f.
             ENVIRONMENTAL PROTECTION AGENCY
1                       REGION  VI
?                     DALLAS, TEXAS
         INVESTIGATION  OF FISH  KILLS
           BUREAU OF SPORTS FISHERIES AND WILDLIFE
               OKLAHOMA COOPERATIVE FISHERY UNIT
                  OKLAHOMA STATE UNIVERSITY
                    STIL LWATER, OK LAHOMA
                        APRIL, 1972

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                                      Collection of Papers
 M                                      presented at the
 •                               FISH KILL INVESTIGATION SEMINAR
                                               on
 I                                     November 2-4, 1971

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                                      Sponsored Jointly By
 •                               Environmental Protection Agency
 •                                          Region VI
                                          Dallas, Texas
 I                                             and
                                Oklahoma Cooperative Fishery Unit
 I                                  Oklahoma State University
 •                                     Stillwater, Oklahoma

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                                           April 1972
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                            FOREWORD
Floating dead fish have always been an indicator to the observer
that something is not right with the environment.  Since fish
mortalities may result from a variety of causes, some of natural
origin and some man-induced, a prompt, well organized investigation
is necessary in order to provide the information needed to determine
the cause of a fish kill and the source of the killing agent.

A fish kill investigation can be one of the most complex procedures
encountered by agencies concerned with water pollution control.
With the usual demand for prompt results, limited opportunities
for obtaining proper information, the high degree of variability,
and the many unknowns, a fish kill investigation can become a
perplexing and frustrating task.

Therefore, Region VI, Environmental Protection Agency and the
Oklahoma Cooperative Fishey Unit, Oklahoma State University,
cosponsored a seminar dealing with various phases of fish kill
investigations.  Federal, state, local and industrial personnel
from Region VI and nearby states interested in fish kills and
their investigation were invited to participate.  The purpose
of the seminar was to disseminate and exchange knowledge and
ideas concerning various phases of a fish kill investigation.

In this manual we have included a collection of papers that were
presented during the seminar.  In a few cases, papers could not
be provided by the speakers.  For the purpose of consistency and
clarity minor editorial changes were made in some of the papers
by the Seminar Director.
                                John E. Matthews
                                Seminar Director
                                   Region VI
                        Environmental Protection Agency

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                         CONTENTS


                                                                 Page
                           INTRODUCTION


Remarks to the Opening Session of the Fish Kill Investigation      1
Seminar.  November 2, 1971, Oklahoma City, Oklahoma
     Kenton Kirkpatrick


A Preface for the Investigation of Fish Kills                      4
     Kenneth M. Mackenthun


Rules of Evidence                                                 10
     Tony Anthony


State Legal Considerations                                        17
     Robert H. Mitchell


Steps in Establishing Good Public Relations with the              21
Media and the Public
     Diane Norvell


Alabama Program for the Investigation of Fish Kills               24
     Sam L. Spencer


Arkansas Fish Kill Investigation Program                          28
     Neil Woomer


Investigation of Fish Kills in Missouri                           31
     Frank Ryck


               NON-TOXICANT RELATED FISH MORTALITIES


Non-Toxicant Related Fish Mortalities                             33
     Robert C. Summerfelt


Natural Variations in Fish Populations                            37
     Austin K. Andrews


The Role of Diseases in Fish Kills                                46
     Fred P. Meyer


Mortalities from Miscellaneous Causes and Important               61

Environmental Considerations
     Ralph M. Sinclair

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                                                                 Page


                    TOXICANT CAUSED FISH KILLS


Toxicant Caused Fish Kills - Collection, Preservation,            70
and Analysis of Inorganic Samples
     Mark Coleman


Toxicant Caused Fish Kills - Collection, Preservation,            77
and Analysis of Organic Samples
     Larry Streck


Collection, Preservation, and Analysis of Organic Samples         81
     Robert E.  Reinert


Interpretation and Use of Data                                    83
     John E. Matthews


                ASPECTS OF FISH KILL INVESTIGATIONS


Fish Kill Investigations and Techniques                           87
     Ed Sorrels


Kill Number Estimates                                             99
     Byron Moser


Value of Fishes                                                  105
     Sam L. Spencer


Other Economic Damages                                           109
     Gene F. Forbes


The Fish Kill Investigation Report                               112
     John E. Matthews


            SPECIFIC INVESTIGATIONS AND CASE HISTORIES


Investigation of Petroleum Caused Fish Kills                     115
     Sterling L. Burks


Toxicant Caused Fish Kill in Alabama - A Case History            119
     Sam L. Spencer


An Endrin-Caused Fish Kill in Arkansas - A Case History          124

     Neil Woomer


Pollution Caused by Strip Mining in Missouri - A Case History    129

     Everett H. Fuchs

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  REMARKS TO THE OPENING SESSION OF THE FISH KILL INVESTIGATION
       SEMINAR.  November 2, 1971, Oklahoma City, Oklahoma

                               by

                       Kenton Kirkpatrick

           Assistant Director, Air and Water Programs
    Environmental Protection Agency, Region VI, Dallas, Texas
It is my pleasure to be here this afternoon representing the
Acting Regional Administrator of EPA, Region VI, Mr. Bill V.
McFarland.  Mr. McFarland asked me to relay his regrets to you
that other committments prevent his being here himself.  He is
participating in the Galveston Bay Enforcement Conference in
Houston.

So on his behalf and on behalf of all of EPA, welcome to the
Manpower and Training Program's Fish Kill Investigation Seminar.

I feel that the Fish Kill Investigation Program is a vital part
of the overall EPA operation.  Statistics alone support this view.
EPA's Annual Fish Killed Report for 1970, which is now at the
printer, states that there were 634 recorded incidents in the
United States involving more than 23 million fish.  That represents
a  36 percent increase over 1969 figures.  The 1970 totals bring
to 161 million the number of fish killed in 4,500 incidents
throughout the country since reporting began in 1961.

This shocking mortality rate provides us with convincing evidence
of the degradation of our Nation's waters.  It also stresses the
importance of investigating each incident thoroughly to determine
the nature of the lethal pollutant.

Unfortunately, all too often the cause of the fish kill remains
undertermined due to inadequate channels of reporting and methods
of investigation.  The EPA is only now beginning a serious effort
to establish an effective program in cooperation with State and
other Federal officials and concerned citizens.

During the next two days you will be discussing some of these
deficiencies and the techniques for correcting them.

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The EPA is involved in fish kill investigations on two levels.
The initial concern is with the field investigation conducted by
the Surveillance and Analysis Division.

When we are advised that a fish kill has occurred, a team of
technical experts is dispatched immediately to the scene.  But
inaccurate and untimely reporting too frequently prevents effective
investigation.  The team obviously cannot function effectively when
the cause of the kill has been washed away.

In an effort to improve the notification system, a program is being
developed in Louisiana on an experimental basis.  A special telephone
has been installed in our Baton Rouge Facility and will be manned on
a 24-hour basis to receive reports of fish kills.  I might as well
give you the number now because we are going to publicize it far
and wide.  It is area code 504, number 926-0188.

This number will be provided to all State and local government
agencies which have the potential for obtaining knowledge of fish
kills.  Fishermen and other private citizens who might be aware of
fish kills also will be provided with the number.

When they call, they will be asked a series of questions designed
to obtain the basic information.  When appropriate, a field team
can then be sent out.

Also tied into the system will be the very effective early warning
network established several years ago along the Lower Mississippi
River by the Louisiana Department of Health and Stream Control
Commission.  This network is designed to alert operators of drinking
water treatment plants taking water from the River when spills of
dangerous pollutants occur.  The in-take valves are then closed
until the contaminated water has passed.  Dead fish are a reliable
indication that there is trouble.

It is hoped that this system will enable our field teams to reach
the scene in time to evaluate the circumstances and determine the
cause.  This will make our reports, which are relayed to the States,
more valuable.

Our analysis attempts to pin down the specific material involved
and the source.

At this point our second-level concern is activated.  When it is
deemed necessary, the report is turned over to the Enforcement
Division for whatever action is indicated.

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In the event that legal action is to be taken, it is essential
that the data be reliable.  This again emphasizes the importance
of the investigation.


What is the relationship between EPA and the States?  The basic
foundation of this relationship is established in the Federal
Water Pollution Control Act.  This law places the primary respon-
sibility for pollution control in the hands of the State agencies.
So as far as possible, EPA relies on the States to enforce water
quality standards.


However, since expanding use of the 1899 Refuse Act, independent
action by the Federal Government has grown.  Pending Legislation
would give EPA direct enforcement authority over all navigable
waters rather than just interstate waters as at present.


When EPA proceeds on its own initiative, the State is kept fully
informed of our intentions and activities.


Whether it is the State or Federal Government taking the lead,
cooperation is essential to a successful program.  But no matter
how much cooperation is developed among the various concerned
agencies, the program can only really be successful if the
information about the incident is precise.


So we regard this seminar as not only an opportunity to exchange
knowledge and ideas but as an opportunity to develop and improve
our professional skills.


So again, welcome to the seminar and best wishes for a successful
program.

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          A PREFACE FOR THE INVESTIGATION OF FISH KILLS

                               by

                      Kenneth M. Mackenthun

         Acting Director, Division of Technical Support
      Water Quality Office, Environmental Protection Agency
Since the beginning of water pollution investigations, floating dead
fish have been a red flag to the observer that something is not right
with the aquatic environment.  Early in legislative history for water
pollution control, many States adopted particular laws to assess
damages from those who, through the introduction of foreign materials,
would render the States' waterways toxic to fish and other aquatic
life.  Fish kills have been the subject of litigation in the courts
and many times have served a useful purpose to stimulate and focus
public concern and attention towards appropriate water pollution
control.

The Federal government began a voluntary annual census of fish kills
in June 1960.  These pollution caused fish kills are reported
through appropriate State officials to the Federal water pollution
control authority (now the Water Quality Office within the Environmental
Protection Agency) where the data are tabulated and analyzed.  Since
1960 over 144 million fish have been reported killed in 4,200
incidents.

In 1969 alone an estimated 41,000,000 fish were reported killed In
45 States by identifiable pollution sources.  Of the number of reports,
industrial operations ranked first as a killer of fish, followed by
agricultural, municipal, transportation and other operations in that
order.  As expected, the months of May through September ranked
highest with the number of reported fish kills.  August was the
highest month with number of reports and December was the least.
Indicative as these annual summaries are, there remained an urgent
need to determine the specific cause of a particular kill so that the
cause could be corrected and water quality enhanced in the future.

In a news release dated August 21, 1969, Commissioner Dominick of the
Federal Water Pollution Control Administration said that the present
voluntary program of reporting fish kills is no longer adequate in
our stepped up overall campaign against water pollution.  The old

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system did not provide for immediate reporting of fish kill
incidents and did not require any counter action by the FWPCA.
Because fish kills indicate a violation of water quality standards
or enforcement conference recommendations, it is particularly
important that any sizable fish kill be reported and scientifically
investigated.  The Commissioner directed each Regional Director to
take an active role in investigating fish kills, in determining
their cause, in providing technical assistance on any immediate
control measures or warnings that can be taken and in providing
technical or other authorized assistance in designing control
measures that would prevent a recurrence of the kill.  Under this
plan the Regional Offices of the FWPCA were to establish procedures
with State water pollution control agencies to insure that they
receive immediate reports of significant fish kills within each
region.  It is recognized that the primary responsibility for the
investigation of fish kills rests with the States as is the case in
all water pollution control matters.  The Water Quality Office of
EPA shares these responsibilities in interstate waters or in waters
where there are Federal-State enforcement actions.

The proper investigation of a fish mortality is through a State-Federal
partnership.  The role of the Federal government is to offer technical
assistance to the State in the investigation of kills to the extent
that resources permit.  In areas of Federal jurisdiction we are
prepared to initiate independent action to achieve a swift and
adequate determination of the cause of a kill and of the appropriate
control and preventative measures if such action becomes necessary.
Fish kill coordinators have been identified for each of our regional
offices and in Headquarters.  The coordinators are in a position to
respond to requests for technical assistance from the States in any
aspect of the fish kill investigation and to receive the immediate
reports from the States on significant fish mortalities.

Obviously each fish death can not be investigated fully.  Only
significant fish kills should be reported.  The definition of
significance is a matter of judgement, but it should include
mortalities that are important because of sport or commercial fish
values, that result from a suspected negligent discharge or
malfunction of a waste treatment device, or that show widespread
environmental damage.

The immediate report received by the region and supplied by the
State should include sufficient pertinent information to identify
the fish kill area, the relative extent of the mortality, the kinds

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of fish Involved, whether or not technical assistance to investigate
the cause has been requested, and who should be contacted to obtain
additional information.  This information is then transmitted to
Headquarters by the Regions.

Follow up information developed in fish kill response investigations
or otherwise acquired should be sent to the Regional coordinators
and in turn to Headquarters of the Water Quality Office, as soon as
possible.  We are particularly interested in information concerning
the cause of the kill and actions to alleviate the cause, to collect
damages, and to prevent future incidents.  To make this fish kill
investigative partnership effective, we need to receive information
on fish mortalities immediately, by telephone.

Recently we evaluated our records relating to the program of reporting
and responding to significant fish kills.  In this evaluation we
choose a fisherman year - that period of time from September 1, 1969
to October 1, 1970.  At the onset I will say that our efforts have
not been enough and they must be improved.  In this period of time
231 fish kills were reported to my office under this program.  These
reports accounted for approximately 50,000,000 fish; and 860 miles
of stream and 10,700 acres of other waters were affected.  Of the
231 reported incidents, 194 were investigated in some fashion.  The
States investigated 170 incidents and the Federal government assisted
the States in their investigation of 21 incidents.  There is some
indication in the record that the States have requested and received
more assistance from the Federal government in the investigation of
fish kills during the latter half of our fisherman's year than
during the first half.  We hope that as this program matures and
develops that assistance from the Federal government will be
requested in a much greater number of investigations and that we
will be able to provide the type of assistance that will be
meaningful and definitive in determining the cause of fish deaths.

Initial laboratory support for the investigation of fish mortalities
should come from any Water Quality Office laboratory within the Region
where the mortality occurs.  If this support is unable to respond to
the needs of a particular investigation, additional special services
can be obtained from the Analytical Quality Control Laboratory in
Cincinnati, Ohio, the National Water Quality Laboratory in
Duluth, Minnesota, and the Southeast Water Laboratory at Athens,
Georgia.  Analytical capabilities in pesticides and other organics,
heavy and trace metals, oil identification, complex organic compounds
and certain fish diseases and bioassay analyses are available at
these laboratories.

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The Bureau of Sport Fisheries and Wildlife maintains a small staff
of disease specialists within the Division of Fish Hatcheries to
provide diagnostic services to national fish hatcheries. As time
permits, these hatchery biologists are available to assist other
agencies in diagnosing fish diseases.  Emergency inquiries for
assistance may be directed to the nearest hatchery biologist.  Such
hatcheries are located in the States of Washington, California,
Montana, Utah, Arkansas, Wisconsin, North Carolina, West Virginia,
and Maine.  In addition, there are two Bureau research laboratories
that specialize in fish diseases.  The Eastern Fish Disease
Laboratory, Kearneysville, West Virginia  25430 serves the area
east of the Mississippi River, and the Western Fish Disease
Laboratory, Building 204, Sand Point Naval Air Station, Seattle,
Washington  98115 serves the Western States.  Except in emergency
situations, requests for fish disease diagnostic services should be
directed through Headquarters of the Water Quality Office.

As far as the art and science of fish kill investigations are
concerned, one might say with truth that we are entering the age
of the Renaissance.  Historically we have advanced slowly and
simetimes it seems but little since the days of Shelford, Richardson,
Forbes, Juday, and Birge.

One of the greatest deterrents to the conclusive investigation of
fish mortalities is our seeming inability to arrive at the scene soon
enough.  Speed is all important in the initial phases of any fish
kill investigation.  Early arrival at the scene, after a fish kill
begins, usually determines the success of the investigator's
evaluation of the problem.  Observations of the actions of moribund
fish, and the collection of appropriate samples at this time, are
often a vital link to the arrival at a conclusive determination
of the cause of death.  Dead fish disintegrate rapidly, especially
in hot weather and the cause of death may disappear or become
unidentifiable within minutes.  Thus, the system of early warning
followed by speedy investigation is essential to a successful fish
kill investigative program.

The degree of success of an investigation will depend to a large
extent on the investigative preparedness of the party involved.
Planning is an essential component of any successful enterprise and
the successful investigation of fish mortalities is not an exception.
Planning will decrease the interval of time between notification and
arrival at the scene.  Appropriate planning will insure the
availability of proper equipment and testing facilities and it will
decrease the confusion and uncertainty that sometimes is associated

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with the analyses of collected samples.  Planning will identify
specific laboratory capabilities before the need for these occur.
Planning will alert reserve resources should these be required by
a particular incident.

The investigator should be ever mindful of the potential legal aspects
of his investigation.  The investigation should be conducted to
insure that collected data will be acceptable to the courts and will
withstand critical cross examination.  Samples collected in the field
and transported to a laboratory should be sealed and so identified
by the sender that the chain of evidence always remains intact, and
that it cannot be broken by opposing attorneys.  When the samples
are received in the laboratory they should be associated with a
continuous log that records the date and exact time of each step
in the analytical procedures and the person responsible for the
samples during that step.  When not in use, samples should be
retained in a locked room.  As an effective tool of the water
pollution control program, the evidence collected by the investigator
must withstand the scrutiny of the courts.  Preparation of the
investigative data in this manner will be an asset even though
remedial actions and pollution abatement procedures are arrived at
through informal negotiations rather than court action.

There are about six essential steps connected with the investigation
of a fish mortality.  The first is the collection of preliminary
information to identify the problem and its extent in very specific
terms.  There follows three concurrently attacked phases that
include physical observations, the collection of chemical data, and
the collection of biological samples.  Observations by an experienced
investigator are invaluable to define the cause of a given kill.
Perhaps no single factor is so important.  Particularly noteworthy
are the general appearance of the water, its color, clairty and
whether sparkly or dull and lifeless.  The presence of oil, excessive
algae, unusual biotic growths such as sewage fungus, excessive
amounts of higher aquatic plants and the color and appearance of rocks,
Sticks or debris within the water are important considerations.  Notes
on the actions of fish prior to death are important, as well as
observed actions of other organisms such as crayfish and other benthos.
Often much can be learned regarding the severity of the kill to the
aquatic environment by lifting a rock from the stream bed or a piece
of debris from the water and noting the types of organisms that
remain alive.  The fifth component of an investigation is the analyses
of the many collected samples.  The last embodies the tabulation and
interpretation of data and the preparation of the report.  These are

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equally important aspects of an investigation and, if it is to be
successful, all must be performed with great care.


The collection of chemical and biological samples and data involve
a number of technical considerations.  The Division of Technical
Support of the then Federal Water Pollution Control Administration
published a 21 page booklet entitled, "Investigating Fish
Mortalities."  This publication considers some of the technical
aspects of the sample and data collection.  It may be obtained from
any of our regional offices or in Headquarters.   I commend it to
your reading.

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                         RULES OF EVIDENCE

                                by

                           Tony Anthony

           Enforcement Specialist, Enforcement Division
     Environmental Protection Agency, Region VI, Dallas, Texas
The importance of evidence cannot be overemphasized.  The procedure,
the investigation, the methods, everything you will be hearing
today, everything you probably studied about, everything you will
learn in the future is all directed to the one key point of
evidence.

In response to the question:  how significant does a fish kill
have to be before you go out and collect good evidence?  If it is
worth investigating, it is worth documenting.  Those cases that get
to court are frequently the ones with the lousiest information
possible;  the one where some guy goes out and looks at it over the
weekend and didn't take any samples or didn't take any notes.  When
the case gets to court the lawyers are sort of hard put to try to
make something out of it.   So if it is worth investigating at all,
it is worth doing a good job on it.

What is evidence?  Evidence is really everything; a mass of
information and data which is prepared, preserved, presented to a
trier of fact in order for him to supposedly arrive at the truth
of various allegations or charges.  Now when I say it is everything,
naturally I have to immediately start making exceptions.  There are
probably as many exceptions to what evidence is as there are rules
trying to describe it.

One of the most well known exclusions of any evidence that I would
point out is hearsay evidence.  Here again there are numerous
exceptions to the so called hearsay evidence rule.  Basically,
hearsay evidence is something that you do not know or something
that cannot be presented as a hard, cold fact.  For instance, I
can see a container sitting on the front table.  I can testify to
the fact that there is a container sitting there.  One of the
gentlemen sitting there could tell me it was a container full of
water or a pitcher of water.  At the present time that would be
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hearsay evidence as far as my testifying to the fact that there is
a pitcher of water on the table.  Here again I would immediately run
into one of the exceptions to the hearsay rule.  If the gentlemen
who told me there was water in the pitcher sitting on the front
table is also in court and can rebutt my statement, I can testify
to the fact that that is water in the pitcher.  But if he is
nowhere around, and I just say something that someone else told me,
it is excluded as admissible evidence.

What are some of the forms of evidence?  Most of it, a good deal
of it, is oral testimony or personal knowledge of a witness,
someone who has seen or knows from his own personal knowledge about
a particular fact.  If I further investigate the container sitting
on the table, it looks like water, it sounds like water, and it
probably feels like water.  But it may be gin, so I can't, at this
point, testify that that is water in that pitcher.  Now if I pour
a glass of it, drink it and it is water, that is evidence that I
can testify to.

Circumstantial evidence is used to a considerable extent by lawyers.
Mainly because we do not have the factual evidence that we really
need to tie one point to another.  We can string a group or a line
of individual facts that will lead to a particular assumption or
interpretation.  My interpretation right now is that there is water
in the pitcher.  Conceiveably, what I have seen here today could
be circumstantial evidence that there is water in the pitcher.

We also have graphic evidence.  Photographs are excellent examples
of graphic evidence.  If you are in an investigation of a particular
water body that looks to you to have caused the fish kill, horrible
looking, real bad stuff, get a picture of it.  You see fish floating
in it belly up, get a picture of it.  Very recently in a federal court
case in which I was participating, one of the witnesses had color
slides of a particular body of water.  He had put dye in this water
and traced it down the waterway, taking pictures now and then.  At
one point it just so happened the dye was in the same field of view
of his camera as three or four floating dead fish.  He was projecting
these slides on a screen for the judge.  The minute that picture of
those dead fish hit that projector the judge stopped everything,
gently folded up his robes, and traipsed down to where he could get
right in front of that screen and see the dead fish.  This is beautiful
evidence, it is hard to refute.  Now there are problems of tying  the
evidence in with what caused their death.  If you can document with
photographs what you are investigating, your attorney can probably
tie this into a pretty effectual presentation.
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Demonstrative evidence is occasionally used in court.  This ordinarily
is within the discretion of the court itself as to whether it will be
permitted or not.  If we had, for instance, a case where it was
suspected that ammonia was the source of death in a particular fish
kill, conceiveably you could bring a small tank of fish in their
natural water into court and insert a given amount of ammonia to
demonstrate that these fish will be killed, or their gills will start
swelling and bleeding.  Be sure and check with your attorney so he
can check with the court before this type of evidence is prepared
or anticipated being used.

Another thing I want to point out is that one of your prime functions
in this evidence area is educating your attorney.  Some attorneys
aren't exactly right up on their toes on all this technical jargon.
You may have evidence coming out of your ears, but you must get over
to your attorney and educate him as to the value of this evidence.
It really is not going to do any good if he can't get it to the
trier of facts.

Another particular type of evidence which may be considered is derived
evidence, or calculated evidence.  When a particular stream or water
body is sampled and this sample taken back to the laboratory and
analyzed, certain calculations may have to be made, e.g., it has to
be measured against certain standards.  The end result of these
manipulations is more or less a calculated piece of evidence.  You
can demonstrate the reliability of the evidence if your procedure
and method is reliable.  If the very fact which you are trying to
present is not so self evident that you can bring it in and set
it on the court table, you must calculate something to show the
trier of fact what you are trying to present to him.

Most witnesses in a court case such as a fish kill case probably
will be just plain, straight, factual witnesses.  An investigator
who counts the fish and calculates the number of dead fish, who
samples the water, who has taken photographs, can testify to the
fact that he took the photographs, he sampled the water, he counted
the fish.  He may not be expert enough in a particular area to
determine what caused the death of these fish.  This is where we
arrive at the role of an expert witness.

An expert witness is an expert in his given field.  An ichthyologist
for instance may be a specialist in a particular phase of the study
of fish.  He may have studied a considerable amount on the cause of
death of certain fish.  This man, once he is qualified by education
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and experience, is allowed to make an interpretation from other
factual data which he may not have personally picked up himself but
which has been presented to the court.  For instance, the investigator
established as a fact that he has sampled a given water body, he
has picked up dead fish and he has turned the samples over to a
chemist.  The chemist gets on the stand and testifies to the fact
that he analyzed this particular sample and found x, y, z parameters
in it.  An expert witness could then get on the stand and from this
information could make an interpretation to present to the court
that these x, y, z parameters were probably the cause of death of
these particular fish.  An ordinary lay witness is not allowed
to make an assumption or interpretation of this nature.  An expert
witness in biology is not permitted to make an interpretation in
the field of chemistry, a field where he has no expertise.

The problem of what evidence is admissible is also one with which
we have a struggle.  In every case that comes up something is going
to be objected to as being admissible or not admissible.  Here again
this is not primarily your function, but you should be aware enough
of the problems the lawyer faces to be able to help him.  As I
mentioned before you may have a trunk full of data, but unless you
can explain the significance of this data to the attorney who is
handling the case he may not know whether that trunk full of data
is admissible or not.  In order for it to be admissible it must be
at least relevant to the issue before the court.  By relevance we
might illustrate with the quality of water in a stream in which dead
fish have been found.  This is probably relevant to a fish kill because
of the fact that fish ordinarily live in water.  It may or may not be
material if the fish are dead because they were seined and left on
the bank to die.  The quality of the water may be completely immaterial,
if the quality of the water shows, for instance, certain parameters
of a, b, c in the water, and the fish died from x, y, z causes.  The
amount of a, b, c in the water may be completely immaterial to the
issue of where the x, y, z came from or what caused the end result or
death from x, y, z.  You can't just throw in everything you know about
the water quality.  If you found a dead horse in the stream, it may
be relevant or material to the issue; it may not.  You have to use a
little of your judgement, and you have to help educate the attorney as
to the particular significance of the information you are trying to
get to the court.

Once a sample of water has been taken to a laboratory to be analyzed,
those of you who are doing this analyses must always use a scientifically
recognizable test in order to establish what was in this particular
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water.  If you have dreamed up a test all of your own when everybody
else in the world is using standard methods, it may be the best test
in the world but unless it is widely accepted within the profession,
it will probably go to the weight of the evidence whether the trier
of fact should accept it or not.  So the results of your work must
depend upon the acceptance by the scientific discipline in which
you are endeavoring to establish these facts.

The chain of custody of samples is a very important part of any
investigation, and it can't be expressed enough how important it
is.  If you are an investigator and you collect a sample of water
from a particular stream or waste effluent, identify each sample
as you take it with the date, the time of day, any witnesses that
might have been there the same time and hopefully you have some
witnesses, whether there was any preservative put in the sample,
very definite information that would identify this particular
sample as being taken from that particular spot.  Each individual
sample must be identified.  Don't collect 25 different samples up
and down the stream and them put the same tag on all of them.  It
can completely invalidate the results of any particular one.

Once you have picked up a sample, establish a regular, orderly
procedure for transferring that sample to your analyst  or to your
lab.  If you are doing it yourself establish methodical procedures
for handling the sample.  Do it that way every time.  There may
be several different ways of transferring this material, or sampling
or even analyzing and they may all be equally valid, equally acceptable,
but any one particular organization is going to be much better off
in a court of law if they can establish that they have done this
particular job the same way every time.  For instance, I go out to
take a sample of water, the parameters I need it tested for make it
mandatory that I preserve this sample before I leave the site and
take it back to the laboratory.  It may be immaterial to the analysis
of that sample whether I put the preservative in my container before
I left the lab or whether I put the preservative in immediately after
I have filled the container with sample.  Assuming it is equally valid
one way or the other, establish a procedure whereby you either always
put the preservative in the container prior to sampling or you always
put it into the sample after it has been taken.  If one day you do it
one way and the next day you do it the other, it is not too difficult
a task for an attorney on the adverse side to establish that maybe you
put in an extra dose or that you failed to put in any.  This can tear
up some awfully good evidence.  So establish a set pattern.  Do it
the same way every time.
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Once these samples have been collected keep a chain of custody tag
with that sample at all times, with its identification on the outside
of the container.  Once you turn the sample over to your analyst,
sign off on the tag, date it, put the time on it that you turned it
over to this particular individual, have him sign off on it that he
received the sample at that particular time.  Then when he turns the
sample over to the fish chemist this procedure must be followed
through on the chain of custody.  Each time the sample leaves one
person's hands or custody it should be recorded so that the attorney
can trace this sample all the way to the very final results, without
having a gap somewhere.  Once a gap can be found in a chain of custody
it can tear hell out of the results, and you can lose the case.  And
that is not the name of the game.

If you must, in the transporting of the sample, turn it over to a
common carrier, for instance an airline, have the agent of the
carrier sign off on the custody tag.  Have your custodian at the
other end sign that he received the sample.  You can't be held
responsible for everything the agent does with it.  There may be
numerous other people who handle it in between, but that is not
your responsibility.  That is the responsibility of the common
carrier, and the attorney can get to the root of that.  He can
bring the common carrier into court to establish the custody
during that period of time.

Once you have analyzed and completed the work on your sample, don't
toss it down the drain.  Save enough of the sample so that it can be
analyzed again for a duplication of results, in case it does come
to court.  Now I realize this causes trouble.  There is not enough
warehousing space anywhere in the country to house all the samples
that have been taken.  You can't keep them forever, but you can keep
them or a usuable portion thereof, for a sufficient amount of time,
before you destroy or throw them away.  The amount of time will vary
somewhat with each type of sample.  Some types do not lend themselves
to storage.  So please establish some procedure for retaining samples.
The sample that you throw away is frequently the one that is essential
in the one case you are called on, and that is the one your attorney
is going to be scratching his head trying to figure out, what do we
do now?

The main enforcement responsibilities of the Environmental Protection
Agency are related to the Water Pollution Control Act of 1965 and
amendments.  We do have several responsibilities set forth in the
Act, one of which :".s the conduct of enforcement conferences in given
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situations.  Another is to file civil cases against water polluters
under other given situations.  The enforcement responsibilities of
the Agency itself is to gather information or evidence and prepare
it to be presented to the Justice Department, which presently
handles all litigation for our Agency, in such a fashion that it
is usuable and is as close to a sewed-up case as you can get.  The
Enforcement Division of the EPA has the final say as to whether
an enforcement action will be undertaken or not.  Surveillance and
Analysis has the discretion of deciding whether there has been a
significant enough set of circumstances to warrant, in their opinion,
an enforcement action.  They will at that time forward this particular
incident to the Enforcement Division for review, and further action
if it is an acceptable case.

Also, as you are well aware, EPA has the responsibility, not for
enforcing the 1899 Refuse Act, but for preparing evidence and
determining the water quality, and the extent of refuse which
might be a violation under that particular Act.  It is up to the
Department of Justice to actually make the final determination
whether a prosecution under that particular Act will be pursued
or not.  Once they get that determination they can take either
civil or criminal action.
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                   STATE LEGAL CONSIDERATIONS

                               by

                       Robert H. Mitchell

                   Assistant Attorney General
                        State of Oklahoma
The Attorney General's office is currently trying the first fish kill
case in the State of Oklahoma in Payne County concerning a fish kill
on the Cimarron River.  We are unable to speak directly to all of the
issues since many issues remain unresolved and have not been decided
by the Court.  Eventually the jury will decide the issue once the
case goes to trial.

                      HISTORICAL BACKGROUND

One.  Case of State v. Wheatley, which was one of the early pollution
cases, wherein the defendant was charged by indictment and information
under the criminal statute which provides one year imprisonment and
$100 to $500 fine.  The appeal was to the Court of Criminal Appeals,
1921.  The case is significant because the Court recognized the
Constitutional rights of the State to protect fish and wildlife under
the policy power of the Constitution.  They found that it was valid
to charge an individual for the killing of fish under our Constitution.

Two.  Pollution in Oklahoma has been a problem through the oil
industry since it has been our major industry.  In effect, there
used to be associations of oil producers who would bind together
and obtain easements from all land owners surrounding streams and
lakes and oil fields and obtain from them, in effect, an easement
to pollute.  The land owner would sign an instrument, in return for
money, stating that he would forego any legal action he might have
in case of damage by salt water or crude oil or other deleterious
substances.

Three.  There of course are other cases, civil in nature, where
individual land owners can sue for damage to domestic animals or
property due to pollution.

Four.  In 1968 the next move was made by the Oklahoma Legislature
in setting up the Pollution Control Coordinating Act.
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The Oklahoma Statutes, 82 O.S.   931 to 942 hold the composition
of the Department of Pollution or the Pollution Coordinating Control
Board.

         PERTINENT STATUTORY AUTHORITY FOR LEGAL ACTION
                       IN FISH KILL CASES

Title 82, O.S.   931 to 942 - Organization of a pollution control
coordinating board and legal enforcement measures.

Composition of the Department of Pollution or the Pollution Control
Coordinating Board, nine members, (a) Commissioner of Health,
(b) President of the State Board of Agriculture, (c) Director of the
Water Resources Board, (d) Director of Department of Wildlife
Conservation, (e) Chairman of the Oklahoma Corporation Commission,
(f) Director of Industrial Development and Park Department, (g)
Executive Director of Soil Conservation Board, and (h) Two members
appointed by the Governor with the advice and consent of the
Senate ...  (i) the Attorney General of the State of Oklahoma shall
serve as the legal counsel for the Department of Pollution Control
and shall assist the Board in the performance of its powers and
duties designated by the Act.

First example of legal authority in 932.3 - Any duly authorized
representative of the Board may enter and inspect any property to
ascertain the state of compliance with the law and rules and
regulations of the Board.

Title 82 O.S.   936.  A Board has hearing on charges of violations
and authority to issue orders on notice.

Title 82 O.S.   937.  Penalties and Injunctions.

Violation of an order - $500.00 fine per day first ten continuous
days and $1,000 for each day thereafter or imprisonment for a term
of not more than 90 days or both such fine and imprisonment.  Each
and every day the violation occurs will constitute a separate
violation.

The offender shall be liable to pay the State an amount equal to
the sum of money reasonably necessary to restock such waters or
replinish such wildlife and all costs incurred in investigating,
locating or establishing the responsible person, firm or corporation
as determined by the Oklahoma Wildlife Conservation Commission and
approved by the Board.  Such amount recovered by the Board on behalf
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of the State in a civil action brought in the District Court shall
be divided among the agencies in accordance with expenses incurred
as determined by the Board.

It shall be the duty of the' Attorney General to take any action,
including an action for injunction.

This Act does not obviate other statutory provisions such as criminal
provisions for pollution of land.

The State of Oklahoma Ex Rel Pollution Coordinating Control Board v.
Kerr McGee.

     A.  Measure of Damages.

     B.  Punitive Damages.

     C.  Injunction.

A task force has been formed from this Act composed of individuals
from each department with individual duties to perform in the case
of a fish kill alert.  For example, the chemists from the Health
Department are to analyze water samples, the game rangers from the
Department of Wildlife are to count the number of fish, and the
other duties are separated out as will be noted by the head of the
Task Force, Glen Sullivan, in a later speech at the Seminar.

From the lawyer's standpoint, important evidence that we want in
addition to the chemical analysis of the water and fish, is an
area inspection, including any possible back water areas where fish
may have been effected, aerial photos and photographic maps, both
aerial and otherwise.  We want statements from the inhabitants of
the area and employees of the possible pollution source.  We need
weather reports from the area and temperature reports from the
stream or lake.  We want samples taken from the riverbed or the
banks of the lakes.  In any event, we want all of the evidence
prepared as if we were going to court on every case.

In the Cimarron River Fish Kill Case we are attempting to sue for
punitive damages, although it is not provided for in the statute,
on the theory that gross negligence of the polluters was as such
as to amount to an intentional act; therefore, they are liable for
punitive damages.  Otherwise, we are following the statute which
allows us to sue for the cost of replacement of the fish.

The end result of these lawsuits are not for political magnification
or for monetary gain, but to insure cooperation between city, state
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and federal agencies and private industry to protect our environment.
The real reason, and the only reason for enforcing pollution laws is
to ensure that our environment will be protected and that eventually
through the cooperation of all of us, its existence will be insured
for generations to come.
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           STEPS IN ESTABLISHING GOOD PUBLIC RELATIONS
                  WITH THE MEDIA AND THE PUBLIC

                               by

                          Diane Norvell

        School of Law, Univ. of Oklahoma, Norman, Oklahoma*
Communication is not one-sided.  In order to be a complete model,
it must be viewed from both ends.  And when acting in the capacity
of a state or public agency, it is most important that the agency
sees how its publics view its efforts as well as how the agency
sees itself.

How does John Q. Public measure the accomplishments?  Does he visit
with the various agencies to see its facilities, staff and latest
programs?  Does he peruse the latest annual reports to bring him up
to date?  Likely not, as much as we hate to admit.  Instead, he
usually gets a capsule of news from reading the headlines and first
paragraphs of his morning newspapers, or hearing the spot news on
the radio, or watching the 10 o'clock news before retiring to bed.
Consequently, media does play a strong role in influencing Mr. John
Q. Even the politicians, our representatives of federal, state and
local governments admitted its strength by spending millions of
dollars in the last general election in advertising to win Mr. Public.

With the population explosion comes the fight for brotherhood, popu-
lation zero, and last but not least pollution.  These are all examples
of man's need to be socially responsible.  Man wants to breathe clean
air, drink clean water and most of all enjoy recreational activities.
An outgrowth of these needs comes regulations pertaining to restrictions
on air emissions, solid waste, and discharges of waste into the streams.

Since Mr. Public is not a professional environmentalist but is concerned,
he wants to point the legal finger at those people who take away his
wildlife.  He wants polluters to pay for causing fish kills.

And what is your role in reporting the latest results of fish kills?
As I see it, your responsibility is:

     1.  Reporting the fish kill.

     2.  Reporting it in such a manner as he will understand.
*Former reporter with the Oklahoma Environmental Reporter, Oklahoma
City, Oklahoma.
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     3.  Letting him know in some situations there is no criminal
         but Mother Nature herself, as in the case of the Oklahoma
         droughts.

     4.  In the process, let Mr. Public know what you are doing
         about preventing and abating fish kills.

This is also a good time to put in a plug about your facilities and
if you need a more sophisticated lab or investigation tools let him
know.  It would mean more dollars and cents when appropriation
considerations come before Congress and the state legislature.

Here is an outline in establishing good public relations:

     I.  ORGANIZATION OF PUBLIC INFORMATION SOURCES

         A.  Set up a central public information system.

             1.  Name a public information officer (PIO) to organize
                 and write all press releases for your agency.

             2.  In the case of fish kills and other incidents which
                 require the efforts of several state agencies, an
                 expedient way of releasing information is to appoint
                 one PIO to funnel information to the press.  This
                 system would save overlapping stories and man hours
                 to boot.

             3.  If the coordinated system is adopted, make sure
                 participating agencies refer the press corps to a
                 particular spokesman who will speak for the group.

             4.  If the coordinated system is not used, only release
                 information on your particular aspect of the fish
                 kill investigation and try to keep from making
                 comments to the press which would cast a shadow
                 over the other agencies.

             5.  By all means don't keep your PIO man out in the cold.
                 Keep him up-to-date on the latest investigation
                 results and give him guidelines on what information
                 he is to release.  An old story is not NEWS.

             6.  Keep a file on all fish kill stories.  This
                 collection will aid your agency in handling other
                 fish kills.  You can use the file for composing
                 a thorough annual report.
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             7.  Give your PIO man feedback.  Criticize his stories
                 and give him ideas for writing better releases.
                 Don't forget to compliment him when he writes
                 'Pulitizer Prize Winning' articles.


         B.  Set up a working relationship with the press corps.


             1.  Know the reporters who cover conservation activities,
                 for they write stories read by a large number of
                 subscribers.


             2.  Abide by newspaper deadlines so your story will be
                 timely.


             3.  Don't make the article too technical.  If the
                 reporter doesn't understand the concept, neither
                 will the reader.


     II.  LEGAL LIMITS


         Premature fish kill investigation reports to press may
         harm the court suit.  Careful analysis should be made
         when releasing information on the following:


         A.  Fish kill losses.  Don't get locked in with a specific
             number.


         B.  Give possible pollution sources but be leary in naming
             alleged polluters until formal charges are brought.


As you can see, communication is being able to relate your ideas
effectively and in such a manner that those people who receive
transmission will understand your thoughts.
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       ALABAMA PROGRAM FOR THE INVESTIGATION OF FISH KILLS


                               by


                         Sam L. Spencer
         Chief, Fisheries Section, Game and Fish Division
                 Alabama Dept. of Conservation



In Alabama, we have a two phase program for the investigation of
fish kills.  This consists of the Alabama Water Improvement Commission
(AWIC) and the Game and Fish Division of the Alabama Department of
Conservation.


                 AUTHORITY TO CONTROL POLLUTION


The authority to investigate and control water pollution is vested
primarily in the AWIC; however, the Game and Fish Division has been
designated to assist, and damages collected are to be used by Game
and Fish for "improving fishing in the affected area".


The AWIC has a central office in Montgomery and a Mobile lab for
extended studies.  At present they have no field offices.  The
Game and Fish Division has approximately 130 field personnel and
are better situated to investigate fish kills.  We have 105 Conser-
vation Officers, 12 field Game Biologists and 13 field Fisheries
Biologists.  We have two-way radio contact with these field personnel
through a state-wide radio network.  In addition, an airplane is
usually available when needed.  The fisheries biologists have the
primary responsibility for investigating the fish kills, however,
other field personnel are available if needed.


                       REPORT OF FISH KILL


In Alabama fish kills are normally reported in one of four ways.


     1.  Reported by citizens to Game and Fish personnel.


     2.  Reported by citizens to AWIC personnel.


     3.  Reported by industries and municipalities to AWIC.


     4.  Observed by field personnel.
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We have plans for a leased telephone line so individuals can report
fish kills at any time of day or night.  Upon receipt of notice of a
fish kill, the informants name, telephone number and location of kill
is reported to the AWIC, the Alabama Department of Conservation
officials and to the district fisheries biologist nearest the kill.
Fish kills are our highest priority item and every effort is made to
reach the kill as soon after it is reported as possible.

                  INVESTIGATION OF FISH KILLS

Ideally a fish kill is investigated by both the AWIC and the Game
and Fish Division.  The AWIC technical personnel would collect
water and fish samples, conduct basic water chemistry and attempt
to determine the cause of the kill.  The fisheries biologist would
determine number, size, species and location of dead fish or other
aquatic organisms.  They would check for possible causes unrelated
to pollution such as disease and parasite problems.  When this work
is completed, they would assist AWIC personnel with their investigation.
Assistance is often rendered by other agencies such as the Tennessee
Valley Authority, and the Environmental Protection Agency.  The
Environmental Protection Agency has been most helpful in analyzing
samples collected during fish kills.  Often the Game and Fish
personnel is the only person on the kill and it falls his lot to
conduct the entire investigation.

                             REPORTS

After the kill, a written report is prepared by the fisheries
biologists and the AWIC personnel that conducted the investigations.
A copy of each report is submitted to the State Attorney General's
office for review.  The reports are reviewed at regular meetings
of the AWIC board.

                            LAW SUITS

Potential law suits are usually referred to the Attorney General
for decision and legal action.  Such suits are handled by the State
Attorney General's office.  Under a new law the Attorney General
has the power to independently enforce provisions of the pollution
law if the AWIC fails to.  Suits may be filed for compensatory
damages for the value of the fish in cases where negligence is
suspected.  Punitive  damages may be pursued in cases of willful
or wanton destruction of fish.  This is almost impossible to prove.
Under our new law the penalty for violation of water pollution laws
is from $100 to $10,000 and each act or day is a separate offense.
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Another significant change in our new law is the provision that "If
the pollution has caused damage to fish and/or other wildlife in
excess of $5,000, as determined by the Department of Conservation,
the damage shall be presumed to have been the direct and proximate
result of negligence of the person shown to be responsible for
such pollution and the burden shall then be upon such person to
prove freedom of negligence in causing the pollution in such cases."


                  THE AWIC - PAST AND PRESENT


In the last session of our State Legislature the make-up of the AWIC
was drastically changed.  The old commission was originally set up by
law in 1949, changed slightly in 1953 and again in 1965.  The commission
was composed of 14 members as follows:


     1.  Chairman - State Health Officer


     2.  Vice Chairman - Director of Conservation


     3.  Commissioner of Agriculture and Industries


     4.  State Geologist


     5.  One representative of municipal government


     6.  One representative of county government


 7 & 8.  Two representatives of wildlife conservation


     9.  One representative of the mining industry


    10.  One representative of the textile industry


    11.  One representative of the chemical industry


    12.  One representative of the lumber industry


    13.  One representative of the paper industry


    14.  One representative of the metals industry


It has often been stated that in Alabama we had the "foxes guarding
the hen house."


The commission also included a technical staff of merit system
employees composed mainly of engineers, chemists, and aquatic biologists.
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This commission was abolished effective October 1, 1971, by our
State Legislature.  The members of the new commission are as listed
below:


Non-Voting Members:


     1.  Chairman - State Health Officer


     2.  Vice Chairman - Director of Conservation


Voting Members:


     1.  Physician qualified in water-borne diseases


     2.  Professional engineer qualified in water resource
         management and water supply


     3.  Attorney qualified in fields of water supply and
         riparian rights.


     4.  Two residents of the State for two years with no
         specialized experience required.


It was further specified by law that the members of the commission
could be from industry but not from industries that discharge effluent
into the public waters of the State.  The voting members serve four
year staggered terms, are chosen by the Governor and serve at his
pleasure.  He has the power to replace any member at any time.


                    PRESENT NEEDS IN ALABAMA


We feel we have a good setup for investigating fish kills in Alabama.
The main items in my opinion where improvements are desired are as
follows:


     1.  The appointment of commission members that favor clean water.
         At this time, the members have not been appointed yet.


     2.  The AWIC needs additional funds to establish district offices.
         This would improve detection and investigation of the kills.


     3.  A few more successful law suits may be helpful in reminding
         these few industries that are slow to clean up that the
         State of Alabama plans to protect its aquatic resources.
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            ARKANSAS FISH KILL INVESTIGATION PROGRAM

                               by

                           Neil Woomer

         Arkansas Dept. of Pollution Control and Ecology
Officially, the responsibility and authority for investigating
pollution caused fish kills in Arkansas rests entirely with the
Arkansas Department of Pollution Control and Ecology.  As a
practical matter, such investigations are carried out with the aid
of the Arkansas Game and Fish Commission, particularly in the areas
of reporting of fish kills and making damage evaluations.

Referring to the attached flow diagram, fish kill investigations are
carried out in the following way.  Beginning with the initial observer,
the fish kill is nearly always reported to either a Game and Fish
Enforcement Officer, of which there are 125 spread throughout the
State; to the Game and Fish Commission Fisheries Division, which has
12 District Fisheries Biologists; or directly to the Department of
Pollution Control and Ecology, which has 12 Field Inspectors throughout
the State and 6 Ecologists in the Central Office at Little Rock.  If
an Enforcement Officer receives the report, he radios the Fisheries
Division which then contacts Pollution Control.  If Pollution Control
gets the report first, the Fisheries Division is immediately notified.

Once the report has reached the ADPC & E Ecology Section, an attempt
is made to contact the nearest District Field Inspector.  If this
can be done and he can reach the site of the kill more quickly than
someone from Little Rock, he makes a preliminary investigation,
gathering pertinent information and taking the all-important first
set of samples.  If no Inspector is available a Biologist is immediately
dispatched from Little Rock.  All fish kill reports are investigated.
If the preliminary investigation shows that the fish kill is substantial,
the Inspector immediately calls Little Rock and a team of biologists go
to the scene to do a complete work-up.  After the field investigation
is completed, all water samples are turned over to the ADPC & E Chemistry
Section, which is capable of doing all necessary analyses, and all
biological samples are analyzed by the Ecology Section.  Meanwhile,
the Arkansas Game & Fish Commission Fisheries Biologists make a
complete damage evaluation which is submitted to the investigating
Pollution Control Biologist who then writes a complete final report
that is turned over to the legal and administrative people who then
proceed with appropriate legal action.
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Violation of the Arkansas Pollution Control Law is a misdemeanor
and is punishable by a maximum fine of $250.00 or up to one year
in prison.  Arkansas has no law enabling the State to recover damages
caused to its natural resources by pollution, although the industries
involved in several fish kills have more or less voluntarily restocked
fish equal in value to the fish killed by their discharges.  Such a
law is very badly needed in Arkansas.


The only other major problem that lessens the effectiveness of the
Arkansas program is that of communication.  We have no radios and
the only way Field Inspectors can be contacted is when they are
in their offices, which they seldom are.  We need radios for the
Inspectors and for the Ecology Section vechicles to tie in with
the Arkansas Game and Fish Commission radio network which is Statewide,
If this can be done and an adequate damage law passed, Arkansas will
have an ideal fish kill investigation program.
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ADPC&E <-
 1   L_
             ARKANSAS FISH KILL  INVESTIGATIO
                        INITIAL
      AGgFC ns
-> HIV,  (\? '-Inl
FIFLD INSPECTOR (?)
  (If Closer)
      ^preliminary
        Fr.nL.nnY SFCTP"!
                               V
ADPCRE
CHEMISTRY SECTION
      •WATFR, rlJD, TISSHF  SAMPLE
AGgFC
FISHERIES
               Cr.izes , I'uinborr-. , ,"no" l^.-, ,  , n 1 no)
                                      "I
                               30
WC f N
 (1 PS Wardenr,)
                                                    ,  Plankton,

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             INVESTIGATION OF FISH KILLS IN MISSOURI

                               by

                           Frank Ryck

               Missouri Department of Conservation
The Missouri Department of Conservation is the Wildlife regulatory
agency and the Missouri Water Pollution Board is the agency having
legal responsibility for pollution control.  Within the Conservation
Department, the Water Quality Branch of the Fisheries Research Section
is most directly involved in fish kill investigations.  At the present
time, the Water Quality Branch is composed of a Chemist - Supervisor
and two Fisheries Biologists.

There are basically 5 sources of original information of fish kills
that occur in Missouri.  Most fish kills are reported to the Water
Quality Branch by Conservation Agents.  The Conservation Department
has 128 Agents - one in each county.  The Agent has an intimate
knowledge of his county and is in close personal contact with concerned,
conservation-minded citizens.  These citizens provide the Agent with
most of the initial reports of fish kills.  The Department of Conser-
vation has 8 Fisheries Management Biologists whose duties bring them
in close contact with the Conservation Agents and concerned citizens.
We frequently receive notification of fish kills from the Management
Biologists.  The Water Pollution Board sometimes receives the initial
report of fish kills from industries, municipalities, or other state
agencies.  In a number of fish kills, the first report has come from
direct observations by members of the Fisheries Research Section.

When notification of a fish kill is received by the Water Pollution
Board or the Conservation Department the other agency is immediately
notified.  The Water Pollution Board notifies the Environmental
Protection Agency, the Missouri Division of Health, and any other
agencies that may be concerned.  In cases of very large fish kills
the Conservation Department also notifies the Environmental Protection
Agency.  After this process of notification a joint investigation is
initiated by the Conservation Department and the Water Pollution Board.

A typical fish kill investigation begins with the notification of
Water Quality personnel.  The person reporting the fish kill is
expected to have roughly determined the distance of stream affected,
the seriousness of the kill, and possible sources of the pollution.
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He is also expected to have collected water samples above, in, and
below the kill zone.  If possible, a sample of the pollutant is
collected.  Water Quality and Water Pollution Board personnel
proceed to the kill site and meet with the reporting personnel.
Coordination of movement is facilitated through the use of radio-
equipped cars and airplanes.  County files are consulted before
any investigation is initiated to help pin-point possible sources
of pollutants.

Once Water Pollution Board and Water Quality personnel are at the
scene of the kill a full scale investigation is begun.  Additional
water samples are collected, and field analyses are run to determine
the following parameters:  temperature, pH, NH3, dissolved oxygen,
turbidity, acidity or alkalinity, conductivity.  Concurrently, sample
counts of dead fish are made, and estimates of the number of fish
killed are based on these counts.  The size and species of dead fish
in the counts are established and monetary damage estimates are
calculated by consulting the publication of the Pollution Committee
of the Southern Division of American Fishery Society relating to
Monetary Values of Fishes.  Whenever possible, post-kill benthos
and fish population studies are made to determine the types and
numbers of organisms that survive.  Water samples are transported
to the laboratory where Water Quality- personnel conduct additional
analysis, and conduct bioassays.  Samples suspected of containing
heavy metals, or pesticides are turned over to the Water Pollution
Board or the Fish Pesticide Laboratory for analysis.  When the
investigation is completed a report is written.  Copies are filed
with the Water Pollution Board, Conservation Agents, and the
Attorney General's office.  When possible, criminal charges are
filed with the county Prosecuting Attorney against the polluter
for violation of the Wildlife Code of Missouri.  The Attorney
General's office is routinely filing for damages to the State
whenever fish are killed.
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              NON-TOXICANT RELATED FISH MORTALITIES

                      Introductory Remarks

                               by

                    Dr. Robert C. Summerfelt

            Leader, Oklahoma Cooperative Fishery Unit
         Oklahoma State University, Stillwater, Oklahoma
This seminar is designed to serve as a forum for dissemination of
knowledge and for exchanging views on the complex problem of fish
kills for State and Federal agencies responsible for protecting our
health, environment, and fish and wildlife resources.  The focus of
the conference is fish centered, however, not because most agency
representatives attending the conference have the responsibilities
to protect fisheries as valuable natural resources, but, because
dead and moribund fishes are conspicuous indicators (barometers)
of the state of an environment which needs monitoring for our
survival, livelihood and general well-being.

We have heard it mentioned many times that a fish kill is a conspicuous
red flag but the real value of this signal is often overlooked even
by communities with intakes on major rivers susceptible to environmental
contaminants.  Keeping an eye out for a fish kill on a water supply is
vital to the community because our rivers are arteries of a vital
life support system for which dead and dying fish serve as an early
warning system.  Investigations of fish kills should be as vital to
a municipality as caged canaries were to coal miners.  The latter
carried canaries into labyrinths of the subterranena tunnels they
worked, and, so long as the canaries were alert, the men did not fear
asphyxiation for the dreaded coal gas because they knew the birds
would collapse from the coal gas before the concentration became fatal.

Our conference deals head-on with the causes of fish kills but few
attending this seminar subscribe to a view that the absence of
conspicuous fish kills Is an indication of a healthy environment.
An analogy will clarify the point:

     "If a tree falls in the forest but unseen by man, does it
make a sound?"  No!  By definition, sound is a vibration that
stimulates the auditory nerves and produces sensations of hearing.
If man is not there to hear, sound is not produced, and vis-a-vis
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the environment, an absence of a fish kill means losses have not
occurred and all is well.  This line of reasoning mimics the expression,
"What you don't know won't hurt you."

In looking only at fish kills we are looking at only the part of the
iceberg which is above water.  More radical changes may be associated
with gradual attrition and changes in species composition of communities
than the changes associated with a fish kill.

It seems that more profound environmental factors affecting fish
populations operate in a slow or not easily perceived manner.

Evolution, for example is a process of extinction and speciation
resulting from gradual changes and replacement.  The most well-known
example of the effect of human perturbation on evolution is the case
of industrial melanism in the "peppered moth" (Biston betularia)
which occurs in light (peppered) and dark (melanic) forms.  In 1848
the dark type was very rare near Manchester, England, but in years
thereafter, the relative frequency of the dark form was observed to
increase until today, only the dark forms are found near Manchester.
Prior to industrialization, lichens covering the trees give the surface
a variegated surface against which the lightly peppered moth was hard
to see and the dark form quite conspicuous, but with industrialization
based on consumption of fossil fuels, tree trunks blackened by soot,
made the light moth very conspicuous but concealed the dark moth.
Industrialization resulted in a change in gene frequencies in the
population because it reversed natural selection (H. B. D. Kettlewell).
Carried far enough, differences between the first and last series of
hereditarily related organisms can lead to speciation, but more
importantly, it leads to changes in species composition in a population
because just as the survival of the dark form was competitively better
than the white under the altered conditions, changes in the environment
can favor other species, species which are more resistant.

In natural lakes and reservoirs, gradual eutrophication favors carp,
buffalo, gar and bullheads and a concomittant reduction or elimination
of bass, bluegill, walleye, and other desirable species.  Survival
and development of pesticide resistance often is associated with a
partial loss of other elements of physiological vigor, or less fit
for natural stress.

My next examples relate to problems of reproduction, although it is
obvious that every species requires successful reproduction and
survival of at least two young from every set of parents, reproductive
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failures resulting from environmental conditions are often overlooked
as important problems.  Poor recruitment of largemouth bass in a
certain Oklahoma reservoir, barely enough to sustain the population,
has been found to be due to wind induced turbulence and lack of
suitably protected spawning sites.  Poor recruitment of largemouth
and smallmouth bass in a certain Oklahoma river has occurred because
releases from a hydroelectric plant, with penstocks located in the
hypolimnion, maintains a water temperature too low to allow spawning.
Fish have a sufficient high fecundity and have evolved with natural
stresses and survived natural catastrophes, but persistent, sublethal
effects can change and destroy natural ecosystems and fisheries
resources with far greater effect than the natural stress.

The examples are those of insiduous environmental induced changes
caused by man's perturbation of natural ecosystems.  No fish kills
need to be involved, yet the effects are very pronounced.  I know
that you are aware of similar cases of environmental alterations which
have not produced fish kills, but need critical assessment.  Legal
liability for insiduous changes, although as I believe are of greater
magnitude than the fish kill, are not adequately covered by the
existing administration-legal framework which operates to assess
losses related to costs of dead fish observed in massive kills.

Small or reoccurring kills are often neglected, receive only a cursory
investigation, and the concerned agencies are by necessity forced to
concentrate on the major kills.  Habitat degradation is rarely
investigated and there are few legal precedents for compensatory
charges for habitat alteration.  Although it is a violation to throw
or allow to run into waters any substance injurious to fish, the
laws usually provide penalties based on the number of fish killed.
In some cases, it allows a penalty for a condition which would kill
fish even where no fish are killed, however, this still doesn't
apply to the situation of habitat degeneration.

Regarding penalties and compensatory payments for damages it may be
biologically unsound to replace the fish except that these costs
are symbolic or representative of the environmental degredation.

               NATURAL VS MAN-INDUCED MORTALITIES

Fish mortalities result from a variety of causes, some of natural
origin and some man-induced, a prompt, well organized investigation
is necessary in order to provide the information needed to determine
the cause of the fish kill and the source of the killing agent as
well as to alleviate the possibility of reoccurrence.  Fish kills
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as used in this conference refers to sudden mortalities, not chronic
losses, and mortalities of sufficient magnitude because most small
fish kills are inadequately investigated.  Moreover, fish kills in
this conference refers to the mortalities, large or small, due to
natural or man-induced.  Unfortunately, some investigators make a
distinction between epizootics and other "natural" mortalities as
fish "die-offs" , and mortalities which are man-induced as "fish-kills",
This leads to a rather confusing situation in an investigation where
occurrence of dying fish cannot be called a fish kill unless you know
it is man-induced, and you do not call it a fish-die-off unless you
know it is attributable to natural causes.  I personally favor use
of fish kills as a general term for both types.
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             NATURAL VARIATIONS IN FISH POPULATIONS

                               by

                        Austin K. Andrews

             Bureau of Sport Fisheries and Wildlife
                Oklahoma Cooperative Fishery Unit
         Oklahoma State University, Stillwater, Oklahoma
The intent of this discussion is basically a plea for recognition
of the need for in-depth study of our fish stocks before obvious
fish kills occur and especially before occurrence of those insidious
changes that alter species and stock composition just as inexorably
and possibly more permanently than the spectacular kill.

To address the question of natural variations in fish population
size without being merely anaectdotal is a real challenge because
although we have an extensive literature concerning the timing, extent
and consequences of these fluctuations, we know very little of the
mechanisms involved with causality.

The fact that fish population size and structure varies without the
interference of man has intrigued and excited fishery biologists from
the time of the first investigations, because variation indicates
controlling factors that are not static and intimates that once these
factors have been identified, they may be modified and in so doing
allow us to manipulate or "manage" the resource.  Therefore, let us
examine our approaches to identifying and dealing with these fluctu-
ations and how our knowledge may then be applied to accessing
responsibility for these variations.

            THE FISHERY BIOLOGISTS' VIEW OF MORTALITY

Fishery biologists are primarily concerned with making fishermen happy.
Now the surest way to have happy fishermen is to be sure that they are
able to catch a reasonable number of fish of the right size and species
on each fishing trip.  Therefore, in order to achieve this goal (i.e.,
happy fishermen) our fishery biologist will attempt to manipulate the
various fish stocks for which he is responsible.  When he performs
these manipulations he must have some way of knowing whether he is
adjusting his stocks in the right direction and perhaps even more
important, he must be able to predict the stock composition several
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years in advance.  To determine the current status of the fish
populations (with respect to its contribution to the fishery), he
can sample fisherman catch; however, to predict, he must look directly
at the fish stocks and the population dynamics that will determine the
relative size of any population when it first begins to contribute to
the catch.  Therefore, the essence of management of fish stocks is the
study of population dynamics and population dynamics consists of the
balance between natality, growth and mortality.  Mortality is often
studied because it is relatively easy to estimate and allows us to
examine stock composition under the basic assumption that the decline
in stock density from year to year is due to mortality and the ratio
of stock densities can thus be used to estimate mortality (Gushing, 1968),

                      SOURCES OF MORTALITY

This brings us to the crux of the whole matter:  when a fishery biologist
looks at mortality he traditionally (and almost automatically) divides
it into 1) fishing mortality and 2) natural mortality (unusually gotten
by difference and includes all fish losses not attributable to fishing).
Now our category of natural mortality contains a very mixed-bag of
sources of death, and to reach a definition of what we will consider
as "natural" and "unnatural" mortality that would be satisfactory to us
all becomes an exercise in semantics.  For instance, if we can consider
as "natural" only those systems unaltered by man, then we're fresh out—
but serious attention must be directed to how much of man's casual
influence we can accept before he is the unintentional controlling
factor in the aquatic ecosystem.

As I mentioned earlier, fish populations did and do vary, sometimes
drastically, without the aid of man.  We give tacit acknowledgement
to this by the very title we give to these studies of ... population
dynamics.   What, then, are same of the factors that contribute to these
natural variations?  Alphabetically we can quickly list:

     Behavior;
     Catastrophies in the environment, both immediate and evolutionary
          in scope;
     Chemical, again our attention is usually directed to seasonal
          or yearly fluctuations, but geologic time references are
          applicable;
     Critical life stages, although not a source of mortality in
          itself, moving from one to the next is a singularly precarious
          time in the life of a fish and one should examine these
          tranistional periods to determine if a disproportion amount
          of mortality is occurring (Kramer, 1969, has assembled a
          nice bibliography dealing with early mortality in freshwater
          fish) ;
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     Disease, Dr. Meyer will discuss the role of diseases in fish kills
          later in this seminar;
     Food, or really the lack of the right kind or size at the right
          time;
     Genetic;
     Light;
     Parasites, I believe Dr. Meyer will also touch upon this subject;
     Predation, even discounting man this is reputed to be the source
          of the greatest amount of mortality in most fish stocks,
          however, with the possible exception of mortality occurring
          early in life, I will argue most vigorously against this
          as s ump t i on;
     Radiation;
     Senility, this actually accounts for very little of the total
          mortality suffered during the tenure of a given year class;
     Siltation;
     Water temperature;
     Water level fluctuations;
     Wind and wave action;
     and on and on.

The point to be made here is that fish (being ectothermic, relatively
stenothermic and, in the case of our freshwater fishes, trapped in a
very limited environment), are completely at the mercy of the vagaries
of nature.

                  BALANCE SHEETS AND MORTALITY

We still are no closer to accessing responsibility for our mortality
because although we are able to be quite precise in our estimates of
the magnitude and timing of fish deaths, we are still, in the vast
majority of cases, most uncertain as to the actual cause(s) of
mortality.

Regier and Robson in their chapter on Estimating Population Number
and Mortality Rates in Shelby D. Gerking's Freshwater Fish Production
point out that:

     "Close monitoring of the dynamics of a single freshwater
     fish population is often frustrating and discouraging
     because of the occurrence of violent and inexplicable
     fluctuations.  Pattern and regularity in freshwater fish
     production will emerge more clearly by combining observations
     made at different locations and years over a geographic
     region, as is done in the analysis of agricultural
     production."  pg 32.
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I don't really believe that it Is quite as black  a picture  as  they
paint.  Although it is true that these violent  fluctuations  do occur,
to write them off as inexplicable is only defeatism.  However,  the
charge is clear.  We must know the fish stocks  with which we are
dealing!

More often the variations that we may expect  are  much more  limited.
If we accept, a priori, the concept of carrying capacity being
representative of the productive capacity of  a  body of water,  then we
would expect only minor yearly fluctuations in  productive capacity and
consequently in total fish biomass.  What then  are the clues to change
within our system?  (The theory is present and  continually  being
refined.)  We must look to the dynamics of the  individual populations
and examine the structure and patterns of age and growth together with
species diversity indices to tell us of major shifts in system structure.
In this way, we can gain insight into not only  the timing of fluctuations
(our predictive element) but into casuality and control within the
system.  Once an understanding, of the types  and  magnitude  of  factors
driving and steering the system has been gained,  we will be  in a position
to sort and access the t-ffects and fill in our  balance sheet for
mortality.

                  AN OVERVIEW OF THE TECHNOLOGY

The way in which we describe community stability  is to determine
relative abundance or relative total biomass  of the population of all
the fish species in the system.  Since the relative abundance  of the
species is a condition of dynamic balance resulting from the way in
which the environments productivity has been  divided, then,  even if
total productivity remain-- constant a change  in one species' numbers
(even if only temporary) will cause a series  of oscillations throughout
the rest of the system.  Therefore, once the  structure of the  system
has been defined, change in one component or  species allows  us to
predict the changes, both in '•<.-spect. to time  and  magnitude,  in the
rest of the components '.specie^;.

We are able to use the same general procedure in  describing  the relative
stability of a single species.  Relative size of  the individual year
classes over time does tend to he constant.  Mayhew (1956)  found that
blue gills in an Iowa IJKI- exhibited a fairly  stable population over a
period of 14 years, and Mri-udden and Cooper (1962) stated that brown
trout recruitment La =-. i •< different populations  appeared stable with
only occasional variations caused by flood or drouth.  Nikolski (1969)
in his book Theory o_f Hsh Population ]tynajn:L_cs_  sums it all  up  stating
that each specic=s has i :.., own mortality rate with its own distribution
over age groups.

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This information now allows us to draw some generalizations about age
structure in our fish stocks.  First, we expect stability or equal
sizes of year classes over time.  Second, we expect variation that
does occur to happen early in life, usually the first year is when the
relative year class size is established.  Even though no year class
operates independently within a population, the magnitude of the variation
should be identifiable throughout the tenure of that age class (Figure 1).
Note in the figure that if we had made a point estimate of the mortality
characteristics in this population at year 3 or year 4, we might attribute
the apparent increase in mortality (from 0.5 to 0.75) to the wrong cause,
for example a logical explanation might be that this was the time that
this species first became vulnerable to fishing.  Extreme care must be
exercised in interpretation of data gathered from a population over
a short period of time.  Last, these characteristics of population
structure allow us to formulate a set of assumptions for support of
our theory of mortality estimation.  These are:

     1.  The age groups in question were equal in numbers at the time
         each was recruited to the fishery.  (Probably least often
         satisfied and the proximate cause of most natural population
         fluctuations.)

     2.  The mortality rate is uniform with age, over the range of
         age-groups in question.

     3.  Since mortality is composed of fishing and natural mortality,
         each of these, individually, is uniform.   (Some authors
         have seriously questioned whether this is ever satisfied.)

     4.  There is no change in the population mortality rate with time.

     5.  Samples are taken randomly from the age groups involved.

Of course, in practice you never expect to completely satisfy these
assumptions.  However, we do have some idea of the effects of failure
to satisfy them on our estimates.  The outgrowth of this is that while
fluctuation in individual year classes is expected, we don't expect
general depression of all year classes within a short time span.  If we
do observe this latter type of population change,  we should be able to
identify a climatic catastropby or begin to look for the possibility
of man's influence.

Another point to bear in mind is that natural mortality is highly
seasonal.  We expect loss in a new year class to occur predominately
within the first few months of life.   Older fish will usually succumb,
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 •               Figure  1.  A classical, hypothetical example of the effects, over
                           time,  of  a poor year  class.
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              = 0.5
NORMAL
              POOR  SPAWN
                         YEAR 2
                        YEAR 4
YEAR  5
              YEAR 6
li  li!   (V
V  VI   0
  AGE CLASS
                A3

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providing the major losses are not due to predation, during periods
of stress which are usually associated with either reproduction or
the period when the aquatic environment reaches the farthest point
from optimum levels.  This usually may be referenced by temperature;
therfore, the critical period for a warmwater fish such as a bluegill
will probably occur during the latter part of the winter while the
critical period for one of tinj trouts may occur during mid-summer.

                           DISCUSSION

In most freshwater systems (and I say most as a broad generalization
that certainly has many exceptions) the external or physical variables
exerting the greatest effect on a system are temperature and water
level both operating within a critical time vector.  According to
most analyses in the literature concerned with the question on natural
variation, if we add the internal control of predation (and I would
rather substitute disease and parasitism), especially in the first
few weeks or months of life, we have identified those factors most
responsible not only for the ultimate size and structure of a population,
but also for the fluctuation and variation that we observe.

So let us turn from consideration of those few freshwater species that
are typically cyclical with impressive displays of "boom and bust" that
have intrigued us and stolen much of our investigative energies and
examine the subtleties of the much more common, and in the overview,
more important minor fluctuations and "creeping" change that indicates
overall system stability or direction of system movement.

                           CONCLUSION

So, while I urge you to continue with the important monitoring programs,
that have been described earlier in the seminar, as a viable means of
preventing future catostrophic losses due to man's activities, I would
also urge you to look also to the potentially more dangerous problem
(in the long run) of insiduous changes that are not normal geologic
and evolutionary change.  These are the ones that will be responsible
for total change and in some cases total loss of the recreational/
commercial fishery resource.

In conclusion, since we, upon entry into the realm of scientific inquiry
have, at least tacitly, acknowledged and accepted a deterministic
philosophy, let us get about the business of sorting out the effects
and identifying the proper causes without losing awareness of the role
of proximate and ultimate causality.  Then and only then may we assign
responsibLlity to man for his actions and address ourselves to the
questions of direction and type of control within the aquatic ecosystem
that he may be allowed to exercise.

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                        LITERATURE CITED



Gushing, D. H.  1968.  Fisheries Biology, A Study in Population Dynamics.
     The University of Wisconsin Press, 200 pg.


Kramer, R. H.  1969.  A preliminary bibliography on extent and causes
     of early mortality in freshwater fish (diadromous fishes excluded).
     F.A.O. Fish. Cir. #307, 20 pg.


Nikolskii, G. V.  1965.  Theory of Fish Population Dynamics as the

     Biological Background for Rational Exploitation and Management
     of Fishery Resources.  Oliver and Boyd Ltd, Edinburgh, 323 pg.


Regier, H. A. and D. S. Robson.  1967.  Estimating population number
     and mortality rates, pg. 31-66.   In The Biological Basis of
     Freshwater Fish Production, edited by S. D. Gerking.  Blackwell
     Sci.  Pub., Oxford, 495 pg.
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               THE ROLE OF DISEASES IN FISH KILLS

                               by

                          Fred P. Meyer

             Bureau of Sport Fisheries and Wildlife
                Fish Farming Experimental Station
                       Stuttgart, Arkansas


                          INTRODUCTION

Mortality due to natural causes is probably the largest single
cause of death in a fish population.  It has been shown that unless
killed by some introduced factor (usually due to man) or predation,
the most likely cause of death of fish will be old-age (Ricker, 1954).
Although many fish may die of disease during a year, seldom does
the problem develop to epizootic proportions.  Epizootics in nature
caused by disease are often induced by a deterioration of the
environment in which the fish live (Reichenbach-Klinke and Elkan,
1965; Petrushevski and Shulman, 1959).  In this respect, it might
be noted that the same situation exists in the managed water areas
on commercial fish farms (Meyer, 1970).  Stress plays an important
role in the initiation of disease and may often be overlooked unless
the observer is alerted to the possible effects of low oxygen,
sudden temperature changes, or pollution of any kind (Wedemeyer,
1970).   It should also be noted that many of the organisms which
cause serious problems on fish hatcheries and fish farms are present
in low numbers in natural habitats.

The host-parasite relationship between the fish and their parasites
has developed to the point where the parasites or pathogens should
feed only upon surplus tissues and energy of the host.  Successful
parasitism requires that the parasite species does not endanger the
survival of the host.  Should the parasites feed excessively upon a
host, either because of excessive numbers or due to a reduced vitality
of the host brought about by other factors, the net result is death
of the host, a loss of habitat for the parasitic organisms, and a
subsequent reduction in the population of parasites (Chandler, 1955).
If, on the other hand, host populations have developed proportions
beyond their normal food supply and beyond the carrying capacity of
the habitat, their resistance may be sufficiently reduced that
disease will remove the excess and help bring the host population
back in balance with its environment.  Thus, under normal circumstances
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of adequate food and favorable environmental conditions, disease
will seldom be a major factor in the elimination of a species
since the host-parasite relationship is self-limiting on both the
host and the parasitic organism.  Epizootics must then occur only
whenever some factor has entered the picture to disrupt the balance
(Baer, 1952) and observers must be alert to identify the causative
agent, be it over-population, starvation, pollution, or other
environmental change.

              RECORDS OF DISEASE-INDUCED FISH KILLS

While the foregoing emphasizes the infrequent occurrence of
epizootics in natural habitats, records are available describing
situations in which it was determined that disease was indeed the
primary factor.  It is also likely that disease plays a significant
role in many other losses but that it has been overlooked.  The
following list indicates that nearly all major groups of parasitic
organisms have the potential of initiating epizootics in natural
waters.  In addition, readers should keep in mind that those
organisms which cause epizootics on hatcheries and fish farms have
the potential of doing so in lakes and streams.

Fish kills due to bacteria are frequently overlooked since few
observers are trained to recognize symptoms of bacterial disease
and even fewer have the specialized facilities at hand to make the
necessary isolations and cultures needed to verify the cause.
Records at the Fish Farming Experimental Station Diagnostic Laboratory,
Stuttgart, Arkansas contain numerous records of fish kills in natural
waters caused by bacteria.  Lakes with high populations of stunted
black bullheads or crappies are frequently hit with epizootics of
myxobacterial infections (Columnaris disease).  A case of particular
interest concerns a reported loss of largemouth bass in Lake Maumelle
(Arkansas) in which only bass over 2 pounds were dying, with a
preponderance of dead fish over 4 pounds.  Field observations
revealed that a high number of stunted, emaciated, 6" gizzard shad
present in the lake were also dying.  These had gone unnoticed by
the sportsmen whose only concern was that the large game fish were
lost.  Gizzard shad in varying stages of dying were evident over
much of the lake.  Large bass were feeding actively on the moribund
shad.  A study of both species indicated a heavy infection of
Columnaris disease in the shad which was, in turn, transmitted to
the bass when they fed on the distressed fish.

Pacha and Ordal (1970) report that numerous outbreaks of Columnaris
disease have occurred in natural and hatchery populations since 1944
in both warmwater and coldwater environments.  Epizootics have been
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reported in Atlantic salmon (Elson, 1968; Pyefinch and Elson, 1967);
Sockeye salmon (Colgrove and Wood, 1966); stonerollers (Lennon and
Parker, 1960) and among "coarse fish"  (Bottomley and Holland, 1966).

Snieszko (1958) reports that high water temperatures reduce the
resistance of fish to Columnaris disease and Pippy and Hare (1946)
indicated that copper and zinc pollution induced conditions which
rendered the fish susceptible to Aeromonas liquefaciens.

The incidence of bacterial hemorrhagic septicemia caused by Aeromonas
liquefaciens has shown a stress-related pattern similar to that of
Columnaris disease.  On fish farms, outbreaks are a common occurrence
and are closely correlated to stresses of low oxygen, high temperature,
and handling.  Epizootics in natural waters are also common.  Major
epizootics have been reported from a wide variety of warmwater fishes
(Haley, Davis and Hyde, 1967; Reed and Toner, 1942; Rock and Nelson,
1965; Wagner and Perkins, 1952; and Meyer, 1964).  Rock and Nelson
noted that low levels of oxygen were prevalent during the time of
the epizootic and cited additional evidence, including high temperature,
debilitation of the fish, and very low water flows as factors in the
reported and other outbreaks of the disease.

Further evidence of the role of bacteria as etiological agents in
epizootics can be cited.  Vibrio sp. (Ross, 1970; Cixar and Fryer,
1969; Smith, 1961; Anderson and Conroy, 1970); Aeromonas salmonicida
(Fryer and Conrad, 1965; Lange and Ljungberg, 1962); Pseudomonas sp.
(Ross, Nordstrom, Bailey and Heaton, 1960; Meyer and Collar, 1965;
Seaman, 1951; Wolf, 1937; Van Duijn, 1938); Pasterurella sp. (Snieszko
et. al., 1964); Streptococcus (Robinson and Meyer, 1966) and Nocardia
sp. (Snieszko, et. al., 1964) have all been shown to be pathogens of
fish.  With the intensified interest in the culture of marine and
freshwater fishes, it is certain that additional organisms will be
added to the list.

Outbreaks of bacterial disease are seldom the result of a single
factor.  Three factors are involved in every potential disease
situation ... susceptible hosts, pathogenic organisms, predisposing
environmental conditions.  All must be present if an outbreak is to
occur.  Snieszko (1964) lists decreased immunological resistance,
poor genetic resistance, temperature stresses, pollution, unfavorable
water chemistry, and unfavorable environmental conditions as some
of the predisposing factors.  In the latter, I would include such
factors as crowding, inadequate food supply, spawning activity,
storms, and seasonal changes.  While bacteria may be the ultimate
cause of death in a particular situation, it is not unlikely that
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some other factor is more important.  Consider, for example, the
massive losses of threadfin shad and Tilapia which occur when the
water temperatures fall below the optimum for these species.  A
sudden cold wave may result in massive losses of these fish.
Survivors or moribund individuals will often yield heavy cultures
of bacterial pathogens and, unless the observer is alert to the
circumstances involved, a diagnosis of a bacterial epidemic might
be given.  Intrusions of salt water into freshwater environments
(or vice versa) can cause similar situations.

Fungi have been indicated in a great many epizootics.  Because of
the obvious nature of fungal growths, many workers are tempted to
list the fungus as the cause of death.  It is my opinion that, for
the most part, such growths are secondary infections feeding upon
necrotic tissue induced by bacterial infections, wounds, or death.
One species, Ichthyophonus hoferi, has reportedly caused major
losses of marine herring (Sinderman, 1958).  Since this organism
is highly contagious, its introduction into new environments would
be expected to lead to epizootics in other natural waters.

Protozoan parasites have been implicated in a number of epizootics
but the occurrence is relatively rare.  One of the most spectacular
outbreaks was that of Ichthyophthirius in the Coosa River of Alabama
in which many species of fish were killed along a 200 mile stretch
of the river (Allison and Kelly, 1963).  Myxosporidia of the genus
Thelohanellus have caused severe mortalities of Coregonus and
Rutilus sp. in Europe (Kudo and Neglitsch, 1971).  Microsporidia
of the genus Glugea were implicated by Putz, Hoffman, and Dunbar
(1965) in an epizootic in gizzard shad in Ohio.  Additional cases
can be cited, but, for the most part, either Ichthyophthirius
(Elser, 1955; Hopkins, 1959; Kudo, 1934) or sporozoans (Sanders,
Fryer and Gould, 1970; Hoshina, 1952; Shulman, 1966) have been
involved.  Other protozoans, including Costia, Chilodonella,
Trichodina, Trichodine11a, etc., cause major problems on fish farms
and hatcheries (Meyer, 1970).  Since these organisms have their
greatest impact on young of the year and small-sized individuals,
it is likely that most epizootics induced by these organisms go
unnoticed.

Metazoan parasites are occasionally involved in epizootics.  As a
rule, however, documented cases are infrequent and this group can
not be considered as a major factor in the initiation of epizootics
even though this group possesses adequate potential for doing so.
Those cases in which they have been involved have usually involved
peculiar conditions which especially favored survival of parasites
far beyond normal circumstances.
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Petrushevski and Shulmari (1958) reported that the introduction of a
monogenean, Nltzchia sturinois, caused major losses of sturgeon in
Russia.  In fish culture, monogenetic trematodes are a serious
problem, posing a threat to the survival of small fish.  Tt is
therefore also possible that the effect of this group of parasites
is over-looked in natural waters.  Large numbers of young of the
year fishes could die without being noticed by most field biologists
and fishermen.

Adult digenetic trematodes have not been identified as causative
agents of mortalities among fish.  However, cercariae, the larval
stages of such flukes, have caused epizottic losses.  Hoffman (1954)
and Krull (1934) both reported losses of fish caused by the penetration
of excessive numbers of cercariae.  Experimental evidence (G. L.
Hoffman, personal communication) suggests that fry and other small
stages of fish may be killed by cercarial penetration but no studies
of such effects in natural waters have been reported.

Cestodes, nematodes, and acanthocephalans are often found in large
numbers in fish.  Often, heavily-infected fish will appear in good
health and show no outward signs of ill effects.  In other instances,
infected fish will be emaciated and debilitated.  Larval cestodes
have caused losses when excessively abundant (Petrushevski and
Shulman, 1958; Moore, 1925) but they are not likely causes of
epizootics in natural waters.

Parasitic copepods have caused major epizootics in a number of areas.
Argulus biramosus killed large numbers of fish in lakes in South
Dakota (Allum and Hugghins, 1959).  Ergasilus arthrosis was the
cause of a heavy infestation of centrarchid fishes stocked in a
brackish water lake in Alabama (Kelly and Allison, 1965).
Ergasilus sieboldi annually kills large numbers of fish in Russia
(Petrushevski and Shulman, 1958).  Although Lernaea cyprinacea
causes major economic losses on fish farms, it is seldom a cause
of death in natural waters (Meyer, 1966).

Leeches have not proved to be a serious problem to date.  The only
reported epizootic in natural waters due to leech infestation
concerns a report by Rupp and Meyer (1954) where brook trout were
killed by an unusually heavy leech population in a lake in Maine.

Occasionally, the mutual occurrence of heavy infections of several
parasites on a particular species of fish may prove fatal.  Petrushevski
and Shulman (1958) report an instance in which they felt an epizootic
in sticklebacks was cuased by a double infection of Gyrodactylus (a
monogenetic trematode) and Glugea (a microsporidian protozoan parasite).

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       PROCEDURES FOR IDENTIFYING THE CAUSE OF A FISH KILL

Field Observations:  Care should be taken to record as much infor-
mation concerning the kill as possible ... seemingly unimportant
items about the hours of the day when it occurred, the weather,
water conditions, and other environmental factors may provide
valuable insight as to the WHY of a kill.  Changes in water color,
algal blooms, vegetation, odor, or flow rates should be noted.
Keep in mind that stress of some kind may have been the predisposing
force.

Observers should study the species make-up of a fish kill with care.
Note the species affected, life stages affected, size of fish
affected, general condition of the affected population, age affected,
number affected, loss pattern, a:id any behavioral patterns exhibited
by moribund fish.

Collection of Samples:  In selecting specimens for study, it is
always desireable to use moribund fish, if possible.  If not, choose
freshly-dead specimens.  Live fish from an affected population are
always desireable to fish which have been dead for some time since
parasites may leave a dead fish and saprophytic bacteria rapidly
infiltrate dead tissue.  If live fish cannot be transported to the
laboratory, the best procedure is to place individual fish in
plastic bags and refrigerate them on wet ice.  If necessary, the
specimens can be frozen but many parasites may be lost during the
freezing and thawing process.  If neither refrigeration or freezing
is possible, the fish may be preserved in 10% formalin or a suitable
fixative as a last resort.  Preservation destroys the usefulness of
the specimens, however, for bacteriological or virological culture
so this procedure should be avoided if possible.

Procedures for Viral Study:  Since few laboratories are equipped
with qualified personnel or equipment for virological studies, it
is adviseable to send suspect specimens to laboratories which have
the necessary facilities.  Frozen specimens are adequate for viral
studies.  The Eastern Fish Disease Laboratory, Kearneysville, West
Virginia; Cooperative Fish Disease Project, Auburn University,
Auburn, Alabama and the Western Fish Disease Laboratory, Seattle,
Washington are some of the stations to which suspect fish may be
sent.  In each case, however, it is adviseable to call prior to
shipping spec Lmens.

Procedures for Bacteriological Study:  When collecting the fish,
examine the external surface of the fish for lesions such as ulcers,

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fin rot, or patches of fungi.  If these are found, a scraping of
tissue from the margin of a lesion should be prepared as a wet mount
(a piece of tissue in a drop of water and covered with a cover slip)
and studies under 440X magnification.  If myxobacteria are present,
they may be observed as long as flexible bacteria are attached to the
tissue by one end.  Frequently, they can be observed moving actively.
See Davis (1953) for a good description.

Material for culture should not be taken until the external surface
of the fish has been sterilized with a good disinfectant such as
household bleach, Roccal, Hyamine, or a similar product.  Using
sterile instruments, the body cavity should be carefully opened
and the internal organs exposed and moved to one side.  Care should
be taken to avoid puncturing or cutting the intestine since leakage
may contaminate the body cavity.  A sterile loop should then be
used to collect tissue from the kidney for inoculating culture medium.
The kidney is the organ of choice but isolates may also be taken
from the liver, blood, and directly from the site of a lesion.
The most commonly used medium for the initial isolation is Tryptic
Soy Agar (TSA), although Beef Heart-Infusion Agar, Blood Agar or
similar rich medium would suffice.  If myxobacteria are suspected,
it is desireabJe to also inoculate some Ordal's medium since
myxobacteria do not grow readily on TSA.  Other specialized media
may be required if the pathogen Involved is not a standard warmwater
fish pathogen.  If bacteria are the likely cause of an epizootic,
workers should expect to find large numbers of a single type of
colony.  If this is the case, it is desireable to transfer the
organism to fresh media (screw cap test tubes filled with slants
of the isoldation medium) and then take or ship the culture to a
bacteriological laboratory, where biochemical and serological tests
required for the identification of bacteria can be run.  Do not
tighten the cap for shipment.

Procedures for Parasitolpgjical Study:  Fish specimens should be
examined for parasites as soon as possible.  External parasites often
leave the body soon after a fish expires, others will die quickly
if the fish is allowed to dry or if the specimen is frozen.  Examine
the external surface of the fish for lesions, cysts, or large organisms
which may or may not be moving.  Study scrapings from lesions, cyst
contents, and large organisms in wet mounts beginning with the
lowest magnification available and work up to the highest magnification
that the thickness of the slide will permit.  For cyst contents, it
may be necessary to use 440X magnification.

In the case of small fish, an entire gill arch may be prepared as
a wet mount.  In large fish, a scissors may be used to remove a
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small number of gill filaments for making the preparation.  Study
all organisms, cysts, and cyst contents as above.

Fin tissue and/or scrapings from the dorse-lateral area of the body
should also be prepared as wet mounts and studied in a similar manner.
Observers should watch especially for movement since most parasites
are active.  If they are the causative agent, large numbers of them
should be present on a high percentage of the affected fish.

After examining the external surface of the fish, open the body
cavity and examine the internal organs.  If the number of specimens
is limited, the following order of procedure should be observed:
(1) collect tissues for the detection of external parasites,
(2) sterilize the external surface of the fish, (3) open the fish
with sterile instruments and collect material for bacterial cultures,
(4) perform the examination of internal organs for possible parasitism.
A failure to observe this sequence may render the material unfit for
a complete examination.

Examine all internal organs for cysts, hemorrhagic areas, blood clots,
lesions, or gross abnormalities.  Carefully remove all suspect tissues
and organs and place them in physiological saline (0.85%) to prevent
them from drying out.  Then examine each under a dissecting microscope,
using the compound microscope if needed to check cyst contents,
organisms, or peculiar structures.   Preserve specimens and tissue
for later study.

If the supply of fish permits, preserve additional specimens to
supply material for further study if needed.

Fixatives and preservatives are available for most of the major groups
of parasites.  Unfortunately, few of them are universally acceptable
or available.  I suggest that field workers carry as many of the
following solutions with them as possible:  10% formalin, Bouin's
solution, AFA (FAA), and 70% ethanol.  Material for the study of
possible protozoan parasites should be dropped in Bouin's or AFA.
Worm parasites should be flattened under a cover slip and then
placed in AFA, parasitic copepods should be placed directly in 70%
ethanol.  Tissues which are suspected of containing parasites of
unknown identity should be placed in Bouin's or AFA.  If only one
fixative or preservative can be carried, 10% formalin may be an
acceptable compromise.  This solution, however, is a very strong
dehydrating agent and may cause excessive constriction or hardening
of specimens.
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For identification purposes, workers need not go beyond genus
identification in most cases.  Hoffman (1967) provides a useful
tool for the general identification of fish parasites.  If precise
identification is desired, smears of the affected organs should be
made.  These smears, small chunks of affected tissue, and specimens
of the parasites should be placed in an appropriate fixative and
preserved for later study.  In many instances, species identification
will be possible only by experienced workers who may use special
stains or equipment in their work.  Most field biologists will
find it adviseable to send the material to some specialist for
identification.  However, I strongly advise against sending every
specimen you encounter since the time of such men is limited and
often too valuable to be requested except when you suspect the
organism to be the cause of a major problem.  Advice on where to
send specimens is available from our laboratory, the Eastern Fish
Disease Laboratory, Western Fish Disease Laboratory, Auburn
University, and zoology departments of most universities.

                           DISCUSSION

Before attempting to ascribe an epizootic to a particular cause,
every observer should study the entire environmental picture to
acquaint himself with as much information about the outbreak as
possible.  Seemingly insignificant factors of weather, water flow,
nutrition, pollution, and water chemistry may play important roles
in the true cause of the epizootic.  As an example, I would like to
describe a unique situation which occurred during the winter of
1963.  Weather conditions showed an abrupt change from a daily
temperature of 25° F to 85° F.  White catfish being held in a 0.1
acre pond at the Fish Farming Experimental Station fed heavily on
chironomid larvae on the third day of warm weather.  During the
following night, the temperature fell from 83° during the day to
about 24° by the following morning.  The change was accompanied
by a brisk wind which speeded cooling of the pond waters.  During
the next day, large numbers of catfish fingerlings were observed
belly-up on the water surface, some in rigor, others making feeble
attempts to swim.  All had distended abdomens and were unable to
dive below the surface.  Losses had begun.  Preliminary symptoms
suggested dropsy (a bacterial disease) but a more complete examination
revealed that the stomach was filled with gas ... the fish were in
fact bloated.  The sudden drop in water temperatures was apparently
so rapid that the fish were unable to digest the contents of the
stomach and gut.  Bacterial decay then began and generated sufficient
gas to bring the fish to the surface.  Transfer of the fish to warmer
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water eliminated some of the problem but both parasitic and bacterial
infections followed the stress of the sudden temperature change and
handling.  Similar relationships are common in natural environments.

Frequently the first indication that something is wrong in natural
waters is the appearance of dead fish.  Such evidence is "after the
fact" and workers are then faced with re-constructing the situation
which led to the kill.  Fish killed by pesticide will appear much
the same as fish killed by an oxygen depletion.  As a consequence,
no evidence should be overlooked.

An epizootic due to parasites can be expected to have an extended
course with fish dying over a prolonged period.  Losses affect a
limited number of species.   Specimens may appear weak or listless
and often will be emaciated.  Small fish would be expected to die
first.  Generally, the die-off will affect a few fish at the outset
and the mortality rate will climb with time.  Other aquatic animals
are not affected.

Bacterially-induced epizootics will follow a similar course of
action.  All sizes of fish will be affected but will usually be
limited to one species.  Occasionally tadpoles and frogs may also
be killed.  As a rule, however, the observer should expect to see
lesions on the body of the fish.  Should an unusually virulent
strain of bacteria be the cause of the epizootic it is possible
that the course of the disease will be very short, highly lethal,
and lack the typical lesions.  Such cases are rare.

Kills resulting from the depletion of oxygen generally occur during
the early morning hours of darkness, but the die-off usually ends
with sunrise.  The mortality will be abrupt and the large specimens
will die first.  Frequently there will be conspicuous changes in the
odor and color of the water.  Vegetation or algal blooms frequently
will also die but zooplankton will be present.  Depending upon the
level to which the dissolved oxygen has dropped, there is often a
species selectivity in an oxygen depletion with those species having
the highest oxygen requirements dying first.

Kills due to toxicants are likewise abrupt.  Mortalities may occur
at any hour and will continue through daylight and darkness.
Zooplankton, snakes, turtles, clams, and frogs may also be affected.
Small fish will die first,  usually exhibiting signs of convulsions,
loss of equilibrium, and toxicosis.  Species selectively will be
evident during the early stages of the kill but, in the case of
highly toxic substances, may progress through an entire population.
Some insecticides cause the fish to exhibit a forward thrusting of

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the pectoral fins (Meyer, 1965), others may cause curvatures of the
spine (Meyer, 1966).  A lethal toxicant such as cyanide may cause
no outward signs but an examination of the internal organs will
reveal massive blood clots.  Water conditions may appear normal
with no apparent color change or odor.
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                        LITERATURE CITED
Allison, R. and H. D. Kelly.   L963.  An epizootic of Ichthyophthirius
     multifiliis in a river fish population.  Progr. Fish-Cult. 25
     (3):149-150.

Allum, M. 0. and E. J.  Hugghins.  1959.  Epizootics of fish lice,
     Argulus biramosus, in two lakes of eastern South Dakota.
     J. Parasitol.  45(4-2):33.

Anderson, J. I. W. and D. A. Conroy.  1970.  "Vibrio disease in
     marine fishes", in Special Publication No. 5.  A Symposium
     on Diseases of Fisheries and Shellfishes.  Amer. Fish. Soc.,
     Washington, D. C.  p. 266-272.

Baier, J. G.  1952.  Ecology of Animal Parasites.  Univ. of
     Illinois Press, Urbana.  224 pp.  (See pp. 6-7).

Bottomley, P. E., and D.  G. Holland.  1966.  Columnaris Disease in
     Coarse Fish.  Association of River Authorities Year Book
     1966, Westminister,  London, S.W.I., England, pp. 101-109.

Chandler, A. C.  1955.   Introduction to Parasitology.  John Wiley
     and Sons, New York.   799 pp.,  (See p. 14).

Cisar, J. 0. and J. L.  Fryer.  1969.  An Epizootic of Vibrosis in
     Chinook Salmon.  Bull. Wildlife Disease Assoc., Vol. 5,
     April, 1969, p. 73-76.

Colgrove, D. J. and J.  W. Wood.  1966.  Occurrence and Control of
     Chondrococcus Columnaris as Related to Fraser River Sockeye
     Salmon.  International Pacific Salmon Fisheries Comm.,
     Progress Report No.  15, New Westminister, B.C. Canada, 1966,
     51 pp.

Davis, H. S.  1953.  Culture and Diseases of Game Fishes.  University
     of California Press, Berkeley and Los Angeles, pp. 265-272.

Elser, H. J.  1955.  An epizootic of ichthyophthiriasis among fishes
     in a large reservoir.  Progr. Fish-Cult.  17:132-133.

Elson, K.G.R.  1968.  Salmon Disease in Scotland.  The Salmon Net,
     The magazine of the Salmon Net Fishing Association of Scotland,
     No. 4, pp. 9-17, June, J968.

Fryer, J. L. and J. F.  Conrad.  1965.  Some observations on
     Furunculosis in adult Pacific Salmon and Steelhead trout.
     Progr. Fish-Cult., Vol. 27(2):99-100.
                                 57

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Haley, R., S. P. Davis, and J. M. Hyde.  1967.  Environmental
     stress and Aeromonas liquefaciens in American  and  threadfin
     shadmortaliti.es.  Progr. Fish-Cult. 29(4) :193.

Hoffman, G. L.  1954.  The cercaria and development  of  Crassiphialia
     (Neascus) bulboglossa (Trematoda.-Strigeida) .   Proc. No. Dak.
     Acad. Sci., Vol. 9:57-58.

	.  1967.  Parasites of North American  Freshwater  Fishes.
     University of California Press, Berkeley and Los Angeles,  486  pp.

Hopkins, C. A.  1959.  Seasonal variations in the incidence and
     development of  the cestode Proteocephalus  filicollis  (Rud. 1810)
     in Gasterosteus aculeatus (L. 1766).  Parasitology 49(3-4):529-542,

Hoshina, T.  1952.  Notes on some myxosporidian parasites  on fish of
     Japan.  J. Tokyo Univ. Fish.  39:69-97.

Kelly, H. D. and R. Allison.   1965.  Observations on the infestation
     of a fresh water fish population by a marine copepod  (Ergasilus
     lizae Kroyer 1863).  Proc. of the 16th Ann. Conf.  S.E. Assn.
     Game and Fish Comm., October 14-17, 1962,  Charleston, S.  Carol.
     pp. 236-239.

Kudo, R. and P. Meglitsch.  1971.  A synopsis of genera and species
     of Myxosporidia.  Univ.  of So. 111. Press, (In press).

Krull, R. R.  1934.  Cercaria bessiae Cort and  Brooks,  1928, an
     injurious parasite of fish.  Copeia 2:69-73.

Kudo, R. R.  1934.  Studies on some protozoan parasites of fishes
     of Illinois.  Illinois Biol. Monogr.  13(1):44 pp.

Lange, J., and 0. Ljungberg.   1962.  Outbreaks  of fish  furunculosis
     in salmonids in Sweden 1951-1960.  Nordisk Veterinaermed.
     14(3):177-191.

Lennon, R. E. and Parker, P.S.  1960.  An outbreak  of columnaris
     disease in stonerollers.  Progr. Fish-Cult. 22(3):102.

Meyer, F. P.  1964.  Field treatments of Aeromonas  liquefaciens
     infections in golden shiners.  Progr. Fish-Cult. 26:33-35.

       	.  1965.  The experimental use of Guthion as  a selective
     fish eradicator.  Trans. Amer. Fish. Soc., 94(3):203-209.

     	.   1966.  A new control for the anchor parasite,
     Lernaea cyprinacea.  Progr. Fish-Cult.  28(l):33-39.

     	.   1970.  Seasonal fluctuations in the incidence  of
     disease on fish farms.  In Special Publication No. 5:   A.
     Symposium on Disease of Fishes and Shellfishes.  Amer.  Fish.
     Soc., Washington, D. C.  p. 21-29.

                                58

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Meyer, F. P. and J. D. Collar.  1964.  Description and  treatment  of
     a Pseudomonas infection in white catfish.  Appl. Microbiol.
     12:201^203.

Moore, E.  1925.  Diseases of Fish.  14th Annual Rept.  of  the
     N.Y. State Conserv. Dept,, p. 83-87.

Pacha, R. E. and J. E. Ordal.  1970.  Myxobacterial diseases of
     Salmonids.  In:  A Symposium on Diseases of Fishes  and Shellfishes,
     S. F. Snieszko, editor.  Amer. Fish. Soc., Washington, D. C.
     243-257.

Petrushevski, G. K. and S. S. Shulmari.   1958.  The parasitic diseases
     of fishes  in the natural waters of  the USSR.  In:   Parasitology
     of Fishes  edited by V. A. Dogiel, G. K. Petrushevski, and
     Yu. I. Polyanski, Oliver and Boyd,  London (1961) pp.  299-319.

Pippy, John H.  C. and Gerard M. Hare.  1969.  Relationship of river
     pollution  to bacterial infection in salmon (Salmo  salar) and
     suckers (Catostomus ^omi^rjsjimT^) .  Trans, of the Amer. Fish.
     Soc., 98(4)^685-690.

Putz, R. E., G. L. Hoffman, and C. E. Dunbar.  1965.  Two  new species
     of Plistophora (Microsporidia) from North American  fish with
     a synopsis of freshwater and euryhaline fishes.  J. Protozool.
     12(2):228-236.

Pyefinch, K. A. and K. C. R. Elson.  1967.  Salmon diseases in Irish
     rivers.  Scottish Fisheries Bulletin No. 26.  pp.  1-4,

Reed, G. B. and G. C.  Toner.  1942.  Proteus hydrophilus infections
     of pike, trout, and frogs.  Canad.  Jour. Res. 20:161-166.

Reichenbach-Klinke, H. and E. Elkan.  1965.  The Principal Diseases
     of Lower Vertebrates.  Academic Press, New York, pp.  55-60.

Ricker, W. E.   1945.  Causes of death among Indiana fishes.  Trans.
     10th No. Amer. Wildlife Conf., Amer. Wildlife Inst., Washington,
     D. C., pp. 267-269.

Robinson, J. A. and F. P. Mey^r.  1966.  Streptococcal  fish pathogen.
     J. Bact. 92(2):511\

Rock, Leo F. and Harold M. Nelson.  1965.  Channel catfish and
     gizzard shad mortalitv caused by Aeromonas liquefaciens.
     Progr. Fi sh-Cu If. 27(3): J 38-1.41.

Ross, Avron J.  1970.   Vibriosis in Fish.  U. S. Dept.  Interior,
     Bur. of Sport Fisheries & Wildlife, FDL-29.  3 p.
                                 59

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Ross, A. J., P. R. Nordstrom, J. E. Bailey,  and  J.  R.  Heaton.   1960.
     A bactetial disease of yellow perch  (Pe_rc_a  flavescens) .
     Trans. Amer. Fish. Soc.  89(3) : 310-312 .


Rupp, R. S. and M. C. Meyer.  1954.  Mortality among brook trout
     (Salvelinus foritinalis) , resulting from attacks of  freshwater
     leeches.  Copeia, No. 4, p. 294-295.


Sanders, J. E., J. L. Fryer, and R. W. Gould.  1970.   Occurrence  of
     the Myxosporidian parasite, Ceratomyxa  shasta, in salmonid
     fish from the Columbia River Basin and  Oregon  coastal streams.
     In Special Publication No. 5:  A  Symposium  on  Disease of  Fishes
     and Shellfishes, Amer. Fish. Soc., Washington, D. C. ,  p.  133-141.


Shulman, S. S.  1966.  The myxosporidean  fauna of SSSR.   Izdatel'stvo
     Nauka, Moscow, 507 pp.


Sinderman,  C. J.  1968.  An epizootic  in  Gulf of Saint Lawrence fishes.
     Trans, 23rd No. Amer. Wildlife Conf., pp. 349-360.


Smith, Isabel W.  1961.  A disease of  finnock due to Vibrio
     anguillarum.  Jour. Gen. Microbiol.  24(2):247-252;  BiolAbstrs.
     36(10).


Snieszko, S. F.  1964.  Remarks on some facets of epizootiology
     of bacterial fish diseases.  Dev. Indust. Microbiol.  5:97-100.


	.  1958.  Columnaris disease of fishes.  Fishery Leaflet
     461, U. S. Dept. of Interior, Bureau of  Sport  Fisheries  &
     Wildlife, 3 p.


Snieszko, S. F., G. L. Bullock, Edgar Hollis,  and J.  G.  Boone.   1964.
     Pasteurella sp. from an epizootic of white perch (Roccus
     americanus) in Chesapeake Bay tidewater  areas.   J.  Bact.,  Vol.
     88(6):1814-1815.


Snieszko, S. F., G. L. Bullock, C. E. Dunbar  and L. L. Pettijohn,   1964.
     Nocardial Infection in hatchery-reared  fingerling rainbow
     trout (S_almo_ ^alr_dne_rj_) .  J. Bact., Vol.  88(6) : 1809-1810.


Wagner, E. I), and C. L. Perkins.  1952.  Pseudomonas  hydrophila,
     the cause of "red mouth." disease in rainbow trout.   Progr.
     Fish-Cult.  14:I?7-128.


Wedemever, G.  1970.  The rc-le of stress in  the disease  resistance
     o L f i s 11 e s .  In:  A_ Svjnpp s i _un_ _on_ I) i s ease, s  of Fishes  and Shellfishes ,
     American Fisheries Society, S.  F. Snieszko, editor,  Washington, D.  C. ,
     D, 30-35.
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             MORTALITIES FROM MISCELLANEOUS CAUSES AND
              IMPORTANT ENVIRONMENTAL CONSIDERATIONS


                                by


                         Ralph M. Sinclair


            Aquatic Biologist, National Training Center
          Office of Water Programs, EPA, Cincinnati, Ohio



 I.  Fish Kills vs. Natural Mortalities


     A.  Natural Mortalities - Those which are caused through natural
         phenomena such as: acute temperature change, storms, ice
         and snow cover, decomposition of natural materials, salinity
         change, spawning mortalities, parasites, and bacterial or
         viral epidemics.


     B.  Man caused fish kills - Produced by environmental changes
         through man's activity, and may be attributed to municipal
         wastes, industrial wastes, agricultural activities and
         water control activities.


         1.   Direct


         2.   Indirect


II.  Organism Interactions


     A.  Pollution is often studied as a factor affecting the biota,
         but it is equally important to recognize the environmental
         changes produced by the biota.  According to Westlake,
         under many conditions ".... the environment is almost as
         much a product of the community as the community is of
         the environment."


     B.  Changes in ecosystems are occurring continuously.


         1.   Myriad interactions take place at every moment of the
             day as plants and animals respond to variations in
             their surroundings and to each other.  Evolution has
             produced for each species, including man, a genetic
             composition that limits how far that species can go
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    in adjusting to sudden changes in its surroundings.
    But within these limits the several thousand species
    in an ecosystem, or for that matter, the millions in
    the biosphere,  continuously adjust to outside stimuli.
    Since interactions are so numerous, they form long
    chains of reactions.


2.  Small changes in one  part of an ecosystem are likely

    to be felt and  compensated for eventually throughout
    the system.  Dramatic examples of change can be seen
    where man has altered the course of nature.   It is
    vividly evident in his well-intentioned but poorly
    thought out tampering with river, lake, and other
    ecosystems.


    a.  The Aswan High Dam


    b.  The St. Lawrence  Seaway


    c.  Lake Karl ha


    d.  The Great Lakes


    e.  ValJev of Mexico


    f.  California  eatthauake (Scientific American 3981,

        P- 333)


    g.  Everglades  and the Miami, Florida Jetport


    h.  CopperhiJl, Tennessee (Copper Basin)


3.  Ecosystem Stability


    a.  The stability of  a particular ecosystem depends on
        its diversity.  The more interdependences in an
        ecosystem,  the greater the chances that it will be
        able to compensate for changes imposed upon it.


    b.  The least stable  systems are the single crops -

        called monocultures - created by man.  A cornfield
        or lawn has Jittle natural stability.  If they are
        not constantly and carefully cultivated, they will
        not remain  cornfields or lawns but will soon be

        overgrown with a  wide variety of hardier plants
        constituting a mere stable ecosystem.
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              c.  The chemical elements that make up living systems
                  also depend on complex,  diverse sources to prevent

                  cyclic shortages or oversupply.


              d.  The oxygen cycle, which  is crucial to survival,

                  depends upon a vast variety of green plants,
                  notably plankton in the  ocean.


              e.  Similar diversity is essential for the continued

                  functioning of the cycle by which atmospheric

                  nitrogen is made available to allow life to exist.

                  This cycle depends on a  wide variety of organisms,
                  including soil bacteria and fungi, which are often

                  destroyed by pesticides  in the soil.


          4.  Some environmental examples:


              a.  Diatom Blooms


                  "••• Asterionella (an oil storage alga) produces an
                  autotoxin (autoantibiosis) which will inhibit itself,

                  may stimulate or inhibit other species that store
                  oil, but always stimulate algae that store starch.

                  For example, Asterionella may produce a bloom and

                  inhibit itself, but stimulate a population of
                  Synedra.  These oil storage algae produce a substance
                  that stimulates a starch-storing alga, Coelastrum,

                  which may stimulate another starch-storing organism,
                  Cosrnarium, and they, in  turn, stimulate an oil
                  storing species of Dinobryon."  Patrick.


              b.  The altered structure of the plankton community
                  due to the introduction  of the alewife.


              c.  Fecal deposit feeders in the estuarine environment.


III.  Biological Evaluation


      A.  Biological evaluation of an aquatic environment includes a
          comparison of the living community at one location with

          that of another.  It silso calls  for an understanding of

          the physical conditions impinging upon that population.


      B.  The inability ot? man to adequately predict or control his

          effects on the environment is indicated by his lack of

          knowledge concerning the net effect of atmospheric pollution

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         on the earth's climate.  Similarly, there are large scale
         fish mortalities, which are associated with red tide
         organisms (dinoflagellates), that may, or may not, be
         related to the discharge of nutrients.


     C.  Levels of Biological Integration


         1.  From higher to lower we have:


             a.  Biosphere


             b.  Ecosystems


             c.  Communities


             d.  Populations


             e.  Individuals


             f.  Organ Systems


             g.  Organs


             h.  Tissues


             i.  Cells


             j.  Protoplasm


         2.  When we observe phenomena at some level, the causes
             (or explanation of mechanisms) are usually to be
             found at a lower level of biological integration and
             the effects (or significance) at a higher level.


IV.  Possible Natural Causes of Fish Kills


     A.  Oxygen depletion due to ice and snow cover on surface waters.


     B.  Oxygen depletion at night because of plant respiration or
         at anytime during the day because of natural occurring
         organics in the water.


     C.  Abrupt temperature changes.


     D.  Epidemic and endemic diseases, parasites, and other
         natural occurring biological causes.
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      E.  Lake water inversion during vernal or autumnal turnover
          which results in toxic material or oxygen-free water being
          brought to the surface.


      F.  Interval seiche movement in which a toxic or low DO
          hypolimnion flows up into a bay or bayou for a limited
          period of time, and later returns to normal level.


  V.  "Natural Populations"


      A.  Man's activities have left few, if any, control (reference)
          populations.   As an example, pesticides and combustion
          products are  now all globally dispersed.


      B.  Most populations are continuously or intermittently stressed
          by man's activities.  If these stresses produce populations
          susceptible to various parasites or diseases, should we call
          this a natural mortality?


 VI.  Pollution Comes in Many Packages


      A.  The sources of air, water, and land pollution are interrelated
          and often interchangeable.


          1.  A single  source may pollute the air with smoke and
              chemicals, the land with solid wastes, and a river or
              lake with chemical and other wastes.


          2.  Control of air pollution may produce more solid wastes,
              which then pollute the land or water.


          3.  Control of wastewater effluent may convert it into solid
              wastes, which must be disposed of on land.


          4.  Some pollutants - chemicals, radiation, pesticies -
              appear in all media.


      B.  "Disposal" is as important and as costly as purification.


VII.  Persistent Chemicals in the Environment


      Increasingly complex manufacturing processes,  coupled with
      rising industrialization, create greater amounts of exotic
      wastes potentially toxic to humans and aquatic life.
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They may also be teratogenic (toxicants responsible for
changes in the embryo with resulting birth defects, ex.,
thalidomide), mutagenic (insults which produce mutations,
ex., radiation), or carcinogenic (insults which induce
cancer, ex., benzopyrenes) in effect.

A.  Metals - current levels of cadmium, lead, and other
    substances whose effects on humans and fish and wildlife
    are not fully understood constitute a mounting concern.
    Mercury pollution, for example, has become a serious
    national problem.

B.  Pesticides

    1.  A pesticide may move through an ecosystem in many ways.
        Hard (pesticides which are persistent, having a long
        half-life in the environment includes the organochlorines,
        ex., DDT) pesticides ingested or otherwise borne by the
        target species will stay in the environment, possibly
        to be recycled or concentrated further through the
        natural action of food chains if the species is eaten.
        Most of the volume of pesticides do not reach their
        target at all.

    2.  Biological magnification

        Initially, low levels of persistent pesticides in air,
        soil, and water may be concentrated at every step up
        the food chain.  Minute aquatic organisms and scavengers,
        which screen water and bottom mud having pesticide
        levels of a few parts per billion, can accumulate
        levels measured in parts per million - a thousandfold
        increase.  The sediments including fecal deposits are
        continuously recycled by the bottom animals.

        a.  Oysters, for instance, will concentrate DDT 70,000
            times higher in their tissues than its concentration
            in surrounding water.  They can also partially
            cleanse themselves in water free of DDT.

        b.  Fish feeding on lower organisms build up concentrations
            in their visceral fat which may reach several thousand
            parts per million and levels in their edible flesh
            of hundreds of parts per million.
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        c.  Larger animals, such as fish-eating gulls and other
            birds, can further concentrate the chemicals.  A
            survey on organochlorine residues in aquatic birds
            in the Canadian prairie provinces showed that
            California and ring-billed gulls were among the
            most contaminated.  Since gulls breed in colonies,
            breeding population changes can be detected and
            related to levels of chemical contamination.
            Ecological research on colonial birds to monitor
            the effects of chemical pollution on the environment
            is useful.

C.  "Polychlorinated biphenyls" (PCB's)

    PCB's are used in plasticizers, asphalt, ink, paper, and
    a host of other products.  Action has been taken to curtail
    their release to the environment, since their effects are
    similar to hard pesticides.

D.  Multifactoral Mortalities

    When some stress is placed on a population they become
    particularly susceptible to a pathogen and/or parasite.
    Thus, we have one or more primary stress events which
    lead to secondary causes of death.

    1.  A serious disease of salmonids in the United Kingdom
        is Ulcerative Dermal Necrosis (UDN).  A strain of
        Saprolegnia has been indicated as cause for death,
        but investigators are divided as to the real causitive
        factor(s).

    2.  Factors involved would include:

        a.  Species

        b.  Age

        c.  Breeding condition

        d.  Pre-disposition to the disease

        e.  Chemical environment

        f.  Physical environment

        g.  ImmunoJogical barriers

        h.  Exposure to pathogens

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VIII.  Other Sources of Stress in Populations - Biological Pollution

       Contamination of living native biotas by introduction of exotic
       life forms has been called biological pollution by Lachner et al.
       Some of these introductions are compared to contamination as
       severe as a dangerous chemical release.   They also threaten to
       replace known wildlife resources with species of little or
       unknown value.

       A.   Tropical areas have especially been vulnerable.  Florida
           is referred to as "a biological cesspool of introducted
           life".

       B.   Invertebrates

           a.  Asian Clams have a pelagic velier larvae, thus, a
               variety of hydro installations are vulnerable to
               subsequent pipe clogging by the adult clams.

           b.  Melanian snails are intermediate hosts for various
               trematodes parasitic on man.

       C.   Vertebrates

           a.  At least 25 exotic species of fish have been established
               in North America.

           b.  Birds, including starlings and cattle egrets.
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                           REFERENCES
Environmental Quality.  Second Annual Report of the Council on
    Environmental Quality.  August 1971.


Lachner, Ernest A., Robins, C. Richard, and Courtenay, Walter R. Jr.
    Exotic Fishes and Other Aquatic Organisms Introduced into
    North America.  Smithsonian Contrib. to Zool.  59:1-29.  1970.


Miller, Morton W. and Berg, George C.  Chemical Fallout; Current
    Research on Persistent Pesticides.  531 p.  Thomas.  1969.


Sondheimer, Ernest B. and Simeone, John B.  Chemical Ecology.
    Academic Press.  336 pp.  1970.


Whittaker, Robert H.  Communities and Ecosystems.  Macmillan Co.
    162 pp.  1970.


U. S. Geological Survey.  Mercury in the Environment.  Geol. Surv.
    Prof. Paper T13.  67 pp.  1970.


Toxic Substances.  Council on Environmental Quality.  25 p.
    April 1971.


Zinc in Water.  A Bibliography USDI.   Office Water Resources WRSIC
    Series 208.  1971.  Also in this series WRSIC 201-207; Mercury,
    Magnesium, Manganese, Copper, Trace Elements, and Strontium.


Willoughby, L. G.  Salmon Disease in Windermere and the River
    Leven; The Fungal Aspect.  Salmon and Trout Magazine.  186:124-130.
    1969.
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                   TOXICANT CAUSED  FISH KILLS
   COLLECTION, PRESERVATION, AND ANALYSES  OF INORGANIC  SAMPLES
                                hv
                          Mark  Colernan
               Supervisor, Water Quality  Laboratory
        State Department of Health, Oklahoma  City, Oklahoma
                      I INTRODUCTORY  REMARKS

As a representative of the primary  analytical  laboratory  for water
analyses in Oklahoma, I should  like  to point out  that  we  are aware
that, in the particular case of  fish kills, we  are  only looking  at
the top part of the iceberg.  We realize  that  although fish  may
not be actively and openlv dying in  a particular  body  of  water,
the quality of that water may be changing in such a way as  to cause
a deleterious effect on the fish populations.

When we were asked to participate in this seminar,  we  were  told  that
we would primarily be speaking  to administrators  as opposed  to
actual investigators.  This will prejudice our  remarks somewhat.

Up to this point, you have heard about sophisticated teams  and
sophisticated methods to investigate fish kills.  You  have heard
references to teams composed of bio] >gists, chemists,  and engineers.
It might be adviseable to note  that  even  the EPA  does  not have such
a team in this region,  i\e > t he r  do we have such a team in Oklahoma.
This talk is, therefore, on a somewhat Lower plane  than have been
some of the previous talks.

                        SAMPLE  COLLi-ClTON

On numerous occasions throughout tills seminar,  time has been
emphasized as one of tht.- must important factors in  the investigation
of a fish kiJl.  It :.- essential that representative samples be
taken from active problem ureas.  Without samples from active
problem areas, it is extreme!1/  difficult, and  frequently  impossible,
to determine the canst •:•'  i fish kill.  Samples should also  be
taken from all potential sources v'irhin the area  that  could  contribute
to the problem.  Each tributary  should be sampled.  It is also
extremely desirable to h,ive bnnples  which are  above and below the
problem area as well a.- ab, -A  ,uic! !•< low the probable source  or
sources of the problerr,  ['he purpose of the above and  below  samples
is to provide comparative uat i  for  the determination of why  a
particular water i -; ki 1 ' '':>.;. ; is<,.

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A natural question  that arises  is  relative  to  the number  of  samples
which should be taken.  I do not fee]  as  if  a  set number  of  samples
should be taken.  It is extremely  important  that your  investigator
be a capable man who will utilize  good judgement and take those
samples which are absolutely necessary to define the problem and
eliminate potential sources so  that the actual primary  offender
may be determined.  We have found  that it is adviseable for  the
analyst to be included on this  survev  team.  (Of course,  in  our
case, frequently the entire team is the analyst).   It  is  extremely
important that your investigators  only take  those samples which are
necessary.  It is not infrequent that  inexperienced investigators
will take unnecessary samples merely to prove  that  they were at
a particular point.  This doe?-, of course, help  to  justify travel
expenses.  However, we are not  in  the business of justifying travel
expense, rather, we are attempting to  investigate a fish  kill.

I would like to reemphasize the point  that samples  should be taken
where and while the fish are actually  dying.   Samples which  are
taken after the fact, are generally taken after  the problem  has
washed downstream.  An early morning dissolved oxygen  test is
frequently highly desirable.  In any case where  a depletion  of
oxygen is suspect as the cause  of  the kill,  or in cases where a
marginal dissolved oxygen content  is noted,  useful  information may
be derived from a dissolved oxygen determination immediately prior
to sunrise.

                       SAMPLE PRESERVATION

When one considers  the amount of money which is  invested  in  each
sample, it becomes apparent that ail steps must be  taken  to  insure
that those samples be as representative as possible of  the conditions
existing at the time of sampling.  Hopefullv,  containers  for the
sample will be supplied hv the  1 ;ib<-r,'it -TV .   Unfortunately, in
Oklahoma, we do not have sufficient fand^ to provide such containers
to all personnel who might possibly take samples associated  with
fish kills.  Of course, sample bottles are supplied for our  own
personnel.  If a sample bottle supplied hv  the laboratory is not
available, one should Litili^e at, clean 3 bottle  as  possible  to take
the sample.  In the past, wi- have  suggested  that a  bottle of
distilled water be purrha- ed f'T use as a sample container.  Whatever
container is used,  the ionr,unrr should be  flushed  with the  water
to be sampled several tiroes.  Our  preferance is  for plastic  containers
with deep-sealing air-tight non-metallic caps.  We  ask  for at least
a half gallon sample, dm- to the fact  that  frequently we  do  not
know the chemical cv:.usi  .if U,..   fist- kill  and must make numerous
analytical de terminal i 01-5 ,

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If the sampler has a fairly accurate idea of which  chemical
parameter is causing the fish kill, it is highly desirable to
preserve the sample according to the enclosed  chart taken from
the 1971 Environmental Protection Agency Manual for Chemical
Analyses of Water and Wastewater.  Unfortunately, such preservatives
frequently are not available.  Should this be  the case, one should
refrigerate all samples and deliver those samples immediately to
the analyst.  In cases where samples are preserved,  additional
samples which are not preserved should be taken and refrigerated.

It is imperative that in all cases the samples be delivered as
quickly as possible to the analyst.

               SAMPLE ANALYSIS FOR INORGANIC CAUSES

A great deal of the analytical work, surrounding a fish kill is
determined by the field investigation.  It is  extremely important
that dissolved oxygen he determined at the site at  the same time
as the samples are tak^n.  It is also highly desirable that
temperature, pH, total alk.^.lini tv, and other parameters (such as
ammonia) be determined dependent on the type of kill.  If the
field investigator is able to get a rough idea of the number of
fish and the numbers of each species which are affected, the types
of industries, and whether or not a municipality discharges in
the area, much analytical time can be saved.   (As a general rule
of thumb, if the fish Kill tends t.< involve a  single species,
we tend to believe that a kill o1 a biological nature is in
progress.)  If a municipality is on the water  shed  and a high
dissolved oxvgen is noted, we tend to rule out sewage as a cause
of the kill, and su ou.  In i>!her words, the field  investigation
rules out or rules in various analytical approaches.

All methodology should follow Standaid Methods as outlined in
the EPA manual for analvtic.nl techniques.  Dependent on the type
of kill, parameters such as COD, BOD^, ammonia, pll,  total alkalinity,
chloride, metals, and ,>l~her analyses are determined, again,
dependent on the field observations.  If the field  investigation
does not tend to single out a particular type  of waste as being
the cause, as a last resort, we- run everything and  hope to find
some causative agent.

                       CONCLUDING REMARKS

It might be wise to point out that we are not  in the business of
soliciting samples.  We have sufficient samples at  any given time.
Additionally, samples whirl: are or are not taken properly, or

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which are or are not necessary, but which relate to a fish kill,
have a very high priority.  As a result, it is necessary to run
these samples in lieu of other samples.  Generally, the other
samples would be drinking water supplies.  It is important to
point out that samples should not be taken unless an extremely
good reason exists for analytical results of that sample.
                                73

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                           REFERENCES


Jenkins, David.  "A Study of Methods Suitable for the Analysis and
     Preservation of Phosphorus Forms in an Estuarine Environment".
     Report for the Central Pacific River Basins Project, Southwest
     Region, FWPCA (1965).


Jenkins, David.  "A Study of Method for the Analysis and Preservation
     of Nitrogen Forms in an Estuarine Environment".  Report for the
     Central Pacific River Basins Project, Southwest Region, FWPCA
     (1965).


Howe, L. H. and Holley, C. W.  "Comparisons of Mercury (II) Chloride
     and Sulfuric Acid as Preservatives for Nitrogen Forms in Water
     Samples".  Env. Sci. & Tech. 3:478 (1969).


*"Methods for Chemical Analysis of Water and Wastes, 1971".  Environmental
     Protection Agency, Analytical Quality Control Laboratory,
     Cincinnati, Ohio.
                                 74

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                          Table 2 - Sample Preservation*
    Parameter


Acidity-Alkalinity


Biochemical Oxygen Demand


Calcium


Chemical Oxygen Demand


Chloride


Color


Cyanide


Dissolved Oxygen


Fluoride


Hardness


Metals, Total


Metals, Dissolved


Nitrogen, Ammonia


Nitrogen, Kjeldahl


Nitrogen, Nitrate-Nitrite


Oil and Grease


Organic Carbon


PH


Phenolics




Phosphorus
    Preservative


Refrigeration at 4°C


Refrigeration at 4°C


None required


2 ml H2S04 per liter


None required


Refrigeration at 4°C


NaOH to pH 10


Determine on site


None required


None required


5 ml HNO  per liter
                                    Maximum
                                 Holding Period


                                   24 hours


                                    6 hours


                                    7 days


                                    7 days


                                    7 days


                                   24 hours


                                   24 hours


                                   No holding


                                    7 days


                                    7 days


                                    6 months
Filtrate: 3 ml 1:1 HNO  per liter   6 months
40 mg HgCl * per liter - 4°C


40 mg HgCl * per liter - 4°C


40 mg HgCl * per liter - 4°C


2 ml H2S04 per liter - 4°C


2 ml H2S04 per liter (pH 2)


Determine on site


                  ?04 to


  pH 4.0 - 4°C


40 mg HgCl * per liter - 4°C
1.0 g CuS04/l +
 7 days


Unstable


 7 days


24 hours


 7 days


No holding


24 hours




 7 days
'Disposal of mercury-containing samples is  a recognized problem;  research
 investigations are under way to replace it as  a preservative.
                                        75

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1
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1

1



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•

1



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1
1
Table 2 - Sample Preservation


Parameter
Solids
Specific Conductance

Sulfate
Sulfide
Threshold Odor
Turbidity


References :
Jenkins, David, "A Study
servation of Phosphorus
for the Central Pacific
FWPCA (1965) .
Jenkins, David, "A Study
of Nitrogen Forms in an
(Continued)

Preservative
None available
None required

Refrigeration at 4°C
2 ml Zn acetate per liter
Refrigeration at 4°C
None available




Maximum
Holding Period
7 days
7 days

7 days
7 days
24 hours
7 days
.


of Methods Suitable for the Analysis and Pre-
Forms in an Estuarine Environment." Report
River Basins Project, Southwest
Region,
of Method for the Analysis and Preservation
Estuarine Environment." Report
Central Pacific River Basins Project, Southwest Region,
Howe, L. H. and Hoi ley, C
for the
FWPCA (1965) .
. W. "Comparisons of Mercury (II) Chloride
and Sulfuric Acid as Preservatives for Nitrogen Forms in
Samples." Env. Sci. §

^"Methods for Chemical Ana
Environmental ProteelL
Cincinnati, Oliio



Tech. 3:478 (1969).

lysis of Water and Wastes, 1971"
Water



cm Agency, Analytical Quality Control Laboratory



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                    TOXICANT CAUSED FISH KILLS

     COLLECTION, PRESERVATION, AND ANALYSIS OF ORGANIC SAMPLES


                                by


                           Larry Streck


            Chemist, Surveillance and Analysis Division

                  Environmental Protection Agency

                           Ada, Oklahoma




                            COLLECTION


Fish kills can result from a variety of causes such as:


     1.  Low dissolved oxygen concentrations


     2.  Toxic chemicals


     3.  Excessive water temperatures or rapid changes in water

         temperatures


     4.  Extreme pH levels


     5.  Excessive suspended solids


     6.  The effects of diseases and parasites, etc.


When investigating a fish kill, all of the possible causes should
be kept in mind.  Any factors that might cause a change in water
conditions should also be considered.  For example, a heavy rainfall
shortly after a pesticide application may wash sufficient pesticides
into a stream to cause a fish kill, or an extended drouth with high
temperatures might reduce the flow of dilution water producing toxic
results from a normally acceptable industrial effluent.


Sampling sites and the number of samples to take depends on several
factors.  In a flowing stream it is necessary to take water samples
in the area of the fish kill, above it, and below it.  Since the

polluting substance may have been present in a section of the stream,

it may be advisable to collect samples at a distance downstream from

the central kill area which corresponds to the elapsed flow time
since the kill was first noticed.  When fish kills occur in lakes

or impoundments, the depth of sampling is important because of
stratification.
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An important aspect in sample collection is that of the on-site
investigation.  Information concerning an industry's products as
well as by-products could be valuable to help narrow the possibilities
and cut the work load of the analyst.  If pesticides are suspected,
a list of possibilities would be appreciated.  The usual procedure
is to approach the chemist, with samples in hand, and ask for - yea
demand - a complete pesticide analysis.  It is important to realize
that, even though proper samples were taken and preserved correctly,
if the on-site investigation was not conducted properly, generally
no amount of laboratory investigation can solve the fish kill.  The
amount of useful laboratory work that can be done is often limited
by what was not done during the on-site investigation.

Before the actual collection, all containers to be used for organic
samples, should be prepared by rinsing with running tap water,
distilled water and several times with a suitable redistilled
solvent (e.g., acetone, hexane, petroleum ether, chloroform).  Caps
and liners are washed with detergent, rinsed with tap water, distilled
water and solvent.  Some also recommend heat treatment at 300°C
overnight to destroy any trace of organic matter.

Assuming competent sampling personnel, from one liter to one gallon
of water is collected in glass bottles equipped with screw caps
fitted with Teflon liners.  Plastics, such as polyethylene, are not
suggested because traces of plasticizer can be leached from the
plastic by the water and can be a source of analytical interference.
If Teflon liners are not available, clean solvent rinsed aluminum
foil may be used.  Some plastics may actually absorb pesticides
and other organic materials, so its use is certainly discouraged.
Many investigators avoid the use of glass sample bottles because
breakage in shipment frequently causes loss of sample.  This can
be overcome by the use of expanded polystyrene foam shipping containers
molded to fit the bottle.

                           PRESERVATION

Whatever the method of preservation used for organic samples, it
is generally intended to:

     1.  retard biological action

     2.  retard hydrolysis of chemical compounds and
         complexes and

     3.  reduce volatility of the constituents.
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Although a universal method or additive for the preservation of
samples would be nice, it is a practical impossibility.  At best,
the various procedures for preservation can only slow the inevitable
chemical and biological changes which occur when a sample is removed
from a stream or lake.

Techniques for preserving organic contaminated samples are usually
limited to solvent addition, refrigeration, and freezing.  Needless
to say, the length of storage should be as short as possible.  Even
properly preserved samples often have short holding times.  EPA's
Methods for Analysis recommends the following holding times:

              Oil and Grease      24 hours

              Phcnolics           24 hours

              Organic Carbon       7 days

The wide variety and large number of pesticides in use today makes
it impossible to specify a time beyond which all analyses are
worthless or at best questionable.  However, it can be stated that
a practical time limit should be measured in days and in some
instance to hours.  Some of the phosphates, carbamates, and 2-4-D
esters will even undergo slow hydrolysis at a neutral pH and low
temperature.  Because pesticide samples are subject to change, it
is important that dates of analyses be included in any report so
proper interpretations can be made.  Although not applicable to
all types of samples, immediate refrigeration at temperatures near
freezing or below is the best preservation technique available,
at least until the advice of laboratory personnel can be obtained.

                             ANALYSES

Realizing that most laboratories are searching for the minimum
amount of time and effort required to complete a project, one
should consider two questions:

     1.  Must the laboratory investigation start from scratch, or
         did preliminary work show that there was one or more major
         suspects responsible for the fish kill?

     2.  How well must the probable cause be established?
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It is quite difficult to determine the number of samples that can be
handled, the cost involved, and the time required for analysis for a
particular laboratory because of the many and varied organic analyses

available.   Private laboratories prefer to quote a price and time
required as determined on an individual basis rather than across-the-
board quotations for every parameter.


When attempting to identify and quantitate organic chemicals in water,
it is necessary to be familiar with some of the techniques used in
the identification of organic compounds in general.  Many physical
constants such as:


     A.  Melting points


     B.  Boiling points


     C.  Refractive indices


     D.  Density


     E.  Optical rotation


     F.  Molecular weights


can be determined in organic analyses.


Expensive instruments such as the NMR and the mass spectrometer can

also be valuable in organic identification.  However, in many cases
infrared spectrometry and chromatographic procedures prove adequate.


Gas-liquid chromatography is an analytical method for the separation
and identification of a mixture of volatile (usually organic)
components in a sample.  GLC can be extremely sensitive, but because
of this sensitivity it is often necessary to apply extensive clean-up
techniques.  Also, while GLC is ultra-sensitive and does an excellent
job of separation, it still lacks the specificity necessary for
positive identification.


Infrared analysis after separation and possible identification by GC
has found extensive use in water analysis.  Because no two compounds
will have identical spectra, an infrared spectrum of a pure compound

presents a definite method of identification.  The greatest use for
IR is to detect functional groups that are most commonly encountered
in organic compounds.
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    COLLECTION, PRESERVATION, AND ANALYSIS OF ORGANIC SAMPLES

                               by

                        Robert E. Reinert

    Fishery Biologist, Bureau of Sport Fisheries and Wildlife
                 Great Lakes Fishery Laboratory
                       Ann Arbor, Michigan
The most important aspect of monitoring fish kills is speed.  The
field biologist should make every effort to do three things as
quickly as possible:  get to the site of the fish kill, make the
collections, and get the samples to the laboratory.  If the fish
kill is in a river, samples should be collected above and below,
as well as at, the site of the kill.  At least several species
should be collected.  Comparisons can be made only between fish of
similar size classes of the same species.   If possible, 20 fish
of a particular species and size class should be collected at
each sampling site.  Only whole fish should be collected (unless
otherwise specified).   Representative bottom organisms also should
be collected.  Much the same procedure should be followed in
collecting samples in lakes.   Fish and other organisms should be
collected at the site of the kill and, if the lake is large enough
and the kill is localized, from several areas where they have not
been affected.  Again, every effort should be made to collect the
same species and sizes from the different locations.   Dead, dying,
and apparently healthy fish of each species collected from each
site should be packaged and labeled separately.

There is some disagreement among analysts as to the best way to
package fish.  Some types of plastic bags may contaminate samples
with plasticizers or PCB compounds.  Probably the safest procedure
is to wrap the samples in acetone-washed aluminum foil before
placing them in plastic bags.

All samples should be frozen,  if possible, or at least iced,
immediately after collection.   Analysis of a rotten fish is messy
at best, and because of the possibility of bacterial action, the
results of the analysis often are of little value.

Proper lines of communication between the field biologist and
laboratory personnel are extremely important in the monitoring of
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fish kills.  Each sample must be properly labeled to show species
and the locality, date, and method of collection.  Too often,
field people feel they are sending samples off into some unknown
void and laboratory personnel feel that that is where their samples
are coming from.  This feeling is eliminated if the biologist, in
addition to preparing a written report describing the conditions
of the fish kill, discusses the kill directly with the analyst.
Unfortunately, no two fish kills are ever exactly alike; consequently
the sampling and analytical problems differ with each situation.  A
field biologist who can communicate with the laboratory people will
better understand the analytical problems and will be better able
to collect samples that provide the most information to the analyst.
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                 INTERPRETATION AND USE OF DATA


                               by


                        John E. Matthews
         Aquatic Biologist, Manpower Development Branch
              Air and Water Programs Division, EPA
                          Ada, Oklahoma
The purpose of a fish kill investigation is to accumulate information
(evidence) that can be used to determine the exact cause of a fish
kill.  Often some of the necessary information is not available and
cannot be readily obtained.


Evidence concerning fish kills basically falls into three categories:


     1.  On-site evidence:  collected during the investigation at
         the scene as soon as possible after the kill.  Includes
         routine analysis of water samples.


     2.  Miscellaneous evidence:  collected by various means.


     3.  Laboratory evidence:  collected during subsequent laboratory
         investigation.  Includes field toxicity tests.  Lab
         investigations supplement field investigations.


The ability to diagnose the cause of a fish kill is highly dependent
on the manner in which samples are collected and the amount and
reliability of information recorded at the time of sample collection.


Although the investigation is a team effort and normally involves
numerous individuals in the collection and analysis of samples,
data should only be appraised by scientists or administrators
competent to do so.


"In general, the job of the scientist is to invent a story which
accounts for a set of observations and then to decide how likely
the story is."  Our starting point should be a set of conclusions,
and our purpose should be to find which of these conclusions are
most nearly correct and which are clearly not correct.  It is not
unusual for all preset  conclusions to prove to be incorrect.
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Use of chemical data from analysis of fish and water samples to
provide answers as to the cause of a fish kill involves not only a
familiarity with conditions under which samples were collected,
including visual observations of dead and dying fish as well as
water in the fish kill area, but also an awareness of basic principles
of fish toxicology.  There is no substitute for training, experience,
judgement and familiarity with toxicity and fish kill literature
in interpretation of data.

Even then data is not easy to interpret.  There is a tremendous
volume of toxicity literature, much of which is contradictory.
Determination of a concentration of a material that is equal to
or greater than that stated to be lethal to fish in some reference
is not conclusive evidence that the material caused the observed
fish kill.  Environmental variables such as temperature, pH,
calcium, DO, species and age of fish, and many others influence
the toxic actions of many materials, sometimes over one or two
orders of magnitude.  In addition most toxicity studies involve
exposures to continuous concentrations; whereas water samples may
reflect a concentration that existed for only a short time.

Problems encountered in interpreting and using chemical data are
usually inversely proportional to the complexity of the effluent
and the number of possible .sources.

There is also more to interpreting data than finding the presence
of a toxicant in fish samples.  The question is was there enough
of the toxicant present to kill the iish.  What constitutes enough
may vary from species to species and even from population to
population within a species.  Often it may be necessary to find
enough of the toxicant in certain tissue(s) of dead fish.  What
constitutes the right tissue must be determined for each individual
toxicant.  For example blood is the right tissue for several
pesticides; the gill for cadiniium; the gill and opercular bone
for zinc.  The Japanese have conducted considerable research in
the use of whole fish bodies and critical tissues for post mortem
identification of fish killed by toxicants.  Results of this
research is published in the Japanese Society of Scientific Fisheries
Bulletin and is referenced in the Annual Literature Review issue of
JWPC'F"

The key to using the critical tissue concept is collecting proper
samples.  Three kinds of samples should be collected for tissue
analysis:

     1.  Live unexposed fish.  (Control)

     2.  Dead fish.

     3.  Live exposed tish.
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The number collected of each is dependent on what you are looking
for.  Twenty (20) of each would be good but this may be impractical.
At least ten (10) of each is recommended.  Preferably all fish of
one species should be about the same size.

A lethal critical level is said to exist when the lowest level of
the toxicant found in certain tissues in fish killed by the toxicant
is higher than the highest level of the toxicant that can be found
in the same tissues in fish exposed to, but not killed, by the same
toxicant.

By using appropriate samples and analytical techniques, one can
attempt to determine if there appears tc be a critical level of the
suspected substance in any tissue for the sample fish.  This technique
possibly overcomes complications for hypersensitive fish due to other
stresses, resistant fish, and prior exposure.

This is probably the only kir.a of approach that should be used at
the present time to try tu prove- the v a use of a fish kill from
tissue analyses.  It cannot be used unless proper samples (kind
and numbers of fish) are collected and preserved during on-site
investigation.   Tissue analyses of only dead or moribund fish,
however, may be used to place possible suspected toxic agents at
the scene or to eliminate them all together.

The toxic agent and source cannot be positively identified unless
stream samples can be shown to be lethal.  After collection of a
sample assumed to contain the toxic substance or complexes causing
the kill, the sample must be proven toxit .  This can be done by
routine bioassay in the :najorit> of cases.  If concentrations are
of sufficient magnitude, a bioassay may not be necessary.

The relationship of other chemical conditions to the toxicity of
pure compounds has been studied intensively.  These contributions
are important but not too helpful in finding who, or what, killed
the fish.  In fact it is almost impossible to establish the cause
of a fish kill If positive identification and determination of the
conditions, chemicals, or complexes contributing to death are
required.

The addition of any chemical or waste that can be shown to change
a previously nontoxic complex to a toxic one should be considered
to cause the kill, even though the substances in the water act to
increase the toxicity greatly.  This approach now forms the basis
for the use of bioassays to establish the source of a kill when
additional supplementary evidence is not obtained.

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In summary I would say that the ideal situation  is  to  observe
conditions under which the fish are dying and actions  of dying
fish; to examine freshly dead fish; to make a reconnaissance of
the area for all possible sources; and from this information, come
up with a list of possible toxic agents.  Interpretation of
analytical data then, in some ways, becomes a process  of elimination.

Again the interpreter must be familar with the toxic actions of
all suspected agents.


One thing to keep in mind is that it should not  automatically be
assumed that subsequent fish kilJh in a waterway are caused by
the same reasons as preceding ones.  When investigating any fish
kill all possible causes should be kept in mind.


It is the responsibility of the scientist to interpret the data
in light of his experience, training, judgement  and all related
conditions and circums r.mrec-..   It will then be the  decision of a
lawyer to decide if th<-- evidence-  '.  -.tronj, enough for  litigation
action and a judge and/'->r  jurx  t  -  ''.riut.  if the  interpretation as
to what killed the LIsh is cor-''>-
                                ttn

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             FISH KILL INVESTIGATIONS AND TECHNIQUES


                               by


                           Ed Sorrels


         Arkansas Dept. of Pollution Control and Ecology



Fish kill investigative procedures involve:


     A.  Prep Lanning


     B.  Information to be gained from informant


     C.  Preparation at etfi.-e and laboratory


     D.  Area observation JTK!


     E.  Area sampling


     F.  Trace of causative agent


     G.  Documentation of events


     H.  Post-investigation procedures; bioassay, benthos

         moni toring


Speed of response has been stressed by a number of participants

in this Seminar.  Preplanning is vital for speedy response  to a
report of a fish kill.  It requires provision of a prompt fish
kill reporting network, trained personnel who will make the in-
vestigation, and the necessary equipment for thorough procedures
and facilities for all pertinent analyses.


The Arkansas Game and Fish Commission has 125 enforcement officers

stationed over the State and 12 biologists in district offices, all
in radio contact with the central office in Little Rock.  In turn,
this is linked by telephone through the State governmental  exchange
to the Department of Pollution Control and Ecology.  A specific

section of the Department iias the responsibility for investigating

pollutional-caused fish kills.  The Ecology Section of the Water
Division of the Department comprises 5 ecologists and 2 technicians

who are involved in routine biological monitoring, annual stream
                                87

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basin water quality  surveys  and periodic special surveys; however,
1 or more  is  ready  to  drop  current activity and proceed with a fish
kill investigation  at  instant  notice.   In addition, the Department
maintains  9 field inspectors and 7 district offices, as well as 2
mobile laboratory units  located for current water basin surveys.
Ordinarily any  area  of the  State can he .reached by a trained
investigative team within
a fish kill.
                             he^irs  of first receipt of report of
                           -e t
                            - e-t
The Department Chemistry  Serti
routine water sample s  01
stations of the  Inters!,,
furthermore operate  fro-
watei basin survey:-.   Th
chemical anal vs is  de^-ri
of Water and Was tew at or'1
extrae'tion and gas thro1'-
and tissue sarirlos.
As much pertinent  inform
the observer reporting  t
mation form is appended
should arrange to  meet  t
the investigator to  the
directions to the  fish  :-
or was first observed,  u
if not, when it stopped,
to miles of stream af fee-
percentage of th«  e1-. tira
the species and si/<   * JH
per yard of hank?  what
as to cause?  u'hat rece:>
basin, such a- crop  du: t
overs'"'  What has boor.  Hi
there been heavy rainf.i'
period of cloudy ovettas
sudden temperature chain'
upon the proMem.  U'hai
land uses—whether it  di
population dens it\?   Ha--
is there a periodiciiv  -
conditions or time' uf  ••
                                  i-; i ont inually engaged in analyzing
                                  iroughout the year from over 70
                                   S;uer Ounlitv Network.  Chemists
                                   ilf laboratories during specific
                                   is capable of performing any
                                  ;  indard  Methods for the Examination
                                   rion  also has facilities for
                                   • •'  pi stie;j€-s in water, sediment
                                         e- should be obtained from
                                          copy of a suggested infor-
                                         Where possible the investigator
                            reporting individual and have him accompany
                         i ish  kill  area.   Bv all means obtain precise
                         ill area.   Determine when the kill first began
                         hetiier  or  not  the kill is ce>ntinuing at present;
                           Get  ">n estimate of the extent of the kill as
                         ted or  size of pond or lake covered.  What
                         f. ;i fi^-h r opn l a i i on was affected?  What are
                         •f;-. ^  '• :*  -~ved '   • -t i mated number of dead fish
                         is.  ihe   r -' n i ui . :  the reporting individual(s)
                         t  activities have taken place in the drainage
                         log.  MidusLrial  spills, start-ups or change-
                         i  wfiiiu'r  pattern preceding the kill?  Has
                         I9  When?   Duration?  Or a long extended
                         t?  ilighwind'-, wind direction, temperature,
                         es, i-'tc.;  aJ 1  of  these questions may bear
                         is  tho  nature  of  t!ie watershed and predominant
                         -ins  municipal areas or areas of considerable
                         '-'  ••, i ;-• i Lai  fi ;h kills occurred before?  If so,
                         ;  an  association  with other events, weather
                         'ir
On gaining
of trii  kill
                 nil
               the
                         ion,  the  investigator pinpoints the location
                         .-I'-taii-,"-: needed  maps  of the area from files,

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determines  possible sources  of-  roIiutM-r.
non-routine sampling ana anals -is rue.  he
pole-preserving (creosoting)  j 1 an t m;-v hi.
wastes phenolic-- (creso1' ,).  pi P t -i-di 11 r-oi-i
oily  carrier.   An electroplating  jncj'str'.
agent.   In  water, hydrogen cv i.iide is  tht
a cyanide  concentration which  is  Limocuoi
pH 6.  Preservation of cvanides,  in .jcklit
by adding  20 ml of 1 N NiOH  poi  lire-  --,f
in the discharge of industrial  v,«s!e-  rr
from  ice plants, or from scour ir-   ir.d
"ammonia water'1 is usee.  Pr< .'• >v  •' ;> ;% •
icing, is  accomplished h\ fidu: -,g  •'-(' i.if. i;
The investigator notifies tin  , demist '-\
estimated numl-et of sairrlo.-  jt;,;
estimated  time  of ntum \ ,.   ;1   '
that  may be required.
                                           and decides  whether any
                                           required.   For example, a
                                            expected  to have in its
                                           enol.  and  possibly a complex
                                            uses  cyanide as a complexing
                                            major toxic factor, so that
                                           s at pH  8  may be lethal at
                                          tion to icing, is accomplished
                                           sample.  Ammonia may be found
                                           m chemical or gas plants,
                                           an ing  operations where
                                             n:p-'or i a ,  in addition to
                                           t I-} re:  liter of sample.
                                           ahoratorv  operator of the
                                           analvses to  he expected, the
                                           \, and of  an^  special analvses
A suggested  c. o> k.li , r   •• <.
is appended.   A good port >e",
in portable  cases designed  L
for the  fish  '-.ill i uve^r i gat
under way  read i Iv li\- ga^.'u
sampling contairer  -.

On c e ir;  the  f i. e Id,  d i s c •• n
fish kill  invest igat i(.n .
fish ire still  dyni-  n1".-'
and analyze  the re'i lis'i-
can be deterr.i nod .   Kt
inves t i gciti o!',   0m  ,-,<;.
to "tell the  st,>rv' , ',o  -
happened wfuh  ,:n ', :
collected  shoill  p  •••v-
(It may  be t'ut ^or- • •. .•
Some rough  stream .'. i  i.'i
physical  surv-. \  in- ^ i \'
questions art  ; t ^e iteil
and tracing ' >u  ' .ci-  it

Are there do^d  risi- '
(In the case  where -.-,n
apparent  on invosti.-i.it
and dredge  the  aot t v
                                                t:>,'  "lake  tne investigation
                                                v equipment can be kept
                                 e..   rs ._\;e,  r r

                                 tie  purpose in  the  vehicle reserved
                                 ^  s,-; i-;iat  the investigator can get
                             ins-'  uu  an ir" 'o id or  so of  enuipment and
                                        L on is the  kevnote of a thorough
                                       >._, t [ jatt-r Arrives  in the area while
                                        H'».-^s'irv observations, collect
                                                 the
                                                           of the kill
                                              ^ t. -  ' ripiariJx  on the field
                                              11.111  laboratory analysis
                                              .'.-,; ,-j L  basis  of what has
                                              ion rvide ,  e-u-h sample
                                               , a( stioii,  \'es or no.
                                              we red  directly.)

                                               iiustrdte  the overall
                                               >r areas  wnert.: these
                                              lo  -'^urces  of  pollution
                                     riv; Hs'-?  Are  there  live fish?
                                     rvi'f.1 do.id fish,  but  none are
                                     o.src1'.   Clieck  the  bank for remains
                                     >. ')SL rve tue water  and the dying

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or dead fish.  Note your observations.  Penciled notes in a water-
resistant field logbook are suggested.  Is the color of the water
green, brown, reddish, or black and septic?  Does it have an odor?
Try to characterize the odor; e.g., rotten eggs, cucumbers, sewage,
etc.  Is the water clear, even though colored, or is it turbid?  Is
the turbidity from grossly visible particles or does it seem to be
from fine particles?  Does the bottom have  a thick layer of sediment?
Are there conspicuous growths of attached green algae, or patches of
light brown sewage fungus (Sphaerotilus)?  Is there associated with
the kill some other physical presence that can be traced, such as oil
material that creates a skim or "rainbow" and marks vegetation at the
water's edge?  The banks and vegetation can indicate some chronology
of the water level.  A rise that has passed generally marks the
vegetation to the highest level obtained.  Submerged green leaves
indicate higher than normal levels.  Some agents actually kill
susceptible vegetation reached by a rise.

Note the behavior of dying fish.  Are they nervous, skittish,
convulsive?  Are they "piping"—gasping at the surface—, or
listless?  Note the appearance of dying fish, their gills, fins,
scales, abdomen.  Look for discoloration, hemorrhages, fraying,
sores or cysts, whitish slime, swollen abdomens.  Is there a
predominance as to size of fish and species affected or is the
distribution random?  Tabulate size and species distribution of
one or more  collections of affected fish with appropriate notes
as to representativeness, etc.  A finding that only the smaller
fish died or  that  large  fish  died but small fish survived is
important in distinguishing between oxygen depletion and toxic
causes.  The key difference is gill surface ratio  to total body
mass.  Small fish, being relatively more rapid in absorbing water
solutes, withstand marginal oxygen depletion but succumb first
to toxic materials.  Observation that animals other than fish are
killed—snakes, turtles, etc.—will certainly indicate that an
extremely toxic factor other than dissolved oxygen depletion is
involved.  The observations made according to this section should
enable the investigator to estimate whether the kill is an acute
situation or a continuing chronic problem.

Record the temperature of the water.  If very deep, check it at
several depths.  Test the dissolved oxygen, always!  On a small
stream five feet deep or less collect the sample at mid-depth,
midstream.  On deeper streams collect the samples at 0.2- and
0.8- depth.   On broad streams collect these samples at quarter-
points also.   The absolute minimum of samples to be taken are:
                                90

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(1) In at least one point downstream, preferably where fish are
actually dying; or where kill was heaviest, if over.   (2) In source
effluent, if any.  (3) Above kill area or suspected source.  On ponds
and lakes sampling on several transects may be required; across,
above and below the fish kill area.  Sample in source  effluent, if
any.  Also sample on windward side and on leeward side.

If the dissolved oxygen is adequate at all depths and  all areas,
B.O.D. samples are omitted.  If oxygen deficit is a possible factor,
a B.O.D. sample from the area is to be collected.  If  there is a
heavy growth of algae, a plankton sample also is collected.  If there
appears to be an algae dit—off with a surface scum and/or clear
brownish water, a skimming rf the surface also should  be collected.

If a source of organic Tollution is a factor, a grab sample and
B.O.D. sample from the effluent must he collected.  If there is
a suspected source of nnmioipal waste, Cannery waste,  or feedlot
waste, a routine bacteriological sample (total coliform, fecal
coliform, fecal streptococcus) from the kill area may  also help
in establishing the causal connection.

Measure the pH.  The appearance of the water and nature of suspected
sources of pollution are clues to whether the reaction may be a factor.

Each sample collected should be labeled immediately with the name of
the stream or lake, the sample point number by noted map or diagram,
the date and time of collection, the analysis required, the bottle
numbers of 0.0. and P.O.D. samp Its collected at the same point and
time, the air and water temperature in degrees centigrade, and the
initials of the i n\,ijst i e,nt or -   S.imrle;- should be sealed and iced
inlinedi ately ,

Collect on H  distressed or civ ing fish for lab examination and/or
blood and tissue analvsis.  Collect at least one individual preferably
two, of each, species affected.  Place in plastic bag,  tag with date,
time, stream and location, condition of fish when collected, as well
as suspected causative factor.  ice, and freeze as soon as possible.
Take blood samples if pesticides or toxic chemicals are suspected.
Cut through the hodv of the fish ahead nf the caudal fin and "milk"
blood from the dorsal vein iind aorti into a prepared vial.  Collect
at least 5 ml from o uh species.

Make an estimate of the sizt and extent of the fish kill; the species,
sizes, numbers and weights, and the miles of stream or acres of lake
affected.  In Arkansas the State ;"ame and Fish Commission will make an
extensive ana thorough evaluation mi large kills.

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In the field  the  investigator  is working  against time.   His critical
samples should be back  in  the  laboratory  for analysis within 24 hours
of collection.  This  is  usually readily accomplished with some overtime,
perhaps,  Even if he  spends  the entire day in the field he can have
his critical  samples  in  the  laboratory the next morning.

Answers to  the questions indicated  in  the paragraphs on observation
and sampling, at  the  key points in  his physical survey, will enable
the investigator  to trace  the  causative agent to its source, ruling
out other possibilities  in  t'irn.  .iud i c iousl v-scened photographs can
aid in documentation  ,il  rtn.  i i rcunu- t jnce-, of the kill.   Once the
agent is traced to its  sount ,  tht  investigator may require entrance
and inspection of the source.- \ur-ti-  facilities and storage facilities.
He should inform  plant  :tidnaget?cn t t. nat ho. is investigating the specific
kill, that  'no has trcut-^i the agent  t_-->  the plant, arid request they
a (.company li.'tr. on  the  Gcsiret  inspection.   Tn Arkansas,  such entrance
for the purpose ol y^ilutioi, : -iv<"-; t i ?,> tion is specifically provided
f'1; in the  St.tte  i\'l]ut'"n  (ontro]  j-n:s.

The investigator  may  find  it appropriate  to take statements, signed
and witnessed, from observers  of events pertinent to the investigation.

In the case of complex  and  poorly-defined wastes, documentation of
toxic effects and levels of  toxicity may  be well-served by bioassay
procedures.  These procedures  should always be carefully planned,
defined and recorded  along  the  lines  of  standard scientific methods.
The benthic community of a  body of water  is subject to the effects
of the materials  that pass  through.  A profile of the number and types
of organisms  according  to  their sensitivity will serve  to document
the unaffected portion,  the  zone of degradation and the zone of
recovery or of marginal effects.  Iji_s_ij;u monitoring can be accomplished
by diverse means  such as caged fish, rock basket benthos samplers and
floating anchored periphyton samplers,

Remember that information  flows both wavs:   Water Quality Criteria/
Field Determinations.  A well-documented  fish kill investigation with
levels of the toxic agent  established  and definition of conditions
obtaining prior to and during  the fish kill will also serve as a
bench mark  for future work.

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                            APPENDIX


1.  Standard Initial Information Form for Fish Kills.

2.  Checklist for fish kill investigation equipment.

3,  General diagram of stream and tributaries to emphasize
    importance of checking and ruling out possible physical
    sources as the investigator moves upstream in tracing the
    causative agent of any fish kill.

4.  Illustration of specific fish kill.   Heavy rainstorm
    washed-over inadequately free-boarded holding pond causing
    a total fish kill in some thirty-five miles of Little River.
    Total poundage 3^4,000; total value $46,382.  Previous
    bioassays had shown that even treated waste from this plant
    was highly toxic  (TLn/,^ = (3.05% dilution).  Forms of life
    other than fish were killed, including some vegetation.
    Water quality par meters (D.O., nH,  etc.) were normal except
    for turbidity.  Agent  w;>s iriced i o source by dead fish
    (where anv alive preceding kill), bv phenol content of samples
    and by 'narking of water . banks and bank vegetation by the
    oily carrier.

    Some three weeks later a partial kill occurred in Old River
    Lake.  Sources ut other toxic 1actors were ruled out, a
    residual of unemuisified material from the original spill
    was demonstrated, and it was shown that movement of some
    of the residue had occurred with reversing of normal flow
    in channels with dispersion of some of the material over a
    portion of" the lat-e bv wind ana rain.

5.  Illustration of specific fish kill.   A number of fish were
    killed on east side of lake.  Wildlife officer found empty
    herbicide cans at the water's edge near the kill area.
    Other findings were consistent with conclusion of indirect
    kill from oxygen depletion due tc algae kill.

6.  Illustration of .specifi.-: fish kill.   Trace of causative agent
    showed that acid i\ino waste had been pumped from a reopened
    pit some fifteen stream miles from the kill area.  Spraying
    of power-line right-of-way was ruled out as a significant
    factor.

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                                        flfy  "lj,L

                                    ir~T]A  v..ici  '• 'hi'
          1.   Commission inf^rrr.at i "n -'htained  fror.i:

               Name
               Address _____ _________ __ ________ Directions to home or office -



I        2.   Initial .-bs,   Ocntin.ii nr at present.   (yes or no) if no,  when
                             stc r P' od
•        5.  Extent  of rill  (A)   Area covered -  (miles c:' stream or  size
                                     i  }, -.r,d or lake) .
          6.  Apt r'-xi:-atc arc-,.:t oA  fish affected  -  ( ".  of estimated populat-
              ion of area 9 f: roted ) .
              B.
I


I


I              	            	

              I____	.	:	___,	--..- —~.		  	 „	   	      	               ______
          S.   I-e-;«r.t,  ,ji;.-it;-   in area  (dujtinr, etc. to  include recent
              \j>: •;* '  n ••

•        9.   Pcs^'Aniv  _  ,: •'*-  -  ' - ;~l-,t'-'  in area.   (Known  to  reporting
•                   -
         10.   Anrri ^r: - JIL -r^ S',aten:.i:t '.ni'  cl^jr-cr and Commission personnel
•            "I"'  -K  -:/. _______________________ ..... _________ ^ ___
                                              94

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         CHECKLIST FOR FISH KILL INVESTIGATION EQUIPMENT
D.O. and B.O.D. bottles; alkaline azide and manganous sulfate
solutions; A.P.H.A. sampler.


Liter-size plastic grab bottles; 2- or 3-liter pesticide grab
bottles with foil-lined caps; benthos jars, formalin, string tags,
pencils.  Thermometer.


Ice chest and ice,


pH meter, kit or fresh pH paper; portable D.O. probe; Hach Field
Laboratory Kit.


Dip net, plastic bags for fish, dissecting kit.  Vials for blood
samples.


Needed maps, county and quadrangle.


Dredge, spade and screen.


Vehicle, boat and outboard motor.


Camera and film.


Paper towels, distilled water; lantern; appropriate clothing and
boots.  Flash light and batteries.
                                95

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            PHYSICAL
             PHYSICAL
 WIND
 #4//v
S70&M
                                   A/0&MAL D.O.
                                    FNR FISHING
                                        YBLLOW
                           96

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                        A/O&MAL FISH AND
                         AQUATIC LIFE
T&LATMENT PLtW7_

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PHYSICI\L  SUJ3V£Y   -5
                              WAT£K CLEAI?
                             GOOD AQUATIC
                               LIFE
                             £>£AD PISH
                             De.i\t>
                             pH 5 &Q6
                      LIVE.  fISU
                 K//\7£/P CLEfiK GREEN
                 LIV£  PRODUCTIVE POOL
                         PREVIOUS F/SH KILLS

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                                      KIL1, NUMBER ESTIMATES
I

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I                                             ijy
                                           Byron Moser
•                          Oklahoma  Dept.  of Wildlife Conservation

I

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•            The  foLlovvTing report  prepared bv the Pollution Committee, Southern
              Division, American  FishorJes  Sr>i:ietv,  1971,  was used as a background
I
              for the presentation  bv  Nr,

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                BASIC PROCEDURES

          FOR INVESTIGATING FISH KILLS

                   IN STREAMS

                   PREPARED BY

   THE POLLUTION COMMITTEE, SOUTHERN DIVISION

           AMERICAN FISHERIES SOCIETY

                      1971

           Billy J. Grantham, Chairman


                COMMITTEE MEMBERS
Sam Spencer
Jim Collins
Joe Blanchard
John Frey
Peter Pfeiffer
Bobby Walker
Benjamin Florence
Roger Hogan
Byron Moser
Miller G. White III
Daniel M. Sherry
Kenneth C. Jurgens
Eugene W. Surber
Howard Zeller
Alabama
Arkansas
Florida
Georgia
Kentucky
Louisiana
Maryland
North Carolina
Oklahoma
South Carolina
Tennessee
Texas
Virginia
Federal Water
Quality Administration
                       100

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                BASIC PROCEDURES

          FOR INVESTIGATING FISH KILLS

                   IN STREAMS

                   PREPARED BY

   THE POLLUTION COMMITTEE, SOUTHERN DIVISION

           AMERICAN FISHERIES SOCIETY

                      1971

           Billy J. Grantham, Chairman


                COMMITTEE MEMBERS
Sam Spencer
Jim Collins
Joe Blanchard
John Frey
Peter Pfeiffer
Bobby Walker
Benjamin Florence
Roger Hogan
Byron Moser
Miller G. White III
Daniel M. Sherry
Kenneth C. Jurgens
Eugene W. Surber
Howard Zeller
Alabama
Arkansas
Florida
Georgia
Kentucky
Louisiana
Maryland
North Carolina
Oklahoma
South Carolina
Tennessee
Texas
Virginia
Federal Water
Quality Administration
                       100

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                BASIC PROCEDURES

          FOR INVESTIGATING FISH KILLS

                   IN STREAMS

                   PREPARED BY

   THE POLLUTION COMMITTEE, SOUTHERN DIVISION

           AMERICAN FISHERIES SOCIETY

                      1971

           Billy J. Grantham, Chairman


                COMMITTEE MEMBERS
Sam Spencer
Jim Collins
Joe Blanchard
John Frey
Peter Pfeiffer
Bobby Walker
Benjamin Florence
Roger Hogan
Byron Moser
Miller G. White III
Daniel M. Sherry
Kenneth C. Jurgens
Eugene W. Surber
Howard Zeller
Alabama
Arkansas
Florida
Georgia
Kentucky
Louisiana
Maryland
North Carolina
Oklahoma
South Carolina
Tennessee
Texas
Virginia
Federal Water
Quality Administration
                       100

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                BASIC PROCEDURES

          FOR INVESTIGATING FISH KILLS

                   IN STREAMS

                   PREPARED BY

   THE POLLUTION COMMITTEE, SOUTHERN DIVISION

           AMERICAN FISHERIES SOCIETY

                      1971

           Billy J. Grantham, Chairman


                COMMITTEE MEMBERS
Sam Spencer
Jim Collins
Joe Blanchard
John Frey
Peter Pfeiffer
Bobby Walker
Benjamin Florence
Roger Hogan
Byron Moser
Miller G. White III
Daniel M. Sherry
Kenneth C. Jurgens
Eugene W. Surber
Howard Zeller
Alabama
Arkansas
Florida
Georgia
Kentucky
Louisiana
Maryland
North Carolina
Oklahoma
South Carolina
Tennessee
Texas
Virginia
Federal Water
Quality Administration
                       100

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                          INTRODUCTION
The Pollution Committee, Southern Division, American Fisheries
Society recognized that the most paramount problem following
the establishment of the monetary values of fish by the 1970
pollution committee was to establish guidelines and procedures
for determining the size of fish kills.  Only two of the states
in the Southern Division have written standard procedures.  The
methods employed by other states varied considerably.


It was also recognized that in all cases the primary responsibility
of the Conservation Agency was to delineate the kill by species
and magnitude and not to identify the exact toxicant.  This latter
responsibility is in all cases designated to the Pollution Control
Agency.  Although the Conservation Agency is not charged with
identification of the toxicant every effort is made to do this
to offer supportive and corroborative evidence to the Pollution
Control Agency.  Furthermore, detailed field notes, photographs,
etc., are made to lend added support.  In addition to the types
of data acquisition mentioned above, all of the Conservation
Agencies examine the fish for disease conditions and if necessary
seek the services of disease specialists if there is a doubt.
                                101

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     BASIC PROCEDURES FOR INVESTIGATING FISH KILLS IN STREAMS
Before any plans can be formulated for making a count of dead fish,
it will be necessary to determine the miles involved in the kill.
This area should then be delineated on topographic or other suitable
maps.  The most desirable method for determining the number of dead
fish is for all to be counted, but this is often impossible.  If
the investigator deems it necessary to make only a partial count,
this must be done in a manner that will produce unbiased results.
The committee felt that an appropriate procedure would be as
follows:


     Make 100 yard counts every one half mile starting with
     the first dead fish observed.  The additional counts
     should then be made at each half mile interval throughout
     the region of the kill.


Each count will consist of the following:


     1.  Identify, count, and determine inch groups of all fish
         in each 100 yard segment.


     2.  Calculate the total number of fish killed by species in
         each count area.


     3.  Multiply the tola] number of each species by 8.8 (the
         number of 100 yard segments per 1/2 mile).


     4.  The answer obtained in item 3 represents the number of
         each species killed.


In order to facilitate use of this method an example is included
below:
1st 100 yards
Species
Blue gill
"
"
"
"
"
11
it
Number
140
120
60
30
25
30
10
5
Inch group
1
2
3
4
5
6
7
8
                                        420


                               102

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2nd 100 yards
3rd 100 yards
4th 100 yards
Species Number
Blue gill 100
80
40
20
15
15
5
	 5
280
Blue gill 40
30
20
15
25
20
10
_JL
165
Blue gill 0
10
5
10
15
5
0
0
Inch group
1
2
3
4
5
6
7
8

1
2
3
4
5
6
7
8

1
2
3
4
5
6
7
8
                                         45
5th 100 yards       End of Kill
Total fish counted  910
Number of miles     2^
910 X 8.8 = 8,008 Blue gills killed
                               103

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Counts such as those illustrated should be made for each species
and the total number of fish killed calculated.  Since the kill
will be assessed by use of the monetary values, it is necessary
to have a breakdown to the various inch group catagories.


In the event of excessive large kills over many stream miles the
investigator may deem it necessary to make counts at one mile
intervals.
                                104

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                         VALUE OF FISHES

                               by

                         Sam L. Spencer

           Chief of Fisheries, Game and Fish Division
    Alabama Department of Conservation and Natural Resources
If you plan to go to court to recover compensatory damages for
dead fish, you must have a value to place on the fish that were
killed.  Prior to 1970, only four of fourteen Southern Division
states of the American Fisheries Society had a list of monetary
values for their fish.  These values varied considerably among
these four states.  For example, a 12" bass in the Tennessee River
could have had a different value depending upon whether it was
killed in the Tennessee, Alabama, Mississippi or Kentucky portion
of the river.  What would have happened if a fish kill occurred
near the state line and fish were killed in two or more states
and these states used different sets of fish values?

In 1970, the Southern Division, American Fisheries Society,
formed a Pollution Committee with one member from each of thirteen
states and one member from the Federal Water Quality Administration.
The states included were Alabama, Arkansas, Florida, Georgia,
Kentucky, Louisiana, Maryland, Mississippi, Oklahoma, South Carolina,
Tennessee, Texas and Virginia.  The first task undertaken by this
committee was the development of a set of fish values to be used
as a guideline.  At the end of 1970, the committee published their
"1970 Monetary Values of Fish".  I would like to read a few lines
from the Introdcution of this booklet which may help explain how
the values were derived.

"Every fish should have a monetary value placed on it.  The purpose
of a monetary value list for fish is for a fair and reasonable
assessment of a fishes' value for collecting compensation for the
destruction and/or loss of fish due to water pollution.  The
establishment of a list of monetary values for the many species of
fish was a complex task which required the experience and judgment
of the various members of the Pollution Committee.  Several hundred
commercial fish hatcheries were contacted to determine price and
availability of any and all sizes and species of fresh water fish.
Some problems were immediately encountered such as the non-availability
                                 105

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of certain sizes and species.  Another problem was the fact that
a "native" or wild fish specimen is usually considered a more
valuable resource than a hatchery-reared specimen.  Consideration
was given to establishing values of fish based upon what it is
worth to a fisherman to catch a given species and size of fish.
The Committee did not feel this approach was practical or valid
for the several following reasons:

     (1)  Many species are important in the ecological balance,
          yet are seldom pursued or captured by anglers.  Under
          this type evaluation such fish would have no value.

     (2)  The value for a given fish would vary greatly according
          to the skill, experiences, likes and dislikes of
          individual anglers.

     (3)  In many cases only a small percentage of sport fish are
          ever harvested by anglers.  Only this percentage could
          be considered to have a value.  For example, if only
          10 percent of the bass are normally caught, then the
          remaining 90 percent that die of other causes could
          have no legal value assigned to them.

"Therefore, where possible, the best assessment of this value was
based upon what it would actually cost to purchase or replace or
restock given fish specimens.  This was determined from the price
lists received from the many hatcheries contacted.  Certain other
fish, such as the sturgeons, were considered to be a non-replaceable
resource.  Thus, a price commensurate with the loss of this rather
unique fish was assigned.  Where hatchery produced replacement stock
was not available, the Committee had to judge a replacement value
on the role that this species plays in the ecological balance of
a body of water and not on what it would cost to rear and stock a
given number of specimens.  The Committee's judgment of these values
was based upon approximately 200 years total experience and training
in the fisheries profession.

"Admittedly, at times, certain fish may be purchased live at lower
prices.  However, this list attempts to take into consideration
the availability and nonavailability of numbers of some species.
Also, these values do not reflect the almost immediate loss of
some fish that occurs when hatchery-reared fish are stocked into
public waters.
                               106

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"We recognized that the laws of several states did not allow the
assessment values for fish killed by pollution.  It is hoped that
this report and list may be helpful in changing this.  We also
recognized that certain Southern Division states already had or
were developing their own list of values.  The adoption of this
list by all Southern Division states will present a more united
front to the pollution abatement effort which must be a goal of
highest priority of all fishery scientists."

As pointed out, some values may be considered arbitrary; however,
this will always be the case.  There is no formula you can use to
put in the fishes' scientific name, add some magic factor and out
will come an exact value that cannot be questioned.  We have to
accept the fact that there is no precise  way to determine the value
of a sturgeon, madtom or darter and there will always be regional
differences in how a particular fish is valued.

For these reasons, the list prepared by the Pollution Committee
was  designed to be used as a guide; however, it was hope of the
Committee that all Southern Division states would individually
adopt the list, thus giving it more legal strength.  At present,
a majority of the Southern Division states and several other
states have officially adopted the list.  We have been informed
that the values have been used successfully in several law suits.
We have found that when opposing lawyers ask how the values were
determined, if you show him this booklet that was prepared by 14
professional fishery workers with over 200 years total experience,
he usually has no further questions on the values.

The fish are listed by species and most species in the South  are
covered.  The fish and values are listed by each inch group up to
the length at which the species reaches one pound; then the values
are listed  per pound.

The usefulness of such a list depends upon how the law in your
state is written.  For example, our law in Alabama states that
the values are to be determined by the Alabama Department of
Conservation.  We have adopted the Southern Division list and the
Alabama Water Improvement Commission (AWIC) has officially adopted
the list and approved its use.

Another different situation is the New York law which states that
each fish killed may be valued at $10.00 each.  Therefore, New York
is not too interested in our list, and I don't blame them.
                                107

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The Pollution Abatement Committee of the American Fisheries Society
hopes to prepare a guide for all fifty states next year.  If such
a list could be prepared and adopted by the Society, it would give
considerable strength to fish values in court.

We heard mention yesterday of "significant" fish kills as perhaps
those involving game fish.  The Committee's feeling on this is summed
up by the first sentence in the introduction:  "Every fish should
have a monetary value placed on it".  Many of us feel that today's
rough fish may be tomorrow's game fish.  We see this happening in
Alabama now.  There is a move underway by sportsmen to have catfish
designated as game fish.  Catfish are the fourth most sought after
species by Alabama sport fishermen, yet, it is classified as a
rough fish.  In addition, many of our fishermen are enjoying
fishing for carp because they can be attracted in large numbers
and caught on sporting tackle when other fish won't cooperate.
We even sponsored our first carp fishing rodeo in Alabama this
year, and it was very successful.  We haven't learned how to enjoy
eating them yet, but they are fun to catch and we have a very good
supply.

If you find you must prepare a monetary list of values for your
state, I would suggest that you gather together as much information
on fish prices as you can, get as much good professional help as
you can, then be prepared to make a lot of tough decisions.  Many
hours can be spent arguing about a few cents on the value of a fish.
You must accept the fact that there are very few absolute values
that cannot be questioned.

In court you may possibly encounter an opposing lawyer that wants
to question your values; however, they usually prefer to question
the number of fish you reported.  The lawyer may state, for example,
that menhaden only bring $30.00 per ton and you are asking $.15 per
pound or $300.00 per ton.  You can counter this with the fact that
you are referring to the value of the live fish and he is talking
about fish his client killed.  Dead fish floating on the surface of
a public stream or lake are a liability, not an asset.

As I mentioned previously, usually an approved published list will
prevent further discussion of such values.  However, if the opposing
lawyer continues to argue the point, I would suggest that his client
be given the choice of either paying your values which are based
upon expert opinion or replace the fish with the exact number, size
and species killed and somehow compensate fishermen for lost fishing
opportunities that occurred as a result of the pollution.  If the
defendant has killed a few sturgeon, or striped bass, or pirate
perches that are not available from commercial hatcheries, the
lawyer may decide your values weren't so unreasonable after all.

                                108

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                     OTHER ECONOMIC DAMAGES


                               by


                         Gene F. Forbes


      Fishery Biologist, Tulsa Office of River Basin Studies
              Bureau of Sport Fisheries and Wildlife
A fish kill is indicative of much more than a loss to a single
resource.  It is an indication that the integrity of our water
has been violated and that all values dependent upon its quality
have been reduced.


When appraising environmental damage the total effect on all the
resources in question should be considered.  In order to evaluate
this damage the value of the "potential uses" must be known.


The potential uses of a resource are both consumptive and nonconsump-
tive with tangible and intangible values.


Consumptive uses such as irrigation, municipal and industrial waters
have tangible values.  The water quality required, the volume needed,
and the value of these waters for both the present and future genera-
tions are relatively easy to determine.


Nonconsumptive uses have both tangible and intangible uses and
values.  Tangible uses include such things as fishing, hunting,
swimming, boating, and many other outdoor oriented activities.
The magnitude of these are determined in man-days use.  These may
be calculated by actual counts, by sampling, or by other various
methods of estimation.


The value of the tangible uses may be determined by ascertaining
the price an outdoor recreationist would be willing to pay, the
net profit which a private operator would make, or by the value
to the local or state economy.


The quality of the experience and the other psychological reactions
which are an inherent part of outdoor activities have intangible
values.  Other intangible values include such things as protection
of wild and primitive areas, areas of unequaled beauty, scenic,
historical and scientific interest, and the preservation of rare
and endangered species and their habitat.
                               109

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Intangible values are much more difficult to define monetarily.
Those associated with active participation should be reflected
in the value assigned per man-day use.  Other monetary values
might be estimated using the cost necessary to protect the
intangibles in question, or by using the single-purpose alternative
method.  In this latter method the cost of developing a similar
area to replace the intangibles "in kind" is equal to the value
of the area in question.

A separate evaluation should be made for each area with the significant
physical, biological, or geographical differences defined.  This
inventory should include the amount, type, and quality of the waters
and the demand for  the resources available with these waters.

With this basic data on supply and demand the needs of the area may
be determined.  From these needs the potential uses may be derived.

The potential uses may be based on todays needs, but I suggest that
they be evaluated on projected needs.  A projection may be made
using population projections in conjunction with long-range programs
and anticipated public demands.

There are numerous advantages to using this method but I believe
the following are primary:

     1.  Long-range projections require updating as new data become
         available but it is not necessary to conduct an annual
         survey or census.

     2.  This method will standardize evaluations from year to year
         and insure consideration of all applicable uses.

     3.  When repeated or continuous pollution damage occurs it is
         much easier to evaluate the total effect.

     4.  In addition to using this information to determine pollution
         damage it may be used to establish long-range goals and act
         as a guideline in evaluating current programs.

Once the values of the water and its related resources have been
established the cost of a loss due to pollution may be equated to
the value of the potential uses that were lost or reduced.

Thus in a single case of pollution with a short term effect the value
of the loss would be minor.  But, if the pollution was cronic or of
a lasting nature the cost to the resource would be considerable.
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Any form of damage assessment or fines will be a deterrent to
violation of the water quality, but this is only treating a
symptom and not the cause.


People must be made aware that immediate economic gain at the
expense of good environmental quality is not good business.  In
the long run continued pollution will cost the nation more in
both tangible and intangible values than the cost of prevention.
                                Ill

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               THE FISH KILL INVESTIGATION REPORT

                               by

                        John E. Matthews

         Aquatic Biologist, Manpower Development Branch
              Air and Water Programs Division, EPA
                          Ada, Oklahoma
Every fish kill investigation that is worth making is worth reporting.
Even the "wild goose chase" should be documented in some manner for
possible future reference.

The report should be a systematic recording of organized thought to
convey the fact findings of the investigation accurately, completely,
and understandably to the reader.  The final report should be such
that it can be introduced as evidence into court and withstand
critical scrutiny and cross-examination.

The final fish kill investigation report is largely dependent upon
the investigator's compilation of a thorough and complete set of
notes about the contacts made, visual observations, and field and
laboratory analyses conducted.  All field notes, laboratory notes,
and data forms must be maintained in the agency files until there
is assurance that they will not be needed further in court actions
or elsewhere.

The report should be prepared as soon as possible after completion
of the field work, while the details are still fresh in the mind of
the report writer.  Details of the situation start to fade from
the memory in a relatively short time regardless of how complete
field notes may be.

Reports of findings in fish kill investigations should be made only
by competent personnel (biologists) thoroughly qualified to judge
the nature and completeness of the data collected.  The writer
should have participated in the field work ensuring familiarity
with the waterway, sampling locations, water sources, and details
of the field operation.   This knowledge is essential for the most
intelligent and complete interpretation of the data.  Reasons for
apparent pecularities of the data that would be totally baffling
                                112

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to one unfamiliar with details of the situation and study may be
quite obvious to one who was involved in the work.  The person who
has been there can bring far more conviction to writing of the
report than one to whom the waterway is merely a picture on a map.


When final reports are prepared, they should be filed with the
agency authorizing the investigation which will, at its discretion,
make them available to other agencies and groups as deemed to be in
the public interest.


The rules and techniques of good technical writing apply the same
as for all survey reports.


     1.  Write in a clear, concise manner.


     2.  The who, what, why, when, where and how questions must be
         answered at every opportunity.


     3.  Non-committal language should be avoided.


     ESSENTIAL ELEMENTS OF A FISH KILL INVESTIGATOR'S REPORT


     1.  An Introduction that briefly tells where the kill occurred,
         when and by whom it was reported, who made the investigation
         and when, and the authority for the investigation.


     2.  A Summary description of the findings brought out by the
         investigation, including extent of the kill, and the
         numbers and species of fish involved.


     3.  Conclusions that detail the cause of the fish kill.


     A.  Recommendations that specify how similar kills may be
         prevented in the future.  Recommendations should be
         included only with sanction of, and reviewed by,
         administrators of conservation and/or pollution control
         agencies.


     5.  A Description of Study Area section that includes an area
         location map showing the extent of the kill, principal
         population centers, major waste sources and sampling
         stations.


     6.  A Fish Mortality Discussion section that details in a
         logical manner the steps taken in the investigation
         and the knowledge gained from each.
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     7.  A Water Uses section detailing the uses and value of the
         affected waterway.

     8.  A Damages section that assigns a monetary value, when
         practical, to the fish kill.

     9.  An Appendix that includes all tabular data; describes the
         manipulations used to estimate the numbers of fish killed
         and how their value (damage claim) was determined; and
         that further describes any nonstandard biological and
         chemical analytical procedures used.   If the investigation
         is prolonged and many contacts are made and investigative
         steps taken, it may be helpful to include a chronological
         schedule of all activities of the investigator pertaining
         to the fish kill from the initial report of the kill until
         the writing of the Investigation Report.  This should
         include all contacts made (when, where, how, who) as
         well as other steps in the investigation process.

Sometimes it may be necessary to prepare an addendum to the original
report if additional information is uncovered that alters conclusions
drawn or recommendations made.

                            SUMMATION

     1.  We must recognize that the report is an important and
         essential element of any fish kill investigation.  Take
         as many pains with the report as with other phases of
         the investigation.

     2.  Follow-up steps to be taken (litigation proceedings)
         may be completely based on the content of the  report.

     3.  A thorough and well-written report may save the
         investigator many headaches especially if litigation
         proceedings do occur and the investigator is called
         upon to testify.

     4.  Even if no court action is taken the report may prove to
         be useful in the event of subsequent fish kills or water
         quality investigations in the same area.
                                114

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          INVESTIGATION OF PETROLEUM CAUSED FISH KILLS

                               by

                        Sterling L. Burks

                    Reservoir Research Center
         Oklahoma State University, Stillwater, Oklahoma
This presentation on petroleum caused fish kills covers a broad
spectrum of chemical substances most of which have not yet been
isolated and identified.  Crude oil generally is not considered
to be directly toxic to fish unless it contains a high percentage
of aromatic compounds (Shelton, 1971).  Indirectly, crude oil may
smother aquatic vegetation and non-motile, benthic organisms with
subsequent indirect deleterious effects on fish through the food
chain.  Refined products such as diesel fuel, gasoline, etc. are
much more toxic than crude oil.  Many of the first surfactants or
oil dispersants used on oil spills were more toxic to aquatic
organisms than the oil; however, new products are now available
which are not acutely toxic to aquatic organisms.  Infrared
spectrophotometry (Kawahara and Ballinger, 1970, Kawahara, 1969),
atomic absorption spectrophotometry (Moore, Miller, and Glass,
1966) and spectrophotometry combined with gas chromatography
(Stuart and Branch, 1970) may be useful in identifying sources
of oil spills.

The contaminants most commonly associated with the petroleum
industry are oil, ammonia and phenols.  The standard analytical
methods for ammonia and phenol are well known and widely used.
The toxic threshold values for these substances are often a
subject of debate, especially in the case of ammonia since its
toxicity to fish is closely related to other environmental
conditions such as pH, dissolved oxygen levels, hardness, etc.

Other toxic substances are present in petroleum wastes but they
have not been adeuqately characterized nor determined to be
present in all crude oils or waste products from crude oil, an
inadequacy which is apparent in many aspects of fish toxicology.
The field of human toxicology has been established for many years
and autopsy techniques have been developed for isolation and
characterization of toxic drugs and poisons in humans, but the
field of fish toxicology is relatively new.  The fish toxicologist's
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problem is made more difficult by the tremendous diversity of
natural and synthetic chemical compounds in the aqueous environment.
A fish may absorb many compounds from the surrounding media, not
all of which are toxic.

It is a difficult task to isolate and characterize the toxic substances
from fish, and perhaps of utmost importance, determine the toxic
threshold concentration of the substance in fish.  For example,
isolation and detection of .1 ug/g (parts per million) of endrin
in the blood of a catfish would not be positive proof that endrin
caused the death of the fish although this level has been considered
to be toxic (Mount, 1966).  Analyses of acute levels of toxins in
fish is a research area of vital national importance.  Japanese
scientists have actively investigated autopsy techniques and perhaps
will develop new methods for fish toxicologists (Kariya, 1968).

We have performed many bioassays with Fathead Minnow (Pimephales
promelas) on oil refinery process waste waters and final effluents
in a cooperative research program sponsored by Oklahoma Oil Refiner's
Waste Control Council and USDI, Office of Water Resources Research
to identify ichthyotoxins.

We have analyzed test waters and fish tissues for organic toxins by
gas chromatography - mass spectrometry and for inorganic toxins by
atomic absorption spectrophotometry.  We have observed large
variation in heavy metal content of minnows purchased from commercial
dealers as compared to minnows spawned and raised in our laboratories.
Over 400 minnows have been analyzed for heavy metal content.  A
typical sample of these analyses is shown in Table I, illustrating
the individual variation.

The lack of background data on heavy metal content in native fish
creates difficulties when interpreting data from fish suspected to
have been killed by a heavy metal.  Many investigators have not
been aware of the individual variability of fish and have misinterpreted
heavy metal content of fish.  Mount (1964) utilized a ratio of zinc
content in soft tissue to skeletal tissue as an index to zinc caused
fish kills.  Such a ratio might reduce variability among fish and
permit an objective interpretation of data.
                                116

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                        LITERATURE CITED
Kariya, Teiji.  1968.  Studies in the post-mortem identification
     of the pollutant in fish killed by water pollution.  VII.
     On acute poisoning with phenol.  Bull, of the Japanese Soc.
     of Sci. Fisheries, 34:767-768.


Kawahara, Fred K.  1969.  Identification and differentiation of
     heavy residual oil and asphalt pollutants in surface waters
     by comparative ratios of infrared absorbances.  Environ.
     Sci. & Technol., 3:150-153.


Kawahara, Fred K,,  and Dwight G. Ballinger.  1970.  Characterization
     of oil slicks on surface waters.  Ind. Eng. Chem. Prod. Res.
     Develop., 9:553-558.


Moore, E. J., 0. I. Miller, and J. R. Glass.  1966.  Application of
     atomic absorption spectroscopy to trace analyses of petroleum.
     Microchemical Journal, 10:148-157.


Mount, D. I.  1964.  An autopsy technique for zinc-caused fish
     mortality.  Trans. Amer. Fish. Soc., 93:174-183.


Mount, D. I. and G. J. Putnicki.  1966.  Summary report of the
     1963 Mississippi fish kill.  Trans, of 31st North American
     Wildlife and Natural Resources Conference, March 14-16, 1966.


Shelton, R. G. J.  1971.  Effects of oil and oil dispersents on the
     marine environment.  Proc. Roy. Soc. London, B, 177:411-422.


Stuart, R. A. and R. D. Branch.  1970.  Identification of pollution
     deposits originating from petroleum oils.  Instrument News,
     21(2) 1-9.  Perkin Elmer Corp., Newsletter.
                                117

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                               Table I


Comparison of Heavy Metal Content in Commercial and Laboratory-Raised
                Fathead Minnows (Pimephales promelas)
Element


Cd
Cu
Cr
Ni
Pb
Fe
Zn
Number
Analyzed

20
20
20
20
20
20
20
Commercial Minnow
x"
mg/kg
0.83
9.80
16.15
6.39
9.46
127.20
286.66
Std. Dev.

0.56
3.90
13.42
5.03
6.26
42.73
142.26
Number
Analyzed

20
20
20
20
20
20
20
Lab-Raised Minnow
X
mg/kg
0.74
13.32
3.27
12.43
4.48
73.21
129.27
Std. Dev.

0.33
3.74
1.97
4.77
1.61
13.49
19.64
                                  118

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      TOXICANT CAUSED FISH KILL IN ALABAMA - A CASE HISTORY

                               by

                         Sam L. Spencer

           Chief of Fisheries, Game and Fish Division
    Alabama Department of Conservation and Natural Resources
During the period 1953 to June, 1968, sixteen fish kills were
reported and investigated on the Tombigbee River near Mclntosh,
Alabama.  This area is sixty river miles upstream from Mobile in
southwest Alabama.  These fish kills ranged from only ten fish
found up to 8,000.  In each case dead fish were found in the
immediate vicinity of discharges from two industries, Olin Mathieson
and Geigy Chemical Companies.  We were finally able to determine
the reason for the lower number of fish found in some kills.  One
of the industries had employees picking up and disposing of the
dead fish to help cut down on complaints from fishermen.

On June 19, 1968, a fish kill involving 50,000 fish valued at
$10,299.00 was investigated over a sixteen-mile stretch of the
river.  The kill involved six species of fish; however, bluegill
comprised 78% of the number of dead fish found.  The bluegills
exhibited reverse pectoral fins and an organo-phosphorus insecticide
was suspected.  Geigy Chemical Company manufactured Diazinon, an
organo-phosphorus insecticide.  Water samples collected after the
kill by the Alabama Water Improvement Commission (AWIC) showed
progressively higher concentrations of Diazinon downstream.  However
no sample was found to contain levels of Diazinon considered toxic
to fish.

One factor that complicated the kill was extreme water level
manipulation by the U. S. Corps of Engineers at a dam sixty miles
upstream.  The ten-year, seven-day low flow for this area of the
river has been reported to be 1,100 CFS.  The flow was reduced on
June 17-18 to approximately 725 CFS.  It was documented that all
other industries except Geigy were notified by the Corps that the
river flow was being reduced.

It was generally agreed by our Alabama Water Improvement Commission
that this kill was caused by the low water and no attempt was
made to recover damages.
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However, nine days later at approximately 9:00 a. m. on June 28,
1968, two of our conservation officers observed dead fish on the
Tombigbee River below Mclntosh and notified our Central office.
Because 1 had worked that area previously, I immediately flew in
our Game and Fish Division airplane from Montgomery to investigate
the kill.  The airplane was used to determine the exact location of
dead fish and to observe discharges of all other industries on the
river near the kill,  A visual observation of Geigy Chemical Company's
discharge was made at tree top level before landing to conduct the
ground investigations.

Two members of the Technical Staff of the AWIC were visiting the
plant at 10:00 a. m. in relation to the previous kill.  When they
arrived at the plant, they were greeted with the news of the new
kill.  We jointly investigated the kill while it was in progress.
With two boats we collected water samples and samples of distressed
fish throughout the kill area.

The water samples were iced down and fish samples were frozen with
dry ice.  Later that afternoon, after alerting the lab, the samples
were flown to the EPA Lab at Athens, Georgia.

All bluegill exhibited reverse pectoral fins and Diazinon was again
suspected.  At that time the dead fish were estimated at 1-4,000.
However, they were still dying.  The following day's investigation
by the district fishery biologists revealed 51,246 dead fish valued
at $5,857.00.

Water samples revealed Diazinon concentrations up to 200 parts per
billion.  Diazinon was found in Geigy's ditch also.  All samples
taken from the kill area exceeded the 48-hour TLM of 30 PPB and the
Federal Laboratory concluded "There was a sufficient concentration
of Diazinon in the area of the fish kill to kill fish."

Samples of control fish were collected several miles upstream from
the kill area.  Cholinesterase inhibition studies were conducted
on both batches of fish by EPA.  The average of specific activities
of both sets of samples were 0.48 (distressed fish) and 1.55 (control
fish).   This gave an average inhibition of 69%.  The EPA Lab reported
that "Dr. Charles Weiss of the School of Public Health at the
University of North Carolina reported that inhibition levels of
50.5% caused death in sunfish when exposed to organo-phosphorus
compounds in water.  Therefore, it is our belief that the cause
of death of the fish in the Tombigbee River was inhibition of
cholinesterase activity."  After considerable opposition by certain
                               120

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members of the AWIC, a suit for compensatory and punitive damages
was filed for $250,000 against Geigy Chemical Company by the State
of Alabama.

Since Geigy was the largest single employer in Washington County,
the suit was filed in Mobile County.  This was possible because
the investigators found dead fish in Mobile, Baldwin, Clarke and
Washington Counties and our law allows suits to be placed in any
county where damage occurs.

The trial was held on August 17, 1970, more than two years after
the kill occurred.  Each side took pre-trial depositions of all
persons known to have knowledge of the fish kill.  My deposition
involved two hours of questions by the opposing lawyer and resulted
in 94 type written pages of testimony.

Pre-trial testimony obtained from Geigy personnel revealed that a
plant employee had apparently opened the wrong valve and Diazinon
was allowed to enter the cooling water ditch.  This was the first
time the employee had handled this particular operation which
consisted of draining water out of a 15,000 gallon wash vessel
containing a Benzene-Diazinon mixture.  The Diazinon drained onto
the ground, travelled through a retaining dike through an open
pipe onto a gravel bed.  It travelled fifty feet through this
gravel and entered the cooling water ditch that by-passed the
treatment facility and went into the Tombigbee River where the
kill occurred.

After the law suit was filed, Geigy performed several activities
to help prevent similar occurrances in the future.

     1.  They put operating instructions on the wash vessels for
         the operators to follow.  None were there prior to the
         kill.

     2.  They closed off the open pipe through the retaining dike.

     3.  They changed the pipe and drainage system so all spills
         would be routed through the treatment basin.

     4.  Constructed piping system so cooling water could be
         immediately diverted into a five day retention basin
         if necessary.

     5.  Installed a underwater diffuser pipe and thus prevented us
         from seeing what goes into the river.
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In the interim period between the kill and the trial two reports
were received and checked out.

     1.  Geigy attempted to burn 500,000 quart aerosol insecticide
         containers just across the state line in Mississippi.
         This was true.

     2.  It was reported by several persons that seven tank cars
         of Diazinon were returned to Geigy just prior to the two
         fish kills and were emptied into the treatment basin
         resulting in an overload.  No evidence could be found
         in the railroad shipping records to support this rumor.

The fish kill and suit resulted in considerable publicity unfavorable
to Geigy.  Immediately prior to the trial Geigy offered a $25,000
settlement which we declined.  We felt we had a good case and might
possibly end up with a $250,000 award.

We employed Dr. Charles Weiss as an expert witness to testify on
the effects of Diazinon upon bluegill.   This was during the time
of considerable concern over the leaking containers of nerve gas
being dumped into the ocean.  Dr. Weiss testified that Diazinon
acted upon bluegill in a fashion similar to nerve gas upon humans.

Our district fishery biologist was questioned very closely on how
he arrived at this estimate of the number of dead fish.  Fortunately
he still had his complete set of field notes which contained the
actual counts of fish in each 100 yard segment and the location of
each sample point.  He referred to his notes and quickly convinced
the opposing lawyers that his count was probably conservative.

At the close of testimony around noon on the second day the trial
took a strange course.  The Judge stated that in his opinion, the
State had proven negligence and was entitled to the value of the
fish; however, he felt the State had failed to prove willful or
wanton destruction of the fish and, therefore, was not entitled to
punitive damages.  He stated this was probably the most interesting
case he had ever presided over; thanked the jury for listening to
the trial and directed that Geigy pay the State for the value of
the fish plus 6% interest since the date of the kill.

The award amounted to approximately $6,000.  We figured that this
probably paid for the cost of investigation, witness fees and
legal fees.  This brings up the question "Was it worth it?"  We
feel it was.  In the first place, we would have investigated the
                               122

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kill anyway.  No fish kills have been reported below Geigy during
the three years since the suit was filed.  We are now able to
collect largemouth bass in our fish samples immediately below
their discharge.


Earlier this year Geigy announced the planned construction of
additional treatment facilities that  will cost ten million
dollars to install and one million annually to operate.  There
is no doubt that the adverse publicity of the fish kill suit had
a lot to do with this industry deciding to construct this new
treatment facility.  We feel it was worth our time and effort.
We also feel this industry will be one of our "good neighbors".
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     AN ENDRIN-CAUSED FISH KILL IN ARKANSAS - A CASE HISTORY

                               by

                           Neil Woomer

         Arkansas Dept. of Pollution Control and Ecology
Arkansas is primarily an agricultural state.  The very fertile
Delta and Grand Prairie regions support "big" agriculture, and all
of the alteration and degradation of the natural environment that
is implied in that phrase is found in these regions.  Wholesale
clearing of bottomland forests, drainage of wetlands, channelization
of streams, and contamination of aquatic ecosystems by the unrestric-
ted and largely indescriminate use of pesticides and herbicides -
all of these activities have been carried on to the extent that
no stream or lake in the region has escaped some deleterious effects.

In the area of acute fish mortalities in Arkansas, those resulting
from the use or misuse of pesticides by far outnumber those from
any other cause.  An estimate of the number of fish killed by
pesticides is not possible because many of these fish kills go
unreported.  Many of the people who live in these areas simply
expect this sort of thing to happen and pay little attention
when it does.  Lately, however, an increasing number of people
are learning that they should become alarmed when this happens.
We are beginning to get better reporting of fish kills in agri-
cultural areas.  Efforts are also being made to instigate greater
controls over the use and application of pesticides in Arkansas.

In the past two or three years we have investigated a number of
pesticide-caused fish kills, some of which were more extensive
and perhaps more spectacular than the one I'm going to talk about
today.  As any of you know who have been involved in pesticide-caused
fish kills, they are notoriously difficult to pin down, and we
managed to wrap this one up quite nicely.

The fish kill occurred in Plum Bayou in mid-July 1971.  Plum Bayou
is a typical dredged stream of some 60 miles length having its
headwaters in Pulaski and Lonoke Counties in central Arkansas.
The stream meanders back and forth between these counties south
through Jefferson County to its confluence with the Arkansas River
north of the City of Pine Bluff.  It was a typical fish kill in
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that it was reported on Saturday.  A Game and Fish Biologist
contacted me at home about 10:00 A. M. with the report that a
fish kill was occurring on Plum Bayou.  The report to him was
from an individual who was concerned that his livestock which were
pastured along the Bayou would be harmed by the water that was
killing the fish.  There were rather vague references to sick and
dead cows and horses which on later investigation proved to have
no substance.

We went by the Dept. of Pollution Control laboratories in Little
Rock to get the equipment we needed for the investigation.  By
about 2:00 P. M. we were on Plum Bayou at a point about a mile
below what was later discovered to be the source of the fish
kill.  The original complainant had not left his name so we had
no contact with anyone who knew the area.

We noted the dead fish in the area, which were fairly numerous and
had been dead about a day.  We noted the condition of the water,
took dissolved oxygen, temperature, pH and chemical grab samples.
Everything looked good - in fact, too good for this type of stream,
which is typically muddy and sluggish.  The water was clear and
rather swift running with thick growths of attached algae attesting
to the fertility of the rice irrigation water that was later found
to be contributing to the strong flow of clear water.

We headed upstream to the next access point where we found the
likely source.  At this point a pump was at the water's edge with
a plastic hose leading up the bank to a clear spot along the road
where a number of discarded pesticide containers were strewn about.
A gulley was found leading over the bank to the stream which
contained a puddle of a milky yellowish substance.  This material
was sampled.  The whole area was permeated with the strong odor of
insecticides, and the ground all around was saturated with spilled
material.  The empty containers were examined.  There were a number
of gallon containers of Monosodium methane arsonate (MSMA), a
herbicide; some 5-gallon containers of Dinitro, also a herbicide;
some 5-gallon containers of about 50% Methyl Parathion; and several
5-gallon containers of an aptly named poison called Kill-Plenty,
which is a mixture of Methyl parathion and endrin.

The stream was checked upstream of this point and no dead fish were
found.  Live fish were seen.  Just below this point for perhaps 1/4
mile no dead fish were found; also no live ones were seen.  The
stream was flowing rapidly.  The flow is straight for almost a mile
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from this point.  It is dredged so that no limbs or rocks or other
objects are present to catch floating fish.  We assume that dead
fish floated downstream or perhaps the live ones fled ahead of a
heavy slug of toxicant some distance before succumbing.

We talked to some people in the area including an investigator
from the Jefferson County Sheriff's Dept. who had been called by
the people who feared that their stock were endangered.  We also
talked to two men who had been fishing in the Bayou the previous
day below the suspected source point and had seen the water become
cloudy and had seen fish in convulsions.  They had traced the
cloudy water back to the point where a farmer was filling the tanks
of his spray rig with the pesticide and using the Bayou water for
dilution.  Spillage occurred in this operation and also back-siphoning
occurred through the hose and into the stream.  We got the name and
address of the farmer who was leasing the land.  There were also
several children in bare feet running around.  They were informed
that playing in puddles of methyl parathion was likely to be quite
dangerous.  The dermal toxicity of organophosphates to humans, as
you probably know, is quite high.

The next task was to try to determine the extent of the kill and if
possible, to head off the slug of insecticide and find dying fish.
At a point two miles below the spill hundreds of dead fish were
found piled up behind a bridge pier.  All of the representative
species were noted and all sizes.  Some live fish were seen at
this point - very small sunfish which couldn't be captured.  They
were very skittish.   You could see them lying just under the
surface, with rapid opercular movements, but when you moved quickly
they darted off, zig-zagging away.  Very nervous, very scary, but
they didn't try to hide, just darted away.  The next access point
was 6 3/4 miles below the spill.  Here we hit pay dirt.  Large
carp and buffalo were found swimming at the surface, convulsive,
very distressed.  They would gulp at the surface, some would swim
in circles on their sides, some would swim rapidly to the shore
and lay at the edge, quivering and jerking.  Several of these large
fish were collected.  Blood samples were taken by the prescribed
method of cutting the fish behind the dorsal fin and collecting
blood from the dorsal aorta in clean screw-top vials, which were
immediately plunged in ice.

A water sample was taken for pesticide analysis and of course the
other standard samples.  The next access point was 8 miles below
the spill (1 1/4 miles below the last sample point).  Very few
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dead fish were found, only very small ones and more of the small,
skittish fish.  The water was muddy at this point and if one
smacked the water surface, several small fish would jump out of
the water.  The fact that small fish were dying first was, of
course, further evidence of a toxicant caused kill.

It appeared that the kill was bracketed.  We then contacted the
owner of the spray rig who was instructed to cease the mixing
operation and to clean up the spilled pesticide and empty barrels.
We had collected and tagged one of the Kill Plenty containers for
evidence.  The label clearly stated that the material was highly
toxic to fish and aquatic life and was not to be used near lakes
or streams.

Game and Fish personnel went back Monday and checked to be certain
that, the kill had stopped and found that it had extended to about
12 miles below the point of the spill.  No more fish were found
dying.   They made counts and estimated the value of the dead fish,
finding that approximately 15,000 fish were killed which were
valued at $2 ,05 i.

Analvois of the samples showed t-ndrin to be present in the water
6 3/4 miles below the spill to the extent of 5.82 ppb and methyl
parathion 8.00 ppb.   The water sample obtained 8 miles below the
spiJl contained 0.7 ppb endrin arid 0.77 ppb methyl parathion.  The
literature showed threshold toxicity of endrin to be in the range
of 0.1-0.2 ppb and TLM to be in the 0.5-1.0 ppb range depending
on species.  Methyl parathion is toxic in the range of 8 ppm.  The
blood collected from the dying fish contained 531 ppb endrin and
54.7 ppb methyl parathion.

Very little information on blood levels of fish acutely intoxicated
with pesticides is available.   The study done by the FWPCA on the
Lower Mississippi  fish kill of 1966 gives 230-280 ppb endrin as the
level in blood indicative of acute intoxication in channel catfish,
and about half that for gizzard shad.  So it appeared that endrin
was the cause of the fish I ill.  The report was prepared, and
turned over to our attorney who filed charges against the farmer.
The farmer chose not to contest our conclusions and he was fined
$250 by the judge  with $200 suspended.  No attempt was made to
recover the  value  of the fish killed.

This fish kill received very wide local coverage in the press and
even though the penalty was inconsequential I feel quite sure that

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many farmers were much more careful with their pesticide application
activities in Arkansas last summer than would  otherwise have been.
Perhaps other fish kills as serious or moresc  were prevented.  If
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  POLLUTION CAUSED BY STRIP MINING IN MISSOURI - A CASE HISTORY

                               by

                        Everett H. Fuchs

               Missouri Department of Conservation
Fish kills which cause the most problems and frustrations are not
those caused by some unknown factors, but are those which we expect.
I would like to consider situations where we are able to predict
fish kills but are powerless to prevent them; situations where an
industry announces its intentions not to cause problems, but
nevertheless, does; and situations where the cause of a fish kill
is well known and easily proven, but prosecution and collection
of damages is still difficult.

This case history starts in August, 1961, on Rocky Fork Creek,
3 miles north of Columbia, Missouri, near the geographical center
of the state.  At that time, a water quality survey of the creek
was conducted by the Missouri Department of Conservation and the
Missouri Water Pollution Board, along with representatives of
Peabody Coal Company.   The reason for the survey was the announced
intention of the coal company to start a strip mining operation in
the headwaters of the creek.

In August, 1961, Rocky Fork Creek was an unpolluted Ozark border
stream with a high aesthetic value.  It alternated between shallow
riffles and deep pools, and contained clear water and a clean
gravel bottom.  Rocky Fork contained a diverse fish and benthic
fauna.  Some of the major components of the fish fauna were largemouth
bass, green sunfjsh, bluegills, channel catfish, white suckers,
golden redhorse, and northern redhorse, along with assorted minnows
and darters.  Rocky Fork could be classed as a "wading creek".
In Missouri, a "wading Creek" is the type that is fished with light
spining gear or a light fly rod, and in an easy afternoon you
expect to catch 15 - 20 pan-size sunfish and 1 or 2 keeper-size
bass.  Rocky Fork was heavily fished and constituted a valuable
natural resource in a densely populated area.

The Department of Conservation, being familiar with strip mining
operations, feared the destruction of Rocky Fork Creek.  However,
representatives of the coal company calmed a somewhat concerned
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segment of the local population in 1961 by claiming, even in  the
newspapers, that they planned to mine without causing any damages.
There was no reason to doubt they had anything but the best of
intentions.  The mine was even named after one of Missouri's
most famous citizens, Mark Tx^ain.

Despite the best of intentions, economic considerations apparently
won out over environmental ones as the mid 1960's were marked by
poor mining practices at Mark Twain Mine.  This resulted in acid
producing spoil banks and the accumulation of acid mine water in
abandoned strip pits.  The acid water in the pits had pH values
near 3.0 and acidities near 1,000 ppm.  Also, lack of reclamation
work resulted in severe erosion from the unvegetated spoil banks.

On August 23, 1970, 9 years after the initial investigation,  a fish
kill occurred in Rocky Fork Creek.  The cause-acid mine water
flushed from several strip pits by a heavy local rain.

An estimated 17,000 fish were killed in 4.5 miles of the creek.
Their monetary value, based upon the publication "Monetary Values
of Fish", was $1,028.61.  The degradation of Rocky Fork resulting
from erosion off Mark Twain Mine was obvious during the fish kill
investigation.   Two to six inches of clay and silt covered the
gravel  bottom of many pools.

After the fish kill, the coal company brought in heavy equipment
and took some corrective action to prevent further loss of acid
mine v.ater from their property.  Needless to say, the fish were
alreadv dead.

In September 1970, Peabody Coal Company appeared in Magistrate
Court  and i1 he;;.led guilty to the charge of violating the Wildlife
Code of Missouri, Sectior 252.?IQ, which reads, "it shall be
unlawful for any person to cause any deleterious substances to
be placed, run or drained into any of the waters of this state
in quantities sufficient to injure, stupefy or kill fish 	"
Causing a fish kill is a criminal offense in Missouri and is
classified as a misdemeanor under Missouri Law.  The fine was
$500 plus costs, something less than 3 cents per fish.  The $500
fine was the maximum allowed under the law.

Seven months later, in April, 1971, a break in the dam of a coal
slurry pond blanketed 4 miles of Rocky Fork Creek with coal particles.
Officials of the coal company claimed this was an accident.  Yet,
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it was known by mine  personnel  that  the  dam was leaking and they
had even installed  a  pump  to  relieve  pressure on the dam.   After
the dam broke and the  coal  slurry  had flowed into Rocky Fork Creek,
large amounts of heavy  equipment were used to reconstruct  the dam.

During the latter part  of  1970  the strip mining operation  was
extended into the nearby Hinkson Creek watershed.   This fact was
first realized by most  Columbia citizens when the coal company
filed an application with  Columbia city  officials to strip mine
about 70 acres of land  in  the recently annexed northeast corner
of Columbia.  (' -ming  after  the  problems  on Rocky Fork Creek, and
in the new era of environmental  awareness, this request to strip
mine within the city  limits was  viewed with alarm by the local
citizens.  After a  number  of  meetings between city and company
officials, permission  to strip  nine within the city limits was
final Iv grantedj but  only  after  elaborate reclamation plans were
agreea to bv the coal  company.

On Juiv 31, 1971, a fish kill occurred in Hiriks^n Creek.  The cause-
acid -.nine water.  Approximately  300,000  gallons were drained from a
strip pit  into Ihnkson  Creek wlu n  an  employee of the coal  company
intentionally cur. the dap.  The  pit water had a pH of 2.3, an
acidity of 4,420 ppm and an iron concentration of 270 ppm.  About
one mile below the  pit  outflow  Hinkson Creek had a pH of 2.6, an
acidity of 1,900 ppm and an iron concentration of 120 ppm.  Above
the pit  outflow Hinkson (.'reek had  a pil of 7.4, an alkalinity of
115 ppr>, and an iron concentration of 0.2 -ipm.  An estimated
12,bOO fish wet(-  killed in  J.S  miles  of  the creek.   Their  monetary
value was Si,6°1.•->(,'.  Heavy deposits  of  clay and silt were present
in al ]  the pools and  the smaller riffles in the kill zone.  On
August  fi,  1971, --barges v^re  t i \-a again- t \\ .ibody Coal Company.
The cast' has n. f. vet coine  fr  trial.

On August 26, ."''I, the Missourj Department of Conservation received
a lettei from the Attornev  General's  Off ire of Missouri asking for
an escirate of damages  to  Kockv  Fork  (reek and Hinkson Creek by
pollution from the  Mark Ivain Mint-.   Indci  the Missouri water
pollution  law polluters   iia';>K  tor damages to the state caused
by wati ? p"]lulion.

The .js;,(-ssr!ieni of damages  Caused by pollution is a task in which we
had little pricr experience.  Various approaches are possible.  First,
there i,-> the monetary  value of  the pollution-killed fish.   One
method would be to  charge  lor the value  o!  the dead fish.   But what

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about the other organisms which were killed such as crayfish,
aquatic insects, and other invertebrates?  What is their value?
Do they have no value?  What about the loss of the use of the
stream itself?  Does the stream have a value?  What about the
environmental degradation, the acid water, the iron precipitates
that blanketed the bottom?

Also, long term population studies dn Missouri, on Courtois Creek,
have shown that the annual production of fish can often equal  or
surpass the instantaneous standing crop.  For example, we expect
the a:ending crop of smallmouth bass in & stretch of stream in
Septemner, 1969, would have beer  10 pounds per acre.  In September,
1970, «e would still expect the standing crop to be 10 pounds per
acre.  However, we also expect the harvest from September, 1969
to September, 1970 to equal or exceed LO pounds per acre.  It  them
becomes clear, in the fish kills under consideration, that in
addition to losing the fish that were directly killed, there was
alsi.  a loss of fish production for at least a year.  This is true
for a number of reasons.

     ;.  Not oni\  fish, but fish food organisms such as crayfish,
         aquatic insects, and even algae and diatoms were killed.
         This hinders repopulation even by fish movement from
         other sections of the stream.

     '.  Consider the time of year in which the kills occurred,
         the last day of Julv and the third week in August.
         Reproduce ion of fish was essentially completed for that
         year and there was little chance of a successful spawn
         and est I'olishment of a strong year class.

         Also, .-onsider the break in the coal slurry pond which
         oceurrt-d in April,  This not only wiped out benthic
         organisms hut prevented reproduction of fishes for at
         lease another year.

Based on the above factors the -nonetary dam iges to Rocky Fork Creek
and Hlnkson Creek were calculated.

Pollution <_f Rocky For'; Creek from Peabody Mark Twain Mine:

     .'-•igu^r 1970   Acid pollution.
                   Monetary value of lish killed         $1082.61
                   !. o
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     April 1971   Coal dust slurry pollution.

                  Prevention of repopulation of fish for

                  one additional year                     1082.61


Pollution of Hinkson Creek from Peabody Mark Twain Mine:


     July 1971    Monetary value of fish killed           1691.90

                  Loss of fish production for one year    1691.90

                                       Total Damages     $6631.63


I would like to consider, is $6,631.63 really compensation for the
damages that were caused?  I think not.  The amounts calculated by

the above method will be only a fraction of the actual damage

because it is determined for only one segment of the stream ecosystem.

Rather than trying to collect actual damages, we really need to

assess a penalty which will be an incentive to prevent damage.

Possibly the best way of doing this has already been adopted by the

State of New York.  The legal value of fish killed by pollution

has been set by the legislature as $10 per fish regardless of size

or species.   The damage assessment on the two fish kills in Rocky

Fork Creek and Hinkson Creek, based on the New York method would

be $296,000.  This would provide a strong incentive to prevent

future problems, which is the only real solution.
                               133

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REFERENCES
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              "Monetary Values of Fish", 1970, Pollution Committee, Southern
«            Division, American Fisheries Society.




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