r
BIOLOGICAL PROBLEMS
IN WATER POLLUTION
U.S. DEPARTMENT OE
HEALTH, EDUCATION, AND WELFARE
Public Health Service
1957
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BIOLOGICAL PROBLEM S
I N
WATER POLLUTION
Transactions of a Seminar on Biological Problems
in Water Pollution held at the
Robert A. Taft Sanitary Engineering Center
C i nc i nnat i, Oh i o
April 23 - 27, 1956
Compiled and Edited
by
CLARENCE M. TARZWELL
US EPA
Headquarters and Chemical Libraries
EPA West Bldg Room 3340
Mailcode 3404T
1301 Constitution Ave NW
Washington DC 20004
U.S. 0«»
HEALTH, EDUCATION, AND WELFARE
Public Health Service
Bureau of State Services
Division of Sanitary Engineering Services
Robert A. Taft Sanitary Engineering Center
I 957
Repository Material
Permanent Collection
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PREFACE
During the past few years a number of State Health Departments and
Water Pollution Control Boards have initiated or expanded investigations in
the field of sanitary biology. Universities and other research organizations
are showing increased interest in biological problems connected with the
detection and abatement of stream pollution. A few universities are now
giving courses directed toward the training of sanitary biologists, and
several are considering the establishment of curricula for the training of
aquatic biologists for work in the water works, sewage treatment, and
stream pollution fields. In recent years industries have added sanitary
biologists to their staffs, and several aquatic biologists have undertaken
consultant activities.
Biologists engaged in pollution investigations and research often work
alone and are somewhat isolated. For some time, therefore, there had been
recognized a need for a conference of those engaged in the study of biological
problems in water pollution control, to acquaint them with current developments
and new methods of approach, and to enable them to become acquainted with other
workers in the field. The first such gathering was held as a seminar at the
Robert A. Taft Sanitary Engineering Center, April 23 - 27, 1956. The meeting
was well attended, as ninety persons were registered at the Seminar. Repre-
sentatives were present from twenty-eight states, the District of Columbia,
and the Provinces of Ontario, Quebec, and New Brunswick, Canada. The
following states were represented: Alabama, California, Florida, Georgia,
Idaho, Illinois, Indiana, Iowa, Louisiana, Massachusetts, Michigan, Minnesota,
Mississippi, Missouri, Montana, New York, North Carolina, Ohio, Oregon,
Pennsylvania, Rhode Island, South Carolina, Tennessee, Texas, Utah, Washing-
ton, Wisconsin, and Wyoming. Participants included workers from industry,
State Conservation Departments, Universities, State Boards of Health, and
Stream Control Commissions. Representatives were also present from the
Ohio River Valley Water Sanitation Commission, the Illinois State Natural
History Survey, the Atomic Energy Commission, the U.S. Naval Radiological
Defense Laboratory, &nd the Fish and Wildlife Service. A total of twenty-
three were in attendance from the Public Health Service, six from outside
the Cincinnati Area.
The Dow Chemical Company, Atlantic Refining Company, Sulphite Pulp
Mfg. Research League, Institute of Paper Chemistry, General Electric
Company, and Pantech, Inc. had representatives at the Seminar. Representatives
were present from Conservation Departments of the following states: Alabama,
Idaho, Indiana, Iowa, Michigan, Mississippi, Missouri, Montana, New York,
Ohio, Pennsylvania, Tennessee, and Wyoming.
Representatives were present from sixteen universities as follows:
Pennsylvania State University, University of Florida, Massachussets Institute
of Technology, University of Missouri, University of Wisconsin, Michigan
State University, University of Utah, Ohio State University, University of
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Michigan, University of Miami, University of Southern California, University
of Montreal, University of Minnesota, University of Cincinnati, University
of Toronto, and University of North Carolina. The State Boards of Health
of Florida, Missouri, and ILlinois sent representatives as did the Stream
Control Commissions of Louisiana and Wisconsin. Michigan led in the
number of representatives present, having seven; Ohio was second with
six; the Province of Ontario, Pensylvania, and California were third with
five each; Wisconsin, Florid^., and Alabama each had four; while Iowa
and Missouri sent three.
The seminar consisted of panel discussions and was planned so that
most of the time was devoted to commentary from the floor with only short
presentations by panel members. Subjects discussed were: (1) Use and
Value of Bioassays; (2) Use and Value of Biological Indicators of Pollution;
(3) Current Investigations in Water Pollution Biology; (4) Water Quality
Criteria for Aquatic Life; •'and (5) Training of Sanitary Aquatic Biologists.
Discussions of each subject were lead by a panel chairman and 4 to 9 panel
members, each of whom presented a phase of the problem in 10 to 20 minutes .
The remainder of the day was then devoted to discussions from the floor. All
presentations and discussions were off the record in order that the participants
could feel free to express their opinions. All members of the panel on
Biological Indicators of Pollution were asked to prepare papers to be included
in the transactions of the meeting. These papers could be discussed in their
panel presentations if they wished or their presentations could be entirely
different. All panel metnbers and all those in attendance were invited to
submit papers for inclusion in the transactions. The transactions, therefore,
are not a record of the discussions at the meeting, and the papers contained
therein may or may not have been presented during the panel discussions,
but are the results of the prospect or the events of the sessions.
During the meeting, discussions were free and critical. It is believed
they were valuable in stimulating those in attendance and will serve to advance
research and investigations in the field of water pollution. Numerous letters
have been received expressing approval of the meeting and inquiring as to
when another similar meeting would be held. It is planned to hold a seminar
on biological problems in water pollution every three years; hence, the next
meeting will occur in 1959. It is requested that all those interested in such
a meeting submit their suggestions by June 1958. Suggestions are solicited
as to time and length of meeting, program, and manner of carrying on the
discussions.
C. M. Tarzw^ll
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TABLE OF CONTENTS
Page
Preface. 1
List of those in attendance 6
Picture of group 17
USE AND VALUE OF BIO-ASSAYS 18
Derivation of the threshold value for toxicity and the equation
of the curve by a graphical method - George Burdick 19
Interim plan for standardizing the bioassay of paralytic shellfish
poison by use of a reference standard - Public Health Service 22
Application factors to be applied to bioassays for the safe
disposal of toxic wastes - Croswell Henderson 31
USE AND VALUE OF BIOLOGICAL INDICATORS OF POLLUTION 38
Some problems in the identification of microorganisms -
Wm. Bridge Cooke 39
Use and value of bacterial indicators of pollution - Paul Kabler 43
Protozoa as indicators of the ecological condition of a body
of water - James B. Lackey 50
Algae as biological indicators of pollution - C. Mervin Palmer 60
Diatoms as indicators of changes in environmental
conditions - Ruth Patrick 71
Use and value of fungi as biological indicators of
pollution - Wm. Bridge Cooke 84
Use and value of biological indicators of pollution: Freshwater
clams and snails - William Marcus Ingram 94
The use and value of aquatic insects as indicators of
organic enrichment - Arden R. Gaufin 136
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Page
Biological indices of water pollution, with Special reference to
fish populations - Peter Doudoroff and Charles E. Warren 144
Biological criteria for the determination of lake pollution -
Eugene W. Surber 164
The use and abuse of indicator organisms - William M. Beck, Jr 175
CURRENT INVESTIGATIONS IN WATER POLLUTION BIOLOGY 178
Current water pollution investigations and problems in
Wisconsin - Kenneth M. Mackenthun. 179
Investigations and problems in Ontario - John H. Neil 184
J&eport on pollution studies conducted in Western
Canada - Michael Waldichuk 188
The Telationship of the polychaetous Annelid Capitella
capitata (Fabricius) to waste discharges of biological
origjiv *¦ Donald J. Reish 195
Cooperative research at Oregon State College in the
biological aspects of water pollution - Charles E. Warren and
Peter Doudoroff 201
Some aspects of water pollution in the Missouri Basin -
Joe K. Neel * 209
Investigations and Problems in Ohio - John N. Reis 215
Biology and water pollution in Great Britain - Thomas Beak 220
THE TRAINING OF AQUATIC SANITARY BIOLOGISTS 224
The training of aquatic sanitary biologists -
Curtis L. Newcombe 225
The training of aquatic sanitary biologists - a letter -
Charles E. Renn 230
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Page
WATER QUALITY CRITERIA FOR AQUATIC LIFE 234
Effects of turbidity and silt on aquatic life -
John N. Wilson 235
The effect of pollution upon wildlife - O. Lloyd Meehean 240
Water quality criteria for aquatic life - Clarence M. Tarzwell 246
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PR_0_GR A_M
SEMINAR ON BIOLOGICAL PROBLEMS IN WATER POLLUTION
April 23 - 27, 1956
DATE & TIME SUBJECT
Monday
April 23
8:00 - 9:00 REGISTRATION
9:00 - 9:15 WELCOME - Harry G. Hanson, Director
Robert A. Taft Sanitary Engineering Center
9:15 - 9:30 PLAN OF THE SEMINAR AND ANNOUNCEMENTS -
Clarence M„ Tarzwell
Chief of Aquatic Biology
9:30 - 4:30 Panel Discussion - Use and Value of Bio-Assays
Chairman, George Burdick, Dept. of Conservation
Albany, New York
Extending Acute Toxicity Data to Indicate Toxicity Under
Continuous Exposure
George Burdick
Department of Conservation
Albany, New York
Bio-Assays for Determining the Tainting of Fish Flesh
A. W. Winston
Dow Chemical Company
Midland, Michigan
Use of Bio-Assays in Industry
W. B. Hart
Director, Industrial Waste Eng. Div.
Pantech, Inc.
Folcroft, Pennsylvania
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DATE & TIME
SUBJECT
Monday-
April 23 (cont'd) Methods of Assav for Paralytic Shellfish Poison
Earl F, McFarren
Public Health Service
Cincinnati, Ohio
Application Factors to be Applied to the Bio-Assays for
the Safe Disposal of Toxic "Wastes
Croswell Henderson
Public Health Service
Cincinnati Ohio
8:00 P, M, Social Evening at Residence of C. M. Tarzwell
780 Ivy Ave.
Glen dale., Ohio
Tuesday
April 24
8:30 - 4:30 Panel Discussion - Use and Value of Biological
Indicators of Pollution
Chairman. Dr. A. R, Gaufin; University of Utah,
Salt Lake City. Utah
Problems in Identification of Microorganisms
Dr. Wm Bridge Cooke
Public Health Service
Cincinnati Ohio
Bacteria
Dr. Paul Kabler
Public Health Service
Cincinnati, Ohio
Protozoa
Dr, James Lackey
University of Florida
Gainesville, Florida
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DATE & TIME
SUBJECT
Tuesday
April 24 (cont'd) Algae
Dr C Mervin Palmer
Public Health Service
Cincinnati, Ohio
Fungi
Dr. Wm. Bridge Cooke
Public Health Service
Cincinnati;, Ohio
Mollusca
Dr. "William Ingram
Public Health Service
Cincinnati, Ohio
Macro-Invertebrates
Dr. A„R. Gaufin
University of Utah
Salt Lake City, Utah
Fishes
Dr. Peter Doudoroff
Public Health Service
Oregon State College
Corvallis, Oregon
Indicators of Lake Pollution
Eugene Surber
Fish and Wildlife Service
Atlanta, Georgia
Wednesday
April 25
8:30 - 4:30 Panel Discussion - Water Quality Criteria for Aquatic
Life
Chairman, Dr. O. Lloyd Meehean
Assistant to the Director, U.S. Fish and Wildlife
Service
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DATE & TIME
SUBJECT
Wednesday
April 25 (cont'd)
Oxygen Requirements
Dr. William Spoor
University of Cincinnati
Cincinnati, Ohio
Effects of Turbidity and Silt
John Wilson
Public Health Service
Portland, Oregon
Requirements for Aquatic Life
Dr. O. Lloyd Meehean
Fish and Wildlife Service
Washington, D. C.
The Status of Water Quality Criteria
Dr. C. M. Tarzwell
Public Health Service
Cincinnati, Ohio
6:00
Group Supper - Millcroft Inn, Milford, Ohio
Thursday
April 26
8:30 - 4:30 Panel Discussion - Current Investigations in Water
Pollution Biology
Chairman, Kenneth M. Mackenthun, Committee on
Water Pollution, Madison, Wisconsin
Investigations and Problems in the Missouri Basin
Dr. J.K. Neel
Public Health Service
Kansas City, Missouri
Investigations and Problems in Alabama
Dr. I. B. Byrd
Alabama Dept. of Conservation.
Montgomery, Alabama
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DATE & TIME
SUBJECT
Thursday
April 26 (cont'd) Biology and Water Pollution in Great Britain
Thomas W„ Beak
Consulting Biologist
Hawkesbury, Ontario
Investigations at Oregon State College
Charles "Warren
Department of Fish and Game Management
Oregon State College
Corvallis, Oregon
Investigations and Problems in Ontario
John Neil
Ontario Dept. of Health
Toronto, Ontario, Canada
4:30 - 6:00 Program Review and Tour of the Robert A. Taft Sanitary
Engineering Center
6:30 Dinner - Center Cafeteria
Speaker - Harry A. Faber, PHS, Washington, D. C.
The PHS Research Grant Program
8:00 P.M. Panel Discussion - The Training of Aquatic Sanitary
Biologists
Chairman, Dr. Herbert Jackson, Public Health
Service, Cincinnati, Ohio
Panel Members:
Dr. Curtis L. Newcombe
U. S. Naval Radiological Defense Laboratory
San Francisco, California
Dr. Lloyd Smith
University of Minnesota
St. Paul, Minnesota
Dr. Charles Renn
Department of Sanitary Engineering and Water Resources
Johns Hopkins University
Baltimore, Maryland
Dr. T.H. Langlois
Ohio State University
Columbus, Ohio
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DATE & TIME
SUBJECT
Thursday-
April 26 (cont'd)
Dr William Spoor
University of Cincinnati
Cincinnati Ohio
Friday
April 27
9:00 - 4:30 Panel Discussion - Current Investigations in Water
Pollution Biology
Chairman, Dr. A. F. Bartsch, Public Health
Service, Cincinnati, Ohio
Use and Value of Sewage Lagoons
Dr , A , F, Bartsch
Public Health Service
Cincinnati, Ohio
Investigations and Problems in Florida
Mr William M, Beck
State Board of Health
Jacksonville. Florida
Investigations and Problems in Illinois
Dr William C Starrett
Illinois Natural History Survey
Urbana, Illinois
Investigations and Problems in the Disposal
of Radioactive Wastes
Dr, J.J, Davis
General Electric Company
Richland., Washington
Investigations and Problems in Ohio
John Ries
Division of Wildlife
Dept of Natural Resources
Columbus, Ohio
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DATE & TIME
SUBJECT
F riday
April 27 (cont'd)
The Relationship of the Polychaetous Annelid
Capitella capitata (Fabricius) to Waste
Discharges of Biological Origin
Donald J. Reish
Dept. of Biology & the Allan Hancock Foundation
University of Southern California
Los Angeles, California
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List of Registrants
Seminar on Biological Problems in Water Pollution
April 23-27 1956
Public Health Service, R A Taft Sanitary Eng. Center
Cincinnati, Ohio
Dr. Bertil G. Anderson
Pennsyl:^ania State University
University Park,
Pennsylvania
J, B, Anderson
PHS, Region 7
Dallas, Texas
Robert F, BaJch
Inst, of Paper Chemistry
Appleton, Wis.
Dr, Robert C, Ball
Div, of Conservation
Michigan State College
E, Lansing, Mich.
P.G. Barnickol
Missouri Conservation Comm
Columbia, Mo.
Dr. A, F. Bartsch
PHS
Cincinnati, Ohio
T . W. Beak
4 Hamilton Street
Hawkesbury, Ont.
William M Beck Jr
Bureau of San Eng.
Florida State Bd. of Health
Jacksonville, Fla.
B. B. Berger
PHS
Cincinnati, Ohio
L, Berner
University of Florida
Gainesville, Fla.
K„ E, Biglane
Stream Control Comm.
Baton Rouge, La.
Mario Boschetti
Lawrence Experiment Station
Lawrence, Mass.
Clifford E, Bosley
Game and Fish Commission
Cheyenne, Wyo.
Arthur Bradford
Pennsylvania Fish Comm.
Harrisburg, Pa.
G.E, Burdick
Conservation Department
Albany 1, N. Y.
I . B. Byrd
Dept of Conservation
Montgomery, Ala.
Dr. Robert S Campbell
University of Missouri
Columbia, Mo.
Robert Cleary
Biology Bldg,
Okoboji, Iowa
Dr. Wm, Bridge Cooke
PHS
Cincinnati, Ohio
J. Davi s
General Electric Co,
Richland, Wash.
Dr, Peter Doudoroff
PHS
Corvallis, Ore.
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Robert J. Ellis
Harry Faber
Dr Frederick Fish
Dr George P Fitzgerald
Dr Paul O Fromm
Dr . Arden R, Gaufin
Dick Graham
G, Hamlin
Harry Hanson
Harry Harrison
W, B Hart
Forrest Hauck
Croswell Henderson
Karl E0 Herde
John Hester
Dr. Frank F„ Hooper
W. Charles Howard
Dr, Wm, Ingram
Dr, Herbert Jackson
Dr. Paul Kabler
H D. Kelly
Dr, M, H.A, Keenleyside
Harry Kramer
Dr, James B, Lackey
Dr. Thomas H. Langlois
Inst. for Fisheries Research
Michigan Dept. of Conservation
Eng. Resources Program. PHS
PHS
University of "Wisconsin
Michigan State University
University of Utah
Dept. of Fish and Game
ORSANCO
PHS
Biology Building
Industria] Waste Eng , Div
Pantech, Inc
Dept of Fish and Game
PHS
AEC
Dept. of Conservation
Michigan Dept. of Conservation
PHS
PHS
PHS
PHS
Dept. of Conservation
Fish. Research Bd. of Canada
PHS
University of Florida
Ohio State University
Ann Arbor, Mich.
"Washington D. C,
Atlanta, Ga.
Madison, "Wis.
E, Lansing. Mich.
Salt Lake City, Utah
Helena, Montana
Cincinnati Ohio
Cincinnati, Ohio
Okoboji, Iowa
Folcroftj Pa.
Boise, Idaho
Cincinnatij Ohio
Aikenj S. Carolina
Montgomery, Ala.
Ann Arbor, Mich.
Cincinnati, Ohio
Cincinnati, Ohio
Cincinnati, Ohio
Cincinnati, Ohio
Montgomery. Ala.
St. Andrews, N. B„
Cincinnati, Ohio
Gainesville, Fla.
Columbus, Ohio
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Dr. George H. Lauff
University of Michigan
Ann Arbor, Mich.
Bernard Lueck
Sulphite Pulp Mfg. Res. League
Appleton, Wis.
Kenneth M. Mackenthun
Comm. on Water Pollution
Madison.. Wis.
Earl McFarren
PHS
Cincinnati, Ohio
Kneeland McNulty
Marine Laboratory
Coral Gables, Fla .
H„E. McReynolds
Ind. Dept. of Conservation
Versailles, Ind,
Dr. O, Lloyd Meehean
Fish & Wildlife Service
Washington, D. C.
R.H. Millest
Dept. of Lands and Forests
Toronto, Ontario
John L, Mohr
Univ, of Southern California
Los Angeles, California
D, D, Moss
Dept of Conservation
Montgomery, Ala.
J„ K Nee]
PHS, Region 6
Kansas City, Mo.
John H. Neil
Ontario Dept. of Health
Toronto, Ontario
Dr, Curtis L„ Newcombe
U S Naval Radiological Def. Lab,
. San Francisco, Calif.
Dr, H, P. Nicholson
PHS, Region 6
Kansas City, Mo.
George Paine
PHS
Cincinnati, Ohio
Ralph Palange
PHS
Cincinnati, Ohio
Dr. C. Mervin Palmer
PHS
Cincinnati, Ohio
Quentin Pickering
PHS
Cincinnati, Ohio
Gustave Prevost
University of Montreal
Montreal, Canada
R. Raneri
PHS
Cincinnati, Ohio
John N, Reis
Aquatic Biology Laboratory
Delaware, Ohio
Dr. S. C. Rittenberg
University of Southern Calif.
Los Angeles, Calif.
Earl T. Rose
Biology Building
Okoboji, Iowa
Donald J. Reish
University of Southern Calif.
Los Angeles, Calif.
J. D. Roseborough
Lands and Forests
Toronto, Ontario
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E, R. Roth
C. E. Ruhr
Robert Schiffman
Lloyd L. Smith, Jr.
Wesley E. Smith
William G. Spence
Dr, Wm. Spoor
John Sprague
Dr. William C. Starrett
Dr. Robert E. Stevenson
Harold Streeter
Eugene W. Surber
Dr. C. M. Tarzwell
W. W. Towne
William J. Tucker
John E. Watson
Charles M. Weiss
John N. Wilson
A. W. Winston, Jr.
Charles B. Wurtz
Atlantic Refining Co.
State Game & Fish Comm.
Michigan State University
University of Minnesota
Bur. of Public Health Eng.
Division of Health
Game and Fish Commission
University of Cincinnati
University of Toronto
State Nat. Hist. Survey Div.
University of Sourthern Calif.
ORSANCO
Fish and Wildlife Service
PHS
PHS
Dept. of Public Health
Fish and Wildlife Service
University of North Carolina
PHS
Dow Chemical Company
Commercial Trust Building
Philadelphia, Pa.
Nashville, Tenn.
E. Lansing, Mich.
St. Paul, Minn.
Jefferson City, Mo.
Jackson, Miss.
Cincinnati, Ohio
Toronto, Ontario
Urbana, Illinois
Los Angeles, Calif.
Cincinnati, Ohio
Atlanta, Georgia
Cincinnati, Ohio
Cincinnati, Ohio
Springfield, 111.
E. Greenwich, R.I.
Chapel Hill, N. C.
Portland, Oregon
Midland, Michigan
Philadelphia, Pa.
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1.
2.
3.
U.
5-
6.
7.
8.
9.
10.
11.
12.
13-
llj.
15.
16.
17.
IS.
iy.
John Wilson
John N. Reis
S. C. Rittenberger
Robort E. Stevenson
Donald J. Reish
Quentin Pickering
John Hester
H. D. Kelly
I. B. Byrd
D. D. Moss
George H. Lauff
T. W. Beak
Croswell Henderson
Mario Boschetti
J. B. Anderson
Clifford E. Bosley
William M. 3eak, Jr.
Bernard Lueck
A. F. Dartsch
20. J. D. Roseborough
21. Charles M. Weiss
22. John H. Neil
23. William C. Starrett
2h. John Sprague
25. James B. Lackey
26. Karl E. Herde
27. H. E. McReynolds
28. Arthur Bradford
29. J. J. Davis
30. A. W. Winston
31• Kneeland McNulty
32. George E. Burdick
33. M. H. A. Keenleyside
3U« Kenneth M. Macksnthun
35.
36.
37.
38.
39.
ao.
bl.
I4 2.
U3.
Arden R. Gaufin
Harry Faber
Georgo P. Fitzgerald
Lloyd L. Smith, Jr.
R. H. Millest
Bertil G. Anderson
C. Mervin Palmer I46.
Clarence M. Tarzwell 1*7.
Curtis L. Newcombe I48 •
William J. Tucker l49.
John L. Mohr 50.
William M. Ingram
Peter Doudoroff
William G. Spence
George Paine
Gustave Prevost
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USE AND VALUES OF BIOASSAYS
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DERIVATION OF THE THRESHOLD VALUE FOR TOXICITY AND
THE EQUATION OF THE CURVE BY A GRAPHICAL METHOD
G. E. Burdick
Senior Aquatic Biologist
New York State Conservation Department
Albany, N. Y.
The curvature found in graphs of toxicity when time is plotted against
concentration on logarithmic paper is introduced by the failure of the effec-
tive range to conform to the axes of the paper.
For many years workers in the field of fish toxicology have been
plagued by the curvilinear nature of the graphs of data that cover a wide
range of concentrations. The equation must then be formulated by means of
multiple degree equations, or, alternatively, the data must be transformed
so it may be fitted somewhat approximately by a straight line.
Many methods have been suggested to avoid the necessity of consider-
ing the data as curves, such as the use of the reciprocal of time (Powers,
1917, and a number of English authors); the use of only that part of the data
conforming most closely to a straight line (Herbert and Merkens 1952, and
others) and the conventional plotting as powers, roots, or natural logarithms.
In their TLm method Hart, Doudoroff and Breenbank (1945) avoid the
production of a curve by the use of single values for specified times. How-
ever, in their estimation of biologically safe concentrations, which involves
the relationship between the 24 and 48-hour TLm an approximate straight
line relationship appears to be assumed.
Shepard (1955) straightened his data on the tolerance of brook trout to
low oxygen by applying as a corrective the time for death at zero oxygen.
In most cases concentrations have been plotted as logarithms and con-
sideration of the spread of the data when times for death of individual fish
are noted, also indicates time to be logarithmically distributed. When so
plotted a curve results which usually can be fitted only approximately by mul-
tiple degree equations.
This observation led to an analysis of various methods of plotting and
the procedures commonly used to straighten curvature. As a result of this
study the hypothesis which was stated at the start of this paper was formu-
lated.
Time would not permit the complete discussion of the reasoning and
calculations which led to this supposition and the suggested method for graph-
ical translation. This has previously been prepared and submitted to the
N. Y. Fish and Game Journal for publication.
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Briefly, concentration can affect time only in that range which lies
between the threshold value of concentration and a minimum time for death
which is independent of actual concentration. Only if both of these ap-
proached zero would the data conform to the axes of the paper. If the values
were known, translation of the data could readily be accomplished by sub-
traction. This is not generally feasible.
Usually the minimum time for death at any concentration is small and
if there is sufficient spread, this correction becomes negligible at low con-
centration. Progressive subtraction of different assumed values from the
plotted curve in this range will produce a straight line, which is extended to
cover the entire spread of the data. Only one such line can be produced on
logarithmic paper and the concentration subtracted when it is formed repre-
sents the threshold value. Undercorrection produces a curve in the same
direction as the original, overcorrection a curve in the reverse direction. If
the subtraction is then continued over the remainder of the curve, the points
gradually deviate from the straight line and approach the original curve. The
space between the straight line and this produced curve represents the minimum
time for death. The straight line represents a translation of the data to the axes of
the paper. In order to use this method the data must include the part of the curve
having maximum inflection.
The procedure also provides a method of deriving an equation representing
accurately the original plotted curve. The fit on those curves tested appears
better than that given by the use of multiple degree equations, while the labor
is obviously much less. The equation of the straight line is first found by any
of the usual methods , In this equation, the log (x - the threshold value) is
substituted for log X and the log (y - the minimum time) is substituted for log Y.
This then represents the equation of the original data and closeness of fit can be
determined by substituting selected values of x and solving for y.
While the data to which it has been applied appear to confirm the hypothesis,
it cannot yet be considered completely proved, since it should be applied to many
toxicity curves . The indications are that it may prove a useful tool for the determii
tion of threshold concentrations, since these are derived not as extrapolated values ,
but as a function of the plotted curve. Use of the straightline relationship and
replotting or re-use of the corrected data would appear also to open the way for
the use of a number of statistical analyses which could not be applied to curvilinear
data.
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Literature Cited
Hart, W. B. » Peter Doudoroff and John Greenbank (1945)
The evaluation of the toxicity of industrial wastes, chemicals
and other substances to fresh-water fishes. The Atlantic Re-
fining Co. , Philadelphia, Pa. 317 pp.
Herberts D. W. H. and J. C. Merkens (1952)
The toxicity of KCN to rainbow trout. Jour. Exp. Biol., 29:632 -649.
Powers, Edwin B. (1917)
The goldfish as a test animal in the study of toxicity. 111. Biol.
Monographs, 4:No 2.U. of 111. , Urbana, 111. 73.
Shepard, M. F, (1955)
Resistance and tolerance of young speckled trout (Salvelinus
fontinalis) to oxygen lack, with special reference to low oxygen
acclimatization. J. Fish. Res. Bd. of Canada, 12 (3): pp. 387-446.
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INTERIM PLAN* FOR STANDARDIZING THE
BIOASSAY OF PARALYTIC SHELLFISH POISON BY
USE OF A REFERENCE STANDARD
DEPARTMENT OF HEALTH, EDUCATION AND WELFARE
Public Health Service
Washington 25, D. C.
INTRODUCTION
Prevention of poisoning due to eating toxic shellfish has been
a problem of concern to health and fishery authorities for many years.
On May 26, 1955, the Public Health Service sponsored a confer-
ence to discuss recent developments in the assay for shellfish toxi-
city. The principal objective of the conference was establishment of
a uniform procedure for bioassay of shellfish poison.
State and Federal agencies having representatives at the Confer-
ence included: Department of the Army, Fish and Wildlife Service,
Food and Drug Administration, Massachusetts Department of Public
Health, Canadian Department of National Health and Welfare, State
of New York Conservation Department, and U. S. Public Health Ser-
vice.
The most significant development in shellfish poison assay was
reported by Dr. E. J. Schantz, Chief, Chemistry Branch, Army
Chemical Corps biological research laboratory. Dr. Schantz and
his associates were engaged in this field of research for several years
and succeeded in isolating the poison in pure form. In working with
the purified poison they also found certain color reactions, such as
the Jaffe and Benedict-Bahre tests, which could serve as the basis of
chemical assay for the poison. The conferees agreed that: (1) puri-
fied poison should be used as a tentative reference standard, and (2)
results of future bioassays should be reported in terms of weight of
poison.
The Army Chemical Corps has provided the Public Health Service
with a limited quantity of purified shellfish poison for initiating standard-
ization of the assay procedure. This plan outlines the manner in which
the Public Health Service will distribute the reference standard to lab-
oratories interested in standardizing their bioassay procedures.
* This plan has been developed jointly by representatives of the Public
Health Service and the Chemical Corps. Among those primarily
responsible for preparing the technical details are Dr. E. J. Schantz,
Chief, Chemistry Branch, Fort Detrick, Md.; Mr. E. T. Jensen,
Acting Chief, Shellfish Sanitation Section, Milk and Food Program,
Washington, D. C., Dr. K. H. Lewis, Chief, and Mr. E. F.
McFarren, Chepaist, Milk and Food Research, Robert A. Taft Sani-
tary Engineering Center, Cincinnati, Ohio. The background data
discussed by Mr. McFarren at the seminar will be presented in a
series of papers to be published elsewhere.
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SECTION A
OBJECTIVES
Primary objectives as stated at the 1955 Conference on Shellfish
Toxicology are: (1) National and international standardization of
laboratory techniques used in assay of paralytic shellfish poison, and
(2) determination of the weight of purified shellfish poison equivalent
to one mouse-unit. Schantz and associates have found that approxi-
mately 0. 2 of purified poison is equivalent to one mouse-unit. This
relationship needs to be established in each laboratorv that employs
the reference standard.
Secondary objectives are: (1) Accumulation of data which might
be used to evaluate the statistical reliability of the bioassay, and
(2) development of a chemical test to supplement or replace the bio-
assay.
SECTION B
DISTRIBUTION OF REFERENCE STANDARD
Laboratories interested in utilizing the purified shellfish poison
for standardizing the bioassay procedure should direct their requests
to Public Health Service, Washington 25, D. C. , Attention: Shell-
fish Sanitation Section. Requests from Canadian laboratories can be coi
sidered only when endorsed by the Food and Drug Directorate, Depart-
ment of National Health and Welfare, Ottawa.
In making application for the reference standard the laboratory
director agrees:
(1) To use this material only for standardization of assay
procedures used in connection with control or research
activities on paralytic shellfish poison.
(2) To make no secondary distribution of the reference stanr
dard to other laboratories except those which are under his
administrative control.
(3) To include in the standardization of the assay procedures
the methods recommended herein.
(4) To furnish the Public Health Service with the accumulated
standardization data and typical results of assays employ-
ing the standardized procedures.
The Public Health Service will send participating laboratories
5 to 10 ml of a reference standard containing 100 v^g of poison per ml
in an acidified aqueous solution. Each lot of solution will be assayed
prior to mailing. Additional supplies of the standard are contingent
upon receipt of data noted in item (4) above. Copies of a standardized
report form (See page 30)are available from the Public Health Service.
Investigations of paralytic shellfish poison will be continued
by the Public Health Service. Emphasis will be given to improvement
of chemical assay methods which may supplement or replace the
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presently used bioassay techniques.
Requests for information or consultation on technical problems
related to utilization of the reference standard or development of
assay procedures should be directed to the Robert A. Taft Sanitary
Engineering Center, Cincinnati 26, Ohio, Attention: Chief, Milk
and Food.
SECTION C
OBLIGATIONS OF PARTICIPATING LABORATORIES
Participating laboratories have three obligations under this
program:
(1) Determination of the weight of poison per mouse unit
for their own laboratory's conditions. When this relation has
been established, assay results can be reported in terms of micro-
grams of poison per 100 grams of shellfish meats.
(2) Use the purified poison as a periodic check on their
operating procedures. (Experience to date indicates that CF valued 1
should not deviate from the mean by more than +_ 20%. A recheck
is recommended if this value is consistently exceeded).
(3) Fulfill the agreement noted in Section B.
SECTION D
PROCEDURE FOR BIOASSAY OF REFERENCE STANDARD
SOLUTION
(1) Select healthy mice weighing 19 to 21 grams from the
stock to be used for routine assays. If mice weighing less than
19 grams or more than 21 grams are used a correction factor
must be applied to obtain the true death-time. 2/
(2) Place one ml of the Standard Reference Solution in a
100 ml volumetric flask or graduated cylinder and dilute to 100 ml
with water. If kept at 3 to 4° C this solution is good for several
weeks.
(3) Dilute aliquots of the above solution with distilled water
until intraperitoneal injection of 1 ml doses into a few test mice
causes the median death time to fall between 5 and 7 minutes.
The following dilutions are suggested as a guide:
Parts of poison solution Parts of water ng poison per ml
10 10 0.500
10 15 0.400
10 20 0.330
10 25 0.286
10 30 0.250
1 / CF = *8? poison per ml
mouse unit poison per ml
2J See page 29 for weight correction factors.
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Preparation of smaller increments of dilution, as indicated by
these preliminary tests, will be necessary to obtain satisfactory
results, For example, when 10 parts of poison solution in 25 parts
of water kill the initial test mice in 5 to 7 minutes additional dilu-
tions should be tested which contain 10 parts of poison solutiqn in
24 parts of water and 10 parts of poison solution in 26 parts of
water. The pH of the dilutions should be between 2 and 4 for assay
and must not be higher than 4. 5.
(4) Use 10 mice on each of the 2 dilutions --preferably 3
dilutions-- that fall within the 5 to 7 minute median death period.
Give a 1 ml dose to each mouse by intraperitoneal injection and
determine the death time as the time elapsed from completion
of the injection until the last gasping breath of the mouse.
(5) Repeat the assay one or two days later on the dilutions
as prepared under (3) above.
(6) Repeat the entire procedure starting with step' 1.
(7) Take the median death time of the ten mice for each
dilution within each of the 4 trials and determine the number of
mouse units contained in one ml of each dilution from the Sommer
Tables (page 11). Divide the micrograms of poison in one ml by
the mouse units in one ml. The result is a conversion factor
(hereafter termed the CF value) expressing the micrograms of
poison equivalent to one mouse unit. Compute the average of the
individual CF values and use this average value as a reference
point to check routine assays. This CF value will vary from one
laboratory to another depending on differences in animals and
techniques. The individual CF values may be expected to vary
significantly within a laboratory if techniques and mice cannot be
rigidly controlled. The latter situation would require the con-
tinued use of the reference standard or a secondary standard
depending upon the volume of assay work undertaken by the laboraT
tory.
SE C T IO N E
PROCEDURE FOR USING THE REFERENCE STANDARD
WITH ROUTINE ASSAYS OF SHELLFISH PRODUCTS
The conversion factor (micrograms of poison per mouse
unit), determined as indicated in Section D-7 and termed the CF
value, is used to calculate the micrograms of poison in a sample
of shellfish by multiplying the number of mouse units found in
100 grams of sample by the CF value.
A periodic check on the CF value should be made as follows:
If shellfish products are assayed less than once each week, a check
run should be made each day assays are performed by injecting
5 mice with the reference standard. If assays are made on several
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days during a week only one check need be made each week. The
check run should be carried out on a dilution of poison such that
the median death time falls between 5 and 7 minutes. The _£DF value
thus determined should check the average CF value within - 20%.
If it does not check within this range complete a group of ten mice
by adding 5 more mice to the 5 mice you have already injected, and
inject a second group of 10 mice with the same dilution of poison.
The CF value determined for the second group should be averaged
with that of the first group and the resulting value taken as the new
CF value for the conversion of mouse units to micrograms of poison.
A variation of more than 20% represents a significant change in the
response of the mice to the poison, or in technique of assay. Any
changes of this type should be compensated by a change in the CF
value.
Repeated checks of the CF value ordinarily produce con-
sistent results within the limits prescribed above; if wider variations
are encountered frequently, the possibility of uncontrolled or unrecog-
nised variables in the test procedure should be investigated before
proceeding with routine assays,
SECTION F
PROCEDURE FOR BIOASSAY OF CLAMS OR MUSSELS FOR
SHELLFISH POISON 3/
(1) Preparation of Sample
a. Shucking
Thoroughly clean the outside of the shellfish
with fresh water. Open by cutting the adductor muscles.
Rinse the inside with fresh water to remove sand or
other foreign material. Remove the meat from the
shell by separating the adductor muscles and the tissue
connecting at the hinge. Do not use heat or anaesthetics
preparatory to opening the shell, and do not cut or
damage the body of the mollusk at this state of the pro-
cedure. Collect the meats in a glazed dish until about
100-150 grams of material are obtained.
b. Draining
As soon as possible after shucking, transfer the
shellfish meats to a ten-mesh sieve without layering
and allow to drain for 5 minutes. Discard the drainings.
c. Grinding
Grind the meats in a meat grinder of the house-
hold type which has 1/8 to 1/4 inch holes in the grinding
face, or macerate in a blender until a homogeneous
mixture is obtained.
3/Modilication of a procedure adopted November 19» 1943, by the
U.S. Public Health Service, U.S. Fish and Wildlife Service, and
the Federal Food and Drug Administration, with excerpts from
a procedure used by the Laboratory of Hygiene, Department of
National Health and Welfare, Canada.
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Extraction
a. Weighing
Weigh out 100 grams of the well mixed material
into a tared beaker.
b. Acidification
For all species of clams add 100 ml of 0. 1 N
hydrochloric acid, for sea mussels add 80 ml of
0.1 N hydrochloric acid and 20 ml of distilled water.
Stir thoroughly.
c. Digestion
Heat the mixture and boil gently for 5 minutes,
remove from the heat and allow to cool to room
temperature.
d„ pH adjustment
Adjust the cooled mixture to pH 4. 0 to 4. 5 as
determined by B. D. H. Universal Indicator, brom
phenol blue, congo red paper or a pH meter. To lower
the pH add 5 N hydrochloric acid drop by drop with
stirring. To raise the pH add 0. 1 N sodium hydroxide
dropwise with constant stirring to prevent local alka-
linization and consequent destruction of the poison.
After adjustment of pH make the volume up to 200 ml.
e. Clarification
Stir the mixture to homogenity and allow to
settle until a portion of the supernatant liquid is trans-
lucent and can be decanted off free of solid particles
large enough to block a 26 gauge hypodermic needle.
If necessary it may be centrifuged (5 minutes at 3000
r.p. m„ ) or filtered through filter paper. It is neces-
sary to obtain only enough liquid to carry out the bio-
assay.
Mouse Test
a. Inoculation
Select mice weighing between 18 and 22 grams,
if possible, and never over 25 grams. The mice
should be of the same strain as used in the standardi-
zation procedure (Sections D and E). Inoculate each
test mouse intraperitoneally with 1 cc of the acid ex-
tract obtained in F-2 above. Note time of inoculation
and. observe mice carefully for time of death as indi-
cated by the last gasping breath. Record time by
means of a stop watch or clock with a sweep second
hand. One mouse may be used for the initial deter-
mination but 2 or 3 are preferred. If the death time
or median death time of several mice is less than
5:00 minutes, make a dilution so as to obtain death
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times between 5 and 7 minutes. If the death time of
one or two mice injected with an undiluted sample
is greater than 7 minutes but one or more mice do
die, a total of at least three mice need to be inocu-
lated in order to determine the toxicity of the sample
with confidence. If large dilutions are necessary, the
pH of the dilution should be adjusted by addition of
dilute hydrochloric acid (0, 1 or 0.01 N) dropwise so
that the pH is between 3, 5 and 4. 5 (and never higher
than 4. 5). Inoculate three mice with the dilution that
gives death times between 5 and 7 minutes {death
determined as the time of the last gasp).
b. Calculation of Toxicity
Determine the median death times4/ of the mice,
and from Sommer's table determine the "number of
mouse units corresponding to the median death time.
If test animals weigh less than 18 grams or more than
22 grams, a weight correction must be made for each
mouse by determining the mouse units corresponding
to the death time for that mouse from Sommer's
table, multiplying this value by the correction factor
for that mouse from Sommer's table, and then deter-
mining the median mouse unit for the mice. To deter-
mine the amount of poison per 100 grams of meat mul-
tiply the median mouse-unit by the dilution factor used
in obtaining this unit and then by 200, since the origi-
nal clam extract was made up to 200 ml. Convert the
mouse units to micrograms of poison per 100 grams
of meat by multiplying by the CF value.
4/ Include survivors in determination of median death time.
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TIME - DOSAGE RELATIONS J)'UK FAKAL x no xoru
POISON (ACID) 5/
TIME
DOSE
TIME
DOSE
WEIGHT OF MICE
DOSE
5:00
1. 92
10 gm.
0. 50
05
1. 89
10-1/2
. 53
1:08
100
10
1. 86
11
. 56
10
66. 2
15
1. 83
11-1/2
, 59
15
38. 3
20
1. 80
12
. 62
20
26. 4
30
1. 74
12-1/2
. 65
25
20. 7
40
1. 69
13
. 675
30
16. 5
45
1. 67
13-1/2
. 70
35
13. 9
50
1. 64
14
. 73
40
11. 9
6:00
1. 60
14-1/2
. 76
45
10, 4
15
1. 54
15
. 785
50
9. 33
30
1. 48
15-1/2
. 81
55
8. 42
45
1. 43
16
. 84
2:00
7. 67
7:00
1. 39
16-1/2
. 86
05
7. 04
15
1. 35
17
. 88
10
6. 52
30
1.31
17-1/2
. 905
15
6. 06
45
1. 28
18
.93
20
5. 66
8:00
1. 25
18-1/2
. 95
25
5. 32
15
1. 22
19
. 97
30
5. 00
30
1. 20
19-1/2
. 985
35
4. 73
45
1. 18
40
4. 48
9:00
1. 16
20
o
o
o
i-H
45
4. 26
30
1. 13
50
4„ 06
10:00
1.11
20-1/2
1.015
55
3. 88
30
1. 09
21
1. 03
3:00
3. 70
11:00
1. 075
21-1/2
1. 04
05
3. 57
30
1. 06
22
1. 05
10
3. 43
12:00
1. 05
22-1/2
1. 06
15
3. 31
13
1. 03
23
1. 07
20
3. 19
14
1. 015
23-1/2
1. 075
25
3. 08
24
1. 08
30
2o 98
15
o
o
o
i-H
24-1/2
1.085
35
2. 88
25
1. 09
40
2. 79
16
0.99
26/27
1. 10
45
2. 71
17
0. 98
28/29
1. 11
50
2. 63
18
0. 972
30
1. 12
55
2. 56
19
0. 965
4:00
2. 50
20
0. 96
05
2. 44
21
0. 954
10
2. 38
22
0. 948
15
2. 32
23
0. 942
20
2. 26
24
0. 937
25
2. 21
25
0. 934
30
2. 16
30
0. 917
35
2. 12
40
0. 898
40
2. 08
60
0. 875
45
2. 04
50
2.00
55
1. 96
5/ These data are for IP injections as determined by Dr. Sommer,
and are based on the data furnished by him to workers in the
field.
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FORM FOR REPORTING ASSAY DATA ON REFERENCE STANDARD
SHELLFISH POISON SOLUTION
Log Number Reference Standard or RSS*
Laborato ry
Date of Assay
Date Reference Standard Solution was prepared or received
Pate pf preparation of 1 to 100 dilution of RSS*
Name of Assayer
Strain of mice
Data on individual mice
Dilution
1
2
3
g ppison per ml
pH of dilution
Mouse No,
Mouse
Weight (^m)
death time
(seconds)**
death time
(seconds)**
death time
(seconds) **
1
•
%
. 3
4 .
5
...v vt ...... .i. i
6
7
8
9
19
•' " n J 1 ¦ I1 '•»' ' "" ' ' " '
Median death time***
Mouse units per ml
Send 3 copies of data sheets to: Sanitary ]
tngineering Center,
Cincinnati 26, Ohio Attn; Chief. Milk 5md Food.
*RSS" Reference Standard Solution.
** J?eath time from completion of injection to last gasp.
***In elude survivors in determination of median death time.
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APPLICATION FACTORS TO BE APPLIED TO BIOASSAYS
FOR THE SAFE DISPOSAL OF TOXIC WASTES
Croswell Henderson
ROBERT A. TAFT SANITARY ENGINEERING CENTER
U. S. Public Health Service
Cincinnati, Ohio
The tremendous expansion of the chemical industry, especially
that dealing with petrochemicals, the newer metals, synthetic fibers,
insecticides, and detergents, presents new and difficult problems in
the protection of aquatic life from effects of industrial pollution.
Necessary chemical methods for the detection and measurement of
many substances in these complex wastes have not been developed.
Even when chemical methods are available, toxicity information gen-
erally is not, nor can toxicity always be attributed to one or more sim-
ple materials. In complex wastes there may be a number of different
toxicants or there may be synergy or antagonism between substances
so that the toxicity of final effluents may be entirely different from that
of its components.
By using bioassays correctly, the effect on aquatic life of complex
industrial wastes may be determined directly. Relatively simple bio-
assay methods have been devised and are being currently used. The
procedures recommended by the subcommittee on toxicity of the Sew-
age and Industrial Wastes Federation (1) which are based in part on
research previously reported by Hart, Doudoroff, and Greenbank
(2) are quite satisfactory.
The basic bioassay procedure consists essentially of preparing
different concentrations of an effluent or other test material with a
selected dilution water, adding the test fish and observing their reac-
tions over a definite time period. A logarithmic series of concentra-
tions is generally most convenient.
For effluents of unknown toxicity, it is desirable to make explora-
tory or small scale tests to determine the approximate toxic range.
Two fish are added to 2 liters of each test solution over a wide range
of concentrations; e. g. , 100, 10, 1, and 0. 1 percent effluent. Obser-
vations over a short time period will indicate the necessary concentra-
tions for the full scale experiments.
For the full scale tests, it is desirable to use a minimum of ten
fish for each test concentration. This may be conveniently done in
five gallon wide mouth glass bottles, using five fish in ten liters with
duplicate samples. A series of intermediate concentrations are set
up in the range indicated by the exploratory tests. For example, if
the exploratory tests indicate an effect on fish between 10 and 1 per-
cent, concentrations of 10, 5. 6, 3. 2, 1. 8, and 1. 0 percent, or in
some cases intermediates between these are set up and fish added to
each.
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The dilution water used should be water from the receiving stream
above the effluent outlet, if suitable for fish, or water of similar
characteristics, particularly with respect to pH, alkalinity, and hard-
ness.
The test fish should be a species adaptable to laboratory condi-
tions such as temperature, feeding and handling, should be of relatively
small size and readily available. Fathead minnows (Pimephales
promelas) and bluegill sunfish (Lepomis macrochirus) from 1-1/2 to
2-1/2 inches long and weighing from 1 to 2-1/2 grams have been satis-
factory for work in several laboratories. Many other species may be
suitable. In many cases it is preferable to use a species native to
the receiving water or at least one directly comparable.
The above tests are designed so that no oxygenation or aeration
is generally necessary. Atmospheric absorption of oxygen by the
exposed water surface adequately takes care of the requirements of
the fish during the test period. However, if strong concentrations of
high oxygen demand effluents are being tested, oxygen or air may be
needed. Such methods as solution renewal at definite time intervals,
introducing air or oxygen at a specific rate and maintaining an oxygen
interface over the solution in the test jar, have been used successfully.
Physical and chemical determinations such as temperature, dissolved
oxygen, pH, alkalinity, acidity, hardness, etc. , are made periodi-
cally during the course of the bioassays.
Observations as to the fish reactions are generally made for a
96 hour period, twenty-four, 48, and 96 hour TLm (median tolerance
limit-concentration which causes 50% mortality) vaines are estimated
through straight line graphical interpolation.
Example - 5. 6% - All fish dead in 48 hrs.
3. 2% - 3 of 10 fish survive 48 hrs.
1. 8% - 7 of 10 fish survive 48 hrs.
1. 0% - All fish survive 48 hrs.
Controls - All fish survive 48 hrs.
10
7—
3 —
8
% Survival
1
10 20 30 40 50 60 70 80 90 100
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Once the TL value has been obtained, the next step is to
determine how tc?^"use it. Obviously a concentration that will kill
50% of a test species is not safe for aquatic life. Liberal applica-
tion factors (sometimes erroneously called safety factors) must be
applied. Methods of applying laboratory bioassays have received
very little attention. Industry and others are now using bioassays
to some extent and more are ready to use them if provided a rela-
tively simple bioassay procedure and reasonable methods of apply-
ing the results.
Stepwise, a proposed method of applying bioassays to meet
specific industrial waste problems is as follows:
(1) Perform laboratory bioassays to obtain TLm values.
(2) Calculate the dilution water required.
Example: 48 hour TL -2. 4%
m
Effluent flow - 10 cfs
Dilution Ratio - 2.4:97.6 or 1:40
10 cfs effluent would require 400 cfs dilution water from
the receiving stream to produce a condition which would kill half
of the test fish in 48 hours.
(3) Develop and apply a numerical factor that will provide
safety to aquatic life in the receiving water. This can be in the
form of a whole number when applied to dilution water needed or a
fractional value of the TLm when applied to the effluent.
There are many individual factors to be considered in develop-
ing a single application factor for specific wastes. The major ones
may be grouped as follows:
A. Relating laboratory bioassays to actual conditions.
(1) Conversion f rom 50% to 100% survival.
(2) Conversion of acute or short term toxicity to possi-
ble long term or chronic effects of a toxicant.
(3) Conversion of toxicity in non-renewed or static
test solutions to continuous flow conditions.
B. Relating test fish to other aquatic life.
(1) Effect on test fish versus other more sensitive fi^h
species that may be present in the receiving stream,
(2) Other stages in the life cycle such as fry or eggs
may be more sensitive.
(3) Some of the major fish food organisms may be more
sensitive.
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C. Variability in the toxicity of effluents which may fluctuate
considerably over a period of time. Bioassays with 24
hour composite samples of effluents from various indus-
tries over a three month period, have produced the follow-
ing results:
48 hr. TL % Concentration
m
Type of No . of
Industry Samples
Petrochemical
Oil Refinery
By products
coke
Sewage plant
(containing indus-
trial wastes)
Chemical & dye 1
'* 2
3
2
4
6
6
Maximum
Toxicity
2.6
37
3.7
3.3
4.4
13.5
Minimum
Toxicity
22
40
13.5
22
16
28
Average
Toxicity
9.6
38.5
9.0
10.2
11.9
20.0
3 7
4.2
24
8.5
It is evident that any one sample taken for bioassay at a
particular time may give erroneous results. Either a large
number of samples would have to be tested or some factor
developed to provide for conditions of maximum toxicity. The
above factors are generally of concern in all cases and are
not provided for in the experimental procedures .
D. Other individual factors may have considerable effect on
toxicity but are generally provided for in the tests. Any
major differences between test and actual conditions in
the' following characteristics must be considered.
(1) Change in water quality characteristics
Temperature and Dissolved oxygen - Toxicity
is generally'greater at higher temperatures and
lower dissolved oxygen.
pH, Alkalinity, Hardness - These characteris-
tics are somewhat interdependent. The buffering
capacity would determine the pH which may have an
effect on the toxicity.
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Example - The toxicity of metals, cyanides, ammonia,
and many other toxicants is greatly influenced by pH
and other characteristics.
Metals; Cyanides
^m
% Cone.
pH - Alk - Hardness
(2) Synergy and antagonism - A combination of materials
may produce a more toxic or less toxic effect. It has
been reported that copper and zinc in the same solution
will produce an effluent eight times more toxic than
copper alone. Likewise a material such as calcium may
have an antagonistic effect (3).
The addition of several different effluents to a receiving water
may give an entirely different toxicity picture from that of the separ-
ate effluents. For example, one industry may release an effluent
containing metals into a hard water stream. The metals may pre-
cipitate or complex into insoluble compounds with materials in the
stream waters and become relatively non toxic to aquatic life. Fur-
ther downstream, another industry may release an acid effluent,
lowering the pH and causing the metal to go into solution and become
highly toxic. Bioassays with receiving waters would indicate the
toxicity but the toxic materials may be attributed to the wrong indus-
try. For this and othgr reasons, a knowledge of the chemical com-
position of the effluents and what reactions may be expected are of
extreme importance.
If the above factors are carefully examined and related to estab-
lished experimental evidence, sizeable application factors are appar-
ent. An application factor of 10 times the 48 hour TL__ has been
tentatively suggested. Circumstances in a particular study may raise
or lower this value. Some individual factors, as already mentioned,
may add to the above figure. Others which may detract some from
the magnitude of the figure and must be considered are natural puri-
fication, loss of volatiles, oxidation or hydrolysis to non toxic pro-
ducts, precipitation, complexing, antagonism, etc.
The following example may illustrate a means of arriving at an
application factor, providing the necessary information is available
or can be obtained:
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Factor
How Derived
Numerical Value
A. Test vs Actual conditions
B. Test fish vs other aqua-
tic life
C. Variability of effluents
D. Other variables
(see page 34)
Experiments 2
Research
Experiments 3
Research
Experiments 2
Research
Provided for in tests
Application Factor = 2x3x2 = 12
Using the previous example of dilution water required for 10
cfs of an effluent having a 48 hr. TLm value of 2. 4%.
400 x 12 = 4800 cfs dilution water required for safety to aquatic
life.
It is evident that considerable research is necessary before
definite factors can be assigned which will be accepted. Good experi-
mental evidence and a definite basis for the derivation of these fac-
tors will have to be available. Factors should be suggested which
will adequately do the job and yet not cause undue expense fox treat-
ment.
Several methods of approach to this problem are suggested.
The laboratory fundamental research program may give some defi-
nite answers or at least methods of mathematical expression. These
could consist of long time continuous flow experiments with different
wastes or waste components using various species of fish, different
life history stages, and other aquatic organisms. The information
could be mathematically related to our short term static bioassay.
Other fundamental knowledge needed which could be obtained is the
effect on toxicity of changes in water quality characteristics such as
turbidity, temperature, dissolved oxygen, pH, alkalinity, hardness,
etc. Programs to obtain this information could be and to some ex-
tent are being carried out here at the Sanitary Engineering Center,
as well as in states, universities, industrial laboratories, and other
research institutions.
Another approach which may give partial but more immediate
answers is the field approach of working with actual effluents under
field conditions. Bioassays could be made of the effluents from a
particular industry or group of industries. Operating conditions,
effluent flows, stream flows, and other conditions could be obtained.
A study of the biota of the receiving waters could detect what is
happening under present operating conditions. Study of several
plants connected with a particular type of industry may give evidence
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as to the magnitude of the application factor needed. The infor-
mation may also be valid for related industries with similar types
of effluents.
An ideal situation for a study of this type would be one con-
tinuously operating plant on a stream which may lagoon or regulate
waste flows. The effect on the biota may be determined at different
flow rates and the application factor estimated which affords pro-
tection to aquatic life.
These problems can be worked on for an extended period and
still no mathematically precise answer obtained as with certain
physical and chemical phenomena. However, a reasonably accu-
rate estimate may be obtained. With proper application factors
conditions may be estimated which are reasonably safe for aquatic
life. How satisfactory they are can only be determined by studying
the stream biota after a period of operation when they are in effect.
There are many ways in which industry can use bioassays.
Toxicity of final effluents and the probable effects on receiving waters
can be determined. Toxic components can be traced in process
effluents, which may permit the treatment of much smaller quanti-
ties of waste. The effectiveness of treatment processes may be
establiahed. In the location of new plants, the amount and quantity
of dilution water necessary or the degree of treatment of wastes
may be ascertained in advance of construction.
It is believed that the greatest use will be made of bioassays if
the methods are relatively simple and easy to use. Reliable and
suitable application factors should be developed.
References
1. Doudoroff, P., et al. , "Bio-Assay Methods for the Evaluation
of Acute Toxicity of Industrial Wastes to Fish. " Sew. and Ind.
Wastes, 23, 11, 1380-1397, (1951).
2. Hart, W. B. , P. Doudoroff, and J. Greenbank, "The Evaluation
of Industrial Wastes, Chemicals, and Other Substances to Fresh-
Water Fishes. " Waste Control Laboratory of the Atlantic Re-
fining Company, (1945).
3. Doudoroff, P. , and M. Katz, "Critical Review of Literature on
the Toxicity of Industrial Wastes and Their Components to Fish.
II. The Metals, as Salts. " Sew. and Ind. Wastes, 25, 7, 802-
839, (1953). —
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USE AND VALUE OF BIOLOGICAL- INDICATORS OF POLLUTIpN
-38-
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SOME PROBLEMS IN THE IDENTIFICATION OF MICROORGAMISMS
Wm. Bridge Cooke
ROBERT A. TAFT SANITARY ENGINEERING CENTER
U. S. Public Health Service
Cincinnati, Ohio
In studying the populations of algae and fungi on high-rate and stand-
ard-rate trickling filters recently, it was found that within groups of well
known organisms, such as algae, growing in this habitat there were pro-
blems of identification which were difficult if not impossible to solve
using preserved materials. For instance, in the case of a coccoid green
alga, would you feel confident in determining from a pickled sample whethe:
the cells belonged to Chlamydomonas, Chlorococcum, Protococcus,
Palmellococcus, some other coccoid genus, or were only early palmelloid
stage cells of Stigeoclonium? Under certain conditions of observation
after preservation of a sample in the field any one or several of these
names could be applied to a collection. In the case of blue green filamen-
tous algae, could you tell from a preserved specimen which had been
broken up in a Waring Blendor so that filament length or branching were
not factors in the collection, whether you had Oscillatoria, Phormidium,
Lyngbya, or some other form? Yet, we had such problems to answer.
Several specialists in the field of green algae were consulted on the iden-r
tification of the coccoid organisms, and only after prolonged study and
as a result of familiarity with the groups did G. W. Prescott come up
with an answer which admittedly was only tentative. Dr. Drouet was sent
material of the filamentous blue-green, which turned out to be an Amphi r
thrix. ———
This experience led me to the idea that it might be well to develop
a discussion of the problem of the identification of organisms that it is
h,oped may be used as indicative of pollution by their presence or absence
in a stream. Some of the pitfalls in the identification of such organisms
may be mentioned.
In the taxonomy and systematics of any group of organisms, plant or
animal, there are usually two schools of thought. In the colloquial lan-
guage of the trade of taxonomy these are referred to as the splitters
and the lumpers. Modern taxonoxny has shown that there is room for
^oth, but in the past the matter has had the finality of either/or. On the
basis of minute variations, spme species of organisms have been split
into many species, while without visible differences but with microscopic
variation not visible to a casual observer, species have been lumped to-
gether.
In the early years of systematic biology the cataloging of the animals
and vegetation pf the earth's surface was of prime importance. As time
went on, it became important to integrate the studies so begun, and for
various regions floras and faunas were produced; lists, annotated or not,
of various organisms were developed; and monographic treatments of
genera, families, orders, and even classes were attempted. As mountaina
of materials were built1 up'in the herbaria, museums, and culture collectipn
-39-
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of the world, more and more such synthesizing of our knowledge couid
be endeavored. Much of this work was done with the assumption,
stated or implied, that certain barriers existed between species, barriers
which prevented exchange of genetic materials between populations. When
such barriers were based on political lines, as in the older works and
the earlier catalogues, they were unnatural and the assumption of separ-
ate populations invalid within certain groups. Depending on the members
of the populations, many geographic barriers may be considered valid,
others not.
Where specimens were not available to a student of a group, where
reliance had to be placed on records regardless of reliability, the devel-
opment of generic monographs without regard to unnatural barriers
resulted in burdensome lists of species in excess of numbers which could
be supposed with little effort to by synonymous. Where specimens were
available and critical interpretation of parallel geographic species was
carried out, the lists became shorter. Where undue emphasis was
placed on minute detail as valid bases for species separation, lists of
species became quite lengthy. Where minute detail was considered a
jtHtrsequifcur for species criteria but the result of individual variation,
species lists shrank. Based on specimens of leaves from one tree, a
species has been based on the shade leaves of the common beech, another
on the sun leaves.
In the genetics laboratory, an elementary problem is occassionally
presented in which the common fruit-fly, Drosophila melanogaster, is
used. A group of females of one type is mated with a group of males of
another. The types are chosen with the idea of developing the concepts
involved in the statistical study of Mendelian ratios. The progeny of
such crosses are obviously different from or similar to their parents.
Yet if a taxonomist of the pre-Mendelian school were to observe these
he could easily find the bases for several species of Drosophila, if not
for new genera.
These introductory remarks are used to indicate that as our know-
ledge of life and our ability to classify its various aspects progress with
the increasing knowledge from various fields such as morphology, cytol-
ogy, anatomy, physiology, genetics, etc. , the concepts of species and
other categories change. As we find more and more specimens of or-
ganisms, some of which appear to vary markedly from known species,
our concepts of various groups change. Various myths of taxonomy
become exposed as additional materials are obtained. For instance, Dr.
W. M. Ingram mentioned recently that many species of Hawaiian snails
were shown to have been invalidly proposed when it was discovered that
the basic character, that of color of shell, was found to have been al-
tered for gain by those paid to collect additional color variants.
The use of dried specimens of plants, skins of animals, prepared
mounts of micro-fauna, dried and otherwise prepared materials of algae,
fungi and bacteria, has its place in modern taxonomy. At one time these
were the only type of material on which concepts of species were based.
More and more, modern taxonomy is using such materials as "type" ma-
terials or as records of past accomplishment and historical fact. The
type specimen is the basis on which the species is founded, but the living
specimen is the basis on which species in many groups of organisms are
-40-
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best known and studied today. The geneticist needs living material for
various types of genetic manipulation, as do the physiologist, the experi-
mental morphologist, and the other scientists who base their work on
the types of activity used by the first three mentioned. The systematic
botanist today more and more uses genetic techniques for proving or
denying the existance of one species or another in developing generic
monographs. Experimental taxonomy is being actively used in many
fields of botanical and zoological classification as well as in bacteriologi-
cal and mycological systematics. In many cases, not only taxonomy
and genetics are being learned in this way, but, at the same time and
with the same organisms, many bits of information useful in "pure" and
"applied" science are being discovered.
The present state of systematic identification of micro- and macro-
organisms in the average field laboratory is based largely on well estab-
lished texts in which a limited number of organisms are described and
illustrated. Such texts may be considered comparable to a mushroom
handbook in which only the most obvious form is illustrated with a photo
taken more or less recently or a painting or a pencil sketch made more
or less accurately within the last half century. Such description and
nomenclature as may be used do not take into consideration the wide vari-
ation possible within the species, nor do they consider the fact that two
species of mushroom obviously identical to the naked eye may be quite
different both in respect to edibility and to micromorphology. The pro-
cess of revising a genus of bacteria, fungi, algae, protozoans, mollusks,
diatoms, or any other group, is long and laborious. Unless it is a genus
which includes species of great ecdnomic importance such as Penicillium
or Aspergillus , the work must be carried out in the "ivory tower" of '
the University or the Academy, otherwise it will interfere with or con-
flict with the interests of the employer whose business is the practical
application of one process or another, of one species or another, or the
development of one product or another.
In the development of a taxonomy which is adequate to the needs of
the student of the biological indicators of pollution, not only at the level
of specialization represented by the members of this panel but also at
the level of usability of the man in the field, many factors will have to be
tfiken into consideration, Not the least of these is the development of
monographic studies at the genus or family level, in which by use of the
techniques of experimental taxonomy a species may be given a more
qound definition comprehensible to the man in the field whether he be ;
biologist, engineer, or chemist, for it is rare that all three professions
)vill be represented adequately on any field staff. Such dynamic rather
than static taxonomic studies should be carried out in all fields in which
organisms occur that may be found under varying conditions and types
of pollution. The studies should overlap into other fields, since organ-
isms occurring in or prevented from occurring in polluted waters may
also be of importance in agriculture, forestry, food processing, indus-
trial production, and other fields. Information from the laboratory
should be available concerning the pollution tolerances of such species.
Following the production of the monographic studies, the information
contained therein of interest to the pollution biologist should be gathered
together and made available to him in language understandable by a
relatively unspecialized college graduate.
-41-
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best known and studied today. The geneticist needs living material for
various types of genetic manipulation, as do the physiologist, the experi-i
mental morphologist, and the other scientists who base their work on
the types of activity used by the first three mentioned. The systematic
botanist today more and more uses genetic techniques for proving or
denying the existance of one species or another in developing generic
monographs. Experimental taxonomy is being actively used in many
fields of botanical and zoological classification as well as in bacteriologi-
cal and mycological systematics. In many cases, not only taxonomy
and genetics are being learned in this way, but, at the same time and
with the same organisms, many bits of information useful in "pure" and
"applied" science are being discovered.
The present state of systematic identification of micro- and macro-
organisms in the average field laboratory is based largely on well estab-
lished texts in which a limited number of organisms are described and
illustrated. Such texts may be considered comparable to a mushroom
handbook in which only the most obvious form is illustrated with a photo
taken more or less recently or a painting or a pencil sketch made more
or less accurately within the last half century. Such description and
nomenclature as may be used do not take into consideration the wide vari-
ation possible within the species, nor do they consider the fact that two
species of mushroom obviously identical to the naked eye may be quite
different both in respect to edibility and to micromorphology. The pro-
cess of revising a genus of bacteria, fungi, algae, protozoans, mollusks,
diatoms, or any other group, is long and laborious. Unless it is a genus
which includes species of great ecdnomic importance such as Penicillium
or Aspergillus , the work must be carried out in the "ivory tower" oi T
the University or the Academy, otherwise it will interfere with or con-
flict with the interests of the employer whose business is the practical
application of one process or another, of one species or another, or the
development of one product or another.
In the development of a taxonomy which is adequate to the needs of
the student of the biological indicators of pollution, not only at the level
of specialization represented by the members of this panel but also at
the level of usability of the man in the field, many factors will have to be
taken into consideration, Not the least of these is the development of
monographic studies at the genus or family level, in which by use of the
techniques of experimental taxonomy a species may be given a more
§ound definition comprehensible to the man in the field whether he be .
biologist, engineer, or chemist, for it is rare that all three professions
yrill be represented adequately on any field staff. Such dynamic rather
than static taxonomic studies should be carried out in all fields in which
organisms occur that may be found under varying conditions and types
of pollution. The studies should overlap into other fields, since organ-
isms occurring in or prevented from occurring in polluted waters may
also be of importance in agriculture, forestry, food processing, indus-
trial production, and other fields. Information from the laboratory
should be available concerning the pollution tolerances of such species.
Following the production of the monographic studies, the information
contained therein of interest to the pollution biologist should be gathered
together and made available to him in language understandable by a
relatively unspecialized college graduate.
-41-
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Such studies, properly presented, should enable a pollution biologist
to interpret readily the role of the more important organisms in the
stream, sewage treatment plant, or possibly even water treatment plant
under his jurisdiction, in terms of work load or indicator value of which
those organisms are capable, much as the forester can determine the
possibilities of his forest in terms of board feet of lumber available and
potential crop replacement.
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USE AND VALUE OF BACTERIAL INDICATORS OF POLLUTION
Paul Kabler
ROBERT A. TAFT SANITARY ENGINEERING CENTER
U. S. Public Health Service
Cincinnati, Ohio
Normal Intestinal Bacteria
Following Pasteur's epic work and Kock's discovery of solid media,
rapid progress was made in the isolation and description of the bacter-
ial species commonly found in the intestinal tract.
S. typhosa (B. typhosus) described by Eberth in 1880.
Vibrio cholera described by Kock in 1884.
E. coli (Bacillus coli) described by Escherich in 1886.
It was clearly shown that E. typhosa, V. cholera, and the Salmonellae
were associated with enteric disorder s~~and it was hoped that suitable
procedures for the identification of these and other etiological agents
pf human disease might be developed for use as indices of drinking water
quality. Several ingenious techniques were developed and remain with
certain modifications as valuable research tools. There are many instan-
ces on record in which specific pathogenic bacteria have been isolated
from suspected water sources, however, such attempts almost always
ended in failure when used as a routine test for water quality. It is, there
fore, apparent that attempts to isolate pathogenic microorganisms from
a water supply are not recommended in the routine examination as an
index of pollution for the following reasons:
1. Present available methods are tedious, laborious, and require
high technical skill.
2. By the time a pathogen is recovered from a water the harm has
long been done.
Subsequent to the demonstration by Escherich that E. coli was a
normal intestinal organism, numerous investigators showed that E. coli
and related organisms were consistently present in sewage, polluted
streams, lakes and wells and in contaminated soil. These data showed
that:
1. Coliform organisms are normally present in the feces of all
warm blooded animals.
2. Some coliforms are to be found on plants and grains.
3. Feces contain from 5 to 500 million coliforms per gram, about
60 to 95% of which are E. coli.
4. A. aerogenes usually numbers 10, 000 to 500, 000 per gram of
feces.
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5. Coliform organisms are rarely found in virgin soil, but may be
numerous in cultivated soils. From 65 to 80% may be A. aero-
genes.
6. Coliforms may survive weeks or months in fresh water, the pre-
dominating type varying with environmental conditions.
7. Coliforms may live in dry soil 45 to 100 days, but in moist soil
may survive a year or more.
Because the coliform group is constantly present in alimentary dis-
charges, and because of the comparative ease of enumerating them, the
coliform organisms have become the accepted indicator of fecal pollution
Also because of the fact that water containing fecal pollution may contain
intestinal pathogens, the "coliform test" has assumed importance as a
criterion in judging the sanitary quality of water.
The Coliform Group
"The coliform group shall include all of the aerobic and facultative
anaerobic Gram-negative nonspore-forming bacilli which ferment lac-
tose with gas formation in 48 hrs. at 35° C. " (Standard Methods for the
Examination of Water and Sewage, 10th edition, p. 375).
The first coliform test was devised at the New York State Depart-
ment of Health Laboratories by Theobald Smith in 1893 using dextrose
broth and first edition of "Standard Methods" was published in 1905.
The current edition (10th) of Standard Methods provides for no differ-
entiation of "fecal" and "nonfecal" types. It is stated, "Such differenti-
ation is of little moment in determining the suitability of water for humai
consumption, as contamination with either type of waste renders the
water potentially unsatisfactory and of unsafe sanitary quality". Current
methods have certain recognized imperfections as to time required,
specificity and reproducibility. Continuous efforts have been directed
toward development of better methodology. Some improvements have
been:
1. Substitution of lactose for glucose in the presumptive medium,
which materially reduced the number of presumptive false posi-
tive tests.
2. Churchman's observation that some dyes will selectively inhibit
certain species or group of Gram-positive and of spore-forming
organisms.
3. Modifications of incubation times, amount of gas, media ingredi-
ents and incubation temperatures.
a. Bile salts by MacConkey (1908).
b. Perry and Hajna modified Eijkmann test (1933).
c. McCrady, brilliant green bile (1937).
d. Mallmann, lauryl sulphate tryptose broth (1941).
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The adoption of brilliant green lactose bile broth as a confirmatory
medium is perhaps the most important advance in methods in the last
fifty years. The use of lauryl sulfate tryptose broth in the presumptive
test is also an important improvement.
Recently suggested (tentative method only) membrane filter tech-
niques may prove to have far reaching applications.
Other Organisms as Pollution Indicators
Streptococci
A leading "pretender" as an indicator of pollution is the Strepto-
coccus (Enterococcus).
The Streptococci as a group have wide distribution. Many of them
are pathogenic for man and animals. Some strains are strict parasites
while others are saprophytic in existence. Many are fastidious in their
growth requirements while others will grow with ease.
The sewage Streptococcus or Enterococcus have been proposed as
an indicator organism on the basis that:
1. They are present in feces and sewage and are found in known
polluted waters.
2. They are not found in pure waters, virgin soil and sites out of
contact with human and animal life.
3. They do not multiply outside the animal body (except in such
media as milk). (Suckling, Exam. Waters & Water Supplies
[1943] , Blakiston & Co. )
There is incomplete agreement on these three points. Evidence indi-
cates the Enterococci do not multiply in water. Whether they persist
longer in water or disappear at about the same rate as coliforms is a
disputed point.
The sewage Streptococci are never present in as large numbers as
coliforms. The Streptococcus coliform ratio variesfrom 1/100 down to
1/10. As the Streptococcus detection methods are apparently no more
sensitive than coliform methods, and as the pollution density is less,
this would have the practical effect of reducing the sensitivity of the
method.
Streptococcus classification methods have attempted to distinguish
between human and animal strains. Mannitol-positive strains "do ap-
pear more common in human feces and raffinose-positive strains more
common in bovine feces, " but separation on this basis has no place
in evaluating the sanitary quality of a water supply.
Streptococcus methods offer a promising line of investigation. At
this time the Streptococcus index in not a satisfactory substitute for
the coliform group. Its greatest value is as a corroborative test where
the coliform datum is suspected.
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The Anaerobes
Spore-forming, lactose-fermenting anaerobes have been used
as indicator organisms in water. The anaerobic organism is usually
referred to as Clostridium welchii in England and Clostridium per-
fringens in America.
The spore-forming Clostridium is unsuitable as an indicator or-
ganism for the following reasons:
1. They are extremely resistant to destruction and survive for
long periods.
2. They are abundant in animal manure, cultivated soil, and de-
composing organic material.
3. They fail to correlate with results of sanitary surveys.
4. There are small differences in their numbers in heavily pol-
luted and pure waters and they give no evidence of the degree
of pollution.
The major interest in anaerobes at present centers about the
occasional false positive presumptive test and its carry-over to bril-
liant green bile broth tube with gas and decolorization. The anaerobe
can be eliminated by its failure to grow in formate ricinoleate broth.
The problem of finding a better indicator organism is one of para
mount interest. Some show promise, but none at present compete
with the fifty odd years of experience with the coliform estimation
and its correlated data.
Escherichia coli
Originally Bacterium coli was considered to be a single species.
As new methods and tests were devised, this so-called single species
was divided into more and more species. Now this group of organ-
isms is considered under the collective term of "coliform organisms"
Hundreds of research workers have added their contributions to
the studies to determine the characteristics of the bacteria included
in the coliform group, so that the ideal bacterial criterion of pollu-
tion might be defined. From this mass of data has evolved a classi-
fication of the coliforms into the coli group, the intermediate group,
and the Aerobacter group. These differences are not sharp, well-
defined HisHncHoni, but are rather differences that shade from one
variety to another on the basis of the interpretation of results from
multiple testing procedures. It is further complicated by the question
of what intermediate varieties to include with the E. coli as pollution
indicator organisms. — ~
Lewis and Pittman found no significant difference in the ratio of
E. coli to coliforms in polluted water and in water of high sanitary
quality. Ruchhoft, et al. , found the ratio of E. coli to coliforms in
a sewage effluent and at the pumping station fo be about the same
-46-
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although the coliform M. P. N. counts were 13, 815 and 0. 219 per milli-
liter, respectively. Taylor found the E. coli ratio of no help in esti-
mating the sanitary quality of water in "English lakes.
A summation of the evidence of many workers warrants the follow-
ing conclusions.
1. The ratio of E. coli to that of other coliforms is not an index
of water quality.
2. Presence of E. coli usually is no more significant than other
members of tEe coliform group.
3. The additional labor involved in the routine identification of
E. coli in the presence of other coliforms is not justified on
the basis of the information attained.
4. The sanitary quality of water is dependent on the total number
of coliform organisms present. Density is an indication of
pollution.
Summary
From a bacteriological point of view the coliform group is currently
the best available indicator for use in the estimation of pollution of
waters and in the sanitary evaluation of pure waters of potable quality.
The coliform test is a quantitative test. Significance of its interpreta-
tion is dependent on determining the density (or most probable number).
Only when the count exceeds a normally expected minimum number of
coliform organisms is pollution indicated. Few samples of water are
completely free from coliforms when a sufficiently large sample is
examined. The established standard of the Public Health Service
for drinking water recognizes the possible occurrence of an occasional
coliform.
In the foregoing discussion it has been evident that pathogenic organ-
isms cannot be used as indicator organisms. Their unsuitability is
based on infection of the public before their discovery, their abnormal
occurrence in the feces of a small percent of any group, and their small
number in comparison to other intestinal type organisms.
It is freely admitted that there are normally found in the intestinal
tract of man many types of organisms not belonging to the "coliform
group". Some of these organisms at various times have been considered
as "indicator organisms". The available data eliminated each for one
or more reasons.
The substitution of E. coli for the present coliform group is unsuit-
able "because it gives no additional information for the increased labor
and because there are not sufficient epidemic studies to interpret the
E. coli index.
Butterfield stated that . while it is desirable to seek for better
bacterial criteria of pollution in water, and these researches should be
continued, the results of such studies to date have not produced a criter-
ion which may be considered to have satisfactorily replaced the coliform
group. " This summation is equally true today.
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1,
2.
3.
4,
5.
6.
7.
8.
9
10
11
12
13
14
Bibliography
Smith, T. A New Method for Determining Quantitatively the Pollu-
tion of Water by Faecal Bacteria. Thirteenth Annual
Report of the State Board of Health of New York for
1892, 712 (1893).
Standard Method of Water Analysis. First edition. Amer. Pub.
Health Assn, (1905)
McCrady, M. H. A Practical Study of Procedures for the Detec-
tion of the Presence of Coliform Organisms in Water.
Amer. J. Pub. Health, _27:1243 (1937).
Mallmann, W. L. and Darby, C. W. Uses of Lauryl Sulphate
Tryptose Broth for the Detection of Coliform Organisms.
Amer. J. Pub. Health, 31:127 (1941).
Perry, C. A. and Hajna, A. A. A Modified Eijkmann Medium.
J. Bact. _26:419 (1933).
MacConkey, A. T. Bile Salt Media and Their Advantages. J.
Hyg. 8:822 (1908).
Ruchhoft, C. C., Kallas, J. G. , Chinn, B. and Coulter, E. W.
Coli-aerogenes Differentiation in Water Analysis. J.
Bact. 21:407 (1930), 22:125 (1931).
Ruchhoft, C. C. Studies on the Longevity of Bacillus typhosus
in Sewage Sludge. Sew. Wks. J. 61 1054 (1934).
Parr, L. W. Viability of Coli-aerogenes Organisms in Culture
and in Various Environments. J. Infectious Dis. 60:
291 (1937).
Parr, L. W. The Occurrence and Succession of Coliforn Organ-
isms in Human Feces. Am. J. Hyg. 27:67 (1938).
Taylor, C. B. The Ecology and Significance of the Different Types
of Coliform Bacteria Found in Water. J. Hyg. 42:23
(1942). ~
Clark, H. F. , Geldreich, E. E. , Jeter, H. L. and Kabler, P. W.
The Membrane Filter in Sanitary Bacteriology, Pub.
Health Rpts. 66^:951 (1951).
Geldreich, E. E. , Kabler, P. W. , Jeter, H. L. , and Clark,
H. F. A Delayed Incubation Membrane Filter Test for
Coliform Bacteria in Water. Amer. J. Pub. Health
45:1462 (1955).
Stokes, E. J. , Jones, E. E. and Miles, A. A. Effect of Drying
and Digestion of Sewage Sludge on Certain Pathogenic
Organisms. Abs. Sew. Wks. J. 17: No. 6, 1302
(1945).
-48-
-------
15, Mom, C. P. and Schaeffer, C. O. Typhoid Bacteria in Sewage
and Sludges. Sew. Wks. J. 12:715 (1940).
16, Winter, C. E. and Sandholzer, L. A. Recommended Procedure
for Detecting the Presence of Enterococci. Commercial
Fisheries Bull. T. L. 2. U.S. Fish and Wildlife Ser-
vice (Nov. 1946).
17, Mallmann, W. L. , Seligmann, E. B. A Comparative Method for
the Detection of Streptococci in Water and Sewage.
Amer. J. Pub. Health, 40:286 (1950).
18, Litsky, W. , Mallmann, W. L. and Fifield, C. W. A New Method
for theDetection of Enterococci in Water. Am. J. of
Public Health, 43:873 (1953).
19, Litsky, W. , Mallmann, W. L. and Fifield, C. W. Comparison
of the Most Probable Numbers of E. coli and Entero-
cocci in River Waters. Am. J. I^ub. Health 45:1049
(1955). —
?0. Slanetz, L. W. , Bent, D. F. and Bartley, C. H. Use of the Mem-
brane Filter Technique to Enumerate Enterococci in
Water. Public Health Repts. 70:67 (1955).
21. Gilcreas, F. W. and Kelly, S, M. Relation of Coliform Organ-
ism Test to Enteric Virus Pollution. J. A- W. W. A.
45:683 (1955).
2?, Butterfield, C. T. Determining the Bacterial Quality of Water.
Reimpreso del Organo Ofici^l de la Asociacioh
Interamericana de Ingenieria Sanitaria. Ano 2,
July, 1948.
no.
1
t9-
-------
PROTOZOA AS INDICATORS OF THE
ECOLOGICAL CONDITION OF A BODY OF WATER
James B. Lackey
University of Florida
Gainesville, Florida
Introduction
The topic as assigned by the program chairman was limited to
protozoa. "Protista" would be a better term for several reasons. One
is that much of this work has been concerned with various species of
colored flagellates, whereas only a few colorless flagellates today are
assigned to the protozoa and even these are regarded as colorless algae
by some workers. Another is that very few ciliates are regarded as
indicator species - some ciliates seem to prefer anaerobic habitats
and others aerobic, but there is little to indicate a marked preference
for pollution by any of them. The third reason is that the Sarcodina
have been far too little investigated in respect to pollution. There is
some indication that a small group of the minute amoebas are charac-
teristic of certain stages of domestic sewage treatment but the larger
naked amoebas are rarely abundant anywhere. Decloitre (1) is cur-
rently tending to show that the thecate amoebas are cosmopolitan, where-
as there has been a tendency to regard most of them as sphagnum bog
types. Foraminifera and Radiolaria we know are marine - the former
in the ooze, the latter pelagic. For these reasons, any discussion
should be more widespread than the colorless flagellates, the ciliates
and the amoebas.
The Varying Nature of Environments
An organism generally occupies a given niche permanently because
that niche offers the most favorable environment for it. Such occupa-
tion implies choice on the part of the organism. It does not take into
account crowding or other competition, changing the environment as
by shutting off light, being forcibly removed or retained as by a current,
etc. In other words, we too frequently assume an environment to be
relatively constant, whereas it is anything but constant and the organisms
in it either must be adaptable to change, or migrate, or die. New kinds,
suited to the new environment, may replace them and our concept must
be flexible to take care of such changes. There was a lake in Florida
which was often referred to as "a good bass lake. " Within one grow-
ing season, it became covered with water hyacinths and the bass disap-
peared, migrating downstream. This did not alter the concept implied
by the phrase "good bass lake, " but it certainly showed that such con-
cepts are applicable to transient situations.
Actually, we talk as if we are stating a constant. Over a broad
environmental range, such relative constants actually exist. Thus
-50-
-------
a coal mine stream, among other things may constantly contain enough
sulfuric acid to be highly acid, a great part of the time. At such times,
it is favorable to a small group of organisms, the most striking being
Euglena mutabilis. This Euglena has been noted as abundant also in
iron seeps where ferric hydroxide was concentrated and is occasionally
found in a variety of other situations, some of them alkaline. What
favors Euglena mutabilis in the acid coal mine stream? Does it have
a greater physiological tolerance to sulfuric acid than does, say
Euglena polymorpha? Or is there less competition for food and sun-
light? Or is there a particular food substance in such a stream?
Until we have answers to some such questions as these, we are on
dangerous ground in talking of indicator organisms. Euglena poly -
morpha is one of several species of this genus which increase markedly
in streams receiving effluents from sewage treatment plants, It has
been referred to as an indicator species of recent fecal pollution.
But it also occurs as dense blooms in certain types of swamp waters,
such as the cedar swamp at Woods Hole, Massachusetts. These two
situations must have in common some other factor than fecal pollution.
Ubiquitous Organisms
Some organisms tolerate such a wide range of environmental
variation as to almost defy identification of limiting factors. Man is
the most conspicuous of these. In our case, we can adapt to extremes
of heat, cold, moisture, dryness, pollution of the atmosphere, and
so on through a wide range. The blue crab, Callinectes sapidus,
occurs along the Atlantic coast from Main to the Florida Keys, as well
as the Gulf Coast - a geographic range within which there is a wide
fluctuation of temperature, food, enemies, salinities and other factors.
In Florida it migrates up the rivers and into some of the fresh springs,
or those almost fresh. But what of the small ciliate, Cyclidium
glaucoma, which apparently is truly cosmopolitan and which Finley
has shown (2) can transfer from fresh to sea water or vice versa?
It rarely attains huge numbers anywhere but in a hay infusion it may.
The only indicator value it has in such a situation is to indicate that
the bacteria on which it feeds are very abundant and that competition
for this bacterial food is at a low ebb. This is easily amenable to
experimental proof - one has only to grow Cyclidium with and without
competing predatory ciliates. ~
The number of «uch ubiquitous species may be large. Undoubtedly,
we often fail to recognize them. For example, Didinium nasutum,
rarely common anywhere, but used by Beers (3) as a fresh water '
experimental ciliate, is frequent in samples from the high seas. Some
Vorticella species belong in this category also and apparently
Chlorella among the green algae. Peranema trichophorum and Anison-
ema ovale of the colorless Euglenophyceae' are others.
Organisms of Somewhat Limited Distribution
It seems an easy matter to make a long list of organisms which
occur in either salt or fresh water but not both. Or, for marine
-51-
-------
TABLE I
Probable
Oxygen
Relationship
Of Species
Rhizopoda
Nuclearia delicatula
Dimastigamoeba gruberi
Arpioeba vespertilio
" radiosa
Pelomyxa palustris
Vahlkampfia limax
" albida
" guttula
" fragilis
Hartmanella hyalina
Chlamydophrys stercorea
Microgrdmia. sp.
Arcella vulgaris
Cochliopodium bilimbosum
Euglypha a,lveolata
Diplophrys archeri
Mastigophora
Anthophysa vegetans
Bodo caudatus
11 several other species
Bicoeca sp.
Cercobodo spp.
Clautriavia parva
Hexamitus inflatus
" crassus
0)
ao
nt
*
v
CO
IX
X
X
X
X
X
X
X
-52
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Species
Of Frequent Occurrence or May
Occur in Large Numbers in
s»
A
U
V *>
A 5
0 ?
•H 4)
s *
4) TJ
4)
CD 4_>
)h nt
0) 0)
tj U
CO <*¦>
£ £
1 *
o u
«°
2 Tt
5
nt -•->
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
a
(U
o
•H
u
C
<0
s
r c
> n)
o
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
a
S.S
t *
2
a
o
0«
T3
4)
N
U C
o
31
"1
41 h
JS &
•H
i-H
* CO
a
x
X
X
nt
*
•X3
V
O
a,
a
&
X
X
X
X
X
X
X
X
-------
Mastigamoeba spp.
X
X
X
Mastigella spp.
X
X
X
Helkesimastix faecicola
X
X
X
Monas spp.
X
X
X
X
X
X
Oicomonas spp.
X
X
X
X
X
X
Pleuromonas jaculans
X
X
X
X
X
X
Tetramitus spp,
X
X
X
X
Spiromonas angusta
X
X
X
X
Desmarella moniliformis
X
X
X
X
Cladospongia elegans
?
X
Gyromonas ambulans
X
X
X
Cercomastix parva
X
X
X
Poteriodendron petiolatum
X
X
X
X
X
Sterromonas formicina
X
X
X
Phanerobia pelophila
X
X
i
X
Trigonomonas compressa
X
x
X
X
Trepomonas spp.
X
X
X
X
Urophagus rostratus
X
X
X
X
Volvocales
Carteria spp.
X
1 X
X
X
X
X
Chlamydomonas spp.
X
; x
X
X
X
X
Chlorogonium spp.
X
: x
X
X
X
X
Dunaliella salina
X
X
Eudorina elegans
X
X
X
X
X
Gonium pectorale
X
X
X
X
X
11 sociale
X
X
X
X
X
Pandorina morum
X
.
X
X
X
Polytoma uvella
X
X
X
X
Polytomella citri
X
X
X
X
Platymonas spp,
X
X
X
X
Chlorobrachis
X
X
X
(Pyrobotrys gracilis)
Spondylomorum quarter-
X
X
X
X
X
X
X
X
X
X
X
X
narium
Astasia spp.
X
X
X
X
Entosiphon sulcatum
X
X
X
X
Notosolenus spp.
X
X
X
X
X
Peranema trichophorum
X
X
X
X
Euglenophyceae - green
Cryptoglena pigra
X
X
X
X
Euglena acus
X
X
X
X
11 agilis
X
X
X
X
" deses
X
X
X
X
" gracilis
X
X
X
X
X
" mutabilis
X
" pisciformis
X
X
X
X
X
" polymorpha
X
X
X
X
X
" quartana
X
X
X
X
" sanguinea
X
X
X
" tripteris
X
X
X
X
" viridis
X
•
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
-53-
-------
Eutreptia viridis
JLepocinclis butschlii
" ovum
" texta
Phacus parvulus
" triqueter
" pyrum
Trachelomonas urceolata
11 volvocina
Chrysophyceae
Botryococcus braunii
Chromulina ovalis
" globosa
Mallomonas spp.
Coccolithophora
Pontosphaera sp.
Dinoflagellata
Oxyrrhis marina
Gymnodinium splendens
Prorocentrum triangu-
latum
Cryptophyceae
Chilomonas paramecium
Cryptomonas spp.
Rhodomonas spp.
Ciliata
Carchesium sp.
Chilodonella spp.
Cinetochilum margari-
taceum
Colpoda spp.
Colpidium sp.
Glaucoma scintillans
Halteria grandinella
Lionotus faeciola
Metopus sigmoides
Opercularia spp.
Oxytricha spp.
Paramecium spp.
Saprodinium putrinum
Stylonichia spp.
Trimyema compressa
Urocentrum turbo
Vorticella spp.
Hastatella radians
Enchelyomorpha vermi-
cular is
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X '
X
X
X
X
X
X
X
X
X
X
X
1
X
X
X
X
X
f
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
54-
-------
species, pelagic versus neritic species, as ivauiuiaiiu r. j.
certain dinoflagellates. Even boreal, termperate and tropic species
may occur: Chrysococcus cingulum is common at Woods Hole and
the Chesapeake but has not been found in three years' examination
of Florida Gulf Coast samples. Certain species of ciliates, such as
Irimyema compressa and Enchelyomorpha vermicularis seem
restricted to water - or mud - devoid of oxygen, but are widespread
if enough such situations are examined.
In this second category, it is always difficult to determine the
limiting conditions. Metopus (possibly more than one species) is
often found along with the two ciliates above but will exist also in the
presence of some oxygen. In these instances, is it a lack of oxygen
or the presence of H^S which is effective? In the Radiolariais it
the ions of full strength sea water or merely physical osmotic pheno-
mena? The chloromonad Gonyostomum semen is common to the
brown waters of cedar and cypress swamps but is almost unknown
elsewhere. There are many instances of limited distribution but
knowledge of the limitations is virtually lacking.
Rare Organisms
Many species are named in the literature which the average
observer has never seen. This may mean poor observation, not
having sampled the habitat type for the organism, or it may really
be rare. The diatom Attheya zachariasi has frustules of such
transparency that they are seen only with considerable difficulty and
its chloroplasts and cytoplasm are usually grouped in a small flat-
tened disc. It is seldom reported in the United States, yet seems
common. Another is Cephalomonas granulosa, one of the green
flagellates. This distinctive organism occurs sparingly in Ohio
Valley waters but on one occasion, a slight bloom of them was re-
corded in L.ytle Creek (4); the species was originally described from
Maryland (5). Trachelomonas reticulata is colorless and Klebs (6)
described it from "faulenden" cultures, so it should occur in polysa-
probic situations. There are no known records of the species in the
United States, except a very dense bloom found by the writer in an
Alabama tree hole. A fourth rare organism is the ciliate, Hastatell<
radians. It is large (100 |jl) and unmistakable but has been seldom
encountered. On April 8, 1956, it was the dominant ciliate, there
being 80 per ml, in a deep sink hole in Orange Lake, Florida. The
sink hole acted as a concentrating pit for the slowly drying up lake,
there being a slow but steady outflow through the 71 foot deep bottom
Undoubtedly then, many organisms which we consider rare, wi]
attain high numbers if the environment is favorable. The question
generally develops into attempts to recognize the optimal factors.
Discussion
Before stating a relationship between organism species or
organism numbers and pollution, we must first define the pollutant.
There are many kinds of pollution, varying from an excess of organ
-55-
-------
or putrescible matter to heavy concentrations of inorganic salts.
Poisons, even physical conditions, enter the picture. It is hardly
worthwhile to list types of pollution here but there are chemical and
physical tests which are revealing even when a condition is not
apparent to the eye. Conditions having been determined, we ascertair
the numbers and kinds of organisms present and the permanence of
both the physico-chemical condition and the population.
The mere occurrence of an organism in a sewage treatment
plant does not mean it is indicative of pollution - Oxytricha may oc-
cur either in a sewage treatment plant or in a hay infusion simply
because it finds in each place the bacteria it uses as food, as well as
ample oxygen. A long list of organisms occurring in sewage treat-
ment plants could be made and the same organisms would be found
in other situations in many cases. Table I is such a list in part. It
does not cover the green nonflagellated algae and the diatoms. It
is based partly on personal experience and partly on the known litera-
ture. No references are given, for there would be too many and in
addition, the list does not cover all the references which could be
made. It is weakest in regard to the occurrence of ciliates.
This table is intended to show that there are few organisms which
are common to only one type of habitat. It does not include those
which are common to or which sometimes occur in huge numbers in
waters not known to contain substances we would regard as pollutants
or fertilizers. Nor does it include laboratory cultures. Undoubtedly,
there are enriching substances in the gravel pits around Chillicothe,
Ohio, which annually bloom with Uroglena, or the Cedar Swamp at
Woods Hole which annually blooms with Gonyostomum. But at present
we either do not know what they are, or we do not regard them as
pollutants. The same is true of the Gulf of Mexico water in which
Gymnodinium brevis blooms so heavily, or the once famous Spiro-
stomum pool at Woods Hole. The vast majority of waters will
have many species in common and often one or more of these species
will bloom. But the organisms do not classify such waters; hence,
fall outside the scope of this discussion.
It cannot be too strongly emphasized that a cause and effect
relationship does not necessarily exist simply because of abundance
of an organism and occurrence of a defined pollutant. Clark (7)
has described mechanical trapping of plankton in certain situations
and has also called attention to common orientations and attractions
of organisms to others of their own kind. "Trapped" populations
may remain in a situation, so we cannot be too sure of even this
yardstick. Statements as to the indicator value of an organism in
a given situation absolutely demand critical experimental analysis
and accumulation of much data. Even then the relationship to pollu-
tion may be secondary. The outfall of the Tampa sewage treatment
plant apparently has little effect on chlorophyll bearers in its section
of Tampa Bay but the number of tintinnid ciliates is considerably
higher here than in other sampled sections of the Bay. Counts of the
-56-
-------
bacterial population have not been made but an increased numoer mere
could account for the increased ciliates. The magnitude of the sewage
contribution and the size of the Bay are discouraging to attempts at
collecting precise and continuous data. It would be necessary not
only to secure tidal flow data, but seasonal and vertical biological
data here and elsewhere in the Bay, then add chemical analyses. At
the end of this costly process, it is questionable where relationships
would have been established. In November, there is a bloom of the
diatom, Skeletonema, but it varies in density from year to year; it
also occurs in other portions of the Bay which receive no sewage.
There are repeated dense but local blooms of many dinoflagellates,
such as Gymnodinium splendens, Ceratium furca and others in
various parts of "tampa Bay, but no explanation of why they occur.
Many thousands of dollars have been spent on attempts to determine
the cause of the Red Tide organism, Gymnodinium brevis. So far,
all it indicates is trouble.
Still there are some hopeful signs. Study of the algae or proto-
zoa in a fairly constant environment reveals some which tend to occur
permanently. Yount (8) concluded that no one factor determined the
species density in a habitat. He worked in the Silver Springs
(Florida) boil area or near it, where many factors are uniform the
year round. Even so, light and some other factors fluctuate and
since they worked with diatoms this is important. Such seemingly
small matters as the surface of a slide soon after immersion, as
compared to the same film-covered slide after several days can and
do make surprising differences as to what and how many diatoms
are found on the slide. There has been too little work of this type
but it has been experimental experience that one way to learn what
species of colorless Euglenophyceae mayTbe found in a habitat, is
to suspend slides for several days.
Studying smaller streams indicates that the more abundant is the
oxidizable matter, or the inorganic salts, the more varied and abun-
dant is the suspended population. Lytle Creek (Ohio) studied at
five stations and Cowan Creek, nearby, and studied at one station
(4) over a period of three months in the summer showed 167 and 89
species respectively. Two of the Lytle Creek stations were in a
polluted area, receiving the effluent from a badly overloaded sewage
treatment plant. In this area, certain types of Euglenophyceae
appeared in large numbers. Others died out, especially most specie
of Trachelomonas. Chrysophyceae also failed to pass through this
polluted zone, although ll species were present above. Apparently,
these are responses to domestic sewage pollution.
Unhappily, most of the Chrysophyceae are also lacking in the
small Santa Fe River of Florida. This stream had a large number
(332) of species during the time it was studied and 99 of them were
the same as those in Lytle Creek. The Santa Fe is unpolluted and
receives little drainage from arable land; total numbers of organism
were low. The same story holds for the two forks of the Licking
River of Kentucky - that fork draining the steep, wooded areas such
as Magoffin County in Eastern Kentucky is fairly rich in species
-57-
-------
but low in numbers of organisms. The more westerly fork, draining
the rich blue grass country, is high in both species and numbers.
After many years, the status of indicator species among the
algae and protozoa seems little near clarification. Immediate sewage
pollution of a stream, resulting in oxygen depletion, tends to elimi-
nate all but a few species of anaerobic ciliates and colorless flagell-
ates. Settling the sewage in oxidation ponds results in heavy growths
of Chlorococcales and Volvocales. Allum (9) says that raw sewage
lagoons in South Dakota showed DO values as high as 370% saturation,
attributable to algae. Silva and Papenfuss (10) made a systematic
study of the algae of sewage oxidation ponds in California and in an
extensive comparative review of the literature recorded some 26
genera from at least 10 of their ponds, which list generally agreed
with lists of other investigators. A few other algae were recorded
and a few Euglenophyceae.
The trouble with observations such as these lies neither in their
accuracy nor in the high counts. Rather, it is the tendency we acquire
to regard the organisms found as characteristic of sewage pollution.
Such is not the case. In the Orange Lake pool referred to above,
many of these same species occurred in great numbers and many of
them are common in situations where there is no sewage or sewage
effluent. In fact, commercial fertilizers will produce heavy growths
of some of them.
Partial sewage treatment, resulting in a sharp decrease in bac-
teria and BOD increases the number of ciliates and colorless flagell-
ates, and in the effluent channels, attached blue green and green
algae, the latter usually two genera. Further reduction of BOD
tends to produce some eight or ten species of Euglena and Phacus
and to increase the species of Volvocales and Euglenophyceae. TKere
is little doubt that certain species of algae and protozoa are readily
associated with sewage pollution. But we should distinguish between
"characteristic of" and "indicative of, " the latter being a more
restrictive term.
This last distinction might help make the suggestion of Beck (11)
work. In it, he adds the numbers of species found which are known
to be associated with clean water to the numbers of species found,
which are known to be associated with polluted water. He says it
should apply only to macroscopic invertebrates found in Florida.
However, a detailed analysis of a stream station usually reveals a
large number of species of algae and protozoa. If we can satisfac-
torily allocate them to one or the other of the above classifications,
the result should classify the stream at the station studied. Alloca-
tion is a difficult task and few workers are in agreement for more than
a very few species.
-58-
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References
1. Decloitre, L. 1952. Recherches sur les Rhizopodes TJiecamoebiens
de l' A. O. F. Thesis presented a la Faculte des Sciences de Marseille
Universite D'Aix Marseille.
2. Pinley, H. R. 1930. Toleration of Fresh Water Protozoa to Increase*
Salinity. Ecology 11(2) pp 337-346.
3. Beers, C. Dale. 1947. The Relation of Density of Population to
Encystment in Didinium nasutum. Jour. Elisha Mitchell Scientific
Soc. 63(2) pp 141-154.
4. Lackey, James B. 1956. Stream Enrichment and Microbiota. In
Press. Public Health Reports.
5. Higinbotham, Noe. 1940. Cephalomonas, A New Genus of the Volvo-
cales. Pub. No. 55. Chesapeake Biol. Lab. Solomonas Island, Md.
8 pp.
6. Pascher, A. 1913. Die Susswasserflora Deutschlands, Osterreichs
und der Schweiz. H. 2.
7. Clark, George L. 1954. Elements of Ecology. John Wiley and
Sons, Inc. New York. XIV + 534 pp.
8. Yount, J. L. 1956. Productivity of Florida Springs. Third Annual
Report to Biology Branch, Office of Naval Research.
9. Allum, M. O. 1955. Lagoon Purification Performance in South
Dakota. The American City. 70. pp 128-29.
10. Silva, Paul C. and George Papenfuss. 1953. A Sjystematic Study
of the Algae of Sewage Oxidation Ponds. State Water Pollution Con-
trol Board No. 7. pp 1-35.
11. Beck, William E. , Jr. Suggested Method for Reporting Biotic
Data. 1955. Sewage and Industrial Wastes. 27. pp 1193-97.
-59-
-------
ALGAE AS BIOLOGICAL INDICATORS OF POLLUTION
C. Mervin Palmer
ROBERT A. TAFT SANITARY ENGINEERING CENTER
U. S. Public Health Service
Cincinnati, Ohio
It appears evident to many workers that particular genera or even
species of algae, when considered separately, are not reliable indicators
of the presence or absence of organic wastes in water. However, when a
number of kinds of algae are considered as a community, that group may
be reliable as such an indicator. Such populations may be useful even
in the determination of the degree of purification which has occurred;
but the data available on this is more limited. Therefore, it has seemed
advisable to omit here any attention to intermediate stages in the natural
purification process.
Emphasis is placed in this article on the algae which various workers
have found to be tolerant to relatively undecomposed sewage or closely
related organic wastes. These algae would, in general, be those found to
be present and relatively prominent in the polysaprobic and alphameso-
saprobic zones in a polluted stream. These terms, however, are used
by relatively few workers who have reported sewage tolerant algae. The
writer has interpreted their findings as they apply to sewage pollution.
Due to the large number of references used, an occasional misinterpreta-
tion should not seriously distort the composite record concerning the
algae.
The reports of fifty-six workers have been examined to date. The
genera and species of algae tolerant to sewage or to related conditions
have been recorded, and a total of more than 500 kinds of algae has been
compiled. The maximum number of authors listing any one genus was
34, and, one species, 18.
In order to tabulate the information, the writer has allotted arbitrary
numerical values to each author's record of an alga. A value of two was
given to each alga reported as very highly tolerant and a value of one, to
each alga highly tolerant to sewage. Lightly tolerant and non-tolerant
algae were not recorded in this compilation. The total points from all of
the authors was then determined for each genus and species. The algae
were arranged in the order of decreasing emphasis byjthe authors as a
whole. The fifty genera and the fifty species of sewage tolerant alga« ,t
with the highest total number of points are given in Tables 1 and Z. Plate
I. "Polluted Water Algae" illustrates nineteen of the fifty genera listed
in Table 1.
-60-
-------
Flagellates containing photosynthetic pigments have been included as
algae along with the blue-green algae, green algae, and diatoms. The
record cannot take into account any inaccurate identifications of the algae
which may have been included by each author nor the variations in the
many polluted and stagnant waters to which the original records refer.
The lists of algae in Tables 1 and 2 are meant to be aids for indivi-
duals engaged in stream pollution surveys or related projects. They give
a general consensus of opinion as to the relative significance of the many
sewage tolerant algae which have, so far, been reported. Particular care
can thus be taken in biological surveys to check for the presence of these
genera and species of algae during the microscopic examination of samples.
Samples which are found to contain, in considerable numbers, several
of the algae which are high in the lists may be interpreted as indicating
water that is polluted or stagnant and with high organic content. The list
of species in Table 2 should be more reliable in this respect than the list
of genera in Table 1, since almost every genus referred to included cer-
tain species which are pollution tolerant and others which are not. This
is particularly true with such genera as Navicula, Synedra, and Chlamydo-
monas. In a few instances this same condition may be true also for a
species.. For example, Fjerdingstand (2) claitns that there are two separ-
ate kinds of Ulothrix zonata, the "pollution type" and the "pure water type. "
Additional records by other workers would undoubtedly change the
relative positions of the algae in both the genus and species lists. This
is particularly so for the algae near the ends of the lists where a relatively
few reports are responsible for their present positions.
Three algae are included in Table 2 for which no binomial is assigned.
These are Chlamydomonas sp. , Scenedesmus sp. , and Spirogyra sp. For
none of these genera was there any one species with a total of five or more
points, although the number of points for each genus was high. Relative
positions were assigned to these by arbitrarily dividing by three the figures
for each of the three genera. Many workers have reported these algae
by genus name only and have not referred to the particular species involved.
Examples of species which have been reported as sewage tolerant are
Chlamydomonas reinhardi, Scenedesmus quadricauda, and Spirogyra com-
munis.
No exhaustive list of the clean water algae has as yet been compiled
by the writer, although a smaller representative group of clean water algae
has been selected and illustrated in color (10). The same illustrations
but in black and white, are included here as Plate II. * The clean water
species, listed by genus names only on the plate, are as follows:
Ankistrodesmus falcatus var. acicularis, Aphanothece stagnina, Calothrix
parietiria, Chamaesiphon incrustans, Chromulina rosanoffi, Chrysococcus
rufesceni"| Cladophora glomerata, Cocconeis placentula, Cyclotella bodanTca,
*The original illustrations in color, were painted by Mr. Harold J. Walter
at the Robert A. Taft Sanitary Engineering Center.
61-
-------
POLLUTED WATER ALGAE
PHORMIOIUM
MERISMOPEDIA
LfcPOCINCLlS
CH;..AMV^OBOTRVS
NIT ZS CHI A
ANABAHNA
EUGLENA
TfT RAEORON
CHLOROCOCCUM
SPIROGYRA
OS OIL I.ATORIA
PHACl.
Chlorogonium
CMLORELLA
GOMPHONt-MA
ST1QEOCLOWUM
G I. OF. CCA PSA
UYNG9YA
CHLAMYOOMONAS
-------
CLEAN WATER ALGAE
'j: ado^hora
CMROMULINA
MICHOCOLEUS
COCCONEIS
-------
Hildenbrandia rivularis, Lemanea annulata, Meridion circulare, Merismo-
pedia glauca, Micrasterias truncata, Microcoleus subtorulosus, Navicula
gracilis, Phacotus lenticularis, Pinnularia nobilis, Rhizoclonium hierogTy-
phicum, Rhodomonas lacustris, Staurastrum punctulatum, Surirella
splendida, and Ulothrix aequalis.
The references given below are limited to the articles which should
be particularly useful in work involving the consideration of pollution
tole rant algae.
References
1. Butcher, R. W. 1949.
Pollution and repurification as indicated by the algae. Fourth
International Congress for Microbiology (held) 1947. Report of Pro-
ceedings .
2. Fjerdingstad, E. 1950.
The Microflora of the River M«Slleaa with special reference to
the relation of the benthal algae to pollution. Folia Limnologica
Scandinavica . No . 5 .
3. Forbes , S. A., and R. E. Richardson. 1913
Studies on the biology of the upper Illinois River. Bull. Illinois
State Lab. of Nat. Hist. 9 (Art. 10):481-574.
4. Kolkwitz.R. 1950.
Oekologie der Saprobien. Uber die Beziehungen der Wasserorganismei
zur Umwelt. Schriftenreihe des Vereins fur Wasser-, Boden- und
Lufthygiene Berlin-Dahlem . Piscator-Verlag Stuttgart.
5. Lackey, J. B. 1941.
The significance of plankton in relation to the sanitary condition
of streams. Pages 311-328 in Symposium on Hydrobiology. Univ. of
Wisconsin.
6. Lackey, J. B. 1956.
Stream enrichment and microbiota . Public Health Reports ,
71:708-718.
7. Liebmann, H. 1951.
Handbuch der Frischwasser- und Abwasserbiologie. R. Oldenbourg,
Munchen.
8. McGauhey, P. H., and H. F. Eich. 1922.
A study of the stream pollution problem in the Roanoke, Virginia,
Metropolitan District. Part 3, Third Portion: The plankton of the
waters and muds . Bull. Va. Polytechnic Institute (Eng. Exo. Sta .
Series No. 51) 35:64-88.
-64-
-------
9. Palmer, C.M. 1932.
Plankton algae of White River in Marion County and Morgan
County, Indiana . Butler Univ. Bot. Stud., 2 :125-131. Relation of
algae to sewage pollution was obtained from unpublished records of
the author.
10. Palmer, C. M., and C.M. Tarzwell. 1955.
Algae of importance in water supplies. Public Works Mae.,
86 (No. 6):107-120.
11. Patrick, R., 1948,
Factors effecting the distribution of diatoms. Bot. Rev., 14
(No. 8)-.473-524.
12. Purdy, W. C . 1930.
A study of the pollution and natural purification of the Illinois
River. II. The plankton and related organisms. U.S. Pub. Health
Bull. No. 198:1-212 .
13. Silva, P. C., and G. F. Papenfuss . 1953.
A systematic study of the algae of sewage oxidation ponds.
California State Water Pollution Control Board. Publ. No. 7.
14. Weston, R . S, and C . E. Turner . 1917.
Studies on the digestion of a sewage-filter effluent by a small
and otherwise unpolluted stream. Contrib. from the Sanitary Res.
Lab. and Sewage Exp. Station. Mass. Institute of Tech., Vol. 10.
15. Whipple, G. C., G. M. Fair, and M. C. Whipple. 1948.
The microscopy of drinking water. 4th ed., J. Wiley and Sons, N. Y
16. Wiebe, A. H. 1927.
Biological survey of the upper Mississippi River with special
reference to pollution. Bull. Bur. Fisheries, 43 (Pt. 2):137-l67.
Document No. 1028.
-65-
-------
Table 1
Pollution Tolerant Genera of Algae
List of the Fifty Most Tolerant Genera
In order of decreasing emphasis by 56 authorities
Genera
No.
Authors
Total
Points *
1.
Oscillatoria
34
57
2.
Euglena
31
53
3.
Navicula
19
32
4.
Chlorella
18
30
5.
Chlamydomonas
17
29
6.
Nitzschia
15
25
7.
Stigeoclonium
18
25
8.
Phormidium
15
22
9.
Scenedesmus
12
18
10.
Synedra
14
17
11.
Arthrospira
9
16
12.
Spirogyra
10
15
13.
Phacus
9
14
14.
Gomphonema
9
14
15.
Melosira
9
12
16.
Pandorina
9
12
17.
Ulothrix
10
12
18.
Lepocinclis
7
11
19.
Lyngbya
7
11
20.
Chlamydobotrys
6
10
21.
Chlorogonium
6
10
22.
Tribonema
6
10
23.
Anabaena
8
10
24.
Spondylomorum
8
10
25.
Carteria
5
9
66*
-------
No. Total
Genera Authors Points*
26. Ankistrodesmus 6 9
27. Hantzschia 7 9
28. Pediastrum 7 9
29. Cladophora 8 9
30. Anacystis 5 8
31. Eudorina 6 8
32. Spirulina 6 8
33. Cyclotella 7 8
34. Fragilaria 7 8
35. Cryptomonas 8 8
36. Cymbella 4 7
37* Micr actinium 4 7
38. Closterium 5 7
39. Stauroneis 7 7
40. Chlorococcum 4 6
41. Merismopedia 4 6
42. Stephanodiscus 4 6
43. Cofcconeis 5 6
44. Cosmarium 5 6
45. Cryptoglena 5 6
46. Gonium 5 6
47. Oocystis 5 6
48. Stiphococcus 5 6
49- Surirella 5 6
50. Trfcchelomonas 5 6
~ Tolerance by author "Very High" - 2 points
Tolerance by author "high" - 1 point
-67-
-------
Table 2
Pollution Tolerant Species of Algae
List of the Fifty Most Tolerant Species
In order of decreasing emphasis by 56 authorities
Species
No.
Authors
Total
Points*
1.
Euglena viridis
18
33
2.
Nitzschia palea
14
23
3.
Oscillatoria limosa
10
15
4.
Oscillatoria tenuis
11
15
5.
Arthrospira jenneri
8
14
6.
Stigeoclonium tenue
9
14
7.
Euglena gracilis
7
12
8.
Chlorella vulgaris
8
12
9.
Oscillatoria formosa
8
11
10.
Phacus pyrum
5
10
11.
Chlamydomonas sp.
(6)
(10)
12. ,
Euglena polymorpha
7
10
13.
Oscillatoria chlorina
7
10
14.
Oscillatoria putrida
7
10
15.
Spondylomorum quater-
narium
7
10
16.
Oscillatoria chalybea
8
10
17.
Phormidium uncinatum
8
10
18.
Chlorella pyrenoidosa
5
9
19.
Gomphonema parvulum
5
9
20.
Oscillatoria lauterbornii
5
9
21.
Euglena oxyuris
6
9
22,
JLepocinclis texta
6
9
23.
Hantzschia amphioxys
7
9
24.
Euglena deses
6
8
25.
Oscillatoria princeps
6
8
*68-
-------
No.
Total
Species
Authors
Points*
26. Pandorina morum
6
8
27. Phormidium autumnale
6
8
28. Anabaena constricta
5
7
29. Chlorogonium euchlorum
5
7
30. Melosira varians
5
7
31. Cryptoglena pigra
6
7
32. Chlamydobotrys gracilis
4
6
33. Euglena pisciformis
4
6
34. Lepocinclis ovum
4
6
35. Merismopedia tenuissima
4
6
36. Navicula crytocephala
4
6
37. Nitzschia acicularis
4
6
38. Scenedesmus sp.
(4)
(6)
39. Synedra ulna
4
6
40. Cyclotella meneghiniana
5
6
41. Euglena intermedia
5
6
42. Stichococcus baciliaris
5
6
43. Oscillatoria splendida
6
6
44. Phormidium foveolarum
6
6
45. Pediastrum boryanum
3
5
46. Spirogyra sp.
(3)
(5)
47. Eudorina elegans
4
5
48. Euglena fusca
4
5
49. Surirella ovata
4
5
50. Ulothrix zonata
4
5
~ Tolerance by author "Very High" - 2 points
Tolerance by author "High" - 1 point.
-69-
-------
Dr. Ruth Patrick was unable to attend the seminar.
However, in order to make our coverage of the subject of
indicator organisms more complete she subsequently kindly
consented to prepare a paper for' inclusion in the Transactions .
Her paper on diatoms as indicators of environmental conditions
is herewith presented.
-70-
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DIATOMS AS INDICATORS OF CHANGES IN ENVIRONMENTAL CONDITIONS
By
Ruth Patrick
Curator of Limnology
Academy of Natural Sciences of Philadelphia
Diatoms, which are one-celled algae belonging to the Chrysophyta,
have long been of interest to students of the microscope. The earliest
students of these plants were concerned with the structure of the
siliceous cell walls and the general morphology. Diatoms were often
used as test objects for lenses in order to determine the ability of a
lens to define a given structure. One of the first to become interested
in the geographical distribution of diatoms was Ehrenberg. In 1829
he published a paper on the geographical distribution of Infusoria (a
general grouping which includes diatoms) in North Africa and West Asia.
This paper was followed by several others on the distribution of Iitfusoria
in various parts of the world. Other workers such as Greville, Kutzing,
W. Smith, Grunow, Van Huerck and Cleve continued to explore this
field of interest. They recorded not only the geographical distribution
of diatoms but also described the conditions in which diatoms were found.
Cleve (1894) pointed out the importance of a knowledge of the habitats
and geographical distribution of diatoms to geological research. Consider'
able information has been obtained by studies of fossil diatoms as to the
extent of the invasion of the sea and the effect of glaciation on the temp-
erature of fresh and marine waters. Much of the knowledge of thepaleo-
geography of Scandinavia has been elucidated in studies of fossil diatoms
by Cleve (1899). Cleve-Euler (1940, 1944), Hustedt (1939) and Molder
(1943).
In North America the extent of the effect of glaciation on fresh waters
has been set forth by studies of diatoms (Hanna, 1933; Patrick, 1946).
The succession of changes occurring in lake development has also been
determined by diatom studies of Patrick (1936, 1943, 1946 , 1954),
Pennington (1947) and Ross (1950). It is because many species of diatoms
have their best development in water with certain specific chemical
characteristics that such correlations are possible.
One of the most important works summarizing what was known as
to the ecology of diatoms was published by Kolbe (1927). In this work
he clearly sets forth a system for classifying diatoms as to their
tolerance to various chloride concentrations in water. Krasske (1927),
Hustedt (1953), Legler and Krasske (1940) and Petersen (1943) have also
contributed to our understanding of the chloride tolerance of Hiatoms.
-71-
-------
The knowledge of the relationship of the occurrence of given species
of diatoms to other chemical characteristics of water, such as pH, iron,
nitrates, phosphates, and silicon, has developed greatly during the last
25 years. From time to time this kind of information has been brought
together and published by such workers as Hustedt (1938 - 39), Schroeder
(1939), Patrick (1948) and Fjerdingstad (1950),
Only very recently has the structure of the diatom population been
correlated with the presence of sanitary and industrial wastes . Kolkwitz
and Marsson (1908) set forth a system classifying many species oftdiatoms
as to their ability to withstand varying degrees of pollution from sanitary
sources. Fjerdingstad (1950) published a very good summary of the more
important literature on the effect of varying amounts of pollutants on the
occurrence of diatoms. Budde (1930b) found out that the association of
species and the relative sizes of populations of species produce the best
picture of the effects of waste on the diatom flora. Although many of the
studies concerned with the effect of wastes on the aquatic life of a stream
have been based on plankton, Jurgensen (1935), Nowak (1940), Butcher
(1933, 1940), Patrick (1948) and others have pointed out that it is the
attached forms, or those organisms which grow and reproduce in a given
area, that give the most reliable indication as to whether the environment of
an area is suitable for the support of aquatic life .
Diatoms may float or be carried into habitats where they may survive
far a period of several days without dying, although the quality of water
is unsuitable for growth and reproduction. This is one of the reasons why
diatoms, which in natural or unpolluted rivers indicate so well changes in
the environment, may under conditions set forth abave persist and thus
not appear to indicate the character of the water in which they are found.
Another error which has occurred in the study of diatoms has been
pointed out by Fjerdingstad (1950). Workers often fail to examine their
collections to see if the diatoms found are living and in good condition.
Since diatoms have cell walls of silicon, which persist after they are dead,
they may be included as living in a given area unless the above precaution
is taken. In our studies of diatoms taken from various rivers in the
United States, large numbers of dead frustules of species not living in an
area are often found intermixed in the liviiig collections.
Another cause of error in the handling of collections is contamination.
Dirty pipettes, beakers and collecting jars may well result in the transfer
Of species of diatoms from one collection to another and thus produce a
false picture of the structure of the diatom flora being studied.
-72-
-------
The methods used in collecting diatoms and of studying the
slides must be the same if one wishes to compare the diatom flora
of various areas. Because these methods are not standardized
among diatomists it is often very difficult to compare results.
Such considerations are particularly important if one is comparing
the number of species and the sizes of the populations.
Another consideration which is often overlooked in discussing
indicators of polluted or of deleterious conditions is the number of
variable factors and the combination of such factors which may produce
the deleterious effects. As pointed out by Fjerdingstad (1950) the
classification assigned by Kolkwitz and Mars son (1908) to certain diatom
species often does not hold. This undoubtedly is due to several factors
but certainly one of the most important is the fact that Kolkwitz and
Mars son's system is based on the reaction of organisms to sanitary
wastes or wastes with a heavy organic load.
In this system the greatest degree of pollution is characterized by
low oxygen content, high bacterial counts, high biochemical oxygen
demand and a heavy organic load. However, when one is considering
toxic wastes, a stream may have a high dissolved oxygen content, low
biochemical oxygen demand and low bacterial counts and yet be inimical
to aquatic life because of the presence of toxic substances. Likewise
high temperatures may be lethal and only be associated with a lowering
of the dissolved oxygen. Whereas organic wastes, toxic wastes and
high temperatures in excessive amounts will kill most organisms in
sublethal or threshold concentrations they will affect various species
in different ways. In most rivers which we have studied in the United
States, pollution is rarely of a single type, but rather is a combination
of toxic substances and organic wastes often accompanied by high
temperatures.
Fjerdingstad (1950) and others have emphasized that various aspects
of the diatom flora must be considered, such as the changes in numbers
of species, numbers of individuals and kinds of species. As a result
of over 100 analyses of diatom floras from natural areas of rivers in
eastern and southern United States, our laboratory ha's found that tlie numbe
of species making up the diatom flora are quite similar. By natural
areas are meant areas which so far as we could ascertain fr6m state
and federal agencies as well as from information concerning chemical
and bacteriological analyses were not adversely effected by effluents
entering the river. Indiscriminately selected results from such studies
showed the following numbers of species to be present: Marsh Creek,
Pa. - 46; Tionesta Creek, Pa. - 42; North Fork of the Holston, Va. -
52, Savannah River, South Carolina and Georgia - 80, 69; Flint River,
Ga. - 40; Escambia River, Fla. - 55, 54; Sabine River, Texas - 46,
55; Guadalupe River, Texas - 42; Neches River, Texas - 65.
-73-
-------
Part of the variation shown here in the numbers of species present
in the various studies is due to the fact that the studies on Marsh Creek,
Tionesta Creek, Flint River, Sabine River and Guadalupe River were made
in a somewhat different manner than those on the other creeks and rivers.
When one considers the kinds of species a very different picture is
presented. In a study of natural river areas soon to be published, 354 species
from ten rivers were considered. Of this number only one was found in all
ten rivers. Two hundred and two were found in only one river; 74 in two*;
30 in three; 20 in four; 9 in five; 5 in six; 9 in seven; 3 in eight and 1 in nine.
This clearly shows the difficulty of using single species as indicators. On
the other hand, if one considers a group of species as indicating a condition
a reliable methodology can be developed. For instance, most of the species
of Eunotia and all of the North American species of Actinella are found in soft,
usually somewhat acid waters. The presence of an association of any of the
species of these genera will indicate these conditions. Likewise, the presence
of well developed populations of Synedra affinis or S. pulchella or Navicula
pygmea will indicate the presence of brackish water. As a general rule,
species which form the largest populations should be selected as indicators.
In other words diatoms most truly indicate those physiological conditions
which enable them to multiply most rapidly over a period of time.
An analysis of the histograms given by Patrick (1949) shows that the
diatom flora responds to the severe effects of deleterious effluents in a
manner similar to that of fish and insects. Approximately 200 analyses of
sections of rivers and estuaries which have been made by the Limnology
Department of the Academy of Natural Sciences of Philadelphia subsequently
support these conclusions.
Diatoms are in some cases more sensitive to small changes in the
chemistry of the water than are some of the larger forms, as was pointed
out by Liebmann according to Fjerdingstad (1950). However, the response
of diatoms to lethal concentrations are usually similar to those of fish.
Analyses of results of 50 bioassay tests run on wastes of chemical,
steel, gas and electric industries show that the concentrations which prd-
duced 50 per cent reduction in growth of diatoms and 50 per cent kill in
fish were very similar (Table 1). This work was carried out by Dr. John
Cairns, Jr. and Dr. Arthur Scheier. In 7 5 per cent of the cases one
concentration was never more than twice that of the other, and in most
cases there was very little difference. Since the concentration recommended
as biologically safe is less than a third of the above concentration, it is
evident tha^aresults from either test will bring about very similar recommen-
dations "Tor safe discharge of wastes. In only eight per cent of the cases
was the diatom found to be less Sensitive than the fish. These results clearly shov*
-74-
-------
Table 1
Comparison of effects of industrial wastes on fish and
diatoms (Figures in column headed "Diatom" represent
concentration, in per cent, of effluent causing 50 per cent
reduction in division rate in five days; those under Fish
show concentrations causing death to 50 per cent of the
fish in 24 hours and in 48 hours. )
Fish
Effluent
Diatom
24 hours
48 hours
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
5. 3
2. 78
3. 65
0. 001
1. 16
2. 3
no reduction
49. 3
2. 34
0. 78
2. 15
18. 7
4. 4
32. 8
65. 0
35. 0
44. 5
no reduction
0. 74
no reduction
no reduction
56. 0
18.0
no reduction
no reduction
no reduction
98. 0
no reduction
no reduction
no reduction
10
2. 8
3. 2
0. 18
3. 25
7. 6
70. 0
no deaths
10. 0
0. 57
3. 2
26. 5
75. 0
62. 0
no deaths
no deaths
1.6
12. 7
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
9
2. 8
2. 7
0. 155
2.87
6. 0
60. 0
90. 0
9. 1
0. 52
3. 2
21. 7
75. 0
52. 0
no deaths
no deaths
1.6
9.0
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
"No death" or
was not deleterious
"No reduction" means that 100% concentration of the effluent
-.75-
-------
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
Table 1 (Continued)
Fish
24 hours
48 hours
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
no deaths
6. 9
20. 5
0. 44
42. 0
no deaths
29. 5
no deaths
Diatom
no reduction
no reduction
no reduction
56. 0
18. 0
no reduction
no reduction
no reduction
98. 0
no reduction
no reduction
60. 0
no reduction
6. 0
10. 0
0. 77
16. 2
no reduction
15. 0
no reduction
-76-
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value of diatoms as indicators of conditions essential for or inimical
to fish life.
Results of work done by the Limnology Department under grants
from the U. S. Public Health Service indicate that when pure chemicals
are tested singly, responses of fish and diatoms are more varied.
More study is needed before an explanation can be given for the
more variable responses to pure chemicals than to industrial effluents.
The differences were not due to procedure since the same methods
were used in both types of tests.
From the many studies that have been made it is apparent that
diatoms can be used as a group to indicate the ability of a water to
support aquatic life. Collections must be prepared correctly and
the studies based only on specimens living at the time collections are
made. Furthermore, collections should be taken from populations which
are attached or definitely living in the habitat and not from plankton
forms.
Diatoms are a desirable group to use for indicating stream conditions
for several reasons:
1. They need no special treatment for preservation because the
cell wall, on which the identification is based, is composed of silica.
2. The diatom flora of a normal stream is made up of a great
many species and a great many specimens. Thus the group lends itself
to statistical treatment.
3. Diatoms vary greatly as to their sensitivity to chemical and
physical conditions of water. Some species are able to tolerate a
wide variety of environments. Therefore, some diatoms can be found
in any aquatic habitat inhabitated by plants and animals (excluding
bacteria). However, each of these species has a range of conditions
in which it achieves best development. Other species of diatoms have
a very narrow range of tolerance. Thus we have in the diatoms
enough different kinds of species so that they can be used as in indication
group in most of the possible types of aquatic environments found in
rivers and estuaries.
4. A considerable amount of information is already available as
to the type of environment in which many species are found.
In order to use any group of organisms as indicators one must
have a method of collecting and studying them which will be comparable
for different types of water. A suitable methodology for diatoms has been
set forth by Patrick, Hohn and Wallace (1954) by the use of the
Catherwood Diatometer.
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The Diatometer (Fig. 1) is an instrument which floats in the
water and provides a substrate, ordinary glass slides, which
seems to be non-selective for diatom growth. These slides are
left in the water for a long enough period of time to allow a consider-
able amount of growth to develop on the slide. In many cases this
length of time has been about two weeks. From the studies we have
made, it appears that dead diatoms do not remain attached, but
slough off the slides. Studies based on these slides thus include
only live diatoms which are actually living in a given area.
When the slides are prepared for study the diatoms are scraped
off and cleaned by the acid method. A small aliquot which is
representative of the material is then placed on a slide and mounted in
Hyrax.
In these studies all species observed are identified and the number
of indivduals of each is recorded. Enough specimens have to be identi-
fied in order to construct a truncated normal curve according to the
method set forth by Preston (1948). If results are to be comparable
the modes of the curves should be in the same interval. Therefore,
varying numbers of specimens may be counted. This study usually
takes 3 to 4 days to complete.
In such rivers as the Savannah where little change occurs, the
structure of the curve remains very similar from season to season and
from year to year. However, any serious change in the quality of
water is readily seen by a change in the structure of the curve (Figs.
2, 3, 4). In interpreting these curves one must take into consideration
the height and position of the mode, the dispersion factor, the number
of species found, the total theoretical population, the kinds of species
and the number of specimens used in constructing the curve.
It can be seen that this method avoids many of the pitfalls which
have been encountered by trying to use diatoms as indicators of pollutioi
It provides a uniform method for collecting and studying diatoms. Thus
the results are comparable. This method is based on the study of the
live species in a given area which are growing and dividing. It is mainly
concerned with the number of species and number of individuals of each
of the species. It also considers the kinds of species. Thus its basis
is a shift in pattern of the whole flora rather than the behavior of a
few indicator species.
Of course, it must be remembered that any conclusion based on onl';
one group of organisms or kind of analyses should only be considered as
giving an indication of conditions. This applies not only to biological
analyses but physical and chemical analyses as well. If one wants a
complete picture of river or estuary conditions then all groups of
organisms as well as chemical and physical analyses must be included.
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Fig. I
INDIVIDUALS PER SPECIES
Graph of a Diatom Population from a River Showing
Mild Effects of Pollution.
Fig. 3
INDIVIDUALS PER SPECIES
Graph of the Diatom Population from a River Not
Adversely Affected by Pollution.
Fig. 2
INDIVIDUALS PER SPECIES
Graph of a Diatom Population from a River Showing
Severe Effects of Pollution.
Fig. 4
-------
To date this type of study of diatoms has been found to indicate
reliably the quality of water as to its ability to support aquatic life
Although in most cases it indicates the condition not only of the water
but also of the river bed, two exceptions have been found. These
were areas in which the effluents entering a river had recently been
greatly curtailed but the condition of the bottom was such that sessile
or burrowing forms could not live in it. Thus the quality of water,
as verified by bioassay tests on fish and invertebrates, was good
even though the bottom was poor.
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References
Budde, H. 1930a. Die Algenflora der Ruhr. Arch. f. Hydrobiol.
21: 559-648.
1930b. Die mesohaloben und halophilen Diatomeen der
Lippe in Westfalen. Ber. Deutsch. Bot. Ges. 48:415-419.
Butcher, R. W. 1932. Studies in the ecology of rivers. II. The
microflora of rivers with special reference to the algae on the
riverbed. Ann. Bot. 46:813-861.
1940. Studies in the ecology of rivers. IV. Observations
on the growth and distribution of sessile algae in the River Hull,
Yorkshire, Jour. Ecology 28:210-223.
Cleve, P. T. 1894. Synopsis of naviculoid diatoms. K, Sv. Vet. Akad,
Handl. , 2nd ser. , 26(2):12.
1899. Postglaciala bildningarnas klassifikation pa grund
af deras follisa diatomaceer. Sv. Geol. Und. , Ser. C. No. 180:59-
61.
Cleve-Euler, Astrid. 1940. Das letztinterglaziale Baltikum und die
Diatomeenanalyse, Beih. Bot. Zentralbl. , Abt. B, 60{3):287-334.
,,1944. ,pie Diatomeen als quartargeologische Indikatoren.
Geol. Foren. Forhandl. 66(3):383-410.
Ehrenberg, C. G. 1829- Die geographische Verbreitung der
Infusionsthierchen in Nord-Afrika und West-Asien, beobachtet
auf Hemprich und Ehrenbergs Reisen. Abh. K. Akad. Wiss,
Berlin, 1829:1-20.
Fjerdingstad, E. 1950. The microflora of the River M/^lleaa.
Folia Limnol. Scand. No. 5, 123 pp.
Hanna, G. Dallas. 1933. Diatoms of the Florida peat deposits.
Ann. Rept. Florida State Geol. Surv. 23/24(1930/l932):68-l 19.
Hustedt, Fr. 1937-1939. Systematische und okologische Unter-
suchungen uber die Diatomeen-Flora von Java, Bali und Sumatra.
Arch. f. Hydrobiol. Suppl. 15:131-177, 187-295, 393-506, 638-790;
Suppl. 16:1-155, 274-394.
1953. Die Systematik der Diatomeen in ihren Beziehungen
" zur Geologie und Okologie hebst einer Revision des Halobien-
Systems. Sv. Bot. Tidsk. 47(4):509-519.
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Jurgensen, Charlotte. 1935. Die Mainalgen bei Wurzburg. Arch,
f. Hydrobiol. 28:361-414.
Kolbe, R. W. 1927. Zue Okologie, Morphologie und Systematik der
Brackwasser-Diatomeen. Pflanzenf. 7:1-146.
1932. Grundlinien einer allgemeinen Okologie der
Diatomeen. Ergebnisse d. Biol. 8:221-348.
1954. Einige Bemerkungen zu drei Aufsatzen von Fr.
Eultedt. Bot. Notiser l954(3):217-229.
Kolkwitz, R. , und M. Marsson. 1908. Okologie der pflanzlichen
Saprobien. Ber. Deutsch. Bot. Ges. 26:505-519.
Krasske, G. 1927. Diatomeen deutscher Solquellen und Gradierwerke.
I. Arch f. Hydrobiol. 18:252-272.
1939- Diatomeen deutscher Solquellen und Gradierwerke.
ill. Beih. Bot. Zentralbl. , Abt. B, 59(3):413-436.
Legler, Fr. und G. Krasske. 1940. Diatomeen aus dem Vansee .
(Armenien). Beitrage Zur Okologie der Brachwasserdiatomeen.
I. Beih. Bot. Zentralbl., Abt. B, 60(3);335-345.
Molder, K. 1943. Rezente Diatomeen in Finnland als Grundlage
quartar-geologischer Untersuchungen. Geol. d. Meere u.
Binnengew. 6(2): 148-240.
Nowak. W. 1940. Uber die Verunreinigung eines kleinen Flusses in
Mahren durch Abwasser von Weissgerberein, Leder-,
Leimfabriken und anderen Betriebeji. Arch. f. Hydrobiol. 36:386-
423.
Patrick, Ruth. 1936. Some diatoms of Great Salt Lake. Bull.
Torrey Bot. Club 63(3): 157-166,
1943. The diatoms of Linsley Pond, Connecticut. Proc.
Acad: Nat. Sci. Phila. 95:53-110
1946. Diatoms from Patschke Bog, Texas. Not. Nat. ,
Acad. Nat. Sci. Phila. No. 170, 7 pp.
1948. Factors effecting the distribution of diatoms. Bot.
Kev7 14(8):473-524.
1949. A proposed biological measure of stream conditions,
based on a survey of the Conestoga Basin, Lancaster County, Penn-
sylvania. Proc. Acad. Nat. Sci. Phila. 101:277-341.
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1954. The diatom flora of Bethany Bog. Jour.
Protozool. 1:34-37.
Patrick, Ruth, M. H. Hohn, and J. H. Wallace. 1954. A new method
for determining the pattern of the diatom flora. Not. Nat. ,
Acad. Nat. Sci. Phila. No. 259. 12 pp.
Pennington, W. 1947. Studies of the post-glacial history of British
vegetation. VII. Lake sediments: Pollen diagrams from the
bottom deposits of the north basin of Windermere. Phil. Trans.
R. Soc. London, Ser. B. , No. 596, 233:137-175.
Petersen, J. Boye (Boye-Petersen). 1943. Some halobien spectra
(diatoms). Biol. Medd. Dan. Vid. Selsk. 17(9):l-95.
Preston, F. W. 1948, The commonness and rarity of species.
Ecology 29:115-116.
Ross, R. 1950. Report on diatom flora from Hawks Tor, Cornwall.
Phil. Trans. R. Soc. London, Ser. B, No. 615, 234:461-464.
(Appendix in: Studies in the post-glacial history of British
vegetation. XI. Late-glacial deposits in Cornwall, by Ann P.
Conolly, H. Godwin, and Eleanor M. Megaw. )
Schroeder, H. 1939- Die Algenflora der Mulde. Ein Beitrag zur
Biologie saprober Flusse. Pflanzenf. 21:1-88.
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USE AND VALUE OF FUNGI AS BIOLOGICAL INDICATORS OF POLLUTION
Wm. Bridge Cooke
Robert A. Taft Sanitary Engineering Center
U.S. Public Health Service
Cincinnati, Ohio
A person who has spent more time studying North American forest
than the streams which flow through them thinks of indicators in terms of
trees. The presence of subalpine fir in Paradise Valley on Mount Rainier
indicates that one is in the spurce-fir zone near the upper limit of trees on
that type of mountain in that geographical location; the presence of Joshua
trees in southeastern California indicates that one is in one phase of the
Mojave Desert; the presence of sycamores and American elms in a shallow
valley indicates that one is near a stream in the vast eastern deciduous or
mixed mesophytic forest.
But these trees all indicate that one is on relatively undisturbed land.
Our immediate concern is based on whether or not fungi can be used as indi-
cators of pollution. It has been said (Suter and Moore, 1922) that the Sapro-
legniaceae can be used as indicators of pollution. It has been said (Suter
and Moore, 1922; Butcher, 1932; Cooke, 1954) that Leptomitus lacteus is
an indicator of pollution. The filamentous bacterium, Sphaerotilus natans,
in its several forms, and the stalked protozoa including Carchesium have bee
referred to as fungi which indicate pollution (Butcher, 1932; Cooke, 1954).
That little interest has been shown in analysing the components of "sewage
fungus" may be seen in Wilson's paper on the microbiota of sewage published
m 1944 .. Here, "fungi indet." is simply one category among the many types
of organisms listed according to genus . The number and types of fungi
found in polluted water and sewage to date have only indicated that certain
strains of a number of common soil fungi have become adapted to or are
able to tolerate this different habitat.
Materials from several types of fungus studies will be drawn on in an
attempt to determine something of the relationship between fungi and their
habitats with special emphasis on pollution.
From the beginning of life, various members of the populations of the
world have been leaving their bodies or the remains of their metabolic pro-
cesses on the land or in the water to continue the process of decay so that
:he organic materials of which they were composed may be used and reused.
One type of intermediate organism in the waters of the world is the group
:ommonly called "sewage fungus". These organisms developed a specialized
ype of metabolism in which certain forms of carbon and nitrogen compounds
:an be utilized but not others. As the numbers of man increased, and as
lis physical requirements increased, the waters of the world have increasingly
:arried the waste products resulting from this development, and organisms
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adapted to the special nutrients found therein have thrived and multiplied.
In this way, we may suppose, such organisms as Sphaerotilus and Leptomitus
have developed, multiplied;, and filled their special ecological niches"! Lep-
tomitus has developed to the point that it uses fatty acids rather than sugars ,
and amino acids rather than ammonia (Cantino, 1955).
There are at least three sources of pollution by which food materials
usable by fungi are placed in streams. The first may be considered natural
pollution. Pieces of decaying vegetable or animal materials may inadvertently
reach streams through run-off, passive falling into the water, or other means.
We are not concerned with this type of pollution. Many of the true water
molds or aquatic fungi - Saprolegniales and related fungi - are found only
under these conditions. They require something to attach themselves to
during their vegetative phases . Such fragments of materials may serve as
food as well as anchoring places for these water molds. Other water molds,
belonging to the Chytridiales, do not always live freely in the water but
may be parasitic upon various species of algae and in various kinds of pollen
which float on the water usually in the spring. When their sources of nutri-
tion become exhausted, these fungi produce resting spores and cells ("gemmae")
The second source of pollution is fecal material and other materials
such as ground garbage added to the stream in domestic sewage. Such
materials are high in organic content and may furnish large amounts of
foods to fungi that have become adapted in one way or another to life in the
water or in the stream bed. Under restricted conditions , with certain
carbohydrate, nitrogenous, and as yet undetermined additives, and under
certain conditions of substrate and bottom materials resulting in special
pH and mineral content conditions, certain Saprolegniaceae, Leptomitus
lacteus, Sphaerotilus natans, and various soil fungi, such as iFusarium
aquaeductuum, Geotrichum candidum , Penicillium lilacinum , and others ,
may thrive. Under these conditions, such organisms appearing in large
numbers will indicate the presence of polluting substances, substances which
make the waters of the stream unfit for human and most industrial uses.
The third source of pollution is various types of wastes from industry.
Different types of industrial pollutants can produce varying conditions of
growth in the vicinity of the point of discharge of the pollutant. Such pollu-
tants will become diluted downstream. It is possible that, at the immediate
outfall, oxygen supplies may be too poor and certain substances may be too
toxic to support growth, but as oxygen increases and the toxic substances
become diluted by increased flow, precipitation, or other factors, the toxic
effect will be weakened and growth of various fungi and fungus-like organisms
may be encouraged or enhanced. When the pollutant is organic, Leptomitus
lacteus and the filamentous bacterium Sphaerotilus natans may thrive under
such conditions; and at least Sphaerotilus will present several growth habits,
mistaken as species, genera, or varieties in the literature, as the concentra-
ted waste becomes more dilute. At Lytle Creek (Cooke, 1954c),
a number of species of fungi have been found in isolations from
water, sediment from polls and riffles, and apparently septic bank
soil, at the outfall of primary settled sewage .
-85
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Both Leptomitus and Sphaerotilus have been cultured in the laboratory
with little difficulty after the proper combination of nutrients has been
determined. Such combinations indicate only that these organisms have
special requirements for sugars and compounds containing nitrogen. In
nature such substances are present largely under conditions of pollution by
organic wastes from the home or factory; in the wilderness , such substances
may be present following death and during decay of plants or animals. It
is quite possible that these organisms may be common around watering holes
and other places where animals have congregated, where their remains or
droppings may have produced local natural pollution.
Another type of fungus may be thought to be more nearly indicative of
pollution, especially of the domestic type. In the forest we find these co-
Prophilous fungi associated with dung of wild animals, rarely of man. In
many cases these fungi are restricted to specific types of dung: Mucor
ramanianus is found on frog droppings; Pilobolus species are usually found on
Horse droppings . Other types of these fungi are found on the dung of any
animal that uses plants as food. The spores of these fungi are so produced
and discharged that they become attached to or glued to leavers of plants
®ed as food by grazing or browsing animals. After the spore has passed
through the alimentary canal of an animal, it is able to germinate, use
;he dung as food, and fruit quickly after deposit of the dung. Ephemeral
*iushro-oms like the inky cap and the common mushroom are edible, but
since man.qooks the mushrooms he eats, their spores are killed before
¦ngestion. Few if any coprophilous fungi are found associated with human
sees, especially where this material accumulates as night soil or is carried
iway in sewer's to rivers or sewage treatment plants. While Sordaria humana
>ccurs on others £ypes of dung, the specific name indicates that man also can
ngest such fungjas^ spores and that under certain conditions these fungi can be
'ecovered from 'h^r^an wastes .
Three types, of $ingi which may be placed in this physiological class
iave been found in sewage polluted water and on trickling filters . A species
f Pilobolus (HarveyV 1952) was found at one station on Lytle Creek by use of
emp seed as an i sola ting vme dium. It was found again on the Glendale,
>hio, trickling filter. Intensive biochemical nutrition research (Hesseltine,
t al0, 195 3) has shown that this fungus can be propagated in the laboratory
'ithout ingestion and alimentation by an animal,but the technique has not
een tried at Cincinnati. Ascodesmis microscopica was found only twice
Seaver, 1928) in the wor,ld prior to 1955. One of these growths was in
lurope on tiger dung, the other at the Bronx Park Zoo on the dung of a
accoon dog. In 1955 the species was found on a trickling filter in the Dayton,
>hio, sewage treatment plant. It requires no special techniques or media
ut will grow on any medium used in culturing the so-called sugar fungi.
In 1953, Hesseltine, studying the fungi of trickling filters at Pearl
iver, New York, found a fungus (Subbaromyces splendena) which in pure
alture grows only on lima bean agar. This has since been found on
-------
trickling filters at Dayton, Ohio, and Pullman, Washington (Becker and
Shaw, 1955). It is possible that this fungus may grow in other habitats
with similar environments, such as on stones of streams polluted with domes-
tic wastes and on other trickling filter beds.
An organism can become adapted to one set of environmental conditions
or another. In some cases the adaptation may be completes so that the
organism cannot tolerate any other conditions, but more often various de-
grees of adaptation may be attained by an organism. Moser (1949), study-
ing the adaptation of certain fungi, mosses, and seed plants to areas in the
tyrolean forests which had been burned by fires ranging from large forest
fires to small camp fires s developed the following set of terms to describe
the degree of adaptation of the organisms to burned areas: anthrocobiont
species were found only on burned soils and are not known to occur elsewhere;
anthracophilous species are found more commonly on burned areas; anthra-
coxenous species occur more commonly on unburned areas but will tolerate
burned areas; while anthracophobous species will not grow on burned areas.
This set of suffixes can be used to describe other habitat requirements of
the same or different species. Since "copro-" has been used commonly
in the combination "coprophilous11 for a fungus growing on dung, it might
be confusing to use in the terms we want. The Greek work "lyma" means
"filth" . Then, lymabiont species will grow only on or in the presence of fecal
materials; lymaphilous species will grow commonly on such material but
will also grow on other materials; lymaxenous species will grow commonly
on other materials but will tolerate fecal matter; while lymaphobous species
will not grow on or in the presence of fecal material. In this sense, a co-
prophilous species may be either a lymabiont or a lymaphile. A lymaxene
could be an organism which on occasion will colonize the substrate material,
while a lymaphobe will not grow on such materials. The emphasis here is
on nutrient requirements or tolerances, rather than on habitat types or
tolerances as is the emphasis in the Kolkwitz (1950) system.
To equate this set of terms with those of Kolkwitz, the following parallels
are suggested. Polysaprobes would include both lymabionts and lymaphiles.
For use in connection with fungi, polysaprobe would be confusing since it
implies the ability to live saprobically on any substrate. Alpha (or strong)
mesosaprobes would include lymaphiles primarily, but lymabionts and
lymaxenes could be expected to be found. Beta (or weak) mesosaprobes
would include lymaphiles, but lymaxenes could be found more frequently than
in the preceding categories. Oligosaprobes would include the lymaphobes
and probably some lymaxenes and lymaphiles.
Indicators of fecal pollution will thus be found in the first two cate-
gories, lymabionts and lymaphiles, rather than restricted to the first cate-
gory. Coprophilous species connote only those species of fungi or other
organisms growing on dung at the time of observation, so the term is not
useful for our purposes. Of the lymaphilous species, one must be careful
that the habitat is described in more than one way. For instance, the fungus
Fusarium aquaeductuum was first described from wooden water pipes which
under certain conditions could be plugged with its massive growth. At present,
wooden pipes are used infrequently, pure water systems have been rarely
-87-
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checked for the fungus, and it is abundant on trickling filters throughout
the country as well as in polluted streams .
In intensively cultivated fields there is strong competition for nutrient
materials among the various species, varieties, strains, and clones of fungi
present. Such competition has given rise to species that have become adapted
to the utilization of special nutrients (Garrett, 1956). A strictly saprobic
species will attack almost any dead organic material. Certain species are
able to attack youngseedlings, produce seedling death, and then use the dead
tissues as nutrient sources for primary colonization of new substrata. Other
species can penetrate the roots of older plants, grow through their vascular
tissues j, and produce the type of disease referred to as vascular wilts.
Such fungi in the soil may be considered ecological obligate parasites, for
although they can be readily cultured on any agar medium on which sugar
fungi develop, in nature they cannot compete with those fungi that colonize
dead tissue, but must create their own dead tissue by killing the host. As
the host plant dies, the fungus can utilize its tissues as food until forced into
retirement by competition from other fungi or depletion of nutrient materials .
These vascular wilt fungi belong to the genera Fusarium and Verticillium,
among others. Three species of the wilt-producing fungi, F. oxysporum,
F. roseum, and F. solani, have been isolated occasionally from sewage and
polluted water. "The source of these fungi in the samples that have been studied
is unknown, but the species are widely distributed agricultural pests. Nothing
is known of their activities away from the soils in which they are of greatest
importance as plant disease producing fungi, except that they are capable
of causing deterioration of cellulose. In preliminary experiments in dilution
water it was apparent that a strain of F. oxysporum is capable of using
hydrocarbons as its sole source of carbon"! "fhat these organisms are isolated
occasionally or even commonly from sewage and polluted water indicates
that they are capable of competing with other fungi and other organisms for
whatever nutrients are offered. In contrast to the soil, where only occasionally
does dead organic matter become available to the fungi of a specific unit of
soil, or where only occasionally does a plant root become available for colon-
ization even in a field where the crop is relatively densely planted, polluted
Water presents to the fungus a continuous source of nutrient supply. Such
a supply makes it unnecessary for the fungus to assume a resting phase upon
the depletion of one source of nutrient while waiting for a new source.
Nutrition requirements of organisms found in industrial types of pollution
are more difficult to define. Albritton (1955) is very general on the point
of fungus nutrient requirements. Fecal material is at a minimum in such
wastes, while sugars and possibly organic nitrogen sources are of greater
importance. Here a somewhat different type of organism will be ascendant,
although many of the same species will occur commonly in both types of
pollution.. Again, where a waste contains metallic ions usually inhibitory
to the growth of organisms, certain species may become adapted to life in
the presence of those substances. For instance, at least one species of
Penicillium, P. ochrochloron,can tolerate high concentrations of copper,
another, manganese. Some fungus contaminants can be found commonly in
-------
acids thought to be toxic to any growth. In general, such fungi fall into the
broad classification recently proposed: "sugar fungi". These fungi are able
to utilize simple sugars but in many cases are not able to develop an enzyme
system for the degradation of more complex carbohydrates, or other carbon
sources.
A further difficulty with the definition of a pollution indicator among the
fungi is the inability of the investigator to identify most of the fungus growth
in any one sample on sight or on preliminary microscopic examination.
Most soil fungi growing in an aqueous habitat do not produce spores but
form an extensive mycelial mat. The mat may be formed by the interweaving
of a number of mycelia of many species . The orange color produced by
Fusarium aquaeductuum is a result of large numbers of spores piled together
or produced concurrently. The white color produced by Geotrichum candidum
on surface films of trickling filters is quickly masked by algae or other fungi,
or the filamentous bacterium Sphaerotilus natans which produces a grey
color; or, it may be confused with the occasionally produced white mat of
Leptomitus lacteus when that is able to appear. In some cases, species of
Penicillium may be observed in fruiting condition, but it is rare that a typical
fruiting structure will be produced; the phialides producing the spores may
be found singly on the hypha or in atypical clusters, rather than clustered
in the typical penicillate brush at the tip of the spore-producing hypha. It
is virtually impossible to study the several genera of the white yeasts,
the red yeast, and the black yeast by direct observation of scrapings from
trickling filter stone or stones in creeks or rivers subjected to continual
pollution loads. Such studies must be made by using cultural techniques and
studying the colonies that develop directly on properly prepared dilution plates
or later in pure culture.
Techniques used most successfully so far in the isolation of fungi from
polluted water and sewage have been described by Cooke (1954b). For
isolation of aquatic fungi the technique described by Harvey (1952) is adequate.
For help in studying the fungi that have been isolated, several books are
available. The various media used in the study and identification of yeasts
are described by Lodder and Kreger-van Rij (1952). The genera Aspergillus
and Penicillium are best studied by techniques defined respectively by Thorn
and llaper (1945) and by Raper and Thom (1949). When species obtained by
baiting or using hemp seed according to Harvey's method are obtained, they
can be studied with the help of Coker (1923) or Coker and Matthews (1937).
If species of Pythium appear following use of this technique, Middleton
(1943) is useful"! Parasites of plankton, pollen grains, and similar substrata
can be studied with the help of Sparrow (1943). Of other fungi, that have
not been studied consistently on one medium or do not belong in the categories
mentioned above, some groups cannot be satisfactorily studied at present.
These include primitive Ascomycetes, mycelial Basidiomycetes, sterile
mycelia, members of the Sphaeropsidales, and such genera as Chaetomium.
For none of these groups are there adequate taxonomic treatments, and
cultures must be sent to specialists. Of the Sphaeropsidales, most known
specie's are studied with reference to a specific host plant. Cultures from
water, sewage, or soil, therefore, cannot be identified satisfactorily without
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««-accuing tne potential tiost plant population of the region. To aid in the
identification of a genus of fungus , the worker can consult Clements and
Shear (19 32), whose keys are lengthy and cumbersome, or he can study
Barnett (1955), who has illustrated nearly one-third of the genera including
those that are more common For more specific information, Gilman's
(1945) compilation is useful,, and Smith (1954) gives considerable help with
the commer species found in industry and the laboratory. Finally, Hughes
(195 3) points toward newer concepts in developing a workable system of
classification of the so-called mold fungi,
A technique for obtaining samples of growth from trickling filters,
modified from similar techniques whose origin dates back to 1905, has
been developed in whichs rather than scraping ths stone, glass slides are
placed in mounts in contact with the growth on the stone and continually
irrigated by the settled sewage spray applied to the filter beds , Growth on
the slides can be removed by scraping with a rubber policeman, the algae
counted, the protozoans studied, and the fungi and bacteria plated. If the
material to be studied for development of fungi is broken up in a Waring
Blendor, an approximation of the importance of each fungal species can be
obtained by counting the number of colonies of each species or of all species .
Using this technique, it appears that 10 to 15 species of soil fungi form
the largest part of the fungal portion of the slimes on trickling filters.
Annotated Bibliography
Albritton, E G.s 1955, Standard Values in Nutrition and Metabolism.
xiji, 380, Philadelphia, W, B„ Saunders Co.
A tabular presentation of the nutrient requirements of the prin-
cipal groups of organisms, of certain specific groups and of certain species.
Tables also present food and feed values of certain species of organisms,
and metabolic products of various organisms used in industry.
Barnett, H, L., 1955. Illustrated Genera of Imperfect Fungi, i, 218,
Minneapolis, Burgess Publishing Co.
The commoner genera of mold fungi and plant pathogens are illus-
trated with line drawings . Habitat descriptions and references are given
for each species illustrated.
Becker, J. G, and C. G, Shaw, 1955. Fungi in domestic sewage treatment
plants. Applied Microbiology 3: 173-180
A study of fungi found in the sewage treatment plants at Pullman,
Washington, and Moscow, Idaho, based on studies of effluents from various
process points in the two plants which treat domestic sewage.
butcher, R. W,, 1932, Contribution to our knowledge of sewage fungus.
Trans . Brit. Myc. Soc . I7j 112-124
A systematic study of the various organisms which are associated
With polluted water and sewage treatment plants and which have been termed
"sewage fungus".,
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Cantino, E. C., 1955. Physiology and phylogeny in the water molds - A
reevaluation. Quart. Rev. Biol. 30: 138-149.
A study based on the writer's personal experience with some of
the groups of organisms listed as well as on a thorough knowledge of the
literatures in which an attempt is made to base a phylogeny of the water
molds on what is known of their physiology.
Clements , F . E . and C. L Shear , 1931 » The Genera of Fungi. iv, 496 .
New York, The H. W. Wilson Co.
A compendium designed to give a key to all genera of fungi known
up to 1930 and whose species are listed in Saccardo's Sylloge Fungorum.
This manual is unwieldy, has some errors based on increasing knowledge of
some of the greater groups, and has many incomplete or uncritical key
characters.
Coker, W. C., 1923. The Saprolegniaceae 201. Chapel Hill, The University
of North Carolina Press .
A well illustrated monographic study of the commonest or easiest
isolated family of water molds, the Saprolegniaceae.
Cooker, W. C. and V. D. Matthews s 1937. Saprolegniales . North American
Flora 2: 15-67 .
The definitive monograph of the Saprolegniales as known at present.
In this work certain concepts presented in the preceding reference are
clarified, but it is not illustrated and the two should be used together.
Cooke, W. B.s 1954a. Fungi in polluted water and sewage. I. Literature
Review. Sewage and Industrial Wastes 26: 539-549-
A review of the papers available at the time on fungi in polluted
water and sewage.
Cooke, W. B.s 1954b. Fungi in polluted water and sewage. II. Isolation
technique. Sewage and Industrial Wastes 26: 661-674.
A description of isolation techniques and a discussion of the
various media used in the isolation of fungi from polluted water and sewage.
Cooke, W. B., 1954c. Fungi in polluted water and sewage. III. Fungi in a
small polluted stream. Sewage and Industrial Wastes 26:
A quantitative study of the fungi found at eight stations on Lytle
Creek.
Garrett, S. D., 1956. Biology of Root-Infecting Fungi, xi, 293. New York,
Cambridge University Press.
A number of older concepts are clarified and some new concepts
given concerning the biology of root-infecting fungi and their occurrence
in the soil.
Gilman, J. C., 1945. A Manual of Soil Fungi, xi, 392, Ames, Iowa State
State College Press.
A compilation listing and describing all fungi known to occur in the
soil. Since the book is based on available descriptions and not on laboratory
experience of the writer, it indicates areas where additional work is required.
For instance, in certain larger genera one species may be described as it
appears on one medium, another from a different medium.
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Harvey, J„ V.» 1952. Relationship of aquatic fungi to water pollution.
Sewage and Industrial Wastes 24: 1159-1164.
A study based on the isolation of water molds from polluted streams
in the Little Miami Basin over the period of a year. It appears that few, if
any, aquatic fungi are definitely associated with polluted water.
Hesseltine, C. W.p 1953. Study of trickling filter fungi. Bull, Torr. Bot. CI.
80: 507-514.
Geotrichum candidum9 Fusarium aquaeductuum and Subbaromyces
splendens are consiclered to be the fungi of most importance on trickling
KTFersTtreating pharmaceutical and chemical wastes at Lederle Laboratories,
Pearl River, New York. Other fungi appearing in isolations were considered
contaminants or only incidentally present.
Hesseltine, C. W.s et al„, 1953. Coprogen, a new growth factor present in
dung, required by Pilobolus species. Mycologia 45: 7-19.
A description of the new growth factor coprogen, an indication that
it can be produced by other organisms in the habitat, and its effect on Pilobolus
Hughes, S. J., 1953. Conidiophor es, conidia and classification,,
Canadian Journ. Bot. 31: 577-659.
A description of a new system "of classification for the Fungi
Imperfecti based on methods of spore production.
Kolkwitz., R„, 1950. Oekologie der Saprobien. Uber die Beziehungen der
Wasserorganismen zur Umwelt. Schrift. ver. Wasser.-,
Boden-, und Lufthygiene . 4: 1-64.
An annotated list of species of organisms found in the several
zones of a stream identified in the Kolkwitz and Marsson system as:
polysaprobic , alpha-mesosaprobic , beta-masosaprobic and oligosaprobic .
Lodder, J. and N. J. W. Kreger-van Rij, 1952. The Yeasts. A Taxonomic
Study, xi, 713. New York, Interscience Publishers, Inc.
The definitive manual of yeasts. Classification into genera,
families and orders is based on morphology, into species and varieties is
^ased on reactions in culture to a number of standardized morphological
ind chemical tests .
^iddleton, J. T., 1943. The taxonomy, host range and geographic distribution
of the genus Pythium. Mem. Torr. Bot. CI. 20(1): 1-171.
The definitive manual for identification of the difficulFgenug Pythium
vhich occurs in soils and water. It is isolated occasionally on hemp seed.
>ut without knowledge of the species of plant with which the parent strain
vas associated it is difficult to make an identification.
^oser, M.s 1949. Untersuchungen Uber den Einfluss von Waldbranden auf
die Pilzvegetation I. Sydowia 3_: 336-383.
A study of burned areas in the Austrian Tyrol near Innsbruck in
fhich the concept of four degrees of relationship of a fungus or other
rganism to its environment is introduced.
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Raper, K. B. and C. Thorn, 1949. A Manual of the Penicillia. ci, 875.
Baltimore s The Williams and Wilkins Co.
The definitive manual for the genus Penicillium. This is most
useful in our work since of the 150 species described on standardized media,
about half have been isolated during the course of our work.
Seaver, F. J.s 1928. The North American Cup Fungi (Operculates). pp. 284.
New York, Published by the author.
A manual for the study of the Operculate Discomycetes , cup fungi.
Smith, G., 1954. An Introduction to Industrial Mycology. Fourth Edition.
xiv. 378. Londont Edward Arnold (Publishers) Ltd.
A manual for workers and students of industrial mycology. In this
book the commoner species used in industry and the commoner contaminants
of the laboratory and of industry are described. There are valuable chapters
on various phases of the biology and use of fungi.
Sparrow, F. K., 194S. Aquatic Phycomycetes , exclusive of the Saprolegniaceae
and Pythium. xix, 785. Ann Arbor, The University of
Michigan Press.
A manual for the study of various groups of aquatic fungi. This
will be of help to one who isolates fungi from waters especially if they
are associated with plankton, pollen, and other materials in the water. It
does not overlap the work of Coker or Middleton mentioned above.
Suter, R. and E. Moore, 1922. Stream pollution studies. State of New York
Conservation Commission, Albany, pp. 1-8, pi. 9.
This little pamphlet indicates that certain aquatic fungi can be
used as indicators of pollution but identifications, and therefore groups
mentioned appear to be in error.
Thorn, C. and K. B. Raper, 1945. A Manual of the Aspergilli. ix, 373.
Baltimore, The Williams and Wilkins Co
The definitive manual for the identification of Aspergillus . Species
in most of the subgeneric groups have been isolated from polluted water and
sewage and use of this manual makes their identification relatively easy.
Wilson, J. N., 1949. Microbiota of sewage treatment plants and polluted
streams, pp. 1 -15 , A .A .A ,S. symposium: Limnological
Aspects Water Supply and Sewage Disposal.
This paper presents certain basic concepts on the composition of the
slimes on trickling filters. While admitting the presence of fungi, no attempt
is made to determine the comparative role of any one or another species and
of bacteria in the formation of the basic slime.
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USE AND VALUE OF BIOLOGICAL INDICATORS OF POLLUTION:
FRESH WATER CLAMS AND SNAILS
William Marcus Ingram
Robert A. Taft Sanitary Engineering Center
U. S. Public Health Service
Cincinnati, Ohio
I. INTRODUCTION
In discussing fresh water clams and snails (mollusks), not enough
is known yet about molluscan ecology to name any species a pollution
indicator. There are mollusks tolerant to certain effects of pollutants
such as septicity, but even these are not pollution indicators. Species
that are found associated with domestic sewage in septic reaches of
water, as Musculium transversum, Pisidium idahoensis, Physa integra,
and Physa heterostropha, are also found in high dissolved oxygen areas
of lakes and streams unpolluted by domestic sewage or putrescible
industrial wastes.
On the other hand certain mollusks, such as the Unionidae**-, are
not associated with near-septic-water resulting from pollution These
have an index value in that their presence typically indicates good
dissolved oxygen and attendant physical and chemical conditions associated
with unpolluted water. Such mollusks can be called clean water index
organisms.
Apart from systematic morphological studies, it is not realistic
to isolate a single group of organisms such as mollusks from other
animals and plants that are associates under similar ecological conditions
in clean or polluted water. It is the study of the total biota which tells
one most about water conditions.
Members of the family Unionidae have had various common names
applied to them: Mussels, fresh water clams, and naiads.
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In this respect, the presence of an assemblage of rat-tailed maggots,
Eristalis tenax; sewage mosquitoes, Culex pipiens; sludge worms,
Tub if ex tubifex; blood worms, CThironomus plumosus; physid snails,
Physa Integra; and finger-nail clams, Musculium transversum, and an
absence of Unionidae, mayflies, caddis worms, stoneflies, and shiners
would indicate to investigators stream reaches highly degraded by
domestic sewage, for example. Thus, certain associations of organisms
that tolerate such pollutional conditions as septicity and the absence of
intolerant forms can be looked upon collectively to form pollution
tolerant biological assemblages, even though any single mollusk species
or other members of the assemblage may not be called a pollution
indicator. The presence of intolerant mollusks, with other intolerant
animals, lend themselves usefully in sanitary science to establishing
parameters around areas of septicity and sludge deposits resulting from
domestic dewage.,
Information is not available that can be presented to indicate that
various species of mollusks can be used to indicate varying degrees of
water quality, i. e. from high dissolved oxygen values by gradations to
septicity, such as can be measured by chemical tests,, Also, various
species cannot be used to measure variations in fecal contamination as
can certain bacteria,
The majority of studies made in United States waterways dealing
with the effects of pollution on mollusks are related to domestic sewage.
The principal effect of such pollution on water quality, investigated in
relation to mollusk survival, is that of lowered dissolved oxygen„ Some
attention has also been given to the effects on mollusks of bottom deposits
attendant to domestic sewage and silt pollution. Little information
dealing specifically with the effects of industrial wastes or their components
on fresh water mollusks has been fourad.
The information presented below can assist those working with
biological indices of pollution to group mollusks as either pollution-
tolerant or clean-water forms. Consideration is given to the following
aspects of this subject: references relating mollusks to pollution;
structural and life cycle variations relating to survival in polluted
water; natural variations in distribution not related to pollution; and
identification sources.
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II. DISCUSSION OF SELECTED REFERENCES
Selected references that may be readily available to those work-
ing in sanitary sciences are cited here that especially deal with water-
ways of the United States. No attempt is made to present a complete
literature review covering pollution and its effects on mollusks. Many
of the included references should point out to those studying bottom
organisms the importance of recording chemical and physical data
that can be analyzed in relation to tolerances of specific mollusks to
pollutants.
Available literature relating mollusks to water chemistry is woe-
fully lacking. When the word "pollution11 is used, the general inference
is to domestic sewage. Except in a few specific studies of industrial
wastes, cognizance often is not taken of the effects of such wastes in
association with domestic sewage, even though they may have been
related to the presence or absence of mollusks.
In order to make data concerning the effects of pollution on
mollusks comparable, pollution should be defined both chemically and
physically. It is also necessary to identify mollusks to species, if
pollution-tolerant ones are to be exactingly separated from intolerant
ones. Specific identification is particularly important to those who
hope to find indicators of degrees of pollution.
If work, under field conditions, on the relationship of mollusks
to physical and chemical factors is contemplated, Boycott (1936)
should be consulted early in the planning stages. Even though relatively
few of the species he deals with are found in North America, the
information he presents associating mollusks with water chemistry should
provide valuable background information for North American studies.
(1) References Relating Molluska to Pollution in General
The following references relate tnollusks to pollution in general
without consideration of chemical and physical data. Such papers are
valuable, in that they contain references to mollusks already identified
to genera or to species by outstanding authorities in Conchology, By
being aware of such references aquatic biologists working oil .water
pollution problems have mollusk names available from certain areas,
that may give them a lead to identification of current collections.
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In relation to water pollution in general with special reference
to streams in western Pennsylvania, Ortman (1909) wrote that the
Unionidae are the first to be eliminated from polluted waters, Further,
he states that the genera Pleurocera, Goniobasis, and Anculosa are
usually absent in polluted rivers, but were found surviving when the
Unionidae and fishes were, for the greater part, gone from the
Allegheny River in Venango County, Pennsylvania. The genera
Lymnaea, Physa, and Planorbis are noted to be more resistant because
they are air-breathers. Physa is the hardiest and is stated to be a
genus ". . . which represents in certain instances the only remaining
life in certain rivers. But there also seems to be a limit to its power
of endurance, and in very badly polluted streams also Physa is absent. "
Baker (1911), in quoting French investigators, states that
Sphaerium, Pisidium, and Planorbis resist the effects of water contam-
inated with sewage, oil, and chemicals better than Lymnaea. Baker (1911)
reports, from his own observations made at Rochester, New York, that
the Genessee River into which sewage has been discharged for the past". . .
ten or fifteen. . . years is . . . at the present time ... of the consistency
of dirty, greasy dish water, yet Galba catascopium and Planorbis
trivolvis live and thrive by thousands in this seemingly unfavorable
environment. The writer's observations have been that chemicals and oil
are deadly to molluscan life, while sewage does not materially affect
them. " In a footnote to this statement, Baker comments that since writ-
ing the above, sewage in the Genessee River has become . .of such a
highly concentrated form that the mollusks have all disappeared in the
river for a mile or two below the point of discharge into the river. "
In a second report on the pollution of the Genessee River at
Rochester, New York, Baker (1922) states that he has studied its
pollution for 27 years, from 1892 to 1919. He mentions that pollutants
are "... sewage . . . discharged into the river in a crude condition . . , "
and that "refuse and other waste matter, both liquid and solid, also
enter the stream from gas works, tanneries, and manufacturing plants.
Mollusk collections that he made in 1892, before pollution became appar-
ent, represented 9 species: Musculium transversum, M. partumeium,
Bythinia tentaculata, Planorbis trivolvis, Physa gyrinaT"P. sayii,
P. oneida, Galba caperata, and G. catascopium. In 1907~the above
species of Musculium and Bythinia had disappeared^ with the air-
breathers Planorbis, Physa, and Galba still present but reduced in numbers,
In 1910 all mollusks had disappeared and none were found in subsequent
collecting trips from 1910 to 1913. Baker (1922) describes studies that
G. C. Whipple made on the river in 1912 after molluscan life had disap-
peared. He reports that Whipple found the dissolved oxygen varying
from 5 to 41 per cent of saturation in August.
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On one day in this month saturation did not exceed 5 per cent in
a 3 mile reach from the surface to bottom in a depth of about 26 feet.
He states that in 1917 a large part of the Rochester sewage, 32 million
gallons a day, was diverted from the river to a sewage treatment
plant, the effluent of which was discharged into deep water of Lake
Ontario. In 1919 Baker reports the following mollusks occupying the
reach of stream that had become devoid of them before sewage treatment
was installed: Musculium transversum, Bythinia tentaculata, Galba
catascopium, Planorbis trivolvis, Physa integra, land P. oneidal
According to the study by Wilson and Clark (1912) on the mussel
fauna of the Kankakee Basin in relation to destruction by dredging
operations, "The most fatal condition is the constant movement of the
fine sand and silt al6ng the bottom of dredged channels. " They further
state, "Portions of the basin which were dredged 15 or 20 years ago
show no signs of restocking with mussels, though there are thousands of
them close at hand in old channels. "
Considering the effects of pollution on the mussel fauna of the Big
Vermilion River and its tributaries in Illinois Baker (1922) states
that, "sewage pollution has killed all clean water life for a distance of
fourteen miles below Urbana and has made the stream an unfavorable
environment for a distance of twenty miles,, Below this point the fauna
is normal and is not affected by sewage pollution,, " He observed that of
that large species of Unionidae, Amblema undulata and Lasmigona
complanata.resisted pollution conditions better than others„ In a pre-
liminary paper to this report, Baker and Smith (1919) also wrote on
the same subject,,
Baker (1928), in the pelecypod part of his monograph on Wisconsin
fresh water snails and clams, mentions that stream pollution by sewage
and manufacturing wastes produces unfavorable conditions for mollusks.
In reference to industrial wastes, he writes that coal tars and oils in
particular quickly make a stream totally unfit for any kind of animal life.
In his work on the mollusca of Michigan Goodrich (1932) states
that Lymnaea stagnalis appressa are being reduced by drainage enter -
prises and pollution. He adds that this gastropod probably disappeared
from great areas in a few years because waterways were used for logging
purposes and sawdust disposal.
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Van der Schalie (1936) notes that over the years domestic sewage
and industrial wastes in general, along with other factors, have had a
detrimental effect on the naiad fauna of the St. Joseph River Drainage
in southwestern Michigan. Effects of pollution on specific mollusks are
not discussed.
Van der Schalie (1936a), when discussing factors contributing to the
depletion of naiades in the eastern United States in relation to the fauna
of the Mississippi River, states, "Pollution, particularly below the
several large cities located on the river and its important tributaries,
was responsible for a heavy mortality in glochidia which were attached
by bacteria and infusoria. " In discussing Ellis' (1931) field investigations
relating to the effects of silt on the fisheries of the Mississippi River,
he further states that "The river is practically devoid of mussels from
the region of St. Louis, Missouri, to its mouth, a condition accounted
for by the tons of silt carried to the stream and deposited in it by the
waters of the Missouri River which enters the Mississippi near St.
Louis. ... . " Writing about the "South Atlantic Costal drainages" he
mentions that mine wastes from coal mines in the headwaters of the
James River have damaged the fauna of this stream. With reference
to the mussel fauna of the Great Lakes Drainage rivers in Michigan
he comments that industrial wastes and sewage are particularly damaging
to mussels in the Saginaw drainage, St. Joseph, the Kalamazoo, the
Grand, and Rouge Rivers; wastes from beet sugar refineries are felt
to be responsible for "unproductive" areas in the Raisin and Pine Rivers.
In reference to the above drainage area he states, "In many localities
action has been taken by the State to curtail such destructive influences
(pollution), though usually much irreparable damage has been done
before preventive action has become effective. " Thus, in relation to
the effects of factors affecting mussels of the eastern United States
van der Schalie writes, ". . . silting, pollution by sewage, mine and
industrial wastes, power-dam developments and unrestricted mussel
gathering for the pearl button industry, have resulted in the critical
depletion of the formerly abundant Naiad fauna. "
In writing about the depletion of the mussel resources of Michigan
van der Schalie (1938a) mentions that among the outstanding factors are
pollution in streams by sewage and industrial wastes and the extensive
program of power dam developments. He further comments that mussels
are among the most sensitive of organisms to pollution and are among
the first to perish where pollution is in evidence.
Van der Schalie (1938b) points out that the Cahaba River in
northern Alabama, as of 1938, was unusually productive conchologically,
but that there are several potential dangers to Cahaba mussels:
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the possibility of acid pollution fronri the Cahaba coal field, industrial
waste concentrations and sewage from Birmingham, and dam
construction. He writes, "In view of these possible changes, studies
of the fauna under natural conditions are highly desirable. "
Goodrich (1939) mentions that the lumber industry has affected
mollusks by fire, sawdust and rafting. However, he does not assess
damages.
From mussel studies of the Grand and Muskegon Rivers in
Michigan van der Schalie (1941) writes that many factors are causing
the depletion of the indigenous fauna. Thus, it becomes increasingly
important to gather ecological and distributional data about them
before damage becomes too severe. Factors in addition to dredging,
that have damaged mussel populations in the Grand River, are many
kinds of industrial wastes and sewage in reaches below Jackson,
Lansing, and Grand Rapida. Gathering mussels with apparatus injur-
ious to mussel beds is also mentioned. In relation to the Muskegon
River, he lists power dams and pollution as being detrimental to
mussels and hindering their distribution through obstructing fish,
the " carrying agents for fresh-water mussels. "
Van Horn (1949) associates the air-breathing snails Physa and
Planorbis with a zone of recent pollution, and states, "In this zone also,
as the oxygen concentration is decreased, one may find sewage fungi
.. . such as Sphaerotilus, Leptomitus, Thiothrix, and others. "
According to van der Schalie and van der Schalie (1950), report-
ing on the mussels of the Mississippi River, "The several surveys of
the Mississippi emphasize that the changes brought about in this
drainage through adverse conditions, such as silting, pollution,
intensive exploitation of the mussel fauna, power dam construction,
itc. , are all tending to alter decidedly, as well as to reduce, the
Jriginal fauna. "
Beck (1954), in his ecological classification of the streams of
Florida relating organisms to water pollution surveys, refers mollusks
>f the genera Physa and Ferrissia and the Sphaeriidae to a "Class III"
vhich includes organisms which have been found in heavily polluted
ireas. He comments that no organisms in this class may be considered
ndicative of pollution, since organisms contained therein may be found
n clean, moderately polluted, or grossly polluted water. Goniobasis
pp. is placed in a "Class I" containing organisms that have been found
o tolerate no appreciable organic pollution, "... the more sensitive
orms. " He states that, The presence of class I organisms is
onsidered to indicate that the water in which they are found is clean. "
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Car lander (1954) writes about the general effects of pollution,
including silt, on the mussels of the upper Mississippi River using
material published by the U. S. Fish and Wildlife Service as a basis
for her discussion.
In presenting his list of mollusks (Table-1) that can survive
"....at least to some degre..." in zones of degradation and recovery,
Wurtz (1956) points out that " we are woefully lacking in knowledge
on this subject." He further writes that the exact tolerance limits of
the mollusks he lists are not known, and that as far as he can ascertai
no mollusks are able to withstand protracted gross pollution. From
among the 34 species and subspecies of mollusks he has considered,
he states that Physa heterostropha is the most tolerant species that
has been found.
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TABLE I*
Mollusks Reported to Survive "at least some degree"
in Zones of
Degradation and Recovery
GILL-BREATHING SNAILS
Family - Fhysidae
FRESH WATER CLAMS
(Ctenobranchiata)
1. Physa gyrina
Family - Sphaeriidae
Family - Viviparidae
2. P. heterostropha
1. Sphaerium rhomboideum
1. Campeloma integrum
3. P. integra
2. S. corneum
2. C. rufum
4. Aplexa hypnorum
3. S. striatinum
Family Amnicolidae
Family - Planorbidae
4. S. sulcatum
1. Bulimus tentaculatus
1. Helisoma anceps
5. S. (Musculium) securis
2. H. trivolvis
6. S. (Musculium) tranversum
LUNG-BREATHING
3. Gyralus arcticus
7. Pisidium amnicum
(Pulmonata)
4. Menetus dilatatus
8. P. casertanum
Family - Lymnaeidae
Family - Ancylidae
9. P. compressum
1. Lymnaea caperata
1. Ferrissia fusca
10. P. fallax
2. L. humilis
2. F. tarda
11. P. henslorvanum
3. L. obrussa
12. P. subtruncatum
4. L. polustris
Family - Dresisseniidae
5. L. stagnalis
1. Mytilopsis leucophaeatus"
6. L. auricularia
Family - Mactridaex
7. Pseudosuccinae columella
1. Rangia cuneata
* Mollusks of Wurtz (1956) that can survive . . at least to some degree ..." in zones of degradation
and recovery. Wurtz's data have been organized to form this table by W. M. Ingram.
Only one species in each family in the United States; the former with a range from Maryland to
Florida, and the latter with a range from Alabama to Texas.
-------
(2) References Relating Mollusks to Specific Aspects of Water Quality
Associated with Pollution.
The following investigators have presented varying amounts of
physical and chemical data that relate mollusks to certain aspects of
water quality.
Juday (1908 and 1921) reports a Sphaeriid clam, Corneocyclas
[ =Pisidum] idahoensis, from septic water in Lake Mendota, Wisconsin;
however he does not mention specific pollutional conditions. From
laboratory observations he concluded that this clam may remain quiescenl
with its valves closed for as long as three months each summer in the
septic bottom ooze of the lake. He found that clams surviving in septic
water with their valves tightly closed became active when placed in
aerated water. Shelford (1913) has listed Juday's (1908) work relating
to survival of Pisidium idahoensis under anaerobic conditions.
Weston and Turner (1917) found Physa heterostropha, Helisoma
trivolvis, Segmentina armigera, and Unio"complanata living in a stream
below a sewage treatment plant outfall where average monthly dissolved
oxygen values in October, June, July, and August were between 1 and
2 p. p. m. These writers do not present D. O. figures other than in
monthly averages. The gastropod Campeloma decisum was not taken in
any stream reaches where the average monthly D. O. was below 5. p. p.m.
Jewell (1920) discusses mollusks and other organisms associated
with clean water and polluted water reaches of the Sangamon River,
Illinois. This author mentions that there is some domestic sewage
pollution from the effluent of a septic tank in the town of Monticello
entering the clean water area, but such pollution only affects the water
in a restricted area. Chemical conditions of clean water reaches are
stated to be, ". . . alkalinity to methyl orange of 222 p. p. m. and a pH
of 8. . . transparent to a depth of a foot or more. 11 Bottom materials
are said to consist of gravel. Close to shore it was observed that a
". . . layer of organic deposit covered the bottom. . . . " Mollusks
associated with pools in such water were the Unionidae species,
Lampsilis luteola, Quadrula pustulosa, Q. undulata, Q. rubiginosa,
^fritigonia tuberculata, Symphynota costata, Strophitus edentulus, and
Anodonta~grandis, unnamed species of Sphaeriidae, unnamed species of
fleuroceridae, and Campeloma subsolidum.
Below the reach of the Sangamon described above, sewage of Decatur
is discharged. Bottom animals were stated to disappear immediately
and . .not until Noantic is reached, about 30 miles below
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[Decatur], do the first animals appear, " No mollusks are listed in
Jewell's tables for 75 miles below Decatur,, In this distance figures
are listed showing variations in chemical and physical conditions:
D. O. from 0 to 4, 1 p. p. m. alkalinity from 260 to 348 p. p. m„ , pH
7 to 8, odor from septic and putrid to not pronounced, appearance
from sewage to inky to milky to grey to turbid to green-grey. At
the 75-mile station where mollusks are first noted to reappear, chemical
and physical conditions show; D. O. of 7. 8 p.p. m. , an alkalinity of Z87
p. p. m. , a green appearance, and no odor. The mollusks occuring at
this station were Unionidae, Quadrula undulata, Q. lachramosa, Lampsilis
alata, L. luteola, L. anadontoides, Unio gibbosus, Anodonta grandis,
'Tritigo'nia tuberculata, Strophitus edentulus, .MasmoSonta costata,
unidentified "Sphaerium, 11 and the gastropods Pleurocera elevatum and
Campeloma subsolidum. Unionidae and gill-breathing snails at various
stations are associated with disolved oxygen conditions that were not
less than 7. 5 p, p. m.
Richardson published a number of papers relating general pollution
of the Illinois River to bottom fauna, among which his 1921, 1925, 1925a,
and 1928 papers are cited in the bibliography as being of special interest
to those studying mollusks. Richardson's 1925 paper lends itself to an
attempt by the writer to generally correlate dissolved oxygen-mollusk
relationships at certain stations in the reach from Chillicothe into Spring
Bay Narrows, from the few dissolved oxygen figures that he presents
under the title, "Bottom Dissolved Oxygen Mid Channel. n Three species
of Sphaeriidae, Musculium transversum, M. truncatum, and Pisidium
complanatum, can be associated with a minimum D. O. of 0, 2 p„ p, m.
The gill-breathing snail, Valvata tricarinata can be associated with a
minimum D. O. of 1. 4 p. p. m. The gill-breathing snail, Campeloma
subsolidum, which Richardson stated as being a survivor under poor
dissolved oxygen conditions, can be associated with dissolved oxygen
figures between 0. 5 to 3. 0 p. p. m.
Baker (1926) presents a review of Richardson's 1925 papers on the
effects of pollution on bottom organisms in the Illinois River. Data from
the review state, "It is shown prior to 1915 there have disappeared from
Peoria Lake (a widening of the Illinois River) about forty species of river
mussels (only three or four species are left), all of the small snails be-
longing to the water-breathing [sic. = gill-breathing] family Amnicolidae,
all but one species of the large water snails belonging to the Viviparidae,
the remaining species being recorded as Campeloma subsolidum, and a
varied weed fauna. There remains certain species which appear to be
more tolerant of pollutional conditions. These are Musculium transversum,
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Pis-iwiac. jo-:iprcGsuin, P. paupcrculum crystalense, Campeloma
suisoii^uin, aiid Cpnaerium s tannine urn. Several species less tolerant
to sewage conditions were observed "in peculiarly favorable conditions,
usually in strong current in midchannel or elsewhere where oxygen
conditions were good. These included Anodonta imbecillis, an insect
Corixa, a caddis-fly larva (Leptocerid^ Goniobasis livescens,
Pleurocera elevatum lewisii, Quadrula plicata ( - peruviana), Hyalella,
a sponge, and a Hydropsyche. These species, however, were observed
to vary in presence during different years. "
Suter and Moore(l922) state that Physa heterostropha may be found
in septic regions and list it with Pi;sidium abditum, Goniobasis
virginica, and Campeloma decisum as an organism tolerant of pollution;
they do not mention that the latter three species can survive septicity.
Turner (1927) lists Physa heterostropha as found in septic water
and Planorbis panus from stagnant water and the vicinity of sewer out-
lets. Campeloma decisum is described as being more sensitive than
the other species since ". . . . it seems to thrive under clean water condi-
tions. "
Wiebe (1928), in his paper on the effects of pollution on the upper
Mississippi River, collected Helisoma trivolvis, Musculium transversum
and Campeloma integrum on August 27 at his Red Wing station with the
D. O. at 2. 83 p. p. m. ; of 22 D. O. measurements made in August the
average at this station was 2. 25 p. p. m. with a minimum of 1.12 p. p. m.
and a high of 4. 01 p„p.m. Individuals of Musculium (near) transversum
were collected at his Jackson Street station on August 17 with the D. O.
at 0. 87 p. p. m. ; of 22 D. O. measurements made in August the average
at this station was 0. 87 p. p„ m. with a range from 0 to 2. 52 p. p. m.
His data on tables 4 and 5 indicate that mussels (not identified) were
taken in September at the Jackson Street station when the D. O. was
5. 73 p. p. m. with the range of 20 readings for September varying from
0C 44 to 8. 14 p. p. m. He records collections of Anodonta imbecillis
on September 17 at the Red Wing Station with the D. O. on this date
being 4. 39 and a range of 21 readings for September varying from 2. 89
to 6.44 p. p. m. Campeloma rufum was taken at two stations where the
D. O. was always above 5 p. p. m. for 61 measurements; Wiebe never-
theless comments in relation to this species, . very likely it is
one of the more tolerant forms;" this conclusion is based on the fact
that 1, 600 specimens per square yard of bottom were taken some 5Q
yards below a sewage outfall at one station and were associated with
15, 120 Tubificidae and 54 Sphaerium notatum per square yard.
Pleurocera acuta is reported from two stations where the D. O. was
always above 4. 30 p. p. m. for 65 measurements.
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Purdy (1930), in summarizing pollution data on organisms other
than plankton in his Illinois River work, states that the Sphaeriidae
are often very numerous in moderately polluted water. He further
writes that these mollusks cannot stand "extreme" conditions that
Tubificid worms can and that they will die when oxygen becomes
largely depleted. He states that, "Apparently their Sphaeriidae
large numbers in places where water is polluted is a question of
their abundant food supply of microscopic organisms normally found
there„ " Purdy's data for the Illinois River, as correlated by Ingram
with those of Hoskins et. al. (1927), show that unidentified Sphaeriidae
were collected at Chillicothe where the dissolved oxygen was recorded
as low as 1.23 p. p. m. in August. The highest D. O. at this station
was 7„ 79 p. p. m. in February. Unidentified air-breathing snails
collected by Purdy (1930) at Lockport, as correlated with Hoskins1 s
et. al. (1927) data, were taken at this station where septicity existed
in August and the D. 0. was 9. 11 in February. The pollution in the
Illinois River at the time the above data were collected was from
numerous industrial wastes and domestic sewage.
Ellis (1931a) writes that juvenile and young mussels are quite
sensitive to oxygen reduction and that adult mussels . . usually become
inactive when the oxygen tension of the water is reduced to 20 per cent
saturation or less. " He emphasizes the detrimental effect of erosion
silt on clams. In relation to the general effect of industrial wastes on
¦nussels, he writes, "Whenever concentrated industrial wastes are
>oured into the streams the fresh-water mussels suffer because of their
nability to change location quickly and because of the ease with which
he blood of fresh-water mussels takes up the various substances in
he surrounding waters. ..."
Van der Schalie (1938) has presented chemical and physical data
ssociated with unpolluted water and water polluted by domestic sewage
n. relation to the distribution of Unionidae in the Huron River, Michigan.
Leferring to the effect of pollution on mussels, he states, "Below Ann
irbor, sewage has been very detrimental to the fauna. It is true
tiat sewage may, if not too concentrated, increase the productivity of
ections of a stream by increasing the dissolved organic compounds,
ut below Ann Arbor and as far as the backwaters of Geddes dam,
ollution is so concentrated that it has killed all Naiades. Furthermore,
aere is such a heavy deposit of sludge in this zone that it will be many
ears before bottom conditions will permit the re-establishment of a
luna even though the discharge of sewage into the river be discontinued. "
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Goodrich (1945), in his monograph on the gilled-snail Goniobasis
livescens in Michigan, associates water chemistry with its occurrence
and with Pleuroceridae in general. He writes that this species has
disappeared from the heavily polluted Huron River. He associates
massive kills of Pleuroceridae in western Lake Erie with exhaustion
of oxygen under ice cover. G. livescens has been found in streams
with the alkalinity ranging from 87 to 233 p. p. m. He states that
a drop in pH much below 7. 0 does not permit survival of pleurocerid
life judging by studies made by C.S. Shoup in the Obey River in 1939.
Streams with a pH above 7. 0 were extensively colonized, while these
mollusks were absent in water with a pH of 6. 1. Members of the
genus Goniobasis in general endure periodic silting of streams that
accompany freshets. Certain ones, G. virginica for example, may
live in tidal reaches of streams where the salinity is 50 per cent of
that of sea water.
Whipple, Fair, and Whipple (1949) present a list of mollusk
species that they have associated with the pollution zones of Kolkwitz
and Marsson (Table 2). They list no mollusks in the polysaprobic
zone that they partially characterize by the presence of black sludge
accumulations on the bottom and a lack of oxygen. They note that
life in the mesosaprobic zone is commonly tolerant of dilute or
imperfectly purified sewage and its products of decomposition, and
state that "Many bacteria are still present. " Pertaining to the
oligosaprobic zone they write, "This is a zone of cleaner water
in which mineralization has been completed. ..... The water is
practically saturated with oxygen, sometimes even supersaturated. "
It should be noted from Table 2 that many mollusks associated
with the mesosaprobic zone are also associated with the oligosaprobic.
One Unionid clam and certain gill-breathing snails of the genera
Campeloma, Viviparous, and Valvata that commonly inhabit clean
water are associated with the mesosaprobic zone.
Patrick (1949) associated Physa heterostropha with "polluted
water" conditions at a station 105 in Lititz Run, Pennsylvania.
Chemical data are presented in tables, separated from biological
data; such information presented for August, a month indicated to
be "low water" for this station, shows a pH of 7. 3; D. O. , 8. 00 p. p. m.
turbidity, 10.8 p. p. m. ; Nitrogen (N as NH_), 0.015; Nitrogen
(N as N02), 0. 0260; Nitrogen (N as NOj), i" 3. 120; etc.
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TABLE 2
Mollusks Related by Whipple, Fair, and Whipple to
Pollution Zones of Kolkwitz and Mars son
Pollutjonal Zones of Kolkwitz and Mars son
~~ Kdollusks
Mesosaprobic
family - sphaeriidae
Pisidium amnicum
" compressum
" fossarinum
" pauper culum
Sphaerium corneum
11 moenanum
11 vivicolum
" stamineum
" striatinum
Musculium transversum
11 truncatum
family - unionidae
Unio batavus
11 pictorum
" tumidus
Anodonta mutabilis
family - margaritanidae
Mararitana margaritifera
TOtlLY - PMY51DAE
Aplexa hypnorum
Physa acuta
" fontinalis
Family - lymnaeidae
Lymnaea auricularia
" ovata
" palustris
" peregra
" stagnalis
family - helisomidae
Planorbis carinatus
" corneus
" marginatus
ANCYLIDAE
Ancylus lacustris
fluviatilis
family
Family
- VALVAT1DAE
Valvata piscinalis
11 tricarinata
Family - viviparidae
Viviparus contectus
" fasciatus
Campeloma subsolidum
Oligo
sap-
robic
Poly
sap-
robic
x
X
X
X
X
x (indi
x(indi:
x
;ferent
ferent'
X
x
x
X
X
X
X
X
X
X
X
No sub-
design-
nation
x
X
X
X
Alpha
Beta
x
x
x
X
X
X
X
X
X
X
X
X
r
&
w
4
fD
fu
?
&
$
tu
X
X
X
X
X
* Not allspecies are presented here (arranged by W. M. Ingram)
>#9.
OQ p
£ H
¦ &
-108-
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Gaufin and Tarzwell (1952) reported Physa Integra from all
stations on Lytle Creek, Ohio, during studies conducted in May
and August, and list it as abundant at a station where D. O. was
recorded as low as 0.2 p. p. m. Sphaerium solidulum, as reported,
was not collected at stations where the D, O. was less than 4. 5
p. p.m. as based on diurnal sampling.
On the basis of collections from Lytle Creek Ingram, Ballinger,
and Gaufin (1953) report Sphaerium solidulum intolerant of pollution
from domestic sewage including septicity and sludge deposits, and
intolerant of bottom areas covered with zoogleal organisms. Certain
literature relating to the tolerance of the sphaeriidae to pollution is
discussed.
Gaufin and Tarzwell (1955) show that during January and February
of 1952 Ferrissia rivularis, Sphaerium solidulum, and Pisidium
casertanum were not taken in Lytle Creek reaches that had had a
septic record in August of 1951, even though the minimum winter
D. O. was above 7.0 p. p. m. They present data for October of
1951 showing that Ferrissia rivularis, Musculium transversum,
Pisidium casertanum, and Lymnaea humilis modicella also were
not collected from reaches that had a septic record in August of
1951. A few Sphaerium solidulum were collected from a station
that had had an August septic record.
(3) References Relating Mollusks to "Wastes from Specific,
Industrial Operations
Few available references discuss mollusks in relation to
specific wastes from industrial operations in natural waters.
In Culter's (1930) work concerning the blanketing effect of pulp
and paper mill wastes in Ticonderoga Creek, New York, he states
that Campeloma decisum was abundant where pulp was the thickest.
In addition, the following mollusks are listed in a table as being
associated with "a pulpy bottom 8 inches thick . . . Amnicola limosa,
Planorbis antrosus, Calyculina hirsutus, Lymnaea decidiosa an3
Sphaerium fabale. Sphaerium striatinum is listed as occuring on a
stream bottom covered by pulp up to one inch thick. Cutler states
that at a station in Lake Erie at the mouth of the creek there was no
molluscan life; by inference, he attributes this to the drifting action
of pulp. At a second station in Lake Erie, also at the mouth of the
creek but protected from pulp deposits, he reports the occurrence
of the following mollusks: Valvata tricarinata, Amnicola limosa,
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Bythinia tentaculata, Calyculina securis, Planorbis campanulatus,
Planorbis hirsutus, Lymnaea (Acella) haldemanni, Lymnaea
decidiosa, Sphaerium stiratinum, and Sphaerium fabaleo No
consideration was given to possible effects of toxicity on m
-------
Table 3
pH RANGES ASSOCIATED WITH SELECTED MOLLUSK GENERA (AFTER MORRISON 1932)
pH RANGE
GENERA
sm///m/m/mmi///m rmm////////m////mA
WtMftftfWMWlWih 7T7////l/////////////////h
V////////////////////ti///////////77777m
r///////. >mm)im/t/mi/mm\
K/////////////////////// Y77777////////////////m
wmnmnnnmiL
wm/mwmmmmt/t w//mm//////mm/771
mmnmmmimm
wzzzm
v/mmmm/m/i m//mtmm/mmm\
mmmmmifTTTTTrm
v/mmm//mm vmmmmwmmm
Vatvafa
Campe/oma
Am ni co la
Lymnaea
Stagn/co/a
Ace! I a
Bulimnaea
Fossaria
He/isoma
Gyrautus
Ferr/ssia
Physa
GILL BREATHING
SNAILS
LUNG BREATHING
SNAILS
NEUTRAL
VffMiiiiiiiiiil
Minium
////////////////A
Vfffff/fffffffffffz wm/NN/mmmumi
Mifiiiiiiiiiii/j/if/iiiM
//////////////////a
Wiiiiiiiiiiiiiifiitiifiito
vitffmmmtim/f/fj / ff/f/m/fffff/fmf/fffA
Muuuiuiimirrmrm,
if/f/ffmmnfftTrTT.ftfmmiuuiumimih
Fusconoria
Amb/ema
E Hiptio
Lasmigona
Anodonta
Anodontoides
Strophitus
Lamps/lis
Sphaerium
Muscu/ium
Pisidium
UNIONIDAE
CLAMS
SPHAERUDAE
CLAMS
-------
Such a range of pH values tolerated by mollusks is within that which
is listed by Doudoroff and Katz (1950) tolerable to most fresh water
fish without lethal effects. These writers state, "It appears that,
under otherwise favorable conditions, pH values above 5. 0 and rang-
ing upward to pH 9. 0 at least are not lethal for most fully developed
fresh-water fishes. " In addition, these investigators write, concern-
ing fish survival in relation to pH, "Much more extreme pH values,
perhaps below 4, 0 and well above 10. 0, also can be tolerated in-
definitely by resistant species. However, regardless of the nature
of acid or alkaline wastes responsible, such extreme conditions,
associated with industrial pollution, are evidently undesirable and
hazardous for fish life in waters which are naturally so acid. " Most
species of Unionidae, as reported by Morrison (1932) and dependent
upon fish for the completion of their life-cycles, were found in a pH
range upward from 6. 9. However, one member of the genus Anodonta,
A. marginata, was reported from water having a pH of 6. 3.
Jewell (1922) has reported mollusks living in the Big Muddy River,
Illinois, a stream characterized as a ". . .naturally acid stream. with
a pH range stated to be from 5. 8 to 7. 2, Jewell presents species
identifications of nine Unionidae from the stream. In relation to snails
found in this river she writes, "Abundant as were mussels in the Big
Muddy River, only two snails (individuals) were found: one a living
Pleurocera elevatum taken opposit Benton. . ., the other a Campeloma
subsolidum taken near Murphysboro. No dead shells were found to
indicate that snails had ever been present. " This writer states that
fish were". . . everywhere abundant. ", and that dogfish, sunfish and
lative carp were taken with hook and line, while large numbers of
minnows, cat fingerlings (at one time a school of several hundred), and
¦midentified fry were seen in pools and riffles. There were also
numerous swarms of top minnows, while gar, fifteen to eighteen inches
ttj length, were seen in ever-increasing numbers toward the mouth of
:he river. "
The writer has found Physa integra in sections of the Mahoning River,
Dhio, where the pH range over several days varied from 4. 1 to 7. 3„
Mollusks Associated with Sewage Treatment Installations
The information included here deals with species of Physa snails
n trickling filter beds of sewage treatment installations treating
lomestic sewage. A number of persons talking with the writer have,
stated that they have seen snails in trickling filter beds; however, very
ittle data are available in the literature to describe conditions under
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which snails live in such situations. Study of snails in trickling
filters offers ready access to those who wish to obtain data relating
snails to water not completely purified from the effects of pollution.
Certain chemical and physical tests, kept routinely at many secondary
sewage treatment plants employing trickling filters, can be used to
relate snails living in filters to specific ranges of water quality.
Brown (1937) studied Physa anatina living in sprinkling filters in
a sewage treatment plant at Urbana-Champaign, Illinois, during parts
of the years 1932-35. Brown writes that the plant has Imhoff tanks
and sprinkling filters, „ where jets of the sewage are forced into
the air for aeration. and a secondary settling tank. Part of the
final effluent is diverted into an experimental lagoon and part into
the Saline Drainage Ditch, a tributary of the Big Vermilion River.
Except for bacterial numbers no operational data are presented in the
paper. In reference to bacteria it states, "At the time crude sewage
enters the plant it contains 2, 100, 000 bacteria per cubic centimeter,
but when finally treated, the number has been reduced to 700 per cubic
centimeter,, " Brown (1937) believes that snails play a part in the
reduction of numbers of bacteria^ In addition to collecting Physa
anatina in the rock beds of sprinkling filters, individuals were taken
from the secondary settling tank and from the Saline Drainage Ditch
and experimental lagoon receiving the final effluent. During the course
of the study, in addition to Physa, 8 individuals of the snail Fossaria
modicella were reported from the secondary settling tank, but from
no other structures„
It is mentioned that in maintenance operations from 25 to 30
bushels of empty shells are removed each year in July and November
from a conduit of the secondary settling tank„ Brown believes that the
majority of snails in the sprinkling filter beds die each winter. Reported
observations based on shell size present evidence to indicate that life
cycles of snails are completed in the sprinkling filter beds„ Whether
snails occur through the depth of the beds is not indicated.
A 1929 Annual Report of the Urbana-Champaign Sanitary District,
as quoted by Brown (1937), mentions that passage of snails from the
sprinkling filters into the secondary settling tank, . .proves beyond
doubt the presence of a high amount of dissolved oxygen in the lower
part of the filters. " Associated with this quotation is the statement by
Brown that Physa anatina breathes atmospheric oxygen. Referring to
other records of snails reported from sewage treatment plants Brown
mentions that Physa halei has been reported from a Fort Worth, Texas,
installation
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Lohmeyer (1955) has written about the occurrence of an unidentified
Physa in a high-rate trickling filter of the University of Florida's
sewage treatment plant at Gainesville. In March of 1956 snails from
this plant were sent to the writer; they were forwarded to W. J. Clench
of the Museum of Comparative Zoology of Harvard University, who
identified them as Physa cubensis, a species of wide distribution in
Florida and the West Indies,, Lohmeyer does not give operational
data relative to the character of water that is applied to the filter. He
mentions that dosing the filter for three days with a chlorine residual
of approximately 3 pLp.m. resulted in snail control for eight months
before operational difficulties were experienced. Mechanical
difficulties relating to high-rate filter operation, resulting from Physa
cubensis snails, are described in detail.
In May of 1955, individuals of varying sizes of Physa intergra were
collected by the writer from the rock beds of both standard and high-
rate trickling filters at the Dayton, Ohio, sewage treatment plant. Egg
masses were present on the undersides of stones in the top three inches
of the beds„ Such information would seem to indicate that this species
successfully carries on its life cycle in these trickling filter beds. The
following operational data represent extremes that were recorded for
water going onto the filters for two weeks preceding snail collections:
O. D. 59 to 131 p. p. m. ; total nitrogen 24. 4 to 240 8 p. p„ m„ ; ammonia
nitrogen 13 to 17. 9 p. p. , chlorides 122 to 128 p„ p. m„ , and D. O. ,
O. O p„ p„ m„ The dissolved oxygen in water leaving the filters varied
from 2. 7 to 4. 9 p. p. m. Hydrogen ion concentrations were not avail-
able for the stated period but for the month of April they were about 7. 1„
III. SOME STRUCTURAL AND LIFE CYCLE VARIATIONS RELATING TO
MOLLUSCAN SURVIVAL ABILITY IN ASSOCIATION WITH DOMESTIC
SEWAGE POLLUTION
Some structural and life cycle variations that relate to differing
molluscan abilities to survive septic conditions and a substrate of
sludge may consist of differences in: the type of respiratory organs,
ability to close the shell for extended periods, weight of shell, and life
cycle. Because of such differences among mollusks when associated
with domestic sewage pollution, certain of the lung-breathing snails
survive better than gill-breathers and certain fingernail clams are
more resistant than are the mussels.
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(1) Survival of Lung-Breathing Versus Gill-Breathing Snails
Snails possessing lungs can generally be expected to survive
under low dissolved oxygen or septic conditions because they
typically rely on atmospheric oxygen in breathing. It is important
for the aquatic biologist to become familiar with snail morphological
characteristics in order to separate readily gill-breathing from
air-breathing snails, An obvious character revealing whether
the gastropod being dealt with is a gill-breather or a lung-breather
is the presence of an operculum in the aperture of the former.
Families and common genera of gill-breathing fresh water snails
are listed in Table 4 for convenience in separating them from those
that typically breathe atmospheric oxygen.
Table 4
FAMILIES AND GENERA OF GILL-BREATHING SNAILS
Family
Genera
Amnicolidae*
Amnicola
Pyrgulopsis
Hydrobia
Somatogyrus
Lyogysus
Bulimus (Bithynia)
Pleuroceridae
Pleurocera
Goniobasis
Valvatidae
Valvata
Viviparidae
Vivipara
Gampeloma
* Amnicolidae genera listed from Berry, 1943;
other families and genera from Baker, 1902.
It is important to point out to those conducting investigations in
lakes or deep rivers that the presence of a lung in a snail does not
necessarily mean that atmospheric oxygen will serve as the only
source for respiration in all seasons. Periodic movement of lung-
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breathing snail, from shallow to deep water, may be induced by a
lowering of water temperature to 10°C. as fall merges into winter
and may be an annual occurrence as reported by Cheatum (1934)
in Douglas Lake, Michigan. A change in temperature from 10°C. to
21°C is accompanied by an ascent from deeper water to marginal
lake areas. Cheatum presents data indicating that when pulmonate
snails are submerged, the lung functions as a gill in taking dissolved
oxygen from water. Lung-breathing snails may remain submerged
for three or four months out of every twelve in areas where winter
months are climatically similar to Michigan.
Experimental work conducted by Cheatum indicates that lung snails
are not completely dormant when submerged, but rather are in a state
of suspended animation, and that they actively respire using dissolved
oxygen* Individuals of the following seven species and subspecies of
lung-breathing snails are noted by Cheatum to breathe dissolved
rather than atmospheric oxygen for a part of each year when submerged
in Douglas Lake, Michigan: Lymnaea stagnalis appressa, Lymnaea
emarginata angulata, Helisoma campanulatum, Helisoma smithii, Helisoma
antrosum percarinatum] Fhysa sayii crassa, Physa parkerTI ' ~
In submerged experimental cages individuals of each of the
species named above survived for 65 days with the water temperature
varying between 16. 8 to 26. 6 C. Submergence experiments indicat-
ed that mortality was greatest among individuals whose oxygen
requirements were the highest. The presented data does not indicate
that submerged pulmonate snails can live under conditions of septicity.
In writing about the completion of life cycles, by certain pulmonate
snails, when submerged Cheatum (1934) states, "In all probability,
many individuals of the species H. campanulatum smithii, H. antrosum
percarinatum, L. emarginata angulata, and P. sayii crassa complete
their life cycles and reproduce normally witKout emerging for air. 11
Shelford (1913) writes that pulmonate snails of the genus Ancylus,
". . . are said to take water into their lung and thus do not need to
come to the surface for air. "
It is especially pertinent to point out literature, like the above,
concerning lung-breathing snails that can obtain oxygen from either
the atmosphere or from water, because in deep water during sub-
merged living conditions in winter certain pulmonates would be killed
by oxygen-consuming pollutants that could lower oxygen to asphyxial levels.
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Thus, it may not be advisable to always include pulmonate snails with
sludge worms, certain blood worms, rat-tailed maggots, and house-
hold or sewage mosquitoes as being tolerant to pollutional conditions
involving septicity.
Of all snail genera, members of the genus Physa, especially,
may occur in great abundance in septic zones of streams. Two
species, Physa interga and Physa anatina, are commonly associated
with septic zones in shallow streams in the mid-west during summer
and fall months.
The writer has not collected any of the gill-breathing snails in
polluted water where the dissolved oxygen, as measured during day-
light hours, was less than 2 p„ p. m. Even though these mollusks
possess an operculum which, if tightly sealed, should enable them
to close themselves away from low dissolved oxygen waters, the
fact that such snails are not reported from septic or near septic water
would indicate that low dissolved oxygen may be one of several factors
denying such water to them.
(2) Survival Relating to Shell Closure in the Sphaeriidae and Unionidae
The fact that certain of the Sphaeriidae can survive low dissolved
oxygen conditions and a shifting bottom of sludge, as related to domestic
sewage, and that the Unionidae do not poirtssomewhat speculatively to
the ability of certain fingernail clams to close the shell and survive
until stream conditions improve.
Allen (1923), in studying reactions of certain Unionidae under low
dissolved oxygen conditions writes, "When under conditions of deficient
oxygen not only to the siphons widen to bring in more water, but also
additional spiaces between the mantle edges are thrown open. " He
does not mention whether all of the Unionidae that he studied opened
the valves as indicated under the stated circumstances. The following
Unionidae are listed as being used in general experiments: Anodonta
grandis, Lampsilis luteolus^ L. ligamentinus, L. altus, Quadrula
heros, Q. pustulosa, Q. lachrymesa, Q. undulata, Unio crassidens,
U. gibbosus, Plagiola elegans, "acri(i otKers. " If the (Jnionidae, in
general, have a response to open their valves under low dissolved
oxygen conditions resulting from pollution by domestic sewage and
industrial wastes, they are most vulnerable to destruction. If they
do open their valves as described by Allen, their bodies are vulner-
able to any number of substances in polluted water that may be toxic
enough to destroy them. Also, in an open position they could be
covered by settable solids.
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It is known that the Sphaeriidae can live under septic conditions
and on sludge-covered bottoms as discussed earlier. Apparently,
when living under conditions of septicity the valves remain tightly
closed. Thus, the Sphaeriidae would not be subjected to toxic
materials as they would if they lived under such conditions with
their valves opened. Juday (1908) has written about the behavior of
Corneocyclas [=Pisidium] idahoensis under laboratory conditions in
water containing and devoid of dissolved oxygen, and has related
such data to field conditions. In water without dissolved oxygen,
individuals remained quiescent with their valves tightly closed with-
out activity being observed in the mud of the experimental jars. When
individuals were placed in aerated water they became active. He
states that his experiments seemingly indicate that this mollusk remains
quiescent or dormant in Lake Mendota, Wisconsin, when the muddy ooze
at the bottom of the lake contains no dissolved oxygen, a period of
about three months each summer. Juday (1921), in further studying
Pisidium idahoensis in lake Mendota, writes that there is no free
oxygen below a depth of 20 meters from about the middle of July until
early October, and again in March for two or three weeks in some
years. He mentions that organisms living under such conditions
must be . . facultative anaerobes. ", and includes in this category,
in addition to Pisidium idahoensis, worms of the genera Tubifex
and Limnodrilus, and three dipterous larvae: Corethra punctipennis,
Chironomus tentans, and Protenthes choreus.
Baker (1928) writes about Sphaeriidae being able to live in the
mud bottom of pools where the water has dried up, and Ingram (1941)
has reported Pisidium abditum living out of water on the beach of a
lake from at least June 15 to September 1.
(3) Survival Relating to Weight of Shell
The entombment effect of heavy sludge or silt pollution may relate
to the absence of heavy Unionidae and presence of certain light
Sphaeridae, other factors being favorable. In Dawley's (1947)
study of the distribution of aquatic mollusks in Minnesota, he
comments on the survival of mollusks on varying substrates, "A
mussel [Unionidae] is more exacting in its requirements than a
snail, being heavier and less motile. The bottom in which it lives
may be sand, gravel, or mud, but not rock or soft muck because
its foot cannot penetrate rock and it sinks too far into the muck and
is smothered. " Based upon commonly finding certain Sphaeriidae on
a sludge bottom, such a physical substrate may not deter the existence
of certain species of this family. General observations, based upon
reconnaissance of flocculent bottoms in rivers and streams polluted
by domestic sewage, indicate that the Unionidae do not seem to favor
such areas.
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In studies of erosion silt as a pollutant under laboratory
conditions, Ellis (1936) found that certain mussels were unable
to maintain themselves, in either sand or gravel bottoms, when
layer of silt from one-fourth to an inch in depth was allowed to
accumulate over such, other conditions being favorable to
survival. The yellow sand-shell, Lampsilis teres, a sand species,
most readily succumbed; the species least readily killed were:
Obliquaria reflexa, Quadrula quadrula and Q„ metanevra.
Coker et. al. (1922) compiled data of various investigators on
types of bottoms on which mussels were reported to be living.
From his analysis of such data he concluded, "It appears that the
preferred bottom for the majority of species is mud (but not deep,
soft mud, to which type of bottom few species are adapted) and
gravel, including sand and gravel. Sand ranks next and sandy clay
last; but few species of mussels exhibit a preference for sand or
sandy clay, and only two are recorded (by one observer) as finding
the most favorable environment in a bottom of clay mixed with sand. "
Baker (1928), in writing about fresh-water clams of Wisconsin,
discusses types of bottoms that mussels prefer: gravel, sand, mud,
and clay; he says that they are common or abundant in the first three
and rare in the latter. A shifting bottom, whether it consists of mud
or sand, is stated to be usually devoid of mussels. Fine silt bottoms
are always avoided by mussels, and Baker (1928) doubts if mussels
could live in such a bottom environment. He states that mussels are
usually absent or rare where great quantities of silt are carried into
streams. Ingram (1948) reported Anodonta wahlamatensis by the
thousands in the soft mud bottom of Stow Lake, San Francisco,
California.
Many fresh-water snails are heavy enough to sink into the sludge
covering stream bottoms to become buried and suffocate. The
writer has observed areas of streams where sludge deposits were
two to three feet in thickness, such as reaches of the Mahoning below
Warren and Youngstown, Ohio, where Physa intergra used higher
aquatic plants as a substrate rather than the flocculent sludge deposits.
In sludge filled sections of streams without higher aquatic plants,
snails may be found on rock islands protruding from the sludge, and
may be absent or rarely occur on sludge. In weight, adult fresh-
water snails are much more comparable to the Sphaeriidae than to
the Unionidae.
(4) Survival Relating to Type of Life Cycle
Of clams, the Unionidae are especially vulnerable to pollution
which may eliminate species by affecting larval stages. It is well
known that after being released from the female the immature
glochidial stage of the Unionidae must parasitize various fish in
order to assure life cycle completion, Lefevre and Curtis (1912),
Coker et. al. (1922), van der Schalie (1938), and Jones (1950).
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After spending from 10 to 14 days as an external fish parasite,
the glochidium drops off the fish and continues its life as a free
living form. If a fish is not parashtized, the glochidium dies.
No information is available on the direct effects of pollution on
the glochidium or on sperm cells which pass freely in water
from male to female clam.
Because it is necessary that a part of any Unionid's life cycle
be spent as a fish parasite, there is a direct relationship between
the effects of pollution on fish and perpetuation of succeeding
generations of Unionidae in any stream. If adult Unionidae are
more resistant to various pollutional affects than fish, they may
survive to die of old age, without succeeding generations develop-
ing to replace them. If glochidia-carrying fish are denied areas
of streams by pollutants, expanded distribution of the Unionidae
is hindered. A number of fish have been reported in the literature
as carrying glochidia of various Unionidae, Coker et. ah (1922),
Danglade (1922), Murphy (1942), Ingram (1948), Jones (1950).
The following fish are examples of some that have been associated
with glochidia, and are noted so that those working in water
pollution might be aware of them if it is ever desired to correlate
mussel-fish relationships relative to pollution: black bullhead,
common bullhead, bowfin, eel, sheepshead, gizzard shad, mooneye,
pike, spotted catfish, yellow catfish, long and short-nosed gars,
red-ear sunfish, orange-spotted sunfish, blue-gill sunfish, small^
mouth black bass, largemouth black bass, striped bass, river
herring, yellow perch, white crappie, black crappie, sand sturgeon,
madtom, sauger, and drum.
The Sphaeriid's sex cells are not subjected to any possible
pollutional effect outside of the adult's body. They are hermaphro-
dites, fertilization is internal, and the young may be carried in
the adult for as long as a year, Goodrich and van der Schalie (1944).
The growth stage that leaves the parent to fend for itself is a
small mirror-image of the adult. Such protected reproduction and
shielding of the very young, when compared with the haphazard
early life cycle stages of the Unionidae, should enhance survival
of fingernail clams over mussels.
Gastropods that one would encounter in water pollution
investigations copulate with resulting internal fertilization. Most
lay eggs that are attached to submerged objects and, on occasion,
to each other's shells; the Viviparidae are ovoviviparous. Thus,
the eggs and very young stages of most are exposed to external
changing environmental conditions at all times.
IV. NATURAL VARIATIONS IN DISTRIBUTION OF MOLLUSKS NOT
RELATED TO POLLUTION
In studying the effects of pollution on bottom organisms, with
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emphasis on mollusks, cognizance should be taken of natural
phenomena affecting distribution not related to pollution.
Normal variations in kinds, sizey and abundance of mollusks,
unrelated to pollution, make inventories of species of little
value in pollution studies unless those interested in delineating
indicator organisms include chemical, physical, and bacterio-
logical descriptions of water quality so as to establish tolerances
of mollusks to pollutants.
It has been shown by Baker (1918) that in lakes the numbers of
molluscan species decrease with depth. In further writings about
the increase of mollusk abundance in relation to depth, with
reference only to mussels, Baker (1928) states that "The great
majority of naiades live in comparatively shallow water from a
foot to six feet in depth. More rarely they descend to depths as
great as 25 feet. Records of fresh water mussels from greater
depths than 25 feet are to be viewed with suspicion. 11 Thus, in
studying the effects of pollution on benthonic organisms in a lake,
one should always be aware that paucity of a variety of mollusks
may naturally be related to water depth and not to pollutional
effects. In such studies chemical, physical, and bacteriological
tests could be most important in presenting data to indicate whether
a reduction of molluscan variety was a natural phenomenon of
depth or whether it could be attributed to pollution.
In streams, it is known that Unionidae and Gastropods tend to
increase in numbers of species from headwaters to the stream mouth,
Goodrich and van der Schalie (1944), Baker (1928). For example,
Baker (1928) lists on increase of Unionidae from three species
upstream to 28 downstream in a 27 mile reach of the Big Vermilion
River, Illinois. Certain pollution sources on the headwaters of a
stream may be suspect in relation to a dearth of mollusks such as
the Unionidae; however, a small number of species may represent
a natural condition rather than a relationship to pollution.
There may be a greater number of species and individual gastropods
living in stream areas where higher aquatic plants are present and
usuable as a substrate in addition to the stream or lake bed. Thus,
it is important to select stations to include sampling of higher
aquatic plants in studies designed to provide data on indicator
organisms. For example, in certain reaches of the Mahoning River,
Ohio, in 1952, the writer made collections of bottom organisms
in sludge deposits two to three feet in thickness and found no
mollusks. In those reaches the river had a water temperature of
96 F. , pH of 4. 1, and D. O. of 0. 2 An initial conclusion from
these meager chemical and physical analyses could have been that
mollusks were unable to stand such conditions in this stream. How-
ever, Physa integra was present by the hundreds in various growth
stages as well as in egg masses, using higher aquatic plants as a
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substrate. Thus, some data were collected to show certain
conditions under which Physa integra can survive and carry out
its life cycle. If higher plants present had not been searched
for organisms, one might not have associated this snail at all
with such a low pH or high temperature. On the basis of
collections limited to the stream bottom, this pulmonate snail
would have been associated only with waters having more
favorable pH and temperature and with but little sludge.
V. IDENTIFICATION SOURCES FOR FRESH WATER MOLLUSKS
To assist those interested in the relationship of mollusks to
water pollution, certain publications which may serve as examples
of aids to their identification are cited. Also, certain museums
having collections available for comparison of species or personnel
that can assist in identification of specimens are presented. Much
additional information relative to identification can be obtained by
literature searches, or by consulting State and municipal museums
and natural history societies.
A great deal of information concerning fresh water mollusks is
contained in various numbers of "The Nautilus, " a quarterly
journal devoted to the interests of conchologists. This journal is
edited by Dr. H. B. Baker of the University of Pennsylvania's
Zoological laboratories, Philadelphia, Pennsylvania.
The foremost nuseums housing collections of fresh water
mollusks are: the United States National Museum, Washington,
D. C., with Drs. Harald Rehder as Curator of Mollusks and
Dr. J. P. E. Morrison as Associate Curator; the Academy of
Natural Sciences of Philadelphia, Pennsylvania, with Dr. Henry
A. Pilsbxy as Curator of Mollusks and Dr. R. Tucker Abbott as
Curator of the Pilsbry Chair of Malacology; Museum of Comparative
Zoology of Harvard University, Cambridge, Massachusetts, with
William J. Clench as Curator of Mollusks; Chicago Museum of
Natural History, Chicago, Illinois, with Dr. Fritz Haas as Curator
of Mollusks; Museum of Zoology, University of Michigan, Ann Arbor,
Michigan, with Dr. Henry van der Schalie as Curator of Mollusks;
California Academy of Sciences, San Francisco, California, with
Dr. G. Dallas Hanna as Curator of Mollusks and Dr. Leo George
Hertlein as Associate Curator; and the Carnegie Museum,
Pittsburgh, Pennsylvania. Smith's (1943) directory of malacologists
can be useful to those interested in having mollusks identified,
because it lists in alphabetical order, malacologists who are
specialists in mollusk identifications.
The following are cited as examples of keys and faunal liats
developed from studies limited geographically that can serve to
provide species names as a base for specific identification in future
studies of fresh water mollusca: Baker (1922) on mollusks of the
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Big Vermilion River, Illinois; Eddy and Hodson (1950) on
mollusks of the North Central states; Goodrich and van der
Schalie (1939) on mollusks of the upper peninsula of Michigan;
Morrison (1932) on mollusks of the northeastern Wisconsin lake
area; Strecker (1931) on the Unionidae of Texas; and a series
of papers by van der Schalie on the Unionidae bearing the follow-
ing dates from the stated geographical areas" (1936) St. Joseph
River, southwest Michigan; (1938b) Cahaba River in northern
Alabama and (1938) Huron River in southeastern Michigan; (1940)
Chipola River in northwestern Florida; (1948) commercially
valuable mussels of the Grand River, Michigan; and (1950) of
the Mississippi River.
Examples of regional fresh water mollusk identification guides
are: Baker (1898) on clams of the Chicago area and (1902) on
gastropods of the same area; Baker (1928) on clams and snails of
Wisconsin; Chamberlin and Jones (1929) on the Mollusca of Utah;
Goodrich (1932) on mollusca of Michigan; Goodrich and van der
Schalie (1944) on mollusca of Indiana; Henderson (1924) (1936) on
mollusca of Colorado, Utah, Montana, Idaho and Wyoming;
Henderson (1929 and 1936a) on mollusca of Oregon and Washington;
Ingram (1948) on the larger fresh water clams of California, Oregon
and Washington; and Strecker (1935) on the mollusca of Texas.
The following are monographs, papers, and catalogues of
varing degrees of comprehensiveness that deal with fresh water
molluscan classification: Baker (1911) on the Lymnaeidae of
North and Middle America, Berry (1943) on the Amnicolidae of
Michigan; Brooks and Herrington (1944) on a preliminary survey
of the Sphaeriidae; Ortman (1919)> a monograph of the naiades of
Pennsylvania; Pennak (1953), a key to the families and genera of
North American fresh water mollusks; Sterki (1916), a catalogue
of North American Sphaeriidae; and Walker (1918), a synopsis
of fresh water mollusca of North America north of Mexico and
(1918), keys to fresh water mollusca of the United States.
VI. SUMMARY
1. Presented literature affirms that little specific information
is available relating one species of mollusk to a specific or over-all
set of well-defined water quality conditions, and another species
to different degrees of water quality.
No information is available to be used in selecting from
mollusks any species that may be called a pollution indicator.
Several species are tolerant to low dissolved oxygen or septic
conditions resulting from pollution of water by domestic sewage.
However, such species cannot be considered to indicate pollution,
as related to septicity, for they are also found in umpolluted streams
and lakes.
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2. Most specific data available relating mollusks to certain
effects of pollution on water quality deal with survival under
septicity. Other such data relate to survival in relation to
blanketing bottom deposits and acid water.
a. In relation to survival under conditions of septicity
resulting from domestic sewage pollution, general conclusions
that have been apparent for many years, can be reaffirmed here:
certain Sphaeriidae are collected under such conditions while
Unionidae are not; and certain pulmonate snails are commonly
found in septic water, while gill-breathers typically are not.
b. In relation to survival on bottoms blanketed by sludge
from domestic sewage and by silt, certain Sphaeriidae and
Physidae are most often collected.
3. Information on hydrogen ion concentration is presented
indicating that a number of genera of mollusks may live naturally
in acid water conditions not associated with pollutants.
4. Variations in molluscan structure and types of life cycle
are discussed, as such variations may result in enhancing
survival under pollutional conditions relating to dissolved oxygen,
sludge and silt deposits.
5. Natural variations in the distribution of mollusks in
streams and lakes are considered in order to make those working
in water pollution aware that variations in numbers of species may
be a natural phenomenon not related to pollution.
6. Selected museums and personnel as well as mollusk
guides and lists are presented to assist those who are not authorities
on molluscan taxonomy with identification.
VII. ACKNOWLEDGMENTS
Appreciation is expressed to Dr. Henry van der Schalie,
Curator of Mollusks, University of Michigan, Ann Arbor, Michigan,
for providing certain bibliographical data and correspondence
relative to mollusks and pollutional relationships. Credit is due
Mr. William J. Clench, Curator of Mollusks, Harvard University,
Cambridge, Massachusetts, who identified Physa snails associated
with trickling filters. ""
Thanks are due to Drs. A. F. Bartsch, W. Bridge Cooke, C.
Mervin Palmer, and Clarence M. Tarzwell of Water Supply and Water
Pollution Control. Robert A. Taft. Sanitary Engineering Center,
U.S. Public Health Service, Cincinnati, Ohio, for suggestions that
have improved this paper .
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Goodrich, C. and van der Schalie, H.
1944. A revision of the Mollusca of Indiana.
American Midland Naturalist, Vol. 32 (2), pp. 257-326.
Goodrich, C.
1945. Goniobasis livescens of Michigan.
Miscellaneous Publications, Museum of Zoology,
University of Michigan, No. 64,
Ann Arbor, Michigan, pp. 1-36.
Henderson C.
1949. Value of the Bottom Sampler in Demonstrating the Effects of
Pollution on Fish-food Organisms and Fish in The Shenandoah
River.
Progressive Fish Culturist, Vol. 11 (4), pp. 217-230.
Henderson, J.
1924. Mollusca of Colorado, Utah, Montana, Idaho, and Wyoming.
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Henderson, J.
1929. Non-marine Mollusca of Oregon and Washington.
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Henderson, J.
1936. Mollusca of Colorado, Utah, Montana, Idaho, and Wyoming-
Supplement.
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Henderson, J.
1936a. The Non-marine Mollusca of Oregon and Washington-Supplemen
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Hoskins, J. K. , Ruchhoft, C. C. and Williams, L. G.
1927. A Study of the Pollution and Natural Purification of the
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Ingram, W. M.
1941. Survival of Freshwater Mollusks during Periods of Dryness.
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-129-
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Ingram, W. M.
1948. The Larger Freshwater Clams of California, Oregon, and
Washington.
Jour, of Entomology and, Zoology, Vol. 40 (4), pp. 72-92.
Ingram, W. M. , Ballinger, D. G. , and Gaufin, A. R.
1953. Relationship of Sphaerium solidulum Prime to Organic Pollution.
The Ohio Jour, of Science. Vol. LIII (4), pp. 230-235.
Jewell, M. E.
1920. The Quality of Water in the Sangamon River.
State of Illinois Dept. of Registration and Education,
State Water Survey, Urbana, Illinois
Bulletin No. 16 Chemical and Biological Survey of the Waters
of Illinois, Report for the Years 1918 and 1919. pp. 230-246.
Jewell, M.E.
1922. The Fauna of an Acid Stream.
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1950. Propagation of Freshwater Mussels.
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1908. Some Aquatic Invertebrates that Live Under Anaerobic Conditions.
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1921. Quantitative Studies of the Bottom Fauna in the Deep Waters of
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1955. Snails in the Trickling Filter.
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Morrison, J. P. E.
1932. A Report on the Mollusca of the Northeastern Wisconsin
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Trans. Wisconsin Academy of Sciences, Arts and Letters,
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1942. Relationships of the Freshwater Mussels to Trout in the
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131-
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Richardson, R. E.
1921. Changes in the Bottom and Shore Fauna of the Middle Illinois
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Strecker, J. K.
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1936. The Naiad Fauna of the St. Joseph River Drainage in Southwest*
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American Midland Naturalist, Vol. 17 (2), pp. 523-527.
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1936a. Contributing Factors in the Depletion of Naiades in Eastern
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1938a. Hitch-hiking Mussels and Pearl Buttons.
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van der Schalie, H.
1938b. The Naiades (Freshwater Mussels) of the Cahaba River in Nortl
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1940. The Naiad Fauna of the Chipola River in Northwestern Florida.
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van der Schalie, H.
1941. Zoogeography of Naiades in the Grand and Muskegon Rivers of
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van der Schalie, H.
1948. The Commercially Valuable Mussels of the Grand River in
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Michigan Dept. of Conservation, Miscellaneous Pub I, No 4
pp. 1-42. " '
van der Schalie, H. and A. van der Schalie
1950. The Mussels of the Mississippi River.
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Walker, B.
1918. A Synopsis of the Classification of the Freshwater Mollusca of
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1917. Studies on the Digestion of a Sewage-filter Effluent by a Small
and Otherwise Unpolluted Stream .
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Experiment Station, Vol. 10, pp. 1-63. ®
Whipple, G. C., Fair, G. M. and M. C. Whipple
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-1 34-
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Wiebe, A. H.
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pp. 137-167.
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THE USE AND VALUE UJb AUUAT1U WNijJi.U i'5
AS INDICATORS OF ORGANIC ENRICHMENT
Arden R. Gaufin*, University of Utah
Salt Lake City, Utah
A knowledge of the ecological requirements of aquatic organisms is
of outstanding importance in judging the extent of pollution due to organic
enrichment in our streams. The species composition of the aquatic popula-
tion in a given area is determined by the environmental conditions which
have prevailed during the developmental period of the organisms involved.
If at any time during its development, environmental conditions become lethal
for a given organism, that organism will be eliminated even though the un-
favorable conditions are of very short duration. The aquatic population
which occurs in a given area is, therefore, a representation or indicator
of environmental conditions which have prevailed during the life history
of the organisms comprising the population.
It is this property of indicating past environmental conditions, especially
the extreme conditions of brief duration, that make macro-invertebrates
such valuable indicators of pollution. Most representatives of the group
have longer life histories than the micro-benthic fauna and are thus better
fitted for indicating past conditions than are the latter organisms . In addi-
tion, the larger siise and the mo'i-e distinctive morphological characteristics of
the macro-invertebrates make them easier to identify under field conditions .
The interpretation of stream conditions based on the biota present
has been used for many years . Kolkwitz and Marsson (1908-1909) first
proposed the use of aquatic organisms as indicators of the ecological
conditions under which they exist. They classified organisms as oligosa-
probic , mesosaprobic , and polysaprobic , depending on the concentration
of decomposable organic matter in the streams under consideration. Richards
(1921, 1928), based upon his studies of the Illinois River, deyeloped a classi-
fication of bottom organisms using seven groups of species. These included
a pollutional group, three sub-pollutional groups, an air breathing group,
current loving species other than pulmonate snails and air breathing insects
and clean water species. Patrick (1949) presented a comprehensive method
involving the use of histograms to show the response of certain groups of
aquatic organisms to given environmental conditions. This method was
modified and made more applicable to macro-invertebrates by Wurtz (1955).
A number of writers have published shorter works containing many
valuable ideas and suggestions. Bartsch (1948) presented information
on the response of the biota to various degrees of pollution. Gaufin and
Tarzwell (1952) employed both indicator species and associations in utilizing
aquatic invertebrates as indicators of pollution. Schiffman (1953) presented
a very useful method of cataloging stream bottom organisms found in,Illinois
~Formerly with the Aquatic Biology Unit of the Robert A. Taft Sanitary
Engineering Center, Cincinnati, Ohio
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with respect to their pollutional tolerance. Beck (1954) contributed a valuable
survey method in his simplified ecological classification of aquatic inver-
tebrates found in Florida streams .
In satisfactorily using macro-invertebrates as indicators of organic
enrichment and its effects, certain criteria should always be kept in mind.
First, several factors other than the presence of a pollutant may limit the
distribution of certain species, as for example, the type of bottom, speed of
current, lack of certain nutrients, scouring floods, excessive turbidity,
and the flight range. Second, the mode of occurrence of the forms considered
is just as important as their presence or absence in a given area. For ex-
ample, most of the macro-invertebrates which characteristically occur in
large numbers in heavily enriched water may also be found in limited numbers
in cleaner situations. Such forms as the mosquito, Culex pipiens, leech,
Macrobdella, and sludgeworms, Tubifex and Limnodrilus often occur in the
quieter confines of clean water streams in limited numbers, but they reach
far greater numbers in waters polluted by organic wastes. When conditions
are favorable for those organisms which can adapt to such conditions , they
thrive and build large populations. In some instances of organic pollution very
often the important factor in determining the occurrence of certain forms
is the abundant food supply which favors their growth and numerical increase
rather than a deficiency of some material such as dissolved oxygen. Similarly
those forms which are most characteristic of clean water conditions, such
as mayflies, stoneflies, or caddis flies are occasionally found during winter
in stream sections which are highly polluted or septic in summer.;' When
these insects drift into such a stream from nearby tributaries they may live
for considerable periods of time because the septic zone of summer often
has an adequate dissolved oxygen supply during winter. Thus, before such
isolated examples are taken as evidence that the forms involved
environmental conditions, an investigation as to their source and abundance
in the area is advisable.
In arriving at a better definition of the habitat preference and indicator
significance of the various groups of macro-invertebrates consideration of
the structural and physiological adaptations of the organisms is very im-
portant .
While there are exceptions, in general, an association of mayflies,
stoneflies, and caddis flies in a stream is indicative of clean water con-
ditions, and their absence often denotes a super abundance of organic wastes
and/or a low oxygen supply if the physical nature of the habitat is other-
wise suitable. Usually the presence or absence of' representatives of
other orders of aquatic insects which breathe by gills, and which are, there-
fore, dependent upon oxygen dissolved in the water for their respiratory
needs, has similar indicator significance. For example, while most aquatic
beetles can renew their oxygen supply directly from the atmosphere, a^nd
are thus unaffected by oxygen-depleting wastes, the larvae, pupae, and adults
of those species which are entirely aquatic, are dependent upon dissolved
oxygen and are restricted to clean water streams which are well aerated.
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In Ohio several species of riffle beetles such as Stenelmis crenata and
Stenelmis sexlineata, are found only in the cleanest streams, "their dis-
tribution in such a habitat indicates that the family, Elmidae, to which they
belong, is a member of the clean water association.
While most gill bearing aquatic insects are limited in their distribution
by low dissolved oxygen supplies, some forms which have more than one
toeans of respiration, such as the dragonflies and damselflies, display con-
siderable tolerance to low levels of dissolved oxygen. Their greater adapta-
bility to environments low in dissolved oxygen is made possible by the posses-
sion of respiratory structures which are the most highly developed of the
^arious gill systems. These insects can carry on respiration by means of
¦our different structures; namely, (1) caudal tracheal gills; (2) rectal folds;
) the integument; and (4) spiracles. Since all four organs may function at
he same time and many of the stream forms occur in either riffles or
'hallow marginal areas, the group is remarkably well adapted to withstand
He oxygen depleting effects of organic pollution. As a result of this adapta-
bility, the nymphs of both dragonflies and damselflies were often taken by
he author in Lytle Creek, Ohio, in sections of the recovery zone where the
dissolved oxygen supply in summer was as low as 1.0 p.p.m. for a short
ime during the night or in the early morning hours.
In streams receiving large amounts of organic wastes insects of the
'rders Hemiptera, Coleoptera, and Diptera have the most varied representa-
ion, are the most widespread in their distribution and are least affected by
dissolved oxygen concentration. Representatives of these orders may be
ound in all stream habitats representing all degrees of environmental
modification and stream recovery. Some species from each group may be
ound fairly widely distributed throughout the stream; others while not re-
tricted to either a clean water or "polluted" area, may show by their abun-
ance a strong preference for one or the other type of habitat. Still other
pecies, particularly among the Diptera, may be restricted to clean water
r to water rich in organic materials.
Of the three orders, the Hemiptera and Coleoptera are the poorest
indicators of organic enrichment and oxygen depletion in the stream. With
^e exception of the Elmidae, or riffle beetles, other species of beetles
nd all of the species of water bugs may be found throughout a stream
sually occurring most abundantly in the "polluted" areas where they may
¦nd an abundant food supply. The ability of members of these two orders
> withstand the oxygen-depleting effects of organic pollution is due to
pecial modifications of their tracheal system. These modifications serve
> increase the internal air capacity of the tracheal system, supplement
"acheal diffusion by ventilation movements when the insects come to the
urface for air, and provide supplementary external air stores . Common
> all of these forms are the modification of the body surface for break-
xg the water surface film, and changes in the wings and body surface for
apturing and holding stores of air, and in the tracheal system for sur-
ice ventilation and connection with the external air stores. In oxygen-
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deficient waters members of these two groups have only to increase the
frequency of their visits to the surface to cope with decreasing oxygen
supplies .
The efficiency of these modifications for obtaining and storing atmos-
pheric oxygen is well illustrated by Dytiscus , one of the diving beetles.
K, D. Roeder (1953) reports that this beetle can remain submerged for 36
hours without coming to the surface to renew its oxygen supply. Dytiscus ,
to obtain a supply of oxygen, breaks the surface periodically with hydrofuge
hairs, and ventilates violently by means of accessory respiratory muscles.
The fore wings, elytra, have a locking mechanism to trap the atmospheric
air, and the abdominal and thoracic spiracles are displaced so as to open
into this respiratory air store.
Aquatic Diptera may be found in a stream in many different ecological
niches in both the clean water and other life zones. However, with the
exception of only a few species, representatives of this order are highly
selective in their choice of habitat. A number of species such as Diamesa
nivoriundas Cricotopus absurdus , and Calopsectra neoflavella, are found
onTy in the cleanest;, most highly aerated sections of a stream, while others
such as the mosquito, Culex pipiens, and rattail maggot, Eristalis bastardi;
while found in limited numbers in clean water areas, show a decided'pre-
ference for the organically enriched sections . The variability in their choi<
of habitat and in their range of distribution is determined largely by the foo
getting and respiratory requirements and adaptations of the different indi-
vidual species- The larvae and pupae of the mosquito and rattail maggot,
with their special respiratory tubes, are unaffected by low oxygen supplies
as evidenced by the extremely large number of each that may be taken in
the most septic areas. Certain redblooded Chironomids, such as Chironom
ripariuSj also demonstrate a remarkable ability to thrive in the septic anc
recovery zones. Walshe (1950) has shown that the hemoglobin possessed by
midge larvae such as Chironomus riparius , Chironomus plumosus, and clos
related species apparently acts in both the transportation and storage of
oxygen. Its greatest transport role is during anaerobiosis» when it permits
the larva to continue filter feeding in low oxygen tensions and thereby in-
creases the rate of recovery from exposure to such conditions.
As in the case of the insects, the other groups of macro-invertebrates
show considerable variation in their distribution and adaptability to varying
environmental conditions due to the breakdown of organic matter. Certain
groups such as the slidge worms, Tubificidae, may be found in very large
numbers in bottom sludges of high organic content in the lower end of the
septic zone and the upper end of the recovery zone. Their numbers decreas
rapidly as the nature of the bottom sediments change. The ability of two ge
of these worms, Tubifex and Limnodrilus, to utilize the rich supply of orga
material under practically anaerobic conditions , make them important and
conspicuous members of one of the most easily recognized communities
characteristic of streams receiving organic wastes.
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Summary
In order to utilize macro-invertebrates as indicators of environmental
conditions in streamy it is essential to have a knowledge of the species compo-
sition and abundance of the various organisms in the populations involved,
under the various ecologic conditions which prevail in clean and organically-
enriched waters. Clean waters, with some exceptions, are characterized by
a great variety of invertebrates consisting of herbivores, carnivores,
and omnivores , prey and predators, lung, tracheal tube, and gill breathers.
In general apopulation containing numerous gill breathing forms as may-
flies, stoneflies , and caddis flies is indicative of clean water conditions and
their absence denotes the super abundence of organic materials and/or low
oxygen.
By contrast, associations engaged in the utilization of excess organic
materials are characterized by few species but large numbers of individ-
uals . The association of organisms normally present under the most septic
conditions consists largely of scavengers with few plant and animal eaters.
The macro-invertebrates most characteristic of septic zones are those
which can exist under conditions of very low oxygen or have adaptations for
breathing atmospheric oxygen.
Thus by reference to the qualitative and quantitative composition of
an aquatic population as an index of water quality, it is possible to delineate
the life zones in a polluted stream.
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Annotated Bibliography
Bartsch, A. F.
1948 Biological Aspects of Stream Pollution. Sew. Works Journ.,
20:292-302
Presents general ideas on the biotic response to various degree
of pollution.
Bartsch, A. F., and W. S. Churchill
1949 Biotic Responses to Stream Pollution During Artificial Stream
Reaeration. Limnological Aspects of Water Supply and Waste
Disposal. Publ. AAAS. pp . 33-48.
Graphically present the biotic responses which occur in the
various life zones of a stream receiving both sewage and
industrial wastes.
Beck, W. M., Jr .
1954 Studies in Stream Pollution Biology. I. A Simplified Ecolo-
gical Classification of Organisms. Quart. Journ. Fla , Acad.
Sci., 17(4):2 11-227 .
Develops a stream survey method in which macroscopic
invertebrates are used as biological indicators of ecological
conditions in Florida streams .
Gaufin, A. R. and C. M. Tarzwell
1952 Aquatic Invertebrates as Indicators of Stream Pollution.
Public Health Rep., 67:57-64.
Employ both indicator species and associations in utilizing
aquatic invertebrates as indicators of pollution.
Henderson, Croswell
1949 Value of the Bottom Sampler in Demonstrating the Effects
of Pollution on Fish-food Organisms and Fish in the Shenan-
doah River. Prog. Fish-Cult., (Oct „):2 17-2 30 .
Demonstrates the value of the Surber sampler for determining
the effects of pollution on aquatic organisms .
Kolkwitz, R., and M. Marsson
1908 Oekologie die pflanzlichen Saprobien. Ber.d. Deut. Bot.
Gesell. 26a:505-5 19 .
1909 Oekologie der tierischen Saprobien. Int. Rev. d. ges,
Hydrobiologie u. Hydrographie . 2:126-152.
First to propose the use of aquatic organisms as indicators
of the ecological conditions under which they exist. Classify
organisms as oligosaprobic, mesosaprobic , and polysaprobic ,
depending on the concentration of decomposable matter in the
streams under consideration.
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Patrick, Ruth
1949 A Proposed Biological Measure of Stream Conditions, Based
on a Survey of the Conestoga Basin, Lancaster County,
Pennsylvania. Proc . Acad. Nat. Sci. Phila., 101:277-341
Presents a comprehensive method involving the use of histo-
grams to show the response of certain groups of aquatic
organisms to given environmental conditions .
Richardson, Robert E.
1921 The Small Bottom and Shore Fauna of the Middle and Lower
"Illinois River and its Connecting Lakes, Chillicothe to Graftoi
Its Valuation; Its Source of Food Supply and Its Relation to the
Fishery.
III. Nat. Hist. Survey Bull. 13:363-522.
1928 The Bottom Fauna of the Middle Illinois River, 1913-1925.
ibid., 17:387-475.
Presents a classification of bottom organisms using seven
groups of species . These include 4 pollutional groups, an air
breathing group, and clean water species.
Roeder, K. D.
1953. Insect Physiology. John Wiley and Sons, New York. 1100 pp.
Considers the morphological adaptations and habits of repre-
sentative aquatic insects which fit them to particular habitats.
Schiffman, R. H.
1953 A Method of Cataloging Stream Bottom Organisms in Respect
to their Pollutional Tolerance. Mimeo. Publ. 111. Dept. of
Public Health.8 pp.
Classifies stream bottom organisms into three groups, toler-
ant, various, and intolerant, based upon their occurrence in
Illinois streams.
Walshe, B. M.
1950 Hemoglobin Function, Chironomus (Diptera). Jour. Exptl.
Biol., 27:73.
Shows that the hemoglobin possessed by midge larvae such
as Chironomus plumosus and closely related species appan-
ently acts in both the transportation and storage of oxygen,
and that its greatest transport role is during anaerobiosis.
Weston, R. S. and C. E. Turner
1917 Studies on the Digestion of a Sewage-Filter Effluent by a ,
Small and Otherwise Unpolluted Stream. Mass. Inst. Tech.,
Sanitary Research Lab., and Sewage Exp. Sta., 10, pp. 1-43.
Reveal the extreme importance and energetic action of cer-
tain plants and animals in purifying a stream receiving only
sewage effluents. Contains, a list of indicator organisms
found in the stream.
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Whipple, G. C .
1948 The Microscopy of Drinking Water (Revised by G. M. Fair
and M. C. Whipple. 586 pp, John Wiley and Sons, N. Y.
Considers the processes and some typical organisms involved
in the self purification of streams. Contains an ecological
classification of such organisms.
Wilson, John N.
1953 Effect of Kraft Mill Wastes on Stream Bottom Fauna. Sew.
Ind. Wastes, 25(10):12 10-1218.
Shows the effect of Kraft mill wastes on the bottom fauna of
two western streams and considers the pollutional status of
such organisms.
Wurtz, C . B .
1955 Stream Biota and Stream Pollution. Sew. Ind. Wastes,
27(11): 1270-1278.
Presents a method for classifying the biota of streams and
rivers by means of histograms. Reduces all free living organis**1®
in such areas to five basic life-forms.
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BIOLOGICAL INDICES OF WATER POLLUTION,
WITH SPECIAL REFERENCE TO FISH POPULATIONS1
by
Peter Doudoroff
U. S. Public Health Service
and
Charles E. Warren
Department of Fish and Game Management
Ore^oA State College, Corvallis, Oregon
A number*of investigators have very recently published discussions
having to do with biological indices and biological measures of water
pollution (1) (2) (7) (13) (14) (15) (16) (26) (27) (28) (29) (30)
(36) (38). Fjer dings tad (12) has discussed some of the pertinent
European literature. The fundamental concepts presented by'these
authors are not original, for the idea that aquatic organisms can be useful
"indicators" of environmental conditions, and particularly of the degree
af pollution of water with organic wastes, has a long history (12).
Because of certain novel features and the relatively wide scope of the
Studies, and the broad implications of some of the conclusions, the work
af Patrick (26) (27) (28) (29) (30) has attracted much attention in the
Jnited States and seems to deserve the closest scrutiny.
Although much has been written about the various biological indices,
here has been no general agreement among the authors as to the meaning
>f some of the most important terms used in this literature and little
sffort to clarify the terminology. In view of the variety of backgrounds
tnd dominant interests of individuals concerned with waste disposal and
sdth the effects of wastes on receiving streams, it is not surprising that
he term "pollution" does not have exactly the same meaning for all. It
s regrettable that a variety of meanings have come to be associated with
echnical terms such as "biological indicator of pollution". Some of the
ifferences of opinion as to what the biological indices are and what may
e their utility doubtless stem from a lack of agreement on the meaning
f the word "pollution". Investigators proposing the use of different
idicators of pollution should have clarified, it would segm, their ideas
s to just what constitutes pollution, or, in other words, exactly what
Miscellaneous Paper No. 31, Oregon Agricultural Experiment Station.
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it is that the indicators can be expected to indicate. Too often this
has not been done, or the ideas and definitions presented have not been
carefully developed and appear to be unsound from a practical stand-
point.
Should the mere change (physicals chemical, or biological) of
some aquatic environment resulting from waste disposal be regarded
as pollution even when ordinary human use and enjoyment of the water
and of associated natural resources have not been affected adversely?
When there is evidence of environmental change, is this always reliable
evidence of damage to a valuable natural resource? May not certain
beneficial uses of water be sometimes seriously interfered with by the
introduction of wastes which may cause little or no detectable alteration
of biological communities? Have there been any studies which have
conclusively demonstrated a useable fixed relation between the biological
indices of pollution and the actual fate or change in value of aquatic
resources which are subject to damage by pollution? If water pollution
can be the result of introduction of any of a great variety of substances,
organic and inorganic, is it proper to refer to those biotic responses which
are only known to occur in the presence of putrescible organic wastes
(i.e. to organic enrichment of water) as "indices of pollution"? Can there
be any general biological solution for all problems of detection and
measurement of water pollution, or is effort being wasted in a search
for such a general solution? Are broad limnological investigations
being undertaken where intensive study and appraisal of supposedly
damaged natural resources of obvious value to man would be more pro-
fitable? Is immediate practical value of research results being claimed
improperly in an effort to justify fundamental limnological studies for
which no such justification should be necessary? These are questions
which all biologists interested in water pollution should perhaps ask
themselves. Many of these questions have no categorical answer, but
it is hoped that the following discussion will prove thought-provoking.
It may not only call attention to cer.tain inconsistencies in claims made
and terminology used, but may also indicate the need for revision of
objectives or a change of emphasis in pertinent future investigations.
Biological investigation now is an integral part of water pollution
detection and control, and biologists have become increasingly aware
of their opportunities for contributing to progress in this field of work.
Their ideas have been solicited and have been well received by other
specialists. In trying to aid the advancement of their science, biologists
owe it to their profession to seek thorough understanding of the practical
problems of water pollution control. Understanding the complexity of
these problems will make apparent the need for thorough and critical
testing of new ideas previous to their widespread practical application.
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First, it is necessary to consider the meaning of the term
"pollution". The introduction of any foreign substance which
merely alters the natural quality of water without materially inter-
fering with any likely use of the water cannot be said in a practical
sense to constitute pollution. Virtually every stream and lake in
any inhabited region, receives at least a trace of something which
measurably or not measurably alters the natural quality of the
water. What is significant or important from a practical standpoint
is not the mere presence of the added material, but its influence upon
the economic and esthetic value of the water, or on human welfare in
a broad sense. It appears that most authorities in the field of water
pollution control and abatement agree in defining water pollution as
an impairment of the suitability of water for any beneficial human use,
actual or potential, by any foreign material added thereto.
This definition agrees with repeatedly expressed judicial opinion,
that is, with definitions of "pollution" and of "clean water" established
by courts of law. The following legal definition, cited on page 100 of
"Water Quality Criteria", a publication of the California State Water
Pollution Control Board (4) is typical: "For the purposes of this
case, the word 'pollution1 means an impairment, with attendant injury,
to the use of water that plaintiffs are entitled to make. Unless the
introduction of extraneous matter so unfavorably affects such use, the
condition created is short of pollution. In reality, the thing forbidden
is the injury. The quantity introduced is immaterial. " Other definitions
cited agree essentially with this one.
In accordance with the above definition of the word pollution, a
iemonstrable change of some components of the biota of a stream clearly
caused by the discharge of some waste into the water is not invariably
5vidence of pollution, any more than is a demonstrable chemical change,
[f it cannot be reasonably asserted that a hazard to human health or
nterference with some beneficial use of the stream, such as fishing,
nust accompany a particular alteration of the biota, the change cannot
:orrectly be said to indicate pollution. Even the discharge of a waste
vhich eliminates virtually all organisms initially present in a very
imall or temporary stream capable of supporting no aquatic life of any
ralue to man is not necessarily pollution. Oxygen-depleting organic wasti
nay be thoroughly mineralized in such streams through natural self-
>urification processes, so that only harmless substances and"
leneficial plant nutrients may reach larger watercourses to which these
itrearns are tributary.
In agreement with the definition offered above, Beck (1) has defined
>ollution broadly as "the alteration of any body of water, by man, to such
degree that said body of water loses any of its value as a natural
esource. 11
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Patrick (28), on the other hand, has proposed a distinctly different,
strictly biological definition. This author defines pollution as "any thing
which brings about a reduction in the diversity of aquatic life and eventually
destroys the balance of life in a stream. " By way of explanation, it is
further stated that "As conservationists interested in using rivers today
but not abusing them so that they are damaged in the future - this is the
basis on which pollution should be judged. For it is by preserving the
biodynamic cycle that the ability of a river to rejuvenate itself is maintained.
Unfortunately it is not clear just what is to be regarded as pollution
according to the definition given by Patrick. Is any reduction in the
diversity of aquatic life evidence of pollution which will eventually destroy
the "balance of life", or only such a severe reduction of the diversity of
life that the ability of the stream to "rejuvenate itself" is indeed destroyed?
A reduction of species numbers is not always necessarily followed by the
eventual destruction of the "balance of life" in a stream and of the ability
of the stream to "rejuvenate itself" (i. e. » to undergo natural self-purificatic
Patrick (28) has pointed out that the so-called "food chain" in aquatic
environments "consists of many series of interlocking links so that if one
series is broken another can take over so that the chain is not destroyed. "
It is well known, also, that in certain "zones" of streams heavily and
continually enriched with organic wastes relatively few animal and plant
species are present, as a rule, yet natural purification proceeds at a
very rapid rate. Here, as in an efficient trickling filter, an ideally adapted
and obviously vigorous, healthy, and in certain respects very well balanced
biota of limited variety can exist, and the organic waste is mineralized far
more rapidly and efficiently than it could possibly be in a previously uncon-
taminated stream with its original, primitive biota. The ability of the
stream to "rejuvenate itself" certainly cannot be said to have been destroyed,
or even impaired.
Thus, a stream can be seriously polluted, in any usual sense of the
word, without lasting destruction of the "balance of life" and of self-
purification capacity (which balance hardly can be permanent anyway,
in any unstable environment). On the other hand, mere reduction of the
diversity of aquatic life without impairment of any important "food chain"
(i. e. , the food supply of valuable fishes, etc. ), or interference with
existing stream uses, does not necessarily have anything to do with the
conservation of natural resources. It appears,'therefore, that the last-
mentioned definition of pollution is unsatisfactory, from a practical stand-
point, no matter how it was meant to be interpreted.
Careful consideration of the other pertinent writings of Patrick and
of the proposed method oi judging stream conditions leads to the con-
clusion that probably this author regards any marked reduction of the
diversity of aquatic life as evidence of pollution.
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Beck (1) states that "Patrick's methods suggest that the bio-
dynamic cycle should be maintained in the primitive condition, "
allowing for no equitable stream use, for "any deviation the
primitive bio-dynamic cycle is interpreted by Patrick as evidence
of pollution. " Actually Patrick has not suggested that an entirely
primitive condition of every stream biota should be maintained and
has classified as "healthy" certain stream sections which evidently
were not in the primitive state. A diversity of organisms approach-
ing that found under undisturbed or primitive conditions does seem
to have been regarded, however, as being characteristic of all "healthy",
unpolluted waters. This interpretation of Patrick's views may be right
or wrong. In any case, the need for clarification thereof, and for
better agreement among biologists as to the meaning of terms too often
loosely used, is apparent. It is noteworthy that Patrick's definition
of pollution, quoted above, implies that an alteration of water quality
cannot be pollution if it has no appreciable effect on the diversity of
aquatic life, and it can be interpreted as meaning that a marked reduction
of the diversity of aquatic life is always associated with pqllutional abuse
of the aquatic environment. Probably few if any workers directly
concerned with water pollution abatement or control can approve such
a definition.
One can hardly maintain that the relative worth of any biological
environment depends on the number of species that it supports, rather
than on the relative abundance of species of some importance or value
to man The presence of many different weeds does not usually contribute
to the value of a pasture. Also, it is not always correct to assume that
any marked modification of a natural environment and of its original,
primitive biota will result in their economic degradation, that is, a
reduction in value. The clearing, irrigation, and cultivation of desert
and other almost worthless lands, the application of agricultural and
other poisons for the control of various pests and weeds, and many
other human activities can, indeed, greatly enhance the value of the
affected lands while drastically modifying their biotas and reducing the
numbers of species present. Not only the production of valuable crops
is thus promoted, but sometimes also the production of equally valuable
wild game. On the other hand, the destruction of only one or a few
animal or plant species of outstanding value (e.g. , by some selective
poison) obviously can mean great loss. This loss is in no way
ameliorated by the fact that most of the organisms in the same environ-
ment are not noticeably affected. It is evident that a change of any
biota considered as a whole (e.g., the number of species represented,
etc. ) may not be a direct nor always reliable index and measure of
damage to any valuable natural resource. There seems to be no sound
basis for a general assumption of their strict or even approximate
parallelism.
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Although most authors evidently have recognized the economic
significance of pollution, it appears that when devising their
biological indices and measures of water pollution and its severity-
some biologists have completely disregarded all economic considerations.
They seem to have curiously attached at least as much importance to
the elimination of any species of diatom, protozoan, rotifer, or insect
as to the disappearance of the most valuable food.or game fish species.
Yet, some have claimed that their measure of the harmful effects of
pollution is a direct measure and therefore is more reliable than any
chemical evidence or measure of pollution. Why the fate of harmless
algal, protozoan or insect species can be said to indicate directly the
extent of damage to a valuable fish population or to any commercial,
recreational, or other use of water has not been explained.
If biological indices and measures of the severity of pollution can-
not be relied upon always to reveal even the extent of damage to valuable
aquatic life, they certainly do not indicate accurately the general
pollutional status of any water. Water which is rendered biologically
sterile by addition of some substances such as chlorine, or is appreciably
enriched with some organic wastes, other than domestic sewage, may be
of good sanitary quality and suitable for most ordinary domestic,
agricultural, and industrial uses. On the other hand, water in which
aquatic life is not markedly and adversely affected can be contaminated
with dangerous pathogens or with chemicals which may seriously
interfere with one or more of the above-mentioned uses. In view of
the great variety of water uses, and the number and complexity of
considerations (physical, chemical, biological, psychological, economic,
and sociological) which evidently must enter into any reliable determinatic
of the degree of interference with these uses by pollution, the evaluation
of the over-all pollutional damage cannot be a simple matter. Any
contention that some biological observations alone can cut across all
of this complexity and show clearly whether the actual and potential uses
of a stream have or have not been affected, and the magnitude of the
total damage, would appear to be an over-simplification of the problem.
It must be admitted that probably nobody has come forth yet with a clear
statement of this claim. And yet, unless a different meaning is made
perfectly clear, is not this claim implicit in every asseration to the
effect that a generally applicable and reliable biological index or measure
of the pollutional status or condition of streams has been devised and
developed?
Biotic responses to all of the numerous and very different water
pollutants are not alike. Early students of water pollution (23) (24)
(31) dealt chiefly with pollution by putrescible organic wastes and
particularly domestic sewage. In their day, the use of the term "biologic
indicators of pollution" when referring to organisms which respond in
a certain way to heavy organic enrichment of their medium was perhaps
justifiable. Untreated or inadequately treated domestic sewage then
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was by far the most important and perhaps the only well known and
generally recognized water pollutant. Its discharge into public waters
in amounts sufficient to bring about appreciable biotic changes being'
usually a hazard to human health, it was and is almost always pollution in
any ordinary sense of the word. Today, the importance of pollutants
other than domestic sewage is generally recognized. Yet, many
authors still speak of "pollution indicators" when they actually are
referring only to indicators of organic enrichment of water with
putrescible organic wastes, which may or may not involve demonstrable
damage to natural resources. Some readers are known to have been
misled by this terminology, believing that the same biological indices
are useful in detecting every kind of pollution.
Gaufin and Tarzwell (13), when reporting their studies of stream
pollution with domestic sewage, obviously were considering the effects
on aquatic life of an oxygen-depleting organic waste only. Nevertheless,
such unqualified and seemingly general statements as their conclusion
that "Pollutional associations are characterized by few species but
large numbers of individuals" can be misleading. As the quoted authors
well know, the numbers of many organisms initially present are reduced
and the numbers of none are markedly increased in some waters polluted
with toxic wastes, suspended solids such as silt, or even oxygen-depleting
organic wastes discharged intermittently. These authors undoubtedly
did not intend the conclusion in question to be a very broad generalization
from their observational results having to do with one kind of pollution
only. Their use of the expression "pollutional associations" for designating
associations found in waters polluted with domestic sewage, or in waters
enriched with putrescible organic matter, can be excused on the ground
that no term that is more appropriate than the term "pollutional" has come
into general use in the biological literature. Yet, this lack of a more precist
terminology is not any less deplorable because the use of inappropriate
terms, and terms which are not sufficiently specific, has become prevalent.
Beck (1) (2) explicitly confines his discussion to the subject of
"organic pollution". He has proposed the use of a numerical "biotic index",
which is said to be "indicative of the cleanliness (with regard to organic
pollution) of a portion of a stream or lake" (2). He recognizes that his
methods are "confined to fresh waters and encroaching salinity has a
marked effect on the fauna of a stream. " Inasmuch as many different
pollutants, including toxic constituents of some organic wasteB, likewise
can have a marked effect on the fauna of a stream, it is apparent that
Beck's methods may have only very limited applicability. It may be us-
able only in connection with the investigation and description of waters
known in advance to contain no pollutants other than non-toxic putrescible
organic matter.
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Patrick (26) (27) (28), recognizing the importance of a variety
of pollutants, apparently has attempted to devise a general procedure
for the reliable biological detection and measurement of the different
kinds of pollution. For reasons already indicated, however, this desir-
able objective appears to be attainable only when one defines pollution
as "any thing which brings about a reduction in the diversity of aquatic
life", which is not a generally acceptable definition.
Wurtz (38), while evidently realizing the existence and importance
of a large variety of pollutants, seems to overlook completely the
important differences of biotic responses to the different pollutants.
Thus., his Figure 1 suggests that the same pollutional. zones, including a
"degradation zone" extending from the point of mixing of an effluent
with the water of a stream to a "polluted zone" located some distance
downstream, can be expected to occur in any heavily polluted stream,
regardless of the nature of the pollutant (i. e. , whether it be "organic",
"toxic", or "physical"). Furthermore, he speaks of "pollution tolerant
species" and of "non-tolerant organisms", suggesting that organisms
are consistentely tolerant or consistently non-tolerant with respect to
all pollutants. Nowhere does he specify that he has in mind resistance
to putrescible organic pollutants only, and there is considerable evidence
that he has in mind all pollutants. In large degree, Wurtz seems to have
adopted methods similar to Patrick's, but one of his innovations seems
to require the probably impossible classification of all or nearly all
aquatic organisms as "tolerant" and "non-tolerant" to all kinds of
pollution, including the various toxicants, etc. Unfortunately, Wurtz
does not include in his paper a list of all organisms considered by him
to be tolerant and all those thought to be non-tolerant.
There can be no doubt that some of the so-called "pollution-tolerant"
organisms, which actually are simply forms known to thrive in waters
markedly enriched with organic wastes, are less tolerant with respect
to some other water pollutants than a number of the species known as
"clean-water" forms. For example, a species of Physa, a genus of
snails generally believed to be resistant to organic pollution (1) has
been found to be extremely susceptible to dissolved cqpper. Certain
fish (e.g., centrarchids), may fly nymphs, etc., thought to be more
susceptible than Physa to the effects of organic pollution, proved much
more resistant to copper. An aquatic environment in which "clean-
water" organisms are predominant might possibly be more seriously
polluted than one with decidedly "pollutional" biota. The biological
terminiology evidently needs revision, so that the word pollution would
not be used synonymously with organic enrichment.
It appears that, in general, very broad significance of the various
biological indices of water quality and the severity of pollution has been
only assumed and not actually demonstrated. This is well exemplified
by the following quotation from the summary of one of Patrick's papers
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(27): "On the premise that the balanced physiological activities of
aquatic life in surface waters are essential for the maintenance of
healthy water conditions, it may be assumed that the most direct
measure of this biodynamic cycle will indicate the condition of the
water. " It will be noted that we have here an assumption based upon
a rather nebulous premise. Most writers have failed to supply entirely
satisfactory, clear definitions of terms used (e. g. , "pollution",
"health", etc. ) to show precisely what it is that they believe they can
detect or measure biologically. Others have failed to use defined terms
in a manner entirely consistent with their own definitions. The need
for demonstration of the validity of some of the most fundamental assump-
tions concerning the reliability of pollution indices designed for general
application has not been satisfied. Some authors seem to be of the opinion
that the proof is unnecessary. It must be admitted that investigations
designed to provide such proof would be extremely complex and difficult,
and it is not likely that the search for this proof would be very rewarding,
for there can hardly be a simple, general solution for the problem of
pollution detection and measurement. Like a panacea, a general test
for all kinds of pollutional damage is something for which biologists and
engineers alike probably would be wise not to seek.
The value of fish as indicators of environment conditions and the
importance of fish population studies in connection with the estimation
af the intensity of water pollution now can be considered. Doubtless
ihere is much more published information on the environmental require-
ments of fish than on the requirements of species of any other group
af aquatic organisms excepting perhaps a few invertebrate species of
outstanding economic importance. The vast quantity of published data
relating to the water quality requirements of fish is partly revealed by
a. few recently prepared compilations - and summaries of some of this
nformation (4) (5) (8) (9) (10) (11) (17) (33). The resistance of
nany fish species to extreme temperatures, to unusual concentrations
>f dissolved oxygen and other dissolved gases, to variations of water
salinity, and to extremes of pH, their susceptibility to the harmful
sffects of a great variety of toxic substances and of suspended solids of
importance as water pollutants, the influence of some of these environ-
nental factors upon embryonic development, growth, and activity, and
to forth, have all been studied intensively. There exists also a voluminous
iterature on the food of fishes, their life history and reproductive
¦equirements, their habitat preferences, movements, avoidance of
.dverse environmental conditions, and so on.
While it is evident that more is known of the environmental require-
aents of many fish than is known of the requirements of most, if not
11, of the other aquatic organisms often considered as indicators of
nvironmental conditions, the use of fish as indicators has received
onsiderably less attention than has the use of other major groups, plant
nd animal, microscopic and macroscopic. Fisheries workers recognize
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the difficulty of adequately sampling fish populations even in bodies of
water of moderate size, and this, along with the mobility of fishes,
has been advanced as a reason for the unsuitability of fish as indicators
of environmental conditions. But, other aquatic groups are difficult
to sample too, as Needham and Usinger (25) have demonstrated in the
case of the invertebrate macrofauna of a riffle. The difficulty of samplin
and the mobility of fishes may not be the chief reasons why fish have
not been given more consideration as indicators. The taxonomic groups
which have received the most attention no doubt have reflected to some
extent the special interests of investigators who happened to be working
in the field of water pollution. Fish being the usual economic and
recreational yield of stream productivity, their study has obvious applied
value and so has required no additional justification. Further, the status
of a fish population may indicate suitable or unsuitable environmental
conditions, but when knowledge of this population is the end or aim of
an investigation, the population status is not regarded as an index of
anything else. The value of fish as indicators of the suitability of water f<
uses other than fishing has not been clearly demonstrated. Whatever the
reasons may be, the emphasis in most discussions of the "biological
indices" has been on groups other than fish, even though very little is
known of the environmental requirements of the species of many of these
groups.
The value of knowledge of fish populations in connection with the
classification of aquatic environments has not been entirely overlooked.
Ricker (32) made important use of the brook trout (Salvelinus fontinalis)
and the Centrarchidae and Esocidae as a basis for his ecological classift-
cation of certain Ontario streams. Fisheries workers frequently use suci
expressions as "trout waters" or "bass waters", thus conveniently
classifying waters according to the fish species for which the waters are
well suited. European workers have made more formal use of such a
system of stream classification (34) (37). Brinley and Katzin (3) have
classified waters and named various pollutional "zones" of streams in the
Ohio River drainage basin according to the kinds of fish populations found
therein. As has been done with other animals and plants, some species
of fish have been classified as to their "saprobic" preferences by a few
authors (22) (24) (19) (35), The basis for such classification of fish is
highly questionable. Patrick (26) (27) includes fish among the groups
considered in her "biological measure" of stream conditions. Doudoroff
(7) and Gaufin and Tarzwell (14) have emphasized the need for thorough
fish population studies in connection with water pollution investigations
and the determination of the pollutional status of waters.
Studies of fish populations in variously polluted waters, which reveal
varying susceptibility of different fish species to pollutional conditions
in their natural habitats, have been reported by a number of investigators
(3) (6) (11) (20). However, sufficiently intensive sampling of fish
populations has not often been undertaken in connection with routine polluti
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surveys and investigations, the sampling of other aquatic life having
been probably more often emphasized when the scope of the biological
studies has had to be limited. Inasmuch as it is not often possible
adequately to study all of the aquatic biota, including the fish, the practical
value of information to be obtained by concentrating attention on fish
populations must be carefully weighed against that of information to be
derived from equally intensive study of some of the other aquatic organisms,
and from comparatively superficial study of the entire biota.
The absence or extreme scarcity of some fish in a stream below the
ioint of entry of a waste, and not above the point of entry, strongly suggests
hat the waste is somehow detrimental to these fish, if valuable good and game
ish species are among those believed to be adversely affected pollution is
ndicated. Neither the presence nor the absence of fish is a reliable indication
>f suitability or unsuitability of water for domestic, agricultural, and
ndustrial uses and for recreational uses other than fishing. Nevertheless,
lecause of the great economic and recreational value of many fish species,
his information is essential to sound classification of waters according to
heir pollutional status.
The presence of fish does not necessarily show that their environment
as been suitable for them for a very long time, nor that the species found
an survive indefinitely and complete their life cycles under the existing
nvironmental conditions. However, the presence of thriving populations
f non-migratory species, including numerous representatives of different
ge classes whose growth rates have not been subnormal, is significant.
suggests strongly that pollution which is highly detrimental to these fishes
nd to migratory species whose habitat preferences, natural food, and
ater quality requirements are quite similar has not occurred recently,
ar example, the presence of numerous cottids in Northwestern salmon
id trout streams which receive organic wastes is believed to indicate that
ssolved oxygen concentrations have been adequate for some time and
:her environmental conditions probably have been suitable not only for
e cottids, but also for migratory salmon and trout. There is now no
>und reason for believing that the presence of any invertebrate form is
more reliable and appropriate biological indicator of the suitability of
ist environmental conditions for the migratory salmonids than is the
•esence of cottids.
The value of waters used for fishing, and of the fisheries which they
pport, bears no fixed, direct relation to the number of fish species to
found therein, just as it bears no such relation to the number of species
other organisms present. Some 35 species of fish were collected in the
Ldwestern warm-water stream studied by Katz and Gaufin (20). Because
the scarcity of valuable food and game fishes, this small, polluted
ream is not regarded as a valuable fishing stream. On the other hand.,
my cool, pure streams which are highly valued as trout and salmon
reams contain very few fish species other than the salmonids.
leed, the invasion of valuable trout waters by other fish
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species not initially present is generally regarded as evidence of degradation
of these waters, for the numbers of trout usually decline when it occurs.
Such a change of the fish population can be a result of increasing temperatures
and probably also of enrichment (18), Warrrv eutrophic waters can
support a great variety of fish and other organisms, but trout waters
which are approaching this condition can hardly be regarded as "healthy".
Some of the above statements seem to contradict Patrick's (26)
(27) conclusion, based on a study of the Conestoga River Basin of
Pennsylvania, that "The results of this study indicate that under healthy
conditions a great many species representing the various taxonomic groups
should be present. " It is necessary, therefore, to examine the evidence
on which the latter conclusion is based. It appears that, in accordance
with Patrick's conception of what a "healthy" stream should be like biologicall
only those stations where a variety of organisms judged to be fairly normal
or typical was actually found were classed as "healthy". It is not surprising*
therefore, that all of the stations classed as "healthy" had indeed this large
variety of organisms. Chemical, bacteriological, and other data were
collected and considered in selecting and classifying the stations studied.
It is clearly indicated, however, that the variety of organisms found (which
is the proposed index or measure of stream "health") also was a major
consideration. Different conclusions perhaps would have been reached had
the initial classification of the stations been based entirely on other criteria
of obvious practical import (such as the abundance,condition, and growth rates
of valuable native game fish, etc. ) and had a greater variety of natural,
unpolluted streams been examined, It is noteworthy also that certain stations
which evidently were not much affected by waste discharges but lacked the
usual variety of organisms (e.g., Station No. 152, in a stream section
evidently suited for stocking with trout) were classed as "atypical" stations
by reason of certain observed peculiarities, such as low water temperatures
unusual bottom or shore conditions, etc. Other stations which had the
expected variety of organisms were classified as "healthy" stations despite
noted peculiarities such as marked organic enrichment, unusually high BOD,
high CO2 content, high bacterial content, or great turbidity of the water.
Thus, it appears that the rating of the stations was somewhat arbitrary,
"When the possibility of certain pollutional damage specifically to
fisheries is under consideration, it should be remembered that fishes
have varying ecological requirements and habits, differ in their resistance
to variations of water quality, and are not all dependent upon all aquatic
organisms, nor upon the same organisms, for their food. It has been shown
that the growth of some fish species is promoted in certain waters affected
by the discharge of organic waste (21), whereas the same waters apparently
are rendered unsuitable for some other species (20). A reduction of the
number of species of fish-food organisms, with a great increase of abundance
of some of the remaining species, which occur often in streams receiving
various wastes, doubtless can be harmless or beneficial for some fish
species, although this reduction may be detrimental to others. If they are
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not otherwise adversely affected by environmental changes, those fishes
which can well utilize the abundant food organisms will thrive, while
others may disappear. Whether the total effect on fisheries will be
favorable or unfavorable clearly will depend on the relative commercial
and recreational value of those fish populations which are favored and
those which are affected adversely. An intensive study of the entire
aquatic biota cannot always reveal the extent of pollutional damage to
fisheries, unless the relative value of the various forms present (for man,
or as food for important fishes) is considered.
To evaluate the effect of environmental changes on fisheries it is
necessary to know what fish species were originally present, how highly
each is valued, and in what way and to what degree each important species
has been affected by waste discharges. The relative abundance and condition
of individuals of different species in the waters under investigation and in
suitable "control" areas, the growth-rates of different age classes, the
palatability of the flesh, and possible interference with normal migratory
movements or with other reproductive activities must all be considered.
Fish collection? taken by carefully planned netting will yield much of this
information. Commercial and sport catch records, showing the take per
unit of fishing effort, and various field observations (e.g. , of spawning
areas utilized, etc.) also can be very helpful. Inasmuch as the presence
of wastes and other pollutants is by no means the only factor which can
directly influence fish populations, the cause of observed differences of
fish populations must be determined. In this connection, studies of the
food of important fish species and of the relative abundance of available
food organisms in waters which are affected and those which are not
effected by waste discharges may be essential. However, if detection and
evaluation of pollutional damage to fisheries is the only or primary objective
af a biological investigation, an enumeration of the species of organisms
sf all taxonomic groups, or of some single invertebrate group, cannot be
ieemed a direct approach to the problem at hand. Judged only by its
iractical utility, it may be a waste of time, effort, and money, which
ierhaps could be far better expended on more directly pertinent studies,
indeed, it is difficult to imagine pollutional interference with any use or
:ombination of uses of water which could usually be accurately and most
sfficiently evaluated in such an indirect manner.
A study of the influence of large amounts of organic waste on the
scology of the Tuolumne River of California has recently been completed
>y Warren (unpublished data). During August and September of 1952,
he daily mean discharge rates of this river at the city of Modesto ranged
rom 293 to 822 cubic feet per second. The daily mean discharge rates
>f domestic and cannery waste introduced into the Tuolumne at Modesto
•anged from 0 to 22. 3 cubic feet per second. The 5-day biochemical
>xygen demand of samples of this waste ranged from 60 to 575 parts per
nillion. Dissolved oxygen concentrations at stations below the point of
vaste discharge ranged from zero to supersaturation during this time.
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The objective of this study was to determine some of the
effects of organic waste discharges on the ecology of the Tuolumne
during the different seasons of the year. Some thirty miles of the
river were studied, of which only the lower ten were influenced by
waste discharges. The phytoplankton, zooplankton, benthic fauna,
and fish were studied along with the physical, chemical, and bacteriological
conditions in this river. The fishery phase of the investigation represented
a Small part of the total effort.
The investigation of the Tuolumne River now being complete and its
objective more or less realized, it is interesting to consider how well
other objectives might have been satisfied by this same study, planned and
conducted as it was. For instance, had the objective been to determine the
influence of the_organic waste specifically on the fisheries of the Tuolumne,
could not much of the effort devoted to the bacteriological, phytoplankton,
zooplankton, and benthic faunal investigations have been far better expended
on a thorough study of the fisheries? One is forced to conclude that were
the objective to determine the status of the fisheries, the fish should have
received most of the attention. This does not mean that studies of the
plankton and of the benthic fauna are not necessary phases of an investigation
so oriented. They may be quite necessary, but they should be so planned
that the time and effort devoted thereto would not be out of proportion
to their contribution to thorough understanding of the status or condition
of the valuable fish populations.
The benthic fauna present at stations on the Tuolumne River below
the point of waste discharge had many of the recognized "pollutional"
characteristics during late summer and early fall. By this time, many
of the "clean-water" species present at these stations earlier in the
summer, and persisting at stations above the waste outfall, had disappeared*
A marked reduction in species numbers had taken place, and at least
one species occurred in unusually great numbers. While the bottom fauna
showed changes that in accordance with most biological index methods
would be regarded as evidence of pollution, rather intensive seining during
mid-September resulted in the collection of 10 species of fish at stations
above the point of waste discharge and 12 species at stations within the
first ten miles below this point. The variety of fish pfresent had certainly
not been greatly altered by the introduction of wastes, even though the
bottom fauna had been markedly modified.
Collections of young bluegills (Lepomis macrochirus ) made in
September showed the O-year class to grow faster at stations below the
point of waste introduction than at stations above this point. The size
difference persisted in the I-year class. The difference in the O-year -
class growth rates could probably be attributed to the greater abundance
of zooplankton at the downstream stations.
While the above data are interesting, they cannot be taken as evidence
that pollution of the Tuolumne damaging to fisheries did not exist. Some
evidence indicated interference with a portion of the upstream migration
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of adult Chinook salmon (Oncorhynchus tshawytscha), though the
downstream migrant young were presumably unaffected, being apparently
absent from the Tuolumne by the time of critical summer river flows
and waste discharges. Juvenile shad may perhaps have been affected
also. Had the principal objective of the Tuolumne River investigation
been an evaluation of damage to fisheries resources by pollution, the
study could not have been deemed complete in the absence of conclusive
evidence that interference with salmon migrations and other possible
damage to valuable fish populations had or had not occurred. None of
the proposed "biological measures" of pollution intensity could have
revealed the degree of such interference or damage. In order to obtain
the crucial evidence required, it would have been necessary to emphasize
the fisheries phase of the investigation.
It is not the purpose of this paper to discourage limnological research
pertinent to water pollution problems, nor is it intended to deny the value
of all biological indicators of pollution. There can be no doubt that a
drastic modification of any natural aquatic biota, attributable to a change
of water quality, can have highly undesirable aspects or consequences.
Such changes presumably are detrimental to human use and enjoyment of
natural waters more often than they are not. Many a readily demonstrable
effect of wastes upon aquatic life in a valuable stream is suggestive of
probable existing or incipient pollution which deserves close attention
and investigation. Even before valuable fish populations have been
materially affected by some potentially harmful pollutant, an observed
detrimental effect upon other organisms which are somewhat more
susceptible than fish may give warning of possible future damage to
Eisheries by continued or additional waste discharges. The nature and
ihe source of existing or incipient pollution also may be revealed by
appropriate biological indices. Finally, inasmuch as some of the organise
considered to be indicators of pollution are organisms which can directly
nterfere with human use or enjoyment of waters (e. g., unsightly slime-
arming organisms such as Sphaerotilus, odor-producing algae, etc. ),
heir unusual abundance may not be disregarded in evaluating over-all
lamage caused by pollution.
CONCLUSIONS
It must be concluded that every change or peculiarity of the flora
md fauna of a stream which has been referred to as an index or measure
>f pollution is in reality only an index of environmental disturbance or
invironmental anomaly. The disturbance or anomaly indicated may or
nay not be pollutional in the sense that stream uses are interfered with.
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Pollution (i. e. , interference with stream uses) can be negligible when
the effect on the aquatic biota as a whole is great, and it can be severe
when most of the aquatic life is unaffected. Gross pollution often can be
demonstrated without any biological investigation. When biological
investigation may be necessary, pollutional damage to valuable aquatic
organisms can probably best be determined by concentrating attention
upon these particular organisms. Yet, since all aquatic life forms are
more or less sensitive to changes of water quality, the fate of any of
them theoretically can be instructive, revealing something about the
nature and magnitude of these changes that may not be obvious nor
easily determined otherwise.
A genuine contribution to water pollution science can be made when-
ever the presence or relative abundance of living organisms of any kind
can be shown to be a reliable index of something tangible that one may need
to know in order fully to ascertain and understand the pollutional status
of an aquatic environment. When proposing and describing the use of
such biological indices, one should state specifically what it is that each
is believed to indicate, carefully avoiding such general, vague, or abstrad
terms as "pollution" and "stream health", which may be variously under-
stood. Does it indicate, for example, continual presence of dissolved
oxygen in certain concentrations believed to be adequate for sensitive
fish species? Does it indicate organic enrichment likely to interfere in
some way other than through oxygen depletion with certain specific uses
of water? Or does it indicate that particular toxic substances have not
recently been present in concentrations likely to be injurious to fish, to
man, or to certain crops? No simple biological indicator and no one
measure of stream conditions can indicate all of these things. But any
species can become a biological indicator of environmental conditions
of possible interest as soon as its nutritional and other environmental
requirements, its relative resistance to various toxic substances, etc.,
become known. Widely distributed sessile or sedentary organisms
should be the most useful indicators of past conditions. Unfortunately,
the water quality requirements of most of the "indicator organisms"
have never been thoroughly investigated, so that there is no real know-
ledge of specific factors which limit their distribution and abundance.
Probably nobody ntow knows just why any of the so-called clean-water
organisms begin to disappear from waters subject to progressively in-
creasing organic enrichment. Here is a field for future research which
is far more promising than is, for example, the questionable classificatio'
of all aquatic organisms as "pollutional", "clean-water", or "facultative"*
If there are common sedentary organisms whose water quality requireme*1
can be shown to correspond closely with those of valuable fish species, the
are potentially useful indicators. At the present time, however, exceptini
instances of gross pollution, only fish themselves can be said to indicate
reliably environmental conditions generally suitable or unsuitable for
their own existence.
L 59-
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REFERENCES
Beck, W.M.
1954. Studies in stream pollution biology. I. A simplified
ecological classification of organisms. Quarterly-
Journal of the Florida Academy of Science, 17:211-227.
Becks W. M.
1955. Suggested method for reporting biotic data. Sewage
and Industrial Wastes, 27:1193-1197.
Brinley, F. J. and L. I. Katzin
1944. Biological studies. Ohio River Pollution Control.
Supplements to Part 2, Report of the U. S. Public
Health Service. 78th Congress, 1st Session, House
Document No. 266, pp. 1275-1368.
California State Water Pollution Control Board.
1952. Water quality criteria. California State Water
Pollution Control Board, Publication No. 3.
Sacramento. (See also Addendum No. 1, 1954)
Cole, A.E.
1941. The effects of pollutional wastes on fish life.
In A Symposium on Hydrobiology, University of
Wisconsin Press, Madison, pp. 241-259.
Dimick, R. E. and F. Merryfield
1945. The fishes of the Willamette River system in relation
to pollution. Oregon State College Engineering
Experiment Station Bulletin Series, No. 20.
Doudoroff, P.
1951. Biological observations and toxicity bioassays in the
control of industrial waste disposal. Proceedings of
the Sixth Industrial Waste Conference, Purdue
University Engineering Extension Bulletin, Series No.
76, pp. 88-104.
Doudoroff, P.
In Press. Water quality requirements of fishes and effects
of toxic substances. In The Physiology of Fishes
(M. E. Brown, Editor)?" The Academic Press, New York.
Doudoroff, P. and M. Katz
1950. Critical review of literature on the toxicity of industrial
wastes and their components to fish. I. Alkalies,
acids, and inorganic gases. Sewage and Industrial
Wastes, 22:1432-1458.
-160-
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10. Doudoroff, P. and M. Katz
1 953. Critical review of literature on the toxicity of
industrial wastes and their components to fish. JI.
The metals, as salts. Sewage and Industrial Wastes,
25:802-839.
11. Ellis, M. M.
1937. Detection and measurement of stream pollution.
Bulletin of the Bureau of Fisheries, 48:365-437.
12. Fjerdingstad, E.
1950. The microflora of the river Molleaa with special
reference to the relation of the benthal algae to pollutior
Folia Limnologica Scandinavica No. 5.
13. Gaufin, A. R. and C. M. Tarzwell
1952. Aquatic invertebrates as indicators of stream pollution.
Public Health Reports, 67:57-64.
14. Gaufin, A. R. and C. M. Tarzwell
1953. Discussion of R. Patrick's paper on "Aquatic organisms
as an aid in solving waste disposal problems. " Sewage
and Industrial Wastes, 25i214-217.
15. Gaufin, A. R. and C. M. Tarzwell
1955. Environmental changes in a polluted stream during
winter. The American Midland Naturalist, 54:78-88.
16. Gaufin, A. R. and C, M. Tarzwell
1956. Aquatic macro-invertebrate communities as indicators
of organic pollution in Lytle Creek. Sewage and
Industrial Wastes, 28:906-924.
17. Harnisch, O.
1951. Hydrophysiologie der Tierp. Schweizerbart'sche,
Stuttgart. (Die Binnengewasser, Vol. 19)
18. Hasler, A. D.
1947. Eutrophication of lakes by domestic drainage.
Ecology, 28:383-395.
19. Johnson, J. W. H.
1914. A contribution to the biology of sewage disposal.
Journal of Economic Biology, 9:117-121.
20. Katz, M. and A. R. Gaufin
1953. The effects of sewage pollution on the fish population
of a Midwestern stream. Transactions of the American
Fisheries Society, 82:156-165.
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21. Katz, M. and W. C. Howard.
1955. The length and growth of 0-year class creek chubs
in relation to domestic pollution. Transactions of
the American Fisheries Society, 84: 228-238.
22. Kolkwitz, R.
1911. Biologie des Trinkwassers, Abwassers und der
Vorfluter. Rubner, Gruber und Ficker's Handbuch
des Hygiene. II. S. Hirzel, Leipzig.
23. Kolkwitz, R. and M. Marsson
1908. Oekologie der pflanzlichen Saprobien, Berichte der
Deutschen Botanischen Gesellschaft, 26a: 505-519.
24. Kolkwitz, R. and M. Marsson
1909. Oekologie der tierischen Saprobien. Internationale
Revue der gesamten Hydrobiologie und Hydrographie,
2: 126-152.
25. Needham, P. R. and R. L. Usinger
1956. Variability in the macrofauna of a single riffle in
Prosser Creek, California, as indicated by the
Surber sampler. Hilgardia, 24: 383.-409.
£6. Patrick, R.
1949. A proposed biological measure of stream conditions,
based on a survey of the Conestoga Basin, Lancaster
County, Pennsylvania. Proceedings of the Academy of
Natural Sciences of Philadelphia, 101: 277-341.
17. Patrick, R.
1950. Biological measure of stream conditions. Sewage and
Industrial Wastes, 22: 926-938.
18. Patrick, R.
1953. Biological phases of stream pollution. Proceedings
of the Pennsylvania Academy of Science, 27: 33-36.
19. Patrick, R.
1953. Aquatic organisms-as an aid in solving waste disposal
problems. Sewage and Industrial Wastes, 25: 210-214.
10. Patrick, R., M. H. Hohn, and J. H. Wallace
1954. A new method for determining the pattern of the diatom
flora. Notulae Naturae, No. 259.
1. Richardson, R. E.
1928. The bottom fauna of the Middle Illinois River, 1913-1925
Illinois Natural History Survey Bulletin, 17: 387-475.
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32. Ricker, W. E.
1934. An ecological classification of certain Ontario
streams. University of Toronto Studies, Biological
Series No. 37.
33. Southgate, B, A
1948. Treatment and disposal of industrial waste water.
Department of Scientific and Industrial Research
(Gr. Brit. ), London.
34. Steinmann, P. n
1915. Praktikum der Susswasserbiologie. Verlag Borntraeger,
Berlin.
35. Suter, R. and E. Moore
1922. Stream pollution studies. Bulletin, New York State
Conservation Commission.
36. Tarzwell, C. M. and A. R. Gaufin
1953. Some important biological effects of pollution often
disregarded in stream surveys. Proceedings of the
Eighth Industrial Waste Conference. Purdue University
Engineering Extension Bulletin, Series No. 83, p. 295-3
37. Thienemann, A. ,,
1925. Die Binnengewasser Mittqjeuropas. Schweizerbart1 sche
Stuttgart. (Die Binnengewasser, Vol. 1)
38. Wurtz, C. B.
1955. Stream biota and stream pollution. Sewage and
Industrial Wastes, 27: 1270-1278.
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biological criteria for the determination of lake pollution
Eugene W. Surber *
Michigan Water Resources Commission
Lansing, Michigan
Many, if not most of our productive warm-water lakes have low-
oxygen conditions existing in them through at least a part of the summer.
At this time, conditions prevail that closely approach conditions caused
artificially by domestic or industrial pollution. In the eutrophic "chironomus"
lakes of Connecticut described by Deevy (1941), one part per million of
°xygen or less prevailed from late June until early November, but zero
°xygen was not recorded.
The fact that aquatic earthworms of the family Tubificidae (Limno-
jjilus and Tubifex in this report) and such red midgefly larvae as Tendipes
^gcorus, Tendipes tentans, Tendipes plumosus and Tendipes riparius are
able to live in water of very low oxygen content is well known. Lindeman
U941) demonstrated that "chironomus" larvae could withstand complete
l^ck of oxygen for four months at 10°C, but Deevy (1941) pointed out that
^uch of the case for existence of these organisms under anerobic con-
ditions rests on analyses made with either the unmodified Winkler method
0r the Rideal-Stewart modification. Recently, Ruttner (1953) cautioned
t}lat Alsterberg's modification of the Winkler method should be used for
°xygen determinations otherwise values are lower due to the presence of
^educing substances.
This report stemmed from a study of American papers dealing with
the quantitative abundance of aquatic earthworms in both lakes and streams.
!t was concluded last year in a report to the Midwest Benthological Society
*kat in large unpolluted lakes, such as Lake Michigan, Lake Nipigon,
Douglas Lake (Michigan), etc. , the average number of tubificids per
8
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Hexagenia (a burrowing mayfly) per square foot, lightly polluted 9.4-93
tubificids per square foot; moderately polluted 94-456 tubificids per square
foot; and heavily polluted over 456 tubificids per square foot.
In regular survey work, the writer has been in the habit of looking cri-
tically at each sample, identifying the animals present to species if possible,
and classifying them as pollution-tolerant, facultative, or clean-water. Each
sample, even in a lake where there are usually few species, is capable of
revealing conditions in a limited area, especially where the information is
correlated with data on oxygen content of the water, biochemical oxygen demai*
(B.O.D-), other chemical analyses, temperature, etc., collected at the same
time.
With the exception of a few samples from White Lake and Saginaw Bay,
the bottom samples were collected with a 6 x 6 inch Ekman dredge, and the
bottom materials were sieved through a No. 30 sieve.
Main reliance has been placed on bottom samples collected from the
deeper waters of lakes outside the littoral zone of vegetation, for it seemed
obvious that damage to a lake by pollution would show up there first. While
the shoreward zones of vegetation contain a greater variety of organisms,
the photo synthetic activity of plants in polluted areas and the circulation of
surface waters are likely to create better living conditions in the zone of
vegetation than exist in waters deeper than about 15 feet. In the lakes sampled
thus far, the deeper-waters comprised most of the lake area. It is believed
that conditions must be maintained in a satisfactory condition there if these
lakes are to remain productive of fish and fish food organisms.
Since White Lake in Muskegon County, Michigan, had from 8 to 17
species of bottom animals at depths to 60 feet; Muskegon Lake 6 to 8 species
at depths to 61 feet; Lake Charlevoix in its less-polluted areas near Boyne
City from 3 to 9 species down to depths of 44 feet, it was decided to take
a collective look at the lake bottom sampling in the Michigan lake pollution
surveys to determine (1) whether more than 100 tubificids per square foot
actually represented polluted conditions, (2) what other organisms of the
deeper waters might be present and classed as pollution-tolerant or hardy,
sensitive or intermediate, (3) any other useful criteria of lake pollution
such as comparisons of total numbers of animals, exclusive of the tubificids.
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This "collective look" brought regrets that more samples were not taken
in many places, but the presentation of the data collected may be of interest
and some value in establishing norms and methods of sampling in the future
which may lead to quantitative numerical values or norms above or below
"which damage to bottom animal life is more apparent.
Manistee Lake may be cited as an example of a lake in which even tubi-
ficid worms could not live in the deeper waters. In this lake, 17 of 24 samples
collected September 22, 1954, were entirely without bottom animals of any
kind. This lake has been severely polluted with paper mill and other wastes .
The deeper areas of Fremont Lake, likewise, were apparently severely
damaged by organic pollution exceeding the quantity the lake could assimilate,
md many of the samples were without tubificid worms and other species
normally present. On September 16, 1952, 4 of 14 samples were entirely
vithout animals . A neighboring unpolluted lake, Pickerel Lake, sampled
September 15, 1952, of similar depth and without oxygen in its deepest waters
iad three times as many bottom animals. There were about three times
ls many Tendipes plumosus and ten times and many Chaoborus in the
>ottom samples .
In Saginaw Bay, the key area of Lake Huron from a fisheries standpoint,
amples were taken at widely scattered points, but it has now developed that
hese were not collected at the best season; collection apparently followed
tie emergence of Michigan "caddis" Hexagenia limbata and the "bloodworm"
'endipes plumosus, the young of which occurred in several samples.
Saginaw Bay receives pollution from the Saginaw River which carries
ldustrial and municipal wastes from Midland, Saginaw and Bay City. Minor
ontributions of pollution are made to the bay by the Kawkawlin and Pinconning
ivers. A beet sugar plant at Sebewaing introduces wastes seasonally into
te bay. There is also a milk plant located at Sebewaing which discharges
eated wastes into Sebewaing Bay. The City of Saginaw began operating
new sewage treatment plant before the 1954 survey. Pollutants peculiar
the operation of a large chemical plant at Midland include brine and phenolic
astes . The latter have been reduced considerably in recent years . Chlorides
Lnged from about 25-50 parts per million in the bay.
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Data on Green Bay, the key area of Lake Michigan, have been included
for comparison with Saginaw Bay, although sampling there occurred at a more
favorable time, prior to June 1 . The area of Green Bay referred to as the
"Lower Bay" in Table I is closest to the polluted Fox River which has many
paper mills located on its banks between Green Bay and Appleton, Wisconsin.
The series of samples in the Oconto line probably represent relatively
unpolluted conditions in spite of the fact that the lower Oconto River is heavily
polluted by paper mill wastes .
White Lake, at Whitehall and Montague, Michigan, may be classed as
virtually unpolluted. The surveys made of it both preceded and followed the
construction of a large chemical plant which provided adequate waste treatment
facilities .
Ellsworth Lake, Antrim County, Michigan, a lake in a chain of connected
lakes receives canning wastes seasonally. When it was sampled on October
8, 1953, most of the lake had 0.2 part per million or less of dissolved oxygen.
Because of the shortage of oxygen, crayfish, which were abundant in the lake
at the time, had migrated from the deeper water to the very margin and some
even out of the water in an effort to survive low oxygen conditions . Many of
them died at the water's edge. Freshly dead midgefly larvae of Tanytarsus
nigricans were found in only two bottom stations, but many had risen to the
water surface where they were fed upon by gulls. Larvae of the biting midge
Palpotnyia tibialis * were numerous in these bottom samples in spite of the
low oxygen, and apparently many of them left the bottom temporarily for
only 55 percent of the samples contained them on October 8, 1953. When the
lake was revisited February 10, 1954, they were twice as abundant as during
the sampling of October 8, 1953, and 91 percent of the samples contained
them.
Tendipes plumosus larvae occurred in 64 percent of the Ellsworth Lake
samples on October 8, and in 55 percent of the samples collected from under
ice cover February 10, 1954, indicating that most of them were hardy enough
to survive the temporary period of low oxygen in the lake.
A series of three stations across the lower end of St. Clair Lake located
in the Intermediate River chain of lakes immediately above Ellsworth Lake
porvided a small amount of data from an unpolluted area to compare with
* identified for the writer by Dr. Willis Wirth, U. S. National Museum.
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data from Ellsworth Lake. One of the principal differences was low average
number of tubificids, average 9.3 per square foot in St. Clair Lake compared
to 52.4 in Ellsworth Lake on October 8, 1953, and an average of 8 tubificids
in St. Clair Lake compared with 63 per square foot in Ellsworth Lake on
February 10, 1954. Larvae of Calopsectra dives were present in St.Clair
Lake samples and absent from Ellsworth Lake.
Paper mill wastes enter Muskegon Lake on the south shore, and in that
vicinity tubificid worms were particularly numerous. They were nearly
ten times as numerous as they were in the control area on the north side of
the lake. Midgefly larvae of Procladius and Tendipes plumosus were present
in small numbers in at least half of the samples on both sides of the lake.
Lake Charlevoix at Boyne City is polluted by wastes from a large tannery
and by the untreated wastes of the city. An extensive area of the lake bottom
near Boyne City contains hair (originating in the tannery wastes). The bottom
sampled was roughly divided into two areas. The area of maximum pollution
adjacent to the tannery and city had deposits of hair ranging from 2,000-6,950
bounds per acre on a dry weight basis. The area of minimum pollution coincided
vith hair deposits of less than 2,000 pounds per acre. Tubificids were most
abundant in the zone of greatest hair deposits. Midges of Procladius were also
nost abundant in this zone.
Hart Lake is an artificial impoundment on the South Branch of the Pent-
vater River. It receives cannery wastes seasonally, and the samples collected
here were taken at a time (July 22, 195 3) when cherry canning had been in
rogress for some time. It receives untreated domestic sewage from a city
f about 2,200 population. Probably the point of greatest interest in this
tudy was the abundance of Chaoborus punctipennis in the samples collected
elow the sources of pollution. They ranged in number from 16-504 per square
>ot, with three samples containing 296, 324, and 504 per square foot. Larvae
f Tendipes tentans-plumosus and Procladius were numerous in some
amples.
There are many details of description and notes on abundance of species
iat might have been noted in this report; however, the relatively small
amber of samples taken in most cases does not justify the drawing of many
inclusions at present.
.Ua.
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The following quantitative data are presented primarily to show trends
of abundance of species in polluted areas. Tables 1 and 2 show average numbef
of species in each area; total number of animals in all samples of each area;
average number of animals per square foot, both with and without tubificid
worms; average number of tubificids per square foot, abundance of several
species of midgefly larvae, phantom midges, and fingernail clams .
Tables 3 and 4 show the frequency of occurrence of the above kinds of
bottom animals in all samples in percentages of the number of samples taken.
CONCLUSIONS
A survey of the lake reports showed that an abundance of tubificids in
excess of 100 per square foot apparently truly represented polluted habitats.
Severe pollutions such as occurred in a depression in Fremont and in vir-
tually all of Manistee Lake precludes all forms of animal life. Such data
must be approached with care when statistical analyses are considered because
considerable pollution may cause increased abundance in some areas and
severe pollution may eliminate or reduce numbers of even the hardy forms
in other areas.
The data show that Procladius species are usually present with tubificids
in polluted areas, but in severe pollution only the tubificids are able to survive-
Palpomyia tibialis was found to be a very hardy species in one lake where
conditions brough about the destruction of Tanytarsus nigricans.
The fingernail clam Pisidium sp. was more prevalent in polluted areas
than Sphaerium or Musculium, and judging from the abundance of dead shells
in grossly polluted areas, these small mollusks are not able to survive the
adverse conditions tolerated by tubificids .
Gryptochironomus digitatus and Calopsectra dives are common associates
of Tendipes plumosus or Tendipes tentans, tubificids and Procladius in the
deeper waters of the polluted areas sampled. It is presumed that they are
hardy, although their hardiness is apparently less than either tubificids or
Procladius. The greater average abundance of Procladius in polluted habitats
is presumed to be due in part to the fact that they feed upon tubificids.
Chaoborus punctipennis may frequent rather heavily-polluted areas and
become abundant there. Although their swimming ability and habit of migrating
to the surface at night permits them to leave polluted areas in which tubificids
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either die or thrive, there is some evidence, as in the comparison of Pickerel
Lake (an unpolluted lake) and Fremont Lake (a polluted lake) and the north
and south sides of Muskegon Lake, that Chaoborus can not survive in large
numbers in a polluted lake where low oxygen conditions have apparently
prevailed for an excessive period of time.
When average numbers of bottom animals per square foot including tubi—
ficids are compared with average numbers less tubificids, most populations
reveal the large contribution of the latter numerically. The difference may
also reflect the contribution of organic wastes as a fertilizer in lakes as
it did in lower Hart Lake.
Number of species in lakes as well as streams appears to be the most
reliable criterion of pollution. Unpolluted Michigan lakes from these limited
observations appear capable of supporting a number of species even to depths
of 60 feet.
Deevy, E. S., Jx.
1941 Limnological studies in Connecticut, VI. The quantity and composi-
tion of the bottom fauna . Ecological Monographs, Vol. II, pp . 413-455 .
Lindeman, Raymond L. ,
1941 Seasonal food dynamics in a senescent lake. American Midland
Naturalist, Vol. 26, No. 3, pp. 636-673.
Ruttner, Franz (translated by D. G. Frey and F. E. J. Fry)
1953. Fundamentals of Limnology. University of Toronto Press, 242 pps .
Original German Edition published by Walter de Gruyter & Company, Berlin,
1952.
Wright, Stillman and Tidd, Wilbur M.
1933. Summary of limnological investigations in western Lake Erie in
1929 and 1930 . Trans . Amer . Fish. Society, Vol. 63, pp. 271-285 .
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TABLE I. AVERAGE NUMBERS OF BOTTOM ANIMALS PER SQUARE FOOT IN POLLUTED MICHIGAN LAKES
A.
Saginaw
Saginaw
Green Bay
Green Bay
White Lake
White
Ellsworth St. Clair
Ellsworth
St. Clair
Bay July
Bay July
(Wis.)
Oconto line
May26,27
Lake
Lake Oct
. Lake Oct.
Lake Feb.
Lake Fel
25-26
16-17
Lower Bay
May 26,
1954
Aug J 9
8, 1953
8, 1953
10, 1954
10, 1954
1953
1954
May 27,
1952
1952
1952
Number of Samples
8
14
20
7
22
16
11
3
11
3
Average number of species
5.8
9.1
9.7
6.3
12
7.4
6.1
15
5.6
12.7
Total number of animals (all
samples)
1,908
3,339
25,332
1,212
10,504
4,163
1,672
824
1,868
629
Average number per square foot
239
239
1,267
173
478
260
152
275
170
210
Aquatic earthworms
Average number of tubificid worms 162
130
1,084
35
98
60
52
9
63
8
Average number less tubificids
77
109
183
138
380
200
100
266
107
202
Midges
Av .number Procladius sp.
7.8
5.1
65 2
44.0
53.8
46.0
23.3
44
36
60 »
Av .number Cryptochironomus
N
digitatua
6.0
2.8
7 2
2.9
5.7
5.3
0.4
0.0
0.0
4.0 T
" " Tendipes tentans-plumosus 6.0
22.3
16.0
14.8
46.8
54 .3
5.1
0.0
7.6
2.7
" " Calopsectra dives
6.3
9.0
162
32.6
26.1
12
0.0
12.0
0.0
20.0
" " Felopia stellatus
0.0
0.0
6.3
0.0
0.0
0.0
0.0
0.0
0.0
0.0
Phantom midges
Av . number Chaoborus punctipennis 0.0
0.0
0.0
0.0
602
10.8
24.4
10.7
19.3
6.7
Punkies
Av. number Palpomyia sp.
0.0
0.1
0.0
0.6
4.8
0.0
13.5
a.o
32.0
4.0
Fingernail clams
Av. number Pisidiuxn
2.0
3.1
41.3
26.8
14.1
8.9
1.5
2.7
0.7
4.0
" " Sphaerium-Musculium
0.0
1.1
8.3
4.0
1.9
0.8
0.7
0.0
0.7
0.0
-------
TABLE 2. AVERAGE NUMBERS OF BOTTOM ANIMALS PER SQUARE FOOT IN POLLUTED MICHIGAN LAKES
Muskegon Muskegon L.Charle- L.Charle- Pickerel Fremont Manistee Manistee Hart Lake Hart Lai
Lake. Lake Pol- voix.Zone voix.Zone Lake Lake Lake Lake above below
Northside luted Area o£ least of max. Sept. 15 Sept. 16 Sept. 15 Sept. 22 pollution pollution
Sept. 15, Sept. 15, pollution pollution 1952 1952 1953 1954 July 22, July 22,
1954 1954 May 19, May 19, 1953 1953
1954 1954
Number of samples
4
12
11
12
8
14
24
24
2
8
Average number of species
8.2
5.8
5.7
7.3
3.6
1.5
0.95
0.8
11.5
4.8
Total number of animals (all
samples)
2,396
26,659
1,060
5,839
368
228
720
556
504
5,314
Average numbers per square
foot
599
2,222
96
487
46
16
30
23
252
664
Aquatic earthworms
Average number of tubificid
worms
227
2,092
19
344
1.0
5.0
18
16
76
373
Average number less tubificids
372
130
77
143
45.0
11 .0
12
7
176
291
Midges
Av . number Procladius sp.
9-0
10.7
39.3
60.3
2.5
2.0
0.2
1.2
6.0
37,5
'! " Cryptochironomus
digitatus
0.0
2.3
2.5
1.7
0.0
0.1
0.2
0.2
1.7
12.0
" " Tendipes tentans-plumosus 9.0
8,3
0.7
6.0
9.5
3.4
0..0
0.0
10.0
71.7
" 11 Galopsectra dives
1.0
0.0
3.3
0.7
0.0
0.0
0.2
0.0
2.0
0.0
" " Pelopia stellatus
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
1.5
Phantom midges
Av . number Chaoborus puncti-
penms
46.0
2.3
0.0
0.0
16.0
0.6
0.0
0.0
2.0
163
Punkies
Av . number Palpomyia sp.
0.0
0.0
0.0
0.0
0.5
0.0
0.0
0.0
4.0
0.0
Fingernail clams
Av. number Pisidium
15.0
14.7
6.9
12.6
1.0
0.0
0.5
0.8
40.0
3.5
" " Sphaerium-Musculium
7.0
9.0
0.4
10.0
0.0
0.0
02
0.0
22 .0
12.5
-------
TABLE 3. FREQUENCY OF OCCURRENCE OF VARIOUS SPECIES IN LAKE SAMPLES EXPRESSED AS THE PERCENTAGE OF THE TOTAL
SAMPLES TAKEN
Saginaw Saginaw Lower Green Green Bay White L. White L. Ellsworth St. Clair Ellsworth St. Clair
Bay-July Bay-July Bay, May 27 Oconto line May Aug. 19, Lake Oct. Lake Oct. Lake Feb. Lake Feb.
25, 26 16.17 1952 May 26, 1954 1952 8, 1953 8,1953 10, 1954 10, 1954
1953 1954 1952
Tubific idae
100
100
100
71
95
88
82
67
91
67
Procladius
63
71
95
100
100
94
73
100
82
67
Cryptochironomus digitatus
63
71
80
80
64
50
9.1
0
0
67
Tendipes tentans-plumosus
13
79
70
43
55
69
64
0
55
33
Chaoborus punctipennis
0
0
0
0
64
56
91
33
64
33
Calopsectra dives
50
43
45
86
86
19
0
100
0
67
Pelopia stellatus
0
0
15
0
0
0
0
0
0
0
Palpomyia
0
14
0
14
36
0
55
67
91
6?
Pisidium
25
50
65
86
86
44
18
33
9
33
Sphaerium and Musculium
0
21
40
43
23
6
9.1
0
18
0
-------
TABLE 4. FREQUENCY OF
OCCURRENCE OF VARIOUS SPECIES IN LAKE SAMPLES EXPRESSED AS THE PERCENTAGE OF THE TOTAL
SAMPLES TAKEN
Tubific idae
Procladius
Cryptochironomus digitatus
Tendipes tentans -plumosus
Chaoborus punctipennis
Calopsectra dives
Pelopia stellatus
Palpomyia
Pisidium
Sphaerium and Musculium
Muskegon Muskegon
Lake Lake
North polluted
Side area
Sept. 15, Sept. 15,
1954 1954
Lake Charle- L. Charle-
voix. Zone
of least
pollution
May 19,
1954
voix. Zone
of great-
est pollu-
tion May
19, 1954
Pickerel
Lake
Sept. 15,
1952
Fremont
Lake
Sept. 16,
19S2
Manistee
L. Sept.
15, 1953
Manistee
Lake
Sept. 22,
1954
Hart Lake
above pol-
lution
source
July 22,
1953
Hart Lake
below pol
lution
sources
July 22,
1953
100
92
91
100
13
29
17
21
100
100
75
75
100
100
25
21
4
8
100
88
0
8.3
45
33
0
7
4
4
100
25
75
50
18
42
75
36
0
0
100
72
50
25
0
0
75
7
0
0
50
100
25
0
27
17
0
0
4
0
50
0
0
0
0
0
0
0
0
0
0
25
0
0
0
0
13
0
0
0
50
0
75
25
18
42
25
0
4
4
100
25
50
33
45
17
0
0
4
0
100
50
-------
THE USE AND ABUSE OF INDICATOR ORGANISMS
William M. Beck, Jr.
Florida State Board of Health
Jacksonville, Florida
In the foregoing discussions, several methods for the use of indicator
organisms for many taxonomic groups have been presented. The discussions
have been perhaps more enlightening than the formal papers. As last speaker
I have the advantage of having heard all the previous papers and discussions.
From all we have heard, it would appear that several points remain to be
discussed.
Throughout the foregoing presentations, the term "biological indicators
pollution" has been widely - albeit loosely - used. Most of the speakers
igreed that individual species as indicators of pollution do not exist, yet
;he entire day was devoted to indicator organisms.
The indicator organism program which we use in Florida is based on
Organisms which are indicative of the absence of organic pollution rather than
its presence. Such a program has served our needs very well. Separate
lPproaches or classifications are needed in the case of chemical or physical
^llutants. These are developed as the need arises.
There appear to be many opinions as to just what an indicator organism
'rogram is for. Our indicator organism program in Florida is but a part
'f the overall stream sanitation program. The amount we can learn from the
'istributions of the stream inhabitants is great. From some of the questions
'hich have been asked here today, it appears obvioussthat at least a few
eople came here seeking a simple indicator organism program which would
olve all their problems. It should be obvious that no such classification of
rganisms exists.
The question naturally arises, then, as to just what is to be expected
¦oiti an indicator organism program. Such a program may prove an excellent
3ol, with the limitations of a tool in that it may, when handled properly,
0 some jobs well while proving useless for others.
Perhaps a review of the use of our program in Florida will clarify the
sneralities stated above. As has been pointed out in literature (Beck, 1954)
more widely distributed stream organisms have been classified with re-
*r"d to organic pollution in what, historically at least, would appear to be
backward manner. Instead of indicating pollution, our methods are set
3 "with regard to absence of pollution. The former concept was found to be
tenable - as most of the previous speakers have indicated.
In general stream surveying the indicator organism method is used in
^junction with the normal chemical, physical and bacteriological methods.
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The biological program is thus supplemental to the more widely established
methods. From it we obtain the evidence of yet one more discipline. The
fauna of a stream is the result of the combined chemical, biochemical, phy-
sical, biotic, climatic and geologic factors of the area in which the stream
occurs. Chemical or physical damage to a stream may be a rather
ephemeral thing, but biotic evidence of such damage may persist for a signi-
ficant period. Herein lies one of the main values of an indicator organism
program, that of, if I may risk approaching the ridiculous, "faunal memory".
The use of indicator organisms in making a quick check of the condition
of a stream is of particular value. In the Biotic Index paper {Beck, 1955:
1196) figure 3 was included to show that the results of surveys made one
year apart were quite similar when conditions remained constant. Results
since then are most interesting. All data are for station 2 which is located
about two miles below the source of waste.
Year Biotic Index
1953 5
1954 2
1955 16
1956 31
It wquld appear that station 2 was in poor condition in 1953 and in
slightly worse condition in 1954. However, a quick check of the stream in
1955 showed evidence of a significant biotic change indicative of less pollu-
tional effect. A visit to the industrial plant revealed that major changes
had been made which had already helped the river. Actual work on the
stream which revealed this change took less than an hour. The 1956 result
reveals that the stream has completely recovered.
The disappearance of a snail (Goniobasis sp.) was the first evidence
we had of a new source of pollution in another river. No single classifica-
tion of organisms will suffice for all aspects of stream pollution. The
pollution of two rivers with wastes high in fluoride ion content necessitated
a new classification of indicator organisms. Some species which were
quite'tolerant of organic pollution proved incapable of surviving even moder-
ate concentrations of the fluoride ion. One of the most fluoride-resistant
insects is normally a clean water species.
I believe that it has been stated here that indicator organisms seemed
to be indicative of dissolved oxygen concentration. We found it impossible
to classify stream invertebrates in Florida with regard to dissolved oxygen
content of the water. The presence of a great many springs along such rive**
as the Suwannee and the St. Johns greatly alters the oxygen picture without
damage to the fauna. As a result we find the more sensitive species of
invertebrates tolerating surprisingly low oxygen tensions.
-176-
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It should be stated that the methods we use in Florida are, we feel,
useful. They are subject to revision any time better methods are suggested.
They are essentially beginning methods and are being studied, subjected to
testing and revised almost constantly.
In conclusion it should be stated that nothing has been written or spoken
which has in any way shaken my belief in the usefulness of a properly
developed indicator organism program as long as the user realizes that
any system, no matter how carefully developed, has its limitations. No
such system will ever be a substitute for a knowledge of the behavior of
streams and of the inhabitants of streams .
References Cited
Beck, William M., Jr. 1954. Studies in stream pollution biology. I. A
simplified ecological classification of organisms. Quart. Journ. Fla . Acad.
Sci., 17(4):2 11-227 .
1955. Suggested method for reporting biotic data.
Sewage and Ind. Wastes , 27(10): 1193-1197 •
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CURRENT INVESTIGATIONS IN WATER POLLUTION BIOLOGY
-------
Current Water Pollution Investigations and Problems
in Wisconsin
by
Kenneth M. Mackenthun
Wisconsin Committee on Water Pollution
Madison, Wisconsin
The Wisconsin statues define the term "pollution" as the "contamination
or rendering unclean or impure the waters of the state, or making the same
injurious to public health, harmful for commercial or recreational use, or
deleterious to fish, bird, animal, or plant life".
The major problems in stream pollution concern themselves principally
¦with domestic wastes and industrial wastes. At the present time, about
94. 8% of the population served by sewers is connected to treatment plants.
In 1949, 62 communities still discharged untreated sewage. As of January 1,
*956, this number has been reduced to 29, and of these, 5 have plants under
construction. There are a total of 319 treatment plants serving 406 com-
munities.
The 35 pulp and paper making firms in Wisconsin operate 45 paper
mill8, 16 sulphite pulp mills, 5 kraft pulp mills, and 14 groutidwood mills.
There are 1, 766 dairy plants which include some 1,064 cheese factories.
In addition, there are 151 canning plants as well as a full complement of
other industries. The total sources of pollution in Wisconsin at the pre-
sent time, number 1, 086.
The history of water pollution control in Wisconsin dates back to
about 1925. Prior to 1925, there was a limited amount of pollution control
through the application of statutes granting certain powers and duties to
the State Board of Health, and conservation statutes prohibiting the dis-
charge of certain specified materials to streams.
In 1925, the legislature appropriated $10, 000 from conservation funds
for water pollution control. This was expended in cooperation with the
State Board of Health in a study of major problems.
The legislature in 1927 created the Committee on Water Pollution,
Consisting of the State Chief Engineer, a member or representative of
he Public Service Commission designated by the Commission, a Con-
servation Commissioner or an employee designated by the Conservation
-179-
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Commission, the State Health Officer or a member of the Board of Health,
and the State Sanitary Engineer or other engineer appointed by the Board
of Health. The State Board of Health was designated as the administration
agency of the Committee.
The revision of the 1927 act by the 1949 legislature recognized the
need for a full-time water pollution control program and provided for a
full-time director and authorized an appropriation which made possible
the employment of a full-time staff. The present staff consists of the
Director, an industrial wastes engineer, an engineer in charge of coordi-
nating stream surveys, a biologist, a chemist, and four field engineers.
The Committee's basic responsibility is to exercise general super-
vision over the administration and enforcement of all laws relating to the
pollution of the surface waters of the state, and to study and investigate
all problems connected with the pollution of surface waters of the state
and its control, and to make reports and recommendations thereon. It
further has the authority:
"To conduct scientific experiments, investigations, and research to
discover economical and practicable methods for the elimination,
disposal, or treatment of industrial wastes to control pollution of
surface waters of the state. To this end, the Committee may co-
operate with any public or private agency when requested by such
an agency in the conduct of such experiments, investigations, and
research, and may receive on behalf of the state any moneys which
any such agency may contribute as its share of the cost under such
cooperative arrangements.
"To supervise chemical treatment of waters for the suppression of
algae, aquatic weeds, swimmers' itch, and other nuisance-producing
plants and organisms.
"To issue general orders and adopt rules and regulations applicable
throughout the state for the installation, use, and operation of practi-
cable and available systems, methods, and means for controlling the
pollution of the surface waters of the state through industrial wastes,
refuse, and other wastes.
"To issue special orders directing particular owners to secure such
operating results toward the control of pollution of the surface waters
as the Committee may prescribe within a specified time.
"To make investigations and inspections to insure compliance with
any general or special orders, rules and regulations which it may
issue.
-180-
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"To enter into agreements with the responsible authorities of other
states subject to approval by the Governor relative to methods, means,
and measures to be employed to control pollution of any inter-state
streams and other waters, and to carry out such agreements by
appropriate special and general orders.
"In addition to all other powers and duties of the Committee on Water
Pollution, it shall have the power and it shall be its duty to hold a
public hearing relative to alleged water pollution upon the verified
complaint of six or more citizens filed with the Committee"
During the course of development, cooperative programs of investi-
gation were established with the Sulphite Pulp Manufacturers' Research
League, the Pulp and Paper Advisory Committee on Waste Disposal, the
National and Wisconsin Canners' Associations, and recently cooperative
biological surveys have been conducted with the Institute of Paper Chemistry.
The Sulphite Pulp Manufacturers' Research League has been con-
tinuously engaged in research on waste disposal problems of the industry
since its formation in 1939. Spent sulphite liquor is produced when wood
is cooked for production of the pulp which is used in the manufacture of
paper. For each ton of pulp produced, about one ton of solids is contained
in the spent liquor which was formerly discharged directly to the streams.
One of the first methods developed for the utilization of waste sulphite
liquor was that of aerobic fermentation leading to yeast production. Today
there are two full-scale yeast plants, one at Rhinelander, Wisconsin pro-
ducing 14, 000 pounds of yeast per day, and one at Green Bay, Wisconsin
designed to produce 28, 000 pounds of yeast per day. Other processes
upon which much work has been done include evaporation and burning of
which there are three installations in Wisconsin, the production of vanillin,
the use of waste sulphite liquor in roadbinding of which about 60 million
gallons is used per year, soil filtration, and others of less potential.
In 1949 when the water pollution control statutes were revised, it was
recognized that some of our streams were still being adversely affected
by raw sewage and certain industrial wastes. At that time, the state was
divided into work areas comprising the 28 major drainage basins, and %
field engineer was assigned to each basin and instructed to conduct surr
veys to determine the condition of the streams and the sources of pollution
discharging to the streams. An industrial waste census survey and the
data on stream conditions above and below sources of pollution compiled
into a comprehensive report, including biological data taken abpve and
-¦I 81 -
-------
below the various sources, then became available for each basin studied.
This report, used as an exhibit at a public hearing, provided information of
value to the Committee in deciding on the types of orders to be issued in
order to eliminate or reduce pollution. The Committee has over a period
of years issued 1, 086 orders of which 316 have been fully satisfied as of
January 1, 1956.
The role of a biologist in the program follows two principal lines or
activity. The Committee is charged with the supervision of all chemicals
placed in the public waters of the state for the control of algae, weeds,
swimmers' itch, or other nuisance-producing organisms. The adminis-
tration and supervision of the aquatic nuisance control program thus be-
comes the responsibility of the biologist. During the summer of 1955,
for example, some 37, 000 pounds of copper sulphate was used for algae
control in 15 state lakes. In addition, on an experimental basis, 60 gallons
of Cutrine, 90 gallons of Delrad, and 400 pounds of Phygon were used for
algae control. The submergent aquatic vegetation control program en-
tailed the use of some 18, 800 gallons of commercial sodium arsenite
solution in 34 state lakes. In addition, a small quantity of 2, 4-D,
Oalapon, and methoxone chlorax was used. The control of snails har-
boring the organism producing swimmers' itch on some of the bathing
areas of the state involved the use of 1740 pounds of copper sulphate,
550 pounds of copper carbonate, and 320 pounds of lime.
The other phase of principal biological activity lies in stream sur-
vey work. In the past, the stream study program has consisted of
biological samples, principally those of bottom organisms, bracketing
the sources of pollution in the 28 drainage basins. From 1949 to the
present time, some 37 such surveys have been completed. The entire
state has now been covered and the initial phase of the pollution investi-
gation program completed.
The current and future biological program is aimed at more in-
tensive studies in more localized areas on the major waterways of the
state. Biologists of the Institute of Paper Chemistry at Appleton,
Wisconsin have also initiated a program of intensive biological study on
some of the rivers affected by the pulp and paper industry.
A recent cooperative biological study with the Institute of Paper
Chemistry has been completed on the lower Fox River in the central
portion of the state. The river is 39 miles long, has a fall of 4. 25
-182-
-------
feet per mile, and contains 14 dams. Effluent from 19 paper making pro-
cesses enters the river at various points and includes the effluent from
5 de-inked mills, 2 rag pulp mills, 1 kraft mill, 4 sulphite mills, and 2
groundwood mills. In addition, there is the effect from algae as tests
have indicated an excess of 130 tons per day (dry weight) of algae flowing
into the Fox River from Lake Winnebago.
Another biological study of the more intensive type was recently
completed on a 200-mile stretch of the Wisconsin River. This section
receives effluent from pulping processes of 5 sulphite mills, 3 kraft
mills, 2 semi-chemical pulp mills, 1 rag pulp mill, and 3 groundwood
mills. The combined pulp production is some 1480 tons of pulp per day,
and the combined daily paper production is some 2300 tons. A follow-up
partial survey in the late winter of some of the clean-water, summer-
time stations indicated quite drastic changes and severe conditions. It
is apparent that nearly as much attention must be given to the study of a
stream in winter as well as under summer-time conditions.
The Botany Department of the> University of Wisconsin has been
continuing its efforts in the field of nutritional requirements of blue-
green algae. At the same time, a continuous study on the removal of
nutrients from sewage effluent by the use of algae is conducted. The
study involves nutrient balances, biological examinations, and methods
of algal harvesting. Three ponds with a combined area of one-half acre,
and with controlled physical factors (flow, depth, and recirculation), are
being used in this investigation.
183-
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INVESTIGATIONS AND PROBLEMS IN ONTARIO
John H. Neil
Ontario Department of Health
Toronto, Canada
In Canada, relatively few biologists are working exclusively in the field
of pollution control and water supply. Consequently, it is necessary that
the studies include most phases of pollution biology as is evident from the
folbwii^g discussion of "Investigations and problems in Ontario."
Pollution control in Ontario is generally directed by the Pollution Contri
Board, which is composed of members from various interested govern-
ment departments . The actual investigations and control are the responsi-
bility of the Sanitary Engineering Division of the Department of Health and
the Fish and Wildlife Division of the Department of Lands and Forest^.
The Sanitary Engineering Division maintains a laboratory from which both
field work and analytical work is done.
Recently, increased emphasis has been placed on education, in order th2
excessive pollution caused by a lack of understanding of the harmful effects
of municipal and trade wastes to natural waters would be recognized. Cours
are now provided for sewage disposal plant operators and sanitary inspector
and included in their course of instruction is a section on the life in streams
and the effect of pollution. In addition to these courses, the Pollution Contrc
Board of Ontario has sponsored an Industrial Wastes Conference for the
past two years. Technical papers are presented on waste problems and
treatment, and each year one or more papers have been devoted to pollution
biology.
Probably the most intensive combined biological and chemical inves-
tigation made in Ontario concerned the pollution of the Spanish River. This
study was promptedby litigation and an injunction obtained by riparian land-
owners against a Kraft paper mill. The injunction was dissolved by an act
of legislature in 1950 pending the investigation. The study was made over
the period of one year, during which time continuous observations were mad
on biological conditions and samples were regularly taken for chemical anal
sis. A quantitative study was made of the bottom organisms from above the
mill to the mouth of the river, a distance of about thirty-two miles. The
graphic pattern of numbers and species of bottom dwellers clearly demon-
strated the deleterious effect of the mill effluent to the river.
Nets were fished more or less continuously over the year, and obser-
vations were made on the species and relative abundance of fishes inhabi-
ting the river. It is interesting to note that while fish could traverse the
river until stopped by the mill dam, they did not remain in numbers in the
section extending for twenty miles below the mill. During the period of
investigation, a spill of rosin acid soaps occurred in the mill and passed to
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the river. As this "slug" of toxic material flowed downstream, it killed
all fish in the river and a considerable number for a distance of two miles
into the lake. One beneficial result, however, was the killing of an incredible
number of lamprey ammoecetes.
The changing of the bottom environment by the deposition of fibre was
found to be of prime importance in the elimination of fish food organisms
over an extended section of the river.
An interesting observation from this study was the lack of any chemical
evidence of serious pollution apart from the time of toxic concentrations
of rosin acid soaps .
While this is the only detailed biological study that has been made
of pollution from paper mills, it has provided a valuable basis for future
surveys.
Mining is a second major industry from which pollution problems arise.
Generally, the mines utilize hard rock ores pulverized in many cases to
minus 200 mesh. The settling characteristics of these slimes are poor
and the productivity of some northern lakes are seriously affected by an
artificial turbidity and an unstable bottom. A unique method of determin-
ing the rate of deposition of slimes has been devised by using dust-collector
cans similar to those used in air pollution studies. These cans are set
on the bottom for a specified period of time, then lifted and the contents
filtered, ashed and weighed, thus providing a quantitative estimate of the
deposition of inorganic solids .
Recently, the mining of uranium ores has added a new problem. The
extraction of uranium oxide is made at a pH below one. The highly acid
Mature of the waste and the leaching of toxic metals which are not recovered
make this an especially potent effluent. The problem is further complicated
as the natural waters of the region are slightly acid and poorly buffered.
Natural populations of Salmonoid species of fish will necessitate a high
degree of treatment. This is an example of a knotty problem that may be
solved by the use of bio-assay.
While little use has been made of the practical aspect of bio-assay in
Pollution control, Dr. F. E. J. Fry and his associates of the Ontario Fisheries
Research Laboratories have used it extensively in physiological studies on
fishes .
A study is presently being completed of the cause and control of
excessive blooms of blue-green algae. The field work has been done on
the Kawartha Lakes, a chain of eutrophic lakes in Southern Ontario. The
Merest in this problem began with the death of cattle and other animals
after drinking water containing the algal toxin. Records have been collected
°f at least thirty-six cattle in this region whose death was due to the toxin
*rom blue-green algae.
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The study was divided into three phases:
(1) A study of the biological and chemical conditions in the lake.
(2) A study of control measures .
(3) A study of the toxins produced by the algae.
The following are a few of the observations and findings of this experi-
ment.
The plankton were found to grow in quantity at most times of the year,
with a definite preponderance of blue-greens during the summer months.
The blue-green counts during the four summers that the experiment was
conducted reached peaks between 800,000 and 1,200,000 units per litre.
A number of chemical determinations were made, including total and soluble
phosphorous, various nitrogen analyses, etc. The high productivity of the
lake is connected with the fertility, and phosphorous is believed to be of
prime importance.
In considering control, the one important source where excess plant
nutrients might be removed was the sewage disposal plant for the town of
Lindsay. A study was made of the fertilizing quality of the effluent, and
a laboratory investigation was begun on methods of removing phosphorous
from sewage. Alum, lime, ferric chloride and other chemicals and
combinations were tested. Ultimately a choice of alum and activated silica
was made because of the strength and settling characteristics of the floe.
In addition, the presence of activated silica appeared to enhance the ability
of alum to absorb phosphorous. Continuous treatment was started on a
trial basis in 1954. In May of 1955, treatment began and extended until
the end of September .
The Lindsay disposal plant provides primary clarification only and has a
dry weather flow of 1,200,000 Imperial gallons. While there were no special
facilities for building floe other than the turbulence within the flow, a reason*
good coagulation was obtained and a total of 81% of the total phosphorous
was removed. Soluble phosphates were virtually eliminated, and we are
reasonably sure that most of the 19% remaining in the effluent was lost with
a portion of the alum floe which did not settle. It was estimated that during
the five months' treatment last summer, a total of 8487 pounds of phosphorou
as "F", was removed. If this figure is converted to tons of fertilizer using
superphosphate as an example, it is equivalent to 54 tons. Other improve**1®*
noted were: a 71% reduction in BOD, a 63% removal of suspended solids
and a clear effluent.
The average addition of alum was 94 ppm and silica was 3.4 ppm. The
total expenditure for chemicals was $5006.00, a cost of $25.75 per million
gallons.
It would appear that with proper facilities for mixing and settling,
virtually all the phosphorous could be removed and saving would undoubtedly
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be made on chemicals . This is believed to be the first use of an alum acti-
vated silica treatment and one of the first experiments on continuous phos-
phorous removal from an operating disposal plant.
While a review of the findings has not yet been completed, a considerable
drop was noted in the concentration of phosphorous in the river that receives
the effluent. The soluble phosphorous in the lake was very low and remained
lower than any of the previous years. The total phosphorous was approxi-
mately the same as in previous years and while the blue-green algae built
up rapidly under favourable weather conditions that occurred all summer,
they levelled off at a peak a little less than would have been expected from
previous observations and generally remained at that level for the duration
of the season. While it is unlikely that a definite correlation will be shown,
it is felt that if treatment was continued on a permanent basis, the level of
plant nutrients could be lowered sufficiently to maintain a balanced algal
population and a reduction of the incidence of nuisance blooms .
The investigation into the nature of the toxin developed at the time
of blooms of blue-green algae has been done by the Defence Chemical
Laboratories . While not much information is available yet, toxicity to
rnice has been demonstrated in unialgal cultures. Previous investigations
into the nature of the toxin have shown that it does not belong to any of the
known poison groups. It is hoped that the development of pure cultures,
Much is proceeding at present, will lead to the identification of this toxin.
The work done by the Sanitary Engineering Division is primarily
routine analysis and regulatory in nature in the fields of water supply and
Water pollution. There are, however, projects such as have been previously
Mentioned that the Division is called upon to study or direct. Undoubtedly,
this function will increase in the future as the population and industry grows.
The people of Ontario are justly proud of the abundance of desirable
8pecies of fish in their waters and care is being taken to conserve this
valuable resource.
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REPORT ON POLLUTION STUDIES CONDUCTED IN WESTERN CANADA
Michael Waldichuk
Fisheries Research Board of Canada,
Biological Station, Nanaimo, B.C.
I. Industrial Pollution
Emphasis in pollution studies in British Columbia has been on the indus-
trial development of forest product industries . There are at present eight pulp
mills on the B.C. coast, four of which are undergoing expansion. One new
pulp mill is being constructed, and at least two new pulp mills are in the
embryonic stages of planning. Characteristically large water users, pulp
mills can be a major source of water pollution. The research work has been
closely coordinated with the supervisory branch of the Department of Fisheries,
which receives the requests for approval of plans for waste discharge from
industry. The policy that "prevention is more effective than abatement"
has been continued, and pollution problems are carefully assessed before
they occur. In most instances an effort is made to draw on existing data
and knowledge in these studies, inasmuch as it is not desirable to hamper
industrial development by delays of long-term research. Where additional
survey data or bioassay information are required, these are taken as facili-
ties and manpower permit.
From the point of view of economy and lower pollution, new pulp mills
are installing equipment for production of Kraft (sulphate) pulp as opposed
to the sulphite process. The Kraft process permits reclamation of the salts
in the "black liquor" by evaporation and burning of the organic material.
("Black liquor" is the dark brown fluid which results from the cooking of
wood chips in an alkaline solution at about 330°F under pressure in a diges'tor),
Any of the liquor which escapes in the wash from the screens is generally
too dilute for economical recovery. It is this wash water which constitutes
the major pollution hazard from a Kraft pulp mill. Sulphite digestion is pre-
dominantly used in the older pulp mills . An economical recovery process
reclamation of the salts in sulphite waste liquors has not yet been per-
fected for wide use. In general, effluent from a given tonnage of sulphite
Pulp produces about 10 times as great a pollution hazard as that from an
e(lual tonnage of Kraft pulp.
Newsprint mills contribute a relatively small pollution. The mechanical
binding Of wood in groundwood mills results in a discharge of wastes con-
taminated only by sap, bark extracts, and fibres from the wood. Most of the
dilution stems from the oxygen demand of the organic constituents during
^composition.
The major factors which are responsible for the harmful aspects of
mill wastes to fish are:
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(1) Biochemical oxygen demand
(2) Direct toxicity
(3) Destruction of food organisms
Analysis of pollution problems with respect to Kraft mill effluent has shown
that where dilution is sufficient to combat the B. O. D. {biochemical oxygen
demand) of the waste, direct toxicity is only secondary. Bioassay studies
conducted at this Station show that concentrations greater than 4.0% of Kraft
mill effluent are toxic to young sockeye in sea water for periods greater
than one week (Brett and Alderdice, 1954). No research has been pursued
on the effect of Kraft mill effluent on fish food organisms. There has been
no evidence of a severe reduction in plankton and benthicfauna in regions
of Bubol Columbia where pulp mill wastes have been discharged.
There have been three basic types of marine systems on the B.C.
coast where pulp mill pollution conditions have had to be studied.
A. Inlet-type - Numerous pulp mills in British Columbia are located at
the head of an inlet, often adjacent to an estuary. The choice of such a locality
stems from the availability of good forests nearby, ease of transportation
of logs from inlet logging regions, fresh water supply, hydroelectric power,
and favourable harbour facilities . As a hazard to anadromous fishes these
pulp mills require the greatest precaution in effluent disposal. Wastes passi*1#
into estuarial waters come directly into the path of migrating salmon. A
severe case of pollution might completely wipe out a salmon run.
Evaluation of the capacity of an inlet system to receive wastes according
to oxygen supply and dilution has been based on the study of Tully (1949)
on Alberni Inlet. Waters from runoff undergo a continuous displacement
seaward. The use of the fresh water as a tracer to determine the movement
and mixing of effluents has permitted prediction of pollution with varying
pulp production. Being of about the same density as fresh water, pulp mill
effluent mixes only in the surface brackish layer. Thus the most effective
control on the extent of pollution in an inlet-type condition is (1) controlled
discharge of fresh water above a certain minimum flow, (2) release of the
effluent at the surface and (3) maintenance of the effluent in the jet stream
of the surface flow.
The problem of pulp mill effluent discharge in volumes above the safe
capacity of natural receiving waters was met in the expansion plans of the
Port Alberni pulp mill. Present production of 230 tons of unbleached Kraft
pulp, established on the basis of Tully's (1949) work, does not impose an
excessive pollution load on the inlet. The effluent can be flushed effectively
from the inlet by the natural tidal conditions and Somass River discharge. ^
But the expansion to roughly 500 tons of Kraft pulp and about 500 tons of ne^1
print per day would impose an oxygen demand on the inlet system in excess
of the natural supply. This is particularly true with the reduced river flow
during the late summer. A stream which normally fluctuates from about
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Figure 1.
Chart of the
and proposed
British Columbia coast showing
pulp mill locations«
existing
-------
300 c.f.s. in September to peaks of 30,000 c.f.s„ in November, the Somass
River must be maintained at a discharge of 1,000 c.f.s. or greater to over-
come any serious pollution from the proposed mill.
B. Coastal Seaway type - In this type of location, effluent is discharged
directly into a seaway, use being made of tidal currents and mixing to re-
move and disperse the wastes rapidly. There is seldom any estuary adjacent
to such pulp mill locations so that fisheries of salmon or trout are not likely
to be endagered. At the Harmac (Nanaimo) pulp mill, effluent from a daily
production of 600 tons of fully bleached Kraft pulp is discharged into Northum-
berland Channel. Tidal currents here are predominantly southbound (ebb)
reaching a knot on certain stages of the tide. The geography is such that
little of the effluent reaches the vicinity of Nanaimo Harbour, but is rapidly
funnelled into the turbulent waters of Dodds Narrows. With currents commonly
over 5 knots strength, pulp mill effluent is completely mixed into the sea
water in Dodds Narrows, no traces being detectable below the narrows.
Other examples of this type of effluent discharge can be seen at the
Duncan Bay (Campbell River) pulp mill and at Powell River. Duncan Bay,
located on Discovery Passage, is relieved of its effluent by the strong tidal
currents (up to 7 knots) just south of Seymour Narrows. Powell River dis-
charges effluent into more quiescent waters of Malaspina Strait and Algerine
Passage. No evidence of pollution has been reported in either region.
Popular sport fishing for salmon is especially renowned at Campbell River .
Any pollution which could arise in these cases in future expansion would
probably result from an adverse effect on migrating juvenile salmon. In
Discovery Passage large schools of young salmon seek the shelter of the
bays and inlets in their journey to the sea. It is hoped that observations
can be made on the effect of expansion of the Duncan Bay pulp mill on the
young salmon passing through the bay during the summer.
C. Restricted Embayment type - Being intermediate between the inlet-
type condition and the coastal seaway, the restricted embayment does permit
some flushing by virtue of its openness to an adjoining channel. Generally,
however, it possesses a circulation all its own independent of that in adjoin-
ing channels. Hence a certain amount of stagnation results.
The pulp mill at Port Edward near Prince Rupert is an example of this
type of situation. Shallow, restricted conditions in Wainwright Basin and
Porpoise Harbour hinder the flushing of the effluent into Chatham Sound.
Large concentrations of mill wastes have been particularly evident during
low tides . Mixed reports have been received on the effect of the wastes on
the biotic environment. Some observers have reiterated that all marine
life in that vicinity is being destroyed. But observations carried out by
biologists of the Department of Fisheries (May, 1953) cl^im that these re-
ports are exaggerated, and that both fish and bottom fauna appear to pre-
vail in normal health. A further investigation of the region is planned in
the near future from the Biological Station, Nanaimo.
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The proposed pulp mill at Crofton will create another sample of a
pulp mill in a restricted embayment, Osborn Bay, into which the
effluent would be normally discharged, is relatively sheltered from
Stuart Channel; its flushing and circulation are sluggish. The bay
is fringed, particularly at the northern end, by oyster-growing leases.
Living primarily in the intertidal zone, oysters are vulnerable to any
harmful wastes found in the surface waters. In order to prevent the
lost of oysters entirely, action would have to be taken to discharge
the effluent outside the oyster-growing area or render it innocuous
by treatment. The most satisfactory solution to industry economically
was the piping of mill wastes into deep water (10 fathoms) in Stuart
Channel beyond the Shoal Islands protecting Osborn Bay. Recommendations
have been made to this effect and have been met with agreement by
both the Provinical Department of Fisheries, Federal Department of
Fisheries and Industry.
II. Domestic Pollution
A limited amount of work has been conducted on pollution from domestic
wastes in coastal communities of British Columbia.
A. Vancouver Sewage Disposal
Study of the effects of sewage disposal from Vancouver was undertaken
as a cooperative effort by the Pacific Oceanographic Group, National
Research Council, Institute of Oceanography of the University of British
Columbia, Tidal Branch of the Hydrographic Service of Canada, and Air
Surveys Branch of the British Columbia Department of Lands and Forests
at the request of the Vancouver and Districts Joint Sewerage and Drain-
age Board. Oceanographic data were collected seasonally in the Fraser
River estuary and in Vancouver Harbour over a period from September,
1949 to March, 1951.
Preliminary analyses of these data along with tidal current information
and aerial photographic surveys have been used for establishing the circula-
tion in Vancouver Harbour and at the Fraser River estuary. These studies
showed how the tides and discharge from the Fraser River combine to
produce a characteristic circulation in Burrard Inlet.
Off Point Grey, there is a predominent northward set of the current
into English Bay on all stages of the tide. Off Point Atkinson, at the
northern shore of the entrance to Burrard Inlet, there is a prevailing
westward current out of English Bay. A counterclockwise main eddy
system with numerous smaller eddies and stagnation points superimposed
on the principal circulation were noted to exist in English Bay.
The results of the oceanographic studies have permitted the evaluation
of different outfall points for their suitability in sewage disposal. Final
choice of discharge locations was based largely on the effectiveness with
which the sewage would be dispersed and removed by the currents.
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B. Nanaimo Harbour Sewage Disposal
An oceanographic study of currents in Departure Bay and Naiiaimo
Harbour was conducted as a cooperative venture between the Pacific Oceano-
graphic Group and community groups of the city of Nanaimo. A large measure
of assistance was given by the Nanaimo Yacht Club with their boats , and much
volunteer help came from the Nanaimo city employees. Close cooperation
was maintained with the local branch (Central Vancouver Island Health Unit)
of the Provincial Department of Health and Welfare. The prime concern in
the survey was the determination of existing pollution from a health stand-
point and further contamination of beaches to be expected from additional
sewage discharge.
Results of the survey indicated that Nanaimo Harbour is already polluted
by existing sewer outfalls and that beaches within the harbour are not fit to
be used for bathing. Departure Bay beach is still free from contamination,
but is threatened by any sewage discharged at Brechin Point (located at the
northern end of Newcastle Island Channel between Nanaimo Harbour and
Departure Bay). Surface currents in both bays are largely wind-driven.
The prevailing southeasterly winds would drive any increased sewage out-
put into Departure Bay and gradually wipe out the remaining beach recrea-
tional area in the district.
Recommendations were submitted to the City Council of Nanaimo that
either the sewage be totally treated before discharge, or it should be piped
beyond the barrier islands (Newcastle and Protection Islands). These re-
commendations are being acted on at present and further survey work is
awaited to determine the most appropriate discharge points outside the islands-
-------
REFERENCES
Brett, J. R. and D. F. Alderdice. MS. Bioassay. Studies with reference
to Kraft Mill Effluent in Alberni Inlet.
Tully, J. p. 1948. Pollution research in Alberni Inlet. Fish Res. Bd.
Canada, Pacific Prog. Rep. No. 76, pp. 66-71.
_ , 1949. Oceanography and prediction of pulp mill pollution in
"" Alberni Inlet. Bull. Fish. Res. Bd. Canada, No. 83, 169 pp.
Waldichuk, Michael and J. P. Tully. 1953. Pollution study in Nanairno
Harbout. Fish. Res. Bd. Canada, Pacific Prog. Rep. No. 97, pp. 14-17.
Waldichuk, Michael. 1954. Effect of pulp mill waste in Alberni Harbour. Fish.
Res. Bd. Canada, Pacific Prog. Rep. No. 101, pp. 23-26.
, 1955. Effluent disposal from the proposed pulp mill at
Croftori, B. C. Fish. Res. Bd. Canada, Pacific Prog. Rep. No. 102,
pp. 6-9.
. 1956. Pulp mill pollution in Alberni Harbour, British
Columbia. Sewage and Industrial Wastes, 28(2):199-205 .
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The Relationship of the Polychaetous Annelid Capitella capitata
(Fabricius) to Waste Discharges of Biological Origin
Donald J. Reish
Department of Biology and the Allan Hancock Foundation,
University of Southern California
INTRODUCTION
Biologists have long recognized the shortcomings of chemical and
physical measurements of water quality, and thus have searched for organisn
which could serve as indicators of different degrees of pollution. Organisms
particularly those that are attached or bottom inhabitants, are favored by
many since they reflect the water conditions not only at the time of sampling
but for some time previously.
The use of organisms as indicators of pollution in marine waters has
lagged considerably behind that of fresh water studies. Wilhelmi (1916)
mentioned that the polychaete Capitella capitata played a similar role in
marine waters as Tubifex does in the fresh waters of Germany. Blegvad
(1932) studied the bottom fauna in the vicinity of domestic outfall sewers
in Copenhagen Harbor, Denmark. He was able to divide the region surround-
ing one outfall into three zones; an inner region lacking animals and with
'he substrate characterized by a sulfide odor, an intermediate zone con-
taining a few animal species and with the substrate possessing a sulfide odor,
and a third zone showing no measureable effects of the discharge. A second
domestic outfall lacked the intermediate or marginal zone. Filice (1954),
forking in the Castro Creek area of San Francisco Bay, also separated
the bottom fauna into three zones; (I) a healthy zone unaffected by waste
discharges, (2) a marginal zone characterized by a few tolerant species,
Notably the polychaetes Capitella capitata, Neanthes succinea (Frey and
Leuckart), and Streblospio benecTicti Webster, and the molluscans Mya
^renaria Linnaeus and Macoma inconspicua (Broderip and SoWerhY), and
T3) a zone essentially lacking in animals. Recently the author (1955) pub-
lished the general results of three quantitative bottom surveys conducted in
the Los Angeles -- Long Beach Harbors. This area, which receives waste
discharges of domestic and industrial origin (Anon, 1952), was divided into
five zones on the basis of bottom conditions. Capitella capitata was found
to be particularly abundant in regions receiving effluents of biological origin,
^his organism was the characteristic animal in what was termed the polluted
bottom zone. The present report discusses some of the results of bottom
®urveys made in the Los Angeles-Long Beach Harbor and in some other
Marine waters of southern California. Emphasis has been placed upon the
°ccurrence of C. capitata and its possible role as an indicator of pollution
biological origin. Biological wastes are herein defined as discharges
frOm fish canneries, domestic sewage, and garbage.
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OBSERVATIONS
Los Angeles -- Long Beach Harbors (Figure 1). The bottom of the
Los Angeles -- Long Beach Harbor has been sampled five times during
the past five years. Three of the samplings were made in 1954. The dis-
tribution of C. capitata for the June 1954 Survey is shown in Figure 1.
Slip 5 of LosTAngeles Inner Harbor receives waste discharges of domestic
and fishery origin. The bottom of the inner portion of the slip was covered
with fish scales . Capitella capitata was found in the area in large numbers
along with a few other species of invertebrates. Domestic, fishery, and non-
biological industrial wastes are emptied into Slip 2 of Long Beach Harbor.
Here this worm was found with one other species of polychaete. The region
of fish harbor in the Los Angeles Harbor receives some of the effluents
from the fish canneries and the bottom was covered with fish scales. Ca-
pitella capitata was collected either alone or with one or two other species
of polychaetes.
Following primary treatment the effluent of the terminal Island sewage
treatment plant is discharged into the Los Angeles Outer Harbor. Three
outfalls from the fish harbor canneries are located nearby (Figure 2).
The samples taken in 1954 at the sewage outfall either contained C. capitata
or lacked animals. Additional stations were sampled in December~l955
(Figure 2) to determine whether or not an intermediate assemblage of
animals existed between the polluted zone possessing C. capitata and the
healthy zones typical of the outer harbors (Reish, 195"E>Tsee also Hartman,
1955, Stations 29 and 44b). No animals were taken from the samples near
the sewage or fish cannery outfalls. The substrate possessed a strong
odor of domestic sewage. Capitella capitata was found in the stations a
short distance from these outfalls, The farther the stations were located
from the outfall the more varied the fauna became. There was no indication
of an intermediate zone between the polluted bottom containing C. capitata
and the healthy area.
Alamitos Bay. This bay, which is located at the southern boundary of
Los Angeles County, is essentially a clean body of water used primarily
for recreation (Reish and Winter, 1954). Capitella capitata was found at
three stations in the vicinity of a public dump where garbage was pushed
into the bay. This species was present at two other widely separated stations
where there was no known pollution. There two stations were the only ones
sampled where C. capitata had been collected where there was no pollution
of biological origin discharged nearby.
Lower San Gabriel River. The tidal portion of this river is separated
from Alamitos Bay by a jetty. This region was sampled in 1952 and again
in 1954 (Reish8 1956). No animals were found in the bottom materials in
1952, but animals were encountered at some of the stations in 1954. A
portion of the river was diked and dredged about four to five feet deeper
in 1952 after the survey of that year. This dredging activity removed the
accumulated sludge from the river bottom. A total of 12 different species
was collected in 1954, of which C. capitata was the most common. This
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FIGURE I
-------
BOTTOM CONDITIONS DECEMBER 1955
HEALTHY BOTTOM
* 1' Jb POLLUTED BOTTOM
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polychaete was found near the outfall sewers of the City of Seal Beach and
downstream from the Los Alamitos Naval Air Station.
Newport Bay. This body of water is also essentially clean and used
primarily for recreation. However, C. capitata was taken along with fish
scales at one station in 1951 and again in 1954. This station was situated at
the end of a small arm of the bay in the proximity of fish canneries.
DISCUSSION
Gaufin and Tarzwell (1952), while concerned with fresh water pollution,
included four points that should be considered when testing possible indicator
of pollution. These criteria are; (1) large number of individuals, (2) few
species in the fauna, (3) principally scavenger feeding habits, and (4) either
« toleration for low dissolved oxygen or possess some adaptation to a low
dissolved oxygen environment. At least some of these qualifications are
fulfilled by C. capitata. It has been encountered in large numbers, as many
58 400 haviqgljeen taken in a sample covering a surface area of 100 square
inches. Sometimes it was the only species taken. More frequently there
^ere a few additional forms present In those samples containing organisms
^nly three or four species other than C. capitata were observed at the sta-
tions nearest the Terminal Island sewage treatment plant and the fish canner-
Outfalls. In contrast, there were 15 or more species present at each station
•hroughout much of Los Angeles -- Long Beach Outer Harbors. Capitella
Capitata burrows into the substrate and engulfs substrate in much the same
banner as an earthworm. The oxygen requirements of this worm have
*ot been studied but it has been encountered in bottoms in Los Angeles Inner
^arbor where the overlying water lacked oxygen at the time of sampling,
however, this was exceptional, A more typical situation was 3.5 ppm oxygen
the stations where C. capitata was taken.
It is not known whether or not a relationship exists between the number
C. capitata present in a sample and the degree of the biological pollution.
^ieTcT "dataand laboratory observations indicate that C capita^ has %. short
history as it reaches sexual maturity in about a month. It is capable
reproducing throughout the year in southern Californian waters. It is not
lI*own whether or not any reproductive peaks occur during the year. This
'Prices is cosmopolitan in its distribution,, but, with the exception of the
ltatement by Wilhelmi (1916), the ecology of C ¦ capitata has not been studied
n the other geographical areas
SUMMARY
The use of the polychaete
Possible indicator of pollution of biological origin
tu- - t. u -nrmmtered near outfalls discharging biological
This species has been enc°""t*" h Harbors, Alamitos Bay, Lower
Pastes in Los Angeles -- Long Beacft w
San Gabriel River, and Newport Bay.
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3, A special study made in the vicinity of sewage and fish cannery
outfalls in Los Angeles Outer Harbor showed no intermediate
assemblage of animals between the polluted C. capitata zone and
the healthy zone characteristic of much of the outer harbor.
Research Grant No. RG-4375-C2 from the National Microbiological
Institute of the National Institutes of Health, U. S. Public Health
Service, supported the subject investigation.
LITERATURE CITED
Anon, 1952. Los Angeles -- Long Beach Harbors pollution survey.
Calif. , Los Angeles Regional Water Pollution Control Board No.
4. 43 pp.
Blegvad, H. , 1932. Investigations of the bottom fauna at outfalls of
drains in the Sound. Rept. Danish Biol. Station. 32: 1-20.
Gaufin, A. R. , and C. M. Tarzwell. 1952. Aquatic Invertebrates as
indicators of stream pollution. Public Health Reports. 67:57-64.
Hartman, O. , 1955. Quantitative survey of the benthos of San Pedro
Basin, California. Allan Hancock Pacific Expeditions. 19: 1-185.
Reish, D. J. , 1955. The relation of polychaetous annelids to harbor
pollution. Public Health Reports. 70: 1168-1174.
1956. An ecological study of Lower San Gabriel River, California,
with special reference to pollution. Calif. Fish and Game. 42:51-61-
and H. A. Winter, 1954. The ecology of Alamitos Bay, California,
with emphasis upon pollution. Calif. Fish and Game. 40: 105-121.
II M
Wilhelmi, J. , 1916. Ubersicht uber die biologische Beurteilung des
Wassers. Ges. naturf. Freunde Berlin, Sitzber. vol. for 1916,
pp. 297-306.
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COOPERATIVE RESEARCH AT OREGON STATE COLLEGE IN
THE BIOLOGICAL ASPECTS OF WATER POLLUTION V
Charles E. Warren
Oregon State College
Department of Fish and Game Management
and
Peter Doudoroff
U. S. Public Health Service
and Oregon State College
Corvallis, Oregon
Water pollution problems are essentially limnological or oceanographic
field problems. Yet, the variability and complex interactions characteristic
of real situations make the understanding of that which is observed in the field
so difficult that a considerable part of water pollution investigation has had
to be carried on in the laboratory. Idealization of the laboratory experiment
greatly facilitates the analysis of results, but the idealized experiment rarely
approximates reality. Field problems being often too complex for thorough
analysis, and laboratory models being usually too simple to be truly represen-
tative of natural situations, neither field investigation nor experimentation
in the laboratory alone is sufficient for the solution of many water pollution pro-
blems. The best information that can be obtained by each approach is needed,
and it is highly desirable to bridge the wide gap between ordinary field obser-
vations and pertinent idealized experiments. The more nearly laboratory
experiments can be designed to model actual situations under study and
still retain the advantageous feature of comparative simplicity, and the more
nearly a field study can be made to resemble an idealized experiment through
the control of variables, the more efficient will be the investigation and the
more reliable the interpretation of the findings. Many current problems
having to do with the biological aspects of water pollution can be solved only
by utilizing these different approaches in an entirely complementary manner.
The cooperative program of investigation in water pollution biology
being conducted at Oregon State College by the Department of Fish and Game
Management and the R. A.Taft Sanitary Engineering Center of the U.S.
Public Health Service includes such complementary field and laboratory
research. Personnel of these two organizations, with the aid of a number
of graduate students, are participating in a research program designed to
provide fundamental information pertaining to present and future biological
Problems of fresh-water and marine pollution. Close cooperation with various
state regulatory agencies and with industry is helpful in keeping this research
Pertinent to present-day problems and also facilitates anticipation of future
Problems .
Current field studies and those soon to be initiated at Oregon State College
fall into two general categories. The first of these categories includes broad
studies of the physics, chemistry and biology of rivers and streams variously
affected by pollution. Through these studies it is hoped to increase available
information 011 the influence of domestic and industrial wastes on fresh waters,
*/ Miscellaneous Paper No. 29, Oregon Agricultural Experiment Station
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to shed more light on the value of biological indicators, and to furnish ideas
and direction for the over-all research program. The second category in-
cludes those studies which are more nearly field experiments than they are
field surveys . Small streams which can be purposely subjected to controlled
experimental pollution and variously modified as necessary, and rivers
where appropriate experimental designs of a statistical nature can furnish
a mathematical control are to be utilized for these studies.
Plans have been made to undertake research on a small stream in which
the desired dilutions of added chemicals or wastes can be maintained. This
field study will aid in extending information resulting from laboratory studies
in which a system of artificial streams discussed below is employed. Field
observations and experiments under the somewhat controlled conditions
attainable in a small stream will make possible detailed investigation into
the influences of wastes on the productivity of such streams at the plant,
herbivore, and carnivore levels of production, the last level including game
fish.
The results of short-term laboratory experiments designed to determine
what conditions are rapidly lethal for aquatic organisms held in glass vessels,
though informative, are of limited practical value. In order to infer from
such ordinary bioassay results what changes of water quality can be tolerated
for long periods by an organism in its native environment, it must be assume**
that the manner of action of the lethal agent under consideration is the same
at rapidly fatal and at slowly fatal concentration or intensity levels, and the
same in the natural environment as in the aquarium. This assumption is
by no means always a valid one. Furthermore, though environmental conditio*1®
may not be so adverse as to be demonstrably fatal to aquatic organisms,
their effect on a population of these organisms may still be thoroughly
destructive if they interfere with the reproduction, development, feeding,
growth, normal activity, or migratory movements of individuals of that
population. Chronic injury to fish populations, due to pollution, may well
be much more common and important than spectacular mortialities of fishes
caused by acutely harmful pollutional conditions of relatively brief duration,
which may or may not have a serious and lasting adverse effect on fisheries.
Therefore, some laboratory experiments should be designed so that their
conditions approximate selected features of the natural environment. Some
of these experiments should be of prolonged duration and should measure the
effects of the tested conditions on some of the essential life processes and
over-all well-being of organisms, attention being given to the most susceptib*®
stages of the life-history of the organisms . Knowledge of the concentra-
tion of a toxic waste, or of dissolved oxygen, which can be barely tolerated
for a short period of time, or when the organisms may be relatively resistant
to adverse conditions, may be necessary, but is not sufficient.
Encouraging results have been obtained at Oregon State College by using
wooden troughs with various current and bottom conditions as artificial
streams for the purpose of evaluating the effects of exposure to relatively
low waste concentrations for periods as long as a month on different aquatic
organisms. In this way, it has been possible not only to determine long-ten**
lethal concentrations of waste, but also to note the effects of the waste on
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the feeding and growth of fish, on the development, habits , and emergence
of insects and on other life processes of aquatic organisms, such as ecdysis
in young crayfish. Changing bottom conditions such as the excessive production
of periphyton can be closely observed, as can be the influence of the periphyton
on bottom-dwelling forms. These experiments not only make possible a deter-
mination of the concentrations of a waste having marked effects on the or-
ganisms under artificial stream conditions , but may suggest some of the
reasons for these effects .
Thus , when stonefly and caddisfly larvae were held in troughs receiving
pulp-mill wastes in various concentrations, it was possible to observe closely
the environmental conditions under which mortality occurred and the condition
and behavior of the animals before death. The abundant growth of periphyton
over the rocks placed on the bottom, and also on some of the experimental
animals, the upward movement of the insects from the undersides of rocks
where they are usually found (which, in nature, could make them more sub-
ject to predation), and the changes of dissolved oxygen concentration among
the rocks and beneath the blanket of periphyton all could be readily noted
or measured Furthermore, in seeking to determine the causes of distress
and mortality of the insects (which may be referable to toxicity of the wastes,
to oxygen deficiency, to some mechanical effect of the periphyton, or to
their combined influence) it has been possible to evaluate the role of a single
environmental factor by modifying the artificial environment with respect
to that factor. For example, by introducing oxygen into the water before
it is mixed with waste and enters the troughs, relatively high dissolved
oxygen concentrations have been maintained, compensating for the oxygen
demand of the waste. In this way, and through additional experiments on the
influence of dissolved oxygen concentration and current velocity on the
survival of insects in cages inserted in glass tubes, which also have been
undertaken, the part played by dissolved oxygen reduction in causing the
observed pollutional damage to these aquatic animals can be determined.
Long-term experiments with complex industrial wastes inevitably
present many problems . One of these is the variability among the several
waste samples or batches necessary for completing a single experiment,
large amounts of waste being required. The same volumetric dilutions of
different waste samples from the same source often differ greatly in toxicity,
so that the analysis of test results obtained without standardization of waste
toxicity would be difficult, if not impossible. Frequently, the toxic com-
ponents of complex wastes are unknown, or there are no chemical or physical
means for measuring and standardizing the lethal factors. Biological standard-
ization of waste samples has been accomplished with apparently good success
by determining for each sample the 24-hour median tolerance limit of one
of the test animals (a fish) being used in the long-term experiment. Some
constant percentage of this median tolerance limit is then the strength of
diluted waste maintained in each experimental trough during the entire course
of the experiment, the dilution used thus being adjusted to the relative acute
toxicity of the individual samples . This procedure has two distinct advan-
tages. First, the toxicity of the trough dilution, at least for the control
species, should remain constant from sample to sample if the short-term
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and long-term effects of the waste do not vary independently. Secondly,
acute lethality data are provided for later comparison with the results of
long-term tests of lower concentrations .
Once the relationships between concentrations lethal to a representative
fish in a short period of time (median tolerance limits) and concentrations
harmful to a variety of stream organisms over a long period are known,
it may be possible to predict the long-term harmful concentrations on the
basis of short-term test results. Sufficient investigation of the toxicity
of a particular industrial waste usually should make possible the determina-
tion of dilution factors which, when applied to the short-term median toleranc<
limit, would yield a reliable estimate of the maximum safe concentration of
that waste in the environment of fish and other organisms of importance as
fish food. An industry, when supplied with these bioassay application factors,
could control waste discharges through routine bioassays of the effluent.
Such bioassays would be no more difficult than many chemical and physical
determinations now routinely used in the control of industrial effluents .
They would, however, provide much greater assurance that the aquatic re-
sources supposedly protected by the waste-control measures are in fact
being protected. Only too frequently, ineffective, though complex, chemical
tests are being used to evaluate the potential toxicity of industrial wastes to
aquatic organisms .
Some of the species which have been used in these experiments for stand-
ardization of wastes are believed to vary in their tolerance to certain adverse
conditions with such variables as size, age, season, source, time in captivity,
and diet. Consequently, the standardization procedure may result sometimes
in adjustments to variation in the standard animal rather than adjustments
to variability among the waste samples . Needless to say, variations in the
tolerance of the fish, as well as variations in the toxicity of the waste, are
of considerable interest in connection with practical application of the results
In order to make possible their separation and study, as well as to provide
a dependable standardization procedure, guppies are now being raised under
very constant conditions to furnish an animal more standard than the wild
fish. Genetic strain, age and state of sexual development of the guppies
at the time of use, as well as the conditions under which they are reared,
such as light, temperature, and diet, can be kept fairly constant; and this
will result, it is believed, in sufficient uniformity of the test animals .
Standardization and control experiments with the guppy will not replace
experiments with species of economic importance (e.g., juvenile salmon)
but will supplement these experiments . Only in this way can variability in
the valuable native fish species be distinguished from variability in the
waste.
A rather broad study entitled "The Influence of Dissolved Oxygen upon
the Survival, Development, Growth, Activity, and Movements of Fresh-Wate*
Fishes" is now being carried on at Oregon State College. The survival of
fish at low concentrations of dissolved oxygen in different waters has been
studied intensively, while the temperature, carbon dioxide content, alkalinity'
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pH and other properties of the water have been varied. In most of these
experiments the test water has been renewed continually. The dissolved
oxygen content of the flowing water is reduced to the desired level by the
controlled bubbling of nitrogen through it while it flows continuously downward
through a glass column. Although the duration of most of these experiments
has been one to five days, some have been continued for as long as thirty
days.
The results of long-term experiments still in progress indicate that
the food consumption and growth rate of salmonid fishes can be influenced
by reduced dissolved oxygen concentrations which are well above the lethal
levels. In these experiments an effort is being made to supply the fish
with a diet approximating a natural diet. The rate of food consumption and
the relative efficiency of its utilization at each of several different dissolved
oxygen levels which are above the lethal level are being determined. It is
planned eventually to investigate also the influence of fluctuating dissolved
oxygen concentrations upon feeding and growth.
Studies of the influence of dissolved oxygen concentration on the rate
of development of salmonid eggs, the percentage of successful hatching, and
the survival of hatched larvae have yielded results of considerable interest.
It appears that, in almost still water at least, the oxygen concentrations
required for successful development and hatching may be quite high in
relation to the minimum levels tolerated by fully developed juvenile fish.
Inasmuch as current must have an important influence upon the minimum
dissolved oxygen requirements of developing eggs, its role needs thorough
investigation in connection with further studies of the oxygen requirements
of the eggs. Studies in the field may be necessary for determining the range
of natural conditions in salmonid redds, so that experimental conditions
can be selected accordingly and the results related to circumstances found
in nature.
It is strikingly apparent that fish which survive in bottles at barely
tolerable dissolved oxygen concentrations are so sluggish or inactive that
they cound not maintain themselves and survive indefinitely in their
natural environment, where they must actively resist currents, feed, and
escape their enemies. With newly developed apparatus, it is possible to
study the ability of fish to resist currents of moderate velocity when they
are in water with any desired dissolved oxygen concentration. The activity
potential of the fish thus can be related to the oxygen concentration.
Preliminary results indicate surprising ability of some fish to resist moderate
currents for long periods at dissolved oxygen concentrations not very far
above the minimum levels tolerated by resting fish. Field studies are
needed, in connection with these laboratory studies, for determining the
velocity of currents that these fish must normally resist for long and short
periods of time in their natural habitats.
The movements of fish, as influenced by variations of the quality of
water encountered, often may determine whether or not the fish will be
exposed to avoidable injurious environmental conditions, and whether or
not they will occur in certain environments where water-quality conditions
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are tolerable. It is known that not all harmful conditions are readily avoided
by fish, and tolerable conditions apparently can be repellent. Avoidance
reactions of fishes to reduced dissolved oxygen concentrations, as well as
to varying dilutions of industrial wastes, are being investigated at Oregon
State College, chiefly through laboratory studies. The application of the re-
sults of the laboratory tests to problems encountered in the field presents
serious difficulties, the circumstances within the confines of even a large
laboratory apparatus being a very poor model of conditions in stream en-
vironments of much greater area, but the tests can nevertheless be instruc-
tive .
In the Pacific Northwest, as in other parts of the country, suitable indus-
trial sites with adequate process and waste-disposal waters are becoming
increasingly scarce. Many industries are now selecting locations adjacent
to marine or estuarine waters . The aquatic resources of many of these
areas are of tremendous commercial and recreational value. Yet, we now
know even less about the basic water-quality requirements of marine organist*1®
and their relative resistance to harmful pollutants than we do about the require*
ments of fresh-water forms. At the marine laboratory of the Department of
Fish and Game Management, studies of the water-quality requirements of
a number of marine forms have been initiated and are to be greatly expanded
in an effort to fill to some extent this serious gap in our knowledge. These
studies are of both short and long duration and include an attempt to reproduce
marine environments in the laboratory. Sufficient field work will be carried
on to assure pertinence to present and fu,ture practical problems .
A research program such as that considered above requires certain
facilities and a location where different species are readily available and
where the desired field studies are possible. The Department of Fish and
Game Management has a fisheries research laboratory on Mary's River at
Corvallis, and a marine research laboratory near Newport;on Yaquina Bay
These laboratories are equipped and operated jointly with the U. S. Public
Health Service. The Corvallis laboratory is housed in five buildings, has a
supply of river water, and six 250-gallon tanks provided with running water
for holding stocks of fish for experimental purposes. Two constant-tempera-
ture rooms (one very large and one small) are available at the Corvallis
laboratory for standing-water experiments. These have both heating and
cooling units . The Yaquina Bay laboratory has both fresh-water and salt-
water systems, a spring furnishing a good flow of fresh water for experiment8
requiring water of considerable purity. Dock and live-box facilities are ade-
quate. There is one large constant-temperature room available at this
laboratory. Yaquina Bay probably has a greater variety of commercial and
non-commercial fish and shellfish and other marine invertebrates than any
other Oregon bay. Corvallis is situated on the Willamette River. This river
system, with its varied stream environments, its wide representation of the
cold-water and warm-water fish species of North America, and its example®
of certain of the effects of domestic and industrial waste disposal, offers
excellent opportunities for field study and is a good source of experimental
material.
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Specialized apparatus has been requisite to much of the work outlined
above. Preliminary experiments with artificial streams as a means of
studying the influences of wastes on stream ecology have proved very en-
couraging. The most recently installed apparatus provides a system of six
artificial streams. Each stream consists of two 10-foot troughs, one having
the water recirculated by a ^-horsepower pump so as to provide riffle-like
currents, the other representing a pool-like environment. The water in
the troughs is continually renewed, flowing river water. Appropriate bottom
materials and lighting result in what is believed to be a rather good model
of a stream environment. After the desired plant and animal communities
have been established, the wastes to be studied can be introduced by means
of chemical pumps in different amounts into the six streams, so as to determine
effects of different waste concentrations.
Another apparatus now in use in the laboratory was devised to make
possible the study of the effects of low concentrations of dissolved oxygen
and other water quality conditions on fish swimming against currents with
velocities up to 1 foot per second. This apparatus consists of a glass pipe
of 4-inch diameter through which the water is recirculated by means of a
centrifugal pump. Water quality in the glass pipe is controlled by exchange
at a rate of about 1 liter per minute, the exchange water having its character-
istics such as temperature and dissolved gas content adjusted by other
appropriate components of the apparatus. An apparatus of this kind now being
constructed should make possible the study of the influence of critical water
conditions on fish resisting currents of relatively high velocities.
The avoidance reactions of fish to water having various characteristics
have been studied in a 2 by 9 foot tank with one-third of its length subdivided by
partitions into four channels. Each channel is equipped with an adjustable
water input and an adjustable drain which, with proper balancing of flows,
result in quite sharp boundaries between waters of different quality at the
channel openings. Other apparatus has made possible the study over short
or long periods of time of the lethal and other effects of water having tempera-
ture, oxygen concentrations, carbon dioxide concentrations, total alkalinity
and waste concentration controlled. A number of such constant-flow experi-
mental units, each consisting of about five test vessels (usually of 12-gallon
capacity) and such other components as are necessary for adjusting the
dissolved gas content or other properties of the water in each container inde-
pendently, are available at the Corvallis and Yaquina Bay laboratories for
studies of the influence of water quality variations upon the survival and growth
of fish and the development of their eggs. A laboratory shop and the coopera-
tion of research apparatus specialists have greatly facilitated the development
°f these and other pieces of equipment necessary for pursuing the problems
under investigation.
The provision of these facilities has been possible only through the
Pooling of resources by the Department of Fish and Game Management and
the U. S. Public Health Service in the joint undertaking of all of these inves-
tigations . Other departments of Oregon State College and industrial repre-
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sentatives participate in many of these investigations. Considerable support
is given these research projects through grants by Federal agencies and
by industry, notably The National Council for Stream Improvement,
Much of this research is accomplished with the aid of graduate research
assistants and fellows who are employed to work on problems suitable for
graduate theses. Master of Science and Doctor of Philosophy degrees may
be pursued with majors in fisheries and minors in desired fields . Several
of these assistantships and fellowships are available each year. The researc
program is complemented by an appropriate instructional program designed
to prepare specialists for employment in the water pollution field. College
and Public Health Service personnel cooperate in the instructional program.
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SOME ASPECTS OF WATER POLLUTION IN THE MISSOURI BASIN
by
Joe K. Neel
U. S. Public Health Service
Kansas City, Missouri
DESCRIPTION
The Missouri Basin (Figure 1) comprises the larger part of arid and
semi-arid regions contributory to the Mississippi Drainage System. Pre-
cipitation declines from about 40 inches per year in the eastern part of the
basin in Missouri to 10 to 15 inches per year along its western boundary
in Montana, Wyoming, and Colorado. The middle basin (the Dakotas,
Nebraska, and Kansas) has annual precipitation ranging from 15 to 30
inches, Long dry seasons are prevalent over most of the basin; and the
river normally exhibits only two high stages each year-- one coming from
melting snow on the praires in March and April, and the other arising
from prairie rains and snow melt in mountainous headwater areas in June.
The main stem and many tributaries are undergoing development for
irrigation, power production, water supply, navigation, flood control,
and waterfowl refuges. The main stem reservoir system, which will con-
sist of three huge one large, and two small impoundments, now lacks
only Oahe and Big Bend Reservoirs. Construction of Oahe is in progress.
Total storage capacity for this reservoir system exceeds 70, 000, 000
acre feet. Reservoirs now in operation (Ft. Peck, Garrison, Ft. Ran-
dall, and Gavins Point) have permitted significant flow regulation and
are retaining a large amount of the suspended sediment carried in
the traditionally muddy river.
Tributary developments are, at present, more concerned with irriga-
tion than the main stem impoundments; although the latter will figure in
development of new irrigation areas. Irrigation water use may involve
natural flow diversion as now practiced in the Yellowstone Valley,
reservoir storage as exists on the North Platte River, or ground water
pumping from river valleys as is customary along the lower Platte
River. Some irrigation reservoirs also furnish hydro-electric power
and municipal water supplies. Impoundments used solely as waterfowl
refuges compose a very small percentage of impounded waters in the
basin. Fish and game interests, however, receive due consideration in
the operation of larger multiple purpose reservoirs, some of which also
3erve as water fowl refuges.
SOURCES OF POLLUTION
The Missouri Basin in primarily an agricultural region, and major
ndustries are concerned with processing agricultural products. Meat
jacking houses are scattered over the basin but their greatest con-
:entration, and greatest waste contribution is along the Missouri River
torn Sioux City, Iowa, to Kansas City, Missouri. Wastes from these
istablishments reach the river in a raw state.
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The beet sugar industry, whose refineries or factories discharge
a large seasonal waste load, is largely confined to irrigated areas along
tributary streams, e.g., the Platte, Big Horn, Belle Fourche, and
Yellowstone Rivers. Oil fields and petroleum refineries are located in
several areas in the Missouri Basin. The greatest concentration of
refineries is in the Kansas City area, but significant contributions of
these wastes affect various streams, the North Platte, Yellowstone,
and others, Other industrial wastes--steel mill, distillery, and chemical
processing discharges--enter the river at the larger main stem
municipalities. Salt brines, originating in natural deposits and supple-
mented by oil well waste flows, are a problem in the Smoky Hill,
Solomon, and Saline Rivers in the Kansas Basin.
Missouri Basin streams receive considerable quantities of domestic
and municipal wastes, largest concentrations being poured into lower
reaches of the main stem. Most municipalities situated on the Missouri
lack any form of sewage treatment and raw domestic wastes considerably
augment those from industries at the larger cities. Some tributaries are
superior to the main stem in this regard. Major municipalities on the
James River, for example, all provide wastes reduction equivalent to
conventional secondary sewage treatment. Some towns on other tributaric
provide treatment for domestic wastes, but allow certain industries to
discharge wastes in the rawest form. Billings, Montana, for instance,
has a municipal sewage plant, yet major industries--oil refineries, beet
sugar plants, and packing houses--provide no waste treatment.
Badlands, regions of easily erodable materials lying above local
base levels, are characteristic of more arid regions in the Missouri Basi
Their existence depends upon paucity of rainfall, as those rains that fall
bring great quantities of such materials into the Big Horn, Powder, Yello
stone, Little Missouri, and Missouri Rivers. Badlands contribute the
largest share of the silt load borne by the Missouri, and were mainly
responsible for the river's name of "Big Muddy, " Most badlands silt is
now caught in main stem reservoirs.
The practice of irrigation also brings much silt into streams with
excess water returned to the rivers. It also builds up mineral content,
and frequently adds phosphorous, nitrogen, and other algal stimulants.
POLLUTIONAL EFFECTS
Pollutional effects upon aquatic life vary with nature and quantity
of waste discharge, stream stage, season, type of stream, and other
factors. Rarely does only one kind of waste originate in one locality,
and it is usually necessary to consider combined influences of various
pollutants.
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For example, oil refinery wastes generally eliminate most aquatic
animals for varying stream distances; organic wastes normally change
the character of the bottom and its biota, and eventually stimulate
plankton and other algal growth; and release of both types of waste
from one locality frequently results in alteration of the usual effects
of each In several Missouri Basin streams, waste effects are
further influenced by irrigation practices,
The North Platte River in Wyoming and Nebraska has been utilized
for irrigation since 1850. Development of the reservoir system began
in 1909 and has continued to date. Operation of this river consists of
storing runoff during seasons of greatest snow melt and later releasing
it for irrigation during dry periods. Discharges from upstream
reservoirs, frequently used for power generation, are caught in lower
reservoirs, whose releases are directly concerned with supply of irri-
gation demands,, Drawdown of lower reservoirs provides capacity for
storage of power releases in nonirrigation seasons. The operation
envisions maximum power generation consistent with necessary con-
servation of water for irrigation. With the exception of relatively
insignificant amounts of ground water inflow, discharge is wholly
controlled by reservoir releases. Reservoirs have been noted to in-
crease ground water discharge in areas just below dams. Dams without
power generators are usually cut off completely at the end of the
irrigation season; and the stream below must subsist on limited ground
water inflow:. Below such dams sudden transitions from high-to-low
or low-to-high water stages are the rule, and they involve overnight
changes from big river to headwater conditions, and vice versa.
Raw municipal sewage and oil refinery wastes enter the North Platte
at Casper, Wyoming, about 50 river miles below Alcova Reservoir. Thi
reservoir has power generation planned for the future, but at,present
operates solely to supply irrigation water, and has restricted or no
releases at other seasons. In 1950, Alcova discharges varied frorp 0 to
more than 5, 000 cfs„ During the period of high discharges, roughly May
to mid-October, biotic influences of the Casper pollutional load consisted
largely of marked plankton suppression by oil refinery discharges,
followed at some interval by stimulation from nutrients added in municipj
sewage. The plankton population developed in Alcova Reservoir, and its
concentration gradually declined in the stream until affected by waste
components. When Alcova releases ceased in October, the plankton
algae were replaced by benthic growths in shallow areas below Casper,
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These algae attained very dense development in the rich medium
(discharge was then less than 200 cfs) but were intermittently
eradicated for short periods of time by releases of phenols and crude
oils. Their photosynthesis promoted supersaturated oxygen
concentrations for many miles downstream. The oil refinery dis-
charges, with periodic releases of crude oil and caustics, had
eradicated all animal life in 114 miles of stream below Casper,
except for certain fishes that led a short-lived existence near the
mouths of some tributaries. Organic sludge deposits were completely
untenanted by sludge worms and other characteristic organisms. This
sludge and that arising from soda-lime water softening in refineries
built up concentrations during low flow periods that were scattered
downstream for about 100 miles during later flow increases.
Irrigation degradation of the river became progressively greater
with downstream distance. Its most obvious influences were accelerated
hardness, alkalinity, and turbidity increases. Silt accumulations sup-
pressed benthic organisms, and suspended sediment reduced plankton
concentration.
The sugar beet industry discharged processing wastes to the lower
North Platte during fall and early winter. Cold water at that time slowec
decay of beet particles and other organic matter, and oxygen >yas not
completely exhausted,, Flume water containing beet washings added
considerably to the river's turbidity. Irrigation ended before the beet
processing campaign began; but high river stages were maintained by
return of ground water surcharges contributed by irrigation. Except
for regions below beet factory discharges, the lower river was generally
clear in fall and winter and promoted benthic algal growth, Lime
slurry from beet processing added to alkalinity and hardness, and forme
unsightly bottom deposits inimical to benthic fauna. Many solids
contributed by the beet industry were caught in Kingsley Reservoir.
Their decomposition near the bottom of this lake provided nutrients
that stimulated algal growth in the river below. Discharges largely
consisted of bottom waters that were generally excluded from plankton
productivity in the impoundment. Storage in this reservoir reduced
hardness and alkalinity in 1950, and it discharged better quality water
than it received from upstrearru
Prairie streams lacking the benefits of mountain snow melt permit
much more limited water use than occurs along the North Platte. The
James River, arising in the prairies of central North Dakota and joining
the Missouri below Yankton, South Dakota, has been described as the
longest non-navigable river on earth.
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It is fed by prairie snow melt and limited summer rains, and
normally dries or partially dries over long reaches in early autumn.
The main flow at such seasons comes from municipal sewage and
water discharges in reaches below Huron, South Dakota. The stream
bed has a very flat gradient, and tributary inflows have been observed
to run upstream during seasons of high runoff. Several low head
dams have been constructed to conserve water for municipal usage,
stock watering, waterfowl refuges, and other purposes. The river
meanders extensively--requiring 710 channel miles to traverse a
distance of about 350 miles. Channel restrictions induce extensive
inundation of the flood plain in early spring.
The James drains an area of rich soils and has a naturally high
biological productivity. Pollution consists largely of municipal sewage
treatment plant effluents. Oily discharges from railroad yards and
artifical gas plants affect some reaches; and waterfowl have marked
effects in and below some impoundments. The usual effect of sewage
plant effluents and waterfowl wastes is excessive stimulation of plankton
algae, which frequently occasion tastes and odors in water supplies
drawn from the river. Effluent from one overloaded sewage plant
promotes anaerobiasis in the river--producing an oxygenless zone along
one side for a few miles that is followed by a region of dense plankton
growth, Tars contributed by a gas plant at that locality have eradicated
bottom organisms; and the oxygenless zone is, with the exception of
fungi, a lifeless area. Benthic organisms, especially midge larvae and
water mites, attain very dense development in areas influenced by
concentrations of waterfowl. The Sand Lake Migratory Waterfowl Refuge
northeast of Aberdeen, South Dakota, has had transient populations of
more than one-half million waterfowl, mostly geese. Due to the number
of impoundments, all waste influences are largely localized; and
plankton growth usually results in very effective treatment of wastes in
each impounded stretch.
The Yellowstone River, although supporting considerable areas of
irrigated lands, lacks impoundments. Alleviation of irrigation effects
by reimpoundment of water is, therefore, not possible. Thus, water
quality deteriorates with downstream distance in lower reaches when
irrigation is practiced.
The Yellowstone is also polluted by sugar beet factories, oil
refineries, packing houses, and treated and untreated municipal sewage.
Oils and tars originating in petroleum refineries have almost completely
eradicated bottom fauna in some reaches. Municipal wastes have been
noted to lower oxygen and stimulate algal growths.
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Periodic taste and odor problems in water supplies withdrawn below
Billings result directly and indirectly from waste discharges.
An unusual fish kill affecting long reaches of the Yellowstone was
indirectly occasioned by aerial spraying with DDT to control spruce bud-
worm in headwater forest areas. The application was followed by heavy
local rains that carried insecticide into the upper Yellowstone system.
The DDT was applied in July 1955, and a great mortality of fishes
occurred in October and,November. Autopsies of fishes indicated
starvation as the cause of death. Examination of the streams disclosed
a paucity of food organisms, mainly acquatic insects, whose widespread
scarcity was then traced back to insecticxdal operations in July.
Another unusual case of pollution resulted from practices at a
trout hatchery that permitted excess fish food and wastes to reach Rapid
Creek in western South Dakota. These materials caused deleterious
algal blooms in water supplies taken from the creek, necessitating
revision of fish rearing procedures at the hatchery. Fish protection is
normally considered an objective of water pollution control, but here
fish actively contributed to water pollution.
The above examples are offered to illustrate some pollutional
problems related to geography and human practices in the Missouri-Basin.
Space does not permit reference to various other pollution cases that
have been encountered and water quality relationships involved in the
main stem reservoir system. The heavy waste load discharged to the
lower main stem promotes taste and odor problems in water supplies
during low winter discharges, particularly if flow is reduced by ice
formation. It also maintains a high concentration of coliform-type
bacteria over the reach from Sioux City, Iowa, to below Jeffe.rsqn City,
Missouri. Effects upon the biota in the lower reach are incompletely
known. The more desirable fishes, walleyed pike, channel catfish, etc.,
are most concentrated in the river above Sioux City, while carp form
the bulk of the fish catch in polluted reaches below that point.
Water pollution in large areas of the Missouri Basin may appear
quantitatively insignificant when compared to that contributed in more
heavily industrialized regions. However, limited water supply over,
such areas results in serious effects from amounts of pollution that
would not occasion critical conditions in some areas of great rainfall.
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CURRENT INVESTIGATIONS IN WATER POLLUTION BIOLOGY
Investigations and Problems in Ohio
John N. Reis
Ohio Division of Wildlife
Delaware, Ohio
Prior to white man, it is reported that there was an approximate pop-
ulation of 10,000 Indians whose lives were, generally speaking, based on the
factors of self-preservation and actual needs. By 1800, there were some
45,000 whites who introduced the factors of trade and commerce.
By 1950, 150 years later, the human population had increased about 176
times the 1800 population to a number of almost 8 million. It is estimated
that by 1975, the population will have increased to 10 million.
Prior to the coming of the whites, practically 90 percent of the land
was covered by hardwood forests . It may be said that deforestation had begun
about the year 1788. By 1880, the entire state was settled, and some land
had already been abandoned.
Ohio has turned from an agricultural to an industrial economy. By 1959
or shortly after, the Great Lakes - St. Lawrence waterway is expected to
be open, which will have an indeterminable, but probably considerable, im-
pact on the sociological-economic status of the state..
During the early 1900's, water treatment and purification plants were
built to safeguard human health from effects of sanitary wastes. By the
1930's, the situation had progressed to the stage at which it was recognized
that action was needed to control degradation of the water courses. World
War II checked progress temporarily, after which construction of waste
treatment plants again began to pick up. In 1951, the Ohio Water Pollution
Control Act became a law giving further impetus to corrective steps being
taken.
Under this act, now on the statutes as Chapter 6111.01 to 6111.08
inclusive, a Water Pollution Control Board was established in the State De-
partment of Health with the Director of Health as chairman, the other members
being the Director of the Department of Commerce, the Director of the De-
partment of Natural Resources, a representative of industry, and a represen-
tative of municipal government. The Board administers the pollution abate-
ment program of the State.
This Board is vested with a number of duties and powers . Briefly:
1, To develop programs for prevention and control of new and existing
pollution. (Pollution means placing of any noxious of deleterious sub-
stances in any waters of the state which renders such water harmful
or inimical to the public health, or to animal or aquatic life, or to the
use of such waters for domestic water supply, or industrial or agricul-
tural purposes, or for recreation.)
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2. To advise and consult with other agencies in furtherance of the
program.
3. To encourage and conduct studies concerning pollution.
4. To collect and disseminate information concerning pollution.
5. To prohibit or abate pollution by issued orders dependent on certain
specified limitations.
6. To issue permits based on compliance with specifications.
7. To institute legal proceedings to compel compliance with the statute!
Although the Water Pollution Control Board is nominally the pollution
control agency, it is still an infant organization which is cautiously developin
its program with an eye towards continually improved effectiveness rather
than sudden blundering moves which might cause rejection of its purpose.
"Industrial wastes and acid mine drainage are exempt from the provisions o-
Section 6111. 04 of the Revised General Code of Ohio until the Water
Pollution Control Board after a hearing determines that a practical means
for the removal of the polluting properties of such wastes". "Removal" is
interpreted in Administrative Resolution No. 1, of the Water Pollution
Control Board, to mean any procedure applied to a waste which will effect
a reduction in the polluting quality 'of that waste. The program operates
under a permit system which generally does not specify the amount or
characteristics of discharges.
The Board deals with 140 cities (actually 118, since 22 are suburbs),
of which 39 do not have treatment works, but construction, planning, or
investigations are under way to correct this status.
There are 767 villages, 380 of which have been exempted as having no
pollution problems to date; .105 have been postponed because of insignificant
pollution or difficulty in financing treatment works; 22 are still to be investi
gated. Of the 114 which have been found to need installation of treatment
works, all are under construction or planning operations.
The Board deals only with industries which discharge wastes directly
into waters of the State. Of roughly 13, 000 listed industries in the State, the
Board is dealing with only 646. Of this number 283 are "currently acceptabJ
181 need improvement, and 182 are now constructing, planning, or making
preliminary studies. Those industries which discharge wastes into municip
sewers are a primary problem of the city but are also under the super-
vision of the State Health Department.
At first, the Board began bringing industry into the program by compul-
sory hearings. Following several such hearings, industrial representatives
suggested that this implied that industry was not willing to join in the abate-
ment effort, and was as a result receiving unwarranted bad publicity. Date
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lines were set and industires were allowed to voluntarily apply within that
period. All the industries asked into the program under this system are
reported to have complied.
The bulk of the Board's information concerning the polluted status
of waters is obtained from the Ohio Department of Health. This data
generally consists of B. O. D. , coliform index, acidity, tastes and odors
(phenol tests particularly), suspended and dissolved solids, temperature,
and chemical determinations for specific elements. These are generally
directed towards human health maintenance. Surveys have been made of the
Mahoning, Miami, Maumee, Muskingum and Cuyahoga Rivers.
The Division of Wildlife, Ohio Department of Natural Resources is
responsible by statute for the protection and preservation of wild animals.
Because of the preponderance of observable effects result in fish kills,
the majority of the investigations are carried out by fish managers and
game protectors.
The variability of conditions attendant to pollution incidents, of course,
necessitates adaptation of procedures to fit the circumstances. The basic
procedure is for the county wildlife management agent (known also as the
game protector) to notify the district office that a kill has occurred and then to
proceed to determine the extent and source until the fisheries technician
arrives. Together, an effort is made to pinpoint the source of pollution,
to take samples indicative of conditions, such as, pH, dissolved oxygen,
temperature, and turbidity tests, and to make counts of the observable dead
wild animals.
This report is submitted to the Division's Office of Pollution Abatement
which is also notified of the occurrence of the incident at the initiation
of the investigation. This office analyzes the information obtained in the
field and recommends the action to be taken. A claim for damages is
first presented to the offender, and if settlement cannot be reached within
30 days the claim is certified to the Ohio Attorney General for collection.
In 1955, 111 incidents were reported. Sixty-two.were reported as harmf
and forty-nine as threatening situations. A break-down of the information
concerning the sixty-two harmful incidents shows the following data in
rounded figures:
Forty percent reported counts or expressed kill figures ranging from
5 to 10, 150 fishes. The highest monetary value was reported as $36. 22
for an estimated kill of 2, 289 fish.
Thirty-one percent of the reports showed no counts, because of such
matters as too lengthy a period between occurrence and discovery of the
kill, non-feasibility of producing creditable estimates because of stream
conditions (flooding, natural swiftness, etc. ).
Nineteen percent reported damage to habitat, but as yet no basis for
monetary value has been developed for collection of these damages.
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Five percent of the kills reported were "natural" kills. White
bass, Lepibema chrysops, died in great numbers withno determinable cause
in Lake Erie. A severe water temperature change is believed to have
caused the death of numbers of smelt, Osmerus mordax. A small kill
was imported in a lake inwhich no pollutional discharges could be found.
Three percent reported that the fish had disappeared by the time of
the investigation.
Two percent reported no identifiable pollution, although a kill had been
reported.
Information obtained from an investigation by the Division is forwarded to
the Water Pollution Control Board.
Because it is obvious that present procedures are not entirely adequate)
the Division is constantly attempting to improve on its effectiveness. An
intensive study of acid mine drainage effects on Raccoon Creek was made
to determine the effect of mining operations on a stream and the possibility
of reclaiming the watershed; investigation of feasible corrective measures
are now under way. An intensive study of the effect of land use improvements
is being conducted on the Little Miami River. Statewide pollution investi-
gators whose duties are to supplement and augment the work of regular
pollution investigations and to carry out special investigations have been
added to the staff of the Office of Pollution Abatement. The Fish
Management Section in Wildlife District Two has carried out an investigation
showing the improvement on stream fish life after the installation of a
sewage treatment plant; data showed a species number increase from one
to a more normal population of 18 species in a period of three years. As
an initial program an Aquatic Biology Laboratory is being developed in order
to provide more adequate information concerning the effects of pollution
on fish life.
Other investigators and their investigations of which I am aware are:
Professor Charles Riley, Kent State University, is investigating
for the Ohio Reclamation Association, the reclamatioa of coal mined
areas by the formation of ponds in abandonned cuts and plantings on spoil
banks.
Dr. W. D. Sheets, Ohio State University, Waste Treatment
Laboratory, using a moderate sized pilot plant for the studies, is investig3-^11®
the effects of plating metals on sewage treatment installations.
Dr. George E. Barnes, Case Institute of Technology, is directing
the investigation of the effects of industrial wastes on the biota of sewage
systems in an effort to determine the effects of industrial wastes introduced
into a municipal system.
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The Ohio River Valley Water Sanitation Commission, headquartered
in Cincinnati, is an eight state organization of the Ohio River basin. Its
role is to abate and prevent pollution of this great river.
Our problems are quite similar to those concerning other states:
1. A rapidly growing population together with ever increasing indus-
trialization. The topography of Ohio is such that, except perhaps for
some of the area in the southeastern hilly region, there is little area which
can be exempted from development.
2. A need is felt for developing more effective methods of demonstra-
ting harmful effects of pollution on wildlife.
3. A revision of the status has made the interpretation of their mean-
ing and precedence of one section over another more difficult.
REFERENCES
Ohio Wildlife Resources, (Ohio Division of Wildlife, Columbus, O. :
195571 __
Clean Waters for Ohio, (Ohio Water Pollution Control Board, Columbus, O.:
1952) Vol. 1, No. I ; VoT" 2, No. 1; Vol. 3, No. 4; Vol. 4, No. 4.
Pollution Abatement Office Report of Activities, January - December, 1955,
(Ohio Division of Wildlife, Columbus, O. ) mimeographed.
Water Pollution Control Act of Ohio, (Amended Substitute Senate Bill
No 62, 195I).
Handbook of Operations, (Ohio Division of Wildlife, Columbus, O.)
not for distribution.
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BIOLOGY AND WATER POLLUTION IN GREAT BRITAIN
Thomas W. Beak
Consulting Biologist
Hawkesbury, Ontario
In a general way I think it is true to say that the trends of biological
studies in problems of water pollution in Britain have been largely influenced
iy the legislation controlling river pollution. This is not surprising as most
if the applied work in this field has been carried out by government agencies.
It means, though that a brief review of the legal aspects of pollution is
lelpful to the understanding of the biological work that has been and is being
lone in Great Britain in connection with it.
The problem of river pollution, like many other social problems, arose
vith and out of the industrial revolution. By the middle of the nineteenth
century it had become serious in the industrialized central belts of England
tnd southern Scotland. A series of Royal Commissions were set up to study
tnd report on the situation and the last of these remained in being until
he early 1920's. These Royal Commissions carried out a great deal of
applied research on river pollution and much of the basic work in this field
ncluding I believe the development of the five day B, O. D. test - was done
'y them. As a result of these studies they suggested standards for effluent
'Urity depending on available dilution, but their recommendations were not
arried through to legislation to any great extent.
In 1876, before most of the Royal Commissions' studies had been made,
River Pollution Prevention Act was passed, but it was really an adminis-
rative rather than a technical Act. In effect it made any sort of pollution illegal,
ut, as the complete prevention of pollution and the existance of industry and
'ban living were incompatible, the Act provided legal loopholes that were
o large that the whole Act proved almost useless as a means of preventing
dilution.
This remained the legal situation until 1951 when two River Pollution
Prevention Acts, one for Scotland and one for England, became law. These
>cts were very similar technically, the differences being chiefly in the bodies
barged with the administration of the Acts. For the first time in Great
'*itain these Acts accepted the principle of allowing a controlled amount
f pollution and authorized the setting up of standards of purity to which
fluents must comply. As these standards necessarily vary from one river
3 another and even between different parts of the same river, provision was
Jade for them to be enacted by means of by-laws of limited local operation
My.
Apart from the early work carried out by the Royal Commissioners, the
^nistry of Agriculture and Fisheries employed a team consisting of Southgate,
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Pentelow and Butcher to carry out stream surveys in the 1920's and 1930's.
These surveys , as one would expect from the later history of the men employed
on them, were very thorough and provided a valuable basis for biological
survey work in connection with river pollution.
One of the recommendations of the Royal Commission on Sewage Disposal
that came to fruition was the setting up of a permanent research laboratory
to work on water pollution problems, namely the Water Pollution Research
Laboratory (9) of which Dr. Southgate has been Director for many years.
Particularly since the second world war the study of pollution problems
in Great Britain has very largely centered on this laboratory and in collab-
oration with the Fisheries Inspectorate of the Ministry of Agriculture and
Fisheries, of which Pentelow is Chief Inspector, a considerable amount of
research is at present in progress .
The programes of research have been shaped very largely to meet the
requirements of the 1951 legislation already referred to, and particularly
to enable standards of purity to be established. The approach to this
problem has been four-fold. First, one team which includes Herbert, Merkins ,
and Kathleen Downing has been carrying out fundamental researches on
the toxicity to fish of various chemicals, in particular cyanide, ammonia
and carbon dioxide (2) (4) (5) (6) (7) (8). For this work a very fine piece
of apparatus for controlled flow experiments has been developed and used.
Another team including Herbert, Alabaster and Allan has been working
on the field aspects of the problem by studying rivers in various degrees
of pollution and also by conducting field experiments with fish kept under
pollutional conditions^.). Alabasters, in collaboration with other workers has also
been engaged in the development of a standard method for measurement of
toxicity and although I am not aware that he has yet published his results,
he has accumulated a considerable amount of data.
Apart from these purely biological researches, some of the chemists and
physical chemists - particularly Downing, Gameson, Knowles and Truesdale -
have been working on the problems of re-oxygenation, de-oxygenation, oxygen
sag curve and other physical and chemical problems that are very closely
related to the biological aspects of river pollution^).
There has been a great deal of discussion at this seminar on the pro-
blem of bioassays and toxicity tests . It may be of interest for me to give,
as far as I know it, the British approach to the problem(5).
First there seems to me to be a difference in the goals of the workers
in Britain and the United States of America. In Britain it has been assumed
that toxicity tests will be carried out by a specialist agency, so that the
aim has been to develop a practical and reproduceable test that can be
handled by a laboratory specially equipped and staffed for the purpose. In
the United States the aim seems to be to develop a test that can be carried
out by an industrial control laboratory that is probably not specially equipped
or staffed for the purpose .
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The British workers have not favoured the method which has come to
be known as the Doudoroff, Katz method of retaining fish in solutions of
poison through which air is bubbled to maintain oxygen concentrations and
measuring 50% tolerance limits, because they felt that the shortcomings of
the method, which were of course known and stated by its originators, were
as difficult to overcome as the problems posed by some of the more compli-
cated experimental procedures. The shortcomings to which I refer are, of
course, the removal of volatile materials by aeration and the difficulty of
prescribing safe limits from 50% tolerance limits. Instead the approach
of the British workers has been, first by means of the fundamental work
of Herbert, Merkins and Downing and the field work already mentioned to
try to learn sufficient of the mechanism of toxicity to enable safe limits
for toxicity to be calculated from comparatively short term tests, probably
measuring median survival times rather than 50% tolerance limits. Secondly,
attempts have been made to develop an experimental technique that will
enable these measurements to be made without aeration. Controlled flow
seems to be the most practical line of enquiry and I believe that current
work is taking place along these lines.
Apart from the work being carried out by the Water Pollution Research
Laboratory in conjunction with the Ministry of Agriculture and Fisheries,
there are other workers in Great Britain interested in freshwater biology
who may from time to time carry out work having a bearing on pollution
problems but, as far as I know, none of them specialize in this work. The
Freshwater Biological Association of the British Empire, which has its
headquarters on Lake Windemere does a great deal of valuable research
of a rather more fundamental character than the Water Pollution Research
Laboratory. The Scottish Home Department has a Brown Trout Research
Laboratory at Pitlochry, Perthshire, but it is concerned more with fishery
improvement than pollution. Some of the universities have freshwater
biology stations; for example: Aberystwyth, where Kathleen Carpenter
carried out much of her early pioneer work on freshwater ecology, Liverpool,
Glasgow and Wessex - formerly University College Southampton - to mention
a few that happen to be known to me personally.
I should like to make it clear that this statement does not purport to
be a comprehensive review of the biological work in connection with river
Pollution in Great Britain. It is merely the observations of one person
who has been connected with this work for many years and has tried, often
'ainly I fear, to keep in touch with the work that is being done in this field. I
•rust that no one will be hurt by my omissions but will attribute theit> to tiny
ignorance of their work, not to any disparagement of it.
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REFERENCES:
1. Allan, I. R. H., Alabaster, J. S. and Herbert, D. W. M., (1954) Recent
studies on toxicity and stream pollution. The Water and Sanitary
Engineer 5(3), 109.
2. Downing, K. M., (1954) The influence of dissolved oxygen concentrations on
the toxicity of potassium cyanide to rainbow trout. Journ. Exp. Biol. 31,1®*'
3. Gameson, A. L. H,t Truesdale, G. A. and Downing, A. L., (1955) Re-
aeration studies in a lakeland beck. Journ. Inst. Water Eng. 9, 571 .
4. Herbert, D. W. M. and Merkens , J. C .,(1952) The toxicity of potassium
cyanide to trout. Journ. Exp. Biol. 29, 632.
5. Herbert, D. W. M., (1952) Measurement of the toxicity of substances to fisb'
Jour, and Proc. Inst. Sewage Purification (1), 60.
6. Herbert, D. W. M., Downing, K. M. and Merkens, J. C., (1955) Studies on
the survival of fish from poisonous solutions. Proc. Intern. Assoc.
Theor. and App. Limn. 12, 789-
7. Herbert, D. W. M. and Downing, K. M., (1955) A further study of the
toxicity of potassium cyanide to rainbow trout. Ann. Appl. Biol 43 (2),
237.
8. Herbert D. W. M., (1955) Measuring the toxicity of effluents to fish.
The Analyst-80 (957), 896.
9. Southgate, B. A., (1955) The water pollution research laboratory.
Laboratory Practice 4 (6), 257.
10. Southgate, B. A. and Gameson, A. L. H., (1956) Recent developments in
the control of stream pollution. The Surveyor 115 (3344), 349.
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THE TRAINING OF AQUATIC SANITARY BIOLOGISTS
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THE TRAINING OF AQUATIC SANITARY BIOLOGISTS
Curtis L. Newcombe
U.S. Naval Radiological Defense Laboratory
San Francisco 24, California
INTRODUCTION
Our subject for discussion this evening is certainly not the least in
mportance among the six panel discussions that comprise this Seminar
>n Biological Problems in Water Pollution. The success of the training
irograms of today will be reflected in the solutions of tomorrow's problems.
As new problems arise in science and industry, new or revised training
>rograms are born. There is nothing unespected about that, rather it is a
latural educational trend. There was a time, not too long ago, when a sanitary
sngineer as a separate species, did not exist. Today, many of our larger
iniversities have separate departments of Sanitary Engineering.
But when it comes to sanitary biology the problem is not so simple.
--et us consider for a moment two somewhat analagous training problems
rising from the expansion of two fields of study—first, Conservation and
econd, Operation Analysis or what the British call Operational Research.
Dwindling national resources accentuated the need for conservation,
idvocates of one school of thought favored treating conservation as a more
r less discrete entity to be presented or taught as a separate subject or
eries of subjects. A second viewpoint advocated the teaching of conserva-
Lon as an integral part of practically all other courses of instruction. After
luch debate, this latter viewpoint has received widest acceptance.
On a basis of the diversity of subject matter presented during the past
aur days it is evident that aquatic sanitary biology is indeed a broad field,
equiring basic training in mathematics, chemistry, and physics as well as
number of branches of biology.
I propose, as a basis for discussion by this panel, that a sanitary biolo-
y curriculum be composed of strong basic courses in each of the afore-
lentioned subjects followed by one or possibly two courses designed par-
cularly for the sanitary biologist. Incorporated in these latter courses should
e the history of advances in this field, the biological contributions responsible
>r research accomplishments to date, the status of today's research in this
eld and the future trends indicated by current investigations. Also, these
ourses should set forth the problem needs of the engineer and attempt to
ridge any existing gap between these respective disciplines.
What we might term a second reference field is Operation Analysis,
is of course a very much newer field than conservation — an outgrowth
military research experience during World War II.
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Operations Analysis may be defined as a discipline for planning actions
most effectively for some purpose. Optimization of achievement of purpose
is stressed. It may involve a problem in reducing traffic casualties or diag-
nosing the source of pollution in a river. In every case, it involves an action
taken for a purpose and involving expenditure of effort ¦
We may decide, let us say, to allocate a certain amount of effort to
accomplish a mission. First, we must know the real specific purpose of
the operation. We must establish a problem model. The term model is
not used here in the sense of a small scale structure but rather as a thinking^
device with which to analyse a situation. It is important to devise effective
models in approaching a pollution problem or any other kind. Such models
are, of course, approximations. Their upper limits or permissible boundaries
must be specified.
Thus, from operations research, we learn to construct a problem
model, to analyse it, to find out how sensitive our conclusions are to this
model.
Presently certain colleges are grouping courses to form special curri-
cula for training students in Operations Analysis .
One highly significant outgrowth of experience to date in Operation
Analysis is that investigators representing unrelated disciplines have attacked
problems and solved them although these problems have been entirely new
and most unique. This is attributed to a fresh and unprejudiced viewpoint
provided by the unindoctrinated researcher. To preserve this "take nothing
for granted atmosphere", curriculum designers have an understandable hesi'
tancy about prescribing special courses in Operation Analysis.
We are here this evening to consider problems similar to those faced
by curriculum makers in the fields of conservation and operation research.
Among other things:
(1) Shall we train aquatic sanitary biologists by introducing special
courses ?
(2) by modifying existing course?
or (3) shall we simply regroup unmodified existing courses ? and come
with a cross fertilization that may meet the need.
If we aim to give our undergraduate major and masters degree work
as a prerequisite to taking a position in the field, what training courses and
fields should be included?
What additions should be made for students continuing for the doctorate?
Are our students to be trained strictly as applied researchers or
we make the program flexible?
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What provision can be made for enabling the student to get one or two
semesters of actual field experience in sanitary biology before completing
his training?
BASIC CONSIDERATIONS
Having dealt with a number of general considerations and alternative
approaches to the problem of adequate training of aquatic sanitary biologists,
we need now to formulate specifically our problem. Let us consider the
objectives of the training program. It has been said that the Case Institute
is successful because it turns out what industry wants.
What does the sanitary engineer want from a sanitary biologist? I
would like, Mr. Chairman to hear this question discussed.
I'm sure he would, among other things want a biologist capable of recog-
nizing the central problem, of evaluating the techniques most likely to be
effective in solving it, and then capable of applying those techniques.
The biologist must recognize alternative courses of action, he must
recognize different measures of effectiveness, let us say, in respect to al-
ternative control measures, he must be able to analyse and evaluate the
variables involved, he must be capable of careful theoretical analysis and
nnost important be able to subject his data to adequate statistical analysis.
We should perhaps at this point expell the notion that our task here is
unique. The correct answer, if there is one, to this training problem may be
unique but the problem itself certainly does not stand alone.
In common with chemists, physiologists and teachers of other sciences,
we aim to graduate scientists first. Our student product must have those
earmarks of a trained mind, one capable of clear and logical thinking for
which there is no substitute in the evolution of mankind.
Ernst A. Hauser of M.I. T. states^ it well "The true purpose of educa-
tion should not be to make living textbooks, so to speak, rather we must
change our curricula so that more emphasis is placed on the disciplines that
teach proficiency in doing and thinking."
I believe therefore that the objective in our training program should be
to train students to think and do as scientists first and then as sanitary
biologists.
Engineers often use public moneys. They need to know what represents
^e best bet. They will ask what are the best probabilities in selecting an
alternative.
I have heard it said that during the war a statistician undertook to evaluate
'low effective British bombing missions were in helping Russia. Number of
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casualties was the objective. They started by determining the effect in England
of German raids. It was found that 5000 tons of bombs were dropped on
England yielding casualties of 0.8 persons per ton. Due to various factors
the Germaji equivalent was estimated to be 0.4 persons per ton and later this
figure was reduced to 0 .2. In other words 400 civilians were killed per
month. Then it was realized that, since 40 British bombers each with 5
trained men were lost per month, it therefore required a loss of 200 trained
personnel to produce 200 civilian casualties. Not a very encouraging result.
Further after the war it was learned that they had been using a wrong measure
of effectiveness. Spread-out bombing operations would have been more ef-
fective than concentrating on particular centers as was usually done.
This story illustrates, I think, the importance of defining one's problem,
evaluating the effectiveness of alternative methods of action or control.
In grouping of training courses to comprise a sanitary biology curriculut**'
I would attach primary emphasis on the basic sciences prerequisite to aquatic
sanitary biology—physics, chemistry, bacteriology, biometry and the calculus
accompanied by a number of biology courses which I have elected to leave
for discussion by others on this panel better able to present this part of the
subject.
Allow me to venture the opinion that too many of our institutions have
gone overboard in the offering of specialized courses. Not that I have any-
thing against the refinement of departmental offerings. They, of course,
have a place. That definitely is not to serve as a substitute for courses
that are more effective in terms of the principal purpose of the program in
question. In other words first courses should come first. Thinking capacity'
has priority, over textbook mastery.
Again, I would like to hear our panel discuss this thesis: Are we
training our students adequately to solve problems? I say emphatically, no.
We complain that our current graduates of high schools are steering away
from science. If this is true, there doubtless are a number of causative fac-
tors. I think the problem approach could well be stressed. Further I believe
curriculum makers could well mold their offerings in such a way as to get
our graduates to recognize and solve problems. Ability to use the tools
of basic science, to focus them on a practical problem, such as how to de-
sign a more effective or efficient oxidation lagoon, requires more than rout**1
classroom work in a variety of academic courses. First it presupposes BOt&e
experience in the practices of thinking or reasoning. A problem such as the
lagoon one mentioned, here, is no respecter of courses. Varying discipli*1®
are represented.
In fact, we are in this panel, faced with a problem in operation research-
We must define our training problem—our training objective, or purpose,
examine alternative means of meeting the objective, undertake to express
or state the anticipated effectiveness of the one or more alternatives.
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Advances in sanitary biology will depend upon having the best brains
and upon having a large corps of dedicated investigators to enlarge the
advances. Paraphrasing the words of Paul Weiss 12) of the University of
Chicago: "All it takes to join these forces is: some aspirations, to point
the goal; some inspiration, to point the way; and a lot of perspiration, to
get the job done."
REFERENCES CITED
(1) Hauser, Ernst A. 1951. The Importance of Science in American
Education,Science, 113, 2945, p. 646
(2) Weiss, Paul 1953, The Challenge of Biology,Science, 118, 3054, p. 34.
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THE JOHNS HOPKINS UNIVERSITY
SCHOOL OF ENGINEERING
Department of Sanitary Engineering
^nd Water Resources
Baltimore 18, Maryland
509 Ames Hall
April 23, 1956
Dr. Clarence M. Tarzwell
Robert A. Taft Sanitary Engineering Center
4676 Columbia Parkway
Cincinnati 26, Ohio
Dear Tarzie:
I don't see how I can get to the Thursday seminars and back here to
keep appointments that have piled up.
I have a few comments on the training of aquatic sanitary biologists
that represent my own prejudices in the matter. They grow from limited
local experience and from my total ignorance of the requirements and
operations of the official agencies.
1. What are our future requirements for such a specialist group?
It would appear that the demand for sanitary biologists specializing
in aquatic biology would be determined by employment in state and
federal agencies and by a more limited use in industries that have
continuing stream pollution problems. I would hazard the guess that
federal demands would not exceed 50 men and that the state demands
under most active conditions would be about the same level. It is
possible that half this many men would be employed by industry in one
capacity or another --a total of 150 men. This can be quite wrong, of
course, and the number might be doubled or halved. Some unexpected
legislation might, on the other hand, raise the number to a total of
1, 500 or 3, 000.
In any case, it would appear that aquatic sanitary biology represents
a small profession, far less numerous than sanitary engineering or
water works operation. The turnover in this total profession determines
the need for special training facilities. If we assume the largest likely
number, 3,000, and take an annual turnover of 10%, the yearly require-
ment would be around 300 to be divided up among the various training
facilities in this country. The annual requirement, then, is quite small
and well within the scope of a comparatively simple special training unit.
If the turnover is larger, say 25%, we have demands of another order,
and also evidence that the profession is a fairly unsatisfactory one.
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Dr. Tarzwell
1.
April 23, 1956
2. Why do we need special training?
Special training seems to be required to permit useful integration
of biological observations with other criteria of water quality and
changes used by sanitary engineers. The sanitary engineering profession
has definite ideas of what biologists are and how they should function
in a program. The role of biologist is, however, definitely accessory.
The general history of stream pollution investigations in this country
has shown a strong movement away from time consuming biological
evaluations to the apparently more quantitative evaluations given by
sanitary chemistry. In most sanitary engineers' thinking, the biologist's
job is to find suitable indicators for water quality. If these correlate
well with B. O. D. and other characteristics of the water, he is happy.
If they do not, he doesn't know what to do about it and is inclined to
drop the matter.
With increase in new industrial wastes, a second major use of
biologists has developed -- testing for toxicity and tolerance. Dramatic
toxicity results are, of course, big medicine but the ecological
evaluation of waste loadings is, as every field biologist knows, much
more complicated.
In any case, the special training that is required of biologists appears
to be the development of an understanding of sanitary engineering
terminology and of the peculiar whims and policies of the "team*1. This
may take some time. Undoubtedly, the best way to establish this
rapport is through direct training in the field unit. Classroom train-
ing tends to remove itself from current professional uses. Most of
our programs that would employ sanitary aquatic biologists grow from
special appropriation projects. These are organized from the top
down— the demands on the biologists will be determined by the condition*
of the appropriation. Heaven knows what these may be, but they need
not be wise or far sighted.
3. What is the role of universities in the training of sanitary
aquatic biologists?
The general history of biology departments has shown movement
away from natural history toward physiological, biochemical, and
genetics type departments. There are few universities with strong
natural history divisions. While the natural history courses are by
nature intriguing, they have not had the professional potential of other
branches -- generally speaking, the professional potential of biology
majors has been very low. A few years ago more than half of our
"professional biologists" were employed in the Department of Agricult-
ure. These are perhaps the most happily used men in our university gr°
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Dr. Tarzwell
3.
April 23, 1956
Industrial uses have been extremely limited and biological courses
have not attracted men who expected to enter industry. This restriction
is becoming more apparent and the stature of biology generally has
suffered.
The value of any special training offered in a university depends
first upon the quality of the men who are attraced to it, and secondly,
upon the quality of the men who do the teaching and their understanding
of the total problem. It may not be flattering to say that we do not get
the most brilliant and promising students in public health, conservation,
aquatic biology and other courses that might be conditional to sanitary
aquatic biology specialities. It is more realistic to admit that the
brilliant students simply don't appear in these courses. Our reservoir
of talent is severely limited.
With the decline of natural history training, biology departments
have lost contact with the field. Although many members of departments
serve on meritorious committees and pass large policy statements,
comparatively few work directly with either official agencies or with
industries. They are not acquainted with the values involved in any
realistic way.
4. How can the training of aquatic sanitary biologists be improved?
It would appear that there is little justification for setting up special
courses in universities for training biologists to work in official agencies
where the law enforcement component dominates. A better use of
limited talent is in the prosecution of special investigations and examinat-
ion of new phenomena. Here the most resourceful people are demanded.
My own feeling is that we can do most by making it possible for biology
department members to see what the problems in sanitary biology are
really like; to study them in their own way, and to reevaluate them in
an original fashion. The official point of view is secondary here. In
the development of this relationship industry should take a very active
part, especially in those states where conflict of recreational and in-
dustrial values is likely. The constitution of a special sanitary biologi-
cal profession perpetuates the concept of public health losses (as a
level to forward conservation interests) and it is doubtful if these men
will be happy in the promulgation of this type of compromise. It is very
valuable to universities to have men active in industries and biology
departments would gain stature considerably by such contact. We have
had numerous instances of men initially involved in pollution studies
who were able to make real contributions to the solution of production
problems in industry. This transfer of ability increases the respect of
industry for biologists.
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Dr. Tarzwell
4.
April 23, 1956
5. What students should be encouraged to enter fields related
to aquatic sanitary biology?
It is fairly obvious that men who are to be happy and successful
in this field must be capable of growth. Service in a bureau at levels
currently reserved for biologists is far from satisfactory. This
grows in large part from the limited functions of bureaus. Generally
speaking, official agencies lose their most enterprising men to in-
dustries and to private organizations because these men prefer the
freedom that is possible in private work. At the same time, apprentice-
ship in bureau service provides an essential background for successful
private practice in companies that must maintain contact with official
agencies. It is unlikely that the future will differ from the present,
although salaries and fringe benefits may make federal and state service
more attractive. Two possibilities exist. A man may serve as a highly
specialized expert or he may be required to exercise a variety of
techniques. The specialist, of course, is subject to limitations of
shifting emphasis. A taxonomic entomologist may be in great demand
in early phases of an insect borne study, and find himself on the shelf
a few years later when practice has been established. It takes along
time to train a good taxonomist. The more generally trained man enters
a more varied field of experience. He is usually better adapted to
"the team" operation, although personality problems always enter.
It would be most unhappy if sanitary aquatic biology became a
catchall for misfit biologists. This could be possible and I am afraid
that the possibility would be increased by organizing the field in a
series of academic offerings, especially if good beginning salaries and
security were offered as additional inducements for entering the field.
The problem of finding professional outlets for biologists is not peculiar
to the public health and sanitary fields. We must make an effort to
analyze the whole employment situation to determine what the best
uses and futures may be. Very few biology departments have practical
contacts of an order that permit good student placement and student
advising.
With all good wishes,
Very sincerely,
sgd/ Charlie
Charles E. Renn
Professor of Sanitary
Engineering
CER:mh
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WATER QUALITY CRITERIA FOR AQUATIC LIFE
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EFFECTS OF TURBIDITY AND SILT ON AQUATIC LIFE
John N. Wilson
U. S. Public Health Service
Portland, Oregon
Mr. Chairman and Gentlemen of the Seminar, I'm happy to
have the opportunity to participate in this panel on water quality
criteria and to present to you some of my experiences and those of
others dealing with the effects of turbidity and silt on aquatic life.
Upon my arrival in the Pacific Northwest as biologist with the Drain-
age Basin Office, U. S. Public Health Service, several years ago,
one of my first tasks was to work with the committee on Water Quality
Criteria and Objectives for the Pacific Northwest Pollution Control
Council, At that time we sidestepped the question of effects of
turbidity and silt on aquatic life. Under the heading of "Floating,
Suspended, and Settleable Solids and Sludge Deposits" we stated: "None
shall be permissable which are attributable to sewage, industrial
wastes or other wastes or which after reasonable dilution and mixture
with receiving waters interfere with the best use of these waters for the
purpose indicated. 11
A review of water quality criteria and standards from all over the
country discloses a reluctance, if not outright refusal on the part of
regulatory agencies to come to grips with this problem. Many consider
the control of silt and turbidity to be strictly a matter for agriculture
departments -- indeed for the farmer himself -- and are therefore un-
willing to touch it as a pollution control problem.
A matter of semantics is involved at this point as many persons
such as those connected with the industries of mining and forest products
adhere to the strictest connotation of the word "pollution. " They consider
silt and turbidity as perfectly natural and normal to be carried by water
in contrast to pollution which is contended by them to refer only to
putrescrible matter and denoting transportation by water of pathogenic
bacteria and the like. However, the currently accepted definition as
reiterated by most forwardlooking pollution control agencies refers to
pollution of water as encompassing the full gamut of substances which
adversely affect any of the legitimate uses of the water.
In view of the fact that there is widely differing opinion throughout
the country, and the world, in regard to how deleterious silt and
turbidity are in natural waters, a committee has recently been organized
with the author as chairman to work on a "Critical Review of Represent-
ative Literature on Water Pollution by Inert Inorganic Materials. " This
committee includes Dr. A. F. Bartsch of the Robert A. Taft Sanitary
Engineering Center; Dr. I.E. Wallen of the American Association for
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the Advancement ol Science, wasnmgton, u. , ui . vugu j-ion.,
University of Idaho, Moscow; and Mr. Philip Pister of the California
Fish and Game Department, Garberville, California. We have not
made remarkable progress as yet but have set three years as our
goal, hoping to have reasonably full coverage by that time, leading
toward publication.
My presentation to you should, by rights, be made three years
from now when our committee has had an opportunity to complete its
work. Nevertheless, we can all profit at this time by a brief discussion
of some findings to date. I shall refer to some pertinent activities and
publications in the field, including an investigation which I have made
with the State of Oregon on effects of gold dredging on an eastern Oregon
stream.
Jacob Verduin (1954) at Stone Lab, Put-in-Bay, Ohio, states that
turbidity is the major factor in causing poor phytoplankton productivity
in Lake Erie.
David Starr Jordan (1889) reported many decades ago on losses of
trout and trout spawn owing to turbidity and silt from placer mining
operations in Colorado.
Richard Rathbun (1889) in writing of streams in Iowa, tells of change
with development of agriculture causing decrease and deterioration of
the better food fishes. With the breakup of original sod of the prairies,
rivers which formerly had well-defined, deep, narrow channels have
widened and become shallow. They tend to overflow their banks in the
rainy season and lose most of their water during succeeding months.
Sediment and silt, continually loosened by farming, fill pools and
riffles in streams, thereby causing rapid disappearance of trout. We
are presumably to consider the trout which formerly occupied streams
in Iowa as passe just as are the buffalo from the western plains.
Higgins (1931) referred to the enormous quantities of erosion silt
entering the Mississippi River along with sewage sludge. Together
these agents smothered bottom life, wiped out mussel beds, and the
like.
Emmeline Moore (1937) quotes H. K. Townes in saying that silta-
tion dilutes settling organic debris to such an extent that bottom
organisms have a poor food supply. There is a continual "snowing
under" of bottom organisms. In fact, induced siltation has been used
in irrigation canals to inhibit growth of moss and algae.
Tarzwell, (1937) in the course of stream improvement work in
Michigan, evaluated the effectiveness of the structures and came up
with some enlightening facts on productivity of stream bottom types.
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At the lower extreme of the scale is sand with population rating of
1 and silt 10.5; rubble, 30; coarse gravel, 32; but plant beds provide
the very highest productivity with ranges up to 452. A normally
productive riffle with coarse gravel and rubble will depreciate in
productivity stages toward 1 as the riffle fills with sand and silt.
Langlois (1941) pointed out changes in fish fauna in lakes and in
streams with ecological changes in these habitats. In lakes, reductior
in species of fish is seen, while in streams there may be a replaceme.
of some fish species by others. Those fish that can reproduce under
the greatest variety of conditions persist the longest; those requiring
the most specific conditions are the first eliminated. Attempts to re-
store fish species that have been decimated or eliminated should
consist of restoration of the habitat conditions that prevailed when tho«
species were thriving.
Bartsch and Schilpp (1953) reported on sand processing wastes fr
a glass sand corporation in West Virginia as affecting a small tributar
of the Potomac River , They concluded that the differences in product:
of plants and animals in affected and unaffected parts of the river are
due principally to increased turbidity and solids deposition.
They further concluded that "the physical and biological conditions
found to occur in affected parts of the river are ones known to interfer
with sport fishing and to affect adversely the production of fish. "
The color slides which I have presented show some of the trouble
spots in the Pacific Northwest in this matter of siltation and turbidity.
Particular reference is made to the situation on the Powder River in
eastern Oregon where a gold dredge had been operating for a number
of years.
By good fortune I was able, with the help of Messrs. Homer Camj
bell of the Oregon State Game Department and Harold M. Patterson of
the Oregon State Sanitary Authority to investigate the Powder River in
September and October 1953 before the dredge ceased operation and
again in November 1955, more than two years after the dredge stoppe<
Our summary and conclusions at the close of the first phase of the stu
were:
1. Turbidity readings ranged from 5 ppm to the control area to
1700 ppm below the dredge. Siltation in pools was very heavj
2. A test for rate of sedimentation on a sample of water from be-
low the dredge indicated a 60 per cent reduction in suspended
solids in 24 hours.
3. Production of fish-food organisms dropped to almost nil in the
zone of heaviest pollution.
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4. The presence of some bottom fauna at all stations indicates
absence of toxic substances in the mining waste. Principal
damages are physical smothering action and interference with
light penetration necessary for growth of green plants.
5. Results of fish population studies in the various zones of
pollution in Powder River indicated a complete alteration of
the population from sport fish, rainbow trout and whitefish,found
above all sources of pollution to rough fish in the zone of
pollution and recovery.
The sedimentation test which was made on waters of the Powder
River downstream from the gold dredge indicated a high degree of
reduction in suspended matter in a 24-hour period. During stages of
normal to low flow a corresponding progressive decrease in suspended
matter was observed downstream from the source of pollution. Although
the screening of the light was a significant factor in lowered biological
productivity, the abrasive or molar action of the larger particles of
sediment and the smothering of fish food organisms and fish spawning
beds are considered to be of greater importance.
The more recent investigation showed that remarkable recovery
had taken place in the Powder River with flushing of silt from the pools
and cleansing of riffles by freshets. This signalled the return of a wide
variety of bottom fauna in the 15-20 mile reach of the river which had
been heavily silted. The Oregon Game Commission has planted trout in
this stretch and creel census indicates successful reestablishment of the
sport fishery.
This brings us back to our original thesis, namely: The establishme
of water quality criteria for silt and turbidity in natural waters. May
I be permitted to incorporate here the consensus of opinions expressed
by members of the seminar audience? It has been suggested that rather
than to propose arbitrary criteria either for turbidity or for settleable
solids, some percentage increase above normal low flow concentrations
should be established. This would take into consideration differences
in watershed and stream or reservoir characteristics.
In conclusion, the problem of siltation and excessive turbidity
stemming from activities of man are widespread and difficult to control-
Many question the economic feasibility and practicability of control
measures, but a few forward-looking pollution control agencies are
undertaking measures of control with varying degrees of success. As a
guiding principle in establishment of water quality criteria for permis-
sible concentrations of silt and turbidity in streams, certain percentage
increases above levels at normal low flow in waters is suggested.
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BIBLIOGRAPHY
Bartsch, A. F., and J. H. Schilpp 1953. Water pollution survey of
the Potomac River Hancock-William sport Section. A coopera-
tive report by State of Maryland, Water Pollution Control
Commission; West Virginia, State Water Commission and
North Atlantic Drainage Basins Office, Division of Water
Pollution Control, Public Health Service.
Higgins, Elmer 1931. Progress in biological inquiries, 1930. U. S.
Bureau of Fisheries, Ann. Rept. 3:553-626, illus.
Jordan, D. Starr 1889. Report of explorations in Colorado and Utah
during the summer of 1889, with an account of the fishes
found in each of the river basins examined. U.S. Fish Comm.
Bull. 9:1-40, illus .
Langlois, T. H. 1941. Two processes operating for the reduction in
abundance or elimination of fish species from certain types of
-water areas. Trans. Sixth North Am. Wildlife Conf. 189-201.
Moore, Emmeline 1937. The effect of silting on the productivity of
waters . Trans . Second North Am . Wildlife Conf. 658-661.
Rathbun, Richard 1889. Report upon the inquiry respecting food fishes
and the fishing-grounds. U.S. Comm. of Fish and Fisheries,
Ann. Rept. 17:97-171.
Tarzwell, C. M. 1937. Experimental evidence as to the value of trout
stream improvement in Michigan. Trans. Am. Fish. Soc., 66:
177-187.
Verduin, Jacob 1954. Phytoplankton and turbidity in western Lake Erie.
Ecology 35(4):550-56l.
Wilson, J. N. 1953. Report on biological reconnaissance on the effect
of gold dredging and mining operations on Powder and Burnt
Rivers, Oregon. A cooperative report by the Oregon State
Game Commission, Oregon State Sanitary Authority and
Division of Water Pollution Control, Public Health Service.
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THE EFFECT OF POLLUTION UPON WILDLIFE
O. Lloyd Meehean
U.S. Fish and Wildlife Service
Washington, D. C.
I am pleased to have this opportunity to join you to learn of the many
problems facing those working in the field of pollution and to learn of the
progress being made in the technical field toward the alleviation of these
problems . It goes without saying that the advance of civilization has brought
with it many wonders, but at the same time has left in its wake many things,
one of the worst being pollution, which are in contradiction to the many
fine recorded accomplishments .
As my portion of this discussion, I shall attempt to summarize for
you the effect of pollution upon aquatic life as it relates to our objectives
and functions in the field of fish and wildlife conservation.
The programs of the Fish and Wildlife Service have the central objec-
tive of insuring the conservation of the Nation's wild birds, mammals,
fishes, and other forms of wildlife both for their recreational and economic
values . In carrying out this objective, which we share with the States and
Territories, it is essential to build up these resources, to prevent their
destruction or depletion, and to promote the maximum use and enjoyment
in consonance with their perpetuation.
Before discussing the Fish and Wildlife Service's interest in the field
of pollution as it relates to wildlife, I shall review briefly the functions of
the Service, particularly those which are concerned in one way or another
with pollution problems .
The conservation of the North American waterfowl and other migratory
birds is undertaken in cooperation with the governments of Canada and
Mexico. The Service operates 264 refuges to provide for the needs of water-
fowl and other species as a major part of this program. Our investigations
and surveys are the principal basis of the hunting regulations, which are
enforced by the Service.
The Service is charged with the administration and enforcement of the
laws relating to the commercial fisheries of Alaska. It also conducts
biological research on marine species of commercial importance off all
coasts of the United States and in waters adjacent to territorial possessions.
The fishery research program provides information on the size of the re-
sources rates of decline or increase, and reaction to various intensities
of fishing as a basis of conservation programs. The marine fishery program
also includes technological, economic, and statistical research as means
of promoting trade and commerce in fishery products, improving processing,
distribution, and marketing practices, and effecting more complete utili-
zation of resources.
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Our programs for the maintenance of the fresh-water fisheries extend
to the conservation and development of commercial fisheries of the Great
Lakes and other inland waters, and to cooperation with the States in the
maintenance of sport fishing throughout the Nation. The maintenance pro-
grams for Federal lands and installations depend upon stocking from the
Service's hatcheries and upon our programs for the protection and restora-
tion of habitat. Much attention is given to investigation of the effects of water
development projects proposed for construction by the Federal Government
or under Federal permit or license. In a number of instances the Service
maintains and operates fish screens on Federal irrigation projects .
The Service administers the Federal Aid in Fish and Wildlife Restora-
tion Acts, which authorize grants-in-aid to the States and Territories, in-
cluding investigations, acquisition of land, and development of fish and
wildlife habitat. The Service is authorized to provide assistance to, and
cooperate with, Federal, State, and public or private agencies and organi-
zations in the development, protection, rearing, and stocking of all species
of wildlife and fish.
Among other important programs of the Service are activities relating
to international agreements concerning fish and wildlife, including the Whaling
Treaty Act, the Sockeye Salmon Fishery Act, and the North Pacific Hali-
but Fisheries Act; the management of the North Pacific fur seal herd; the
administration and enforcement of the various Federal laws relating to
wildlife and fisheries, including the restraints upon interstate transporta-
tion contained in the Lacey Act, and the Black Bass Act; and the promotion
of domestically produced fishery products in commerce, the development
of markets for fishery products of domestic origin, and the conduct of
research pertaining to American fisheries.
There is seemingly no limit to the varieties of pollution which direc-
tly or indirectly are harmful to fish and wildlife resources. Despite this,
there is no Federal authority under which the Service enforces regulations
governing pollution.
Since we lack the power of enforcement, we must, when the need arises,
seek the aid of State and Federal agencies who are empowered with such
authority. Our activities in the pollution field are confined largely to re-
search on special problems and to cooperation with State and other Federal;
agencies . Without the excellent cooperation received from these agencies
in working out solutions to our problems, there is no question that our
progress in fish and wildlife conservation would be seriously retarded.
To keep abreast with pollution problems, the Service is represented on
interagency committees which consider water-development planning. We
are also represented on a National Research Council Committee which stud*el
and reports on the effects of radioactive wastes on aquatic life, and on the
Interdepartmental Committee on Pest Control.
In our investigations of water-development projects proposed for con-
struction by the Federal Government or by private interests under Federal
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permit or license, we attempt to recognize existing pollution problems and
consider the possible influence which the project will have on existing pollu-
tion conditions and the consequent effects upon fish and wildlife. After con-
sulting the States, the Public Health Service, and others, recommendations
aimed at avoiding future detrimental effects on these resources are made.
In many cases it is difficult to identify a water-development project as a
direct cause of pollution, although the operation of these projects would
rlearly worsen the existing problem. The Roanoke River in North Carolina
ran be cited as an outstanding example of the need for industry and State
ind Federal Governments to join forces in conquering a severe pollution
aroblem created by wastes from paper mills and municipal sewage, and aggra-
vated by regulation of flow and by the low oxygen content of water released
it the Roanoke Rapids Dam. Among other effects, the striped bass fishery
a threatened. Here, the forces have joined together to make studies of the
situation, and we are hopeful that the many competing uses for water can
ae satisfied through an abatement program and modification of reservoir
operations .
While on the subject of water-development projects, I should like to point
>ut that some projects threaten fish and wildlife resources in certain coastal
ireas . By changing flow regimen or by re-routing waters, the usefulness
>f certain marsh areas for waterfowl, other wildlife, and fish may be des-
royed as a result of saltwater intrusion into fresh-water areas. In other
nstances, estuarine environments may become less saline as a result of
ncreased flows of fresh water and seriously affect fish and shellfish pro-
iuction.
A type of pollution that has become one of our most pressing problems
s that resulting from the increased use of insecticides, herbicides, and
ungicides . It is estimated that farmers, home gardeners, and government
>rganizations charged with control of noxious insects and plants now pur-
:hase 700 million pounds of these basic materials each year. This represents
ibout 3 billion pounds of finished pesticides that are sprayed or dusted
mnually on millions of acres of the Nation's crop, forest, range, and marsh
ands.
Aquatic organisms, particularly fish, crabs, and insects, are among
he most sensitive of all animal life to poisoning by pesticides. Insecticides,
;specially the chlorinated hydrocarbon group, offer the greatest hazard,
ierbicides are generally less toxic, but in addition to direct poisoning they
lestroy both aquatic and terrestrial food and cover plants. Other groups of
>esticides, such as fungicides and rodenticides, present much less danger
mder conditions of use.
Our wildlife research on the biological effects of pesticides has bee*
:onfined principally to the effects of insecticides on wildlife species inhabit-
ng marsh areas. Considerable of our attention in these studies has necessaril
ieen devoted to determining effects of these materials on lower forms of
ife because of the importance of many invertebrates as food for birds and
nammals .
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For example, fiddler crabs comprise about 90 percent of the summer
diet of clapper rails in New Jersey salt marshes . Cooperative investiga-
tions conducted with mosquito control agencies in Atlantic coastal marshes
have shown that single applications of 0.4 to 0 .5 of a pound of DDT in one-
half to one gallon of No. 2 fuel oil per acre produced moderate to severe
but temporary losses of fish, crabs, and other invertebrates, while applica-
tions of 0.2 to 0 .3 of a pound of DDT in one-half to one gallon of No. 2 fuel
oil per acre repeated several times a summer for several years caused
significant, more lasting damage to most of these forms. There was little
direct harm to birds and mammals although long-term routine spraying
operations appeared to reduce carrying capacity for these higher animals.
The studies also revealed that DDT in granules caused less damage than
equivalent dosages in oil solution. Of the newer insecticides, dieldrin ,
aldrin, and lindane were more toxic than DDT to crabs.
Experimental feeding of these chemicals, including DDT and strobane,
to quail and pheasants decreased the number of eggs produced and reduced
the hatchability and viability of the eggs. The viability of the young was also
reduced. These effects were observed when concentrations in the diet
were as low as 1 ppm of aldrin, 5 ppm of dieldrin, 50 ppm of strobane, and
100 ppm of DDT. It has been reported that application of 1 pound of these
chemicals per acre under agricultural practices would result in concentratior
of 50 ppm in seeds and vegetation which may be eaten by wildlife.
The Service has recently been requested to participate in an investiga-
tion of fish kills in the Upper Yellowstone River area which possibly re-
sulted from DDT spraying operations for the control of the spruce budworm.
In 1955, a high mortality of whitefish and a lesser mortality of trout and
suckers occurred in the stream several months after spraying operations
in July. Destruction of fish food organisms resulting in fish starvation may
be a contributing factor. Although similar spraying in previous years did
no apparent damage, this serious incident signals the need for studies which
will give us a better understanding of DDT formulations and their effects
on fish and wildlife .
Pollution from industrial sources includes a wide variety of substances
which are lethal or harmful to fish and wildlife. Among the serious offenders
are the wastes from paper and pulp mills, sawmills, chemical manufacturing
plants, textile plants, mines, oil fields and processing plants, sugar beet
processing plants, meat packing plants, dairy product processing plants,
leather processing plants, and many other operations.
The Service has no continuing research program to study the lethal
or undesirable characteristics of these wastes upon fish and wildlife, so
we look principally to the research studies of others for this type of infor-
mation. Some of this information is sought by the States under the Federal
Aid in Fish Restoration programs which the Service administers. Most
of these studies, however, are more in the nature of general pollution
surveys.
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From about 1941 to 1946, the Service conducted surveys of some of the
sources of pollution of this nature and made studies in an effort to determii
minimum quantities which fish could withstand. I believe most of you are
aware of the difficulties in setting up standards which would apply to a
variety of conditions .
This spring, studies by the Service are being initiated in Kentucky in
cooperation with the U.S. Public Health Service, the U.S. Geological Surve
ind the State of Kentucky to determine, under controlled conditions, the
affects of coal strip-mining pollution on stream ecology. Since this pollutit
has a definite limiting effect on the productivity of the aquatic environment;
these studies will serve as a basis for developing recommendations to
protect or improve fish and wildlife habitat which may be, or is, affected
ay such operations.
There are numerous reports of the deleterious effects of mine wastes
apon fish and wildlife. Annually, thousands of waterfowl seeking aquatic
:ood and fish fall victim to poisoning. Such a situation occurs regularly
in the Coeur d'Alene River in Idaho. The ore deposits in the area are pri-
marily sulfides , chiefly of lead and zinc associated with deposits of silver,
:admium, bismuth, arsenic, antimony, and iron. Sulfides of various heavy
•netals become even more dangerous when they change over to other com-
Dounds . Little progress has been made during the past 30 years in getting
:he pollutors to remedy these hazards. In addition, mine slimes have creat
less productive aquatic environment in the river and in Coeur d'Alene Lake
The pollution situation was studied and reported on by the Service in 1932
ind we encourage a study which was inaugurated a year ago by the Coopera
;ive Wildlife Research Unit at the University of Idaho.
Oddly enough, even the hunter accounts for a significant amount of
iead poisoning in waterfowl. Birds feeding in water areas which have been
leavily hunted, swallow enough shotgun pellets to result in quick death.
Oil pollution not only destroys our marine and inland fisheries, but
results in a heavy mortality of waterfowl. In many cases the oil pollution
is from the discharge of barges, ships, and accidental or careless handling
jf oil transportation, development, and drilling operations . Waterfowl aligl
tng on oil sump areas or water areas covered with oil are usually rendered
'lightless. Oil is also a destroyer of aquatic life including foods of both
"ish and wildlife. Even in minute quantities oil may impart undesirable
:astes to the flesh of fish and shellfish, thus rendering them unpalatable as
luman food. In cases which come to our attention, the Federal and State
authorities are contacted. The Corps of Engineers and Coast Guard are oft
:ontacted in such cases relating to navigation under the Oil Pollution Act
Df 1924, which is administered by the Department of the Army. Oil operati
Dn certain of our wildlife refuges are kept under close surveillance to avoi<
damage to waterfowl.
On many areas where serious pollution hazards of various types occur
it is necessary for our game management agents to keep the waterfowl driv
"rom these areas.
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Silt and washings from mining operations are 01 mucn concern iu ub.
Dredging and reworking of stream bottoms for gold and other minerals
brings indiscriminate destruction to fishery resources and habitat. The
turning over of the stream bottoms destroys important spawning and food
producing areas, and silt from such operations quickly snuffs out the lives
of incubating fish eggs and aquatic organisms . Salmon and trout on the
West Coast and in Alaska come in for their share of punishment from
these operations. It seems that solutions to some of these problems could
best be solved through the withdrawal of public lands along streams to ex-
clude detrimental mining operation. Such a procedure has been attempted,
but has met with considerable opposition from mining interests.
In Alaska, fishery resources are not protected by the Alaska Pollution
Act in the case of silt from mining operations . The Act specifically states
that "the results of activities connected with gravel washing plants and all
phases of Placer Mining Operations shall not be considered pollution within
the meaning of this Act."
Service hatchery and planting operations are sometimes handicapped
by silt from lumbering and other operations. Certain situations occurring
on U. S. Forest lands have been remedied by reporting such matters to the
Forest Service which took steps to assure that the lumber operator complie<
with the terms of his contract.
Logging and pulp operations have long presented serious problems to
fish and wildlife. This industry is relatively new in Alaska, where the effect
are now being felt by the salmon fishing industry. A study will be conducted
by the Fisheries Research Institute of the University of Washington in coope
tion with the Service to determine the effects of these operations.
Municipal pollution, with particular reference to sewage, brings great
concern from the standpoint of health, but it also may seriously affect fish
and wildlife. In some instances small amounts of organic pollution may be
beneficial to fish, but high concentrations may deplete the oxygen supply
and result in fish kills or the loss of food organisms . The Service relies
heavily upon the States and the Public Health Service to bring about the
abatement or elimination of pollution from such sources. There have
been occurrences, however, in which the Service's installations such as
refuges have been affected, and the Service has encouraged cities to install
treatment plants.
As you can readily see, the Service does encounter many pollution pro-
blems affecting its interests, few of which can be resolved without assistant
of other agencies. The men who come up with the answers to the technical
problems deserve a lot of credit in the total effort aimed at beating pollution
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WATER QUALITY CRITERIA FOR AQUATIC LIFE
Clarence M. Tarzwell
Chief of Aquatic Biology
Robert A. Taft Sanitary Engineering Center
Public Health Service
Cincinnati, Ohio
The lakes and rivers of North America have played a very important
role in the opening and development of the continent. In our Great Lakes we
have a fresh water resource far surpassing any other in the world. Our rivers
and hundreds of thousands of inland lakes are outstanding in their many uses
and support a diversity of the most valuable fishes to be found anywhere. Per-
haps it is because of this great wealth in aquatic resources, which many
considered to be inexhaustible, that we have been so remiss in their protection
and conservation. Their importance is only now coming to be generally
appreciated. The great increases in population since 1900 and the manifold
Increases in the volume and variety of water uses have shown that in some
areas the supply of water is definitely limited. As with other resources we
find that value varies directly with demand and inversely with supply. In the
western areas which recently suffered from a severe drought it was found that
when local drinking water supplies dry up,water will be purchased at any
lecessary price, however high it may be.
What is true for drinking water is also true in some measure for
iquatic life resources. We have already found that as the demand for de-
sirable fishing increases and the supply diminishes, the amount paid for such
recreation becomes greater. The development of the country has drastically
reduced or eliminated fishing waters in extensive areas. Deforestation, fire,
ivergrazing, and unwise agricultural practices have increased surface runoff
ind decreased seepage causing floods, intermittent flow, the drying up of
springs, erosion, silting, and the filling of stream beds. Removal of stream-
side vegetation has promoted bank erosion, the widening of streams, and
varming of the water. Industrial and other pollution has blocked fish migratior
ind has rendered many areas unsuitable for fish. These practices which
liter or destroy the aquatic habitat are the chief cause of the decline of aquatic
ife resources. The surest way to eliminate a species or group of species
s to destroy their habitat or produce environmental conditions unfavorable
or them. The only way to maintain a species is to protect and maintain
snvironmental conditions essential for and conducive to its growth, reproductio
md well being.
Protection and conservation of aquatic life is not a simple task,
iome have questioned its value and feasibility. Our fisheries are now an
mportant resource. The commercial fisheries of the United States and
Uaska have an annual production of almost five billion pounds (1). This
:rop represents a high protein resource which can be expected to increase in
ralue. While commercial fisheries utilize a great deal of equipment and emplo
arge numbers of people, their economic worth is only a fraction of that of spoi
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fishing. In 1955 there were more than 17 million licensed fisherman in
the United States. There are several million additional fisherman who fish
in the Great Lakes and in marine waters where a license is not required.
According to the National Survey of Fishing and Hunting (2), sport fishing
in the United States has an annual value of almost two billion dollars. Within
the next 100 years the number of fishermen and the value of the fishery will
increase severalfold if the resources can be preserved.
In comparing the beneficial uses of a stream, there has often been
a tendency to underestimate the wildlife and recreational values and to take
a short rather than a long view. When evaluating our fisheries resources we
should consider the returns not just for one year but over the centuries be-
cause these resources are renewable. The aesthetic, recreational, and
health values of our waters are difficult to measure, but they are great. The
recreational industry is a large one and is expanding every year. In a few
states it is the first ranking industry and in many others it is of considerable
importance. In the industrial state of Michigan it is reputed to rank second.
As our population increases there will be an ever growing demand for and use
of our forests, parks, preserves, wilderness areas, and streams, where peopl*
can engage in water sports and "get back" to nature and relax. It is believed
that our aquatic life resources and the aesthetic value of our lakes and streams*
which are largely inseparable, are well worth our earnest and sincere efforts
to preserve them.
The objective of water quality criteria for the protection of aquatic
life is to preserve or restore environmental conditions essential for its
growth, reproduction, and well being. If these requirements are known and
understood, criteria can be set up which will achieve this objective. If
habitat requirements are not fully known, criteria can only be based on the
best information available and changed whenever the need is indicated by
new information.
Under our present state of knowledge a suitable water cannot be
defined in chemical terms alone. There are several reasons for this situation-
Different species of fishes and the organisms in their food chain vary widely
in their sensitivity to dissolved materials. We do not know the effects on
aquatic life of various concentrations of many materials individually or in
combination. Mixtures of materials often have effects different from those
of the individual components. Further, we do not know minimal lethal levels
for many materials or their mixtures, nor do we know the most favorable
concentrations of materials essential for the organisms. Perhaps the best
definition that can be given of a suitable fish habitat is -- "A suitable fish
habitat is one which produces a satisfactory fish crop. " The adequacy of a
fish crop is judged by its quality and the pounds produced per unit of surface
area. Commonly, productivity and suitability are judged by catch per unit
effort, growth rate, condition factor, quality of the flesh, and the size and
species composition of the catch. In fisheries management an effort is
made to manipulate the environment so that conditions are made more favor®**1
for the desired species and less favorable for those not wanted.
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While our knowledge of the habitat requirements of fishes is far from
complete, enough information is now available so that some criteria and
procedures can be set up which will be of value in the maintenance of a satis-
factory environment and production of a suitable crop. However, much research
is still needed to obtain all the information essential for the solution of this
problem.
The environmental requiremenjsof fishes may be roughly grouped under
four main headings, (1) a favorable water supply; (2) suitable spawning
facilities; (3) an adequate food supply for all age groups; and (4) good pools
and shelter. In the abatement and control of pollution we are chiefly concerned
with the first requirement, a favorable water supply. Natural waters have
widely varying physical and chemical properties. The suitability of any water
for fish life depends on its quantity, permanency, and quality; that is, its
temperature, the concentration of dissolved atmospheric gases, salts and other
minerals, and suspended solids.
As here used water pollution is the addition of any material or waste
to a water in such quantities that it interferes with, lessens, or destroys a
beneficial use. In this regard perhaps the simplest definition of pollution is
"too much. " For example, if "too much" is not added, the discharge to a stream
of organic matter such as sewage has a fertilizing effect which is beneficial
to fish production. However, when the capacity of the stream to utilize
organic materials is exceeded so that unfavorable environmental conditions are
produced and a beneficial use is damaged, such a discharge becomes pollution.
It is evident, therefore, that water quality criteria for the protection of aquatic
life must entail some quantitative measurements. Before criteria can be set
up we must know or have some measure of how much is "too much" for those
species we wish to protect at all stages in their life history and in waters of
different quality. These criteria must insure environmental conditions favorable
to all life activities and to general well being—mere survival is not enough.
These environmental factors will be discussed in some detail.
Environmental Factors
Temperature
A s a group, aquatic animals of the temperate zone are adapted to
fluctuations in temperature between 39° and 90°F, Not all can withstand this
range and some can withstand higher and some lower temperatures for a time.
The range of temperature which can be tolerated by different species varies
considerably as does their ability to withstand sudden changes or to acclimate
themselves to unusual temperatures. Each species has a preferred range of
temperature within which it does best and a zone above and below this in which
it can survive for short periods. Proper acclimatization enables certain species
to survive at temperatures which would be fatal under conditions of sudden
exposure.
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Fish have a rapid rate of adaptation to high temperatures (3) but
adaptation to lower temperatures proceeds at a much slower rate. When
suddenly exposed to higher temperatures fish can withstand much higher
temperatures in summer than in winter. As the summer season develops,,
changes in upper lethal temperatures reflect the major changes in water
temperature, rising with ascending temperatures and falling as the water cools
in the fall. Brett (3) reports that the lethal temperatures for the bullhead
rose from 29. 1°C. on May 12 to 35. 3°C, by July 8. Brett (4) also found
that there is a considerable difference in the time required for completion of
acclimation with respect to heat tolerance at each level of temperature. It
has been shown that some species exhibit geographic differences in their
resistance to high temperatures while others do not (5). The writer noted a
bass kill in a southern Michigan lake in June 1936 when the water reached 94°®*
during an unusually hot period. However, a temperature of 96°F. in Wheeler
Reservoir in 1938 appeared to have no lethal effects. The difference may
have been due to a different acclimation history. It was observed that a
temperature of 108°F. killed all the fish in a pond near Savannah, Georgia, in
1945. Allowable peak temperatures brought about by some unnatural cause
may, therefore, be somewhat different in different portions of the country,
increasing from north to south.
Members of the family Salmonidae are cold water forms. Brook trout
seem to do best in streams, the summer temperatures of which range between
52 and 68°F. (6) (7). While trout can survive much higher temperatures for
short periods, streams having such temperatures are not first class trout
streams. The writer found brown and rainbow trout surviving a peak temperati
of 83°F. in the South Branch of the Pere Marquette River of Michigan in 1930.
Brook trout survived peak temperatures of 81° and 82°F. in the East Branch
of the Black River on successive days in July 1931. Fry (8) reported the
upper lethal temperature for young brook trout (12 to 14 hours exposure) to be
about 77. 5°F. Such high temperatures are more favorable for minnows and
suckers which increase greatly in numbers and compete for food and space
with the trout (9). The result is that trout comprise a very small portion of
the total fish population of the stream and supply little fishing even though
the overall productivity of the stream may be great.
Four fish population studies made in the East Branch of the Black
River of Michigan indicated that trout made upon only 9. 6 percent of the total
number of fish taken in the study areas. In the neighboring Pigeon River,
another stream having temperatures above 75°F. , trout comprised 15 per-
cent of the total fish population in the areas counted (9). In a nearby cold
stream, the West Branch of the Sturgeon River, trout represented 96 per-
cent of the total population. In trout streams having high peak summer
temperatures, suckers and minnows comprise the bulk of the fish population-
Thus, while temperatures higher than the optimum and high temperatures oi
short duration (75® to 82°F.) may not kill trout, they produce environmental
conditions more favorable for the coarse fishes, which increase at the ex-
pense of the trout population. This fact must be taken into consideration in
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the establishment of temperature criteria for the cold water species. Such
criteria must be based on optimum conditions and not on temperatures tolerated
by trout. It is believed that for good trout production in streams subject to
invasion by coarse species, temperatures should not exceed 68°F.
Since deforestation, overgrazing, unwise land use, removal of stream-
side shade, and erosion have already caused the warming of streams to such an
extent that the amount of trout water has been seriously reduced, and since
trout fishing is in highest demand, it is believed that no wastes of significant
heat content should be discharged into a trout stream if the stream is to be
maintained for trout.
Favorable temperatures are especially important at spawning time for
both cold and warm water species. It is well known that bass spawn in the
spring when the water temperature exceeds 60°F. If the water is unnaturally
warmed to this temperature for a period, spawning may be induced too early
in the season. Then if waste discharges are discontinued over a weekend,
water temperatures may drop into the 50's with the result that guarding males
leave the nests, the eggs are infested with fungi, and no young are produced.
Fluctuations of water temperature above and below 60°F. during the spring
are detrimental to bass production.
A change in water temperature may affect the aquatic fauna directly
or indirectly. While the change may be within the thermal tolerance of the
fish it may so alter environmental conditions that they become unfavorable
for essential food organisms and certain life history stages of the fish, or
the change may make them more favorable for competitors or predators
Temperature changes will directly affect the metabolic rate, growth, and
reproductive processes. They may result in increased or decreased food
production, interfere with spawning, or change an important part of the fauna,
thereby altering the quantitative makeup of the population.
Although high summer temperatures have been considered of out-
standing importance because of their possible lethal effects, it is believed
that unnaturally high winter temperatures may be equally important. In
the temperate zone the aquatic biota have evolved under conditions of quite
wide differences in seasonal temperatures. For example, the eggs of some
daphnia have to be chilled or frozen before they hatch. Many other organisms
go through resting stages or specific stages of development at certain seasons.
Some of the diatoms, for example, are abundant only at temperatures below
50°F. Other forms appear only at certain times of the year and there is a
succession of forms with the seasonal changes. At present we have little con-
ception of the changes which might be brought about by permanently elevating
stream temperatures. A large portion of the biota might be changed and the
whole food chain disrupted. For this reason consideration should be given
to upper temperature limits during the winter season. This consideration
may require increasing attention as the atomic energy industry develops.
Temperature should not be raised to levels that induce spawning at unnatural
times if there are periodic drops in temperature, and they should not be such
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that they interfere with the development ol important organisms in me ubu
food chain. Considerable study is needed before this problem can be
approached intelligently.
Waters of significant heat content should not be discharged into a
stream in such a manner that they create a temperature block across the
stream. Further, an abrupt change of more than 9°F. may affect fishes
adversely even if of short duration.
For a well rounded warm water fish population in the Ohio valley
area it is believed that peak summer temperatures should not exceed 93°F.
at any time or place. In the south such peak temperatures probably should not
be above 96°F. This means that in general temperatures will be considerably
below these levels. While several species can withstand higher temperatures
(100° to 103°F. ) for very short periods, 93° and 96°F. represent critical levels
for most species in the designated areas. Further, while fish may, through
certain adaptations, survive abnormally high temperatures for short periods,
they cannot complete their life history at such temperatures. For good product!
therefore, temperatures within a favorable range are required.
Settleable Solids and Turbidity
Studies carried out in connection with trout stream improvement in-
vestigations in Michigan indicated that sand bottoms are almost barren of
benthic organisms and that the addition of sand or silt to rubble or gravel
bottom streams greatly decreases stream productivity (9) (10) (11). In fact,
shifting sand in quantities so small as to be unnoticed by casual observation
can decrease the production of macro-invertebrates by drifting into the spaces
between the gravel and thereby decreasing the areas for attachment and cover.
It is believed that no inert inorganic, sandy, or other similar wastes should be
added to a rubble, gravel bottom stream as such deposition may not only
decrease the supply of desirable stream bottom insects but also seriously limi*
spawning c£ most nest-building fishes. Studies of the effects of mining wastes
in California have shown that salmon select clear water for spawning and that
the deposition of silt results in smothering of the eggs (12). Quantitative
bottom samples taken in a series of similar streams in California showed that
the average number of food organisms was always less in mined areas when
inert inorganic materials were discharged to the stream than in nonmined
areas (13). On the Scott River, samples from the silted area averaged 36
organisms per square foot, whereas those from the clean stream bottom above
the mine averaged 249 per square foot, or 7 times as many. Similar studies
(14) have shown that hydraulic mining wastes are detrimental to salmon and
trout production. These and wastes from placer mining and from stamp mill®
and washing operations may completely choke a stream causing it to flow in a
shallow sheet over the accumulated deposits. Further, debris dams created
by such operations can eliminate the salmon by blocking migration.
From results of studies in various parts of the country, it is
apparent that erosion silt is a major stream pollutant and that it produces
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Environmental conditions unfavorable for the reproduction and growth of fishes.
Since the character of the stream and its bottom are of prime importance in
determining the harmful effects of erosion silt, it is not possible to establish
numerical criteria for settleable solids which are universally applicable. In
some streams considerable amounts do very little additional harm, while in
gravel rubble bottom streams even small amounts, as has been noted, reduce
food production and limit spawning. It is believed that criteria on settleable
solids should be established to protect environmental conditions in the
stream, though they will vary from stream to stream, depending on local
conditions.
Turbidity is usually due to solids which settle out slowly or to
colloidal materials which may remain in suspension over long periods. The
studies of Irwin (15), Wallen (15A), and others at Oklahoma A and M have
3hown that turbidity must be very high before it exerts a directly harmful
sffect on fishes. In some tests (lSj)direct reactions to turbidity did not
ippear until it reached 20, 000 p. p. m. and for one species not until it reached
LOO, 000 p. p.m. Most individuals of all species endured exposure to more than
L00, 000 p. p.m. for a week or longer but finally died at turbidities of 175,000
:o 225, 000 p. p.m. Fishes which succumbed to these turbidities had the
jpercular cavities and gill filaments clogged with silty clay particles.
In Oklahoma, Buck (16) carried on pond studies to determine the
jffects of turbidity on growth rate. At the end of two growing seasons the
iverage total weight of fish in clear ponds was about 1. 7 times that of those
n ponds of intermediate turbidities and approximately 5. 5 times greater
han those in muddy ponds. Of the three species used, large mouth bass were
nost affected by turbidity. The effect on plankton production was even more
striking since the average volume of net plankton in clear ponds during the
954 growing season was 8 times greater than in ponds having intermediate
urbidity and 12.8 times greater than the yield in most turbid ponds. How-
iver, catfish survived better in muddy ponds. Game fish feed by sight and
n turbid waters they are at a disasvantage when competing with such fish
is carp, buffalo, and carp suckers which employ a vacuum cleaner type of
eeding. Turbidity can, therefore, bring about a quantitative and qualitative
:hange in the fish fauna. In addition, metallic or sharp particles may kill
ishes by causing abrasive injuries to the gills or by clogging the gills and
•espiratory passages.
Suspended solids and turbidity prevent light penetration, decrease
Photosynthesis, and thus limit algal production. Since algae are the basic
naterial in the food pyramid, turbidity adversely affects fish production in
,n indirect manner. In most streams settleable solids and turbidity are
argely due to soil erosion. Until erosion is brought undsr control, little
:an be done toward clearing up the streams. Reduction of turbidity is a
lifficult and long time problem which must be carried out in cooperation with
oil conservation, agricultural, and forestry interests. In the meantime,
towever, efforts should be made to control or eliminate other sources of
ettleable solids and turbidity. Lagooning can be effectively used to remove
ettleable solids and turbidity from many wastes. Such procedures are
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essential on all clear streams and they should be initiated in conjunction
with efforts to reduce turbidity and settleable solids through control of soil
erosion.
Turbidity standards must be somewhat local in their application
as they will depend on the area and type of stream. It is possible to set up
relatively simple turbidity standards which can be readily checked for
compliance by field tests. Turbidity standards might state that a certain
percentage of the incident light at the surface shall reach a stated depthbetweei
11:00 A. M. and 1:00 P. M. The depth selected would depend on the depth
to which the regulatory agency felt the photosynthetic zone should extend.
Different types of water differ in their capacities to absorb light. Water
transparency is affected by the suspended matter, including the plankton, and
by stain or color. In water of the clarity of usual municipal supplies, 9. 5
percent of the solar energy present at the surface reaches a depth of 6 feet.
Born (39) states that the limit for growth for the higher aquatic plants lies
between 2. 5 and 3. 5 percent of the total surface energy at bottom depth, but
that it rapidly declines below 4 percent where severe etiolation occurs in
submerged seed plants. There is some evidence that certain algae can grow
at levels of 1 percent of the incident light, but it is not definitely known how
much light is required for them to produce more oxygen by photosynthesis
than they use in their respiration. While criteria will vary with the area they
can be kept relatively simple. For example, a criterion for a particular
area might state -- under conditions of brilliant sunlight at or near noon 4
percent of surface incident light shall reach a depth of 6 feet. Incident light
and light at any given depth can be readily read by means of a photometer
fitted for underwater use.
EiL
The pH of a water may exert a direct effect on fish if it is very high
or very low due to strong bases or mineral acids. It may have an indirect
effect through its influence on the toxicity of certain materials such as HCN,
H2S, ammonia, heavy metals, etc. Longwell and Pentelow (17) found that
the toxicity of NaS solutions to brown trout was influenced markedly by
variations in pH, the toxicity increasing as the pH became lower. The heavy
metals are considerably more toxic at lower pH levels probably because they
are more soluble. Ammonia becomes rapidly more toxic as the pH is raised
above 8. 2. The toxicity of a number of weak inorganic and organic acids,
including hydrocyanic, hypochlorous, hydrosulfuric, carbonic, and tannic,
is increased by lowering the pH.
Extreme pH values of 4 and 10 or slightly above have been tolerated
by resistant fishes in certain areas. Some levels at which fish have been
killed experimentally are: trout, 9. 2; bluegills, 10. 5; roach, pike, carp,
and tench, 10. 4 to 10. 8. Fish mortality has been observed within a few
hours at pH levels of 3. 4 to 4. However, certain fish have been acclimated
to live for considerable periods at pH levels as low as 4. 5 to 4. 2.
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Studies of acid bog lakes (18) have shown that yellow perch,
brown bullhead, bluegill, and pike can live at a pH of 4.4. Ellis (19) states
that the pH of streams generally ranges between 7. 4 and 8. 5 with an over-
all range of 6. 6 to 9. 0, while bog streams and lakes vary from 4. 0 to 6. 0.
He states that in most uncontaminated freshwater streams pH values range from
6, 5 to 8. 5.
Sudden or wide fluctuations in pH are undesirable. While fish
can withstand pH levels as high as 9. 5, it is undesirable to have the pH
maintained continually between 9 and 9. 5 when this level is due to the addition
of caustic wastes. Such pH conditions are entirely different from and more
harmful than the naturally occurring but brief higher levels which may be as
high as 10 or 10. 5. These natural high pH levels are produced by photo-
synthesis due to the removal of CO2 and they are always accompanied by
high D. O. levels. High pH interferes with oxygen uptake of some marine and
freshwater fishes and may limit their ability to survive at low oxygen tension
(20). At values below 5 and above 9, the pH seriously affects the ability of
some fishes to extract oxygen from the water. This ability varies with the
species; with bass and crappie the pH can be lowered to almost 4 before their
ability is affected (21) (22). In general, fish are able to extract oxygen best
at pH levels from 7. 0 to 8. 5, but such fish as perch, bass, crappie, gold-
fish, trout, and green sunfish have a wide range of tolerance. The blunt -
nose minnow and one of the shiners, Notropis whipplii, were found to be very
sensitive as they can extract oxygen best at pH 7. 0 to 8. 0 (20). Some fishes
can survive rapid changes in pH. Laboratory studies (21) have indicated
that goldfish withstood changes from 7. 2 to 9. 6, black bass from 6. 6 to 9. 3,
and sunfish from 7. 2 to 9. 6. The amount of dissolved oxygen is a determining
factor as to whether or not these changes can be tolerated (21).
In the range from 5 to 9.5, pH as such has not been shown to be
detrimental to fishes. However, changes in this range can drastically affect
the toxicity of certain materials, and they also influence the ability of fish
to absorb oxygen from the water. Further, it has been noted that in the more
productive streams pH usually falls in the range from 6. 5 to 8. 5. At pH
levels above and below these values some of the essential minerals become
unavailable. Thus, while pH in the range from 5. 0 to 9. 5 is not in itself
directly harmful to fishes and this range may be used in setting up water
quality criteria, from the standpoint of productivity, it is recommended that
every effort be made to keep pH values in the range of 6. 5 to 8. 5.
Dissolved Oxygen
There are a host of environmental and other conditions which influence
or determine the solubility of oxygen in water, the amount of dissolved oxygen
favorable to fish life, and the minimum amount needed for existence. In
fresh waters, temperature is the most important factor affecting the solubility
of oxygen. Dissolved solids are rarely present in sufficient amounts to have
an appreciable influence. Several environmental conditions may influence the
optimum amount of oxygen required by fish or interfere with the obtaining
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of oxygen by the fish or may change or increase their minimum need for
oxygen. Among these are temperature, pH, CO2, and dissolved solids.
Temperature increases within the range favorable to fish are
accompanied by a progressively higher metabolic rate and a continuous
increase in the oxygen uptake. Wiebe and Fuller (23) found that at 25°C.
the oxygen consumption of largemouth black bass was 282 percent of that
at 15°C. At 20°C. it was 177 percent of the consumption at 15°C. This is in
accord with the van't Hoff law which states that for any chemical change the
rate of reaction is increased between 2- and 3-fold for every 10 C. increase
in temperature. Temperature is of outstanding importance in the determination
of environmental requirements since the oxygen consumption increases as
temperature rises whereas solubility of oxygen decreases. Because the annual
range in temperature of streams of the temperate region may be as much as
28°C. , oxygen consumption at peak temperatures may be several fold what
it is at minimum temperatures, whereas at peak stream or lake temperatures
the water will hold only about half as much oxygen as it does at minimum
temperatures. Graham (24) found that for speckled trout the rate of oxygen
uptake increased with increasing temperature up to the ultimate upper
lethal temperature, if sufficient oxygen were available. Water containing
less than 75 percent of the air saturation level of oxygen reduced the activity
of speckled trout at all temperatures, and above 20°C. (68°F. ) fully saturated
water is required to allow the full scope of activities. Several other in-
vestigators have also found that the oxygen requirements of fishes become
greater with increases in temperature (25) (26) (27).
Temperature also markedly affects dissolved oxygen concentrations
with are lethal to various species of fish. Burdick (28) found that small-
mouth bass died in 5 to 9 hours at oxygen concentrations of 0.7 p. p* m. to
1. 17 p. p.m. at temperatures of 52°F. to 72°F. There was also some
variation in the turnover time for different species of fishes. At 55°F. and
oxygen concentrations of 1 to 2 p.p. m. the turnover times were as follows:
brook trout, 1-3/4 hours; brown trout, 2-1/2 hours; and rainbow trout, 3
hours. At 69°F. to 71 F. these fishes turned over in approximately the
same time at oxygen concentrations of 2. 3 to 3. 4 p. p. m.
Several other environmental factors also interfere with oxygen upt»ke
or increase the oxygen requirements of fishes. High and low pH levels
interfere with the ability of fishes to absorb oxygen from the water. High C°2
concentrations interfere with the utilization of dissolved oxygen. Fry and
Black (29) found that the common sucker, with its C02 sensitive blood, was
unable to remove oxygen from water containing C02 tension which did not
hinder the respiration of bullheads, the latter possessing blood with a very
low sensitivity to CO,. Under pollutional conditions fish generally require
more oxygen (45) (46J (20). At low dissolved oxygen levels fish succumb to
concentrations of toxic materials which they can tolerate at high dissolved
oxygen levels.
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Many studies have been made in attempts to determine the lowest
D, O. levels tolerated by different species of fish. Gutsell (30) reported
that some brook trout could endure, for a short period, an oxygen concentration
as low as 1. 2 p. p.m. ; however, some asphyxiation occurred at a D, O. content
of 2. 5 p. p. m. Smallmouth black bass lived for a time at 0. 4 p. p. m. D. O.
Wiebe (22) found that some fish can withstand sudden wide changes in the
concentration of oxygen and that they can live in water supersaturated with
oxygen. The increase in D. O. was followed by a slowing down of the respiratory
movements. Fry (31) states that at 49 F. the ultimate minimal tolerance of
brook trout for dissolved oxygen is 0. 9 p. p. m. Gardner and King (32) re-
ported the asphyxial level of trout to be 1.1 p. p. m. D. O. at 6. 5°C. and 3. 4
p.p. m. at 25°C. Thompson (33) on the basis of field studies, reported that
carp and buffalo lived in water having 2. 2 p. p. m. D. O. However, he found
a variety of fishes only when there was over 4 p.p. m. of oxygen and the greatest
variety of fishes were present when the D. O. was 9 p.p. m. He found that fish
died overnight in water containing less than 2 p. p.m. D. O. Ellis reported (19)
that goldfish, perch, catfish, and other species of freshwater fishes when
maintained in water of constant flow, composition, and temperature (20° to
25°C. ) showed respiratory compensation in both volume and rate when the
dissolved oxygen was reduced to slightly below 5 p. p. m.
In addition to those environmental conditions which influence the
oxygen requirement, there are several physical, chemical, and physiological
conditions which influence the ability of fish to extract oxygen from the water,
its need for oxygen, and its ability to resist low oxygen levels. First, it
must be realized that ability to extract oxygen from the water and to resist
low D. O. levels varies with the species. It is well known that dogfish, carp,
and gar can survive at much lower D. O. levels than trout and several other
fishes. Some fishes are more efficient in the extraction of oxygen or their
blood is not as sensitive to the presence of COg'
The amount of oxygen required by fishes is determined in part by
activity. It is generally recognized that a man lying in bed does not breathe
3-S deeply or require as much oxygen as one digging a ditch. It has been re-
ported that from two to four times as much oxygen is required by a fish when
it is active as when it is quiescent (24) (26) (34). Under actual stream conditions
fish must maintain its position against the current, find) pursue, and
catch its food, avoid its enemies, and reproduce. All these activities require
oxygen in such amounts that D. O. levels at which the fish can just survive are
unsatisfactory. Age, size and season are also of importance. In general,
fry and younger fish have a higher metabolic rate and require more oxygen
than adults (35) (36). Because of increased activity and their physiological
condition fish require more oxygen at the spawning season. Studies carried
aut in our laboratories indicate that D. O. requirements are different at
ather times of the year and further, the physical condition of the fish is of
Outstanding importance in determining requirements and the minimum level
•olerated. An actively feeding, rapidly growing fish requires considerable
more oxygen than one which feeds very little. Since growth is rapid in the fry
•o fingerling stage it is expected that for many species D. O. requirements
_7*A_
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will be higher at this period. Eggs deposited in bottom materials require
higher D. O. concentrations than do adult fish. Since the current through
the bottom materials is slow, the amount of water flowing by the eggs per
unit of time is small and thus it must contain more D. O. to provide needed
requirements.
Through acclimation, resistance to low D. O. levels may be increased.
Fry (31) reports that through acclimation the lethal dissolved oxygen level
can be reduced to about one-half the corresponding value for trout accustomed
to air-saturated water. Lower dissolved oxygen levels can be tolerated for
considerable periods through an increase in respiration rate and volume of
water pumped, reduced activity and food consumption, and an increase in
blood haemoglobin (37) (38). By means of such adaptation fishes may live for
considerable periods at reduced oxygen concentrations without apparent harm.
This does not mean, however, that they can complete their life cycle at such
levels. Further, ability to live more or less indefinitely at low oxygen levels
does not mean that some of their physiological processes have not been altered
so that their well being and growth are adversely affected. It has been reported
(4) that the bullhead is unable to become acclimated to increased temperature
when D. O. levels are low whereas it becomes rapidly acclimated at normal
D. O. levels. Dissolved oxygen levels adequate for growth, reproduction,
normal activities, and well being are considerably higher than levels which
can be tolerated for extended jaeriods through acclimation and compensation.
Studies of the oxygen requirements of fishes fall into two categories:
laboratory investigations, where as many as possible of the variables are
controlled, the factor under study is varied, and the effects on fishes
directly observed for a relatively short period; and field studies, where the
variable in question is measured in different sections of the stream and is
related to the fish population in various areas. Both types of study have
certain advantages and disadvantages. It is very difficult to relate laboratory
results to field conditions, while in the field studies, factors other than the
variable in question (dissolved oxygen concentration) might have a bearing
upon results. It is believed that the best approach is to carry on both laboratory
and field studies so that they supplement each other. In the interpretation of
laboratory findings, it must be recognized that fish are usually held under
favorable conditions and it is necessary to realize that all findings are not
applicable to natural conditions.
The Lytle Creek studies (40) and other field studies have indicated
dissolved oxygen concentrations at which fish and their food supply can
maintain themselves. Twenty-four hour studies were made on Lytle Creek
at all seasons of the year at selected stations to determine D. O., CC^i pH,
and temperature. Such studies or a continuous record of dissolved oxygen are
essential for investigations of D. O. requirements as there are great
diurnal and seasonal variations in oxygen concentration. Fish populations
and growth rate studies were made over a two-year period (41) (42) in order
to relate them to environmental conditions and their seasonal variations in
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i.n umcicm ^uiuojia ui i~ne Biream, uncontrolled variables encountered in
stream studies usually make it difficult to be certain that differences in fish
populations are caused by oxygen concentration alone. However, it is be-
lieved that variations in oxygen concentration were the important variable
in Lytle Creek since fish appeared first in the riffles of the upper zone of
recovery and were found first in the pools much farther downstream. Since
fish were not present in the pool immediately below the riffles or between
them, it is believed this difference is due to D„ O, as other limiting factors
probably would not change so rapidly. In streams having a considerable
biological oxygen demand there are marked differences in D, 0„ in the pools
and riffles, In studying a section of the Scioto River, it was found that the
D, O, at the tail of a large pool was 0. 1 p. p. m. while about 200 yards down-
stream, water which had passed over a wide shallow riffle on one side of
the stream contained 5. 6 p. p. m. oxygen. Some 20 feet from the riffle in the
main flow of the river there was 2. 5 p. p. m. of oxygen.
In streams polluted with organic wastes, toxic materials such as H?St
NH3, and CH4 may be formed by anaerobic decomposition, The H2S may
escape or be fairly rapidly used by certain bacteria such as Beggiotoa,
Thiothrix, and Sphaerotilus (43). Usually much of the NH^ is converted to NC
and both of these materials are rapidly utilized by the dense growths of algae
in the recovery zone (44). Most of the CH4, which is not very toxic, escapes
a gas, Thus, while toxic materials may be formed, it is possible that they
do not exert a marked effect on the fish.
Determination of the oxygen requirements of fishes and establishment
of suitable dissolved oxygen criteria are especially difficult tasks, A great
many studies have been made of the oxygen requirements of fishes. Investigat
have not always used a uniform approach. In fact, there has been great
diversity in the species studied, the conditions under which they were studied,
the experimental methods used, the objectives of the study, the caliber of
the investigation, and the interpretation of results. Consequently, data ob-
tained have varied widely and have not always been in agreement. Short
time studies carried out in aquaria at low temperatures with resistant fishes
which are not fed indicate only that certain fishes can survive very low
concentrations of dissolved oxygen for limited periods. It should be re-
cognized that these levels are not adequate for normal existence or completion
of the life history of all the important fishes. In setting water quality
criteria for the protection of aquatic life, it must be recognized that mere
survival is not enough and that the minimum dissolved oxygen level should be
one suitable for the continuous maintenance of a satisfactory fish crop.
Minimum D. O. levels at which some species of fish can, through adaptation,
resist death by asphyxiation for a time are not adequate for completion of
the normal life cycle. Oxygen levels must be continuously adequate for the
general well being of the fish and the maintenance of fish food organisms.
Before adequate criteria can be established it is essential, therefore, to
know the environmental requirements of the fishes since the objective is to
provide suitable conditions for them.
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Concentration of dissolved oxygen is often expressed as weekly,
monthly, or sometimes daily, averages. Such values are not satisfactory
as they do not indicate environmental variations and may actually be mis-
leading from the standpoint of the continued existence of the fish. It is the
extreme variations which may become limiting and which are the most im-
portant for indicating unfavorable habitats.
Some D. O. criteria have been set up as percentages of saturation.
This procedure is deemed undesirable because over the range of temperature
observed in our natural waters, 50 percent of saturation may mean 7. 3 p. p. m.
oxygen or 3. 5 p. p. m. As temperature increases the amount of oxygen which
can be held by the water decreases, whereas the amount required by the fish
increases. It is believed that criteria for dissolved oxygen should be ex-
pressed in parts per million by weight.
Findings in Lytle Creek have indicated that in a stream section in
which the oxygen concentration is usually above 5 p. p. m. , the occurrence
of concentrations below 5 p.p. m. , but not below 3 p.p. m. for a few hours,
does not have an adverse affect upon a well rounded warm-water fish population
Minnows and other coarse fishes were found in the section where minimum D. C
levels dropped to 2 p. p. m. or slightly below. On the basis of these studies
and other pertinent data it is believed that for a well rounded warm.-water
fish population, dissolved oxygen concentrations must not be below 5 p. p. m.
for more than 8 hours of any 24-hour period and at no time should they be
below 3 p. p. m. For the maintenance of a coarse fish population dissolved
oxygen concentrations should not be below 5 p. p. m. for more than 8 hours
of any 24-hour period, and at no time should they be below 2 p. p.m.
The salmonoid fishes are not usually found in streams where minimum
dissolved oxygen concentrations are lower than 4 to 5 p. p. m. For normal
feeding and adequate growth at least 5 p. p. m. dissolved oxygen are required.
Successful development of eggs and fry require a minimum of 6 p.p. m. , while
for the full range of activity for brook trout and perhaps for other members
of the family, 7. 6 p. p. m. are required at 15°C. and full air saturation at
20°C. and above (31). It is believed, therefore, that for good salmonoid
production dissolved oxygen concentrations should not be less than 6 p. p. m.
Carbon Dioxide
Carbon dioxide may influence the toxicity of other materials or it
may in itself be harmful if present in sufficient quantities. Alabaster and
Herbert (48) found that CC>2 was not toxic to rainbow trout within a 12-hour
exposure at concentrations up to 30 p. p. m. but was toxic at 60 p. p. m. and
that period of survival decreased as the concentration increased. The
presence of CO2 in concentrations from 15 to 60 p. p. m. was found to reduce
the toxicity of ammonia. Higher concentrations are toxic; 100 to 200 p. p. m.
can be rapidly fatal to moderately susceptible fresh water fishes in well
oxygenated water. Fifty to 100 p. p.m. can cause distress and may be lethal.
Both marine and fresh water fishes vary greatly in their resistance to C02.
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Wells (49) reports that resistance of fishes to harmful conditions varies
with the species, with age or size and weight, with the condition or physiological
state of the individual; and with the season, He found that practically all
the fishes with which he worked were least resistant just after the breeding
season--June, July, and August--and most resistant before it--March, April,
May (50). Powers (51) has shown that the ability of marine fishes to extract
oxygen at low concentrations was adversely affected by moderate amounts of
CO2 which lowered the pH, The investigation of Black, Fry, and Black (52)
demonstrated the influence of CO2 on the utilization of oxygen by 16 species
of fresh water fishes, It was found that oxygen in the respired water at the
time of death was higher when the tension of CO2 was increased. The ability
to take up oxygen in the presence of CO2 varied with the species. Powers
and co-workers found (53) that fish are able to absorb oxygen at a low oxygen
tension through a wider range of CO2 tension than is found in the natural
waters in which they live. Most workers have found that naturally occurring
levels of CC>2 are not detrimental to fishes. It is believed that concentrations
under 30 p. p, m in the absence of other adverse factors will have no harmful
effects on most species, The majority of investigations indicate that CO-
becomes rapidly harmful at concentrations of 100 to 200 p. p, m. Surber
(54) found that concentrations between 55 p. p. m. and 78. 5 p., p. m. in hard water
at pH 6. 9 to 7. 0 caused a decided increase in the loss of eyed eggs and the
number of deformed trout fry. Concentrations up to 43 p. p.m. apparently
had no harmful effect.
Dissolved Solids
Natural, unpolluted waters of lakes and streams have in solution
small amounts of the anions CO3"", Cl SO4""", smaller quantities of NQj ,
NH4+ , P04 , and NC^". and traces of many others, The metallic
cations are Ca++, Mg++, Na+, K+, Fe+++, Mn*+, and traces of several
athers, These materials exert a physiological and osmotic effect to which
organisms have become adapted. In fact, these dissolved materials are
required by the organisms, Rawson (55) found a positive correlation between
:he total solids in fresh waters and the average standing crop of plankton
ind bottom fauna, The type of rock formation and soil largely determines
:he concentration of dissolved solids in a water but erosion may be of considerable
mportance Pollution may also be a factor, During the period from 1906-07
0 1934-43, the average amount of dissolved solids in Lake Erie increased
!rom 133 to 165 p,p.m,s whereas those in Lake Superior remained unchanged
56) (57),
Criteria for dissolved solids have little meaning if the purpose of
he criteria is the protection of aquatic life, unless the materials to be
:onsidered as dissolved solids are specified. It is apparent that salts of Hg,
I/u, Ag, Zn, Pb, and Cd will have a much different effect on fishes than will
jqual concentrations of salts of Ca, Na Mg, and K. In general when total dissolved
3olids are referred to in relation to water quality criteria, it is the salts
>f these relatively nontoxic earth metals which are believed to be under
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consideration. Unnatural concentrations of these salts may affect aquatic
life in two ways. If the solution of salts is physiologically unbalanced, one
of them may exert a direct toxic effect. If they are physiologically balanced,
that is, each is present in quantities sufficient to antagonize any toxic
effects of one or more of the others, they may occur in such concentrations
that they exert an osmotic effect. Wiebe (58) points out that the osmotic pressure
that fish can tolerate depends to a large extent on acclimatization. Fish
acclimated to the extremely soft waters of East Texas cannot survive when
subjected to salinities to which the fish of the Pecos River are continually
exposed. Texas rivers (58) range in total dissolved solids from 45 to 4, 810
p.p.m. Wiebe found as much as 28, 000 p. p. m. of chloride in a stream where
fresh water fish were supposed to live. However, Young (59) indicates that
when dissolved solids reach 11,000 p. p. m. only certain fish can tolerate
them indefinitely. The ability to resist high concentrations of dissolved solids
varies with the species. While some fishes move from marine to fresh water
or from fresh to sea water, some species have been reported as being unable
to resist concentrations above 3,000 p. p. m. Young reported that Na2CC>3
in concentrations about 800 p. p. m. was unfavorable for catfish. Huntsman
(60) reports that in the Quill lakes of Saskatchewan, which have a total solids
content of 16, 550 p.p. m. , there is a resident fish population of somewhat
limited extent. It is believed that total dissolved solids in concentrations up
to 3, 000 p. p. m. can be tolerated by most fishes if the materials in solution
are the nontoxic earth metals and are physiologically balanced.
Chlorides
The amount of chlorides is often considered as a measure of salinity
or of total dissolved solids. When dealing with sea water, which is fairly
uniform and the composition of which is known, chlorides can be taken as a
measure of salinity or dissolved solids. This does not hold, however, for
oil field brines and other wastes. Oil field brines differ drastically from one
another and from sea water and many wastes contain large quantities of salts
other than chlorides.
The chloride ion as such does not have much significance from the
standpoint of toxicity to aquatic life. The cation is so much more important
that the chloride anion is not generally considered. This is especially true
with chlorides of the heavy metals such as mercury, copper, zinc, etc.
Even with salts of the relatively nontoxic earth metals the toxicity of their
chlorides is evidently attributable to the specific toxicity of the cations present
and not to any toxicity of the chloride ions. Since the cations vary greatly
in their toxicity to fish and are governing in the determination of toxicity,
it is obvious that the chloride ion content of a mixture of salts is not a
reliable index of toxicity.
Physiologically balanced mixed salt solutions such as sea water may
be harmful to freshwater organisms because of their excessive over-all salt
content and osmotic pressure rather than the specific toxicity of any particular
ions present. Provided that the salts and other substances dissolved in water
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are balanced against each other so as to exclude any individual toxic effects,
certain hardy freshwater fishes can tolerate waters of osmotic pressures
equal to those of their own bloods and even higher for extended periods.
Other freshwater fishes, however, have been reported unable to tolerate
balanced salt solutions with concentrations of 2, 000 to 4, 000 p. p. m. It is
not known whether a typical freshwater fish can complete a normal life
history in water of relatively high osmotic pressure, nor is it known how
osmotic pressure affects the life processes of other freshwater organisms.
It is, therefore, impossible at present to define the maximal safe osmotic
pressure of a freshwater environment. Presumably, the tolerable osmotic
pressure entails a salt concentration far higher than the limits imposed by
industrial and municipal requirements. When we are dealing only with
different concentrations of a particular salt mixture such as sea salt; the
composition of which is known and uniform, the chloride content of the
solutions (which is easily determined) can be a useful index of osmotic strength.
However, when we are dealing with mixed salt solutions of unknown and varying
composition (such as oil well brines and other industrial wastes), their
chloride ion content is not a reliable index of osmotic strength. For example,
an industrial waste brine containing a large amount of sodium sulphate can be
much more active osmotically than another brine with a much higher chloride
ion content which contains only sodium and calcium chlorides. It is impossible
when dealing with mixed wastes to generalize as to the relationship between
chloride ion concentration and osmotic, toxic, or over-all pollutional strength.
It is believed, therefore, that chloride ion criteria have no practical
significance as far as aquatic life is concerned.
Fluorides
Studies at our laboratory in Cincinnati have indicated that fluoride
ions do have toxic properties in their own right. Further, they appear to
have a cumulative effect. In 10-day tests it was found that the TLm value for
potassium fluoride was 64 p. p. m. It is believed that for good fish-
production the fluoride content should not exceed 5 p. p. m.
Toxic Materials
There is a great deal of literature dealing with the toxicity of
various pure chemicals to fishes. The great majority of investigators
have used different approaches and have carried out their studies with a
variety of fishes, using different types of water for dilution. Several com-
pilations, reviews, or bibliographies of these studies have been made (38) (61)
(62) (63) (64) (67). An examination of these papers clearly indicates that
there is great variation in toxic levels reported by various investigators
for selected pure chemicals. This variation is especially evident in the
California report (61).
The quality of the receiving or dilution water, which is often not
reported by some of those investigating toxicity, is of outstanding importance
in determining toxicity of a particular material or waste. Several environmental
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factors may influence toxicity,, such as: temperature, CO21 D. O. , pH,
alkalinity, hardness, turbidity, and dissolved materials. Certain dissolved
materials may significantly affect the toxicity of a waste through their
synergistic or antagonistic action or through complexation, precipitation,
or other action. In the case of ferro- and ferricyanide solutions, sunlight
is of importance as photo decomposition of these materials occurs with
the production of toxic HCN (65). The heavy metals are considerably more
toxic at low pH since they are more soluble in acidic solutions. In hard
waters at higher pH they are precipitated or changed in other ways to become
much less toxic. It has been found that beryllium and uranium are 60 to 80
times more toxic in soft water than in hard water (66). Ellis (38) and other
investigators have found that ammonia becomes rapidly more toxic at pH
values above 8. 0. Calcium antagonizes the toxicity of many of the heavy
metals whereas some of them are synergistic with each other and become
considerably more toxic when mixed; examples are Cu and Zn, Cu and Cd,
and Zn and Ni (67).
The toxicity of many of the metal-cyanide complexes is greatly
influenced by pH. Doudoroff (68) has reported that fish can withstand 1, 000
times as much nickel cyanide complex at pH 8 as at pH 6. 5. Among the
metabolites, including weak acids and bases, it is the molecule and not the
ion which appears to be toxic. Thus weak acids such as HCN and
become more toxic as the pH is lowered and dissociation depressed, whereas
weak bases such as NH^OH become more toxic as the pH is raised.
In general, materials are more toxic at higher temperatures and at
low dissolved oxygen levels. Carbon dioxide may, through its effect on pH,
render some materials more toxic or it may serve to make others less toxic.
Complexation, precipitation, oxidation, dissociation, recombination, or
buffering action must also be considered. The influence of water quality and
other environmental factors on the toxicity of various materials has been
summarized by Tarzwell (69).
The character of the receiving water can cause wide variations in
the toxicity of many materials to fishes. Reference to the literature on
the toxicity of specific pure chemicals is of little value for determining the
toxicity of a complex waste containing these and other materials. Such an
approach neglects water quality, which is of particular importance, as
well as synergism and antagonism, oxidation, precipitation, complexation,
and other actions which may occur in the stream and may greatly influence
the toxicity of a waste in a particular stream. Numerical standards for
toxicity of specific pure chemicals have little value and can be misleading.
From the standpoint of industry they may be very undesirable. If a regulartoty
agency sets numerical criteria for the heavy metals and other substances
which are to be applied over a wide area, they must be set so low that allow-
able concentrations are-not detrimental to aquatic life under those
conditions at which they are most toxic. For example, criteria for
nickel cyanide wastes discharged into an acid stream would have to be set
so low that the HCN formed from the wastes would not be toxic, whereas in
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—. on caul me urueria couia be much higher as an increase of one and
one-half pH unite from 6. 5 to 8. 0 decreases the toxicity of this material over
1000 times. Copper is much less toxic in hard water than it is in soft water
with a low pH; variation may be as great as 200 times.
It is believed that the best approach to this problem is to make bio-
assays with the total waste, using for dilution, water from the receiving
stream at the point where the waste is to be discharged. In this way the many
variables which influence the toxicity of that particular waste in that stream ar<
taken into consideration and safe disposal or dilution rates can be determined.
With certain exceptions (some of the insecticides and other materials whose
toxicity is not influenced by water quality), water quality criteria for toxic
pnaterials when applied over wide areas should not be expressed as numerical
values. It is believed that a tailor-made approach should be used where
toxicity of the waste can be determined for the particular receiving stream
at the point of discharge. When the toxicity is determined in this way allow-
able concentrations for that particular situation can be expressed as p. p. m.
Dr as dilution ratios. Such an approach also permits evaluation and comparisor
Df the toxicity of wastes from different industries along the stream.
Summary and Conclusions
The establishment of water quality criteria for the protection of our
raluable aquatic resources is a complicated and difficult problem. Since the
»asic objective of water quality criteria for the protection of aquatic life is
0 provide or preserve environmental conditions essential for the survival,
ormal growth, reporudction, and well being of aquatic organisms a know-
edge of the environmental requirements of these organisms is essential for
be establishment of such criteria. Many of the activities of man have modified
tie aquatic environment. Among these are deforestation, unwise agricultural
ractices, overgrazing, and pollution. Our aquatic resources which produce
illions of dollars yearly in revenues from sport and commercial fishing are
renewable resource worthy of our best efforts for preservation.
Siltation due to erosion has been and is a major pollutant. The solution
f this problem by means of erosion control must be a cooperative effort
mong those agencies dealing with water, soil, and other natural resources.
While all the environmental requirements for aquatic life are not now
nown, application of the data presently available can be used effectively
1 the setting up of some criteria. As more data become available existing crite
an be modified and others set up in order to meet the problem adequately.
The quality of the receiving water is particularly important in determin
ie effects of many wastes. Among the factors influencing the toxicity of
sllutants in a particular receiving water are temperature, COz, D. O., pH,
kalinity, hardness, and dissolved materials. Other factors which may modify
e toxicity of a waste are complexation, precipitation, oxidation, synergy,
id antagonism.
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In view of the many factors which may influence the effects of
pollutants in different streams, it is believed that in most situations numerical
criteria when applied over wide areas can be set only for temperature, D. O, ,
and pH. With a few exceptions, such as for insecticides and certain other
materials, it is believed a tailor-made approach should be used in setting
criteria for toxic substances. This approach would consist of bio-as9yas of
the waste in question, using for dilution, water from the receiving stream
taken from the area into which the waste is to be discharged. Such bio-
assays would take into consideration the variables which influence toxicity
of that particular waste and can be used to indicate safe concentrations, or
the amount of dilution required.
Criteria for settleable solids and turbidity will (depend largely on
local conditions and will vary with the stream and the area.
When considering large areas pH values should not fall below 5 or
exceed 9. 5, but for good fish production it is desirable that they be maintained
between 6. 5 and 8, 5.
For a well rounded warm water fish population dissolved oxygen level!
should not be below 5 p. p. m. for more than 8 hours in any 24 hour period and
at no time should they be below 3 p. p. m. If a coarse fish population only
is desired minimum levels may fall to 2 p. p. m. For good production of
Salmonoid fishes a minimum of 6 p. p. m. appears to be required. However,
trout can live and reproduce in waters where the D. O. content may drop to
4 or 5 p. p. m. but the survival of eggs and fry and the production is usually
not so great.
It is suggested that in the northern portion of the country peak
temperatures for warm water fishes should not exceed 93°F. In the southern
portion of the country fish are better acclimated to higher temperatures and
can withstand peak temperatures of 96 F. or higher. While trout can stand
peak temperatures of 80°F. to 83°F. for short periods, best production is
attained in streams having summer temperatures of 60°F. to 68°F.
Allowable concentrations of toxic complex wastes for each stream or
situation should be determined by means of bio-assays and safe dilutions
estimated by the use of application factors or by other means.
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