STUDIES REGARDING THE EFFECT OF
  THE RESERVE MINING  COMPANY
   DISCHARGE ON  LAKE SUPERIOR
          SUPPLEMENT May 18, 1973
         U.S. ENVIRONMENTAL PROTECTION AGENCY
         Office of Enforcement anil General Counsel
             Washington, D.C. 20460

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STUDIES REGARDING  THE  EFFECT OF
  THE RESERVE MINING  COMPANY
   DISCHARGE ON  LAKE SUPERIOR
        ENVIRONMENTAL PROTECTION AGENCT
        Library, Region V
        1 North Wacker Drive
        Chicago, Illinois 60506
       SUPPLEMENT May 18,1973
  U.S. ENVIRONMENTAL PROTECTION AGENCY
         Washington,D.C.  20460

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3KVIROKM&ITAL PrCTECTICN AGENCY

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                      SUPPLEMENT CONTENTS*
 E.  Bennette Henson et al.,  The Ecological Effects  of
    Taconite Tailings Disposal on the Benthic Popula-
    tions of Wes tern Lake Superior	   1160

'"Armand E. Lemke,  Characterization of the North Shore
    Surface Waters of Lake Superior	   1320

 Joseph Shapiro, The Effects of Taconite Tailings On
    the Phytoplankton of Lake Superior	   1378
 *Further scientific works will be added to  the supplement as
  appropriate.

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                 THE ECOLOGICAL EFFECTS

              OF TACONITE TAILING DISPOSAL

               ON THE BENTHIC POPULATIONS

               OF WESTERN LAKE SUPERIOR



                     Prepared by:

        E.  B.  Henson
          University of Vermont
          Burlington, Vt.  05401

        E.  C.  Keller
          West Virginia University
          Morgantown, W. Va.   26506

        A.  J.  McErlean
          Office of Technical Analysis
          Environmental Protection Agency
          Washington, D. C.   20460

        W.  P.  Alley
          Zoology Department
          California State Univ. at Los  Angeles
          Los  Angeles, Calif.  90032

        P.  E.  Etter
          Office of Technical Analysis
          Environmental Protection Agency
          Washington, D. C.   20460
May 18, 1973
                          1161

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           The Ecological Effects of Taconite Tailings Disposal
           on the Benthic Populations of Western Lake Superior
                              E, B. Henson
                              E. C. Keller
                              A. J. McErlean
                              W. P. Alley
                              P. E. Etter
                           Errata and Addendum


Page    Line

1165      9    Insert the word "generally" between "Pontoporeia" and "require".

1165     12    Should read "b.  Pontoporeia requires ...".

1165     17    Insert "and perhaps detritus" after "bottom sediments".

1165     19    Delete "Reproduction occurs in the winter." and substitute
               "Reproduction occurs intermittently throughout the year in the
               deeper waters of the lake."

1189      1    Substitute "Pontoporeia" for "this organism".

1211      6    Delete "Alley and Anderson, 1968".

1213    Table
          D    "Greater than" symbol needed before "105 m".

1221      6    Insert "pelagic region of the" after the word "the".

1245           In the last sentence, delete "1939" and substitute "1949".

1254           Page 1254 should be inserted between pages 1257 and 1258.

1310      4    Insert "for the comparison of two means" between "test" and "for"

               For all correlation matrix tables (tables 0, P, Q, BB, CC, DD)
               a "-" sign means negatively correlated.

1318           Add the following reference to the LITERATURE CITED:

               Smith, Stanford 11., 1972.  Factors of ecologic succession in
               oligotrophic fish communities of the Laurentian Great Lakes.
               Journal Fisheries Research Board of Canada, Vol. 29, No. 6,
               pp. 951-957.
                                    1163

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                                 AND CONCLUSIONS
    1.  During the Pleistocene glaciation, a unique assemblage of aquatic
fauna was introduced into certain North American Lakes.  This fauna in-
cludes Pontoporeia andlfysis.

    2.  This assemblage of animals has been able to survive until today as
relicts or ecological remnants, in a small number of lakes in the Uniteti
States, including Lake Superior.

    3.  These species are considered to be endangered species.

    4.  The biology of Pontoporeia is summarized as follows:
        a*  Pontoporeia require temperature of less than 12°C, and
        generally reproduce during the winter in waters of less than
        6°C.
        b*  Pontoporeia requies well-oxygenated water.
        c.  The araphipod avoids strong illumination, and light controls
        its limnetic behavior.
        3-  Pontoporeia is adversely affected by turbulence.
        e.  Pontoporeia feeds on the microbenthos, consisting of bacteria
        and protozoa on the bottom sediments.  Adult Pontoporeia do not
        feed.
        f.  Reproduction occurs in winter.  The adults die scon thereafter.
        g.  Pontoporeia select silty-sand sediments having the following
            characteristics.
                                   1165

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        (1)  less than 5% clay, 10-20% silt, and 60-70% sand



        (2)  with median particle size of 50  3.5-4.0



        (3)  sediments of soft consistency



    h.  Pontoporeia undergo diurnal migration,  and are good swimmers.



        Reproduction takes place during these limnetic excursions,





    5.  The Lake trophic ecosystem consists of two dominant energy



pathways:



        a.  Phytoplankton-^-Zooplankton—>%sis~»Chubbs & Smelt—XTrout



        b.  Micro-benthos —>Pontoporeia—»Chubbs & Smelt—*-Trout



    6.  The benthos/ and particularly Pontoporeia are very important com-



ponents of the ecosystem.  Changes in populations of Pontoporeia will have



effects throughout the ecosystem.



    7.  Reduction of Pontoporeia populations will tend to:



        a.  Intensify competition among predators of Pontoporeia



        b.  Increase predation on Mysis



        c.  Reduce growth and abundance of fish populations



        d.  Increase the pressure on alternative food sources such as



        fish eggs.



        e.  Alter the composition of the benthic population structure and



        shift dominance relationships.



    8.  Statistical analysis of benthic data reveal that:



        a.  Prior to the discharge of tailings into Lake Superior, benthic



        population structure and densities or organisms were the same at



        sites above and below the discharge point.
                                   1166

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        b.  Following tailing discharge into Lake Superior,  PDntoporeia



        populations were reduced southwest of the effluent.



        c.  Population structures southwest of the discharge point also



        changed.



    9.  The release and deposition of taconite tailing markedly effect



the benthic ecosystem of Lake Superior.
                                   1167

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             Acknowledgement








The authors acknowledge the patience and





assistance of Ms. Grace Brown and





Mrs. Joan Conway in typing and preparing





this report.
                   1168

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Introduction



     Data discussed in this report have been briefly surrmarizcxi



previously and provided to the defendant's lawyers in a written



form (Plaintiff's Document No. 638).  In addition, the raw data



and reports, upon which the present discussion is based, have been



entered into evidence on several occasions.  Nevertheless, since



these data can be easily confused and because a nutrber of documents,



dealing with a single collection, have been produced and subse-



quently revised, the specific data sources are listed in Table A.



     This table shows that samples for benthic organisms have been



collected in 1949, prior to Reserve's operation, in 1968, and that



a large series of samples were taken during 1969.  Apart from the



first two collections, all successive samples have been taken solely



by Reserve personnel.



     Table A also gives the collection method or gear used, the spe-



cific number of samples, the extent of area sampled and the depths



and locations sampled.  Locations of the listed collections are



shown in Figure A.  It is important to note that the depths sampled



varied initially from opportunity sampling (unplanned) to stratified



sampling (all or most samples at a cannon depth of 200') in the



latter collections.  The earlier collections (#1 and 2) covered
                                1169

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1170

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                               TABLE A (Continued)

                             List of Documents Cited
Doc. Reference
    Number
      la        Burrows, C. R., 1950.  Second Annual Progress hepor>.  of
                Fisheries Investigations Conducted in Connection with
                Taconite Beneficiation Activities at Beaver Bay, Minnesota.
                Minnesota Department of Conservation, Division of Game and
                Fish.  Investigative Report Mb. 99, January 1950.

      Ib        Skrypek, J. L., C. R. Burrows, K. Bishop and J. B. Movie,
                1968.  Bottom Fauna off the Minnesota North Shore of Lake
                Superior as Related to Deposition of Taconite Tailings and
                Fish Production.  State ot Minnesota, Department of Con-
                servation, Division of Fish and Game in cooperation with
                Minnesota Pollution Control Agency,  Spec. Publ. No.  57,
                October 10, 1968.

      2a        "Resurie of Bottom Fauna Studies", dated August 11, 1969,
                under name of D. W. Anderson.

      2b        "Appendix G, Preliminary Investigations of Bottom Fauna
                Along the Southwestern Region of the North Shore, by David
                W. Anderson, Reserve Mining Company, Silver Bay, Minnesota"
                (13 pages).

      3         "Reserve Mining Company Memorandum.  To:  K. M. Haley,
                From:  D. W. Anderson, Subject:  Bottom Fauna Studies -
                June 24 - July 3, 1969.  Dated:  August 11, 1970".  Signed
                by D. W. Anderson   (9 pages).

      4a        "Reserve Mining Company Memorandum.  To:  K. M. Haley, From:
                J. C. Gay, Subject:  Bottom Fauna Studies, Statistical View-
                point, Dated:  May 25, 1970", under name of J. C. Gay
                (10 pages).

      4b        "Reserve Mining Company Memorandun.  To:  K. M. Haley, From:
                J. C. Gay, Subject:  Bottom Fauna Study, Statistical
                Analysis - Fall; 1969, Dated:  January 20, 1970; Itevised,
                February 26, 1970; Revised, March 25, 1970", under signa-
                ture of J. C. Gay   (20 pages).

      4c        "Bottom Fauna Study, Reserve Mining Company - 1969, dated
                April 30, 1970, unsigned (18 pages).

      5         "Preliminary Study of the Relationship of Organic Matter
                and Sediment Texture to Bottom Fauna Organisms-unsigned,
                undated   (2 pages).
                                     1171

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               Fig. A.  Map of Lake Superior showing approximate
                        along-shore distances sampled during the
                        periods 1948-49, 1968, and 1369. Arrow
                        indicates approximate location of the
                        tailing discharge
French R
                                     1172

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smaller geographic  distances along the shore compared to later



collections.  The total alongshore distance covered r'or all sampling



periods is approximately 50 miles.  In general, sampling effort has



been unbalanced with respect to the distance above and below the plant



outfall.  More samples have been taken southwest of the plant than



to the northeast.  (See Figure A)



     Comparisons of the various collections and analysis of poten-



tial effect are limited by several factors.  Not all collections were



made at the same time, location, nor at the same depth.  Different



numbers cf samples were taken at irregular time intervals and various



collection methods were employed.



     Methods for collection of field samples and the processing of



these samples in the laboratory have been detailed previously and



will not be reported in great detail here.  In general, the methods



and techniques are similar enough for intercomparisons with certain



stipulations stated.  Such comparisons, for instance, as numeric



density must be qualified by the statements that the Ponar dredge is



slightly more efficient than the Petersen dredge.  Petersen dredge



collections, therefore, tend to underestimate the true density of



benthic organisms and therefore invalidate rigorous intercomparisons



of Ponar and Petersen data.  Nevertheless, such comparisons as the



relative density at different sites along the shore within a given



collection period are valid.  Also, regardless of gear efficiencies, the



ratiometric or percentage associations among the various groups col-



lected are considered independent of the particular sampling method.
                                1173

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Thus, although the actual nuirber of animals collected by each method



may vary, the relative numbers within the different gr->npc are con-



sidered independent of sampling bias.  Analysis of this type of data



therefore should be limited to the relative, rather than the absolute,



numbers in a given species category.  Except for graphic presentation



in the earlier report wherein data from different methods have been



pooled, all statistical comparisons have been performed with technique-



constant or time-constant data sets or by use of data transformations that



consider relative rather than absolute differences.





Review and discussion of previous analyses and conclusions concerning



benthic organisms



     Many questions have been raised concerning the actual and possible



effects of Reserve's discharge on the ecology of Lake Superior.  In



order to place this in perspective, it is necessary to establish a



foundation for the conclusions reached by Reserve consultants concerning



effects on benthic organisms.



     The first benthic survey indicated in Table A was performed prior



to Reserve's operation with the stated purpose of defining baseline



conditions in the immediate area of the discharge.  This report



 (Document Reference la) stated that benthic organisms were "... sparse



both as to numbers and kinds of organisms.", that seasonal variation



was  lacking, that total numbers varied with depth and that "...



 {maximum densities] occur somewhere between 100 to 200 feet."  Numerical





                                1174

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data for this report are treated in detail below but .it r.,-y be inserted



parenthetically here that sanples taken at this time tv,;r^ cjenerally



honogeneous both in respect to total numbers and numbers within taxon



groups.  Thus,  8  years prior to the operation of the taconite plant,



benthic ocrnmunities were approximately nonogeneously distributed




within the study site and no above plant versus below plant difference?



were evident with respect to total numbers or to numbers within taxon



groups.



     In 1968  (Table A, #2) benthic sampling was performed by a coopera-



tive effort that included four state agencies and Reserve Mining Co.



This study encompassed a larger geographic area and also obtained a



larger number of samples than the 1948 study.  The final report



(Document Reference Ib) of the study made direct comparisons with the



earlier work and established various effects coincident with the



tailing discharge.  The salient findings of this report were as



follows:



     1)  "Numbers of Pontoporeia ... were significantly lower below



than above the plant ..."



     2)  "Numbers of oligochaetes were significantly higher below



the plant at  ... 100 feet but were significantly lower at depths of



250, 325 and 400 ft."




     3)  "Numbers of fingernail clams  [sphaeriidae] were ... higher



below the plant at 175 feet but not elsewhere."
                                1175

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     4)  "Numbers of chironomids ...  were ...  higher below the plant



at two of the five depths ..."



     5)  "Volume of bottom fauna ...  was significantly higher {below



plant] at ... 100 ft. ... but significantly lower at 325 feet."



     6)  " There is also seme indication that the deposition of fine



taconite tailings favor oligochaetes and chironomids although this



is not general at all depths..."



     7)  "It is likely that the significant differences ... can be



attributed to physical conditions associated with the deposition of



the fine portion of the tailings."



     8)  "... oligochaetes and chironomids were also more abundant



below than above the plant site in the 1949 ... [sample] ...".





     Ihis report discusses other aspects of the survey and also ex-



trapolates these findings to the cortmercial fish catch.  Although



later analyses have modified some aspects of these findings, the



majority of our work has reinforced and amplified these initial con-



clusions.



     The remaining studies  (Table A) performed solely by Reserve have



substantially failed to modify the findings noted above.  In addition,



many confusing statements have been made concerning the remaining



studies performed by Reserve.  For instance, in discussing a benthic



survey performed in April and May of 1969 and reported at the enforce-



ment conference, Reserve summarized its benthic findings as follows



 (Document Reference  2b):






                                1176

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           "1.  Based on preliminary data; total bottom fa1 ma



                densities are not diminished in zones :.•* tailings



                deposition.



           "2.  Bottom fauna population densities are materially



                lower southwest of Reserve's taoonite plant, wh£_re



                no evidence of tailings deposition was found, than



                the population densities associated with the



                tailings type bottom found in the vicinity of the



                Reserve plant.



           "3.  Based on the data presently available, it is not



                realistic to draw any conclusions as to the bene-



                ficial or detrimental effects of taoonite tailings



                on the bottom fauna along the North Shore of Lake



                Superior.



           "4.  This preliminary investigation has focused attention



                on areas needing further research.  Specifically,



                the casual associations of organisms to bottom type



                and the seasonal fluctuations in kinds and numbers of



                bottom organisms."





      Apart from the fact that these statements are not internally con-



 sistent and that they contradict the earlier work performed by the



 Company and state agencies, the data presented, when analyzed for the




present report,(see below pg. 15 ), yield significant correlations between





                                 1177

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the amount of tailings present (as determined by diemicaJ  analysis)



and the presence or absence of various taxa.



     A subsequent collection (Document Reference 3)  performed



during June and July of 1969 concludes as follows:



          "1.  There is a significant reduction in the total



               numbers of bottom fauna organisms 4.5 miles



               southwest of the plant.  This reduction probably



               extends to 7 miles southwest although its size



               is considerably diminished at this distance.



          "2.  At sampling sites southwest of the plant there are



               significantly fewer Pontoporeia than at the sampling



               sites northeast of the plant.  All of the differences



               between sites in these two general locations which



               may be formed have the same sign and many are sig-



               nificant at the 5% level.  There is, however, no



               apparent relationship between the number of



               Pontoporeia and the distance southwest  (up to 27 miles)



               of Reserve's plant.  This feature indicates that some



               natural factor(s) in the lake itself has a greater



               influence on Pontoporeia than the presence of the



               plant site.  If the discharge of the plant were in-



               fluencing numbers of Pontoporeia southwest of the plant,



               presumably it would result in an increasing gradient



               in the number of Pontoporeia as the distance from the





                                1178

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           plant increases in a southwesterly di lect-un,



     "3.   There is a significant reduction in the nunfoer of



           oligochaeta 4.5 miles southwest of the plant.  Other-



           wise the counts seem similar to one another at the



           other sampling sites along the shore.



     "4.   There are probably fewer Sphaeriidae at all sites



           southwest of the plant than those northeast.  This



           conclusion results from the fact that all the dif-



           ferences in counts that can be formed between the



           sites in these two areas have the saire sign although



           none are significant at the 5% level.  Again, the



           Sphaeriidae counts southwest of the plant shows no



           relationship to the distance from the plant.



     "5.   The nunfoer of Oiironomidae are probably significantly



           increased at sites immediately southwest of the plant,



           This nunfoer seems to diminish as the distance from the



           plant is increased."





     These findings directly contradict those stated earlier and



also introduce preliminary statistical analyses performed by the



Company.  Additionally, log transformations and moving averages are



applied to collection data without justification for the purpose of



"... [reducing] ... the influenos of random variation or counts
                                1179

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along the lake shore ...".



     Neither of these practices completely eliminates the large dif-



ferences noted previously with reference to above and below plant



comparisons.  For instance, for Pontoporeia, significant differences



(p. < .01) between above and below stations occur whether or not the



arithmetic or log values are used or moving averages are used.



     A detailed criticism of Reserve's analysis is contained in the



statistical section of the report.  The present discussion is aimed



at summarizing the viewpoints held by the defendant through tijne and



examining these for consistency and soundness of scientific value.



It is difficult not to be skeptical concerning Reserve's conclusions



and practices  with respect to the effects of the discharge on benthic



populations.  The Company's efforts with respect to benthic collections



have been heralded as the most extensive collections ever performed



on Lake Superior and this statement may have sane validity.  Yet, the



Company seems loath to accept the results of its own studies as the



conclusions of most recent reports show.



     In the most comprehensive and sophisticated study yet performed



(#5 in Table A), the Company glosses over any direct comparison of



above vs. below data or examination of population structure changes



and generally concludes they may not have sampled enough to draw con-



clusions.  These conclusions are part of a series of revised opinions
                                1180

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based on the same data.  It is interesting to oorcpcir^ successive




drafts of the documents that presumably result in the opinions given



in the final document  (Document Reference 4a).  It is necessary also



to question the logic of asserting that benthic populations are too




variable to measure, that not enough sapples have been taken to



assess potential effects and then conclude that the discharge is




having no effect.  One is also confused by the following statement:




          "One thing, however, that is significant about tailings



           is that the population is relatively uniform within




           the tailings area.  This probably is due to the uni-



           form substrate produced by the tailings which would tend




           to make the Pontoporeia normally distributed.  Past




           studies have shown that Pontoporeia tend to be normally



           distributed providing, however, that all other factors




           affecting their populations are uniform over this period.



           This is probably the true effect of tailings on



           Pontoporeia.  A very local effect which does exist is



           that bottom fauna populations are zero at the discharge



           point.  This is due to the heavy deposition or erosion




           in the immediate vicinity of the discharge point.  This



           effect is very local as shown by a fast recovery to




           uniform levels of bottom fauna populations."
                                1181

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     One is further confused in that, after establishing the required



sampling level that would permit estimates of lakewide affect (about



400-1,000 additional samples are recommended), Reserve has failed to



follow through on its own recommendation.  The final collection made



in September and October of 1969 (Table A) and the last benthic effort



known to us, consisted of only 27 benthic samples.



     Since that time, Reserve's employees and their consultants have



staunchly maintained that there is no effect of tailings discharge on



benthic organisms apart from a "localized" effect.  The extent of this



localized effect, its geographic limits and the overall effects upon



the ecology of the lake have yet to be measured.



     It is against this backdrop of confusion, assertion followed by



denial, and revision that the following analyses have been initiated.



What follows is divided into two main parts:  a discussion of the



biology and ecology of the Superior benthic organisms with an evaluation



of noted and possible ecologic effects; and a statistical examination



of available data.
                                1182

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    UNIQUE QUALITIES OF THE GLACIOMARINE FAUNAL COMPONENTS
       IN THE GLACIATED OLIGOTROPHIC LAKE ECOSYSTEM
WITH EMPHASIS ON THE AMPHIPOD PONTOPOREIA AFFINIS LINDSTROM
I.   The Presence of Pontoporeia jn i;he Glaciated ...
     Lakes of North America

 A-   The glacial event:

          It should be considered an established fact thet

     several thousands of years ago a massive global glaciation

     invaded deeply into the North Aiuerican continent, and

     extensively into the northern portions of Europe and

     Arctic Asia.  This glaciation made significant modi-

     fications in the morphology of the northern hemisphere,

     and effects of this glaciation persist to the present day.

          This Pleistocene glaciation was a unique event in

     the history of the earth.  These glacial events occurred

     so recently that rebound from release from the ice masses

     is still measured today.  During the Pleistocene epoch,

     the earth still experienced winters and summers, and along

     the southern borders of the ice sheets, the climate was

     moderate.  The major events of this global glaciation

     have been well studied and documented, both in North

     America and in Europe.  (Hough, 1958; Flint, 1957; Daly,

     1963).
                            1183

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B.   Introduction of a proglacial biota into North America:



          Associated with the Pleistocene and the glacial



     events in North America, an association of aquatic plants



     and animals of great significance was introduced into the



     continent.  Along the margins of the later ice fronts,



     there existed at various times a continuous series of pro-



     glacial lakes and connecting waterways.  There was,



     therefore, a water continuity extending arpund the Arctic



     Ocean from the Baltic Sea, across Europe and Asia, along



     the northern coasts of Alaska, through the chain of



     Canadian lakes, through Lake Superior and the other Great



     Lakes, the Finger Lakes of New York, to Lake George,



     New York, and over to Lake Champlain in Vermont.  An



     association of aquatic animals that were inhabitants of



     the Baltic Sea and the Eurasian coast were able to move



     eastwardly with the changed oceanic conditions  (£•<•[• /



     shallower water and reduced salinity) along the Russian



     coast, across the Bering Sea, and into the heart of



     North America to New England.



          This unique assemblage of plants and animals became



     established in the new lakes and waters as the  glaciers



     melted away, and they are known as glacial marine relicts.
                             1184

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Some of the members of this group are listed in



Table B.



     It was an extremely unique circumstance x.hat



allowed these events to take place and to introduce



this biota to our deepest and best quality lakes.



First, there occurred a climatic change that introduced



continental glaciation, and this so changed hydrographic



conditions along the northern coasts of Europe and Asia as



to induce genetic and ecologic races of aquatic species



in the coastal zones, and then, with the freshening of



the arctic waters, these species were able to colonize



eastwardly into North America; and because of the par-



ticular topography of Central Canada, the glacial margins



provided water continuity across the continent for the



dispersal of these populations all the way to New England.



After the glaciers departed, this association of animals



was able to survive as relicts or ecological remnants in



a small percentage of the lakes formed by the glaciers.



The number of these habitats suitable for survival is rela-



tively small, and numbers less than 50 lakes in North



America.   (Bursa and Johnson, 1967; Henson, 1966;



Muller, 1964; Ricker, 1959; Segerstrale, 1937a).
                        1185

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Table B  List of the Glaciomarine Relict Species
          Found in Some of the Glaciated Lakes
                Of North America
PLANTS:

     Silicoflagellata:

     Flagellata:
                Distephenus speculum

                Gymnaster sp.
ANIMALS:
     Insects:  Diptera
                Chironomidae:
     Crustacea:
               Copepoda:
               Schizopoda:
                Heterotrissocladius
                subpilosus Kieffer
                Epischura lacustris
                Limnocalanus macrurus
                Senecella calanoides

                Mysis relicta
Amphipoda:
                               Pontoporeia affinis
      Pisces:  Cottidae:
                Myoxocephalis quadricornis
 A number of  species that  are glacial relicts  in

 Europe are not  found in North America  (Muller, 1964).

 The  fingernail  clam  (Sphaeriidae) Pisidium  conventus

 may  have   traversed   from North America  to Europe  by

 by the same  route  as the  listed species  (Henson,  1966)
                        1186

-------
C.  The post-glacial environment:



         The retreating glaciers left behind them ands of



    lakes that could have harbored these residue.1, spscies.



    Though these species originated from marine or brackish



    environments, they had become adapted to a fxesh water



    environment.



         As the glaciers retreated, a warmer climate advanced



    into the once glaciated regions along with the native biota



    that had been forced south.  The shallower basins were



    filled with outwash materials, and only the deeper basins



    that maintained cold, well oxygenated water could retain



    their stocks of these relict species.



         The number of remaining lakes in the United States



    that harbor this relict fauna is now extremely limited.



    Only 19 lakes in the United States, including the five



    Great Lakes, are known to contain Pontporeia (Table C)



    and only 22 lakes are known in the United States that



    harbor either Pontoporeia or Mysis, or both.



         It is clear that the natural habitats for these unique



    species are being reduced  (Henson, 1966) or eliminated.



    Pontoporeia affinis and Mysis relicta, and the other as-



    sociated relict species should be considered endangered



    species.
                           1187

-------
Table c  List of Lakes in the United States that
         are known to have Natural Populations
         of Pontoporeia affinis and Mysis relicta
Location
Great Lakes :




Washington:
Wisconsin:

New York:







Vermont :

Name of the Lake
Lake Superior
Lake Michigan
Lake Huron
Lake Erie
Lake Ontario
Lake Washington
Trout Lake
Green Lake
Cayuga Lake
Seneca Lake
Keuka Lake
Canandagui Lake
Owasco Lake
Skaneatales
George Lake
Fayetteville
Green Lake
Champlain Lake
Sunset Lake
Pontoporeia
X
X
X
X
X
X
0
X
X
X
X
X
X
X
X
0
X
X
St. Catherine Lake x

Michigan:

Dunmore Lake
Torch Lake
Mullett Lake
0
X
X
Mysis
X
X
X
X
X
0
X
X
X
X
X
X
X
X
X
X
X
0
X
X
X
X
                      1188

-------
          The abundance and omnipresence- of th.is organism in


     the oligotrophic environs clearly indicate its great im-


     portance in the overall ecology of such id'-.-s,  Since


     this amphipod seems to be sensitive to pollution and


     eutrophication, then its presence or absence in a given


     locality can be used as a gauge in the determination of


     the quality of the benthic environment.  However, if


     Pontoporeia affinis is to be used as an indicator of en-


     vironmental changes, then its natural history as well as


     environmental relationships must be understood so that


     deviations can be recognized.


II .   The Taxonomy and Distribution of Pontoporeia


 A .    Taxonomy :


          Pontoporeia affinis was described by Lindstroru in


     1855 from samples taken from Swedish lakes.  The only other


     species in this genus is P. femprata Kr oye r .


          There is a certain amount of variation in the species ,


     and because the adult male doesn't fully develop until


     the last molt, it had actually been described as a separate

                       9
     species.  Segerstrale  (1971) after several years of de-


     tailed study, reported that these variants should not be


     considered as subspecies, even though we might retain the


     "subspecific" name for distinction.  Therefore, P_. hoyi,


                   , P. kendalli , and £. weltneri are all P_. affinis .
     The term P. affinis brevicornis refers to a smaller reduced
                            1189

-------
form of the species characteristic of the North



American populations.  (Bousefield, 1958; Henson,



1954; Larkin, 1948; Segerstrlle, 1937a, 1971a).
                        1190

-------
B.   Distribution:



         Pontoporeia affinis has a northern cit'-viv.v.jl^r



    distribution as follows:  Eurasian  The Baltic Sea,/



    brackish coastal waters of the Arccic coasts or Russia



    and Siberia, and as a glacial relict in the Baltic lakes



    in the glaciated regions; the lakes along the coasts of



    the Kara Sea, and down into the lakes oi Kamtchatka;



    and in the estuaries and lower reaches of the rivers



    draining north into the Arctic Ocean, and extending



    about 1,000 km up the Yenissei River in Siberia.



         North America:  Along the brackish coastal regions



    of Arctic Alaska, Ungava Bay, Canada, Hudson and James



    Bays.  Also in the St. Lawrence Estuary (Bousefield, 1955),



    It is found as a glacial relict in the colder oligotrophic



    lakes of central Canada, the Great Lakes, the Finger Lakes



    of New York, and Lake Champlain in Vermont.  The record



    by Norton  (1904) for Chamberlin Lake in Maine has been



    found to be an error.  The record for Lake Washington on



    the west coast has recently been explained on geological



    evidence (Segerstrale, 1971a).



         According to Ricker (1959) and Bousefield  (1958) ,



    Pontoporeia is found in the glaciated regions of North



    America  west of the Ottawa River but not north of the
                            1191

-------
St. Lawrence River, but recently, DadsweU  (parsonnel



coimnunication) has found it in a few lakes  in western



Quebec.  The known lakes in the United States that



contain Pontoporeia are listed in Table C.
                       1192

-------
III.   Biology of Pontoporeia affinis



           Although Pontoporeia are known to be err.: of the



      dominant benthic forms in the Great. Lakes, the available



      quantitative information concerning this species and



      the associated fauna comes chiefly from studies performed



      on Lake Michigan.  While no attempt is being made to



      equate these studies directly to all the Great Lskes,



      including Lake Superior, these findings are instructive



      with respect to that water body.



  A,    Reproduction



           The size of the adult male and female ranges from



      6 to 9 mm in Lake Michigan (Alley, 1968).  When the



      mature male develops during the last molt, it undertakes ,



      pelagic excursions, using its long antennae and special



      antennal sensory organs to locate mature females.  The



      mature male has an atrophied mouth; therefore, it does not



      feed, and dies shortly after copulation.  The adult female



      carries the fertilized eggs within large marsupial plates



      beneath her abdomen.  By the time the embroyos are re-



      leased, the female has degenerated almost to the point of



      being a suspended brood pouch, and dies shortly after the



      release of the young.  The young are slightly less than



      2 mm long when they are released from the brood pouch.
                             1193

-------
         Size frequency histograms,  collected throughout



    the 1964, 1965, and 1966 sampling seasons lor Lake



    Michigan, indicated that Pontoporeia living at a depth



    of 10 m mature in one year, those inhabiting a depth



    zone of 20-35m require two years to mature, and those



    living at depths greater than 35 m possibly require



    three years to mature.  At depths beyond 35 m, the



    amphipod population breeds intermittently throughout



    the year.  This basic pattern of growth and reproduction



    seems to be consistent with Lake Huron (Cooper, 1962),



    Great Slave Lake (Larkin, 1948)  and the Baltic Sea



    (Segerstrale, 1967).



B.   Feeding and nutrition



         The gut contents of several Pontoporeia collected



    from Lake Michigan and of several maintained in the



    laboratory were examined by Alley  (1968).  Aliquots of



    cultured algae were added weekly to the aquaria of the



    laboratory reared amphipods.  The  foregut of the labora-



    tory reared amphipods contained sediment coated with



    organic matter and intact algal cells.  While the organic



    matter on the sediment appeared to be digested, the cel-



    lulose walls and chloroplasts of the algal cells appeared



    to be little affected by the passage through the digestive



    system.  The foregut of the lake samples contained sediments
                            1194

-------
coated with a layer of organic material and an



occasional diatom frustule, while the. hinc.uUc, con-



tained only sediment and diatom frustules.



     Marzolf (1963) concluded from laboratory experi-



ments that the selection of a substrate by Ppntoporeia



was significant only when organic matter was present as



a surface film on the sediments.  He also found a



significant association between the density of this



amphipod and sedimentary bacterial numbers and the amount



of organic matter in the sediments.  Zobell and Feltham



(1942) showed that certain species of bacteria are capable



of degrading cellulose, chitin, and lignin which most



animals find difficult, if not impossible, to digest.



They earlier (Zobell and Feltham, 1938) established that



a variety of marine organisms can utilise bacteria as



an energy source.  Baier (1935) suggested that most



detritus feeders are nourished by the bacteria which de-



compose the detritus rather than by the detritus upon



which they appear to be feeding.  Marzolf (1963) found



that in the laboratory, Ppntoporeia actively selected



the sediments that had been "conditioned" by the growth



of bacteria.
                       1195

-------
         Pontoporeia are a very important link in the


    lake ecosystem.  They help to clean the bottom sediments


    by consuming the organic material and transferring the


    energy to higher trophic levels more available to the


    larger members of the community.


C.   Thermal relations


         Pontoporeia is generally considered to be an oligo-


    thermal species associated with cold water lakes? however,


    Alley (1968) found densities greater than 20,000 amphipods/

     2
    m  at 19° C. in Lake Michigan.  Henson  (1970) found similar


    water temperatures trends in the Straits of Mackinac.


    Samter and Weltner (1904) claim that Pontoporeia re-

                                                            o
    produces only at temperatures below 6° C.; and Segerstrale


    (1959) suggests an optimum temperature between 8-12° C.


         Segerstrale (1937) found 21-24° C. to be the upper


    survival limits on the basis of laboratory experiments.


    In some recent laboratory studies/ Smith  (1972) found


    12° to be the surviving temperature for a 12-hour exposure,


    but between 10-11° C. for a prolonged exposure.


D.   Oxygen requirements:


         Distributional records and field experience indicate


    that Pontoporeia requires an ample supply of oxygen to


    survive.  No definitive study has been  made of the oxygen
                             1196

-------
    levels in the lakes where the species is present, but


    generally they have about 50% saturation of oxygen


    Juday and Berge (1927), in an investigation of Green


    Lake, Wisconsin, found that PontopoJ:eia was able to


    survive within the sediments when the vater contained


    0.72 cc/1 dissolved oxygen one ureter above the bottom,


E.   Photic reaction


         This species avoids strong illumination.  Smith  (1972)


    cultured his controls exposed to only 1 ft. candle of

                           0
    illumination.  Segerstrale (1970, 1971) gives strong evi-


    dence that light has a controlling influence on the re-


    productive nature of the animal.  In the Finnish seas,


    the animal buries itself in the soft sediments in shallow


    water during the daytime, and becomes pelagic during the


    night.  Vertical migration is largely controlled by light


    (Marzolf, 1965b).


F.   Turbulence


         There is some evidence that Pontoporeia prefer still


    water.  Henson  (1970) observed that the species was


    not commonly present in high energy environments at any


    depth.  Smith  (1972) makes a special point of the adverse


    effect of turbulence on the animals.
                           1197

-------
G.   Substrate relationships:



         During the daylight hours, in the shallow, .inshore



    regions Pontoporeia remain burrowed in the surficial sedi-



    ments.  Marzolf (1965), in a laboratory study, found rhat



    Pontoporeia selected sediment particle sizes smaller than



    0.5 mm in diameter (sand) but did not discriminate between



    smaller sizes to a particle size of 0.0078 mm  (silt).



    Field observations from Lake Michigan indicate that



    amphipods are found in low numbers in clay sediments



    (average diameter of sediment particle size less than



    0.0039 mm) as well as gravel and glacial till  (Alley, 1968).



    This observation was verified by Henson (1970) when he



    found that the abundance of Pontoporeia approximated the



    normal distribution with respect to sediment type in the



    Straits of Mackinac, Lake Michigan.  Larger standing crops



    of amphipods were found in the silty-sand sediments, and



    in sediments containing 10-20 percent silt and 60-90 percent



    sand.  Substrates with a high clay content had relatively



    small standing crops of Pontoporeia.  This same trend was



    shown by Adams and Kregear  (1969) for eastern portions of



    Lake Superior.  They found that Pontoporeia dominate the



    sand bottom environment while oligochaetes were most



    abundant in the bedrock areas.
                           1198

-------
         These animals do not remain indefinitely in the



    sediments since some of the large juvenu.es, nature



    males, and mature females undertake excursions into



    the water column.  Wells (I960, 1963}  and Marzolf (1965)



    have reported vertical migrations for Lakz Michigan„



    Wells (1968)  showed that a small proportion of the



    Pontoporeia population are found above the sediments



    at depths greater than 36 m during the day.  Both



    Wells and Marzolf found that most amphipods remained



    in the sediments at night.  Size frequency distributions



    reported by Alley (1968) indicate that these vertical



    migrations seem to be localized and migrations do not



    extend laterally to any extent, at least in the nearshore



    environment where the water depths are 30 m or less.



H.   Organic carbon content of the sediments



         The percent organic carbon content of the Lake



    Michigan sediments shows that substrates which have the



    greatest concentration of Pontoporeia contain the lowest



    amount of organic carbon content (range of 0.06% to 0.71%),



    These areas of the lake which contain the greatest organic



    carbon content have few amphipods, probably because the



    organic materials were overlaying clay sediments which are



    not suitable for burrowing (range 3.02% to 4.02%).
                           1199

-------
Furthermore, the carbon compounds, found in the deeper


sediments, probably represent the terminal rain of


suspended particulate matter which is composed mainly


of undigestable cellulose, lignin, and chitin.


 Depth


     In the Lake Michigan study (Alley, 1968), 35


stations, located on five cross-lake transects in the


southern two-thirds of the lake, were sampled in tripli-


cate on a monthly basis from August to November 1964,


from April to November 1965 and from March to November


1966.  Stations of the B and D transect were not


sampled after June 1966 (Fig. B).


     A zone of maximum abundance of P. affinis occurred


between 30-60 m in Lake Michigan with the greatest


density of amphipods existing at the 30 m depth contour


(Fig. C). Powers and Alley  (1967) found the average


density at the 10 and 20 m intervals to be almost 4,000

           2
amphipods/m ; the 30 m contour to be greater than 8,500

           2
amphipods/m ; and the 50-60 m contours to be almost 6,000

           2
amphipods/m .  The counts at the 70 m depth contour dropped


to almost 4,000 amphipods/m  , and beyond 70 m, Pontoporeia

                                                      2
gradually declined in numbers to about 600 amphipods/m


at a depth of 270 m.  Mozley and Alley (1973) showed  that


a similar trend of Pontoporeia abundance occurs in


Lake Huron.
                        1200

-------
   CHICAGO
FIG. B   Index map of the Lake Michigan macrobenthos
stations of the long-term study area that were sampled
from 1964 to 1966.
                       1201

-------
I/) OJ
III  •\-
-j  2.
   I/)
   h-
   z
$
QJ
O
<
a:  Q
UJ  O
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9000-
8000-
 7000-
6OOO-
 5OOO-
4000-
3OOO-
 2000-
 1CXDO-
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      0  30  6O
                        90 120  150 180  210 240 270  300
                           DEPTH, METERS
     FIG. C .  Average numbers of amphipods/m pooled "by 10 -m depth
     increments vs . depth in meters .
                            1202

-------
     The interface of the sublittoral and profundai



environments seems to occur around the 35 n contour



in Lake Michigan.  This region corresponds roughi/



to the lower limit of the therF\ocline.



     Variations in Pontoporeia counts are considerably



greater within the sublittoral habitat because this



area is characterized by the greatest environmental



stresses and environmental heterogeneity.  Molar



action generated by storms and. bottom currents can



cause considerable shifting of the sublittoral sub-



strates.  In addition, bottom temperatures range from



1° to 19° C.



     Within the sublittoral regions of Lake Michigan,



for instance, Pontoporeia has developed two distinct



reproductive patterns that appear to be related to



the water temperature.  Amphipods mature in one year



in the warm, shallow, inshore regions of the sublittoral



and require two years to mature in the deeper portions



of this zone.  Pontoporeia of the sublittoral are geared



for a late winter-early spring period for reproduction.



     Pontoporeia reaches its maximum density at the



junction of the sublittoral and profundai zones and



this generalization probably applies to all the Great Lakes
                      1203

-------
The surficial sediments within this area are little



affected by turbulence, and the range of bottom tempera-



tures is not so extreme as in the shallower inshore



waters.  In this area and the adjacent profundal zone,



Pontoporeia matures and breeds intermittently throughout



the year.  There appears to be a greater proportion of



breeding females in the amphipod population that inhabits



this area and the contiguous profundal areas.



     The junction of the sublittoral and profundal zones



represents a reasonably stable environment where



Pontoporeia can effectively utilize the rain of suspended



particulate matter that is created through the enrichment



by upwelling, local runoff, and also the dominant along-



shore currents.  It must be emphasized that this junction



changes with seasons, and its location within an area



is dependent on the morphometric features of the lake



and the prevailing meteorological conditions.



     The profundal zone represents the most stable en-



vironment of the lake with all areas sharing a common low



water temperature.  The Pontoporeia of this region



probably require three or more years to mature, and



although breeding is intermittent throughout the year,



only a very  small portion of  the population  appears  to  be



mature at any particular time.  The density  of Pontoporeia



in this  area shows a strong inverse relationship with
                        1204

-------
depth, which probably reflects the lack or suitable



nutrients and the absence of an appropriate burrowing



substrate.



 Small scale patterns of spatial distribution and



 microassociations



     Short-term patterns of spatial distribution and



the microassociation of macrobenthic groups of Lake



Michigan were investigated about 2,QCO m off Mona Lake,



Michigan, at a depth of 18 m by Alley  (1968) and Alley



and Anderson (1968).  Systematic and random samples were



collected from a uniform fine beach sand by two divers



who sampled with a hand coring device.  The amphipod



population was composed of two juvenile size groups



of Pontoporeia (a 2 mm and a 7 mm group).  In this uni-



form sandy environment the amphipod population conforms



to the normal distribution (Fig. D).



     Interspecific microassociations, which were examined



by correlation analysis that compared the interaction



of large and small Pontoporeia with the major taxonomic



groups (Oligochaeta, Sphaeriidae, and Chironomidae)



showed that both size groups exhibited a significant



inverse relationship with Oligochaetes  (Figs. E, F, and G)



Large Pontoporeia also showed a significant inverse



relationship with the Sphaeriidae  (Fig. G) but neither



size group displayed any form of association with the



Chironomids.
                       1205

-------

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1207

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Number of ;3i)ha©riidae/Quadrat
FIG. G Association of juvenile Pontoporeia af finis ~7 nm in
length and Sphaeriidae of the short-term study area.
                       1209

-------
     It is often difficult to interpret the implications



of biological association because the term itself is a



loose definition which makes no distinction between re-



lationships derived from mutualism, parasitism, symbiosis,



competition, and predation with those based on similar or



dissimilar habitat requirements.  The patterns of assocja-



tion are also influenced by the size of the units from



which the samples are obtained.  The relationships between



organisms may vary from time to time and from place to



place.



     Segerstrale  (1965), in an investigation of the Baltic



Sea, found an inverse relationship between the successful



recruitment of the bivalve Macoma baltica and the abundance



of Pontoporeia affinis.  He also found a significant nega-



tive relationship between the abundance of the priapulid



worm Halicryptus  spinulosus and P. affinis.  Segerstrale



concluded that Pontoporeia ingested the spat of Macoma



and the eggs of Halicryptus as it fed on detrital material.



Although in Lake  Michigan the larvae of the Sphaeriidae



develop within the mantle cavity of the adult clam, the



immature individuals, when released from the brood  pouch,



are often small enough to be ingested by large amphipods.



Since  the feeding habits of Pontoporeia are not completely
                        1210

-------
    understood, it is difficult to determine if this



    amphipod actively seeks small clams or the c-ggs of



    the Oligochaetes when feeding in the substrates of



    Lake Michigan.



K.   Patterns of seasonal distribution



         In this detailed study (Alley, 1968; Alley and



    Anderson, 1968) , stations of the same transect were



    sampled on two successive days to determine if the



    standing crop of Pontoporeia varied from day to day



    at a particular locality or at a series of station.



    Statistical testing indicated that the standing crop



    remains reasonably uniform over short periods of time.



         Pontoporeia/ collected at the 35 stations, were com-



    bined into four depth zones:  10-35 m, 36-65 m, 66-105 m



    and greater than 105 m for the monthly sampling periods



    (Fig. K).  The 10-35 m zone was considered sublittoral.



    The 36-65 m zone represented the most nearshore area of



    the profundal and was greatly affected by terrestrial



    surface drainage and the nearshore aquatic environment.



    The 65-105 m zone represented a transitional area which



    was only partially influenced by the nearshore environ-



    ment, and the greater than 105 m zone was considered to



    be almost totally free of the nearshore influence.  The



    amphipod counts were converted to a square root trans-



    formation to facilitate statistical analysis.
                           1211

-------
             GEOMETRIC  MEAN NUMBER OF  POMTOPOREIA
CVI

                                                                                             in
                                                                                             
-------
     Pooled amphipod counts of the four depf^ zones

were treated statistically to determine if the standing

crops of amphipods were significantly different for the

three sampling years.  This test showed no apparent

yearly difference in the standing crop . values for the

10-35 m, 36-65 m, and the greater than 105 m depth zones.

It is difficult to interpret the significant yearly

fluctuations which occurred at the 66-105 m depth zone

because the differences do not seem to be the result of

a biological phenomenon nor does it represent a major

change in the aquatic environment.  The grand geometric

mean number of Pontoporeia for the four depth zones of

the three sampling seasons and the results of the

Kruskal-Wallis test are presented in Table D.
TABLE D  The grand geometric number of Pontoporeia a,ffinis
for the three sampling seasons.  H represents the results
of the Kruskal-Wallis test.	


Depth Zones	1964    1965    1966	H

  10-35 m                    62      55      67      2.78
  36-65 m                    78      75      75      0-52^
  66-105 m                   50      59      58      7.76
     105 m                   35      40      36      3.81
  All Stations Combined      53      55      54      0.55
   Significant at 5%
                        1213

-------
         A re-examination of Fig. H and Table D show that



    although there was  no significant seasonal variation



    in the abundance of amphipods  at the 10-35 m zone,



    there was considerable variation within comparable



    sampling periods.  In 1965 there was a general increase



    in the population from April to August/ followed by a



    slight decline from August to November.  The 1966 popu-



    lation decreased in numbers from March to April, in-



    creased from April  to August,  and then decreased sharply



    from August to November.  The numerical decrease from



    March to April 1966 represents the death of spent,  mature



    females.  The low density of amphipods in the late  spring



    and early summer is probably attributable to losses



    arising from the elutriation-screening device that  is



    used to separate the organisms from the sediment.  Large



    numbers of newly released amphipods may have been swept



    through the separating screen.  As the small amphipods



    increased in size,  they were less likely to be lost in



    this manner.  The remaining three depth zones do not appear



    to show any well defined patterns within the sampling periods.





L.   Patterns of standing crop



         All observations from each station of the five cross-



    lake transects were pooled for the three sampling periods



    to provide information of the distribution of standing
                           1214

-------
crop in certain shallow inshore stations  (Fig. I.} ,



located in the sublittoral :^one? such as A-l, A-6,



B-l, and B-8,is lower thai1, stations which are locate-.1



at. greater depths and farther from shore.  Furthermore,



the variation in counts rt these inshore stations is



considerably greater than at the offshore stations.



These inshore stations usually represent stations



of variable substrates or are close enough to the



shore to be affected by land-generated pollution.



Stations situated in the northern basin of the lake



have, in general, a greater standing crop than stations



located in the southern basin that are situated at



comparable depths.
                       1215

-------
                                                V   O
                                                (0
                                                (3
                                                O
 CO

 V
43
4->

 IH
 O
«*H

 co
• |H

.s
v<

•a

 rt
•1-1
 (U
 )H

 SJ
                                                    co
                                                    0)
                                                   ,4
                                                   4->


                                                    6
                                                    o
                                                to   u
                                                P!   fi
                                                w   rt
                                               •TJ  -(-"
                                               JJ   (0
                                               r4  4->
                                               H   °
                                                    4)
                                                    0)
                                               _:   rt
                                                    «j
                                                    M
                                                   4->

                                               o   «
1216

-------
                the Lake S uper igr Ecosyg t^rri
A.   Background on ecosystera dynamics



         Plants and animals living in a lake are intimate j.y



    inter-related with each ether and the environment.  No



    species is an isolationist.  What happens to one spectra



    will have some impact and effect on others; and if some-



    thing happens to a dominant species,, the impact on other



    species will be of even greater magnitude.  It has been



    shown in the earlier part of this discussion that



    Pontoporeia and the other faunal elements of glacioruarine



    relicts persist today as geologic remnants of former times.



    It is further emphasized that associations between and among



    these relict assemblages have also been fixed in time.



         The nutritional pathways form a significant portion



    of the ecosystem.  In simple terms, the ecosystem will



    consist of primary producers (plants, and the phytoplankton)



    that have the capacity to incorporate and transfer inorganic



    carbon (with the energy of sunlight, and nutrients in the



    water) , in the form of carbon dioxide or bicarbonates into



    organic carbons in the form of sugar or plant protoplasm.



    Food is made from non-food.
                            1217

-------
     These plants are eaten by animals at the next



trophic level (the herbivores/ zooplankton) f.hat transfer



the plant food and incorporate it as animal food.



Herbivores are then consumed by primary carnivores and



so on up the food chain.  This transfer of energy  (food)



up successive trophic levels is known as the trophic



dynamics of the ecosystem.  The nutritional, or trophic



inter-relationship is an extremely important facet of



the ecosystem.



     Certain events or human activities can greatly affect



the dynamic balance of an established ecosystem  (McErlean



& Kerby, 1972).  Consider a small food chain  (Fig. J)



consisting of 10 components (species).  Each horizontal line



represents a trophic level, and each higher level  obtains



its energy from the level below it.  The arrows indicate



the direction of energy flow and feeding specificity.



For example, in Fig. Ja (an illustrative model), species  (10)



feeds on both  (9) and  (8) while species  (7) feeds  exclusively



on  (4).  Assume that some stress is applied to  species  (4)



that might eliminate species  (4) as an adequate prey  for



species  (7).  If species  (4) is eliminated, this directly



affects species  (7) in that there is  no more  food  available,



and consequently, species  (7)  is eliminated.  But  also
                        1218

-------
 CO
 rn
 -n

 o
 TO
 rn


 CO
 m
 co
 en
TO


CO
—I
TO
m
CO
CO
          CO
          —I
          yo
          m
             ©'"
                                                                          (D
                                                                          1 I 7S

                                                                         71
                                                                        cn H-

                                                                            H\
                                                                        P- gi en
                                                                         rt
                                                                         (D O
                                                                         ft
                                                                         HI
                           1219

-------
    eliminated is  species  (6)  since it also has the restricted



    diet of species  (4).   Consequently, the structure of the



    ecosystem can  be reduced to the model shown in Fig.  Jb



    In this model  there  is less buffering capacity for now



    only three species support the system, and the stability



    of the system  is dependent on the availability and per-



    sistence of the  three  remaining species.  In Fig. Jb



    it can be seen that  all energy must now be passed through



    species (5)  to maintain the energy budget at previous levels



    The system now allows  fewer permutations and the out-



    come is dependent on the assumptions made.  For example,



    if species (10)  favored species (9), and species  (9)



    fed mostly on  species  (6)  and  (7), species (10) would



    therefore be forced to change its diet to species (8)



    which might be much more inefficient.  Therefore, species



    (10) in the long run might suffer/ and the population



    of  (10) would  be at a disadvantage.  This model will be



    applied to the situation in Lake Superior.



B.   The Lake Superior ecosystem



         The benthos of Lake Superior consists of Pontoporeia



    affinis, tubificid worms, the  fingernail clam Pisidium



    conventus, and the midge larvae of Heterotrissocladius



    subpilosus, and some lesser components.  However,



    Pontoporeia is by far the most abundant species  in  Superior
                           1220

-------
as well as in the other Great Lake (Eiltunen, 1969;



Henson, 1966; Powers and Alley, 1967; Anderson and



Smith, 1970)  .



     There are a large number of zooplankton species ia



Lake Superior that serve as herbivores in the lake,



but Mysis reljcta dominates in significance in the



ecosystem.  Among the fish species in the lake, the



major components are the chubbs (Leucichthys spp.),



the lake trout (Salvelinus namaYcush) , the deep-water



sculpin (Myoxocephalus quadricornis), and the burbot



(Lata lota maculosa).  Rawson  (1951)  has shown that



the commercially important benthic fishes:  longnose



sucker, Catostomus catostomus, and the white sucker,



Catostomus commersonii, feed directly on Pontoporeia.



Stomach analysis show that the diet of the longnose



sucker can be as high as 63 percent Pontoporeia while



the intake of the white sucker is about 30 percent



Pontoporeia.   Freshwater fishes of the family Cottidae



commonly known as "muddlers" in the Great Lakes, are



bottom dwellers that rely heavily on Pontoporeia as a



food source.   Some of these fishes represent important



food items of pelagic fishes such as the lake trout,



Salvelinus namaycush, and the American burbot, Lota lota,



(Hubbs and Lagler, 1959).






                       1221

-------
     Nevertheless, of the benthos, Pontoporeia is the



dominant species.  Pontoporeia feeds on the organic



detritus and the microbenthic community of the cottom



sediments, which includes large numbers of bacteria



and protozoa (Marzolf, 1965; Perrotte, 1971).  My sis



also feeds on the particulate organic matter and the



profundal zooplankton.



     Among profundal fish populations, the chubbs feed



almost exclusively on the Pontoporeia and Mysis  (Moffett,



1956; Wells & Beeton, 1963; Anderson and Smith, 1970).



Mysis and Pontoporeia comprise the major source of food



for the small (young) lake trout and the burbot as well



as the sculpin.  Pontoporeia and Mysis are therefore the



major source of  food for the young trout and burbot,



and the chubbs and sculpin.



     Many pelagic fishes, at sometime in their life history,



feed directly on swimming Pontoporeia.  Van Oosten et al.



(1938) have shown that juvenile lake trout and the American



burbot eat this  amphipod; Gordon  (1961) indicates that



the American smelt, Osmerus mordax, also eat Pontoporeia;



and finally the  diet of the common whitefish,  Coregonus



clupeaformig; which is one of  the most valuable  food  fishes



of the Great Lakes, consumes as much as 63 percent



Pontoporeia in its diet.
                        1222

-------
     The larger trout and burbot feed primarily on the



chubbs and this has a controlling effect o,: the chubbs.



The commercial fisheries constituted the next trophi-.;



level that harvested the trout, and to a lesser extent



the burbot.  [Added to this cast is the marine Ic-.irpzsy



(Petromyzon mar in us) that invaded Lake Superior sever.,-:.'



years ago.]



    A simplified scheme of the.-je i'ood relationships  is  illustra-



ted in Fig.jc. There are two main trophic pathways.  The  first,



derived from nutrient input, shows the energy pathway through



the phytoplankton, the zooplankton, Mysis, the chubbs,



and then to the lake trout and burbot, and then to man



and the lamprey.  The second pathway is derived from the



microbenthos, and leads through the Pontoporeia,  the



chubbs (and young trout and burbot, and the sculpin),



to the large trout and burbot, and then to man and the



lamprey.



     The introduction of the lamprey into Lake Superior be-



tween 1950 and 1960 has had an influence on the upper portion



of the system in that two predators now prey on the  trout.



In Lake Michigan the introduction of the lamprey  had a



major impact on the ecosystem.  The lamprey first eliminated



the trout, then shifted to the burbot, and after  both of



these fish were eliminated, the lamprey began attacking



the larger chubbs in descending order  (Moffett, 1956).
                        1223

-------
s

-------
In Lake Superior, the lamprey never .rained the same



degree of control of the system as it  ri I •<<> L^ke



Michigan.  The lamprey control program by the fedoraL



government appears to have been successful , but-, the



lamprey would continue to be a threat to the lake t.rc;'r



if present control measures fai.3 , or if the laruprey



begins to increase in numbers.  In a balanced system



where the lamprey does not significantly reduce the



trout population, the stress would not be transferred



down the energy pathways.



     On the other hand, stress exerted in the bottom



of the pathway system can propagate an effect through



to the higher levels.  It will be shown later that the



discharge of tailings into Lake Superior has had the



effect of reducing populations of Pontoporeia and



modifying benthic population structure.  Possible



effects on the ecosystem can be visualized as follows.



Again referring to Fig. Jc a reduction in abundance of



Pontoporeia will have two effects.  A reduction in the



standing crop of Pontoporeia, since Pontoporeia is the



major source of food for the sculpin, the chubs, small



trout and small burbot, as well as many other fish in



the lake (Anderson and Smith, 1970), may intensify the



competition for food among these several species of fish.




                       1225

-------
The more omnivorous fish will be favored, and there



would be more attention to Mysis as a source of food;



but Mysis may also be reduced in numbers because of the



turbid water.  In other words, there could be a localized



food shortage which would adversely affect the abundance



and growth of the chubs, trout, sculpin, and burbot.



There would be a selectivity of advantage and disadvantage



among the several species of chubs.  The intensified



competition for food could increase the incidence of



cannibalism among the fish, and cause an increase in the



consumption of fish eggs and other food sources.



     The reduction of Pontoporeia could also require ad-



justments in the feeding habits of the lamprey; the



lamprey, selecting the larger fish, would be likely to



shift to the larger chubs, as happened in Lake Michigan.



     Since Pontoporeia is the main grazer on the micro-



benthos, this niche will be open for other profundal



benthos, and the Oligochaetes would be the group to



increase in abundance.  Evidence presented in this report



shows that a decrease in Pontoporeia is accompanied by an



increase in Oligochaeta.  The worms are not the preferred



diet of the chubs, however, and an increase in the worms



would not be reflected by an  increase in the population



of chubs.
                        1226

-------
C.   Pontoporeia;  as an indicator of pollution and



     eutrophication



         Pontoporeia occupiers a unique position in the benr.hir



    environment of the Great Lakes,  Firstly, it is the moot



    abundant species in the sublit..tu
-------
adjacent to large metropolitan areas.  This disruption



of the benthic community was dramatically reported for



the western portion of Lake Erie by Carr and Hiltunen



(1965).  The dominant burrowing mayfly, Hexagenia spp.,



decreased to only one percent of its former abundance



while Oligochaetes/ Sphaeriids, and Chironomids signif-



icantly increased in numbers.  This trend in Lake Erie



seems to be sweeping further and further eastward as the



tempo of enrichment and pollution increases (Veal and



Osmond, 1968).  The deterioration of the benthic community



has now entered areas of the lake where the once-dominant



Pontoporeia are now nonexistent.  This characteristic pattern



of normal fauna displacement has been reported over wide



areas of the Great Lakes:  Cook and Powers  (1964) for the



St. Joseph River area, Lake Michigan; Ayers and Huang  (1967)



for the Milwaukee Harbor, Lake Michigan; Schneider et al.



(1969) for Saginaw Bay, Lake Huron; and Kinney  (1972) for



major nearshore areas of Lake Ontario.



     The exclusion of Pontoporeia from an area that it



previously occupied may be due to several factors operating



independently or in a combination.  If the  organic materials



found either within or upon  the substrate reach  a sufficient



concentration, then the aerobic microfauna  and microflora



can consume much or all of the dissolved oxygen  in the
                       1228

-------
nearbottom environment, directly suffocating more



sensitive species.  In addition, the r:rn,;r;-.ol properties



of the interstitial substrate environuent are .ui ,f.-.red



by anaerobic conditions, often resulting in the pro-



duction of toxic chemicals, such as hydrogen sulf.lde.



This newly created environment ^ s fairly stable and,



subsequently denies the sednnents to many burrowing



animals.  Organisms may also be eliminated from the



environment by the addition of poisons, such as bioci^es,



industrial wastes and domestic effluents.



     The abundance of Pontoporeia can also be affected



by changing the physical properties of the environment.



Alley (1968) showed that large juveniles in the sub-



littoral environment can tolerate large variations in



bottom temperatures  (a range of 1°-19°C).  However,



Sarnpter and Wiltner  (1904) observed that temperatures



greater than 7° C prevented egg production in Pontoporeia,



It is obvious from these findings that these amphipods



can be eliminated from any inshore area by a seemingly



slight elevation in water temperature simply because



they can no longer reproduce.  The disruption of the



substrate by accelerated sedimentation can also eliminate



or reduce the numbers of Pern.topo>reia, particularly if



the introduced sediments have the consistency of either
                        1229

-------
fine silts, clay, or have a grain size which is smaller



than clay.  These animals simply cannot burrow to any



extent into compacted fine sediments.  Their normal



feeding habits indicate that they ingest detritus



and sediments as they feed and this eating process



may possibly be disrupted by fine, compacted sediments.



Furthermore, if they cannot burrow into the substrate,



then they are openly exposed to their predators, and



are possibly consumed in greater numbers than would



normally be expected.



     The increased turbidity in the superficial waters



would be expected to inhibit the action of the phyto-



plankton which thereby reduce the productivity of the



lake and, in turn, reduces the contribution of organic



matter to the bottom sediments; and thereby, in a feed-



back, further inhibits the potential production of



Pontoporeia.



     The forerunners of the ecological disasters which



struck Lake Erie  (the decline of the important commercial



fisheries and the total collapse of the biological com-



munities of its western basin) represented minor changes



in the nearshore waters.  It was thought that these  changes



were insignificant and that pollution and eutrophication



were not  important in Lake Erie at that time.





                       1230

-------
     Introduction of the alewife into La>:«. Superjor



in about 1954 also deserves consideration as to the



possible role of this species in the ecosystem structure



of the lake.  The alewife is a problem fish and has



caused much concern and expense in some of the other



Great Lakes.  The main predator on the alewife is



the large lake trout and it has been well documented



that the lake trout plays a very significant role in



keeping the alewife population controlled (Smith, 1972) ,



Since young lake trout feed on Ppntoporei.a, any re-



duction in Pontoporeia will tend to reduce the lake



trout population, as was discussed earlier; and this,



in turn, may tend to allow alewife populations to



increase in Lake Superior.



     Pontoporeia occupies a critical and pivotal



position in the Lake Superior ecosystem.  No single



species in the lake, other than Mysis/ has such a



controlling influence on the entire biology of the



lake.  A reduction in the populations of Pontoporeia



and the other changes that have been documented in



this respect should serve as a warning that the lake



ecosystem may be in jeopardy.
                      1231

-------
Analyses of Lake Superior Benthic ftata



     Following the acquisition of the available data from



Reserve Mining/ the data were examined to ascertain the most



appropriate method of data analyses to answer specific ques-



tions and hypotheses to be tested.  Certain of the data sets



were not amenable to standard (parametric) statistical evalu-



ation due either to the fact that the question to be asked



was specifically directed towards the relative distribution



of benthic organisms or that the data were only reported as



average values, thereby precluding standard analyses because



of a lack of replication.



     The specific general question "asked of the data" was



concerned with whether statistically significant differences



among benthic populations were present as a function of



proximity to the Reserve Mining outfall or tailing deposition.



In the evaluation of the data, certain assumptions were neces-



sary and these were generally centered around the nature of



the question and the subsequent hypothesis being tested.



For the evaluations concerning the chi-square  (x2) tests,



the distributional problems of concern in benthic analysis



were not required, only reasonably moderate numbers of counts



or average counts in comparable categories.



     In the evaluation of the data by the method of linear



correlation, the basic assumption of random acquisition of  the



data and essentially a normal distribution  (bell shape) of



the two variables to be compared was of prime importance.






                          1232

-------
Certain non-linear relationships were evaluated by transfor-



mation of the raw data specifically by convr-r^ion to



logarithms or squares.  These transformations change non-linear



associations into linear  models so that they can be analyzed



by conventional parametric techniques.  For example, the



doubling phenomenon of many life growth processes is not



of a linear nature, witness the world human population gi-owth



curve  (over time).  If one wanted to examine this phenomenon



in a linear fashion, the log transformation could be used



to change a multiplicative relationship into an additive



relationship; to make it more easily understood and easier



to treat statistically.



     An analysis of variance (Snedecor, 1956) is a test devised



to evaluate differences which possibly occur among a series



of average values.  The underlying assumptions of the analysis



of variance (ANOV)are the relative normality (bell shape) of



the shape of the distribution of the variable being measured



(such  as the number of Pontoporeja).  The distribution must



be relatively normal in shape but this is not a rigorous re-



striction (Nissen and Ottestad, 1943) for the analysis of



variance.  The assumption of the randomness of obtaining the



sample is also necessary and it is our understanding that the



Reserve Mining data were gathered in this manner.
                          1233

-------
     The sampling designs used by Reserve Mining in these



studies were presumably structured to examine changes in



the benthic communities in the area of the Reserve outfall.



The hypothesis to be tested under these sampling designs was



based on the null hypothesis which states that among the



means no differences occur in the area under examination.



In other words, there should be no statistical differences



among any of the counts or measurements from any of the



sampled sites/ except for sampling errors, counting errors,



etc., and error due to natural variability which is charac-



teristic of living communities.  Further, the statistical



evaluations are made with a preselected value (critical limit)



that establishes the chances of accepting or rejecting the



null hypothesis.  The probability associated with this



critical limit of acceptance or rejection of the null hypothesis



is based on the number of samples and the number of means



being compared.  This probability is presented as P<.05 (<=



less than); P<.01 or P<.001 and denoted by:



           Probability Statement       "chances"^



          P<.05 but not <.01 = *     5 in 100



          P<.01 but not <.001 = **   1 in 100



          P<.001 + ***               1 in 1,000, etc.



We will also use the terminology  (N  S ) to represent no  sig-



nificant difference for probability  statements greater than



P =  .05 probability level.
                           1234

-------
General Methods of Analyses and Their General, Us_e

     The  X2  (Chi-square) test for contingency ^Goldstein 1964)


is used to evaluate the relative distribution of various


categories, such as, number of Pontoporeia, number of Oiigc-


chaetes, etc.

     Chi-square is computed by the following formula:
                       _
                       ~
where o = observed;  E = expected, and  £ represents the


summation of the categories being examined.  Table Ea,

presents hypothetical data to illustrate the use of


In this instance, geographic location is being examined


with respect to a fictitious DDT spill.  The calculated


value for the data given in Table  Ea  would be small and

presents no statistical reason to reject the null hypothesis;


that there is no difference between the two sets of data.


This would not be true if the collection data appeared as

in Table  Eb.  A calculated  x2  for these data would permit

an investigator to reject the null hypothesis and conclude

that the data are statistically different with a specified

level of probability.  Tables have been computed by theo-


retical statisticians to ascertain the critical limits of


acceptance or rejection of the null hypothesis:  the probability


associated with these critical limits is based on degrees of


freedom (DF) which may be generally explained as the number

of observations and the number of ways the data are arrayed.
                          1235

-------
Table  Ea
          Average Number of Animals Collected
Time                  In Eastern       In Western
Period              Portion of Lake  Portion of Lake  Total

Before DDT Spill          78               73          151

After DDT Spill           25               28           53

  Total                  103              101
Table  Eb
          Average Number of Animals Collected
Time                  In Eastern       In Western
Period              Portion of Lake  Portion of Lake  Total

Before DDT Spill          74               31          105

After DDT Spill           30               77          107

  Total                  104              108
                          1236

-------
Thus, each manipulation requires the loss of a degree of




freedom and for small data sets a larger-value '"or  x2



to be significant.  The acceptance of the nulx hypothesis



in chi-square analysis represents no significant deviations



between the observed distribution and the expected distri-



bution.  Rejection of the null hypothesis indicates that




something other than random chance has occurred and, in fact,



the observed distribution does not conform to the expected



distribution.



     Linear correlation analysis, which is computed by the



least squares method, tests for relationships among variables



two at a time.  The linear correlation coefficient  (r) can



range from -1 to +1 with 0 representing no relationship and



+1 a perfect positive relationship and -1 a perfect negative



relationship.  A positive relationship (+) indicates that



as one variable increases in magnitude so does the other



variable.  A negative relationship  (-) indicates that as



one variable increases in magnitude, the other decreases.



The significance of the correlation is based on the degrees



of freedom.  The null hypothesis to be tested for correlation



analysis will be that no relationships or associations exist



between the two variables being examined.  The hypothesis



being tested is again the null hypothesis:  that there is



no relationship, i.e., r=o.






                          1237

-------
     The analysis of variance (ANOV)  is used to examine



a series of means for the significance of a difference



(or no difference) between 2 or more means.  The null



hypothesis states that there is no difference between



or among the means.  The over-all ability distinguishing



differences between means is a function of sample size.



As sample sizes increase, one can detect relatively small



differences between means.  The critical level of acceptance



or rejection of the null hypothesis is based on two factors:



number of means being compared and the number of samples



used in the calculations of the means.  Tables have been



computed which provide values for these levels of significance.
                          1238

-------
Hypotheses Concerning Statistical and Ecological



     All hypotheses were directed towards determining whether or not




any significant modifications of the benthic environment occurred in



the area surrounding the Reserve outfall.  Data, which were obtained




from this area, were spread across a variety of transects.  The



selection of sampling sets was based on an experimental design



whose objectives were not clearly stated in the Reserve documentation.



It is difficult to ascertain the rationale of their benthic sampling



program from 1968 to date.  It is clear, however, that the basic




hypothesis under which they operated was, that no differences existed



within their sampling and natural fluctuations within this area



accounted for any differences seen.  The most recent argument of




Reserve Mining, that the data are unsound due to a significant dif-



ference between two types of sampling error, is unwarrented (unless



they specifically designed their sampling program to test the hypo-



thesis that variability among dredge samples was greater than vari-



ability within dredge samples).  An experimental design which focuses



on the variability among dredge samples is not the clearest way to



achieve the information desired about the effects of the taconite



discharge on benthic communities.  The only logical method to design



the ANOV tests is to use whichever sampling error is greater; in this



case, variation among different dredges.  This is the most conserva-




tive method of testing the hypothesis that no difference exists among






                               1239

-------
segments of the benthic cotmunities from site to site or above



outfall versus below the outfall.  In almost all analyses, our



evaluations have shown significant differences in abundance



(rejections of the null hypotheses) among the sites and above and



below the outfall.  And these differences have been detected using



the most conservative (the largest) estimate of sampling error



     Inferences about the numerical fluctuation of the taxonomic



groups as well as an evaluation of the quality of the benthic en-



vironment can be obtained from these statistical tests.  Statistical



methods generally stop with the acceptance or rejection of the null



hypothesis; however, the results of testing can be generally pro-



jected to some conclusion about the ecosystem under evaluation with



some degree of certainty.  This form of evaluation, which is coupled



with the experience  of aquatic biologists, is the analytical ap-



proach that we have used in arriving at the conclusions presented



below.



     Data partitioning  (i_.£-, allocation of effect to a supposed



cause or treatment) is a corrmon method of analysis which identifies



subareas of the data where the major differences occur.  Sometimes



internal partitions of a major data set are used, such as the Least



Significant Differences Test  (or the Tukey's Test) where an average



estimate of variation of sampling  is used as the basis for comparison



of major groups such as sampling sites or locations.
                               1240

-------
     Another method of data partitioning is to subdividr the data

into selected categories such as:  a comparison of u,-r,s above the

outfall with data below the outfall.    This partitioning can present

information relating to the status of specific geographical or

ecological regions.  The testing programs and conclusions presented

in subsequent sections will examine:  the inter-relationships which

exists between the benthic environment and certain physical parame-

ters such as distance from the outfall, percent tailings in the sub-

sediment and depth; differences in the abundance of taxonomic groups

above and below the outflow; and relative distributional differences

of the components of the benthic community.
         SUMMARY AND CONCLUSICNS OF SIGNIFICANT STATISTICAL
                 FINDINGS AND THEIR INTERPRETATION
Analyses of the 1949 and 1968 Data from the Survey of Benthos of
Lake Superior Near the Taconite Processing Plant (Document Reference
# la, lb).
     The general statistical methodology used in this evaluation was

the chi-square contingency test.  We are using this contingency test

to evaluate the relative distributions of taxonomic groups, i_.e_.,

Pontoporeia, Oligochaeta, Sphaeriidae, and Chironomidae (= Insecta)

across classification categories.  It is the purpose of these evalua-

tions to ascertain whether there were statistically detectable shifts

among the categories tested.  In evaluating these data with chi-square,

the 5% probability level was used as the minimum critical level.  Also,

the mean numbers of organisms were used as the data base since these
                               1241

-------
were the only available data.

     The results of the chi-sguare analysis show that no significant

differences were found between the combined average data obtained

in May, July, and September of 1949 for the two sites, one above and

the other below the outfall  (Table F and Figure  Ka  These results

indicate that the relative distributions of the organisms are sta-

tistically the same population when comparing the above vs. below

data of 1949.  This substantiates the results presented in the

Burrows 1949 and Skrypeck Reports.  (See Table A, Document Reference

# la, Ib)
           Table F
           Taxon
Mean Number of Organisms/m  by Taxon
Combined Data from 1949 (May, July,
September Samples)  Depths 55-434'


     1.7 miles above  2 miles below    Total
Bontpporeia
Oligochoeta
Sphaeriidae
Chironomidae
Other
521
282
36
55
9
502
308
38
72
9
1,023
590
74
127
18
           Total
          903
929
1,832
            *  for contingency  (calculated = 3.464)

            5% probability  X? critical value for four degrees of
            freedom = 9.49

            Hypothesis that the two do not differ  in their relative
            distributions is  accepted.  The distribution of organisms
            is probably the same.
                               1242

-------
     The data presented in Table G are for the same, gr-ari. >ral area



taken at a later date  (July, 1968) by Reserve Mining Ooirpany.



These stations were combined into above and below the plant data



sets and were examined statistically in the sapie manner as the




1949 data.  There was  a highly significant distributional difference



among the taxonomic groups surveyed in July of 1968 when one examined




the coirbined stations  above the plant vs. the combined stations below



the plant.
                              1243

-------
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                                                                   1244

-------
           Table G      Mean Number of Organisms/in^ by Taxon,
                        July 1968.  Depths Ranging Jrern j'-j' to 400'.
                        Combined data.
           Taxon            2 stations above    4 stations below    Total
Pontoporeia
Oligochaeta
Sphaeriidae
Chironomidae
Other
Total
761
379
112
24
2
1,278
194
550
69
53
1
867
-'55
929
18 1
77
3
2,145
           X2. for contingency  (calculated) = 322.727

           5% probability X2 critical, value for four degrees of freedom -
           9.49

           Hypothesis that the two locations are the same in their re-

           lative distributions is rejected.  The distribution of

           organisms is greatly different.


     Table H shows an evaluation of the total nuirbers of organisms from

combined data in 1949 vs. 1968.  The purpose of this test was to examine

whether there was a relative shift in the total numbers of organisms

above and below the outfall between the years 1949 and 1968.  The test

indicated there was a statistically significant shift in the relative

distribution of the total number of organisms above vs. below the

outfall between 1939 and 1968.
           Table.H      Mean Number of Organisms/m2 Combined Data
                        1949 vs. 1968.
                        Year       Above       Below       Total
1949
19fiR
903
1,278
929
867
1,832
2,145
                        Total      2,181       1,796       3,977
                                1245

-------
           X2 for contingency (calculated)  (with Yates correction)  =



           44.50



           5% probability X2  critical value for one degree of



           freedom = 3.84



           Hypothesis that the two stations do not differ in their



           relative distributions is rejected.  The distribution of



           organisms is greatly different.





     Tables I and  J are to be examined together.  The  X2 test results



to ascertain possible shifts in the relative distributions of the



organisms between the 1949 and 1968 for the same month (July) as sub-



divided upshore and downshore from the plant 1.7 miles and 2 miles,



respectively.  It is apparent that there is a shift in the relative



distributions of the organisms between the sites for both years.  The



two dates are different with regard to the relative distribution of



taxonomic groups, with the difference being ascribed, in part, to



differences in the frequency of Oligochaeta and Ghironomidae.  Chi-square



results are extremely significant in both cases well beyond the 5%



probability level.



     The magnitude of the July differences in 1968 is much greater



than those of July 1949; however, the important point concerns the



reason for the shift in 1949 vs. 1968.  In 1949 the shift was primarily



a function of considerably fewer Oligochaeta (proportionately) below the



outfall.  In 1968 the difference was due to an excess of Oligochaetes
                                 1246

-------
below the outfall which replaced the Pontoporeia \r\ dominance.



           Table  -     Mean Number of Organisms/m2 b\ T-xon, July 194S



           Taxon           1.7 miles above     2 miles below      Total
Pontoporeia
Oligochaeta
Sphaeriidae
Chironomidae
Other
Total
506
455
58
70
18
1,107
405
.164
43
45
4
661
911
619
101
115
22
1,768
           X2 for contingency  (calculated) = 55.52


           5% probability X  critical value for four degrees of


           freedom =9.49


           Hypothesis that the two Stations do not differ in their


           relative distributions is rejected.  The distribution of


           organisms is greatly different.


                                                  2
           Table  J     Mean Number of Organisms/m  by Taxon, July 1968



           Taxon           1.7 miles above     2 miles below      Total
Pontoporeia
Oligochaeta
Sphaeriidae
Chironomidae
Other
Total
613
415
109
23
1
1,161
254
535
105
73
1
968
867
950
214
96
2
2,129
            X for contingency  (calculated) = 137.76


           5% probability  X2critical value for four degrees of


           freedom =9.49


           Hypothesis that the two stations do not differ in their

           relative distributions is rejected.  The distribution of
           organisms is greatly different.
                                1247

-------
     The data of 1949 and 1968 for above stations (at approximately

1.7 miles) was oonpared (Table K) and showed a significant shift in

the distributions of the different groups.  Essentially, the data

showed that the Oligochaetes were lower in relative frequency in 1968

and the Pontoporeia and Sphaeriidae were greater in relative proportion

in 1968, oonpared to the distributions of 1949.
           Table  K     Mean Nurrber of Organisms by Taxon for July
                        data 1949 and 1968 for above (1.7 miles NE)
                        station.

           Tajon              1949           1968            Total
Pontoporeia
Oligochaeta
Sphaeriidae
Chironomidae
Other
506
455
58
70
18
613
415
109
23
1
1,119
878
167
93
19
           Total             1,107          1,161            2,268


           *  for contingency  (calculated) = 65.25

           5% probability -'X critical value for four degrees of

           freedom = 9.49

           Hypothesis that the two stations do not differ in their

           relative distributions is rejected.  The relative distributions

           of organisms are greatly different-between the two years.
                                1248

-------
     Table L presents the data for 1949 and 1968 from the sites

dcwncurrent  (SW) from the outfall.  It also cleat-ly 'le^r-strates

a significant shift between the distributions, notably the enormous

excess of Oligochaetes in the 1968 sample compared to the 1949

sample.
           Table  L     Mean Nutrfoer of Organisms by Taxon for July
                        Data 1949 and 1968 for below station (2 miles)
           Taxon              1949          J968           Total
Pontoporeia
Oligochaeta
Sphaeriidae
Insecta
Other
405
165
43
45
4
254
535
105
73
1
659
700
148
118
5
           Total               662           968           1,630
           X  for contingency  (calculated) = 214.79

           5% probability .X2 critical value for four degrees of
           freedom =9.49

           Hypothesis that the two stations do not differ in their
           relative distributions is rejected.  The distribution of
           organisms is probably different.
     In the examination of Figures Kb and L shows a clear indi-

cation of the major shift of the Pontogoreia population both in terms

of absolute numbers and relative abundance, as one approaches the out-

fall and proceeds away from the discharge point for data collected in

1969.  There is a corresponding shift in the Oligochaeta and
                                1249

-------
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-------
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                                  1.251

-------
Chirononidae populations.  This is dramatically reflected in the



shift in distribution of absolute numbers which starts downshore



from the plant.  This is also clearly indicated in the proportional



distribution of these t>ro groups of organisms.  A gradual increase



in the number of Sphaeriidae occurs after the 2.5 mile mark as one



proceeds southwest (downshore) from the plant.  There is an unstable



change in the Sphaeriidae and Pontoporeia as one proceeds downshore



from the plant with an increase of Pontoporeia at the final sampling



site 23 miles downstream.  These figures indicate an extreme imbalance



within the taxonomic groups which is the result of an environmental



stress.



     In Table M the mean numbers of organisms again are as examined



for data acquired in 1969.  The relative distributions are examined



now as a function of the miles from the plant, both upshore and down-



shore, so that both positive and negative values in terms of direction



from the plant are evaluated to see if there is heterogeneity or homo-



geneity of the relative counts along the 200 foot depth contour.  The



Chi-sguare test of the data in Table M clearly indicates an enormous



amount of heterogeneity in the benthic communities.



     When one examines the relative proportions presented in Table N,



one can see that the major community disruption occurs at 2.5 miles



southwest of the plant with Pontoporeia going from the relative pro-



portion of 68% to 15%.  Simultaneously, the Oligochaeta shift from
                                 1252

-------
8% to 41%, respectively.  Also of note in the shifts of the distribution



is the major increase in Chironomidae from northeast of the plant having



a relative proportion of 18% to a twofold increase (to about 42%) just



downshore fron the plant.  If some sampling sites had been placed



closer to the outfall on both sides, a clearer picture would undoubtedly



be presented in terms of the impact of taconite tailings on the general



ecosystem.  This same criticism applies to the nearshore sites and to



the problem of defining the limits of effect in the offshore direction.



Available data do not permit an estimate of the extent of effect in



this regard.  If Reserve's turbidity current functions as it has been



described, tailing effects on benthic populations must extend at least



to the limits of particulate deposition at a minimum, or to the 900 foot



contour.  It is naive, however, to assume that effects on the benthic



conntunity terminate discretely at the edge of the turbidity current



perimeter.  At the present time it is not possible to estimate the



entire area of effect due to the fact that sampling has not been per-



formed in these areas.  Table N, For clarity, presents the relative



distribution of the various benthic componentsT as percentages.  Ihe



major shifts occur near the outfall (between 2.5 miles NE and 2.5 miles



SW), and range from 68% to 15% for Pontoporeia and from 18% to 42% for



Chironomidae.
                                 1253

-------
scale were usually also correlated on a transformed scale.   In addition,



the significance of the correlation did not change with transformation;



variables that were inversely related on a linear scale were also in-



versely related on the transformed scale.  Analyses performed on trans-



formed data therefore generally reinforce and amplify the results



obtained in linear analyses.  Tables summarizing these results show



the significance of the correlations (+ or -); the notation "0" is



used to signify no significant correlation at the .05 level.





     Table 0 is a summary of all of the significant linear correlations



detected with at least P <  .05, from all of the June-July data.
                                 1254

-------
      Table  M     Mean number of organisms by Tajon/m2 at 200"  Depth
                   and Estimated Percentage of Tailings for
                   Various Stations Along Sampling Line 5.
                   Data for April-May 1969.
Miles fron
Plant	#Pont.  #01ig.  tSphaer.  #Chironi.  fTotal  %Tailings
9.0 NE
5.5 NE
2.5 NE
2.5 SW
5.0 SW
10.0 SW
17.0 SW
20.0 SW
23.0 SW
635
721
804
323
301
510
259
133
201
140
233
93
876
589
488
259
377
90
129
93
79
50
111
151
136
144
90
312
50
212
898
736
273
294
101
54
1,216
1,097
1,188
2,147
1,737
1,422
948
755
435
0
0
0
80
60
40
5
0
0
Total       3,887   3,145       983     2,930   10,945
       X2for contingency  (calculated) = 2850.25

      5% probability x2 critical value for 32 degrees of
      freedom =45.0

      Hypothesis that the two stations do not differ in their
      relative distributions is rejected.  The distribution of
      organisms is probably different.
      Table  N     Percentage of Organisms by Taxon/m2 at 200'
                   Depth and Estimated Percentage of Tailings.
                   Data Acquired April-May 1969.  These percen-
                   tage Data Refer to the Raw Scores Given in
                   Table  N.
Miles from
Plant
9.0 NE
5.5 NE
2.5 NE
2.5 SW
5 SW
10 SW
17 SW
20 SW
23 SW
Pont.
52.2
65.7
67.7
15.0
17.3
35.9
27.3
17.6
46.2
Olig.
11.5
21.2
7.S
40.3
33.9
34.3
27.3
49.9
20.7
Sphaer.
10.6
8.5
6.6
2.3
6.4
10.6
14.3
19.1
20.7
Chiron.
25.7
4.6
17.8
41.8
42.4
19.2
31.0
13.4
12.4
Total
100.
100.
99.9
99.9
100.
100.
99.9
100.
100.
%Tailings
0
0
0
80
60
45
5
0
0
                                                                    1255

-------
Correlations of Data for June and July 1969





     Data obtained from the Reserve Mining Company (Document Reference # 3)



for June-July were analyzed by the method of linear correlation with



corresponding scale transformations, to evaluate possible non-linear



effects.  The data are partitioned into the three data sets: upcurrent



(NE), dcwncurrent (SW), and all data combined.



     The abbreviations of the variables and their meaning are as



follows:



           SITE = 19 sites from NE to SW above and below the plant



           outfall.



           TOTOFG = the total number of organisms



           PCNT = number of Pontoporeia



           OSLIGO = number of Oligochaetes



           SPHAER = number of Sphaeriidae



           CHIRCN = number of Chironomidae



           PCTAIIS = the estimated percentage of tailings obtained



                     from estimates provided in the data report.



           VISTATTiS = the estimate of the presence or absence of



                      visible tailings as noted in the data report



           LOCMELE = the distance in miles above or below the outfall



                      (+ is above, - is below).



           TOTSQ = the number of total organisms squared
                                 1256

-------
           PCNTSQ = the nurrber of Pontoporeia squared



           QLIGOSQ = the nunber of Oligochaetes squared



           SPHAERSQ = the nunber of Sphaeriidae squared



           CHIEDSQ = the nunber of Chironomidae squared



           LTOTOR = the log of the total nurtber of organisms



           LPCNT = the log of the number of Pontoporeia



           LOLIQO = the log of the nunber of Oligochaetes



           LCHIHDN = the log of the number of Qiironomidae



           LSPHAER = the log of the nunber of Sphaeriidae



           REGION = sites northeast of the outfall vs. sites



                    southwest of the outfall (_i. e. above vs. below)





     No attempt will be made to present, in tabular form, all results



from correlation analyses performed for these data (Document Reference



3) and for data given in Document Reference 4.  The large number of



possible two-at-a-time combinations of the 20 variables examined on



linear, log and square scales and the possible permutations of linear



with other scales precludes the inclusion of these results.  Nevertheless,



the analyses have been performed and the results have been made available



to Reserve's lawyers.  For clarity and to conserve space only linear



combinations (PONT vs. PCTAILS, CHIRON vs. LOCMTIE, etc.) will be



treated in tabular form.



     With respect to the results of the non-linear combinations it should



be noted that variables that were significantly correlated on a linear
                                 1257

-------
Table  O
Correlation matrix for all linear variables analyzed
by regression analysis.  The following notations are
employed:  + = positively correlated,   = negatively
correlated and O = no significant correlation at
P   .05.  Document Reference 3,  all data combined.
—
s
1
T
E
—
0
P
c
T
A
1
L
S
—
0
+
V
1
s
T
A
1
L
S
—
+
0
0
L
O
C
M
1
L
E
+
0
0
0
O
T
O
T
O
R
G
+
—
—
0
0
0
p
O
N
T
O
0
0
0
0
O
0
O
0
L
1
G
O
+
0
0
0
0
0
+
0
s
p
H
A
E
R
0
0
+
-f
—
0
O
0
O
c
H
1
R
O
         R
         E
         G
          I
         O
         N
                                    1258

-------
The relationship of various components of the benthic community to



site is evident, all of the Pontoporeia estimates, both linear and



non-linear, were significantly negatively correlated with site as



one proceeds downcurrent past the effluent; there is a significant



decrease in numbers of Pontoporeia.



     Total organisms, whether measured on linear or non-linear scales,



were all correlated with position with relation to the effluent viz.



whether the samples are counted above the effluent or below the



effluent.  Since Region I was assigned to be above the effluent, these



analyses indicate that there are more total organisms above the



effluent than below.  (Table o)



     The numbers of Pontoporeia are strongly associated with region



and both linear and non-linear estimates of the Sphaeriidae.  One of



ifhe most important findings is the correlation of the number of



Pontoporeia with the percentage tailings:  significant decrease of the



numbers of Pontoporeia occurs coincidentally with the increase in the



percentage tailings.  (Table O)  The correlations of the numbers of



Sphaeriidae also showed strong associations with the region (above or



below the effluent) with more being present in the upcurrent sites



(Table 0).



     One of the strongest relationships detected in these analyses is



that of the numbers of Chironomidae with the percentage tailings and



the presence of visible tailings — as the percent tailings and visible
                                 1259

-------
tailings increases, the number of Chirononids also increases signifi-



cantly.  There is also a negative relationship between distance (SW)



from the effluent and the numbers of Chironomids (Table 0).



     Not surprisingly, the percentage tailings was highly correlated



with the visible tailings.  Percentage tailings was also highly



significantly correlated with region with a general absence of tailings



upcurrent and presence downcurrent (Table 0).



     The correlation analysis of the visible tailings also showed



significant positive correlations with the log of the nurrfoer of



Chironomids and a negative relationship with the region.



     Correlation of the location by mile again showed a strong relation-



ship with the log or squares of the number of Chironomidae.   Clearly,



this confirms the previous finding that as one moves into the downcurrent



effluent area, there is an increase in the number of Chirondmids.



     The correlation of the square of the number of Pontoporeia was



also highly significantly related positively to region and to the linear



and non-linear estimates of the Sphaeriidae.  PONTSQ is negatively



correlated with the site, IDCMILE and the percentage of tailings.



Again, this series of relationships show clear negative relationships



viz; as the percentage tailings increases, the number of Pontoporeia



decreases.



     The correlations of the squared transformation of the number of



Sphaeriidae are the same as the linear function of the number of
                                1260

-------
Sphaeriidae.  The correlations of the squared transformations of the



numbers of Chironomidae is similar to the linear function,  except



that there is an additional significant relationship with the log of



the number of Pontoporeia.  This could indicate that there  are signifi-



cant displacements of Pontoporeia by Chironomidae over the  range



encompassed by all of the data but not as a linear function of each



other.



     The correlations of all log transformations of all of  the count



data showed only two non-linear correlations (over and above those



associated on the linear scale) and these were:  the log of the number



of Oiironomidae with site  (negative) and the log of the number of



Pontoporeia with the square of the number of Chironomidae (negatively).



     Even though these analyses are clearly tests of relationships, it



is evident that the correlations indicate groupings of variables as a



function of above or below the effluent.  In short, the number of



Pontoporeia, the number of Sphaeriidae, and the total number of organisms



are positively related with the region under examination (higher above



and lower below).  Conversely, the visible tailings and percentage of



tailings are negatively correlated with location above or below the



outfall (lower above and higher below).





Correlations Below the Effluent




     Table P is a presentation of the linear correlation results of



downshore data only  (Document Reference # 3).  In this partition of the
                                 1261

-------
data, we examine the degree of relationship among the variables from



the zone of highest probable impact (at the effluent) to a "recovery"



zone some 30 miles downcurrent (SW).
                                1262

-------
Table  P
Correlation matrix for all linear variables analyzed
by regression analysis.  The following notations are
employed:  + = positively correlated,  = negatively
correlated and 0 = no significant correlation at
P   .05.  Document Reference 3, downshore  (SW) data
only.
         R
         E
         G
          I
         O
         N
O
s
I
T
I







0

P
c
T
A
1
L
s





0
—
+
V
1
s
T
1
L
S



0
0
—
—
L
0
C
M
L
E



0
0
O
0
0
T
O
T
O
R
G


0
0
0
0
0
0
P
O
N
T


0
0
O
0
0
0
0
O
L
1
G
O
O
0
O
0
0
0
0
0
s
P
H
A
E
R
O
—
+
+
—
O
0
0
0
c
H
1
R
0
                                   1263

-------
     The expectation should be of a gradual shift from the



effects of heavy deposition to an area of extensive recovery



for the macrobenthic organisms.



     The first correlations evident (those with site) show the



precise relationship in this graded zone.  The percent tailings,



presence of visible tailings, and all scales of the



Chironomidae are negatively correlated, i.e., as the site



numbers increase from a low of 7 at 2.5 miles SW to a high



of 19 at 30 miles SW, there is a decrease of the above variables



(Table P)• Hence, as one moves southwest from the outfall,



the percent tailings, visible tailings, and the number of



Chironomidae significantly decrease.



     The significant negative correlations of the numbers of



Chironomidae and site number, the log of the number of



Pontoporeia, and the distance from the effluent, along with



the high positive correlations of the number of Chironomidae



with the percent tailings and the presence of visible tailings,



is indicative of the magnitude and extent of the environmental



and biotic shifts of this graded impact zone.  The Chironomidae



significantly increased as the percent tailings and presence



of tailings increased? further, as one gets closer to the ef-



fluent  (from 30 miles SW), the numbers of Chironomidae increase.



Also, there appears to be a significant replacement of



Pontoporeia southwest of the effluent.
                             1265

-------
     The percent tailings significantly decreases as one pro-



ceeds further southwest from the effluent and all of the measures



of the number of Chironomidae (linear, log/ and square) increase



as the percent tailings increase.  The significant relationships



of the visible tailings are essentially the same as that of



the percent tailings.



     Clearly, the negative relationships of the mile location



downcurrent (SW) from the effluent show the most dramatic



association of the data set with the percent tailings, visible



tailings, and the series of Chironomidae counts, all decreasing



as one progresses away from effluent towards the southwest



(Table P) .



     Negative correlations of the log of the number of



Pontoporeia and the log and square transformations of the



numbers of Chironomidae were highly significant.  These non-



linear scales indicate the nature of the direct shift of the



decreased number of Pontoporeia to the increased number of



Chironomidae and that the number of Chironomidae are signifi-



cantly correlated to the percent tailings and the location down-



current  (SW) of the effluent.



     Table Q is a summary of the significant linear correlations



upcurrent  (NE) of the effluent wherein only the site and the



LOCmile  variables are correlated, negatively.  This is a meaning-




less correlation; however, it emphasizes the fact that the upshore
                             1266

-------
Table  Q
Correlation matrix for all linear variables analyzed
by regression analysis.  The following notations are
enplcved:  + = positively correlated,   = negatively
correlated and O = no significant correlation at
P   .05.  Document Reference 3, upshore data only.
O
s
1
T
E
0
0
.
P
c
T
A
1
L
S
O
0
0
V
1
s
T
A
1
L
S
0
—
0
0
L
0
C
M
1
L
E
0
0
0
O
0
T
O
T
O
R
G
0
0
0
0
0
0
P
O
N
T
O
0
0
0
0
0
0
0
O
L
1
G
^^
O
0
0
0
0
0
0
0
s
P
O
0
0
0
O
O
0
0
0
          R
          E
          G
          I
          O
          N
                                                           A
                                                           E
                                                           R
                                             C
                                             H
                                             I
                                             R
                                             O
                                   1267

-------
area is not subject to the same degree of environmental stress
as the area below (SW) of the discharge.
     These series of analyses indicate major changes of the
benthic communities which occur as a function of variables which
represent specific aspects of the physical environment associated
with some aspect of the Reserve Mining outfall; specifically,
mile distance from the outfall, visible tailings, percent
tailings, and site.  These analyses confirm the previous con-
clusion concerning the rejections of the null hypotheses from
independently gathered data that no differences existed among
the data gathered.  Hence, the extensive differences detected
above and below the outfall from previous data are validated
and the area of effect is shown to be even more extensive by
the present analyses.  These analyses clearly establish re-
lationships between components of the benthic community and
physical parameters which are direct functions of the Reserve
discharge.  These analyses also establish the shifts of the
communities among their components, especially Pontoporeia
being replaced by Chironomidae as the percent tailings increases
 (or as one becomes closer to the effluent).
     Recall that there exists an extensive series of relation-
ships in the data below the outfall and for the data as a whole
 (18 in the downshore  set and 40 in the  total data set) and
there is a lack of relationships in the upshore data  (one
relationship, only site number and mile location were correlated)
                              1268

-------
These factors are very strong indicators that in the more or
less "control" sample area upcurrent (NE), no significant
relationships exist since there is only random "natural"
variation present whereas for all of the data and the "recovery"
area (near SW) there are extensive modifications in both physical
and biotic factors which reflect strong relationships (that
are probably so subtle in the upshore data that they are not
detectable over and above "natural" variation) as a function
of the effluent and its dispersion.

Analyses of Summer/ 1969 Data Sets
     A series of analyses were completed on data obtained in
the summer of 1969  (Document Reference #3).  Replicated samples
were taken along four transects at the 100, 200, 300, 400, and
500 depth intervals.  The four transects (see Figs. M, N, 0, & P)
were located at 8.5 miles NE, at 2.5 miles SW, at 10 miles SW,
and one at 25.0 miles SW of the effluent point.  Only average
values were made available by Reserve Mining; hence, it was not
possible to complete rigorous analyses.  Correlation and
analyses were completed as well as graphic presentations for
clarity.
     Because of the limited data available and limited sites
examined, no generalized linear shore relationships will be
attempted.  These data were analyzed with the purpose of
                             1269

-------
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                                                    1270

-------
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                                         1271

-------
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-------
examining the depth profiles of the communities since the
linear distributions were already examined in the data pre-
viously discussed.  In regard to depth correlations, four
negative relationships were evident:  depth and the log of the
number of Sphaeriidae, depth and the log of the number of
Oligochaetes, depth and the log of the total number of organisms,
and depth and the log of the number of Pontoporeia.  The first
three were highly significant and the latter one, only moderately
so.  The number of these three types of organisms decline as
one sampled deeper and deeper depths, as did the log function
of their numbers.  Figures M, N, 0, & P  show the extent of
these declines as a function of depth.
     Although the relationships shown in these figures indicate
the general nature of the decline of the organisms, it is not
clearly indicative of the benthic community structure  (com-
position) .  Figure M gives a graphic representation of the
three-dimensional view of the relationship of the numbers of
Pontoporeia as a function of depth and distance from the ef-
fluent.  Unfortunately, the choice of sampling sites and the
absence of data at the outfall preclude an accurate specific
picture of the "true" nature of these interrelationships;
however, it is clear that if adequate data were available,
it would probably show a very marked influence of the effluent
                             1274

-------
on the depth and length profile of the numbers of Pontoporeia.
A "trough", located 2.5 miles SW of the plant, runs from the
200 through 500 feet zones.  This represents a considerable
shift in the generalized bathymetric features beyond the 100
foot depth zone.  Pontoporeia are not equally distributed across
the depths regardless of the mileage away from the effluent.
A visual intuitive test of the null hypothesis would undoubtedly
result in a rejection since clearly one can see that the
distribution is not homogeneous across either site (mile
location) or depth and appears highly modified near the effluent.
     Figure N is most dramatic in many respects showing an
enormous excess of Oligochaetes at the 400 foot depth at the
site closest to the effluent; of almost equal importance is
the excess number of Oligochaetes at the 100 foot depth, with
a large depression in numbers at the 200 and 300 feet depths
nearest the effluent.  Again, as with Pontoporeia  (Fig. M)
there is a very strong intuitive impression that these locations,
both in distance   (miles from the effluent) and in depth, are
obviously different above vs. below the effluent.
     Figure 0 shows that the numbers of Sphaeriidae are generally
similar to Oligochaetes except for the 2.5 miles/400 ft. depth
coordinate, which is the same sample on which the Oligochaetes
showed an excess.  Again, the data indicate a depression effect
near the effluent  (2.5 miles distant).
                              1275

-------
     In Figure P, the indication that the number of
Chironomidae increases as the percent tailings increases
(as predicted fro.u the earlier data analyses, Table P
is again evident at the 100, 200, 400, and 500 ft. depths.
The prediction is based on the previous 200 feet analysis
and because of the strong correlation with increase in
percent tailings as one approaches the outfall.  Again,
the visual impression clearly indicates that the numbers
of Chironomidae are not evenly distributed in the areas
sampled.
     The summary three dimensional figure  (Fig. G)
reflects the composite distribution of the total number
of benthic organisms reported.  Visual interpretation again
leads us to project that the average total number of benthic
organisms is not the same over the three-dimensional grid
examined.
                             1276

-------
              o
              o
              o
                             o
                             o
                             m
 O.
 
-------
Peaks and valleys are evident, with the mcst dramatic change
occurring nearest the outfall.  It is important to remember
that this nearest sample site is approximately 132,000 ft.
away from the outfall.  Closer sampling points obviously would
greatly clarify the modification effects of the outfall; how-
ever, if modifications of the order of magnitude shown in the
nearest site are occurring, by projection, one would expect
even greater community changes at the outfall.
     A visual overlay of these figures (Figs. M7 N, 0, and P)
indicates enormous disruption of the relative distributions
of these communities which strongly supports the highly signifi-
cant statistical results obtained earlier by the method of
Chi-square.
   Analyses of the Summer, 1969 Data (Document Reference #3)
     Several x2 tests were completed on the data from the depth
survey conducted in the summer of 1969 and illustrated in
Figs. M, N, 0, P, and Q.    The X2 for contingency test was
used to test whether the relative proportions of the numbers
 (counts or mean counts) are from the same statistical population,
i.e. , that the relative frequency of the counts among depths
above the plant are the same  as those below the plant.  This
series is divided into several separate tests in the major cate-
gories enumerated; specifically, number of total organisms,
number of Pontoporeia, number of Oligochaeta, number of Chironomidae,
and  number of Sphaeriidae.  Those organisms classified as "other"
were not analyzed due to their relatively small proportions.

                              1278

-------
                                    2
     In all cases, the calculated  x values were considerably

in excess of the critical tabular X2 value considered to be

attainable by chance alone (in this case, equal to 9.49, at

the 5 percent probability level).

     The following    tables (Tables R, s, T, U, V, W, X, Y, Z, & AA)

are self explanatory in regard to their specific tests.

The generalized conclusion is that  there are highly significant

inconsistencies among the distributions of the various benthic

components above vs. below the effluent.


          Table R      Chi-square for contingency evaluation
                       of the X total number of organisms
                       from samples taken 8.5 miles above
                       and 2.5 miles below the taconite
                       plant in the summer of 1969.  Compar-
                       isons involve the relative distri-
                       bution of the total number of
                       organisms for the five depths
                       (100, 200, 300, 400, and 500 feet).
                       Depth      Above      Below       Total
          F
          E
          E
          T
100
200
300
400
500
892
1,820
851
887
647
3,512
1,180
346
5,880
474
4,404
3,000
1,197
6,767
1,121
                       TOTAL      5,097      11,392      16,489

                       The calculated chi-square = 3764.442 with
                       4 degrees of freedom.  The tabular
                       critical x2 value at the 5% P level = 9.49,
                       The hypothesis that the two regions do not
                       differ in their relative distribution for
                       the five depths is rejected.
                            1279

-------
Table S      Chi-square for contingency evaluation
             of the X Pontoporeia from samples
             taken 8.5 miles above and 2.5 miles
             below the taconit'e plant in the
             summer of 1969.  Comparisons involve
             the relative distribution of the
             Pontoporeia for the five depths
             (100, 200, 300, 400, and 500 feet).

             Depth      Above      Below       Total

F            100          180        666         846
E            200        1,140        326       1,466
E            300          385         66         451
T            400          553         40         593
             -500          360      	98_         458

             TOTAL      2,618      1,196       3,814

             The calculated chi-square = 1185.024
             with 4 degrees of freedom.  The tabular
             "critical" X  value at the 5% P level =
             9.49.  The hypothesis that the two regions
             do not differ in their relative distri-
             bution for the five depths is rejected.
Table T      Chi-square for contingency evaluation
             of the X Oligochaeta from samples taken
             8.5 miles above and 2.5 miles below the
             taconite plant in the summer of 1969.
             Comparisons involve the relative dis-
             tribution of the Oligochaeta for the
             five depths (100, 200, 300, 400, and
             500 feet).

             Depth      Above      Below       Total

F            100          526      2,266       2,792
E            200          167        274         441
E            300          300         94         394
T            400          100      4,580       4,680
             500          141        129         270

             TOTAL      1,234      7,343       8,577

             The calculated chi-square = 2346.227 with
             4.degrees of freedom.  The tabular  "critical"
             X2 value at the 5% P level = 9.49.  The
             hypothesis that the two regions do not differ
             in their relative distribution for the five
             depths is rejected.


                   1280

-------
Table U      Chi-square for contingency evaluation
             of the X Insecta from samples taken
             8.5 miles above and 2.5 miles below
             the taconite plant in the summer of 1969.
             Comparisons involve the relative distri-
             bution of the Insecta for the five depths
             (100, 200, 300, 400, and 500 feet).

             Depth      Above      Below       Total

F            100          20         246         266
E            200         227         406         633
E            300         100         140         240
T            400         174         460         634
             500         100         206         306

             TOTAL       621       1,458       2,079

             The calculated chi-square = 93.160 with
             4 degrees of freedom.  The tabular
             "critical" X* value at the 5% P level =
             9.49.  The hypothesis that the two
             regions do not differ in their relative
             distribution for the five depths is rejected.
Table V      Chi-square for contingency evaluation of
             the X Sphaeriidae from samples taken 8.5
             miles above and 2.5 miles below the taconite
             plant in the summer of 1969.  Comparisons
             involve the relative distribution of the
             Sphaeriidae for the five depths  (100, 200,
             300, 400, and 500 feet).

             Depth      Above      Below       Total

F            100         166         334         500
E            200         286         174         460
E            300          66          46         112
T            400          60         800         860
             500          46       	4JL       	87_

             TOTAL       624       1,395       2,019

             The calculated chi-square =  503.290 with
             4 degrees of freedom.  The tabular
             "critical"  X2 value at the 5% P level =
             9.49.  The hypothesis that the two regions
             do not differ in their relative distri-
             bution for the five depths is rejected.
                  1281

-------
Table $      Chi-square for contingency evaluation
             of the X total number of organisms
             from samples taken 8.5 miles above
             and 10.0 miles below the taconite
             plant in the summer of 1969.  Com-
             parisons involve the relative distri-
             bution of the total number of organisms
             for the five depths (100, 200, 300,
             400, and 500 feet).

             Depth      Above      Below       Total

F            100          892      1,768       2,660
E            200        1,820      2,299       4,119
E            300          651      1,180       2,031
T            400          887        246       1,133
             500          647        133         780

             TOTAL      5,097      5,626      10,723

             The calculated chi-square - 1075.366 with
             4 degrees of freedom.  The tabular
             "critical" X2 value at the 5% P level =
             9.49.  The hypothesis that the two regions
             do not differ in their relative distri-
             bution for the five depths is rejected.
Table X      Chi-square for contingency evaluation of
             the X Pontoporeia from samples taken 8.5
             miles above and 10.0 miles below the
             taconite plant in the summer of 1969.
             Comparisons involve the relative distri-
             bution of the Pontoporeia for the  five
             depths  (100, 200, 300, 400, and 500 feet).

             Depth      Above      Below        Total

F            100          180        434         614
E            200        1,140        773        1,913
E            300          385        507         892
T            400          553         73         626
             500          360      	33_         393

             TOTAL      2,618      1,820        4,438

             The calculated chi-square = 711.829 with
             4  degrees of freedom.  The tabular
             "critical" x2 value at the 5% P level = 9.49,
             The hypothesis that the two regions do not
             differ  in their relative distribution
             fez the  five depths is rejected.
                   1282

-------
Table Y      Chi-square for contingency evaluation of the
             X Oligochaeta from samples taken 8.5 miles
             above and 10.0 miles below the taconite
             plant in the summer of 1969.  Comparisons
             involve the relative distribution of the
             Oligochaeta for the five depths  (100, 200,
             300, 400, and 500 feet).

             Depth      Above      Below       Total

F            100          526        828       1,354
E            200          167        933       1,100
E            300          300        180         480
T            400          100         53         153
             500          141      	40         181

             TOTAL      1,234      2,034       3,268

             The calculated chi-square = 537.971 with
             4 degrees of freedom.  The tabular
             "critical" X2 value at the 5% P  level =
             9.49.  The hypothesis that the two regions
             do not differ in their relative  distribution
             for the five depths is rejected.
Table 54      Chi-£quare for contingency evaluation of
             the X Insecta from samples taken 8.5 miles
             above and 10.0 miles below the taconite
             plant in the summer of 1969.  Comparisons
             involve the relative distribution of the
             Insecta for the five depths  (100, 200, 300,
             400, and 500 feet).

             Depth      Above      Below       Total

F            100          20         220         240
E            200         227         280         507
E            300         100         333         433
T            400         174         107         281
             500         100       	47_         147

             TOTAL       621       9,987       1,608

             The calculated chi-square =  262.989 with
             4 degrees of freedom.  The tabular  "critical"
             X2 value at the 5% P level = 9.49.  The
             hypothesis that the two regions do not
             differ in their relative distribution
             for the five depths is rejected.
                    1263

-------
Table AA     Chi-square for contingency evaluation
             of the X Sphaeriidae from samples
             taken 8.5 miles above and 10.0 miles
             below the taconite plant in the
             summer of 1969.  Comparisons involve
             the relative distribution of the
             Sphaeriidae for the five depths
             (100, 200, 300, 400, and 500 feet).

             Depth      Above      Below       Total

F            100         166        286          452
E            200         286        313          599
E            300          66        160          226
T            400          60         13           73
             500          46         13        	5£

             TOTAL       624        785        1,409

             The calculated chi-square = 103.850 with
             4 degrees of freedom.  The tabular
             "critical" x2  value at the 5% P level =
             9.49.  The hypothesis that the two regions
             do not differ in their relative distri-
             bution for the five depths is rejected.
                    1284

-------
     In an attempt to further clarify the changes of the benthic



community and to demonstrate the shifts in this part of the



ecosystem, data given previously were analyzed graphically.



Figure P, presents the relative distribution of the components



of the benthic communities at the 200 ft. depth and also for



all depths combined.  The two pie charts to the left of the line



represent those distributions above (upcurrent or NE) the ef-



fluent.  The exact distances for each site are given below the



pie diagram.  Pontoporeia represent the dominant benthic form



8.5 miles NE of the effluent "control or natural baseline".



If we compare this site with the site downcurrent from the



effluent  (2.5 miles/ SW, we see a dramatic shift within the



span of 11 miles.  Undoubtedly other changes are occurring of



a yery marked nature in the proximity of the effluent.



Note that the proportion of Pontoporeia never returns to



its upcurrent or "natural" level, even at 25 miles downcurrent



from the effluent.  Note also that, dependent upon the depth,



either Oligochaetes or Chironomidae "take over" or replace



Pontoporeia as the dominant taxa.  This is the clearest in-



dication of a dramatic change in community structure as a



function of distance downcurrent from the effluent.
                             1285

-------
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       O
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        o
                                                2
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M
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                                            a
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       V
       in
       6
                                                   I
                                                   a
                                                   y
                                                   J
                                                   «~»
                                                   i
                                                 y
                                                 z
1286

-------
            ANALYSES OF DATA FROM FALL 1969
             (Document Reference #4;a,b,c/


     The most extensive set of benthic data available from the

Reserve Mining files were obtained from a large-scale sampling

involving 5 sample sites/ with 5 or 6 replications being taken

at each site and with 5 subsamples being counted for each dredge

sample.  Sampling sites were located at 11 and 6 miles northeast

of the effluent and 2.5, 11, and 30 miles SW of the outfall.

Two types of analyses were performed on these data:  1) cor-

relation, and 2) analyses of variance.  As before, transformations

were performed on count data to ascertain the most appropriate

scale of evaluation of the results.  Again, the data were par-

titioned into three major subsets; data above (NE) the outfall,

data gathered from sites below (SW) the outfall, and all data.


Correlation Analyses Among all of the Data (Document Reference
#4;a,b,c)

     Table BB is a summary table of all of the significant linear

correlations found among the environmental and biological para-

meters.  As before, to conserve space, only the linear correlation

pairs are tabulated.  The general statements made concerning the

previous sets of data where regression analysis was employed apply

also to the present analysis.  Transformed data usually yielded the

same significant relationships and no changes in sign were noted.

These results are also available.  The position or location of
                            128?

-------
Table BB
Correlation matrix for all linear variables analyzed
by regression analysis.  The following notations are
employ?d:  + = positively correlated,  = negatively
correlated and 0 = no significant correlation at
P  .05.  Document Reference 4, all data conbined.
1 o — o
? + o
I cp Tl
T V
f I
ls I
1
L
S
—
0
—
0
L
0
c
M
1
L
E
+
—
0
—
—
T
O
T
0
R
G
+
—
—
O
0
0
p
o
N
T
O
O
O
+
o
—
o
0
o
L
1
G
O
0
—
+
0
—
0
0
+
s
p
H
A
E
R
—
0
+
0
—
0
—
+
+
c
H
1
        R
        E
        G
        I
        O
        N
                                                                R
                                                                O
                                    1288

-------
the sampling sites was positioned so that site 1 was located



most northeast of the effluent, and site 5 was the southwestern-



most site.  Except for percent tailings, all the significant



correlations with respect to site were negative.  These in-



cluded:  all of the measures of the number of Pontoporeia,



all of Sphaeriidae and the log of the number of Sphaeriidae



(Table BB).The three major indications of a significant shift



in the benthic community (number of Pontoporeia, number of



Sphaeriidae, and the total number of organisms), were all signifi-



cantly reduced as one progressed southwest from the point of



discharge.



     Table BB also shows the significant correlations with



region and the mile location of the sites. Three of these



correlations indicate essentially that the total number of



organisms decreases as one proceeds towards the southwest.



The number of Pontoporeia were negatively correlated with the



number of Chironomidae and with the percent tailings present.



These correlations suggest a major community shift from



Pontoporeia to Chironomidae (Table BB).



     The Oligochaeta, on the other hand, were also strongly



associated  (positively) with the Chironomidae, the Sphaeriidae



(positively), and were also positively correlated to the percent



tailings.  Hence, as the percent tailings increases, so do the



Oligochaeta, and as the numbers of Oligochaeta increase, so do



the numbers of Chironomidae and Sphaeriidae (Table BB).
                             1289

-------
     The Sphaeriidae were also negatively correlated with the
location of the site (positive), site number (negative), and
with the Oligochaeta and the Chironomidae.  The number of
Sphaeriidae increase as the percent tailings increases, and
they also increase along with the numbers of Chironomidae
and Oligochaeta (Table BB).
     Of all of the benthic component groups, however, the
Chironomidae are associated with a greater number of other
changes than any of the other benthic variables.  The number
of Chironomidae were negatively correlated with distance down-
current, the region, and all of the Pontoporeia variables
regardless of scale.  Positive correlations include the percent
tailings, and the numbers of Oligochaeta and Sphaeriidae.
Clearly, number of Chironomidae is the most extensively af-
fected variable in this table  (Table BB). Chironomids appear as
a replacement for Pontoporeia and are most highly associated
with the percent tailings, viz. where high percent tailings
occur, high numbers of Chironomidae also occur.
     From these and previous analyses, the major controlling
factor in upsetting the benthic community structure is the
percent tailings present.  All components of the benthic com-
munity are significantly affected by the percent tailings;
the numbers of Chironomidae, Oligochaeta, and Sphaeriidae,
all increase significantly as the percent tailings increases,
whereas the numbers of Pontoporeia decrease significantly as
the percent tailings increases.
                             1290

-------
     The correlations of the mile location of the station and
all components of the Chironomidae, Sphaeriidae, and Oligochaeta
were significantly negatively related, as is the number of
total organisms.  As one proceeds downcurrent from station one,
there is a significant decrease in the level of the numbers of
Chironomidae, Sphaeriidae, Oligochaeta, and total number of
organisms (Table BB),  The general explanation of this phenomenon
will be shown later to be explained by the extremely strong re-
lationship in only the three downcurrent  (SW) sites (Table CC).
     In regard to the variables and their non-linear transfor-
mations, all generally agree with their linear equivalents.
That is, tailings are correlated with numbers of Pontoporeia
in a simple linear fashion and with all the transformed variables
based on Pontoporeia, such as logs or squares.  This is probably
due to the fact that the data in this set (Table BB) were well
replicated and the entire raw data were available for analyses
which was not the case for the previous data sets where the
evaluations were made from the mean values.
     The significant correlations of region with most of the
benthic community components  [see Table BBJ numbers of
Pontoporeia (+), numbers of Chironomidae  (-), numbers of
Oligochaeta (-), and total numbers of organisms  (+)] indicate
again the magnitude of the differences.  These differences,
between the two regions  (up vs. downcurrent), will be more fully
examined by analyses of variance in a later section.
                            1291

-------
Correlation Analyses for Downshore Data (Document Reference
#4;a,b,c)'
     All downcurrent sites were organized into a single data
set for correlation analysis.  The general level and types of
significant correlations detected in these data sets strongly
reinforce the trends noted earlier as one proceeds downshore
(SW) from the Reserve Mining effluent (Table CC).
     The site numbers and LOCMILE (miles-distant from the
outfall) 3, 4, and 5 are strongly correlated with virtually
every linear and non-linear variable of the benthic community,
examined as well as percent tailings (Table CC).  These
associations are partitioned into two general groups:
1) those which increase with site number and LOCMILE,
and 2) those which decrease with site number or LOCMILE.
The strongest correlations obtained in these analyses with
site and LOCMILE were those with the percent tailings.
As one approaches the outfall, larger and larger percentages
of tailings are found.
                             1292

-------
Table  CC
Correlation matrix for all linear variables
analyzed by regression analysis.  The
following notations are eitployed:  + =
positively correlated,   = negatively cor-
related and O =*_ no significant correlation at
P_ .05.  Document Reference 4, downshore (SW)
data only.
          R
          E
          G
          I
          O
          N
0
—
1
T
E







0
~
P
c
T
A
1
L
s





0
0
0
V
1
s
T
1
L
S



O
0
—
0
L
O
C
M
L
E



0
—
+
O
—
T
O
T
O
R
G


0
+
—
0
4-
0
p
O
N
T


0
—
4-
0
—
0
0
O
L
1
G
O
O
—
+
0
—
0
O
•f
£«
p
H
A
E
R
0
—
-h
0
—
0
—
-!-
+
c
H
1
R
O
                                    1293

-------
The interrelationships of site number, LOCMILE, and average



percent tailings were plotted on the abscissa with the pro-



portion of Pontoporeia, Chironomidae, Sphaeriidae, and



Oligochaeta being plotted on the ordinates (Figs. S and T).



The reduction in the number of Pontoporeia is associated with



the replacement of this species by an increased number of



Chironomidae, with only a slight change in the number of



Sphaeriidae.  The distribution of the relative community



structures, shown graphically in Fig. U for these data, is



additional evidence of the strong nature of the interrelation-



ships which occur among the components of the benthic community



and the distances from the effluent and the percent tailings.



This segment of the analyses also demonstrates the major re-



lationships evident in previous discussions, except that the



strength  (nearness to a. perfect correlation) of the down-



current study is more persuasive than all previous studies.
                              1294

-------
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                             o -
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                       LU Z I—
                     1295

-------
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                 rt
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                                              3OVU3AV


                                              *      O
                                                                    LU
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                                                                               00
                                           uj Z
                                          1296

-------
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 CTi
^ en
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                                                                  n
                                                                  bt
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                                                                                                1297

-------
The benthic community structure shift for the 200 ft. contour
is verified by Fig. U. Pontoporeia comprise about 50% of the
community in the two sites above the outflow but less than that
at the two sites immediately downcurrent (SW).  Pontoporeia
becomes the dominant species 30 miles from the effluent location.
The data collected in the fall of 1969 show that massive dis-
tributional shifts in benthic population structure occur to at
least 11 miles downcurrent from the outfall.
     In short, all taxonomic groups are associated in one way
or another with the presence of the outfall and/or distance
from the outfall.  This clearly indicates the massive and
highly significant effect the outfall (percent tailings) has
on the structure and level of the benthic communities evaluated.

Correlation Analyses of the Fall 1969 Data for the Upcurrent
(NE) Sites Only  (Document Reference #4;a,b,'cT
     The summary given in Table DD of the significant correlations
found in the two upshore  (NE) sites reinforces the findings of
the previous correlation study  (Tables 0, P, & Q) that only a few
of  the upshore correlations were found to be significant.
The correlations of certain variables versus site indicate
that there might be a disruption of organisms at the near
effluent site, even though Reserve's data reports maintain
that no tailings are  found at the near effluent site.
                             1298

-------
Table  DD
Correlation matrix for all linear variables analyzed
by regression analysis.  The following notations are
enployed:  + = positively correlated,  =» negatively
correlated and 0 = no significant correlation at
P   .05.  Document Reference 4, upshore data only.
0
s
1
T
E
0
0
P
c
T
A
1
L
S
0
0
0
V
1
s
T
A
1
L
S
0
0
0
o
L
O
C
M
1
L
E
O
+
O
0
—
T
0
T
0
R
G
0
+
0
o
—
0
P
o
N
T
O
0
+
0
0
—
0
0
o
L
1
G
0
O
0
0
0
0
0
0
0
s
P
H
A
E
R
O
0
0
0
0
0
0
+
0
c
H
1
          R
          E
          G
          I
          O
          N
                                                                  R
                                                                  O
                                    1299

-------
These rather weak correlations are exhibited in Fig. V

where the changes in the relative values of these benthic

components are shown as a function of distance above the plant.

As one approaches the effluent from the northeast, there is

an increase in the total number of organisms present

(see Fig. V).  Likewise, there is an increase in the

number of Pontoporeia and Oligochaeta between mile 11 and mile 6

upcurrent (NE) from the effluent.  The relationship of the

location by mile to the community components is identical to

that of the site and is also evident in Fig. U.  The Oligochaeta

and the Chironomidae are also positively associated.

     To summarize the findings of these upshore analyses/ fewer

associations are noted and those that are  (with site and loc-

mile primarily) are a function of the general increase in the

total numbers of organisms (Fig. V),  The Chironomidae and the

Oligochaeta have a stronger association in these sites than

in the downcurrent stations that are under ecological stress.


Results of Analyses of Variance  (Fall 1969 Data - Reserve Mining
Report - Document Reference tt4;a,b,cT

     One of the most critical methods of examining data of the

type generated on these studies  is analyses of variance.  With

this test we can examine average differences among sites and be-

tween the two regions  (above and below the effluent).  As before,
                             1300

-------
 U. U.  —I Z> LU Z t-
                           \ <*'
                            • LLJ
                            * ^^>
                          O\ii

                           Vffi

                            0\'
                      Z
                      O
                      e*
                      X
                      u
              •t
H - 1 - 1 - 1
1 - 1
^—|—j.
                d3d
          3DVH3AV
                                  UJ
                                  U
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          1301

-------
we have partitioned the data into three groups;  1) data
collected below the outflow (SW), 2) data from above the outflow
(NE), and 3) all data.  The partitioning permits a clear inter-
pretation of modifications of the benthic environment.  In these
analyses, there are two kinds of errors estimated, as previously
discussed, dredge variation at the time of sampling, and
errors due to sample processing.  From practical experience,
we know that dredge sampling has a greater variation than
laboratory processing.  Therefore, we have chosen to use the
variation among dredge samples as the source of unexplained
variation.  This choice gives an extremely conservative statis-
tical test and it is proportionately more difficult to reject
the null hypothesis when using it.  Recall that the sampling
design was devised to test the null hypothesis, that no dif-
ferences exist among the sampling sites or regions  (above vs.
below).  Hence, the rejection of this hypothesis permits us
to  conclude that something other than random variation has
occurred in the environment being tested.
     Table EE is the summary table of the analysis of variance
from the Reserve data for the fall of 1969.  It is divided into
two parts,  the first tests for differences among the 5 sites,
and secondly, to tests for differences between  the  areas above
the outfall and below the outfall.  The columns marked "sampling
site" refer to the probability that the rejection  of the null
                            1302

-------
Table  EE
ANOV  Analysis for Variables at 200'  with
Probability < F Values for SITE and REGION
 (Document Reference # 4; a,b,c).
 All Data Combined
                    Sampling Site
                            Above vs.  Below
Variables
PONT
OLIGO
CHIRON
SHPAER
TDTORG
IPONT
LOIJEGO
LCHIRON
LSPHAER
LTOTOR
PONTSQ
OLIGOSQ
CHIROSQ
SPHAESQ
TOTOSQ
Prob.< F
SITE
0.0002
0.0001
0.0001
0.0500
0.0001
0.0002
0.0001
0.0001
0.0331
0.0001
0.0003
0.0001
0.0001
0.1002
0.0001
Prob. < F
REGION
***
***
***
NS
***
***
***
***
*
***
***
***
***
NS
***
0.0002
0.0223
0.0008
0.6280
0.0491
0.0004
0.5854
0.0055
0.8440
0.0273
0.0001
0.0129
0.0010
0.5657
0.0725
***
*
***
NS
*
***
NS
**
NS
*
***
*
**
NS
NS
                      1303

-------
hypothesis (no differences among the sites) is wrong.
For example (see Table EE)  for the number of Pontoporeia,
the "F" value has a probability of 0.0002 which means we
have only about 2 chances in 10,000 of being wrong in saying
that there are differences among the sites (over and above
sampling, laboratory analytical, and natural variability).
We, therefore, conclude that there are probably extensive
differences in numbers of Pontoporeia among the five sites
across the sampled intervals (Table EE).  Likewise, for
most of the linear variables, PONT, OLIGO, CHIRON, SPHAER,
and total organisms, there are significant differences among
the sites except for the number of Sphaeriidae.  When one
examines transformations to determine if there is some other
non-linear scale on which differences may occur, we generally
find the same set of statistical decisions as the linear series
with a few exceptions  (Table EE).  For the squared trans-
formations , four of the five variables examined showed high
probabilities of rejection of the null hypothesis.
                             1304

-------
     The salient feature of these linear and non-linear tests
is to examine the combined set of statistical decisions to
determine if patterns emerge.  The most consistent pattern
is simply that enormous differences are found in the composi-
tion of the benthic community regardless of the scale examined
(Table EE).  The number of Sphaeriidae do not change dras-
tically from one site to another.
     The picture that emerges in comparing above vs. below
effluent data can be generalized, but it is not the same as
the pattern which emerged in the site analyses (Table EE).
In the linear phase of the analyses/ there were the same
set of statistical decisions (not as strong in the case of
the number of Oligochaeta or total organisms) but nevertheless
these differences are distinct and unmistakable.  When we
examine the general pattern, the number of Pontoporeia and
number of Chironomidae are extremely different in the regional
comparisons.  Both the number of Oligochaetes and the total
number of organisms are significantly different in the linear
phase and the log of the total number of organisms is signifi-
cantly different, as is the square of the number of Oligochaeta,
                             1305

-------
The generalized pattern of differences falls into three
groups:  1) highly significant difference (number of
Pontoporeia and number of Chironomidae, 2) significant but
not strongly so (number of Oligochaeta and the total number
of organisms)/ and 3) not significant  (the number of
Sphaeriidae).   The general statement of conclusion is:
significant differences in numbers of Pontoporeia,
Chironomidae,  Oligochaeta, and total numbers of organisms
exist above and below the outfall.
     Table FF represents the analyses of variance for various
parameters of the three sites downcurrent (SW) from the ef-
fluent at the 200 ft. contour.  Since all of the null hypotheses
were rejected at high probabilities, it will suffice to state
that extensive differences exist for all variables among the
three sites downcurrent (SW) of the effluent.  The magnitude
of these differences clearly indicates that the sites are not
homogeneous and, indeed, some reasons other than natural,
random, or sampling variation must be  responsible for these
differences.
                             1306

-------
Table  FF    ANOV  Analysis for Variables at 200' with
             Probability < F Values for Differences
             Between Sites (Document Reference # 4;  a,b,c)
             All Downshore Data/ Fall 1969
Variables
PONT
OLIGO
CHIRON
SPHAER
TOTORG
LPONT
IDLIGO
LCHIRDN
ISPHAER
LTOTOR
PONTSQ
OLIGOSQ
CKEROSQ
SPHAESQ
TOTOSQ
Prob. < F
SITE
0.0001
0.0001
0.0001
0.0041
0.0001
0.0001
0.0001
0.0001
0.0043
0.0001
0.0001
0.0001
0.0003
0.0049
0.0001

***
***
***
**
***
***
***
***
**
***
***
***
***
**
***
                      1307

-------
     Table GG shows ths summary for analyses of variance
in the comparison of the two upshore sites for the Fall 1969
data.  The general pattern is one of similarity, since only
6 of the linear and non-linear variables are significant and
none of them is highly significant.  As pointed out previously
in the correlation analyses, there is a slight but signifi-
cant increase in the Oligochaetes and the total number of
organisms from 11 miles NE to 6 miles NE of the effluent.
The general pattern above the effluent is a significant
increase in the total number of organisms and the number of
Oligochaeta.
     The general summary of the analyses of variance points
to three general conclusions:  1) there are extensive dif-
ferences in the number of benthic organisms above vs. below
the effluent; specifically, the total numbers of organisms,
the numbers of Pontoporeia, the numbers of Oligochaeta, and
the numbers of Chironomidae; 2) differences exist among the
three downcurrent  (SW) sites for all of the benthic components;
and 3) minimal differences occur between the two sites above
the effluent point.
                             1308

-------
Table  GG    ANOV  Analysis for Variables at 200'  with
             Probability < F Values for Differences
             Between Sites (Document Reference # 4; a,b,c)
             All Upshore Data, Pall 1969
Variables
PONT
QLIGO
CHIRON
SPHAER
TOTORG
LPONT
IDLIGO
LCHIRCN
LSPHAER
LTOTOR
PONTSQ
OLIGOSQ
CHIROSQ
SPHAESQ
TOTOSQ
Prob.< F
SITE
0.1355
0.0124
0.7684
0.5298
0.0327
0.1607
0.0186
0.6199
0.6214
0.0351
0.1143
0.0143
0.7333
0.7062
0.0348

NS
*
NS
NS
*
NS
*
NS
NS
*
NS
*
NS
NS
*
                      1309

-------
              EVALUATIONS OF RESERVE REPORT #5
         FOR DIFFERENCES ABOVE vs. BELOW THE OUTFALL
     Evaluations were also completed for data reported in

Table A as Document Reference #5.  The evaluations were con-

ducted by  x2 for contingency for the overall data and dis-

tributional data and by the t test for the number of

Pontoporeia.  The t test showed that there was a highly

significant difference between the two means  (X above =

47.09 and the X below = 31.31).  The  x2 value for testing

the relative distributions of the components of the benthic

community above and below the effluent are presented in

Table HH.  Again, the results of this test show that there

is a major (significant) shift in the composition of the

communities above vs. below the effluent.

     In Table II is presented the set of data for the dis-

tribution of the data among the 5 sites.  As before, ex-

tensive differences exist among the relative distributions

of the benthic community for the two sites above vs. the

three sites below.
                             1310

-------
Table HH         Chi-square for Contingency to Evaluate
                  the Relative Distribution of Numbers of
                  Organisms Above and Below the Reserve
                  Outfall.  Data for September, October 1969.
                  Pont.	Oligo.	Chiron.   Sphaer.
.above
Below
Total
518
501
1,019
227
398
625
78
178
256
210
291
501
1,033
1,368

Chi-Square = 53.5, 3 degrees of freedom.  Highly significant

P < .05.  Reject null hypothesis.
                     1311

-------
Table II          Chi-Square for Contingency to Test the
                  Relative Distribution of Numbers of
                  Pontoporeia, Oligoohaeta, Chironomidae,
Sphaeriidae Above and Below the Reserve
Outfall. Data for September, October 19<
(Document Reference # 5) .

11 miles (NE)
6 miles (NE)
2.5 miles (SW)
11 miles (SW)
25 miles (SW)
Pont.
249
269
114
265
172
Oligo.
162
125
120
236
42
Chiron. Sphaer.
42 107
36 101
90 103
80 118
8 70
Chi-Square Value = 200.56  12 df. x2 highly significant.

Reject null hypothesis.
                      1312

-------
Summary of Statistical Analyses
     Statistical analyses of these benthic data indicate no
significant differences in the population structure prior to
plant operation.  Numeric densities for the various components
of the benthos are approximately the same.
     Following the plant startup and for all subsequent
collections, ecologically and statistically  significant
changes occur in the benthos.  Community structure is altered
and numeric differences occur.  These changes and differences
were found to be generally coincident with factors associated
with tailing discharge.  On the basis of these analyses, it
is possible to conclude that the release of tailings greatly
affects benthic organisms in Western Lake Superior.  This
effect should be understood with respect to the entire
ecosystem.
                             1313

-------
                      LITERATURE CITED
Adams, C. E., and R. D. Kregear.  1969.  Sedimentary and faunal
   environments of eastern Lake Superior.  Proc. llth Conf.
   Great Lakes Res.; Internat. Assoc. for Great Lakes Res.
   12:1-20.

Alley, W. P., 1968.  Ecology of the Burrowing amphipod
   Pontoporeia affinis in Lake Michigan.  Univ. Michigan,
   Great Lakes Res. Spec. Rep. No. 36, 131p.

     _, and R. F. Anderson.  1968.  Small-scale patterns of
   spatial distribution of the Lake Michigan macrobenthos.
   Proc. llth Conf. Great Lakes Res.; Internat. Assoc. for
   Great Lakes Res. 11:1-10.

Anderson, Emory D., and Lloyd L. Smith, Jr., 1971.  A synoptic
   study of food habits of 30 fish species from western
   Lake Superior.  Univ. Minn., Agr. Exper. Sta., Tech. Bui.
   279:199 pp.

Ayers, J. D., and J. C. K. Huang.  1967.  Studies of Milwaukee
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Baier, C. R., 1935.  Studien zur Hydrobakteriologic stehender
   Binnengewasser.  Archiv. f. Hydrobiol.  29:183-264.

Bousefield, E. L., 1958.  Fresh-water amphipod crustaceans of
   glaciated North America Canadian Field Nat.  72(2):55-113.

Bursa, A. S., and L. Johnson, 1967.  Nannoplankton of marine
   origin from Great Bear Lake in the Northwest Territories
   of Canada.  Nature 214(5087):528-529.

Carr, J. K., and J. K. Hiltunen.  1965.  Changes in the bottom
   fauna of western Lake Erie from 1930 to 1961.  Limnol.
   Oceanogr. 10:551-569.

Cook, G. W., and R. E. Powers.  1964.  The benthic fauna of Lake
   Michigan as affected by the St. Joseph River Proc. 7th Conf.
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   Rep. No. 11:68-76.

Daly, Reginald A., 1963.  The changing world of the ice age.
   Hafner, N. Y., 271 pp.

Eggleton, F. E.,  1936.  The deep-water bottom  fauna of Lake
   Michigan.  Pap. Mich. Ae
-------
Ekman, Sven, 1915.  Die Bodenfauna des Vattern, qualitativ und
   quantitativ Untersuchungen.  Int. Rev. Hydrobiol.  7:146-204;
   275-425.

Flint, R. F. , 1957.  Glacial and Pleistocene Geology.  Wiley, N.Y.

Goldstein, A., 1964.  Biostatistics.  Macmillan, N. Y., 272 pp.

Gordon, W. G. , 1961.  Food of the American smelt in Saginaw Bay,
   Lake Huron.  Trans. Amer. Fish Soc. 90:439-443.

Henson, E. B., 1954a.  Pontoporeia affinis var. brevicornis in
   Cayuga Lake, N. Y.  Ecology 35(4):579.

Henson, E. B., 1954b.  The profundal bottom fauna of Cayuga Lake.
   Unpubl. PhD thesis, Cornell Univ., 108 pp.

Henson, E. B., 1966.  A review of Great Lakes Benthos Research.
   Univ. Mich., Grt. Lakes Res. Div., Publ. 14:37-54.

Henson, E. B., 1970.  Pontoporeia affinis  (Crustacea, Amphipoda)
   in the Straits of Mackinac region.  Int. Assoc. Grt. Lakes
   Res., Proc. 13th Conf.:601-610.

Hiltunen, Jarl K., 1969.  Invertebrate macrobenthos of western
   Lake Superior.  Mich. Academician 1(3,4):123-133.

Hough, Jack L., 1958.  Geology of the Great Lakes.  Univ. 111.
   Press, Urbana, 313 pp.

Hubb, C. L., and K. F. Lagler.  1958.  Fishes of the Great Lakes
   Region.  Univ. of Michigan Press, Ann Arbor, Michigan.  213 p.

Kinney, W. L.  1972.  The macrobenthos of Lake Ontario.  Proc.
   15th Conf. Great Lakes Res. Internat. Assoc. for Great Lakes
   Res.  15:53-79.

Larkin, P. A.,  1948.  Pontoporeia and Mysis in Athabaska,
   Great Bear, and Great Slave Lakes.  Bull. Fish. Res. Bd.
   Canada, 78:1-33.

Lindstrom, G., 1855.  Bidrag till Kannendomen om Ostersjons
   Invertebrat-fauna.  Ofv. Kgl. Vet. Ak. Forh., Arg. 12.

Marzolf, G. R. , 1963.  Substrate relations of the burrowing
   amphipod Pontoporeia affinis Lindstrom.  PhD thesis,
   Univ. Michigan.  92 p.

Marzolf, G. Richard, 1965a.  Substrate relations of the bur-
   rowing amphipod Pontoporeia affinis in Lake Michigan.
   Ecology 46(5) :579-592.
                            1315

-------
Marzolf, G. Richard, 1965b.  Vertical migration of Pontoporeia
   affinis (Amphipoda)  in Lake Michigan.  Univ. Mich., Grt. Lakes
   Res. Div., Publ. 13:133-140.

McErlean, Andrew J., and Catherine Kerby, 1972.  Biota of
   Chesapeake Bay:  Introduction.  Chesapeake Sci. 13,
   Supplement:S4-S 7.

Moffett, James W., 1956.  Recent changes in the deep-water fish
   populations of Lake Michigan.  Trans. Amer. Fish. Soc.
   86:393-408.

Mozley, S. C., and W. P. Alley.  1973.  A comparative study of
   the Lake Superior, Lake Michigan, Lake Huron, and Lake Erie
   macrobenthos.  Bull. Fish. Bd. Canada.  Submitted for
   publication.

Muller, Otto, 1964.  Weichselzeitliche eisgestaute Seen als wesent-
   liche Elemente in der Ausbreitunggeschichte von marinen
   Glazialrelikten des Nordpolarmeeres.  Arch, fur Hydrobiol.,
   Erg. der Limnol.:1-90.

Nissen, 0., and P. 0. Ottestad, 1943.  On the analysis of variance
   and the effect of non-normality.  Meldingv fra Norges
   Landbruleshegskole:1-23.

Norton, A. H., 1909.  Some aquatic and terrestrial crustaceans
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   communities at various depths in Lake Champlain, Vermont
   (1967-68)  Unpubl. PhD Thesis, Dept. Zool., Univ. Vt., 110 pp.

Powers, C. F., and W. P. Alley.  1967.  Some preliminary observa-
   tions on the depth distribution of macrobenthos in Lake
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   No. 30:112-125.

Rawson, D. S.,  1951.  Studies of the fish of Great Slave Lake.
   J. Fish. Res. Bd. Canada.  8:207-240.

Ricker, K. E., 1959.  The origin of two glacial relict crustaceans
   in North America as related to Pleistocene glaciation.
   Canad. Jour. Zool.  37:817-893.

Robertson, A., and W. P. Alley.  1966.  A comparative study of
   Lake Michigan Macrobenthos.  Limnol. and Oceanog.  11:576-583.
                             1316

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Samter, M., and W. Weltner, 1904.  Biologische Eigentumlichkeiten
   der Mysis relicta, Pallasiella quadrispinosa und Pontoporeia
   affinis, erklart aus ihrer eiszeitlichen Entstehung.
   Zool. Anzeiger 27(22):676-694.

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   distribution and abundance of benthic fauna in Saginaw Bay,
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                             1317

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        0
Segerstrale, Sven G.,  1971b.   Further data on summer breeding
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   western basin and near-shore Canadian waters of Lake Erie.
   llth Conf.  Great Lakes Res.; Internat. Assoc. Great Lakes
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   Fish. Wild. Serv., 60:343-369.

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   Coregonus hoyi, in Lake Michigan.  Trans. Amer. Fish. Soc.
   92(3):245-255.

	., 1968.  Daytime distribution of Pontoporeia affinis
   off bottom  in Lake Michigan.  Limnol. Oceanogr.   13:703-705.


       ;                      1318

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Zobell, C. E., and C. B. Feltham.  1938.  Bacteria as food for
   certain marine invertebrates.  J. Mar. Res.  1:312-327.

     ,  and      .  1942.  The bacteria flora of a marine mud flat
   as an ecological factor.  Ecology  23:69-78.
                             1319

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Characterization of the North Shore Surface Waters of Lake Superior
                          Armond E. Lemke
           United States Environmental Protection Agency




                 National Water Quality Laboratory




                     Duluth, Minnesota  55804
                                1320 A

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                        Introduction






      Lake Superior, the largest body of fresh water in the world (in




surface area), was studied intensively along the north shore from Just




north of Two Harbors, Minnesota to Grand Marais, Minnesota.  This study




was conducted during the period from early July 1972 to late October




1972.  Tho study ^SjUlitiated to evaluate the water quality in the far




western end of Lake Superior, and to evaluate the impact on Lake Superior




of the daily dumping of 67,000 long tons of taconite tailings at Silver




Bay, Minnesota.  This study is prepared for litigation in the case of




United States vs. Reserve Mining Company, and consists of several phases.




This report characterizes the water conditions found in the water along




the North shore of Lake Superior with respect to the presence or absence of




taconite tailings, light penetration, temperature and bacteria, and phyto-




plankton differences in the euphotic zone.  Information gathered previously




by several investigators and reported in the hearings conducted by the




Federal Water Pollution Control Administration and the Federal Water




Quality Administration was the determining factor in deciding which




parameters received the most scrutiny.
                                   1321

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                          Station Locations




     The selection of the sampling sites for this investigation are




discussed in detail in a concurrent report by John W. Arthur.  Briefly,




selection was made to differentiate as much as possible those effects




caused by the addition of taconite tailings and those effects caused by




runoff from local streams along the north shore of Lake Superior.




Information from lake current studies, such as Adams (1970), indicated




that the general trend of the currents is from Silver Bay toward Duluth,




therefore, our stations were selected northeast of streams which enter




the lake along the North Shore.




     Most of the samples were taken from four transects established in




early July, and located as follows:  Silver Cliff approximately 5




miles northeast of Two Harbors, Minnesota; Split Rock Light House




approximately 7 miles southwest of Silver Bay, Minnesota; Shovel Point




(Crystal Bay Point) approximately 6 miles northeast of Silver Bay,




Minnesota; and Sugar Loaf Cove approximately 5 miles southwest of




Taconite Harbor, Minnesota.  At each of these locations the most




prominent radar landmark was selected and the stations were established




at one, three, and five nautical miles offshore, perpendicular to the




general trend of the shore line in each area.  The stations are




referred to by the first letter of each word in the station name and




a number for the offshore distance.  The station three miles offshore




at Split Rock Lighthouse will be called SR-3.  Buoys were placed at




each of these locations for use in another phase of the investigation




and samples were taken within one-eighth miles of the buoy.
                                1322

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     Location of these buoys during each sampling run was a major                 |


problem.  Summer conditions on Lake Superior are such, that during much           ;


of the calmest weather, the cold lake water causes surface fog, limiting          i

                                                                                  i
visibility.  The radar units used to initially locate the stations had            '
                                                                                  i
sufficient versatility so that we were able to get in the general area            I


of the buoy by vectoring off of the shore,  and then by changing range             j

                                                                                  i
actually locating the buoy.  Attrition of the surface buoys because of            !


severe weather during the last stages of the study was circumvented by            i
                                                                                  <
taking very careful vector readings from the shore by radar and then              '•


sampling in the area where these readings indicated the buoys had been.           \


On several occasions high waves obscured the buoy from direct radar               i
                                                                                  i
                                                                                  f
location and upon reaching the area indicated by the shore vectors the            '


buoy was within one-quarter mile by visual  observation, assuring us that          ;


samples taken without location of the buoy, were sufficiently  accurate           '

                                                                                  i
as to location.                                                                   i
                                                                                  ;
     Late in the study period two transects were established in the               j


Grand Marais, Minnesota area because taconite tailings were found at


all of the other transects.  These new stations were on transects


directly out from the Coast Guard station point at Grand Marais and


out from the large point northeast of Five Mile or Guano Rock,


approximately six and one-half miles northeast of Grand Marais.  See


map, Figure 1.
                               1323

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                         Sampling Techniques




     The kinds of samples and measurements made on site during this




work were as follows:  A six liter sample for analysis of total suspended




solids and tailings; A three gallon sample for plankton analysis which




was preserved immediately with Uttermohls solution; and a sterile




sample of at least 700 ml for bacteria analysis.  The first two were




taken with a Van Dorn water bottle of six liter capacity.  The bacteria




samples were taken with the use of a Zobell sampler modified to hold




a one liter sterile bottle.  This bottle was opened at depth by breaking




a small glass tube using a drop messenger which allows the sterilized




and evacuated bottle to partially fill with water and provides an




uncontaminated sample.  The bottle was retreived rapidly and the influent




tube clamped to maintain it in a uncontaminated condition.  Samples for




bacterial analyses were immediately iced.  Suspended solids-tailings




samples and bacteria samples were taken at forty and twenty foot depths




and the plankton analysis samples were taken only at the twenty foot




depth at each station.




     All samples were labelled on the bottle with a code number and depth




of collection.  A security seal and a chain of custody tag were affixed.




Some of the bottles used for the tailings-suspended solids samples were




self-sealing and no security seal was used on .these bottles.  All bottles




used for collecting samples were washed and rinsed with distilled water




prior to use.  These bottles were all new in the case of the algae




and tailings sample bottles.  The bacteria bottles were prepared in




the usual method by autoclaving after hot water washing and distilled
                                1324

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water rinsing.  More detailed descriptions of bottle preparation will

be found in the reports of Robert W. Andrew and Victor Cabelli for this

investigation.

     Site measurements included the following:  Secchi disc readings,

temperature readings each five feet from 45 ft to the surface and a

submarine photometer reading at each five feet.  In certain cases

during part of the study a current device, which consisted of a weighted

winged drogue supported by a light line and float of just sufficient size

to hold it at depth, was used to make rough measurements of current

direction and speed.  This data is reported in Table 4.  Some

directional measurements only were obtained by the diving crew working

on another part of the investigation.

     Subsequent to the retrieval of the last sample of the day's

operations, the samples were returned to a landing area by the most

direct route. If the area was different than that of a safe mooring,

the samples were returned to the National Water Quality Laboratory

by courier.  If the mooring and landing area were the same the

sampling crew returned the samples personally.

     The bacteria samples were processed at the National Water Quality
      •
Laboratory and a detailed discussion of the findings are in a concurrent

report by Victor Cabelli.-  1° this report only the total counts are

reported.  The samples for total suspended solids and tailings were

processed at the laboratory also and these methods are found in a

concurrent report by Robert W. Andrew.  The data reported here are from

those analyses.  Plankton samples were shipped to Dr. Alfred M. Beeton

at the University of Wisconsin, Milwaukee for analysis and his findings

will be reported separately.

                                 1325

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                         Discussion of Data




     The Secchi disc readings were used exclusively as a source of light




penetration determinations.  Secchi disc readings varied about 7 to 8




meters between the least clear and the most clear readings at each station.




Although the range was approximately the same the means were quite




different.  Ranging from 5.5 meters at Silver Cliff 1 mile to 10.9 meters




at Guano Rock 1 mile and Grand Marais 3 mile.  The maximum readings,




14.5 and 14 meters, were found at the Grand Marais and Guano Rock Stations,




respectively.  The lowest reading found during our regular sampling




was 2.5 meters at Silver Cliff 1 mile two days after the heavy rain of




September 20, 1972 on the adjacent shore area (Table 1).




     Total suspended solids data received from Robert W. Andrew and




appearing in Table 4 were plotted against Secchi disc for each station




after averaging the reported level for the twenty and forty foot samples




for each sampling date.  This was accomplished because the Secchi readings




are an integration of the light penetration down to the depth of the




reading, making the average of the two readings, a more realistic




estimation of the light scattering power at the sampling site.  The




plots appear in Figures 2-8 of this report.  The regression line and




the r indicated are calculated using all of the points indicated on the




graph but for clarity each of the stations is represented by a different




symbol.  Based on Snedecor and Cochran  (1967), the regression lines of




those plots at Silver Cliff, Split Rock are significant at the 1% level




and that of Shovel Point is significant at the 5% level.  Those at Sugar




Loaf, Grand Marais and Guano Rock are not significant at the 5% level.
                                1326

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                                                                     8






     Tailings concentrations as indicated by cummingtonite analysis and




expressed as mg/1 of tailings in the water samples were also received




from Robert W. Andrew.  Although tailings were measured in some of the




samples at each of the four stations closest to the point of discharge,




only those samples obtained at the Split Rock Stations had measurable




tailings in more than one-half of the samples taken.  Many samples for




which tailings were not measurable were associated with the plant shut-




down period.  A regression analysis of all samples taking Secchi disc vs.




tailing concentration yielded an r value of .212 which was significant at




5# but not at 1%.




      Fig. 2k shows a plot of Suspended Solids vs. tailings, taking all of




the 20 ft. samples in the study.  The r value for this plot is .358 which




is significant at 1%.  On this basis one can conclude that if there were




no tailings present the turbidity would be significantly lower.




     Plots of temperature vs. date of occurrence found in Figures 12-17




show that the maximum surface water temperatures in the study area




occurred during the last two weeks in August and reflect a period of




bright warm weather in an otherwise cold cloudy summer.  The shallow




sampling depths used precluded the detection of any thermocline.  The




five-mile forty-foot readings are the least variable, probably because




the greater wind action away from the land decreases the possibility




of local warm or cold spots.
                                1327

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              Overview of North Shore of Lake Superior




                      Surface Water Conditions






     Lake Superior is a classic example of an oligotrophic body of water.




Shallow water temperatures near shore reached an extreme of 15.5° C in




the study area.  The lake bottom depths at the study stations varied




from 450 feet at Guano Rock 1 mile to 900 feet at Split Rock 5 mile.




Total bacteria counts were 1000/100 ml or less except after heavy run-




off from adjacent land area.  Secchi disc readings were generally lower




southwest of Silver Bay and were extremely low during green water




episodes and storm run-off.




     Adams (1970) described the summer circulation patterns in Lake




Superior after a very comprehensive study over two seasons.  The limited




data which we developed supports his findings.  Of particular interest




are Adam's statements concerning upwelling along the North Shore




particularly under the influence of strong offshore winds.  Our sampling




of the Split Rock stations illustrated this very well.  On September 22




the 1 mile station Secchi was 5.0 meters, bacteria 29,100/100 ml, and




total suspended solids 1.15 mg/1.  At the three mile station the values




were 10.5, 1330, and 0.45, and at the five mile the values were 4.0,




44,000, and 1.45, respectively.  These readings obtained two days after




a rain of nearly 4 inches in 24 hours was recorded in the adjacent land




areas.  The values obtained at one and five miles were similar although




slightly higher than those obtained after a lesser rain the month previously




in the area.  The clear water readings at the three mile"station vere



apparently taken from a discreet body of water in that area.





                                  1328

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                                                                     10





     A detailed discussion of the bacterial flora at the sampling sites




on the North Shore will be presented in a concurrent report by Victor




Cabelli.  Data obtained from him and appearing in tabular form in his




report on the total counts were plotted against Secchi disc which were




shown previously to be related to solids in suspension (Figures 9-11) .




In all cases the r value reported is significant at the 1% level.




That such is the case has been reported by Pfister et al. (1968).  Any




increase in turbidity is likely to increase the bacterial flora in the




water.  Direct correlation of bacteria with total suspended solids




resulted in an r significant at 1%.




     Although our direction and current rate data is limited as




stated previously, an indication of the various currents which could be




encountered shown by the drogue data for Sugar Loaf transect for




October 4, 1972 (Table 4).  At one mile station the rate of travel was




12.5 ft/min and the direction was 210° or approximately southwest; three




mile rate was 10 ft/min and the direction was 130° or southeast; five




mile rate was 17 ft/min and the direction was 20° or north-northeast.




The wind was less than five miles per hour southwest this day.  This




suggests eddy currents on that date.  Many of the current direction values




obtained were in a south to west quadrant; that is the values ranged




between 180° and 270° magnetic.  At various times, however, all points




of the compass were represented.  This means that any material suspended




in the surface water of the lake can be transported in any direction.
                               1329

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                                                                     11






     Rainfall for the adjacent land area and the study area during the




sampling period was abnormally heavy.  The weather bureau at Duluth




Airport recorded a total of 3.82 inches of rain on August 15-16, 3.95




inches on August 20-21 and nearly a one day record of 3.77 inches




on September 20.  Secchi disc, total suspended solids and total bacteria




counts all show the effect of the land run-off from this precipitation.




The rains in July were more general along the shore than those in August.




All stations showed the effects, with the 1 mile stations being most




affected, except at Split Rock where the 3 mile station shows the




most effect.  Secchi values of 4-5 meters, total suspended solids near




2 mg/1 and bacteria counts above 10,000/100 ml were found at three of




the stations.  The August 20 rain was concentrated from Silver Bay toward




Duluth and at the Silver Cliff stations the lowest Secchi values 2.5-3.5




meters, highest suspended solids 1.3-2.0 mg/1 and highest total bacteria




counts 24,600/100 ml to 35,000/100 ml of the study period at regular




stations were observed.
                                1330

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                                                                     12
     Observations of Effects Attributable to Taconite Operations





     Visual effects of the taconite operations, i.e., green water was




an intermittent phenomenon and will be handled separately in this




report.  A plot of the tailings concentration at all of the stations




vs. date are shown in Figures 18-22.  Also shown on these Figures




are the dates of no operation of the taconite processing plant.  Those




samples obtained prior to plant shut down all contained taconite




tailings.  After shutdown less and less tailings were found in the




area until in the samplings accomplished on August 28 and 29 tailings




were not measurable.  At the next sampling period> approximately two




and one-half weeks after plant start up, tailings again were measured




in the samples and by the end of the sampling period all stations




had shown tailings at least once.  The most rapid disappearance and




reappearance of tailings after plant shutdown and start up occurred




at the Split Rock stations which were closest to the discharge area.
                                1331

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                                                                     13





                             Green Water






     Green water along the north shore of Lake Superior received study




during the sampling period.  Two rather large green water masses were




sampled to check the amount of tailings occurring as dissolved solids




in the green water areas.  Samples taken in areas affected by the heavy




run-off mentioned previously were also compared.




     The first striking episode of green water was noted on September 1,




1972, approximately one week after start up of the taconite operation




following a one month shutdown.  :  Meterological conditions were




a clearing sky and a strong northwest wind in the plant area.  Samples




taken near the edge of the green water area had total solids of 2.0 and




1.1 mg/1 and tailings concentrations of 2.0 and 0.7 mg/1 at 20 and 40 ft,




respectively.  At the time of the sampling the green water area extended




from the delta to southeast of King's Landing but was discontinuous in




the area of the SR-1 station.  A sample taken there at the 20 ft level




contained 1.0 mg/1 suspended solids and 0.7 mg/1 tailings with a Secchi




disc reading of 6.0 meters.  A sample taken 1 mile south of Pellet




Island contained 3.4 mg/1 suspended solids, of which at least 2.5 mg/1




was tailings.




     The second green water episode was first observed early on October 17,




1972.  The only measurable precipitation reported for October 1-19 was




0.29 inches on October 10; thus North Shore streams were observed to be




low and clear on October 17.  Green water samples were taken in the




afternoon of October 17.  At this time the green water was discontinuous




from the delta of the taconite processing operation, but extended in a
                               1332

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                                                                     14

solid mass from Pellet Island in an approximately one-half mile wide band

to Split Rock light.  Southwest from there it fanned out into the lake so

that at the mouth of the Gooseberry River it was about 7 miles wide.

The green water became discontinuous in the vicinity of Encampment Island

(Figure 23).

     Four sets of samples, each set consisting of a sample from 5 ft and

40 ft, were taken at this time.  One set was obtained in a clear blue

water area approximately one mile directly off shore from the delta.  A

second set also in an apparent clear blue area was taken approximately

1 mile directly offshore from King's Landing.  The third set was obtained

from a green water area approximately one-fourth mile off King's Landing.

The fourth area selected from the air as  the most green area, was

sampled approximately one-third mile southwest of Split Rock Point.

Table 2 presents the values found at these sampling sites.  Temperature

data (Table 3) indicates that the clear water directly in front of the

delta was water inside the harbor and was colder than that about-a mile

offshore.  The values of Table 2 appear to substantiate this cold area

as an upwell because of the low suspended solids and high Secchi readings.

Tailing amounts were the lowest in the very clear surface water adjacent

to the delta.  Next lowest tailings were found in the apparent blue water

in sample group two.  The highest levels were found in the area of heaviest

green water and are the highest tailings, total suspended solid

concentrations, and lowest Secchi readings found in the open lake during

the study.  Of interest is the difference in Secchi disc determinations

between set 1 and set 2 where the total suspended solids are approximately

equal, but the amount contributed by the tailings is much more, and the

Secchi and concurrent light penetration is reduced.
                              1333

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                                                                     15






     Apparent color is different, in areas with low Secchi readings




(15-3.5 meters) and high suspended solids (1.5-3.0 mg/J), if the tailings




fractions of the total solids differ markedly.  The color of the high




tailings area is a bright almost chartruse green, whereas the apparent




color after heavy run-off, as noted on the August 25 and September 22




sampling runs, was a more brownish to grey color.  The run-off water




was accompanied by large amounts of floating debris not found in the




green water high taconite areas.
                               1334

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                                                                    16






                             Conclusions





     As a result of the described studies and personal observations,




the following conclusions have been formulated by this author.




     The taconite processing operations at Silver Bay, Minnesota are




contributing to an increase in turbidity in Lake Superior, particularly




in the area southwest of Silver Bay, but also to a lesser extent in




other nearby areas.  Suspended solids in the same area have been




increased as the result of the ore-processing operations.




     Material from the tailings discharge is being transported  to all




parts of the study area, and current data indicates that this being




the case the tailings fines are at least being distributed throughout




the Duluth arm of Lake Superior.




     The green water seen along the North Shore of Lake Superior is




being caused in a significant part of the episodes by particulate




matter from the tailings operations.
                               1335

-------
                            Bibliography





Adams, Charles E. Jr.  Summer circulation in Western Lr>'<-e Superior.




     Great Lakes Res. Center, U. S. Lake Survey, Detroit, Mich. 1970.




Pfister, Robert M., Patrick R. Dugan, and James I. Frea.  Particulate




     fractions in water and the relationship to aquatic microflora.




     Proc. llth Conf. Great Lakes Res., pp. 111-116.  1968.
                                 1336

-------
Table 1.  Secchi disc mean maximum and minimum in meters.
Station
Silver Cliff
1
3
5
Split Rock
1
3
5
Shovel Point
1
3
5
Sugar Loaf
1
3
5
Grand Marais
1
3
5
Guano Rock
1
3
5
Mean

5.5
6.8
7.2

7.2
7.7
7.0

9.0
9.0
8.7

8.85
9.75
9.70

8.75
10.9
9.3

10.9
10.5
10.3
Maximum

7.5
9.5
9.0

11.5
10.5
9.0

11.5
13.5
12.0

12.0
13.5
12.5

11.5
14.6
11.0

14.0
13.0
12.0
Minimum

2.5
3.5
3.5

4.0
4.5
4.0

7.0
6.5
5.0

5.0
7.0
7.5

7.0
8.5
7.5

7.0
8.0
9.0
                         1337

-------
                                                                 JJ
            Table 2.  Results of analysis of green water samples.
             (a)
                                           Total suspended
          Location            Secchi (m)   solids rug/liter   Tailiu6s mg/liter
9/1/72

  2.75 mi. SW Reserve delta
    20'                           3.0            2.0                2.0
    40'                           3.0            1.1                0.7

  1 mi. South Pellet Island
    20'                           2.5            3.4               +2.5

9/1/72

  In discontinuous area
   Split Rock 1 Buoy
    20'                           5.0            1.0 '               0.7

9/27/72

  Greenwater patch
   Cove: North Split Rock Light.
    Surface sample                6.0            0.9                0.9
10/17/72

  1 mile offshore King !sLanding
    Clear area
      5'                          6.5             .7                 .1
     40'                                          .3                 .2

  1 mile southeast Reserve Harbor
    Clear area
      5'                         11.0             .4               < .1
     40'                                          .5               < .1

  1/4 mi. offshore King's Landing
    Green area
      5'                          1.8            2.2                2.0
     40'                                         1.9                1.7

1/3 mi. southwest Split Rock Point
  Heavy Green
    5'                            1.4            5.4                4.8
   40'                                           4.4                4.0
      All data provisional pending establishment of accuracy of methods of
      analysis.                      1338

-------
             Table 2.  Temperature on October 17, 1972.
              (b)
Location
1 1/4 mile 120* from
Reserve Harbor
1 mile
3/4 mile
1/2 mile
1/4 mile
Harbor mouth
In harbor
Surface 5 ft. 40 ft.
5.1° C
4.9
4.8
4.6
4.5
4.4
4.4
1 mile off King's Landing         4.2*            4.0*           4.0*
 and
4 miles from Pellet Island
 out of green water

1/3 mile off King's Landing       4.0*            3.9*           3.5*
 and        ,
4 miles from Pellet Island
 in green water

1/3 mile off Split Rock Point     4.0*            3.2*           3.0*
 in very green water
        Temperatures not calibrated - all temperatures relative.
                              1339

-------

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                                           1377

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       The Effects of Taconite Tailings
                    on the
        Phytoplankton of Lake Superior
                   May 1973


                Joseph Shapiro
                  1378 A
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                          INTRODUCTION

     During th« past few years considerable interest has developed

in d«terminin$ the chemical factors that limit and control the

growth of planktonic algae in Lake Superior.  This interest stems

both from g«n»ral awareness of the problem of eutrophication, or

over-enrichflWint of lakes, that has become prominent in recent years,

and from th« us* of Lake Superior as a receiving body for taconite

tailings.  Various studies have been done, aimed mostly at determining

the limiting factors and finding out whether taconite itself stimulates
                                          i
algal growth.  Among the latter studies are those by Andrew and Glass

(1970), Goldman  (1970), McGee (1970), and Shapiro  (1970).  The results

of thes« inv««tigations, although individually too few to permit

statistical evaluation, together provide consistent evidence that

taconite tailings do stimulate the growth of the native algae in

Lake Superior.  Furthermore, studies by Schelske et. al.  (1972) and

by Shapiro and Glass (1970) have established that the algae of the

open waters of Lake Superior are limited by phosphorus and manganese.

     It was in an attempt to determine the validity of these results

and to clarify any relationships between the stimulation by taconite

and by phosphorus and manganese that the present study was undertaken.
                            1379

-------
                            METHODS





     The experiments described in this report were carried out ii



the laboratories of the Limnological Research Center, University



of Minnesota, Minneapolis, under the immediate and continuous



supervision of Dr. Joseph Shapiro, Professor of Ecology and



Associate Director of the Limnological Research Center.  All were



done on samples of water collected from Lake Superior by the staff



of the National Water Quality Laboratory at Duluth, and shipped in



iced containers directly to the Limnological Research Center by



truck.  Samples of taconite tailings were shipped at the same time.



     All experiments were similar in design and execution.  Samples



(800 ml) of the lake water containing the native algae were incubate



in screw-capped 1 liter Erlenmeyer flasks in a light incubator at a



light intensity of from 200 to 400 foot candles and temperatures



from 6 to 8°C.  To some of the flasks were added various amounts of



taconite tailings and to others various amounts of phosphate, and/or



manganese.  Some of the flasks were wrapped in heavy aluminum foil



to act as dark controls.



     Evaluation of the effects of the treatments on the growth of



the algae was done on samples taken periodically and by use of three



different procedures,o~14 uptake, chlorophyll fluorescence, and



algal counting.





1. C~ 14  up take



     One method for determining the response of the algae  to the



treatments would be to filter them from the water and weigh them.



However, the  relatively low concentration of algae in  Lake Superior



waters makes  it impossible to perform this procedure with  any



degree of accuracy and so a "tracer"technique  was used.   When the



water was first received, and before it was poured into the  flasks,





                           1380

-------
one milliCurie' of radio-carbon, C-14f wag added as NaHCO3 to about




35 liters of it.  The very small amount of sodium bicarbonate



added little to the carbon content of the water, and did not change



the pH, but it did enable the incorporation of carbon into algal



material to be followed by periodically filtering samples of algae



from the flasks and determining the degree of radioactivity of



those algae.  Specifically, water samples containing 50 ml were



filtered through a 1 inch diameter 0.45 micron Millipore filter,



the filter was glued to an aluminum pUanchette, dried, and the



radioactivity counted in a Picker Proportional Counter,



     Several special precautions were taken to insure accuracy:



ja. In the first experiment, only 15 inches of vacuum were used to



draw the samples through the filters so as not to damage the algae.



Subsequent experiments on a single flask showed (table below) that a



full vacuum of 30 inches could be safely used and so this was done



in subsequent experiments.







          Vacuum used                   cpm on filter



              15"                           7167



              15"                           6966



              15"                           6869



              30"                           6684



              30"                           6639



              30"                           7065





b.  After drawing the samples through the filters, double distilled



water was drawn through them to rinse excess radioactivity from



the filters before removing them from the apparatus.  To verify the



utility of this procedure, experiments were done in which two fil-



ters were used in series and the radioactivity of the lower one






                           1381

-------
determined separately.  As shown below, the second filter contained



almost no radioactivity indicating that the rinsing procedure was



effective.





               Filter                   cpm
                top                     1727



                bottom                     9





                top                     1017



                bottom                     5






c_.  Dark controls were used to verify that C-14 "uptake" was indeed



uptake by photosynthesizing algae rather than adsorption onto



chemical precipitates.






d.  Sufficient counts were counted in each case for statistical



accuracy, i.e, 10,000-20,000, or 5 minutes.






     During a previous investigation along the same lines this



approach to monitoring algal growth was criticized on the ground



that it used long-term uptake of C-14 by the algae instead of the



2-4 hours recommended in several publications (eg. Vollenweider,



1969).  It should be made clear that the 2-4 hour periods were



recommended for measuring instantaneous rates of photosynthesis but



that what is being measured here is growth, and, in order to measure



growth, it is necessary to allow the algae to accumulate carbon for



sufficiently long periods of time to determine whether the growth



is indeed different in different circumstances.  In fact rather



than the long-term uptake of C-14 invalidating the results, it tend



to reinforce them.  As the algae grow, some of the C-14 already



incorporated must be  respired and returned to the system.  Thus the



amount of C-14 held by the algae will decrease.  As this process is



                           1382

-------
likely to be faster in more rapidly growing algae, the long exposure



would tend to flatten out any response so that any differences



found would, in fact, tend to be minimized.  Thus if any error is



present it will result in the underestimation of differences between



treatments rather than emphasize them.







2.  Chlorophyll



     The second method used for evaluation of algal response was to



determine the fluorescence of the chlorophyll of the algae by means



of a fluorometer.  The instrument used was a Turner Model III self-



balancing Fluorometer modified with the appropriate photomultiplier



and filters as recommended by Lorenzen.  All samples were read in



a large volume, flow-through cell at lOx sensitivity.  Samples con-



taining enough chlorophyll to send the instrument off-scale were



diluted with double distilled deionized water and their fluorescence




calculated.  The instrument used has been shown to be linear in



response throughout the range of measurements made.  Because the



higher concentrations of taconite added fluorescence to the samples



the graphs are presented as changes from the original values.








3.  Algal numbers



     In order to verify that changes in radioactivity or chlorophyll



really did reflect changes in algal abundance, the algal populations



in certain flasks were examined and enumerated at the beginning and



end of each experiment.  Samples preserved in Lugol's solution were



settled in counting chambers and examined using a Wild inverted



microscope.  All species present and their numbers were enumerated.
                           1383

-------
                            RESULTS





     Because the work involved three samples of Lake Superior



water collected at different times and because the three samples



were used for somewhat different purposes, the results are



presented as three separate experiments.
Experiment I




     The water with which this experiment was done was collected



August 17, 1972 at 2:15 PM, 5 1/2 miles off shore at Sugar Loaf CPve



The experiment was set up between noon and 5 PM August 18 and con-



tinued to the afternoon of September 6.  The light intensity at



the center of the flasks was 400 foot candles to begin with, later



reduced to 200 foot candles and the temperature in the cabinet was



7-8°C.



     The purposes of the experiment were twofold;



1.  To determine whether or not taconite tailings stimulate algal



growth.



2.   To determine whether the addition of manganese stimulates algal



growth.  This was done because the previous work had suggested that



Lake Superior algae were manganese limited and that stimulation



previously shown by tailings might have been due to the manganese



leaching from them,



     Taconite tailings were added to the appropriate flasks, as a



slurry, at volumes relative to the lake water of .0001%,  .001%,



.01%, and  .10%.  This produced concentrations by weight of 0.41 mg/1



4.1 mg/1,  41 mg/1, and  410 mg/1.  Manganese was added  as  a solution



of MnCl2*4H?0 in volumes sufficient to produce concentrations of



Mn of 2, 4, 8, and 16 micrograms/liter.  Where possible replicates
                            1384

-------
and dark controls were used, as shown in Tables 1,2, and 3, and




Figures 1, 2, 3, and 4.








                  EFFECTS OF TACONITE ALONE




014



     The stimulatory influence of taconite tailings on C-14 uptake



is clearly shown in Table 1 and Figure 1.  While the addition of



.0001% tailings had no positive effect,  (but see Figs. 5 and 11),



concentrations of .001% and .01% clearly stimulated the algae to



take up C-14 at considerably increased rates over the controls.



Filtration of 0.1% suspensions was done only on the last day due to



the great difficulty of filtering them.  These showed inhibition,



possibly because the high turbidity of the suspensions inhibited



photosynthesis.



     Note that although some differences occur among the replicates



e.g. .001% C  (day 19) and .0001% B (day  19),'     in general;e,g.



.01% day 19, replicates agree very well.  Note too that the uptake



of C-14 in the dark controls was negligible showing that the high



counts were most likely the result of photosynthetic activity by



the algae.  Statistical analysis  (Keller) shows a highly significant



linear relationship between C-14 uptake  and taconite concentration



in the range  .0001-,01% taconite  (r=0.86, P=0.0006).





Chlorophyll




     Table 2 and Figure 2 illustrate the effect of taconite tailings



on the production of chlorophyll in the  flasks.  These results,



although less regular, corroborate the C-14 findings and show that



on the day of termination of the experiment the response was positive



at all concentrations of taconite from .0001% to 0.1%.  The initial
                            1385

-------
decline in chlorophyll values visible on the 4th day wa'3 attributed



to too intense illumination and the lighting was reduced by half



on day 5.  Again the replicates were generally close in value.






Algal counts



     Three samples were enumerated  (Table 3}, the original water



sample, one of the controls at day  19, and one of the  .001% raconite



flasks at day 19.  The last was done instead of the  .01% sample



which had shown greater stimulation, but in which the  taconite



particles interfered with counting.  The results confirmed that



more algae had grown in the flasks  containing taconite.  In fact  the



ratio of the numbers of algae in the .001% flasks to the control,



2517/1304 = 1.9, is close to the ratio of the C-14 counts in  the



same two flasks  (1.6).  The pattern of response is interesting.



Among the algae most stimulated to  grow by the taconite were  Fragilai




Asterionella, small Cyclotella and  Stephanodiscus, Dinobryon  sociale,



anc^ Di^dymocystis.  Statistical analysis  (Keller) shows that the



relative abundance of the     major groups differs significantly  be-



tween the control flask and the .001% taconite flask,  and between the



control population  and both final  populations.





Conclusion



     The conclusion is clear.  By all three criteria,  C-14 Uptake,



chlorophyll increase, increase in algal  numbers, the taconite stimula



growth by the algae present in the  lake.








                   EFFECTS OF MANGANESE  ALONE



     Tables 1 and 2 and Figures 3 and 4  summarize the  effects of




manganese additions.
                            1386

-------
                                10
C-14 uptake


     In every case, by the 19th day the average counts in the flasks


with manganese added exceeded those for the controls.  However the


effects are, even allowing for some discrepancy in. the replicates,


not linear.  Four micrograms/liter of manganese clearly is more


stimulatory than 8 or 16 and more so than 2.  This type of response


has been noted by the investigat or and others  (e.g. Goldman 1966)


for trace metal   stimulation of algae.
                                    v


Chlorophyll

                                      I
     These data (Figure 4), again showing the effects of too much


light to the fourth day, corroborate the greater stimulation of  the


lower concentrations of manganese.



Conclusion


     The algae in the sample of water with which the experiment  was


done responded positively to small additions of manganese in the


range 2-4 microgram/liter but were inhibited at concentrations of


8-16 micrograms/liter,



Discussion


     The data above show that the algae were stimulated by tailings


and by manganese.  Because tailings have been shown to leach manganese


(Glass 1970) the question arises as to whether  this is the mechanism


by which taconite stimulates the algae, and whether it is the sole


mechanism.  Comparison of the C-14 uptake by the .01% flask  (Figure


1) with the C-14 uptake by the manganese-containing flasks  (Figure 3)


suggests that no manganese addition can stimulate  the algae as much


as can taconite alone f  i.e.  that  taconite  contains something  that


 stimulates  algae beyond what  can  be  achieved by manganese.   However,


 aa addition of manganese  to  taconite  causes no  further  stimulation


                            1387

-------
( i'lcy^es  1  =m-."> L;  Tables  '  ?J-d  _

that dt  least part of  uiie  cscry^:.
                                                         re f  it  • si evlv-vii "

                                                         . s ,  in  "act,,  due
      jhjs the  oc'.'iO j us: 3r

that are  now  limit i:icr  th

subs tonnes 1.;.  ..langan-^e.

l.-r er.
                                               o
                                                         1 J r. ?  tW',

                                                          -ad  L'l
Acce.s s :)j. v :-j tudios

      Because  this  v;as  the  f.i r,  t

ca I :,viat ioas  wore.-:  made to  ^a-.-clat

1...   Do-^a Lhe addition of  '"- i -i--1 ",
inci 3SS",? . he  alkalinity of t if. v •.
i-.l:as the  supply  of CO^?  The oh • -;

fsed v/.?s  d -•*. r or?1?' riecl by t j ttv^ <•.-C!>

r-^coided ^-i1.!! a  Bec;:",-.','i }-n RIO L-.. / .

of  0,57  a-'d O.^f, mM/1  r-;-3,'--icLi\",i :

1  mi,  pro"->.  a  viai  COD !r,tj.;iin'_; J i>i  ' ; .' .

rl  wit'vr, to  a volume  r> ' a'^'. "• -. • i n-' . ''.y

""'hat jr.,  '/, L  i-i liiCurio-   i r. ,,,','• .'.

t'"-.  i    -. i

iini llj T;T/1'VG /lire c ,   This  ;:--. i-c _;•  • n..--  \

detect i;;"'") by  vhe inethod  ?,s,-J,   'h.i-



ond 0.70, v,jluf'"3 iiirobabl-  r.r :  : ; .-.; i'-c

the un3piked  san'r->]i'-:  (0 67  anh  0 ,'-•  v>

not cause more tuan  a  ri,^;vL  i'-^:r"-   .

2.)   Does 'c.h~± aduition ^f  ; '^;r?'r  • :\ :

£ollo\>": r:cr tioie  ^urpmarizes a-it a  L.U   "' .
                                          -.-. •.•.cr.t:  b^vr\ra'.  irioe.: are^er is  j.nd.

                                           c-.- r '".  :.'.  -•i.'.-.~ inpvionci .

                                            . ' ~ n  Jr. i---;-:.:•-n^te in a ' eri -j.i ly
                                          -v: .../^"
                                            •   •  .,,f  ':.-.   -ke Mu -',-v.i.or ^at;Nr

                                           : r  , n,.   r.  IL,-,'^.',  f;c  .^ nil  of  4 , „= ,:i.
                                                       /.   H

                                           •' -'j  j- L: i'-  '.--C..ODS  vaejded  vax.ie'-:,

                                                    ^ :>:. of the  C-14  was  as

                                                    : t 1  •, " I'.jvar Mal!CO../: -, J

                                                     : ! 0- r-  o'- ? ake water „

                                                     '-•/•"!• JO,  v/3P  'tsedv   Ti'.i.-o

                                                    r-. ,   " , ": 3  rrd lii ,-.ranu:/35 ^ i tf-

                                                    .-:'3 i  '- ;  IfSK1  ~;''an  0,001

                                                    •  .  rv:r',;,.;so  and is i>.?low

                                                   >;. rob ;r--1 ed  by  titrati on  of

                                                    : .  .  'I...-: inil-ies uf  C^ (>9
                                                            • j '7j.t],  C--14  doe,

                                                    i ^ ' 1 i "^ \ "" V

-------
                                12
                                    Alkalinity raM/1.
Sample                          Day 0             Day 19
Lake Superior unspiked          .67, .68
Lake Superior spiked            .69, .70          .71
Lake Superior spiked + .0001%   .69, .70
Lake Superior spiked + .001%    .69, .70
Lake Superior spiked + .01%     .71, .71          .73
Lake Superior spiked + .10%     .81, .81       i   .85

Addition of taconite does indeed increase the alkalinity but the
effect is detectable only at concentrations of taconite exceeding
.01% by volume.  As even .001% caused algal growth stimulation
the stimulatory effect was not likely due to increased availability
of C0».  It is possible that a small part of the depressant effect
of 0.1% taconite on C-14 uptake is due to the increased alkalinity
caused by this addition as this would dilute the C-14 of the spike
and suggest an apparently lowered uptake.
3.)  Did the alkalinity decrease in the sealed flasks during the
experiment i. e. did using screw-capped flasks affect the results?
The data in the.above table for day 19 show  that this did not
happen.  The amount of growth that took place used only an insig-
nificant fraction of the CO,, available.


Experiment II
     This experiment, begun at noon on September 25 and extending
to October 16, was done with water collected on September 23, seven
miles off shore, six miles NE of Hovland, Minnesota and stored in
the dark at low temperature.  Illumination in the incubator was
                           1389

-------
                                13
maintained at about 200 foot candles and the temperature was



about 7°C.  C-14 spike was as in experiment I.



     The purposes of this experiment were?



1.  To repeat the taconite stimulation portion-of the first



experiment



2.  To determine the effects of phosphate addition to Lake Superior



water



3.  To determine the effects of adding phosphate and taconite



together, with and without manganese.



     The freshly supplied taconite slurry was  added at three con-



centrations -- .0001%, .001%, .01%.  Manganese was added as before,



as MnCl^, and phosphate was added as KH2P04.   The results are



shown in Tables 4, 5, and 6, and Figures 5-10.








                   EFFECTS OF TACONITE ALONE






C-14



     The addition of taconite alone stimulated C-14 uptake by the



algae at all concentrations used  (Figure 5),   These results corroborc



those of experiment I and show that even the lowest level of taconite



used, .0001%, has a stimulatory effect.  As in experiment I the



replicates agree well and dark controls showed negligible uptake of



the isotope.  Statistical Analysis  (Keller) shows a highly significar



linear relationship between C-14 uptake and taconite concentration ir




the range  .0001-.01% taconite  (r=0.91, P=,0001).






Chlorophyll



     The  chlorophyll data  (Table  5  and Figure  6) are irregular, but



tend to  substantiate the C-14 data.
                            1390

-------
                                14
Algal counts


     The first three columns in Table 6 show the results of algal


counts at day 0, and after 21 days on one of the controls and on


one of the .001% taconite flasks.  Again it would have shown more


effect had the algae at the .01% taconite concentration been


counted, but the particles of taconite once more interfered with


the observation of the algae under the microscope.  As in experiment


I these results corroborate the C-14 data.  The total number of


algae in the control at day 21 was about 1500/ml and that in the


taconite flask 2000/ml.  Again Fragilaria, Stephanodiscus and
                                                               t

Cyclotella and Dinobryon sociale provided much of the increase.


Statistical analysis (Keller)  shows that the distribution of algae


among the four major groups was different from the control in the


presence of taconite.



Conclusion


     Taconite tailings stimulate algae growth in Lake Superior as


measured by all three criteria.





                   EFFECT OF PHOSPHATE ALONE


C-14


     Table 4 and Figure 7  (note scale) show the striking effect of


phosphate addition to the Lake Superior water.  Except for the last


day, 10 micrograms P/liter seemed to be even more stimulatory than


20 — similar to the manganese effect in experiment I.



Chlorophyll


     The chlorophyll results in Figure 8  (note scale) corroborate


the phosphorus stimulation, again with 10 micrograms P/liter being


at least equal in effect to 20 except for the last sampling.



                            1391

-------
                                15
Algae



     The algae counts (Columns 5 and 6, Table 6) show that both



10 and 20 micrograms B/liter resulted in significantly more algal



growth than in the control, especially in the case of the 10



micrograms P/l where three algae, Fragi1aria, Osci1iatoria, and



Didymocystis grew exceptionally well.  Statistical analysis (Keller)



shows that the populations in the P-containing  flasks were different



from that in the control.






Conclusion



     Addition of phosphate alone stimulates algal growth in Lake



Superior water.








               EFFECTS OF TACONITE PLUS PHOSPHATE





     Because taconite seemed in Experiment I to provide two sources



of algal stimulation, one of which was probably manganese, and



because phosphate alone  can stimulate algal growth, combinations of



taconite and phosphate with and without manganese were used to



determine whether the second stimulatory agent  of taconite could in



fact be phosphate.






C-14



     The experiments performed are detailed in  Table  4 along with



the results as plotted in Figures 5,7, and 9.   Again  adding manganes



to .001% taconite caused no further  stimulation (Figure 5) but  again



taconite was capable of  greater  stimulation  (.01%, Figure  5) than




was manganese  (Fig.  7).  The greatest effects  (Figure 9) were



exhibited however when 20 micrograms of phosphorus were added to



 .001%  taconite and  to  .01% taconite.  C-14 uptake stimulation was



very high in both cases  and clearly  exceeded that due to taconite




                            1392

-------
                                16
alone at any level or to phosphate at the 10 and 20 microgram levels

of addition i.e. the effect was markedly synergistic.  Interestingly,

the flasks to which were added 20 micrograms P plus 8 rnicrograms

Mn, while showing great stimulation, yielded significantly lower

results, again suggesting that taconite supplies something, though

perhaps not phosphate, in addition to Mn.


Chlorophyll

     Table 5 and Figure 10 show, as with C-14 uptake, that in the

presence of either .001 or .01% taconite, 20 micrograms/liter of

phosphorus cause great stimulation, greater than 20 micrograms

phosphorus alone or than 20 micrograms phosphorus and 8 micrograms

manganese.


Algal counts

     The same effects of phosphorus on taconite are seen in Table 6,

columns 3 and 4.  The flask with .001% taconite alone had about

2000 algal cells/ml after 21 days but the addition of 20 micrograms

of phosphorus increased this to 76,500 — about 38-fold,  Interestingly,

using this mode of evaluation the substitution of 8 micrograms of

manganese for the taconite resulted in about the same growth of the
 algae  (Column  8} .  The taconite and manga,nese both stimulated the same
algae in the presence of phosphate — Fragilaria, Asterione11a,

Stephanodiscus  and Cyclotella, Didymocystis, and possibly Oscillatoria.

Although statistical analysis  (Keller) suggests that the populations

in the  two flasks, .001% taconite + 20ppbP>and SppbMn + 20ppbP5are

qualitatively different, inspection of the two sets of algal counts

suggests that they are remarkably similar, qualitatively as well as

quantitatively.   This is indicated in the table, below in which the

numbers of algae  in the two flasks are shown in relation to the

numbers at the  beginning of the experiment.  This great similarity


                            1393

-------
                                17
is further evidence that it is the manganese of the taconite vhnt
is its primary algal-stimulating component.  The statistical analyst
must be looked at here in the light of the lack of replications, the
high variability in algal enumeration, and the fact that the test
used, Chi-squared, is particularly sensitive to small differences.

                            Increase with           Increase with
   Algal Group              . 001%T + 20P             8Mn___+ 2OP
     Chrysophyta              116 fold                 171 fold
     Cyanophyta                68 fold                 100 fold
     Chlorophyta               90 fold                  73 fold
     Cryptomonads               '0.52 fold                0.38 fold
      and flagellatea
     Total                     70 fold                  80 fold
                                                           »
Conclusion
     The growth stimulating properties of taconite are greatly
enhanced by the presence of phosphate.  Probably the taconite
supplies sufficient manganese so that the algae are capable of
using the phosphate.  In the presence of 20 micrograms/liter of
phosphorus^ .001% taconite and .01% taconite are about equal in stimu-
lation.  It would appear therefore as though .001% taconite yielded
enough manganese for all the phosphate to be used.  Comparison with
the experiment with 8 micrograms of manganese plus 20 micrograms of
phosphorus suggests that the  .001% taconite is providing more than
8 micrograms of manganese/liter, or that the taconite has a growth
stimulator in addition to manganese and not identical with phosphate,

Experiment III
     This experiment.   was done  from November 2 to November 22 with
water collected November 1 offshore at Grand Portage Bay.  This samp;

                            1394

-------
was closer inshore than the previous two.  The incubator illumina-




tion was about 200 foot-candles, temperature 6°C.  The C-14 spike



used in this experiment was half that of the previous two experiments



due to a shortage of the isotope.  The purposes of this experiment



were:



1.  To test the stimulatory effect of taconite alone,



2.  To test the stimulatory effect of manganese alone,



3.  To test the stimulatory effect of taconite at two levels of




phosphate.








                   EFFECTS OF TACONITE ALONE





C-14



     Table 7 shows the experiments done and the results as  graphed



in Figure 11.  In this experiment the concentrations of taconite



used were lower, ranging from .00001% to ,01%,  Because of  the



many combinations used, replicates were fewer, as were dark controls.



However again increasing concentrations of taconite seeing to stimulate



the algae to a greater extent.  The stimulation seems less  than in



Experiments I and II, possibly  because the sample had been  contaminated



with manganese from  land runoff, but, as in the other two experiments,



statistical analysis  (Keller) shows a highly significant correlation



between C-14 uptake  and taconite concentration in the range .00001%-



.01%  (r=0.92, P=0.0008).






Chlorophyll



     Chlorophyll data, Table  8  and Figure  12, substantiate  the



stimulatory effects  of the taconite.






Algal counts




     Columns 1, 2, and 3 in Table 9 corroborate the growth  stimulatory




                            1395

-------
                                19
effect of the taconite.  Again it was not possible to enumerate the



algae in the flasks showing the most stimulation (.01%T).






Algal identification



     According to Ruth Beeton who examined the preserved samples,



the great increase in the Stephanodiscus-Cyclotella group was



mostly due to SteBhanodiscus Jhajntzschii  (may be S. tentfis^ but more



likely hantzschii) with some contribution from Cyclptella pee11ata^






Conclusion



     Although the sample used was collected closer to shore than



those previously used, all three evaluation procedures show stimulatic



of the algae by taconite alone.








                   EFFECTS OF MANGANESE  ALONE



C-14



     Table 7 and Figure 13 show the  stimulation by manganese.  The



effect is less than ifc, the previous  two  experiments, again probably



because of the proximity of  the sample to shore.






Chlorophyll



     Table 8 and- Figure 13 corroborate the effect of manganese alone.








        EFFECTS OF TACONITE  IN THE PRESENCE OF PHOSPHATE



C-14



     Table 7 shows the combinations  used.  All five  levels of  taconit



 (including 0%) were tested for stimulation against a background  of



both 5 micrograms P/liter and  25 micrograms P/liter.   The  results



are plotted  in Figures 15 and  16  (note scale  differences).



     Figure  15 shows  the taconite effects on  a background  of  5 micro-




grams P/liter.  As expected, just the  addition of P  alone  caused sorm





                            IIQfi

-------
                                20
stimulation over the lake water control.  The addition of .00001%



taconite had no significant effect but concentrations of taconite



of .0001%, .001%, and .01% caused significant stimulation over that



of the phosphorus alone.  These results are replotted in a somewhat



different fashion in Figure 17 showing the results after 20 days of



growth.  Not« that above .00001% the curves for taconite at 0 P and



taconite at 5 micrograms P/liter diverge more rapidly, indicating



that not only i* taconite a more potent stimulator in the presence



of P, but that it becomes proportionately more so at higher concen-



trations of taconite.  Statistical analysis (Keller) shows a highly



significant linear relationship of C-14 uptake to taconite concen-



tration in the presence of SppbP (r=0.80, P=0.017),



     Figures 16 and 18 illustrate the same phenomenon.  Again the



addition of taconite to lake water containing 25 micrograras P/liter



stimulates C-14 uptake and again much more so at higher taconite



concentrations.  Even .00001% taconite in 25 micrograms P/liter



probably is somewhat more stimulatory than 25 micrograms P/liter



alone.  Statistical analysis  (Keller) shows a highly significant



linear correlation  (r=0.97, P=0.009) and highly significant rank



correlation (r=0.90, P=0.0065) of C-14 counts with taconite additions



of 0.00001% to .001%.



     Interestingly,  (Figures  15 and 16) no additions of manganese to



the P were able to provide stimulation as great as that due to addition



of taconite to P.  Once more  this suggests a second growth-promoting



substance in the taconite, other than Mn or P.





Ch1orophy11



     Table 8 summarizes the results as do Figure 19 and 20.  These



data show essentially the same responses as the C-14 uptake.
                            1397

-------
                                21
Algal counts



     The last 2 columns in Table 9 show that, as in Experiment II,



manganese and taconite behave very similarly in a qualitative and



quantitative manner in stimulating algal growth in the presence of




phosphate.  The algae stimulated most were —



     Lyngbya contorta



     Dinobryon sociale



     Stephanodiscus and Cyclotella



     Synedra



     Ceratoneis



     Pi dy mo cy s t is



The totals in the two  flasks were remarkably similar and suggest



that .001% taconite provides at least  4ppb of Mn.






Conclusion



     The  algal growth  stimulation effect of  small  concentrations  of



taconite  is  greatly enhanced by the presence of phosphate  at concen-



trations  of  P  as  low or lower  than  5 micrograms/liter  and  as high or



higher  than  25 micrograms  P/liter.
                            1398

-------
                                22
                    SUMMARY AND CONCUSSIONS




1.  The results of all three experiments establish the fact that



taconite tailings stimulate the growth of the native algae of Lake



Superior when added in concentrations as low as 0.041 milligrams



per liter or as high as 410 milligrams per liter.



2.  The stimulation is evidenced by the increased uptake of C-14,



increased production of chlorophyll, and increased abundance of



the algae at the higher taconite concentrations.



3.  All three criteria show that manganese additions at low concen-



trations stimulate the growth of the algae of Lake Superior.



4.  All three criteria show that phosphate additions at low concen-



trations stimulate the growth of the algae of Lake Superior.



5.  The greater effect of taconite than manganese, the failure of



manganese to increase the effect of taconite, and the great similarities




in response between phosphorus plus manganese, and phosphorus plus



taconite, indicate that one of the active agents of taconite is the



manganese that dissolves from it.



6.  There is evidence for a stimulating agent in taconite that is



neither manganese nor phosphorus.



7.  Addition of phosphorus in concentrations as low as 5 parts per



billion and as high as 25 parts per billion greatly enhances the



stimulatory effects of taconite at all concentrations of taconite



tested.  Stimulation of the growth of algae by taconite in the presence



of phosphorus was as much as 58 fold greater than stimulation by



taconite in the absence of phosphorus.



8.  Most stimulations resulted in significant changes in the relative



abundances of the 4 major groups of algae found in the lake water.
                            1399

-------
                                23
                           REFERENCES




Andrew, R.W., and G.E. Glass, 1970.



     Effect of taconite tailings on algal growth, pp. 73-87 in,



     "Effects of Taconite on Lake Superior", National Water Quality



     Laboratory, Duluth.



Beeton, Ruth, 1973.



     Personal communication.



Glass, G.E., 1970.




     The dissolution of taconite tailings in Lake Superior, pp. 87-



     102 in, "Effects of Taconite on Lake Superior", National Water



     Quality Laboratory, Duluth.



Goldman, C.R., 1966.



     Molybdenum as an essential micronutrient and useful watermass



     marker in Castle Lake, California, pp. 229-238 in, "Chemical



     Environment in the Aquatic Habitat", Koninklijke Nederlandse



     Akademie Van Wetenschappen.



Goldman, C.R., 1970.



     Taconite tailings as a biostimulant for algal growth.  8 pp.



     mimeo.



Keller E., 1973.



     Personal communication.  Data on file in office of J, Shapiro,



Lorenzen,  C.J., 1966.



     A method for  the continuous measurement of  in—vivo chlorophyll



     concentration, Deep Sea Res.  13;223~227.




McGee, Richard, 1970.



     Stimulation of algae growth by  taconite tailings,  23 pp. mimeo,



Schelske,  C,L., L.E.  Feldt, M.A, Santiago, and E, Stoermer, 1972.



     Nutrient enrichment and its effect on phytoplankton production
                            1400

-------
                                24
     and species competition in Lake Superior.  Proc.  15th Conf.



     Great Lakes Res., 1972, pp. 149-



Shapiro, J.,  1970.



     Algae growth studies in Lake Superior, 6 pp. mimeo.



Shapiro, J.,.and G.E, Glass, 1970.



     Chemical factors stimulating growth of Lake Superior algae,



     17 pp.  Paper presented at the 1970 Conference on the Biology



     of Lake Superior.



Vollenweider, R.A., 1969.



     A manual on methods for measuring primary production in



     aquatic environments, International Biological Program, Handbook



     No. 12,  pp. 213.
                           1401

-------
            Table 1.




Carbon-14, net counts per minute
Flask
0%T A
B
C
Dark
.0001%T A
B
C
Dark
.001%T A
B
C
Dark
.01%T A
B
C
Dark
.1%T A
B
C
Dark
2ppbMn A
B
4ppbMn A
B
Dark
SppbMn A
B
16ppbMn A
B
Dark
SppbMn A
*.001%T B
Dark
SppbMn A
+ .1%T B
Dark
1
864
660
1,149
118
881
750
1,113
64
923
849
1,141
94
836
732
1,185
77

744
997
764
816
52
743
954
1,424
1,095
67
1,128
1,017
53

4
2,735
3,088
3,386
148
2,790
2,442
3,609
93
3,351
3,230
3,398
156
3,482
3,363
3,608
112

2,668
3,003
2,976
3,059
78
3,079
2,847
2,897
3,351
117
3,108
3,409
96

Days of Experiment
7 11 14
3,864
4,071
4,088
145
3,690
2,492
4,539
95
5,209
4,192
3,910
107
4,804
5,128
5,720
129

3,910
4,804
3,925
4,071
90
3,470
3,690
4,457
3,925
125
5,024
3,864
200

4,581
4,735
4,973
163
4,283
2,570
5,560
83
6,822
5,623
4,827
100
7,607
7,380
8,594
139

4,758
5,560
5,591
5,321
177
4,690
4,712
4,668
4,718
67
7,446
4,055
109

5,437
5,321
5,328
110
4,948
2,958
6,034
97
8,823
7,219
6,966
80
10,499
9,973
10,611
284

5,154
7,786
7,492
6,822
174
5,236
6,383
5,890
6,966
77
6,136
9,497
109

19
7,167
7,065
7,219
146
7,065
5,787
8,982
185
14,258
11,872
8,038
162
15,357
16,366
16,640
135
8,170
6,302
7,606
105
5,823
11,878
12,319
11,337
105
7,910
9,407
8,306
9,688
132
10,074
13,672
148
8,520
6,730
112
            1402

-------
  Table 2.



Chlorophyll
Days of Experiment
Flask
0%T A
B
C
Dark
.0001%T A
B
C
Dark
.001%T A
B
C
Dark
.01%T A
B
C
Dark
.1%T A
B
C
Dark
2ppbMn A
B
4ppbMn A
B
Dark
SppbMn A
B
16ppbMn A
B
Dark
SppbMn A
+.001%T B
Dark
SppbMn A
+ ,1%T B
Dark
0
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
19.5
19.5
19.5
19.5
25.0
25.0
25.0
25.0
39.0
39.0
39.0
39.0
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
18.5
39.0
39.0
39.0
4
15.0
12.0
11.9
20.5
13.0
9.0
14.0
15.0
25.0
20.0
10.5
20.0
23.0
22.5
20.0
27.0
35.4
39.0
35.0
40.0
14.0
16.0
13.0
15.0
21.0
11.5
9.0
25.4
14.5
12.5
13.0
15.0
21.5
37.5
39.5
27.0
7
13.5
18.0
19.5
13.0
13.0
13.0
20.0
19.0
23.0
16.0
19.0
13.0
23.5
22.5
23.0
16.0
38.5
40.5
41.0
33.0
17.0
19.0
15.5
16.0
21.0
16.0
13.0
16.0
11.0
10.5
12.5
27.0
12.5
43.0
45,0
32.5
11
15.0
21.0
15.0
17.0
13.5
8.0
16.0
10.5
24.5
22.0
14.0
9.5
35.0
34.5
32.0
14.0
47.0
43.5
46.0
31.0
14.0
21,0
23.0
26.0
14.5
15,5
20.0
18.0
21.0
7.5
21.5
27.0
18.5
48.0
47,0
30.5
14
21.0
19.5
20.5
9.0
19.5
21.5
24.0
8.0
37.0
23.5
20.5
8.0
46.0
34.0
32.0
13.0
52.0
46.0
53.0
23.0
21.0
31,0
27.5
26.5
11,0
20.0
23,5
24,0
20.0
7.0
24,0
30,0
14,0
51.5
50.0
30.0
19
19.0
25.0
9.0
26.0
26.5
29.0
8.5
43.0
37.0
31.5
11.0
50.5
48.0
16.0
63.5
58.0
72.0
29.0
27.0
40,5
46,0
35.0
8,0
26.0
31.5
31.5
35.0
7.0
28.0
43,0
10.0
61.5
62,0
30.0
 1403

-------
                            Table 3.
                          Algae Counts
                   Lake Superior Experiments

                    Aug 18 - Sept 6, 1972
Alga

   (Chrysophyta)
Fragilaria crotonensis
Asterionella formosa
Nitzschia actinastroides
Rhizosolenia eriensis
R. longiseta
small Cyclotella & Stephanodiscus
Dinobryon bavaricum
D. socials
D. divergens
Synedra spp.
small chrysophyte flagellates
                         Total
    (Cyanophyta)
Aphanothece clathrata
Synechococcus elongatus
Oscillatoria sp.
                         Total
    (Chlorophyta)
Oocystis spp.
Elakatotrix gelatinosa
Didymocystis sp.
Ankistrodesmus  spirilliformis
A.  falcatus
Tetraedron minimum
Chlamydomonas sp.
                          Total

    (Cryptomonads  &  y  flagellates)
Cryptomonads 1
             2
             3
yflagellates

                          Total
                                                Nurrojers/ml
                                                  Sample
Original
day 0
4


16
4
116
15
20
15

17
207
2
60

62
7
4
28

2
4
13
58
i
49
71
36
80 '
236
Control A,
day 19
143
30
23
225

307
68
30
53

82
961
7
67
11
85


30
7
4
4
7
52
90
30

86
206
.001% tai
day
270
90

108

1387
38
251
41
4
45
2234


4
4


68

19


~8T
64
45

' 83
192
                    GRAND TOTAL
563
1304
2517
                            1404

-------
            Table 4.



Carbon-14, net counts per minute
Flask
0%T A
B
C
Dark
.0001%T A
B
C
Dark
.001%T A
B
C
Dark
,01%T A
B
C
Dark
lOppbP A
B
Dark
20ppbP A
B
Dark
20ppbP A
+.001%T B
Dark
20ppbP A
+ ,01%T B
Dark
SppbMn A
B
Dark
SppbMn A
+.001%T B
Dark
SppbMn A
+ 20ppbP B
2
1,827
1,782
2,256
45
1,791
2,119
1,828
60
2,152
2,048
2,171
58
1,819
1,949
1,898
62
2,511
2,348
41
1,905
2,577
49
2,166
2,424
68
1,639
1,889
45
1,908
2,026
29
2,124
2,299
39
2,388
2,454
Da1
.,_ ... ji
5
3,401
3,621
4,437
48
4,046
4,005
4,030
66
3,727
3,880
4,603
61
3,792
3,919
4,646
85
6,165
6,071
70
4,247
5,943
72
5,687
7,246
88
5,024
5,671
95
3,969
4,148
64
4,508
4,408
273
5,979
6,085
/s of Experiment
10
5,671
5,704
6,488
44
6,507
6,363
6,662
79
6,383
6,917
7,520
69
6,467
7,015
7,973
70
14,788
18,322
45
9,497
11,601
118
25,289
29,385
83
18,429
23,502
81
6,243
6,252
56
7,191
7,141
52
16,233
21,250
16
7,120
7,743
9,275
78
8,054
7,687
9,275
85
8,862
9,254
9,705
102
10,390
10,390
12,434
105
23,783
35,687
157
20,806
24,664
131
56,311
68,939
225
65,548
83,306
163
8,153
9,105
80
9,407
9,382
103
43,457
54,768
21
9,064
9,029
8,789
83
9,085
8,484
10,472
100
9,168
10,872
11,402
107
13,814
15,067
12,960
113
30,742
48,166
96
38,068
48,753
55
90,909
102,564
103
105,263
111,111
183
9,520
11,534
70
10,444
10,811
137
68,966
81,633
            1405

-------
  Table  5,




Chlorophyll
Flask
0%T A
B
C
Dark
.0001%T A
B
C
Dark
.001%T A
B
C
Dark
.01%T A
B
C
Dark
lOppbP A
B
Dark
20ppbP A
B
Dark
20ppbP A
+.001%T B
C
20ppbP A
+ ,01%T B
Dark
SppbMn A
B
Dark
SppbMn A
+.001%T B
Dark
SppbMn A
+ 20ppbP B
2
16.0
18.0
17.0
15.0
14.0
15.5
15.0
13.0
17.0
17.0
18.0
15.0
22.0
21.0
22.5
19.0
17.0
18.5
13.0
16.0
18.0
13.5
17.5
19.0
13.5
22.0
22.0
19.0
16.0
15.0
13.0
16.5
18.5
14.0
20.0
18.5
Da;
5
20.0
19.0
18.0
11.5
21.0
25.0
21.0
11.5
21.0
21.0
22.5
12.5
23.5
26.0
27.0
17.0
35.0
33.5
12.0
32.0
38.5
12.5
31.0
42.0
12.0
32.0
39.0
16.5
20.0
21.0
12.0
20.0
20.5
11.5
37.5
31.0
£S_ of Experiment
10 16
23.5
26.5
28.0
8.0
26.0
26.0
26.0
12.0
25.5
26.0
25.5
7.5
33.5
36.0
37.0
12.0
77.0
95.0
8.0
64.5
80.5
7.5
8.0
12.5
27.5
27.5
8.0
30.5
31.5
8.5
90,0
94.5
21.5
23cO
27.0
8.5
28.0
28.0
26.0
9.0
31.5
30.0
25.0
11.0
39.5
33.5
42.5
14.0
140
160
6.0
148
170
6.0
352
316
7.5
376
384
12.0
24.0
23.0
9.0
29.5
28.0
9.5
258
243
21
24.0
26.0
26.0
7.5
28.0
26.0
25.0
7,0
28.0
28.0
28.0
7.5
43.0
39.0
41.5
14.0
175
220
6.5
312
370
6.0
410
370
7.0
425
346
11.5
26.5
25.0
6.0
27.0
24.0
8.5
385
385
  1406

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3-
                                                      1408

-------
            Table 7.
Carbon-14, net counts per minute
Flask
0%T A
B
.00001ST A
.0001%T A
B
,001%T A
B
.01%T A
B
Dark
2ppbMn A
B
4ppbMn A
B
SppbMn A
B
Dark
SppbP A
B
SppbP A
+.00001%T B
SppbP A
+.0001%T B
5ppbP+.001%T A
5ppbP+.01%T A
2 SppbP A
B
2 SppbP A
+.00001%T B
2 SppbP A
+.0001%T B
25ppbP+.001%T A
2 SppbP A
+ . 01%T Dark
5ppbP+4ppbMn A
5ppbP+ SppbMn A
25ppbP+4ppbMn A
2 5ppbP+ SppbMn A
2 SppbP A
+4ppbMn B
1
275
273
220
199
319
218
274
218
176
5
235
254
291
296
229
270
7
210
232
285
231
303
237
360
279
198
225
239
251
212
274
212
240
11
181
269
205
221
303
271
Day
5
1,385
1,242
1,435
1,410
1,337
1,481
1,455
1,506
1,443
19
1,490
1,376
1,337
1,378
1,533
1,261
11
1,819
1,641
1,633
1,684
1,608
1,343
1,773
1,831
1,729
1,689
1,720
1,779
1,742
1,639
1,467
1,663
29
1,989
1,580
2,014
1,698
1,815
1,558
rs of experiment
11 15
2,160
1,963
2,549
2,636
2,076
2,678
2,664
2,761
2,840
75
2,639
2,554
2,102
2,210
2,784
1,964
58
5,013
4,684
3,983
4,762
3,902
4,435
5,195
5,319
4,329
5,277
5,025
5,464
6,135
4,098
6,369
5,495
54
6,135
3,614
6,472
5,155
5,682
4,329
2,528
2,503
3,155
2,837
2,496
3,152
3,027
3,795
3,849
57
2,722
3,019
2,555
2,603
3,003
2,169
40
5,720
5,821
5,487
6,108
6,684
6,574
7,973
8,541
5,423
6,893
6,445
7,246
10,229
7,607
15,358 .
15,175
39
7,786
3,616
11,669
9,777
15,011
10,902
20
2,861
3,330
3,365
3,384
2,577
3,671
3,517
4,804
4,746
37
3,289
3,168
2,783
3>019
3,296
3,219
36
7,167
7,090
6,302
6,684
9,064
6,730
9,777
11,534
6,022
10,023
8,412
10,282
16,780
15,598
37,037
35,061
77
8,745
7,167
25,947
24,363
32,760
25,947
            1409

-------
  Table 8.
Chlorophyll
Flask
0%T A
B
.00001%T A
.0001%T A
B
,001%T A
B
.01%T A
B
Dark
2ppbMn A
B
4ppbMn A
B
SppbMn A
B
Dark
Sp'pbP A
B
SppbP A
+.00001%T B
SppbP A
+ .0001%T B
5ppbP+.001%T A
5ppbP+.01%T A
2 SppbP A
B
2 SppbP A
+.00001%T B
2 SppbP A
+.0001%T B
25ppbP+.001%T A
25ppbP+.01%T A
Dark
5ppbP+4ppbMn A
5ppbP+8ppbMn A
25ppbP+4ppbMn A
2 5p_pbP+ SppbMn A
2 SppbP
+4ppbMn A
+.01%T B
1
14.0
14.5
14.5
14.5
14.5
14.5
16.5
21.5
20.0
20.5
14.0
14.5
14.5
14.0
15.0
14.5
14.5
14.5
16.5
14.5
14.0
15.0
15.0
15.5
20.5
13.5
14.5
14.5
14.0
16.0
15.0
15.5
21.0
20.5
13.0
14.0
14.0
15.0
21.0
20.5
Days of experiment
5 11 15
18.5
16.5
19.5
28.0
20.0
21.0
20.0
27.0
25.0
17.0
19.5
23.0
18.0
18.0
22.0
18.0
12.0
33.0
32.0
29.5
27.0
23.0
24.5
28.5
34.5
27.5
29.0
34.0
30.0
37.0
26.5
26.0
36.0
16.0
30.0
25.0
37.0
30.0
39.0
32.5
26.0
2a.s
30.0
28.0
25.5
28.0
25.0
35.0
33.0
15.5
29.0
28.5
24.0
24.0
30.0
23.0
10.5
69.0
72.0
68.0
77.0
78.0
81.5
83.0
96.0
75.0
93.0
88.0
95.0
126
88.0
99.5
101.0
15.0
85.5
61,0
122
90.0
116
112
31.5
26.0
29.5
28.0
27.0
31.5
31.0
42.0
40.0
13.0
25.5
29.0
28.0
26.0
29.0
23.5
7.0
55.0
63.5
84.0
80.5
76.0
76,5
86.0
115
68.5
84.5
83.5
98.5
168
185
327
345
12.5
69.0
66.5
155
160
260
250
20
26.5
24.5
27.0
24.5
28.5
33.0
27.0
48.0
40.0
12.5
24.0
30.0
23.5
25.5
28.0
23.0
6.5
65.0
67.0
73.0
69.0
91.5
72.0
81.0
130
165
195
180
180
245
335
375
620
12.0
69.0
66,0
315
345
428
468
  1410

-------
                            Table 9.
                          Algae Counts
                   Lake Superior Experiments
                      Nov 2 - Nov 22, 1972
                                           Numbers/ml
                                             Sample
   (Chrysophyta)
Fragilaria spp.
Asterionella formosa
Rhizosolenia eriensis
Tabellaria flocculosa
Synedra ulna.
Synedra spp.
Stephanodiscus-Cyclotella
Stephanodiscus niagarae
Nitzschia spp.
Ceratoneis sp.
Navicula spp.
Dinobryon sociale
Dinobryon bavaricum
Dinobryon divergens

               Total

   (Cyanophyta)
Oscillatoria planctonica
Lyngbya contorta
Lyngbya limnetica
Microcystis aeruginosa
Microcystis incerta

               Total

   (Chlorophyta)
Didymocystis sp.
Ankistrodesmus falcatus
A. falcatus var. spi:
               Total

   (Cryptomonads & f:
Cryptomonads 1
             2
Euglena sp.

               Total
         GRAND TOTAL
Original
day 0

6


13
15
la 13


2
2


2
53
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0
49

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4
6
2
12
114
Control A
day 20
17

9

24
71
11
2

4
4
118
7
	 43
310

67
4
17

88
722
2

724
9
43
	 2
54
1176
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day 20
24
79
13

101
52
58

2
7
4
71
4
	 52_
467
6
43

7
	 13
69
1320
4

1324
4
41

45
1905
,001%T+25P
day 20

210

45
3300
5400
91J950


255

1200


102,360

315

105
	 15
435
10,050

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10,080

75

75
112,950
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day 20

105


450
1200
91,350


285

135


93,525

450
15


465
16,050
60
	 T5
16,125

210

210
110,325
                           1411

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