1




3







5
S5C
1.1
1.1
0.6


0.6




0.6

10.7




3.9
63.5
0.6

1.7

7.3







1 3.4
ALGAL
UNITS
PER ML
60
60
30
X

30

X
X

30

570

X
X

210
3659
30

90

390

x
X




1 ISO
S









4







5
1



3

2







?C
1.5|
3.0





0.7

4.5


1.5




S.3
52.8



5.3

15.9







ALGAL
UNITS
PER ML
34
65



x
X
17

103


34
X

X

189
1198
X
X

120
X
360

X
X





-------
           LAKE NAME: LCCH  RAVEN  RES.
           SECRET NUMBER.: 2408
                                   CQNTINUED
ro
o
TAXA

CSCILLATCRIA
CSCULATORIA #1
PECIASTRUM SIMPLEX
 V. DUCDENARiUM
PEDIASTRUM TETRAS
 V. TETRAOCCN
PENNATE DIATOM
PENNATE CIATOM #1
PENNATE DIATOM #2
PENNAtE DIATOM #3
PERIDINIUM
SCENEOESPUS BIJUGA
SCENEDESMUS DISPAR
STAUPASTRUM
STAURONEIS ANCEPS
 F. LINEARIS
STEPHANCDISCUS
STEPHANQDISCUS ? DUBIUS
SVNEDRA
SYNEDRA ULNA
TABELLARIA
TETRAECRON MINIMUM
 V. SC'B'OBICt'LATUH

  TOTAL
                                                              04 11 73
                                                                               07 21 73
                                                                                                 10  01  73
FORM
FIL
FIL
COL
COL
CEL
CEL
CEL
CEL
CEL
COL
COL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL

S
5



















*C
3.0



0.6

0.6






1.8

1.2
1.2



ALGAL
UNITS
PER ML
63
X


12
X
12
X
X



X
38
X
25
25


2106
S




















*C

















>•


ALGAL
UNITS
PER ML

X

X





X

X





X
X
5339
S




















%C

C.7
2.6











3.0





ALGAL
UNITS
PER ML

17
59






X
X
X


69
X



2269

-------
                   LAKE NAKE:  JCHNSCN PCNO
                   STCRET NUMBER:  2409
                                                                 NYGAARC TROPHIC STATE 1NCICES

                                                           DATE   0* 10 73  07 20 73  09 28 73
                                                    MYXOPHYCEAN
                                                  CHLOROPHYCEAN
                                                   EUGLENOPHYTE
                                                         DIATOM
                                                       COMPOUND
2.00 E
1.00 E
0.33 E
0.27 ?
8.00 E
07/0 E
25/0 E
0.09 ?
0.20 ?
36/0 E
2.00 E
5.00 E
O.C7 7
1.50 E
9.00 E
ro
                                                               PALMER'S ORGANIC PCLLUTICN INDICES

                                                           DATE   04 10 73  07 20 73  09 28 73
                                                          GENUS
                                                        SPECIES
01
00
21
07
07
04
                                                            SPECIES DIVERSITY AND ABUNDANCE INDICES

                                                           DATE   04 10 73  07 20 73  09 28 73
                                         AVERAGE DIVERSITY
                                            NUMBER OF TAXA
                              NUMBER OF SAMPLES CCMPOSITED
                                         MAXIMUM DIVERSITY MAXH
                                           TOTAL DIVERSITY
                            TOTAL NUMBER OF INDIVIDUALS/ML
                                         EVENESS COMPONENT
                           MEAN NUMBER OF INDIVIDUALS/TAX A
                          NUMBF.R/ML OF HOST ABUNDANT TAXON
H
S
M
;H
D
N
J
L
K
3.28
32.00
2.00
5.00
2384.56
727.00
0.66
22.72
231.00
4.07
46.00
1.00
5.52
19552.28
4804.00
0.74
104.43
683.00
2.74
28.00
1.00
4.81
12475.22
4553.00
0.57
162.61
1422.00

-------
             LAKE NAME:  JCHNSON  PCNO
             STORET  NUMBER:  2409
                                   CONTINUED
ro
ro
                                                               04 10 73
                                                                                07 20 73
                                                                                                 09 28 73
TAXA

ACTINASTRUM HANTZSCHII
ANABAENA
ANABAENA #1
ANKISTPOCESMUS FALCATUS
ARTHRGDESML'S
CENTRIC DIATOM
CHLAMYDCMONAS
CLCSTERIUM
COCCONEIS PLACENTULA
COELASTPUM RETICULATUM
COELASTRUM SPHAERICUM
CQELOSPHAER1UH
COELOSPhAERIUM DUBIUM
CRUCI6ENIA APICULATA
CRUC1GENIA TETRAPEDIA
CPYPTCMCNAS
CRYPTOMONAS ?
CYANOPHYTAN FILAMENT
CYHBELLA
DACTYLOCOCCOPSIS
DINOBRYCN DIVF.RGENS
DINOFLAGELLATE
EUDORINA
EUDORINA ELEGANS
EUGLENA
EUGLENiA #1
EUNQTFA #1
EUNOTIA INCISA
FLAGELLATE #\
FLAGELLATE #2
FLAGELLATE #3
FLAGELLATES


FOPM
COL
FIL
FIL
CEL
CEL
CEL
CEL
CEL
CEL
CDL
COL
COL
COL
COL
COL
CEL
CEL
FIL
CEL
CEL
CEL
CEL
COL
COL
CEL
CEL
CEL
CEL
CEL
CEL
ALGAL
UNITS
S SEC PER ML















































2.6
2.6








X















19
19
X
X




2.61 19
1 X
5.21 38
5(15. SI 115
CEL 11131.81 231
CEL I
2.61 19
ALGAL
UNITS
S *C PER ML


X
41 6.91 330
1 I
51 7.81 377




























1
4.01 194



3.4
2.9
0.5


0.5

3.9







1.0
0.5





X

X
165
141
23
X
X
23

188






X
47
23





14.21 683
ALGAL
UNITS
S %C PER ML















2












1
5











2.4





25.8












31.2
6.2

10.1


X

X
X

X

107





1174

X




X

X



1422
284

462

-------
           LAKE NAME: JCHNSCN FCND
           STORET NUMBER: 2409
                                   CCNTINUED
PO
co
                                                             04 10 73
                                                                   07 20 73
                                                                                               09 28  73
TAXA

FRAGILARIA
FRAGILAPIA #1
FRAGILARIA #2
GOLENKIMA
GOLENKINIA RADIATA
GOPPHONEMA
GYPCSIGMA
KIRCHNERIELLA
LAGERHEIMtA
LAGERHEIMIA LONGISFTA
LUNATE CELLED COLONY
MELOSIRA #2
MELOSIRA #3
MELOSIRA DISTANS
MELOSIRA VARIANS
VERISMCPEDIA
flCRCCYSTIS AERUGINOSA
NAVICULA
NAVICULA RHYNCHCCEPHALA
 V. AMPHICEROS
NEIDIUM APICULATUM
 V. CONSTRICTUM
NITZSCHIA
CSCILLATORIA GEMINATA
PANDORINA HORUM
PEDIASTHUM BORYANUM
PEDIASTRUM DUPLEX
FEDIASTRUM TETRAS
 V. TETRAODON
PENNATE DIATOM #1
PHORMIDIUM MUCICOLA
PCLYEDRICPSIS SPINULCSA


FORM
cil
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CCL
CEL
CEL
CEL
CEL
COL
CCL
CEL
CEL
CEL
CEL
FIL
COL
COL
COL
COL
CEL
COL
CEL


S
"
2












4
















*C
1
10.61



2.6




2.6
5.2


5.2











2.6


ALGAL
UMTS
PER ML

77
X


19
X



19
39
X
X
38



X
X
X

X



19




S
















1




2









*C
6.4


0.5
2.9



0.5







10.8



6.4
8.8
0.5

0.5


9.3

ALGAL
UNITS
PER ML
306


23
141


X
23
X





X
518



306
1 424
23
1 X
1 23
I
1
1 x

1 448
1 X


S











3




4














XC











12.5




3.9










3.9

ALGAL
UNITS
PER ML
X



X






569


X

178
X




X




1 178
1

-------
z
o
o
            LAKE NAME: JCHNSCN PCND
            STOR'ET NUMBER: 2409
                                   CONTINUED
ro
                                                              04 10 73
                                                                               07  20  73
                                                                                                 09  28 73
TAXA

SCENEOESMUS
SCEN'EDESMUS #1
SCENEDESMUS #2
SCENEDESNUS #3
SCENEDESMUS ABUNDANS
SCENEDESMUS BIJUGA
SCENEDESMUS DISPAR
SCENEDESMIS GUTVilNSKII
SCENEDESKUS PROTUBERANS
SCENEDESMIS QUADRICAUDA
SCENEDESMUS QUADRICAUOA
 V. LCNGISPINA
SCHROEDERIA SETIGERA
STAUPASTRUM PUNCTULATUM
SURIRELLA
SYNEDRA #1
SYNEDPA DELICATISSIMA
SYKEORA RUPPENS
 V. SCOTIA
TA6ELLARIA #1
TETRAffcRON
TETPAECRON #1
TETRASTRUM HETERACANTHUM
TRACHELCMONAS
TREUBARIA
FORM
COL
COL
COL
COL
COL
COL
COL
CCL
COL
CCL
COL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
CEL
COL
CEL
CEL
S














3








*C





2.6








5.2








ALGAL
UNITS
PER ML





19






X
X
38

X
X



X

S























?C


1.0
1.0




3.5
3.4

1.0









0.5
0.5
ALGAL
UNITS
PER ML

X
47
47


X
X
23
165

47



X

X
X


23
23
S























%C




0.8




1.6









0.8
0.8


ALGAL
UNITS
PFR ML
X



36
X



71
X
X







36
36


              TOTAL
                                                                  727
                                                                                  4604
                                                                                                   4553

-------
                                   TECHNICAL REPORT DATA
                            (Please read Instructions on the reverse before completing)
1. REPORT NO.
 EPA-600/3-77-124
                                                            3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE

DISTRIBUTION  OF PHYTOPLANKTON  IN  MARYLAND LAKES
              5. REPORT DATE
               October  1977
                                                            6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
V.W. Lambou, F.A. Morris, R.W. Thomas, M.K. Morris, L.R.
Williams. W.D. Taylor. F.A. Hiatt, S.C. Hern. J.W.  Hilgert
t  DC a criDKyil Mfz nar? AMi"7 ATII"IM MAKAC AMr-» Annaccc
                                                            8. PERFORMING ORGANIZATION REPORT NO
». PERFORMING ORGANIZATION NAME AND ADDRESS
invironmental  Monitoring and Support Laboratory
)ffice of  Research and Development
J.S. Environmental Protection Agency
Las Vegas,  Nevada 89114
                                                            10. PROGRAM ELEMENT NO.
               1BA608
              11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
J.S. Environmental Protection Agency-Las Vegas, NV
Dffice of Research and  Development
Invironmental  Monitoring and Support Laboratory
Las Vegas, Nevada 89114
              13. TYPE OF REPORT AND PERIOD COVERED

               03-07-73  to ll-U-73	
              14. SPONSORING AGENCY CODE
                EPA/600/07
15. SUPPLEMENTARY NOTES
previously released in limited distribution as No. 684  in  the Working Paper  Series
For the National  Eutrophication Survey.
16. ABSTRACT
     This is a data  report presenting  the species and abundance of phytoplankton
in the 4 lakes sampled by the National  Eutrophication Survey in the State of
Maryland.  Results from the calculation of several water  quality indices are also
included (Nygaard's  Trophic State  Index, Palmer's Organic Pollution Index, and
 pecies diversity and abundance indices).
 7.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b.lDENTIFIERS/OPEN ENDED TERMS
                           c.  COSATI Field/Group
*aquatic microbiology
 lakes
*phytoplankton
 water quality
Maryland
lake eutrophication
Nygaard's trophic indices
Palmer's organic pollu-
 tion indices
Species diversity and
abundance indices
 06  C,M
 08  H
 13  B
 8. DISTRIBUTION STATEMENT

    RELEASE TO  PUBLIC
19. SECURITY CLASS (ThisReport)
    UNCLASSIFIED
21. NO. OF PAGES
     32
                                               20. SE
                                                            S/This page)
                                                                          22. PRICE
EPA Form 2220-1 (Rev. 4-77)   PREVIOUS EDITION is OBSOLETE

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