Ecological Research Series
EVALUATION OF  THE  ALGAL  ASSAY PROCEDURE
                                      Environmental Research Laboratory
                                      Office of Research and Development
                                     U.S. Environmental Protection Agency
                                             Corvallis, Oregon 97330

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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 five  series. These five broad
 categories  were established to facilitate further development and application of
 environmental technology.  Elimination of traditional grouping was consciously
 planned to  foster technology transfer and  a maximum interface in related fields.
 The five series are:

     1.    Environmental Health Effects Research
     2.    Environmental Protection Technology
     3.    Ecological Research
     4.    Environmental Monitoring
     5.    Socioeconomic Environmental Studies

 This report has been assigned to the ECOLOGICAL RESEARCH series. This series
 describes  research on the effects  of pollution on humans, plant  and animal
 species, and materials.  Problems are assessed for their long-  and short-term
 influences.  Investigations include formation, transport, and pathway studies to
 determine 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-600/3-76-064
                                            June 1976
  EVALUATION OF THE ALGAL ASSAY PROCEDURE
                     by

              Charles M. Weiss

University of North Carolina at Chapel Hill
           Chapel Hill, NC 27514
           Grant No. R800399-06-0
              Project Officer

             William E. Miller
Assessment and Criteria Development Division
Corvallis Environmental Research Laboratory
          Corvallis, Oregon 97330
    U.S. ENVIRONMENTAL PROTECTION AGENCY
     OFFICE OF RESEARCH AND DEVELOPMENT
CORVALLIS ENVIRONMENTAL RESEARCH LABORATORY
           CORVALLIS, OREGON 97330

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                             DISCLAIMER

This report has been reviewed by the Corvallis Environmental Research
Laboratory, U.S. Environmental Protection Agency, and approved for
publication.  Approval does not signify that the contents necessarily
reflect the views and policies of the U.S. Environmental Protection
Agency, nor does mention of trade names or commercial products consti-
tute endorsement or recommendation for use.

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                                   ABSTRACT
     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.

     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
productivity.

     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 measure
of total growth potential and the latter ambient growth potential.
                                     tii

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                               ACKNOWLEDGEMENTS
     This extended investigation carried out over a period of several years
involved the efforts of many staff and students of the Department of Environ-
mental Sciences and Engineering.  The samples collected for the field evaluation
were the particular responsibility of Terry Anderson, Mark Mason and Chris
Knud-Hansen.  Algal assays were carried out by Sandy Hoffman, Pat Appeldorn and
Chris Knud-Hansen.  Dr. Peter H. Campbell did the phytoplankton counts.  This
included a population analysis of the phytoplankton which will be reported
elsewhere.
                                     IV

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                            CONTENTS
Abstract                                                    iii
Acknowledgements                                             iv
Figure                                                       vi
List of Tables                                              vii
I     Introduction                                            1
II    Summary                                                 2
III   Conclusions                                             3
IV    Recommendations                                         4
V     Experimental Plan                                       5
VI    Methods and Procedures                                  9
VII   Results and Discussion                                 12
VIII  References                                             50
IX    Appendix                                               52

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                              FIGURE
Number
        Location and Nature of Surface Waters Sampled  in
           North Carolina and Adjacent Areas 	    8
                              VI

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                                 LIST OF TABLES
Number
       Surface Waters of North Carolina and Adjacent Areas
         Evaluated by Algal Assay 	
       Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays,  Mean Values Within Value
         Range of Indicated Parameter:   Inorganic Nitrogen
         (NH3+N02+N03) mg/m3
  3    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter:  Total Nitrogen - mg/m3  .  .    15

  4    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter:  Total Soluble-P - mg/m3 .  .    16

  5    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter:  Total-P - mg/m3 ......    17

  6    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter:  Ratio TN/TP ........    18

  7    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter:  Cell Density No. /ml ....    19

  8    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter:  Chlorophyll a, Turner
         Units  ..........................    21

  9    Physical, Chemical and Biological Parameters,  All Lake
         Samples Used in Algal Assays, Mean Values Within Value
         Range of Indicated Parameter :  Productivity - mgC/m3/hr  . .   22

 10    Growth of Selenastrum capricornutum in NAAM (14 Days, 24°C,
         400 f.c.); Mean Values of Biomass and Unit Absorbance  .  .    23

 11    Growth of Selenastrum capricornutum in Lake Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus ......................    25


                                      vii

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                            LIST OF TABLES Continued
Number                                                               Page

  12   Growth Response of Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus	    26

  13   Growth Response of Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus	    27

  14   Growth Response of Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus	    28

  15   Growth Response of Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus	    29

  16   Growth Response of Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus	    30

  17   Growth Response of Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes of Nitrogen
         and Phosphorus	    31

  18    Growth Response of  Selenastrum capricornutum in Lake  Samples,
         Controls and on Addition of Nutrient Spikes  of Nitrogen
         and  Phosphorus	    32

  19    Growth Response of  Selenastrum capricornutum in Lake  Samples,
         Controls  and on Addition of Nutrient  Spikes  of  Nitrogen
         and  Phosphorus	    33

  20    Growth  Response  of  Selenastrum capricornutum in Lake  Samples,
         Controls and  on Addition  of Nutrient  Spikes  of  Nitrogen
        and Phosphorus	    34

 21   Growth Response of Selenastrum  capricornutum in Lake  Samples,
        Controls and on Addition  of Nutrient  Spikes  of Nitrogen
        and Phosphorus	    35

 22   Correlations of Seeded Control Growth and Nutrient
        Concentrations Following Pre-Treatments  	    37

 23   Correlations of Seeded Control Growth and Nutrient
        Concentrations Following Pre-Treatments  	    38

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                            LIST OF TABLES Continued
Number
  24   Correlations of Seeded Control Growth and Nutrient
         Concentrations Following Pre-Treatments 	    39

  25   Correlations of Seeded Control Growth and Nutrient
         Concentrations Following Pre-Treatments 	    40

  26   Correlation of Growth in Non-Spiked Samples of Phosphorus
         Limited Waters,  Autoclaved Pretreatment, with Sample
         Nutrients and Comparison to Original Lake Nutrient
         Concentrations  	    41

  27   Correlation of Growth in Non-Spiked Samples of Phosphorus
         Limited Waters,  Filtered Pretreatment,  with Sample
         Nutrients and Comparison to Original Lake Nutrient
         Correlations  	    42

  28   Correlation of Growth in Non-Spiked Samples of Nitrogen
         Limited Waters,  Autoclaved Pretreatment, with Sample
         Nutrients and Comparison to Original Lake Nutrient
         Concentrations	    43

  29   Correlation of Growth in Non-Spiked Samples of Nitrogen
         Limited Waters Filtered Pretreatment, with Sample
         Nutrients and Comparison to Original Lake Nutrient
         Concentrations  	    44

  30   Correlation of Growth in Non-Spiked Samples of Phosphorus
         & Nitrogen Limited Waters, Autoclaved Pretreatment, with
         Sample Nutrients and Comparison to Original Lake Nutrient
         Concentrations  	    45

  31   Correlation of Growth in Non-Spiked Samples of Phosphorus
         & Nitrogen Limited Waters, Filtered Treatment, With
         Sample Nutrients and Comparison to Original Lake Nutrient
         Concentrations  	    46

  32   Relationship of Control Growth, Seeded Senenastrum
         capricornutum in Autoclaved and Filtered Waters and
         Nutrient Levels of Original Raw Water  	    47

  33   Nutrient Limitation of Algal Assayed Samples, Control
         Biomass and Original Lake Nutrient Quality  	    49
                                       IX

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                                    SECTION I

                                  INTRODUCTION
     The problem of man induced eutrophication in the United States has led to
 the development of bioassay procedures for establishing both quantitative and
 qualitative response to the major algal nutrients.  It has been particularly
 important that these determinations be based upon a reference procedure so that
 effective regional comparisons could be carried out.  With this in mind a joint
 industry government task force on eutrophication encouraged development and
 publication of provisional assay procedures for use in predicting the impact of
 nutrient elements and nutrient-synergistic compounds upon aquatic productivity in
 natural waters.1

     An intensive coordinated program of development and evaluation followed
 involving university and industrial laboratories.  Both batch or bottle test and
 continuous culture algal bioassays were evaluated as to their applicability for
 establishing limiting nutrient levels in surface waters.2'3  Following detailed
 definition of the growth constraints for the batch test, eight laboratories
 participated in a joint evaluation of the assay bottle procedure as reported by
 Weiss and Helms.   The same water samples were distributed to each of the eight
 laboratories and batch tests were carried out using a standardized assay protocol.
 This interlaboratory comparison established that whereas there was a low level of
 precision between the results of the participating laboratories, variation being
 over 30%, within any one laboratory the precision was considerably better.
 Unknown systematic errors may have contributed to the magnitude of difference in
 biomass grown at the several laboratories.

     It was stated in the original industry/government task force recommendation
 on the algal assay procedure that when the developmental phase was completed an
 extensive field evaluation should be undertaken.  Without this correlation to the
 conditions in nature, the true relationship of the results obtained from the
 assay procedure would still remain uncertain.  One of the immediate and very
 useful applications of the algal assay has been to establish the extent of
 response of batch cultures to environmental safety evaluations of raw materials
 that might be considered for use in consumer products, Sturm and Payne,5 Payne,5
 Mitchell and Buzzel.7  Examples of algal response in natural systems using the
 batch assay to establish trophic levels and identification of growth limiting
 nutrients have been used by Toerien and Steyn,^ Francisco and Weiss,^ Steyn et
_al.10 Miller et^ al. ,n Payne,12 Doemel and Brooks,13 and Greene e_t al.ltf

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                                     SECTION  II

                                      SUMMARY
      The  345  algal  assays  of  surface water samples from lakes, rivers and
 impoundments  of  North  Carolina  carried out over a period of about five years
 included  the  pretreatment  procedure of autoclaving or membrane filtration.  This
 step  destroyed or removed  living  algal cells and permitted reseeding with a
 specific  test species,  Selenastrum capricornutum.  Subsequent growth under
 controlled  temperature and  light  conditions permitted comparison of biomass grown
 in  the pretreated (autoclaved or  filtered) media and that in waters to which
 known quantities of phosphorus  and nitrogen were added.  The growth in the
 spiked samples provided an indication of whether phosphorus, nitrogen or both
 might be  growth  limiting in the original water.  The growth in the unspiked water
 also  provided reference to  the  scale of ambient nutrient levels and the trophic
 state of  the  original  surface water source.

      The  control algal  biomass  grown in water with indicated phosphorus
 limitation, 104  assays  after  autoclaving as pretreatment, had a mean concentra-
 tion  of 5.31  mg/1 whereas filtered phosphorus limited waters, 191 assays, had a
 mean  biomass  of  1.37 mg/1.  Nitrogen limited water after autoclaving, 132 assays,
 had a mean biomass  of  12.15 mg/1  and 9.81 mg/1 following filtration as the
 pretreatment.  Where both phosphorus and nitrogen were indicated as limiting
 following autoclaving,  105 assays, the mean biomass was 4.70 mg/1 and 0.92 mg/1
 after filtration.

      The implication of these findings is that the autoclaving pretreatment
 procedure probably provides a better indication of potential growth capability
whereas the filtered samples  reflect the growth capability of ambient nutrient
 levels.

     A consistent pattern of  the ratio of the soluble nitrogen and phosphorus
components to the indicated limitation of either one, the other, or both of these
algal nutrients suggest that this ratio can be used to define which nutrient is
limiting for the particular body of water.

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                                   SECTION III

                                   CONCLUSIONS
     The growth of the reseeded test alga, Selenastrum capricornutum, in water
samples following pretreattnent either by autoclaving or filtration provides a
good indication of the total growth potential by the first procedure and ambient
growth potential by the second.  The growth response of samples to which spikes
of phosphorus and nitrogen have been added established the nature of the growth
limiting nutrient.  Reference to original nutrient levels clearly indicated
ranges of nitrogen to phosphorus ratios necessary for optimum support of the test
organism.  The determination of the N/P ratio appears to provide the same
information as derived from the more complicated assay spiking procedure
providing other essential nutrients or inhibitors are not growth limiting.

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                                   SECTION IV

                                 RECOMMENDATIONS
     The standard assay procedure could be modified to limit the determination
to biomass grown in reseeded samples following autoclaving and filtration.   The
determination of whether the waters are limited in their growth potential by the
relative quantity of phosphorus or nitrogen can be satisfactorily determined from
the ratio of the total soluble nitrogen and phosphorus components if other
constituents are not growth limiting.

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                                    SECTION V

                                EXPERIMENTAL PLAN
     One of the basic problems inherent in the batch algal assay procedure is
the necessary pretreatment required of the water sample, either filtration
through a 0.45y membrane filter or autoclaving followed by filtration to remove
particulate and insoluble materials.  Either pretreatment is essential in order
to remove or destroy living algal cells so that reseeding with the test alga is
possible.  Using filtration, the assumption is made that all remaining nutrient
components in solution would reflect the growth limiting nutrient:  regulating
further natural biomass production.  In the use of autoclaving, which also
solubilizes nutrient materials, algal growth would reflect the total nutrient
potential assuming temperature, light and other growth factors were not limiting.
In either case if the assay is meaningful the magnitude of growth of the reseeded
sample should relate to the trophic state or the limiting nutrient potential of
the original water sample.  In this field evaluation algal assays were used to
establish the relationship between the existing trophic or nutrient level of 44
different bodies of water, sampled at several locations, as well as several
frequencies, for a total of 345 assays.  The location and type of the sampled
waters is shown in Figure 1.  The name, identification code and size with
approximate magnitude of pollution entering each body of water is presented in
Table 1.

     TABLE 1.  SURFACE WATERS OF NORTH CAROLINA AND ADJACENT AREAS EVALUATED
               BY ALGAL ASSAY

                                                              Pollution Sources
Type and Name
Surface                     Direct
 Area     Mean    Stations   Point   Inflowing
(Acres) Depth-ft. Sampled  Discharge  River**
  Natural Lakes

 1 Black
 2 Jones
 3 Mattamuskeet
 4 Phelps
 5 Salters
 6 Singletary
 7 Waccamaw
 8 White

  Impounded Cooling Lakes

 9 Belews
10 Hyco
 1,418
   224
30,000
16,000
   315
   572
 8,938
 1,068
 3,700
 3,750
 6*
 6*
 4*
 6*
 6*
 6*
 6*
 8*
50
21
2
2
1
1
2
2
2
2
2
2
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
                                                                  (Continued)

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TABLE 1.  SURFACE WATERS OF NORTH CAROLINA AND ADJACENT AREAS EVALUATED
          BY ALGAL ASSAY Continued
Pollution Sources
Type and Name
Water Supply Impoundments
11 University
12 Michie
Hydroelectric Impoundments
Catawba River
13 James
14 Rhodhiss
15 Hickory
16 Lookout Shoals
17 Norman
18 Mt. Island
19 Wylie
20 Fishing Creek
21 Wateree
Yadkin River
22 W. Kerr Scott (Flood Control)
23 High Rock
24 Tuckertown
25 Badin
26 Tillery
27 Blewett Falls
Roanoke River
28 John H. Kerr
29 Gas ton
30 Roanoke Rapids
Surface
Area Mean
(Acres) Depth-ft.

200
507


5,510
3,515
4,110
1,270
32,510
3,235
12,455
3,370
13,710

3,980
15,180
2,529
5,973
5,000
2,500

48,900
22,000
4,900

9
25


46
21
31
24
34
18
22
17
23

38
16
17
24
34
36

34
18
16
Stations
Sampled

1
1


4
6
3
3
6
3
8
2
5

3
3
3
3
7
3

11
3
3
Direct
Point
Discharge

No
No


Yes
Yes
Yes
No
No
No
Yes
No
No

No
No
No
Yes
No
Yes

Yes
No
No
Inflowing
River**

No
No


No
No
No
No
No
No
No
Yes
Yes

No
Yes
No
No
Yes
No

Yes
No
No
                                                            (Continued)

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     TABLE 1.  SURFACE WATERS OF NORTH CAROLINA AND ADJACENT AREAS EVALUATED
               BY ALGAL ASSAY Continued
                                                              Pollution Sources
                                  Surface                     Direct
                                   Area     Mean    Stations   Point   Inflowing
                                  (Acres) Depth-ft. Sampled  Discharge  River"*
Type and Name
 Old Millponds

31 Crystal (1885)***      '          100      6*
32 Davies (1850)                     50      6*
33 Finches (1875)                    20      6*
34 Hodgins (1871)                   100      5*
35 Jackson (1885)                    75     10*
36 Johns (1840)                     125      8*
37 Jones (1810)                      75      3*
38 Lytches (1870)                   325      7*
39 McKensie (1860)                   50      7*
40 McNeils (1870)                   100      4*
41 Monroe (1825)                     70      5*
42 Orton (1810)                     500      8*
43 Tull (1875)                      180      6*

 River Segments

44 Chowan
   (U.S. 13 to Albemarle Sound)
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
                                                       1
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
                                                                Yes
   * Estimated.
  ** Yes  if  river  carries  pollution discharges from upstream communities  with
      no intervening  impoundments.
 *** Year of construction.

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OO
                                                 Natural Lakes




                                                 Millponds




                                             (_)  Water Supply Impoundments




                                             /\  Hydroelectric Impoundments



                                             V7  Cooling Water Ponds





                                                 River Segment



                                             SEE TABLEfDFOB LAKE IDENTIFICATION CODE
             Figure 1:    Location and Nature of Waters Sampled  in North Carolina  and  Adjacent Areas  for  Algal  Assays.

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                                   SECTION VI

                             METHODS AND PROCEDURES
     In order to accommodate the considerable number of samples being processed
over the several years of this study and to stay within the recommended
procedures of the "bottle test" the growth response of the reseeded samples were
determined only at the end of a fixed period of incubation.  It has been shown in
the developmental steps of the assay procedure that biomass achieves a level of
growth proportionate to available nutrients in 10 to 14 days with occasional
samples, depending on the particular character of the water, requiring upwards of
21 days to achieve the growth plateau.2  For this field evaluation the assay
protocol was standardized as follows:

     1.  Each raw water sample was split into two portions - one for auto-
claving and one for filtration (0.45y filter) as pretreatment procedures.

     2.  Each of the samples following pretreatment was divided into subsets
of three portions, each 60 ml in 250 ml Erlenmeyer flasks, and spiked to provide
a final additional concentration of 5 and 50 yg/1 of phosphorus, 75 and 750 yg/1
of nitrogen, 5 yg/lP plus 75 yg/lN, and 50 yg/lP plus 750 ug/lN.  In addition
another set of three flasks was used as a nonspiked control.

     3.  Each flask was seeded with 1,000 cells/ml of Selenastrum capricornutum
previously washed and resuspended in distilled water to adjust to the inoculation
concentration.  The inoculum was cell culture grown  10-21 days in New Algal
Assay Media (NAAM).2

     4.  The seeded flasks were placed in an incubator at 24°C+2.0°C under
400 ft. candles of "daylight" fluorescent illumination.

     5.  Incubation was for 14 days with a daily swirling of the cutlure flasks.

     6.  The standard control media NAAM, in a replicate set of flasks, and
seeded from the same cell suspension of _S_. capricornutum was incubated for the
same period as each assay set.

     7.  At the end of the growth period the optical density of the cell
suspension was determined using a Bausch. and Lomb Spectronic 20 colorimeter at a
wavelength of 640 my.  In addition the total biomass grown in the NAAM control
samples was optically measured, filtered and weighed and used to determine the
unit absorbance for the particular growth conditions of that incubation set.

     8.  The unit of absorbance of the NAAM control for each incubation set was
used to convert the unit absorbance of the spiked samples and controls to
biomass.

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      The requirement of maintaining extensive records over a relatively long
 period of time on the algal assays led to systematizing the procedures for
 computer conversion of data.  Data record forms were designed to accommodate the
 basic information of each water sample from which data cards were punched,
 programs written for data analysis and a good deal of the computational work
 processed by computer.

      At the time of collection of the original water sample on which the algal
 assay was to be carried out, a broad spectrum of limnological parameters were
 also determined.  From these the original trophic state could be established for
 comparison to the nutrient characteristics as determined by the algal assay.

      All analyses of the several nitrogen and phosphorus species were made using
 Technicon Autoanalyzers.  For the purposes of this investigation the nitrogen and
 phosphorus forms are defined or were determined as follows:

      NHo-N            Ammonia nitrogen.

      NC^+NOg-N        One analysis which includes oxidation of N0£ to N(>3 and the
                       total determined.

      Inorganic-N      The sum of NH3-N and NC^+NC^-N.

      Kjel-N           Kjeldahl nitrogen.

      Total-N          N02+N03-N plus Kjel-N.

      PO^-P             Orthophosphate phosphorus,  soluble reactive phosphorus.

      T-Sol  P          Total soluble  phosphorus.   The  total fraction that passes
                       the 0.45 micron filter  and  is digested  by autoclaving  with
                       persulfate.

      T-Particulate P   Total-P minus  total  soluble P.

      T-P               Total Phosphorus.  The  total raw  sample digested  by
                       autoclaving  with persulfate.

      Phytoplankton cell  counts  were  determined on live  samples,  concentrated by
centrifugation with a  clinical  centrifuge.  The resuspended concentrate was
transferred  to a microscope slide  and sealed  under a 22x22mm  cover  glass.
Transects were counted at  500X  and specific identifications made  at  1250X.   Cell
density was  calculated from the cover glass area,  area  of  transects  counted  and
original volume concentrated.

      The chlorophyll a. determinations were made on a Turner photo-fluorometer
equipped with a photo detector  that  enhanced  its  sensitivity  to chlorophyll
fluoresence.  Comparison of  the values determined  with  a Turner photo-fluorometer
and the standard acetone extraction procedure showed a  high level of  correlation.
However, since the conversion factor appeared to vary somewhat from  lake  to  lake
the Turner scale unit has been used in this report as the  chlorophyll unit.  An
approximate conversion to chlorophyll a_ mg/1  can be made by multiplying Turner
units by 0.44.


                                        10

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     Since it was realized that determination of in situ primary productivity
would not be logistically feasible in all cases, a procedure that would allow
for direct comparison of productivity between lakes was used.   Following suitable
experiments to define optimum conditions all raw water samples collected for
productivity determinations were returned to the laboratory in polyethylene
bottles and stored overnight in the dark at room temperatures.  The next day a
light/dark bottle set (2 light, one dark) were filled with the raw water sample
and after initial dissolved oxygen determinations incubated at 24°C and 400 f.c.
for six hours.  The rate of carbon fixed per hour was computed from the net
oxygen production.  The same samples returned to the laboratory for primary
productivity determinations also provided the water source for chlorophyll a_
measurements and plankton counts.
                                       11

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                                    SECTION VII

                               RESULTS AND DISCUSSION


 NUTRIENT CHARACTERIZATION OF NORTH CAROLINA LAKES

 Limnological Parameters

      Sampling for algal assay has coincided in most instances  with other water
 quality investigations of North Carolina waters.   Results  of some assays have
 already been used to define algal growth conditions of specific  locales or  to
 help define trophic states.15»16>17>18  At time of sampling a  variety  of
 limnological parameters were also defined.  In general samples for algal assay
 were taken either from mid-epilimnetic or Secchi  disc  depth, the choice
 approximating the mid-depth of the euphotic zone.   The mean values of  those
 parameters that either define the conditions for  algal growth  or the result of
 algal growth have been assembled in Appendix A for each lake or  where  meaning-
 ful for stations or sets of stations within a lake.

      The data has also been classified into two seasonal periods,  April through
 November and December through March.   The experience of sampling North Carolina
 waters has shown that these two seasonal segments  generally define, a  more  active
 growing season,  in the case of the former and a winter season  of somewhat lesser
 growth activity.   However,  the difference between  the  two  seasons, with respect
 to algal growth,  is often quite small.   Light is probably  the  overall  principal
 limiting factor  rather than temperature.

      Belews  Lake  and Lake Hyco,  cooling pond impoundments  are  each represented
 by two stations  at points on the lakes,  which describe different environmental
 conditions.   University  Lake and Lake Michie,  water  supply impoundments were
 sampled  only at  one station near the  water supply  intake.   Of  the  Catawba River
 impoundments, Lakes James,  Rhodhiss,  Norman,  Mt. Island, Fishing Creek and
Wateree  are  limnologically  described  by  averaging  data of  more than one station
 on each  lake.  Two adjacent impoundments,  on the Catawba River,  Hickory and
 Lookout  Shoals had their data combined  into one profile whereas  the data for Lake
Wylie  is presented by separate station  sets representing both  organic  and thermal
pollution  conditions.  The  Yadkin River  impoundments,  Kerr Scott,  High Rock,
Tuckerton, Badin,  Tillery and  Blewett  Falls are each described independently
whereas  the  large  flood  control  and hydro-power impoundment, John  H. Kerr was
sampled  and  analyzed  by  sets  of  stations  representing  changes  in water quality
along its  two flow-through  gradients.  Similarly the data  from the Chowan River
was analyzed by sets  of  stations  representing points of change in water quality.
All the small mill ponds  and  the  few natural  lakes, generally represented by one
sample, are  summarized independently.  Generally lakes of  similar nature or
location are arranged on  the same page in  order to facilitate comparison.
                                       12

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Nutrient Profiles

     The range of values for nutrient levels and biological parameters within
the two seasonal segments of the year, and the associated physical and chemical
determinations have been described for the assayed waters.  This was accomplished
using sorting procedure on the card punched limnological data for each lake.   A
first step would be to arrange the nearly 350 data cards in rank order for one
of the nutrient components.  For example, for inorganic nitrogen, the lowest
value of the entire series was 20 mg/m3 and the highest 1245 mg/m3.  The entire
set of inorganic nitrogen values was then divided into four subsets having ranges
of 20-120 mg/m3, 130-245 mg/m3, 250-400 mg/m3 and the highest 500-1245 mg/m3.  A
suitable computer program then selected out all lake samples in each of the five
ranges, as defined by the quantity inorganic nitrogen, and calculated mean
values for all other variables within that set, Table 2.  In the April-November
growing season, the 68 samples which occurred in the range of 20-120 mg/m3
inorganic nitrogen had an average water temperature of 25.2°C, an average Secchi
depth of 1.5 m, an ammonia nitrogen concentration of 35.5 mg/m3, etc.  If for
some reason the data set was incomplete for a particular parameter the actual
number of samples used in the average is identified in a parenthesis following
the value.  For example the mean value of 37.1 Turner units for chlorophyll a_
was defined by averaging 53 samples rather than the total number of 68 for that
seasonal set.  This method of data analysis established that over the five ranges
of inorganic nitrogen the total soluble phosphorus increased from 11.8 mg/m3, the
mean value for those water samples which had inorganic nitrogen concentrations in
the range of 20-120 mg/m3 to a total soluble phosphorus concentration of
54.4 mg/m3 associated with the highest range of inorganic nitrogen.

     Of most significance to the ultimate concern of this study, the relationship
of the algal assay to the trophic state of the raw waters, would be the three
biological determinations.  Phytoplankton cell density, chlorophyll a., and
productivity and their relationship to the ranges of the nutrient levels.  As an
example the cell density, Table 2, decreased systematically with increase in
inorganic nitrogen concentration.  By contrast chlorophyll ji remained relatively
constant throughout the range of concentrations but productivity appeared to have
a peak value in midrange.  All these changes were in samples taken in the April-
November growing season.  Such comparisons can be made with other parameters and
other related components and provides an opportunity to establish the relation-
ships of the mean values of various associated parameters.

     The sorting and averaging procedure was carried out for total nitrogen,
Table 3; total soluble phosphorus, Table 4; total phosphorus, Table 5; and the
ratio of total nitrogen to total phosphorus, Table 6.  It should be noted that in
each instance the particular values used to define the upper and lower limit of a
range was an actual determination and not an arbitrary division of the total rank
of values.  Thus there might be a gap between upper and lower limits of ranges
since the omitted values did not actually occur in the total range of values.

     A similar sorting procedure was carried out for the three biological
determinations, phytoplankton, cell density, chlorophyll a^, and productivity to
determine the mean values of nutrients that were associated with the ranges
selected for averaging.  The data of Table 7 examines the total range of cell
density from a low of 62 cells per ml to the maximum value of 82,671 cells/ml,
in five sets.  In the April-November growing season the soluble components of

                                        13

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                    TABLE 2.   PHYSICAL,  CHEMICAL AND BIOLOGICAL PARAMETERS, ALL LAKE
                              SAMPLES USED IN ALGAL ASSAYS,  MEAN VALUES WITHIN VALUE
                              RANGE OF INDICATED PARAMETER:   INORGANIC NITROGEN (NH^+NO?NO-^)  mg/m3

Sample Period
Range of Values mg/m^
N
Water Temp. °C
Secchi Depth-m
NH3-N*
Inorganic-N
Kjel-N
Total-N
PO.-P
4
T-Soluble P
T-Particulate P
Total-P
TN/TP
Cell Density no. /ml
Chlor a Turner Units
Productivity mgC/m3/hr.
April - November
20-120
68
25.2
1.5
35
30
64
341
361
7.3
11.8
15.0
26.5
17.6
6444
37.1(53)
36.3(26)
130-245
65
19.1
1.2
59
119
182
318
405
11.5
18.7
15.8
35.5
18.5
3411
32.5(40)
54.8(16)
250-400
50
19.4
0.8(44)
56
238
327
387
585
17.4
32.4
21.3
50.1
16.0
3181
33.8(24)
42.0(4)
500-1245
17
15.3
0.4
199
443
671
450
912
21.9
54.4
49.7
105.3
12.4
1190
30.1(14)
26.5(2)
20-120
12
13.9
1.2
44
92
69
287
335
6.6
14.6
12.5
29.4
15.1
3864
39
144(1)
December
130-245
19
11.5
1.1
72
119
190
301
420
17.6
25.3
31.2
57.1
15.4
5320(16)
53.3(13)
445(14)
- March
250-480
46
9.6(35)
0.8
63
260
338
265
537
10.6
22.8
19.9
41.2
16.9
2353(33)
24.5(30)
41.1(15)

500-1245
17
11.2
0.6
272
426
730
598
1035
69.1
93.4
44.4
137.4
12.5
2559
36.5(13)
78.4(5)
*A11 N and P concentrations mg/m3

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                    TABLE 3.  PHYSICAL, CHEMICAL AND BIOLOGICAL PARAMETERS,  ALL LAKE
                              SAMPLES USED IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                              RANGE OF INDICATED PARAMETER:  TOTAL NITROGEN  - mg/m3

Sample Period
Range of Values mg/m3
N
Water Temp. °C
Secchi Depth-m
NH3-N*
N02N03
Inorganic-N
Kj el-N
Total-N
P04-P
T-Soluble P
T-Particulate P
Total-P
TN/TP
i
Cell Density no. /ml
Chlor a_ Turner Units
Productivity mgC/m3/hr.
120-260
36
21.2
1.6
51
91
91.9
193
212
10.5
12.8
7.4
22.7
17.0
2206(30)
24(27)
20(8)
270-495
90
21.3
1.3
49
155
178
285
371
11.5
18.9
14.9
31.6
17.9
4234(62)
30(56)
36(21)
April - November
510-830
57
20.7
0.8
58
208
282
456
665
12.1
26.3
25.7
51.9
16.1
4347(42)
43(38)
51(14)
910-1580
17
19.7
0.5
166
310
506
654
983
22.5
50.0
52.1
103
15.8
8965
55(10)
83(5)
120-260
3
14.4
2.1
30
155
71.6
135
191
6.6
13.3
11.3
28.3
14.5
2534
16
23(1)
December
270-495
44
11.3
1.0
58
168
235
224
399
9.6
16.5
16.5
32.4
18.9
2940(39)
29(36)
45(17)
- March
510-830
35
9.6(24)
0.6
93
295
381
323
621
17.6
32.9
29.7
62.8
11.8
2360(23)
29(19)
32(12)

910-1580
12
11.5
0.6
296
372
714
837
1204
84.5
113
50.8
163
14.5
6579(10)
72(9)
114(5)
*A11 N and P concentrations mg/m3

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                    TABLE  4.   PHYSICAL,  CHEMICAL AND  BIOLOGICAL  PARAMETERS, ALL LAKE
                              SAMPLES  USED  IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                              RANGE OF INDICATED PARAMETER:   TOTAL  SOLUBLE-P - mg/m3

Sample Period
Range of Values mg/m3
N 	
Water Temp. °C
Secchi Depth-m
NH3-N*
N02N03
Inorganic-N
Kjel-N
Total-N
POA-P
T-Soluble P
T-Particulate P
Total-P
TN/TP
Cell Density no. /ml
Chlor a_ Turner Units
Productivity mgC/m3/hr.
April - November
5-19
102
22.0
1.4
50
89
146
290
378
6.4
8.4
13.1
22.7
21.2
4260(86)
30(77)
34(34)
20-40
57
19.3
0.9
77
193
269
395
577
10.4
25.5
25.1
51.5
13.6
3492(47)
39(39)
51(11)
45-80
17
17.7
0.6
105
347
408
453
682
31.0
60.1
41.8
100
8.1
9940(12
47(2)
149(12)
90-390
6
17.1
0.6
87
318
356
444
646
77.2
130
27.5
145
7.1
1327(4)
30(3)
-
5-19
42
11.8
1.0
56
191
260
236
440
5.3
10.4
13.6
24.0
21.6
2844(36)
28(37)
31(16)
December •
20-40
33
9.3
0.8
86
271
333
336
589
13.1
27.1
31.1
59.1
11.6
2793(25)
36(16)
50(16)
- March
45-80
9
12.2
0.5
204
336
540
435
111.
42.0
65.5
48.3
113
7.3
5107(7)
49(7)
179(1)

90-390
6
11.7
0.7
340
344
685
886
1230
187
213
52.5
265
5.1
5299
58
138(2)
*A11 N and P concentrations mg/m3

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                    TABLE 5.  PHYSICAL, CHEMICAL AND BIOLOGICAL PARAMETERS, ALL LAKE
                              SAMPLES USED IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                              RANGE OF INDICATED PARAMETER:  TOTAL-P - mg/m3

Sample Period
RanR_e of Values mg_/ma
N
Water Temp. °C
Secchi Depth-m
NH3-N*
Inorganic-N
Kjel-N
Total-N
P04-P
T-Soluble P
T-Particulate P
Total-P
TN/TP :
Cell Density no. '/ml
Chlor a. Turner Units
Productivity mgC/m3/hr.
5-18
53
22.2
1.7
38
69
122
224
298
5.6
7.4
4.5
11.8
24.7
2113(44)
24(41)
13(12)
April - November
20-40
81
22.1
1.1
60
108
188
382
470
8.0
14.7
13.6
27.5
17.8
5038(59)
32(49)
43(26)
43-80
36
19.5
0.8
60
194
269
422
575
13.8
28.9
31.4
60.2
11.1
5070(26)
50(23)
56(6)
85-410
27
17.5
0.5
113
341
428
489
725
34.6
63.2
52.6
118.0
8.3
7249(18)
44(17)
124(3)
5-18
16
10.7
1.3
58
179
249
180
375
4.9
7.7
5.3
12.5
29.5
1084(13)
18(13)
21(3)
December - March
20-40
35
10.9
0.9
71
197
285
280
485
6.5
14.8
13.9
28.2
18.1
3482(27)
31(27)
33(16)
43-80
27
11.4
0.6
82
260
326
333
589
13.7
28.0
28.9
57.8
10.4
3459(18)
37(14)
69(10)
85-410
17
10.5
0.6
241
342
584
588
930
82.4
109
62.6
172
6.1
4337
56(12)
77(6)
*A11 N and P concentrations mg/m3

-------
                                        TABLE 6.  PHYSICAL, CHEMICAL AND BIOLOGICAL PARAMETERS, ALL LAKE
                                                  SAMPLES USED IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                                                  RANGE OF INDICATED PARAMETER:  RATIO TN/TP

Sample Period
Range of Values ratio
N
Water Temp. °C
Secchi Depth-m
NH3-N*
N02N03
Inorganic-N
Kjel-N
Total-N
PO^-P
T-Soluble P
T-Particulate P
Total-P
TN/TP
Cell Density no. /ml
Chlor £ Turner Units
Productivity mgC/m3/hr
1.9-6.8
1*
20.5
0.8
73
164
271
486
648
34.7
61.9
55.5
113
5.1
12306(10)
72(8)
124(3)
April - November
7.0-10.8
42
20.1
0.7
71
240
301
364
545
16.1
36.9
35.6
64.6
8.5
3350(28)
39(27)
57(6)
11.0-15.7
46
20.2
1.2
61
122
202
302
402
11.3
21.0
13.5
36.5
13.5
3818(35)
28(32)
32(14)
15.0-21.5
44
21.9
1.3
55
84
152
357
435
7.2
11.7
12.5
24.3
17.9
4583(40)
36(33)
49(12)
22.0-87.0
50
21.9
1.4
52
123
187
360
465
8.0
9.3
6.4
19.4
30.6
3048(33)
24(27
22(13)
1.9-6.8
11
13.3
0.6
185
330
516
472
802
101
130
60.9
191
4.7
4963
56(9)
230(2)
December - March
7.0-10.8
25
10.1
0.6
144
243
367
357
609
17.2
34.4
37.5
70.7
8.7
4039
40(16)
44(4)
11.0-15.7
26
10.9
0.9
85
225
299
327
545
10.7
23.9
19.0
43.5
13.0
3266(20)
32(16)
26(8)
16.0-21.5
13
11.1
0.9
66
156
264
317
508
6.9
14.1
13.5
26.8
17.9
2669
32(11)
54(7)
22.0-87.0
18
10.5
1.2
79
263
335
238
499
5.1
9.4
7.2
16.6
32.7
1130(13)
19(14)
18(4)
*A11 N and P concentrations mg/m3

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                                          TABLE  7.  PHYSICAL, CHEMICAL AND BIOLOGICAL PARAMETERS, ALL LAKE
                                                   SAMPLES USED  IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                                                   RANGE OF INDICATED PARAMETER:  CELL DENSITY No./ml

Sample Period
Range of Values no. /ml
N_
Water Temp. °C
Secchi Depth-m
NH3-N*
Inorganic-N
Kjel-N
Total-N
PO^-P
T-Soluble P
T-Particulate P
Total-P
TN/TP
Cell Density no. /ml
Chlor £ Turner Units
1
Productivity mgC/m3/hr
April - November
62-599
23
15.8
0.8
133
232
345
339
559
14.9
32.9
32.0
64.2
14.9
410
21
-
604-1183
26
17.3
1.1
75
180
275
295
488
7.7
18.5
15.6
33.6
21.5
905
18(22)
15(7)
1210-2366
41
22.0
1.3
67
132
204
305
432
10.0
18.7
16.9
35.8
16.0
1801
33(37)
31(10)
2449-9926
46
22.9
1.2
55
92
147
354
447
9.1
15.9
18.0
33.9
17.5
4803
42(40)
44(18)
13085-82671
14
27.2
1.0
35
99
134
496
595
13.2
20.3
33.3
53.7
14.7
23326
67(11)
80(9)
62-599
22
10.9
1.0
75
224
299
311
535
17.6
28.6
22.4
51.1
16.8
305
20(18)
5(9)
December - March
604-1183
22
10.5
0.9
116
275
392
275
551
27.2
41.6
27.9
69.5
16.3
902
20(19)
27(9)
1210-2386
7
11.5
0.8
189
284
473
429
713
49.3
56.1
30.3
86.4
15.5
1712
31(5)
40(5)
2449-9926
23
11.5
0.9
74
221
295
318
540
18.0
27.0
22.0
49.0
13.9
5090
37
64(15)
11308-82671
7
11.8
0.7
151
93
203
786
806
29.1
45.4
79.3
144
8.4
25397
138
176
*A11 N and P concentrations mg/m3

-------
 nitrogen show a clearly indicated inverse relationship between cell number and
 nutrient.  However, the phosphorus mean values are less systematic in their
 relationship to cell number except that the overall values in the winter period
 seem to be somewhat higher than comparable values in the April-November growing
 season.

      A similar sorting for chlorophyll _a, Table 8, in four ranges of values,
 shows a strong positive relationship to cell density and productivity,  as would
 be expected, and to Kjeldahl nitrogen another direct measure of materials of
 biological origin.  Nevertheless, the phosphorus relationship continues to be
 ambiguous with higher mean values at the extreme ends of the ranges than found in
 the midranges.

      The sorting of the variable productivity, Table 9, describes,  for  those
 samples taken in the growing season, a parallel increase in organic nitrogen and
 soluble phosphorus with an increase in productivity.   The mean values for cell
 density and chlorophyll a_ show a similar parallelism.   Explanations for some of
 the anomalies may be explained in part by the fact that cell density may not be
 as precise a measure of biological production as cell volume which  is probably
 more accurately reflected in the measurements of chlorophyll a_ and  productivity.
 It might be noted here that the ratio between total nitrogen and total  phosphorus
 appears to be linked to biological activity.   In the  case of both chlorophyll  a_
 and productivity these parameters increased as this ratio systematically
 decreased.
 ALGAL ASSAY

 Growth in  NAAM

      One aspect  of  the  algal assay  that was  clearly  established by  the "inter-
 laboratory precision  test" was  the  degree variability between laboratories even
 though within  a  laboratory the  degree  of precision could be maintained at a
 reasonable level. **  One approach  to establishing  the reliability of the algal
 assay is the use of a growth control,  employing the  NAAM media, with each of the
 spiked, pre-treated samples.  This  is  essential to the determination of biomass
 grown in the spike samples which  is based on the  dry weight determination of
 biomass grown  in NAAM under the same conditions and  at the same time as that of
 the specific spiked series.  To provide some indication of the variability of the
 assay procedure  over the time span  of  several years  of this assay evaluation all
 of the samples grown in NAAM in conjunction  with  each of the specific lake
 samples have been arranged in Table 10.  This table  describes the actual dry
weight determination of biomass, maximum, minimum and mean values, standard
deviation, the unit absorbance  for  these same samples, and the mean values and
standard deviation for  unit absorbance.

      It is clear that over a period of several years variability is inherently
built  into a bioassay because of changes in  personnel which leads to small
changes in procedures in the assay  protocol, changes in purity of chemicals and
perhaps even changes in the growth  response  of the specific alga used in the
assay.  However, the biomass determinations, as systematically grown in NAAM do
indicate that on the average the values are  comparable over the several years.
The standard deviation of the sets  indicate  a variation on the order of 10-20% of

                                        20

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                    TABLE 8.  PHYSICAL, CHEMICAL AND BIOLOGICAL PARAMETERS, ALL LAKE
                              SAMPLES USED IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                              RANGE OF INDICATED PARAMETER:  CHLOROPHYLL a TURNER UNITS

Sample Period
Range of Values Units
N
Water Temp. °C
Secchi Depth-m
NH3-N*
N02N03
Inorganic-N
Kjel-N
Total-N
PO^-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Cell Density ho. /ml
Chlor a. Turner Units
Productivity mgC/m3/hr.

8-14
15
18.6
1.6
82
71
148
216
278
19.6
26.9
5.1
27.5
18.9
1164
11
7(5)
December
16-30
64
19.5
1.2
86
174
260
288
462
7.2
17.9
21.2
39.1
17.2
2030
23
26(13)
- March
31-60
40
23.5
1.1
59
111
174
333
445
9.1
15.3
17.2
33.3
16.4
5863
41
63(13)

64-139
12
25.8
0.7
36
148
183
617
765
19.5
43.4
51.6
95.0
10.0
14879
98
106(5)

8-14
17
10.1
1.2
94
270
364
215
486
16.7
24.2
15.9
40.1
21.3
644
12
9(6)
April -
16-30
24
12.0
0.8
104
248
352
309
556
28.1
38.4
23.9
62.4
14.8
1505
22
24(7)
November
31-60
19
11.7
0.9
102
240
342
353
593
23.7
40.5
19.0
59.6
17.2
4330
39
81(9)

64-139
6
11.7
0.6
160
162
322
738
900
54.0
77.1
58.3
135
8.0
18048
132
156(4)
*A11 N and P concentrations mg/m3

-------
      TABLE 9.  PHYSICAL, CHEMICAL AND BIOLOGICAL PARAMETERS,  ALL LAKE
                SAMPLES USED IN ALGAL ASSAYS, MEAN VALUES WITHIN VALUE
                RANGE OF INDICATED PARAMETER:  PRODUCTIVITY -  mgC/m3/hr

Sample Period April - November
Range of Values mgC/m3/h
N
Water Temp. °C
Seech i Depth-m
NH3-N*
N02N03
Inorganic-N
Kjel-N
Total-N
P04-P
T-Soluble P
T-Particulate P
Total-P
TN/TP
Cell Density no. /ml
Chlor a_ Turner Units
Productivity mgC/m3/hr
r. 1-27
17
23.3
2.1
42
56
100
285
343
6.1
8.9
5.1
13.9
25.2
2422
18(12)
14
30-57
19
23.9
1.2
46
95
167
359
472
7.2
13.6
16.3
29.2
15.5
9446(17)
36(14)
40
60-281
11
23.5
0.8
76
116
192
533
649
15.9
24.4
41.9
66.3
14.1
14124
71
95
December - March
1-27
15
9.0
1.1
85
166
251
249
415
9.0
16.3
16.4
32.7
16.9
1493
16(11)
13
30-57
8
9.7
1.0
41
214
256
249
463
6.2
14.5
21.5
36.0
14.9
4211
37
44
0-281
8
9.8
0.6
219
178
297
710
888
28.0
53.7
54.4
108
10.4
13314
110
150
*A11 N and P concentrations mg/m3
                                       22

-------
TABLE 10.  GROWTH OF SELENASTRUM CAPRICORNUTUM IN NAAM (14 DAYS, 24°C, 400 f.c.); MEAN VALUES OF
BIOMASS AND UNIT ABSORBANCE
Time Span of Number of NAAM Biomass In NAAMj mg/1 Unit Absorbance
Lake(s)
University,
Michie
Belews ,
Hyco
Kerr, Gaston,
Roanoke Rapids
Chowan River

Kerr Scott, Yadkin
High Rock, Tuckertown,
Badin, Tillery
Blewett Falls
NS
w James, Rhodhiss,
Hickory, Lookout Shoals
Norman, Mt. Island
Wylie, Fishing Creek
Wateree
Norman, Mt. Island
Wylie, South Fork
Wateree '
Mill Ponds and
Mattamuskeet, Phelps
S alters, Singletary
Waccamaw, White
Assays Samples Used Max. Min. Mean S.D. Max. Min. Mean S.D.
Oct. 12, 1971-
Sept. 11, 1973
Oct. 8, 1971-
Aug. 9, 1974
Sept. 20, 1971-
Jan. 7, 1975
Sept. 7, 1972-
Aug. 10, 1973
Dec. 6, 1971-
Jan. 7, 1975


Nov. 15, 1971-
Sept. 24, 1974
Nov. 15, 1971-
Sept. 24, 1974

Sept. 14, 1973-
June 25, 1974

Oct. 27, 1971
Jan. 21, 1975


13 148.8 110.9 125.0 10.6 307.1 225.9 259.1 23.5

19 144.3 88.4 119.3 12.6 403.1 211.1 263.7 45.8

20 136.9 85.1 114.9 13.1 275.1 182.0 240.2 33.9

4 131.8 117.5 125.1 6.1 262.0 232.1 247.5 14.5

7 123.2 92.2 107.7 13.8 259.4 207.6 230.2 17.4



8 135.9 67.8 112.1 20.4 367.9 178.8 252.7 53.1

7 113.4 101.8 119.2 11.6 367.9 235.0 268.6 46.3


16 123.2 83.9 110.0 10.0 309.0 200.9 266.7 28.3


6 114.1 85.1 99.7 11.1 367.9 182.0 242.5 65.0




-------
  the mean.  Growth of the test alga in medias or waters of low nutrient
  concentrations have shown greater variability.  It is such variation that must
  be kept in mind in evaluating the utility of a bio-assay.  In this particular
  study it has been instrumental in determining the approach used in the final
  evaluation of the results.

 Assay for Limiting Nutrients

      The procedure for establishing the nature and quantity of limiting nutrients
 in a surface water is to determine the growth response of a seeded sample, to the
 addition of phosphorus and nitrogen nutrient supplements or spikes, following
 pretreatment either by autoclaving or filtration.   For the summary of the
 extensive series of algal assays on the surface waters of North Carolina the
 growth response of spiked samples has been assembled in a series of tables which
 describe by the magnitude of the growth, under standard incubator conditions, as
 to whether growth was limited by phosphorus, nitrogen or both,  Tables 11-21.
 Since the mean growth of the control as well as standard deviation is indicated
 the significance of any specific response can be readily estimated.

      For purposes of this evaluation the biomass grown on addition of specific
 nutrient spikes were defined by the growth ranges  of 0-5, 5.1-15, 15.1-30 and
 over 30  mg/1.   This was growth in excess of that found in the lake control,  the
 biomass  grown in the unspiked sample.   The growth  of the lake control defines
 ambient  nutrient levels.   The data of these tables are clustered either by all
 samples  taken from one lake, samples taken from a  set of stations in a particular
 body of  water and compared to another set of stations in the same lake or in
 still other instances  where the number of samples  taken from any one lake was
 small, assay results from ponds that showed comparable limnological characteris-
 tics were  compiled into one set.   Biomass in the range 0-5 mg/1 was generally
 indicative of  no significant response particularly when examined in relationship
 to the variation indicated by the size of the standard deviation.   Growth at
 higher ranges,  5.1 mg/1 and above,  generally described a significant response to
 the  spike  and  was  indicative that the specific nutrient spike or combination  was
 probably limiting  to growth.

      From  the  growth response frequencies,  the number of samples found in each of
 the  growth  ranges,  a judgment can be made as to  whether the particular body of
 water was  algal  growth  limiting because of the inadequacy of the amount of
 phosphorus, nitrogen or both.   In reviewing  this information two factors  need to
 be kept  in mind.   For each  set  of samples the  number,  (N),  will  differ and it is
 this number which  defines  the total number  that  can  respond in  the  several growth
 ranges.  In addition the mean growth of the  lake control and  its standard
 deviation describe a baseline for referencing  the  magnitude of  growth  found in
 the various spiked samples.

 Control Growth at Nutrient Quantities  Following  Pretreatment  Procedures

     The data presented in Tables 11-21 provides the  opportunity to  examine two
aspects of the algal assay procedure.   One is  the  relationship of the  control
growth, nonspiked sample, to  the nutrient levels following  the two  pretreatment
procedures, autoclaving and filtration.   Does  the  growth  response of the  seeded
test alga indicate by some degree of correlation the magnitude of the  original


                                        24

-------
TABLE 11.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS

Pretreatment
Autoclaved
Lake Statlon(s) Mean Growth mg/1
Sample Period
University


July 1971 -
July 1973


Michie


July 1971 -
July 1973


Chowan
13, 17, CO 1,

Sept. 1972 -
Aug. 1973


Chowan
Al, RO 45

Sept. 1972 -
Aug. 1973


Spike Control
N=20 9.2
05P
SOP
75N
750N
5P+75N
50P+750N
N=17 4.4
05P
SOP
75N
750N
5P+75N
50P+750N
N=17 4.3
SW 1, 05P
SOP
75N
7 SON
' 5P+75N
I 50P+750N
N=8 2.3
05P
SOP
75N
7 SON
5P+75N
50P+750N
Spiked

8.2
11.1
8.0
10.5
9.2
27.9

5.1
9.5
4.2
5.7
5.5
23.9

5.6
7.2
7.9
8.6
8.2
26.9

2.5
3.5
3.3
3.9
4.6
24.0
S.D.
10.7
7.3
7.3
6.7
10.4
7.3
12.3
3.9
3.9
7.4
3.2
6.6
3.7
12.5
3.1
5.3
6.2
6.6
5.5
6.7
13.6
2.0
2.5
2.6
2.5
2.9
2.9
9.7
No. Responding, Growth Ranges
0-5 5.1-15

18
14
17
17
18
3

17
10
17
16
16
2

15
13
13
10
12
3

8
8
7
7
6
1

2
5
3
1
2
2

-
6
-
-
1
4

2
4
4
6
5
1

-
-
1
1
2
1
15.1-30 >30 mg/1

- -
1
-
2
-
13 2

-
1
-
1
-
7 4

-
-
-
1
-
8 5

-
- -
-
-
-
5 1
Mean Growth mg/1
Control Spiked
3.1
4.0
10.0
3.6
3.5
4.2
23.1
2.8
3.2
10.8
2.7
2.6
3.6
24.0
1.9
2.6
5.7
1.9
3.9
2.7
19.7
1.1
1.1
3.3
1.0
4.2
1.7
17.1
Filtered
No. Responding, Growth Ranges
S.D. 0-5 5.1-15
3.6
3.3
7.8
2.7
3.0
2.7
12.5
4.2
2.2
8.6
2.1
2.0
2.4
11.7
2.3
2.9
4.8
3.2
7.6
3.1
9.4
1.5
1.3
3.9
0.8
8.5
0.9
10.1

20
13
20
20
18
4

16
7
17
17
16
3

15
12
15
17
17
2

8
7
8
7
8
1

-
3
-
-
2
2

1
6
-
-
1
2

2
5
-
1
—
3

—
1
-
—
—
3
15.1-30

-
4
-
-
-
8

-
4
-
-
-
7

-
-
-
—
—
12

—
—
-
1
—
3
>30 mg/1

-
-
-
-
-
6

-
-
-
-
-
5

-
-
-
—
—
*~

—
—
—
—
—
1

-------
TABLE 12.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES,  CONTROLS AND ON ADDITION  OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Statlon(a)
Sample Period
Belews 1906

August 1971 -
April 1974



Belews 1116

August 1971 -
April 1974



Hyco 1906

October 1972 -
July 1974



Hyco 1116

October 1972 -
July 1974



Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N-15 1.8
05P
SOP
75N
750N
5P+75N
50P+750N
N-15 2.6
05P
SOP
75N
7 SON
5P+75N
50P+750N
N=9 5.1
05P
SOP
75N
750N
5P+75N
50P+750N
N-7 3.7
05P
SOP
75N
750N
5P+75N
50P+750N
Spiked

2.7
5.1
2.0
2.0
2.9
19.5

3.0
5.6
2.9
2.5
3.1
19.1

3.6
4.5
5.0
5.3
7.3
30.0

3.8
5.8
3.8
6.3
5.2
27.9
No. Responding, Growth Ranges
S.D. 0-5 5.1-15 15.1-30 >30 mg/1
1.8
1.8
3.9
1.8
2.2
2.0
10.6
1.7
1.9
3.5
1.7
1.9
1.6
9.7
3.2
3.3
3.0
3.6
4.5
5.6
7.5
2.4
2.9
4.5
3.0
8.8
3.2
13.4

15
10
15
15
15
1

15
11
15
15
15
2

9
9
9
6
7
-

7
6
7
6
7
1

_ - _
5 -
_
_
_
7 43

-
4 - -
_
_
_ _
3 82

_
_
_
3 - -
2 - -
1 62

_ _ _
1 - -
_
1
_
4 2
Mean Growth mg/1
Control Spiked
1.2
2.1
5.6
1.1
1.2
2.0
18.0
1.3
1.7
4.4
1.5
1.2
2.0
14.3
1.4
2.8
4.9
1.8
1.6
3.5
28.9
2.8
2.6
5.9
2.0
4.9
2.7
30.5
Filtered
No. Responding, Growth Ranges
S.D. 0-5 5.1-15
1.1
1.2
3.6
0.8
1.2
2.0
12.2
1.2
1.2
4.4
1.2
0.9
1.9
14.0
1.8
2.6
4.3
2.0
2.1
2.5
9.5
3.6
1.8
3.8
2.0
8.8
2.7
9.2

15
10
15
15
15
2

15
12
15
15
14
7

9
6
9
9
9
-

7
4
7
6
7
-

_
5
-
-
-
6

_
3
-
-
1
2

-
3
-
-
-
1

-
3
-
-
-
1
15.1-30 >30 mg/1

_ _
_ _
-
_
-
5 2

_
_ -
-
_
-
4 2

-
-
-
-
_
5 3

- -
-
-
I
-
3 3

-------
TABLE 13.
           GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
James
201, 206, 210, 212

July 1971 -
November 1973


Rhodhlss
3, 7, 9, 13, 1924,
1778, 1836

November 1971 -
November 1973

Hickory
1542, 1632, 1489

August 1971 -
November 1973
i

Lookout Shoals
1466, 1498, 1538

April 1972 -
November 1973


.Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N=7 2.0
05P
SOP
75N
750N
5P+75N
50P+750N
N=14 9.8
05P
SOP
75N
7 SON
5P+75N
50P+750N
N=5 9.9
05P
SOP
75N
750N
5P+75N
50P+750N
N=4 8.5
05P
SOP
75N
7 SON
5P+75N
50P+750N
Spiked

2.9
3.2
2.7
7.0
3.8
15.1

9.6
10.0
11.1
25.5
12.1
35.1

6.8
8.9
9.0
15.1
11.3
35.3

8.1
10.6
7.2
5.9
10.3
37.8
S.D.
2.2
2.2
1.8
3.1
11.3
2.3
10.8
6.2
6.4
6.1
7.2
18.8
6.2
14.1
6.2
4.2
3.1
5.6
12.9
5.7
4.8
9.5
10.5
7.0
9.2
7.3
9.6
7.7
No.
0-5

7
7
7
6
7
2

13
13
13
5
12
-

5
5
5
3
5
-

4
3"
4
4
3
-
Responding, Growth Ranges
5.1-15

-
-
_
_
-
2

1
1
1
1
2
2

-
_
-
1
_
-

-
1
-
-
1
-
15.1-30 >30 mg/1

-
-
_ _
1
-
3

-
-
_ _
6 2
_
9 3

-
_ _
-
1
-
5

-
_
-
-
_ _
2 2
Mean Growth mg/1
Control Spiked
0.6
1.1
1.3
0.9
3.9
1.9
9.5
5.4
6.4
7.8
6.8
13.6
7.0
28.9
0.8
1.2
5.9
0.4
0.4
1.9
21.1

2.8
3.6
9.3
3.3
3.3
4.6
S.D.
0.5
1.0
1.3
0.9
7.2
1.2
8.5
3.9
4.8
5.9
5.0
13.4
5.0
16.2
1.2
1.3
4.9
0.5
0.7
1.5
10.7

3.3
4.8
2.4
4.1
4.3
5.7
Filtered
No.
0-5

7
7
7
6
7
3

13
11
12
9
13
2

5
3
5
5
5
-

4
1
4
4
3
-
Responding, Growth Ranges
5.1-15

_
_
_
_
-
2

1
3
2
2
1
2

-
2
-
-
-
1

-
3
-
-
1
1
15.1-30 >30 mg/1

_ _
_ _
_ _
1
_
2

-
-
_
2 1
-
7 3

-
-
-
-
-
3 1

-
-
-
-
-
1 2

-------
TABLE 14.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
Norman
109, 116, 126, 1302
DC 26, RM 10

August 1971 -
November -197 3

Mt. Island
941, 960, 977

July 1973 -
January 1974


Wylle
789, 83, 831

July 1973 -
January 1974


Wylle
681, 708, 74, AC 22

July 1973 -
January 1974


Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N=ll 1.3
05P
SOP
75N
750N
5P+75N
50P+750N
N-12 0.4
05P
SOP
75N
7 SON
5P+75N
50P+750N
N=9 5.9
05P
SOP
75N
750N
5P+75N
50P+750N
N=13 5.8
05P
SOP
75N
750N
5P+75N
50P+750N
Spiked

2.8
6.5
1.4
1.2
3.2
19.3

1.3
4.1
1.1
1.3
3.7
15.9

4.5
4.9
5.5
13.2
6.2
22.2

5.7
5.5
5.8
9.1
6.7
21.6
No. Responding, Growth Ranges
S.D. 0-5 5.1-15 15.1-30 >30 mg/1
1.6
2.9
4.5
1.8
1.7
2.2
10.5
0.5
1.4
2.9
1.5
1.5
4.4
3.4
4.8
3.7
3.3
4.6
10.5
4.6
7.5
4.6
4.9
4.4
4.2
7.5
4.5
7.3

10
5
11
11
11
-

12
8
12
12
10
-

9
9
9
5
9
-

12
13
13
10
13
-

1 - -
6 - -
_ _
_
_
5 51

_
4 -
_
- -
2 - -
5 7 -

_ _
_
_
2 2 -
- — \ —
2 7 -

1 -
_ _ _
_
3 - -
_ _ _
7 6 -
Mean Growth rag/1
Control Spiked
0.5
1.1
6.4
0.7
0.7
1.3
18.3
0.1
0.6
4.5
0.5
0.3
2.5
14.5
1.0
2.2
3.9
1.6
4.1
2.5
18.6
2.3
3.0
4.7
2.7
4.2
3.5
17.6
Filtered
No. Responding, Growth Ranges
S.D. 0-5 5.1-15
1.2
1.4
3.7
1.1
1.2
1.3
9.5
0.1
1.0
2.6
0.7
0.4
4.7
6.0
2.0
2.3
2.2
3.0
8.1
2.3
7.3
3.8
4.1
4.5
3.9
5.8
4.0
9.0

11
3
11
11
11
1

12
7
12
12
11
1

8
8
9
7
9
1

13
11
13
12
12
1

_
8
_
-
_
4

-
5
-
-
-
5

1
1
-
-
-
1

-
2
-
1
1
7
15.1-30 >30 mg/1

_
-
_
-
_
5 1

-
_ _
-
-
1
6

-
-
-
1 1
- -
7

-
- -
-
- -
- -
5

-------
TABLE 15.
           GROWTH RESPONSE OF SELANASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
Wylie
SF 30

July. 197 3 -
January 1974


Fishing Creek
27, 31

August 1971 -
April 1972


Wateree
2, 58, 100, 104, 157

August 1971 -
January 1974

i
Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N=4 10.2
05P
SOP
75N
750N
5P+75N
50P+750N
N-3 28.9
05P
SOP
75N
7 SON
5P+75N
50P+750N
N-15 11.1
05P
SOP
75N
7 SON
5P+75N
50P+750N
Spiked

8.6
9.7
10.0
27.4
12.1
32.6

26.6
24.9
29.1
60.0
28.9
60.6

7.8
7.9
9.7
21.6
11.0
31.0
S.D.
4.3
3.9
3.2
5.4
8.1
5.7
6.4
4.6
8.7
6.4
7.8
8.5
6.9
6.5
9.5
7.5
7.3
8.1
15.9
8.1
9.8
No.
0-5

4
4
4
-
3
-

3
3
3
-
3
-

15
15
15
8
15
1
Responding, Growth Ranges
5.1-15

-
-
-
1
1
-

-
-
-
-
-
-

-
-
-
2
_
2
15.1-30 >30 mg/1

-
-
-
3
_
4

_
-
-
1 2
-
1 2

-
-
-
5
_ -
11 1
Mean Growth mg/1
Control Spiked
8.9
9.7
11.0
13.7
15.6
10.3
34.4
20.1
16.5
15.1
17.9
36.6
18.7
46.8
5.6
4.5
6.5
5.8
11.7
7.0
30.1
Filtered

No. Responding, Growth Ranges
S.D.
9.5
9.3
8.3
16.6
17.5
8.6
13.1
5.6
8.8
10.5
11.1
23.5
9.0
8.3
7.0
4.9
4.8
6.4
14.4
5.8
8.5
0-5 5.1-15

4
4
3
3
4
-

3
3
3
1
•3 _
-

15
12 3
14
10 2
15
1 1
15.1-30 >30 mg/1

_ _
_ _
1
1
_ _
4

_
_
-
2
_
3

_
-
1
3
- -
9 4

-------
TABLE 16.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
Kerr-Roanoke
19, 24

December 1971 -
March 1974


Kerr-Nutbush
118, 1308

January 1973 -
May 1974


Kerr-Nutbush
111, 114

January 1973 -
May 1974


Kerr-Nutbush
103, 108

September 1972
May 1974


Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N=8 12.1
05P
SOP
75N
7 SON
5P+75N
50P+750N
N-ll 12.4
05P
SOP
75N
750N
5P+75N
50P+750N
N=8 3.0
05P
SOP
75N
750N
5P+75N
50P+750N
N=13 2.0
05P
SOP
75N
7 SON
5P+75N
50P+750N
Spiked

8.9
8.6
12.2
26.4
12.7
33.5

10.8
13.2
12.9
32.7
13.0
35.9

3.3
5.9
3.6
8.6
5.5
28.9

1.7
6.0
2.3
4.3
3.5
25.9
S.D.
11.5
6.7
6.0
7.8
11.8
8.5
10.2
13.1
12.0
12.6
12.3
17.0
11.4
16.8
2.7
3.0
6.3
4.0
12.9
3.5
6.7
2.9
1.9
3.4
4.4
9.3
3.9
8.1
No.
0-5

8
8
8
2
8
-

10
9
10
2
9
1

8
6
8
6
7
-

13
9
12
11
13
-
Responding, Growth Ranges
5.1-15

-
-
-
2
-
3

1
2
1
1
2
1

-
2
-
1
1
-

-
4
1
1
-
1
15.1-30

-
-
_
3
-
4

-
-
-
6
-
6

-
-
-
-
-
6

-
-
-
1
-
9
>30 rag/1

-
_
_
1
-
1

-
_
-
2
-
3

-
-
-
1
-
2

-
-
-
-
-
3
Mean Growth mg/1
Control Spiked
4.2
5.3
7.2
5.0
13.7
6.5
28.3
4.7
4.0
4.3
5.6
14.6
5.6
22.6
0.7
8.0
1.8
0.9
4.6
1.0
19.9
1.6
2.0
7.3
1.7
5.3
3.6
23.8
S.D.
5.9
5.6
7.4
7.3
16.3
6.7
12.5
9.9
10.5
9.7
11.3
18.6
10.9
18.9
0.6
0.8
1.6
0.9
8.9
0.9
13.8
3.6
3.5
7.2
4.0
12.5
5.3
10.6
Filtered
No.
0-5

7
6
8
5
8
1

11
11
11
5
11
2

8
8
8
7
8
-

13
7
13
11
12
1
Responding, Growth Ranges
5.1-15

1
2
_
_
_
-

_
_
_
3
-
3

-
-
-
-
-
2

-
5
-
-
1
1
15.1-30 >30 mg/1

_ _
_ _
_ _
2 1
-
5 2

-
_ _
-
3
-
4 2

-
-
-
1
-
2 4

-
1
-
1 1
-
8 3

-------
TABLE 17.
           GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES,  CONTROLS  AND  ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
Kerr-Roanoke
1, 2, 8

July 1971 -
May 1974


Gaston
82, 166, 324

July 1971 -
March 1974


Roanoke Rapids
2, 28, 56

March 15, 1974

i

Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N-19 5.8
05P
SOP
75N
750N
5P+75N
50P+750N
N=5 7.5
05P
SOP
75N
750N
5P+75N
50P+750N
N=3 6.0
05P
SOP
75N
7 SON
5P+75N
50P+750N
Spiked

6.8
9.7
6.2
8.0
7.9
28.1

8.9
12.9
6.9
6.1
8.4
34.9

4.8
11.3
3.1
2.7
4.9
33.2
S.D.
6.0
6.8
7.2
6.4
9.6
6.7
12.6
4.9
5.2
5.5
5.0
4.3
4.7
14.6
2.3
3.4
3.3
2.7
2.9
2.7
1.4
No.
0-5

16
13
18
17
16
-

4
3
5
5
5
-

3
2
3
3
3
-~
Responding, Growth Ranges
5.1-15

3
5
1
1
3
2

1
2
-
-
-
1

-
1
-
-
-
—
15.1-30 >30 rng/1

-
1
-
1
-
14 3

-
-
-
-
-
2 2

-
-
-
-
-
3
Mean Growth mg/1
Control Spiked
1.4
2.
5.
1.
5.
2.
20.
1.2
1.
8.
1.
1.
1.
19.
0.2
0.
8.
0.
0.
0.
12.

4
6
8
6
4
8

9
5
4
3
8
1

5
2
1
1
3
4
S.D.
1.8
3.0
6.4
2.4
13.3
2.3
14.3
1.6
2.1
7.3
1.6
1.6
1.9
16.4
0.4
0.5
2.0
0.2
0.1
0.4
12.9
Filtered
No.
0-5

18
13
19
17
19
4

5
2
5
5
5
-

3
-
3
3
3
—
Responding, Growth Ranges
5.1-15 15.1-30 >30 mg/1

1 - -
4 2 -
_
- 2
- _ _
3 66

_
2 1 -
_
_
_
3 11

_
3 - -
_
_
- -
1 2 -

-------
TABLE 18.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
Kerr Scott
3 Stations

April 1974



High Rock
594, 654, 750

April 4, 1974



Tuckertown
516, 548, 580





Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N=6 3.0
05P
SOP
75N
7 SON
5P+75N
50P+750N
N-6 11.8
05P
SOP
75N
7 SON
5P+75N
50P+750N
N=6 16.1
05P
SOP
75N
750N
5P+75N
50P+750N
Spiked

3.8
5.1
1.9
2.5
3.1
20.0

7.9
9.5
10.7
25.4
13.7
35.5

16.4
20.7
14.9
15.8
15.0
37.3
S.D.
1.5
1.6
1.3
0.5
0.2
0.5
6.1
9.0
7.2
5.2
9.4
12.9
8.6
8.8
9.8
8.0
2.9
10.5
10.0
6.9
6.7
No.
0-5

6
6
6
6
6
-

6
4
4
-
4
-

6
4
6
6
6
-
Responding^ Growth Ranges
5.1-15 15.1-30 >30 mg/1

_
_
_
_ _
_ _
2 A -

- _ _
2 -
2
4 2 -
2 -
6 -

_ -
2 - -
_
_
_
6 -
Mean Growth mg/1
Control Spiked
0.3
0.4
2.2
0.3
0.5
0.6
4.7
4.0
6.1
15.3
6.4
6.2
4.9
16.0
0.1
0.1
15.0
0.1
0.3
0.8
13.1
S.D.
0.3
0.3
0.4
0.3
0.4
0.5
3.1
5.8
7.2
2.2
8.1
8.5
6.0
12.4
_
-
1.2
-
0.4
1.1
10.8
Filtered
No.
0-5

6
6
6
6
6
2

6
-
4
4
6
2

6
-
6
6
6
2
Responding, Growth Ranges
5.1-15 15.1-30 >30 mg/1

_
_
_ -
_
-
4 - -

_ - -
4 2 -
2
2 - -
-
4

- -
4 2 -
- -
- -
_
2 2

-------
TABLE 19.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM  IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Station(s)
Sample Period
Badin
438, 458, 490

April 5, 1974



Tillery

November 1971 -
April 1974



Blewett Falls
2, 26, 56

April 9, 1974


i
Pretreatment
Autoclaved
Mean Growth mg/1
Spike Control
N-6 9.8
05P
SOP
75N
750N
5P+75N
50P+750N
N-14 13.1
05P
SOP
75N
750N
5P+75N
50P+750N
N=6 10.7
05P
SOP
75N
7 SON
5P+75N
50P+750N
Spiked

12.5
18.7
13.3
16.2
11.0
37.8

14.2
19.6
13.0
14.5
14.4
30.3

14.4
18.5
13.6
13.5
13.0
33.3
S.D.
5.2
2.6
4.1
3.0
5.2
1.2
2.5
9.5
7.9
10.5
9.8
11.9
9.2
15.5
2.6
3.2
0.2
1.1
5.5
5.8
2.9
No.
O-.S

4
2
4
2
4


12
6
12
12
12
2

4
2
6
4
4
-
Responding, Growth Ranges
5. -15 15.1-30 >30 mg/1

2 -
2 2 -
2 -
4 -
2 -
42

2 -
8 - -
2 -
2 - -
2 - -
2 10 -

2 - -
4 - -
_
2 - -
2 - -
6
Mean Growth mg/1
Control Spiked
6.0
1.0
18.2
0.4
0.4
0.4
29.4
1.9
3.0
10.4
2.0
2.0
2.6
13.6
0.8
2.4
16.8
1.1
1.1
1.5
31.5
S.D.
0.4
3.2
1.5
0.4
0.6
0.6
3.0
1.6
1.2
9.5
1.7
1.7
1.3
13.5
1.0
1.8
2.2
1.5
1.1
1.4
1.8
Filtered
No.
0-5

6

6
6
6


12
8
14
14
14
8

6
—
6
6
6
-
Responding, Growth Ranges
5.1-15 15.1-3Q >30 mg/1

— — _
6 -

-
_
42

2 - -
2 4
_
_ _ _
_
4 2

_
2 4 -
_
_ _ _
_
2 4

-------
TABLE 20.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Lake Statlon(s)
Sample Period
lull, Davies,
McKensie, Orton





Jackson, Black






Finches, Hodgins
McNeils, Lytches
Maccamaw




Monroe, Jones (JO)
Salters, Singletary
White




Pretrear.ment
Autoclaved
Mean Growth mg/1
Spike Control
K- 4 0.6
05P
SOP
75N
750N
5P+75N
50P+750N
N= 3 21.8
05P
SOP
75N
750N
5P+75N
50P+750N
N- 7 2.9
05P
SOP
75N
7 SON
5P+75N
50P+750N
N=5 0.3
05P
SOP
75N
750N
5P+75N
50P+750N
Spiked

4.1
6.1
4.6
3.3
3.8
14.3

18.6
19.1
20.3
34.9
20.4
34.8

2.5
3.1
3.8
7.1
6.9
22.1

0.6
1.3
1.0
0.6
0.7
5.8
S.D.
1.2
7.4
9.2
7.0
4.2
3.9
3.8
3.9
5.7
5.4
6.1
8.3
6.9
8.0
2.7
1.9
2.4
3.8
7.5
9.8
12.2
0.4
0.4
1.1
1.6
0.4
0.5
8.6
No.
0-5

3
3
3
3
3
-

3
3
3
-
3
-

7
7
7
5
6
1

5
5
5
5
5
4
Responding, Growth Ranges
5.1-15 15.1-30 >30 mg/1

- 1 -
- 1 -
- 1 -
1
- 1 -
31-

_
_
_
3 - -
_
21-

- - -
- -
_
2 - -
1
222

- - -
- - -
_
_
_
1
Mean Growth mg/1
Control Spiked
0.2
0.6
1.0
1.5
0.4
0.8
13.0
16.6
14.4
15.8
16.5
24.8
16.8
28.9
1.2
1.9
3.3
1.3
2.2
5.3
16.1
0.3
0.5
0.9
0.4
0.5
0.5
2.0
S.D.
0.2
0.6
0.9
2.4
0.3
1.2
4.2
6.3
4.7
5.5
4.8
2.9
6.0
0.6
1.3
2.4
2.3
2.2
4.1
8.1
11.1
0.4
0.5
0.8
0.4
0.4
0.5
1.5
Filtered
No.
0-5

4
4
4
4
4
-

3
3
3
-
3
-

7
7
7
6
5
2

5
5
5
5
5
5
Responding, Growth Ranges
5.1-15 15.1-30 >30 mg/l

_ _ _
- _ _
_
_
_
22-

_
_
_
3 - -
_
21-

- - -
- - -
_
1 - -
11-
14-

- -
- - -
_
_
_
- - -

-------
TABLE 21.  GROWTH RESPONSE OF SELENASTRUM CAPRICORNUTUM IN LAKE SAMPLES, CONTROLS AND ON ADDITION OF
           NUTRIENT SPIKES OF NITROGEN AND PHOSPHORUS
Pretreatment Autoclaved
Lake, Station(s)
Sample Period
Jones (JP)






Johns






Crystal





I
Mattamuskeet, Phelps





Spike
N=l
05P
SOP
75N
750N
5P+75N
50P+750N
N=l
05P
50P
75N
7 SON
5P+75N
50P+750N
N=l
05P
SOP
75N
750N
5P+75N
50P+750N
N=2
05P
SOP
75N
7 SON
5P+75N
50P+750N
Mean Growth mg/1
Control Spiked S.D.
0.4
15.3
59.6
6.8
11.5
5.1
65.3
4.1
3.6
2.9
8.9
48.2
5.5
43.7
53.6
50.0
51.7
53.2
96.0
59.0
88.4
0.1
.45 .49
2.8 1.3
0.1
0.15 0.1
0.2 0.1
15.5 16.5
No. Responding, Growth Ranges
0-5 5.1-15 15.1-30 >30 mg/1

- - 1
- 1
- 1 -
- 1 -
1
- - - 1

1
1 _ _ —
1 - -
- - - 1
1
1
1
1 - -
1 - -
1 - - 1
1 - -
- 1
2 - -
2
2 - -
2 - -
2 - -
1-1
Mean Growth mg/1
Control Spiked
0.0
0.0
45.7
1.0
0.2
0.2
32.9
0.9
0.0
1.2
4.2
35.1
4.2
34.5
30.6
24.1
19.5
28.6
66.7
24.9
56.1
0.1
0.1
6.9
0.1
0.1
0.1
6.6
Filtered
No.
S.D. 0-5
_
1
_
1
1
1
-
-
1
1
1
-
1
-
1
1
1
-
1
-
2
3.8 1
2
-
-
6.6 1
Responding, Growth Ranges
5.1-15 15.1-30 >30 mg/1

- _
- - 1
_
_
_
- - 1

- _
_
_
- - 1
_
- - 1

_
_
- - 1
_
- 1 -

1
_
_
_
1 - -

-------
 nutrients.  The second is the relationship of the indicated nutrient limitation
 and the original raw water composition with respect to quantities of nitrogen and
 phosphorus.

      To examine the first relationship sets of samples from series of lakes or
 subsets of stations within lakes were assembled in Tables 22-25.   Each of the
 tables has been arranged according to a geographical or hydrologic pattern.  For
 example those lakes in Table 22 represents the series of impoundments on the
 Catawba River from Lake James downstream to Wateree Lake a distance of over 200
 river miles.  The data in the tables includes the mean control growth of the
 samples tested, their standard deviation for each of the pretreatment sets and
 correlation coefficients, significant at the 95% level, between the control
 growth and the quantities of soluble nutrients found in the sample after the
 indicated pretreatment.

      The use of autoclaving as a pretreatment with consequent solubilization
 produces higher nutrient levels and the inevitable greater control biomass.
 There was no instance in which the autoclaved growth was less than that of
 samples filtered as pretreatment although in one or two instances it was approx-
 imately the  same due to very marginal growth in both.   Only where the correlation
 was  significant at  95% or greater is the actual coefficient (r) value shown.
 Thus,  as shown in Table 22 for Lake James,  of the 7 samples tested when related
 to the nutrient levels, following pretreatment, no significant correlations were
 :ound.   However, in Lake Rhodhiss it is evident that j^ood correlations  were found
 ±n both autoclaved  and filtered samples with the phosphorus component of the
 water  and in the filtered sample with the quantity of  ammonia that was  present.
 In this series of impoundments of the Catawba River, it is apparent that the
 enrichment from municipal and industrial wastes entering Lake Wylie as  well as
 further downstream  results in stronger correlations as well as correlations with
 different  nutrient  species.   It would appear that the  correlation between biomass
 formed  by  the  seeded alga and the quantity  of nutrient improves as nutrient
 levels  increase,  probably due in part to improvement in precision of measurements.

     This  observation  was confirmed by the  assay data  derived from the  waters  of
 the two arms of  the John  H. Kerr Reservoir,  Table 23.   Strong correlations
 between control  growth and nutrients  were evident in the  section  of the lake
 noted as Kerr-Nutbush.   In downstream station sequence there  was  an indication of
 a  shift from good correlations with nitrogen and phosphorus,  Stations 118 and
 1308, to a solely phosphorus  correlated response at  Stations  108  and 103.   This
 paralleled the downstream decrease  in nutrient  concentrations.

     Occasionally,  such as the waters  from  Roanoke  Rapids,  Table  23 and Badin  and
 Tuckerton, Table 24, negative correlations were  found  which were  at the 95%
 significant level.  The meaning  of  these negative  relationships is not  clear.
 Omission of correlations  from these  tables may also mean  too  few  samples  were
 available for a correlation analysis.

Nitrogen and Phosphorus Limitation

     Using the data presented in Tables 11-21 an estimate was made as to  whether
 the particular set of waters was phosphorus  limited, nitrogen  limited,  or limited
by both nutrients.  This was determined for  autoclaved  and  filtered samples.   The


                                       36

-------
          TABLE 22.  CORRELATIONS OF SEEDED CONTROL GROWTH AND
                     NUTRIENT CONCENTRATIONS FOLLOWING PRE-TREATMENTS
Lake and Stations
James
201, 206, 210,
212
Rhodhiss
3, 7, 9, 13,
1724, 1778, 1836
Hickory
1542, 1632, 1689
Lookout Shoals
1466, 1498, 1538
Norman
109, 116, 126,
1302, DC26, RM10
Mt. Island
960, 977, 941
Wylie
83, 831, 789
681, 708, 74,
AC22
SF 30

Fishing Creek
27, 31
Wateree
2, 58, 100, 104,
157
Samples
Tested

7
14

5
4

11

12

9
13
4

3
15

Treat-
ment

A
F
A
F
A
F
A
F
A
F
A
F
A
F
A
F
A
F
A
F
A
F
Control
Growth*

2.0
0.6
9.8
5.4
9.9
0.8
8.4
2.8
1.3
0.6
0.4
0.1
5.9
1.0
5.8
2.3
10.2
8.9
28.9
20.1
11.1
5.7
S.D.

2.2
0.5
6.3
3.9
6.2
1.2
9.5
3.3
1.6
1.2
0.5
0.1
4.8
2.0
4.6
3.8
4.3
9.5
4.6
5.6
9.5
7.1
Corr. Coef. (r) P.>
NH-} NO?NO-} PO^-P

— — —
.7169
.5334
_
_ _ _
.9719
— _ _
. .
	 	 _
_ _ _
.9479 .6891
.7910
.7934 .8767
.7388
— _ _
.9335
_
.9177 .8359
.8012 .8454 .8004
.05
Total-P

~~
.6770
.6080
-
.9802
-
_
"
.6356
-
.9331
.7190
.8451
_
.9504
.9899
.6691
.6703
*Average of all samples, mg/1.
                                      37

-------
             TABLE 23.  CORRELATIONS OF SEEDED CONTROL GROWTH AND
                        NUTRIENT CONCENTRATIONS FOLLOWING PRE-TREATMENTS
 Lake and Stations
Samples   Treat-  Control          Corr.  Coef.  (r)  P>  .05
Tested	ment   Growth*  S.D.   NHg  N02NO-3 PO&-P Total-P
 Kerr-Roanoke
 19, 24,

 8, 2, 1
 Kerr-Nutbush
 118, 1308

 114, 11
   8
  19
  11
A
F

A
F

A
F

A
F
12.2
 4.2

 5.8
 1.4

12.4
 4.7

 3.0
 0.7
11.5
 5.9

 6.0
 1.8   -
                                              .6617   .7455
                                                           .5517
13.1 .7347 .7791
 9.9   -   .6049
                            2.7    -
                            0.6
           .8633
.6548  .6313

       .7833
.6579
108, 103

Gas ton
82, 166, 324
Roanoke Rapids

Chowan River
13, 17, C01, SW1
AL, R045

13

5

4

17

8

A
F
A
F
A
F
A
F
A
F
2.0
1.6
7.5
1.2
6.0
0.2
4.3
1.9
2.3
1.1
2.9 -
3.6 -
4.9 -
1.6
2.3 -
0.4 -
3.1 -
2.3 -
2.0 -
1.5 .7755
.7561 .7138
.9641 .9604
	 	
_ _
-.9566
- -
_ 	
- -
.7052
— —
*Average of all  samples, mg/1.
possibility, therefore, was one of three conditions of nutrient limitation
following two pretreatment procedures.  The results of all algal assays for all
bodies of water, or stations within a particular lake, are presented in a set of
tables for each specific nutrient limitation and pretreatment, Tables 26-31.  The
information assembled in these data sets includes mean control growth, the
correlation coefficients with mean nutrient levels in the treated samples, the
original mean lake quality with reference to inorganic nitrogen, P04-phosphorus,
soluble-phosphorus and total-phosphorus and the ratio of inorganic-N to soluble-P.
Thus in Table 26, 104 water samples taken from 11 different lakes or stations have
been shown to be phosphorus limited.  The control growth in these samples, mean
values, ranged from as low as 0.1 mg/1 to as high as 16.1 mg/1.  Where control
growth was reasonably high the correlation coefficient was positive to the
nutrient quantities in the pretreated water and showed in four instances to be
significant to total-P, in two instances to P04~P and one instance to N02+NO-J and
                                         38

-------
          TABLE 24.  CORRELATIONS OF SEEDED CONTROL GROWTH AND
                     NUTRIENT CONCENTRATIONS FOLLOWING PRE-TREATMENTS
Lake and  Stations
Samples  Treat-
Tested    ment
Control
Growth*  S.D.
        Corr. Coef. (r)  P^ .05
           NO?NOq  PO^-P  Total-P
Kerr Scott
2566,  2610,  2628

High Rock
594, 654, 750

Tuckertown
516, 548, 580

Bad in
438, 458, 490

Tillery
3, 11, 18, 22,
268, 300, 344

Blewett Falls
2, 26, 56
  14
           A
           F

           A
           F

           A
           F

           A
           F
           A
           F
  3.0
  0.3

 11.8
  4.0

 16.1
  0.1

  9.8
  0.5

 13.2
 10.7
  0.8
1.6
0.3

9.0
5.8

9.8
5.2
0.4

9.4
              .9294
.8910
      .8632
              .9607   .9017

.8411  .9075      -    -.8705



.9026  .8660   -.8660  -.8660

      .9519    .5699
2.6  .8130
1.0    -   .8537
              .9281   .8744
 Average of all samples, mg/1.
ammonia nitrogen.  In one case the correlation to ammonia was high but negative.

     Original lake quality showed a wide range of nutrient levels.  Inorganic
nitrogen was as low as 56 mg/m3 and as high as 1245 mg/m3.  Soluble phosphorus
ranged from 6 mg/m  to 84 mg/m3.  Following pretreatment by filtration, Table 27,
a considerably larger number of assayed samples were found to be phosphorus
limited, in this instance 191 from 21 different lakes or stations.  It is note-
worthy that fewer autoclaved samples showed phosphorus limitation than the same
samples following filtration pretreatment.  It might be concluded that auto-
claving solubilized sufficient quantity of phosphorus so that phosphorus
limitation for algal growth was masked.  Of the 191 samples that showed
phosphorus limitation following filtration pretreatment there was no clear
indication that the correlation between growth and nutrient levels in the
non-spiked waters was predominately to phosphorus or nitrogen.  The evidence
would appear to be evenly divided.

     The waters assayed to be nitrogen limited after autoclaving pretreatment
showed that of 132 samples from 17 sets of lakes or lake stations six were
highly correlated to the nitrogen quantity in the treated sample whereas nine
were positively phosphorus correlated and one negatively correlated to phosphorus
(Table 28).  Where the pretreatment was by filtration 69 assays in 10 sample
sets were nitrogen limited.  Of these four were correlated to the soluble
                                       39

-------
            TABLE 25.   CORRELATIONS OF SEEDED CONTROL GROWTH AND
                       NUTRIENT CONCENTRATIONS FOLLOWING PRE-TREATMENTS
Lake and Stations
Jackson, Black

Mattamuskeet ,
Phelps
Tull, Davies,
McKensie Orton
Finches, Hodgins
McNeils , Ly tches
Waccamaw
Monroe, Jones (JO)
Salters,
Singletary, White
Jones (JP)

Johns

Crystal

University

Michie

Belews 1906

1116

Hyco 1906

1116

Samples
Tested
3

2

4

7


5


1

1

1

20

17

15

16

9

7

Treat-
ment
A
F
A
F
A
F
A
F

A
F

A
F
A
F
A
F
A
F
A
F
A
F
A
F
A
F
A
F
Control
Growth*
21.8
16.6
0.1
0.1
0.6
0.2
2.9
1.2

0.3
0.3

0.4
0.2
4.1
0.9
53.6
30.6
9.2
3.1
4.4
2.8
1.8
1.2
2.6
1.2
5.1
1.4
3.7
2.8
S.D.
3.9
6.3
0.0
0.0
1.1
0.2
2.7
1.3

0.4
0.4

-
-
_
-
-
-
10.7
3.6
3.9
2.4
1.8
1.1
1.7
1.2 •
3.2
1.8
2.4
3.6
Corr. Coef. (r) P>L .)%
NH3 N09NO-} PO^-P Total-P
- - - -.9958
_ _ _
_ _ _ _
_ _ _ _
_ _ _ _
- - -
.9389
.7642 .8876

.9593
_

_ _ _
- - -
_ _ _ _
- - -
_ _ _ _
_ _ _ _
.7663 .7555
.7911
_ _ _ _
.7911
_ _ _ _
_
_ _ _ _
- - -
_ _ _ _
.7132 - .7173
- - - .8708
_ __ — . —
*Average of all samples, mg/1.
                                        40

-------
                            TABLE 26.  CORRELATION OF GROWTH IN NON-SPIKED SAMPLES OF PHOSPHORUS LIMITED WATERS,
AUTOCLAVED PRETREATMENT , WITH SAMPLE NUTRIENTS AND COMPARISON TO ORIGINAL
LAKE NUTRIENT CONCENTRATIONS
Lake or
Lake Station
Belews 1906
Belews 1116
Norman
Mt. Island
Gaston
Mlchie
Tuckertown
Tlllery
Blewett Falls
Jones (JP)
Mattamuskeet, Phelps
No. of Samples
Assayed
15
15
11
12
5
17
6
14
6
1
2
Control Corr. Coef. (r5_P>..05
Growth* NH-, N00NO, PO^-P Sol-P
1.8
2.6 - - - -
1.3
0.4 - - - .6356
7.5
4.4 - - - -
16.1 -.8411 .9075 - .8705
13.2 - - .6214 .5786
10.7 .8130 - .9281 .8744
0.4 - - - -
0.1
Original
Inorg-N
56
90
139
137
281
238
742
546
395
1245
97
Mean
Lake
Values
Quality mg/m3
PO^-P Sol-P
7
6
6
6
10
9
5
43
7
5
5
12
8
6
7
22
17
38
84
26
10
6

T-P
25
20
11
12
28
28
120
88
63
20
33
Ratio
Inorg-N/Sol-P
4.6
11.2
23.1
19.6
12.8
14.0
19.5
6.5
15.2
124.5
16.2
*Biomass, mg/1 14 days growth of J5. caprlcornutum in non-spiked water.

-------
                           TABLE 27.  CORRELATION OF GROWTH IN NON-SPIKED SAMPLES OF PHOSPHORUS LIMITED WATERS,
FILTERED PRETREATMENT, WITH SAMPLE NUTRIENTS AND COMPARISON TO ORIGINAL


Lake or No. of Samples
Lake Station Assayed
Belews 1906
1116
Hyco 1906
1116
Hickory
Lookout Shoals
Norman
Mt. Island
Kerr (1, 2, 8)
Gaston
Roanoke Rapids
University
MIchie
Chowan (13, 17, CO 1, SW 1)
Hich Rock
Tuckertovm
Bad in
Tillery
Blewett Falls
Jones (JP)
Mattamuskeet, Phelps
15
15
9
7
5
4
11
12
19
5
4
20
17
17
6
6
6
14
6
1
2
LAKE NUTRIENT
Control
Growth*
1.2
1.2
1.4
2.8
0.8
2.8
0.6
0.1
1.4
1.2
0.2
3.1
2.8
1.9
4.0
0.1
0.5
1.9
0.8
0.0
0.1
CONCENTRATIONS



Mean Values
Corr. Coef. (r) f>..05 Original Lake Quality mg/m3
NH^ N02NOi POi-P Sol-P Inorg-N
56
90
.7132 - .7173 156
111
177
.9719 - 177
- - 139
- - 137
.5517 - - 127
281
- - 303
.7911 - 144
.7911 - - 238
200
.9607 .9017 751
- - 742
-.9026 .8660 -.8660 -.8660 750
.9519 .5699 - 546
.8537 - - 395
1245
97
POA-P
7
6
7
6
a
8
6
6
7
10
5
10
9
15
11
5
20
43
7
5
5
Sol-P
12
8
11
10
13
13
6
7
16
22
19
14
17
36
60
38
40
84
26
10
6
T-P
25
20
19
19
38
38
11
12
20
28
34
35
28
80
115
120
102
88
63
20
33

Ratio
Inorg-N/Sol-P
4.6
11.2
14.2
11.1
13.6
13.6
23.1
19.6
7.9
12.8
15.9
10.3
14.0
5.6
12.5
19.5
18.7
6.5
15.2
124.5
16.2
*Biomass, mg/1 14 days growth of J3. capricornutum in non-spiked water.

-------
                            TABLE 28.   CORRELATION OF GROWTH IN NON-SPIKED SAMPLES OF NITROGEN LIMITED WATERS,
AUTOCLAVED PRETREATMENT, WITH SAMPLE NUTRIENTS AND COMPARISON TO
•ORIGINAL LAKE NUTRIENT CONCENTRATIONS
Lake or No. of Samples
Lake Station Assayed
Hyco 1906
Rhodhiss
Hickory
Wylie (789, 83, 831)
Wylie (681, 708, 74, AC22)
Wylie (SF 30)
Fishing Creek (27, 31)
Wateree (2,58,100,104,157)
Kerr (19, 24)
Kerr (118, 1308)
Chowan (13, 17, CO 1, SW 1)
(Al, Ro 45)
High Rock
Johns
Jackson, Black
Monroe, Jones (JO)
SIngletary, Salters, White
Crystal i
9
14
5
9
13
4
3
15
8
11
17
8
6
1
3
5

1
Mean Values
Control ' Corr. Coef . (r) P>.05 Original Lake Quality mg/m3
Growth* NH3
5.1
9.8
9.9
5.9
5.8
10.2
28.9
11.1
12.2
12.4 .7347
4.3
2.3
9.0
4.1
21.8
0.3

53.6
N09NOi PO^-P Sol-P Inorg-N
- - - 156
.7169 .6770 148
- - - 177
.9479 .6891 - 206
.7934 .8767 .7190 158
- - - 206
- - - 450
.9177 .8359 .6691 217
168
.7791 - - 142
- - - 200
.7052 - 130
.8632 - - 751
170
-.9958 752
.9593 - 57

900
P04-P
7
39
8
14
8
23
81
15
7
24
15
8
11
165
50
5

250
Sol-P
11
45
13
25
13
34
120
25
14
37
36
20
60
195
115
13

390
Ratio
T-P Inorg-N/Sol-P
19
69
38
31
21
78
135
49
35
108
80
42
115
250
170
17

410
14.2
3.3
13.6
8.2
12.1
6.1
3.7
8.7
12.0
3.8
5.6
6.5
12.5
0.9
6.5
4.3

2.3
*Biomass, mg/1 14 days growth of J3. capricornutum in non-spiked water.

-------
                            TABLE 29.   CORRELATION OF GROWTH IN NON-SPIKED SAMPLES OF NITROGEN LIMITED WATERS
FILTERED PRETREATMENT , WITH SAMPLE NUTRIENTS AND COMPARISON TO ORIGINAL
LAKE NUTRIENT CONCENTRATIONS
Lake or No. of Samples
Lake Station Assayed
Rhodhiss
Wylie (789, 83, 831)
Wylie (SF 30)
Fishing Creek
Wateree
Kerr (19, 24)
Kerr (118, -1308)
Johns
Jackson, Black
Crystal
14
9
4
3
15
8
11
1
3
1
Control
Growth*
5.4
1.0
8.9
20.1
5.7
4.2
4.7
0.9
16.6
30.6
Corr. Coef. (r) P> .05
NHi N02N03 POA-P Sol-P
.5334 - - .6080
.7910 .9331
.9335 .9504
.9899
(Pi. 10)
.8012 .8454 .8004 .6703
.6617 .7455
.6049 .6548 .6313
-
-
-
Mean Values
Original Lake Quality ms/m3
Inorg-N
148
206
206
450
217
168
142
170
752
900
P04-P
39
14
23
81
15
7
24
165
50
250
Sol-P
45
25
34
120
25
14
37
195
115
390
Ratio
T-P Inorg-N/Sol-P
69
31
78
135
49
35
108
250
170
410
3.3
8.2
6.1
3.7
8.7
12.0
3.8
0.9
6.5
2.3
*Biomass, tng/1 14 days growth of J^. capricornutum in non-spiked water.

-------
                            TABLE 30.   CORRELATION OF GROWTH IN NON-SPIKED SAMPLES OF PHOSPHORUS & NITROGEN LIMITED WATERS,
AUTOCLAVED PRETREATMENT , WITH SAMPLE NUTRIENTS AND COMPARISON TO ORIGINAL
LAKE


NUTRIENT CONCENTRATIONS
Lake or
Lake Station
Hyco 1116
James
Lookout Shoals
Kerr (111, 114)
Kerr (103, 108)
Kerr (1, 2, 8)
Roanoke Rapids (2,
University
Kerr Scott
Badin
Tull, Davies, Orton,
Finches, Hodgins ,
Lytches, Waccamaw
No. of Samples
Assayed
7
7
4
8
13
19
28, 56) 4
20
6
6
McKensie 4
McNeils 7

Control
Growth*
3.7
2.0
8.4
3.0
2.0
5.8
6.0
9.2
3.0
9.8
0.6
2.9

Corr. Coef. (r) P .05
NH-i NO?NOi POi-P Sol-P
.8708
- - -
- - .9802
.8633 - .7833
.7561 .7138
_
- - -.9566
.7663 .7555
.9294
- - -
_
.9389

Mean Values
Original Lake Quality mg/m3
Inorg-N
111
87
177
80
79
127
303
144
282
750
334
253

PO,,
6
6
8
7
7
7
5
10
5
20
63
15

-P Sol-P
10
7
13
7
9
16
19
14
22
40
70
28

T-P
19
18
38
23
19
20
34
35
47
102
105
42

Ratio
Inorg-N/Sol-P
11.1
12.4
13.6
11.4
8.8
7.9
15.9
10.3
12.8
18.7
4.8
9.0

*Biomass, rog/1 14 days growth j^. capricornutum in non-spiked water.

-------
                            TABLE 31.   CORRELATION OF GROWTH IN NON-SPIKED SAMPLES OF PHOSPHORUS & NITROGEN LIMITED WATERS,
FILTERED TREATMENT, WITH SAMPLE NUTRIENTS AND COMPARISON TO ORIGINAL LAKE NUTRIENT


Lake or No. of Samples
Lake Station Assayed
James
Wylie (681, 708, 74, AC 22)
Kerr (111, 114)
Kerr (103, 108)
Chowan (Al, RO 45)
Kerr Scott
lull, Davies, Orton.McKensie
Finches, Hodgins, McNeils
Lytches, Waccamaw
Monroe, Jones (JO)
Singletary, Salters, White
7
13
8
13
8
6
4
7
5
CONCENTRATIONS

Control Corr. Coef. (r) P^.05
Growth" NH,
0.6
2.3
0.7
1.6
1.1 .7755
0.3 .8910
0.2
1.2
0.3
N02NO-} P04-P Sol-P
_ _ _
.7388 .8451
.6579
.9641 .9604
_
- - -
-
.7642 .8876





Mean Values
Original Lake Quality mg/m3
Inorg-N
87
158
80
79
130
282
334
253
57
PO.-P
6
8
7
7
8
5
63
15
5
Sol-P
7
13
7
9
20
22
70
28
13
T-P
18
21
23
19
42
47
105
42
17

Ratio
Inorg-N/Sol-P
12.4
12.1
11.4
8.8
6.5
12.8
4.8
9.0
4.4
*Biomass, mg/1 14 days growth of jS. capricornutum  in non-spiked water.

-------
nitrogen component of the filtered sample and 12 were highly correlated at a
significant level to the soluble phosphorus components,  (Table 29)  of twelve
autoclaved sampling sets, one correlated to both nitrogen and phosphorus,  nine
were correlated to soluble phosphorus while only one showed a significant  corre-
lation to soluble nitrogen and one was negatively correlated to soluble phosphorus
(Table 30).  In the samples pretreated by filtration 71 in nine sets were  phos-
phorus and nitrogen limited with three correlated to the soluble nitrogen  and
six to the soluble phosphorus components (Table 31).

     These results can be summarized in two ways.  In one, shown in Table  32,
the mean control growth of each of the set of samples that were phosphorus,
nitrogen or phosphorus and nitrogen limited following the two pretreatment
procedures has been correlated to the original lake nutrient quality,
specifically the soluble nitrogen and phosphorus components and the ratio  of
these two.  In this presentation the actual (r) value derived from the
correlation determinations xs shown without noting a level of significance.  The
r values which are less than .5 are generally assumed to be of little or no
significance.  It is evident that autoclaving as a pretreatment resulted in


TABLE 32.  RELATIONSHIP OF CONTROL GROWTH, SEEDED SELENASTRUM CAPRICORNUTUM IN
           AUTOCLAVED AND FILTERED WATERS AND NUTRIENT LEVELS OF ORIGINAL  RAW
           WATER

                                        	Corr. Coef. (r)	
Nutrient Limitation                                                      Ratio
Based on Response to Spikes	N	Inorg-N	Soluble P	N/P

       Phosphorus

           Autoclaved            11      .3151           .7928           -.2992

           Filtered              45     -.3844          -.2380           -.3565

       Nitrogen

           Autoclaved            17      .7653           .8270           -.2932

           Filtered              10      .9103           .7439            .4720

  Phosphorus and Nitrogen

           Autoclaved            12      .4172          -.1344           -.0067

           Filtered               9     -.2331          -.3693            .3042
control growths in the phosphorus limited series that was highly correlated to
the original soluble phosphorus level, an r value of .7928.  All other
correlations were at nonsignificant levels.  In contrast the nitrogen limited
samples following both autoclaved and filtered pretreatment showed unusually
                                      47

-------
 high correlations to both the inorganic nitrogen as well as  soluble  phosphorus
 of the original lake quality.  Samples linn'ted in both phosphorus  and nitrogen,
 whether autoclaved or filtered as pretreatment,  showed correlations  with
 neither of the soluble nitrogen or phosphorus components of  the original  lake
 water.   It would thus appear that this particular analysis did  not discriminate
 in any consistent manner between the two pretreatment procedures to  indicate
 whether one or the other was preferable in relating the response of  the algal
 assay growth to the quality of the original  lake water.

 Nutrient Limitation and Original Lake Quality

      In the second overall summary,  Table 33,  the mean value for all autoclaved
 and filtered samples for each of the limitation  series,  phosphorus,  nitrogen,
 and phosphorus and nitrogen are compared to  the  mean values  of  the original lake
 water nutrient concentrations.   A clear growth limitation relationship emerges,
 regardless  of the limiting nutrient,  in the  autoclaved pretreated  samples but
 not in  the  filtered samples.  This would be  expected.  It also shows that those
 samples  that  were  nitrogen limited, whether  autoclaved or filtered, had an
 average  growth which was  greater than  either the phosphorus limited or
 phosphorus  and nitrogen limited  samples.  When these  summary data are compared
 to  the mean values  of the  original lake water nutrient levels it is clear that
 the phosphorus limited samples had at  least  a soluble phosphorus component of
 about 23 mg/m^ with  a mean  ratio of nitrogen/phosphorus of about 14.   Nitrogen
 limited  samples averaged considerably higher in mean  soluble phosphorus,  3-5
 times, depending on whether the  sample was autoclaved or filtered but the
nitrogen/phosphorus ratio was lower in the range of 5-7 for the two pretreat-
ment procedures.  Phosphorus and nitrogen limited samples showed soluble
phosphorus  about the same as for  the phosphorus limited but nitrogen was
considerably  the lowest on  the average and the ratio of soluble nitrogen to
soluble phosphorus was in  the range of 9-11.
                                    48

-------
                           TABLE 33.  NUTRIENT LIMITATION OF ALGAL ASSAYED SAMPLES,
                                      CONTROL BIOMASS AND ORIGINAL LAKE NUTRIENT QUALITY
Nutrient Pre- No. of Mean Control
Limitation Treatment Assays Biomass, mg/1
Phosphorus Autoclaved 104

Filtered 191

Nitrogen Autoclaved 132
Filtered 69
Phosphorus and Autoclaved 105
Nitrogen
Filtered 71
5.31
(5.81)
1.37
(1.37)
12.15
9.81
4.70
0.92
Mean Values
Original Lake Nutrients
Inorg-N
360
(272)
326
(281)
293
335
227
162
P04-P
9.9
(10.4)
9.8
(10.1)
42.9
66.8
13.3
13.8
Sol-P
21.4
(22.6)
22.3
(22.9)
68.5
100.0
21.3
21.0
mg/m3
T-P
40.7
(42.8)
45.1
(46.4)
98.1
133.5
41.8
37.1
Mean Ratios
Inorg-N/Sol-P
24.3
(14.3)
18.6
(13.3)
7.3
5.5
11.4
9.1
(    )  mean values with Jones Pond (JP) deleted because of unusual nitrogen concentration.

-------
                                  SECTION VIII

                                   REFERENCES


 1.   Provisional Algal Assay Procedure.   Joint Industry/Government Task Force on
     Eutrophication.   New York,  62 p.,  February 1969.

 2.   Algal Assay Procedure;  Bottle Test.   National Eutrophication Research
     Program,  Environmental Protection  Agency, Corvallis Oregon.   August 1971.
     82 p.

 3.   Maloney,  T. E.  and W.  E.  Miller.   Algal Assays:   Development and Application.
     Special Technical Publication 573.   Am. Society  for Testing  Materials,
     Philadelphia,  p.  344-355, 1975.

 4.   Weiss, C. M.,  and R. W. Helms.  The  Interlaboratory Precision Test, An
     Eight Laboratory Evaluation of the Provisional Algal Assay Procedure Bottle
     Test.  National Eutrophication Research Program,  Environmental Protection
     Agency, Corvallis, Oregon,  70 p.,  October 1971.

 5.   Sturm, R. N. and A. G.  Payne.  Environmental Testing of Trisodium Nitrilotri-
     acetate:   Bioassays for Aquatic Safety and Algal  Stimulation.  In:  Bioassay
     Techniques and Environmental Chemistry, Glass, G. E. (ed.).   Ann Arbor,
     Ann Arbor Science Publishers, Inc.,  p. 403-424,  1973.

 6.   Payne, A. G.  Environmental Testing  of Citrate:   Bioassays for Algal
     Stimulation.  In:  Proceedings 16th  Conference Great Lakes Research.
     International Association Great Lakes Research,  p. 100-115,  1973.

 7.   Mitchell, D. and J. C.  Buzzell,  Jr.   Estimating  Eutrophic Potential of
     Pollutants.  Jour. Sanitary Engineering Division, Proceedings American
     Society Civil Engineers,  97 (SA4):453-465, 1971.

 8.   Toerien,  D. F.  and D.  J.  Steyn.  Application of  Algal Bioassays in Eutrophi-
     cation Analyses.   South African Jour. Science. 69:79-82, 1973.

 9.   Francisco, D.  F.  and C. M.  Weiss.  Algal Response to Detergent Phosphate
     Levels.  Jour.  Water Pollution Control Federation.  45:481-489, 1973.

10.   Steyn, D. J.,  D.  F. Toerien and J. H. Visser. Continuous Culture Algal
     Bioassays.  South African Jour.  Science.  70:277-278, '1974.

11.   Miller, W. E.,T.  E. Maloney and J. C. Greene. Algal Productivity in 49 Lake
     Waters as Determined by Algal Assays.  Water Research.  8:677-679, 1974.

12.   Payne, A. G.  Responses of the Three Test Algae  of the Algal Assay Procedure:
     Bottle Test.  Water Research.  9:437-455, 1975.
                                        50

-------
13.  Doemel, W. N. and A. E. Brooks.  Detergent Phosphorus and Algal Growth.
     Water Research.  9:713-719, 1975.

14.  Greene, J. C., W. E. Miller, T. Shiroyama and T. E.  Maloney.   Utilization
     of Algal Assays to Assess the Effects of Municipal,  Industrial, and
     Agricultural Wastewater Effluents Upon Phytoplankton Production in the
     Snake River Sytem.  Water, Air, and Soil Pollution.  4:415-434, 1975.

15.  Weiss, C. M., T. P. Anderson and D. R. Lenat.   Environmental  Assessment,
     Belews Creek-Belews Lake, North Carolina, May 1971 - June 1972, Year  II.
     Department of Environmental Sciences and Engineering, University of North
     Carolina at Chapel Hill, Chapel Hill, N. C.  October 1972,  i-x, 232 p.

16.  Weiss, C. M., T. P. Anderson, P. H. Campbell,  D. R.  Lenat,  J. H.  Moore and
     S. L. Pfaender.  Environmental Comparison, Belews Lake - Year III and Lake
     Hyco, North Carolina, July 1972-June 1973.  Department of Environmental
     Sciences and Engineering, University of North Carolina at Chapel Hill,
     Chapel Hill, N. C.  April 1974, i-xxvii, 510 p.

17   Weiss, C. M., P. H. Campbell, T. P. Anderson,  S. L.  Pfaender.  The Lower
     Catawba Lakes - Characterization of Phyto- and Zooplankton Communities
     and Their Relationships to Environmental Factors. Department of Environ-
     mental Sciences and Engineering, University of North Carolina at Chapel
     Hill, Chapel Hill, N. C.  January 1975, i-xxii,  396  p.

18.  Weiss, C. M. and J. H. Moore.  The John H. Kerr Reservoir,  Virginia-
     North Carolina, A report for the OECD North American Project  Defining Its
     Limnological Characteristics, Productivity, Nutrient Budgets  and
     Associated Parameters.  The Department of Environmental Sciences and
     Engineering, School of Public Helath, University of  North Carolina at
     Chapel Hill, Chapel Hill, N.C.  May 1975, i-v, 39 p.
                                     51

-------
APPENDIX A.  SEASONAL MEAN VALUES OF LIMNOLOGICAL PARAMETERS, LAKE SAMPLES
             USED IN ALGAL ASSAYS
Lake
Season
No. of Samples
Water Temp. °C
Secchi Depth-m
NH--N

Inorg-w
Kjel-N
Total-N
PO -P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a Turner Units
Productivity mgC/m3/hr.
(9) Belews 1116
Apr. -Nov.
9
21.9
2.0
20
36
56
277
313
6.7
11.7
13.5
24.6
20.5
4237(7)
14(4)
13(5)
Dec. -March
5
8.4
1.9
131
98
226
360
458
6.6
8.3
11.0
19.6
25.9
698
10(1)
12(2)
(11) Universiix 	
No. of Samples
Water Temp. °C
Secchi Depth-m
NH--N
NO^+NO.-N
Ino rg-N
Kjel-N
Total-N
PO -P
T-Soluble P
T-ParCiculate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a. Turner Units
Productivity mgC/m3/hr.
12
23.3
1.0
86
58
144
541
600
10.2
14.4
21.7
35.4
22.0
3980(8)
-
44(7)
7
8.4
0.5
106
297
403
510
820
17.1
32.1
35.0
67.1
10.6
906(4)
—
25(5)
(9) Belews
1906 (lO).Hvco 1116 fin) Hvco ;906
Apr. -Nov. Dec. -March Apr. -Nov. Dec. -March Apr. -Nov.
9
18.9
2.3
38
52
90
296
348
6.2
7.7
13.6
20.4
28.8
1435(6)
14(4)
9(5)
r-m MI
12
23.3
1.0
60
178
238
540
7.18
9.0
17.3
10.9
27.5
27.5
7531(8)
~
22(6)
5 ,5 2 6
8.6 20.5 11.6 22.6
2.1 0.7 0.7 1.5
125 44 55 56
80 67 115 100
205 111 170 156
220 416 225 193
300 483 340 293
54 6.0 7.5 6.6
11.6 10.2 15.0 10.8
1.0 9.0 250 9.8
13.6 19.2 40.0 18.8
23.8 28.4 8.6 16.0
734 1564 2571 2058
9(1) 18(2) 23 19(4)
9(2) 21(3) 23 18(4)
rHp _ ..
5
7.9
0.4
110
398
508
397
795
14.0
24.0
47.0
71.0
14.9
496
™"
5
Footnotes

Number ( ) preceeding lake name is
used in Table 1 and Figure 1.
Number (s) following lake name are
on the specific lake.
Number ( ) following a mean value
samples averaged for that value
from number at top of column.
All N and P concentrations mg/m3.






Dec. -March
2
13.5
0.7
45
128
173
185
313
7.5
15.0
70.0
85.0
6.2
2029
23
18



identification

sampling stations

is number of
if different








                                                                                         (Continued)

-------
APPENDIX A.
SEASONAL MEAN VALUES OF LIMNOLOGICAL PARAMETERS, LAKE SAMPLES
USED IN ALGAL ASSAYS Continued
' (15.) Hickory
Lake
Season
No. of Samples
Water Temp. °C
Secchi Depth-m
NH.-N
3
NO +NO — N
Inorg-n
Kjel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor £ Turner Units
Productivity mgC/m3/hr.
(13)
Apr. -Nov.
6
15.7
1.8
28
59
87
205
256
6.2
7.0
12.0
17.5
21.4
1620(3)
20.6(3)
-
James
Dec. -March
None
-
-
-
-
-
-
-
_
_
-
-
_
-
-
-
(19) Hylie 789, 831
No. of Samples
Water Temp. °C
Secchi Depth-m
NH.-N

Ino rg-N
Kjel-N
Total-N
PO . -P
T-Soluble P I
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a Turner Units
Productivity mgC/m3/hr.
7
23.3
1.0
76
130
206
250
380
13.6
25.1
6.3
31.4
13.5
1854(6)
26(6)
-
1
15.1
0.8
140
395
535
220
615
30.0
24.0
31.0
65.0
9.4
621
9
-
(14.)
Apr. -Nov.
5
14.4
0.9
120
73
148
350
401
38.6
45.0
26.7
68.7
10.6
2033(2)
20(2)
-
(19) Wylie
9
23.3
1.5
74
84
158
267
351
8.1
13.2
7.9
21.1
16.7
1849
28
-
Rhodhlaa
Dec. -March
None
-
-
-
-
-
-
-
_
-
-
-
-
-
-
-
681. 708. AC22
2
12.3
0.6
85
390
475
205
595
22.5
31.0
34.0
65.0
9.1
341
15
-
(16) Lookout
Shoals
Apr. -Nov. Dec. -March
9
15.8
1.1
62
115
177
296
387
7.5
12.5
25.0
38.3
16.6
5014(4)
28(4)
-
(19) Wvlie
3
25.1
0.7
58
148
206
380
528
23.3
33.6
44.6
78.3
6.7
3272
102
—
8
15.8
1.1
51
114
166
327
411
7.9
13.6
24.3
38.7
18.6
5678(3)
30(3)
-
SF 30
1
13.2
0.6
180
420
600
250
670
50
60
45
105
6.3
823
15
"•
(17)
Apr. -Nov.
15
22.1
1.7
58
81
139
215
297
5.5
6.4
3.9
10.5
29.5
2267(13)
27(13)
-
Norman
Dec. -March
5
9.1
1.1
37
309
346
156
465
5.0
6.6
7.8
14.4
33.1
841
15
-
(20) Fishine Creek
3
21.7
0.8
83
366
450
500
867
81
120
30
135
6.4
—
—
None
-
-
-
-
-
-
-
-
-
-
—
-
—
—
(tftl
Apr.-Nov
12
24.1
1.2
35
102
137
231
334
6.0
6.5
5.3
12.2
27.6
1348(9)
27(9)
—
C2T)
11
23.5
0.9
88
129
217
419
548
14.8
25.4
25.6
48.9
13.4
11,112(9)
54(9)
™
Mt, Island
Dec . -March
3
10.1
0.9
28
320
348
113
433
5.0
6.6
6.6
13.3
32.6
945
17
—
Unt-prpp
3
11.3
0.4
220
441
661
350
741
45.0
70.0
45.0
115
6.9
795
15
~
                                                                                                 (Continued)

-------
APPENDIX A.
             SEASONAL MEAN VALUES OF LIMNOLOGICAL PARAMETERS, LAKE SAMPLES
             USED IN ALGAL ASSAYS Continued
Lake
Season
No, of Samples
Water Temp. °C
Secchi Depth-m
NH,-N
NO^+NO -N
Inorg-N
Kjel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a. Turner Units
Productivity mgC/m3/hr.

No. of Samples
Water Temp. °C
Secchi Depth-m
NH,-N
NO +NO -N
Inorg-N
Kjel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor ji Turner Units
Productivity mgC/m3/hr.
(221 Kerr Scott
Apr. -Nov. Dec. -March
3 None
13.7
0.8
62
220
282
200
420
5.0
21.6
25.0
46.6
9.3
399
19
-
(26) Tillerv
3
15.4
0.3
45.8
501
546
337
838
42.5
84.2
36.3
88.0
7.4
1015
21.3
-
(211 H-tph Rnrk
Apr. -Nov. "Dec. -March
3
14.9
0.3
248
503
751
343
846
11.0
60.0
55.0
115
7.4
426
26.3
-
(271 Biewpft: Falls
3
13.5
0.3
15.3
380
395
206
670
7.0
26.0
37.3
63.3
10.6
921
17.6
-
(241 Turlcprt-nn
Apr. -Nov. Dec. -March
3
14.0
0.2
242
500
742
470
970
5.0
38.3
81.7
120
8.1
559
28.7
-

















C?"5) l
Apr. -Nov.
2
13.7
0.3
245
505
750
345
850
20.0
40.0
62.5
102
8.8
1929
29.0
-

















^Hln
Dec. -March

-
-
_
_
-
-
-
„
_
-
-
-
-
-
-

















                                                                                  (Continued)

-------
APPENDIX A.
SEASONAL MEAN VALUES OF LIMNOLOGICAL PARAMETERS, LAKE SAMPLES
USED IN ALGAL ASSAYS Continued
Lake John.H. Kerr 19, 20, 24
Season
No. of Samples
Water Temp. °C
Secchi Depth-m
NH.-N
NOj+NO.-N
Inorg-TI
Kjel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a Turner Units
Productivity mgC/m3/hr.

No. of Samples
Water Temp. °C
Secchi Depth-m
NH--N
NO^+NO -N
Inorg_N
Kjel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P '
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a Turner Units
Productivity mgC/m3/hr.
Apr. -Nov.
8
24.8
0.9
29
139
168
356
495
6.9
14.4
21.0
35.4
15.1
8904
43
67
(29)
1
26.0
-
40
20
60
390
410
16.0
-
-
25.0
16.4

-
-
Dec. -March
9
9.3
0.7
65
228
293
224
453
17.6
30.4
38.4
68.9
8.4
1872
33
78
Gaston
3
10.8
0.9
75
280
355
217
497
8.3
22.0
11.5
20.0
18.4
5590
29(2)
-
John H. Kerr
2t 8, 14 John.H. Kerr 118, 1308
Apr. -Nov. Dec. -March Apr. -Nov.
8
22.8
1.9
61
66
127
224
290
7.2
16,2
3.4
19.6
14.6
2936
23
31
(30) Roanoke.
None
-
-
-
-
-
-
-
-
-
-
-
_
-
-
-
11
10.0
1.0
62.3
236
299
237
473
9.0
23.1
15.4
38.4
13.4
2508
27(6)
28(9)
Rapids
3
10,7
1.0
38
265
303
190
455
5.0
19.0
15.0
34,0
13.4
4109
35
-
5
26.3
0,7
54
88
142
650
738
24.0
36.8
71,6
108
7,7
29653
100
109
(44) Chowan.
16
25.8
0.7
63
137
200
677
814
15.0
35.6
44.4
80.0
11.4
3969
60
-
Dec. -March
6
10.5
0.6
245
134
380
1006
1140
51.0
80.0
103
191
6.4
25209
138
178
13.17.C01.SiLL
None
-
-
-
-
-
-
-
-
-
-
-
-
-
—
—
John H, Kerr 114, 111
Apr. -Nov.
5
23.2
1.1
57
23
80
336
359
7.0
7.0
12.4
23.4
15.7
9520
41
47
(44) Chowan
9
21.4
1.1
49
81
130
465
546
8.3
20.0
22.2
42.2
13.9
1846
28
™
John H. Kerr 108, 103
Dec. -March Apr. -Nov.
3
9.8
0.8
65
183
248
443
626
6.7
11.7
26.7
38.3
16.2
11799
85
102
MSI, R045.
None
—
—
—
-
-
-
—
—
-
—
—
—
-
—
—
7
23.8
1.6
34
45
79
285
331
7.1
9.3
9.4
18.7
19.4
3426
29
33
AL37
















Dec. -March
6
9.8
1.2
52
194
241
288
482
6.7
13.0
13.7
26.7
21.8
3340
36
43

















                                                                                              (Continued)

-------
APPENDIX A.
             SEASONAL MEAN VALUES OF LIMNOLOGICAL PARAMETERS, LAKE SAMPLES
             USED IN ALGAL ASSAYS Continued
Lake
Season
No. of Samples
Water Temp. °C
Secchi Depth-m
NH,-N
NOj+NO.-N
Inorg-w
KJel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a Turner Units
Productivity mgC/m3/hr.

No. of Samples
Water Temp. °C
Secchi Depth-m
Nil -N
NO^+NO -N
Inorg-N
Kjel-N
Total-N
PO -P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor £i Turner Units
Productivity mgC/m3/hr.
(31) .Crystal
Dec. -March
1
13.1
0.6
400
500
900
550
1050
250
390
20
410
2.5
1018
56
-
(38) _Lvtehes
1
14.8
1.2
80
230
310
280
510
30
45
5
50
10.2
99
19
-
(32) T^vlpfi
Dec. -March
1
14.4
0.9
510
400
910
900
1300
235
240
75
315
4.1
1393
27
-
(39)McKensie
1
16.0
0.6
30
40
70
360
400
5
10
15
25
16.0
374
37
-
(33) Finches (-34) Hodeina (35)Jarl
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APPENDIX A.
             SEASONAL MEAN VALUES OF LIMNOLOGICAL PARAMETERS, LAKE SAMPLES
             USED IN ALGAL ASSAYS Continued
Lake
Season
No. of Samples
Water Temp. °C
Secchi Depth-m
NH -N
NOj+NO -N
Inorg-N
KJel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor £ Turner Units
Productivity mgC/m3/hr.

No. of Samples
Water Temp. °C
Secchi Depth-m
NH.-N
NO;J+NO -N
Inorg-N
Kjel-N
Total-N
PO.-P
T-Soluble P
T-Particulate P ,
Total P
TN/TP
Phyto. Cell Density no. /ml
Chlor a_ Turner Units
Productivity mgC/m3/hr.
(3) Mattamuskeet
Apr. -Nov.
1
26.1
0.61
20
_
-
350
-
5
7
36
43
-
16068
44
-
(5)
1
25.5
0.61
15
24
39
350
374
5
11
6
17
22
2335
27
-
Dec. -March
None
—
—
-
-
-
—
-
-
-
-
-
-
-
—
—
Salters
1
15.0
0.92
35
10
45
290
30
5
14
1
15
20.0
3152
37
-
(4) Phelps
Apr. -Nov. Dec. -March
1
25.3
2.13
10
87
97
260
347
5
5
17
22
15.7
1279
23
-
(6)
1
26.0
0.92
10
31
41
250
281
5
10
14
24
11.7
430
36
-
None
-
-
-
-
-
-
-
-
-
-
-
-
-
—

Singletary
1
15.1
1.22
50
40
90
240
280
5
13
2
15
18.6
231
13
-
(1) Black
Apr. -Nov.
1
25.3
0.305
35
513
548
700
1213
50
165
50
215
5.6
1365
71
-
Dec. -March
1
15.0
0.24
90
600
690
920
1520
195
205
35
240
6.3
7944
80
-
(7) Waccamaw
1
25,1
2.13
15
7
22
380
387
5
5
12
17
22.7
348
29
-
1
15.2
1.22
45
10
55
320
330
5
9
11
20
16.5
650
18
-
(2) J
Apr.-Nov
1
25.0
1.22
20
50
70
250
300
5
5
8
13
23.1
454
20
-
(8)
1
26.5
>2.74
55
13
68
150
163
7
30
-
13
12.5
777
13
-
6nes"'(jo)
Dec. -March
1
15,2
0.92
40
45
85
280
325
5
12
8
20
16,3
2363
27
-
White
1
15.1
>3,05
30
15
45
110
125
5
9
1
10
12.5
62
8
-

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                                    TECHNICAL REPORT DATA
                             (Please read Instructions on the reverse before completing)
  1. REPORT NO.
        EPA-600/3-76-064
                                                            3. RECIPIENT'S ACCESSION»NO.
  4. TITLE AND SUBTITLE

   Evaluation of  the  Algal Assay Procedure
                5. REPORT DATE
                     June  1976
                                                            6. PERFORMING ORGANIZATION CODE
 7. AUTHOR(S)

   Dr. Charles M. Weiss
                                                            8. PERFORMING ORGANIZATION REPORT NO
 9. PERFORMING ORGANIZATION NAME AND ADDRESS

   School of Public Health
   University of North  Carolina
   Chapel Hill, North Carolina  27514
                1O. PROGRAM ELEMENT NO.

                  1BA031
                11. CONTRACT/GRANT NO.

                  R800399
 12. SPONSORING AGENCY NAME AND ADDRESS
  U.S.  EPA, OR&D
  Corvallis,  Engironmental  Research Laboratory
  200 S.  W. 35th St.
  Corvallis,  Oregon  97330	
                                                            13. TYPE OF REPORT AND PERIOD COVERED
                14. SPONSORING AGENCY CODE
                     EPA-ORD
 15. SUPPLEMENTARY NOTES
 16. ABSTRACT
        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.

        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 ji  and  productivity.

        The  algal assay procedure  provided an indication of  limiting nutrient, phos-
  phorus, 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 limit ing.
17.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b.lDENTIFIERS/OPEN ENDED TERMS  C.  COCATI Field/Group
 Algal Assay,  nitrogen, phosphorus,
  limiting nutrient,  trophic state,
  impoundments,  chlorophyll a
 3. DISTRIBUTION STATEMENT

 Release Unlimited
  19. SECURITY CLASS (ThisReport)
      Unclassified
                                                                         21. NO. OF PAGES
68
                                              20. SECURITY CLASS (Thispage)
                                                   Unclassified
                                                                         22. PRICE
EPA Form 2220-1 (9-73)
58
                                                           U. S. GOVERNMENT POINTING OFFICE: 1976-697-506/100 BEGION 10

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