United States
Environmental Protection
Agency
Environmental Research
Laboratory
Corvallis OR 97330
EPA-600/3-79-071
June 1979
Research and Development
&EPA
Effects of Selected
Herbicides on
Smolting of
Coho Salmon
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7. Interagency Energy-Environment Research and Development
8. "Special" Reports
9. Miscellaneous Reports
This report has been assigned to the ECOLOGICAL RESEARCH series. This series
describes research on the effects of pollution on humans, plant and animal spe-
cies, and materials. Problems are assessed for their long- and short-term influ-
ences. Investigations include formation, transport, and pathway studies to deter-
mine the fate of pollutants and their effects. This work provides the technical basis
for setting standards to minimize undesirable changes in living organisms in the
aquatic, terrestrial, and atmospheric environments.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/3-79-071
June 1979
EFFECTS OF SELECTED HERBICIDES ON SMOLTING OF COHO SALMON
by
Harold W. Lorz
Susan W. Glenn
Ronald H. Williams
Clair M. Kunkel
Research and Development Section
Oregon Department of Fish and Wildlife
and
Logan A. Morris
Bobby R. Loper
Pacific Northwest Forest and Range Experiment Station
Corvallis, Oregon 97331
Grant #R-80'»283
Project Officer
Gary A. Chapman
Western Fish Toxicology Station
Corvallis Environmental Research Laboratory
Corvallis, Oregon 97330
CORVALLIS ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U. S. ENVIRONMENTAL PROTECTION AGENCY
CORVALLIS, OREGON 97330
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DISCLAIMER
This report has been reviewed by the Corvallis Environmental Research
Laboratory, U. S. Environmental Protection Agency, and approved for publi-
cation. Approval does not signify that the contents necessarily reflect
the views and policies of the Environmental Protection Agency, nor does
mention of trade names of commercial products constitute endorsement or
recommendation for use.
U
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FOREWORD
Effective regulatory and enforcement actions by the Environmental
Protection Agency would be virtually impossible without sound scientific
data on pollutants and their impact on environmental stability and human
health. Responsibility for building this data base has been assigned to
EPA's Office of Research and Development and its 15 major field installa-
tions, one of which is in the Corvallis Environmental Research Laboratory
(CERL).
The primary mission of the Corvallis Laboratory is research on the
effects of environmental pollutants on terrestrial, freshwater, and marine
ecosystems; the behavior, effects and control of pollutants in lake systems;
and the development of predictive models on the movement of pollutants in
the biosphere.
This report describes a potentially adverse effect of pollutants on
fish such as salmon which must migrate from freshwater to seawater, and
demonstrates that under certain conditions exposure to sublethal levels of
pollutants can result in mortality when fish subsequently enter seawater.
Laboratory test methods are described which should detect this effect in
screening tests and the data obtained in this report should advance
knowledge on the effects of pollutants in aquatic ecosystems.
J. C. McCarty
Acting Director, CERL
!» •
i i
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ABSTRACT
n, I « T SeVeral herbic±de* ^ yearling coho salmon,
Oncorhynchus kisutch, were determined. All 96-h tests were conducted
under static conditions at 10°C in freshwater of alkalinity and hardness
ranging from 70-83 mg/L and 85-93 mg/L (as CaC03) . respectively. The
herbicides acroleln and dinoseb were the most toxic of the 12 water
soluble herbicides tested, having 96-h LC50 values of 68 Ld 100 ug/L,
respectively Atrazine, diquat and picloram were moderately toxic in
freshwater with 96-h LC50 values ranging from 10-30 mg/L
exhibited Z°SHd to.ABltr°lc-^ di1"at and paraquat in freshwater all
exhibited dose-dependent effects in subsequent seawater entry tests The
^^
werechall^r061 ttle °r "' *»— »t r w
were challenged with seawater. No apparent affects on the (Na K)-
a"1Vlty °£ the 8111S ~» «*««- "ith^o^f
The effect of sublethal concentrations of Tordon 101, dinoseb and
" -
January 5, 1977 to June 30, 1978
a
Deport covers the period
IV
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CONTENTS
Page
DISCLAIMER ii
FOREWORD i ii
ABSTRACT i v
FI CURES vi i
TABLES vi i i
APPEND IX TABLES x
ACKNOWLEDGMENTS xi
I CONCLUSIONS 1
II RECOMMENDATIONS 2
III INTRODUCTION 3
IV METHODS 6
SELECTION OF TOXICANTS AND EXPERIMENTS 6
EXPERIMENTAL FISH 6
EXPOSURE TO TOXICANTS 7
Toxicants 7
Static exposure tests 7
Flow-through tests 7
WATER QUALITY 10
CHEMICAL ANALYSIS 11
GILL ATPASE ACTIVITY 11
TOLERANCE TO SEAWATER 11
ASSESSMENT OF COEFFICIENT OF CONDITION 13
HISTOLOGICAL EXAMINATION 13
DOWNSTREAM MIGRATION 13
V RESULTS AND DISCUSSION 15
ACROLEIN 15
Review of Literature 15
Experimental Results 17
AMITROLE-T 19
Review of Literature 19
Experimental Results 20
ATRAZINE 22
Review of Literature 22
Experimental Results 2k
DICAMBA 25
Review of Literature 25
Experimental Results 25
KRENITE 26
Review of Literature 26
Exper i menta1 Res u 11s 27
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Page
PARAQUAT
Rev
Experimental Results
Review of Literature ..... 27
......
Review of Literature ....................... 3 1
Experimental Results ......... ..... •}•>
2 L q^T ' ~ -- ...........
Z>H»5 ' ......................................... ^l,
Review of Literature ................... [ * ' " 31,
Experimental Results ...... ......... -jr
ESTERON BRUSH KILLER ............... '.'.'.'.'.'.'.'.'.'.'." 36
Review of Literature .......... !!!!!!!!"!!! 36
Experimental Results ..... ........... -JQ
TORDON 22 K (PICLORAM) . . . ....... \\\ [ '.'.'.'.'.'.'.'.'. '.'.'. 4o
Review of Literature ............. .......'... l\Q
Experimental Results ....... ......... i.7
TORDON 101 ............... ........ ['" ........... ?^
Review of Literature ....... '.'.'. ............. 45
Experimental Results ........... ! !!'.!!!!!!!! ^6
Static exposure .................... '* ^
Flow-through exposure ...... "" L.R
DINOSEB (PREMERGE) ....... . ............ .'.'.'.'.'.'.'." H
Review of Literature ............ ........ ^9
Experimental Results ............ '.'.'.'.'. ...... 51
Static exposure ..................... ]" ci
Flow-through exposure ...... '" ct
DIQUAT ................... . ......... ^ ........... 53
Review of Literature ........ .......'.. ...... $L
Experimental Results ....... >..............' 53
Static exposure ........ "** CQ
V! GENERAL D
VII REFERENCES
VIII ADDITIONAL REFERENCES NOT'ciTED ......................
IX APPENDICIES ...... . ..... ...................... 78
APPENDIX I. METHuDOLOGY'FOR'ANALYsis'oF ........
SPECIFIC HERBICIDES ____ Qi
APPENDIX II. EFFECT OF VARIOUS HERBI ciDEs'oi''' "
HISTOLOGY OF YEARLING COHO SALMON
BY DR. J. D. HENDRICKS, OREGON
STATE UNIVERSITY ..... qo
APPENDIX III. METR.C AND ENGLISH EO,uivALENTS: '. \ '. 9°
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FIGURES
Number Page
1 Diagram of flow-through diluter ............................ 9
2 Exposure tanks with diluter in background .................. 10
3 Percent survival of yearling coho salmon during exposure to
acrolein in freshwater and subsequent survival upon transfer
to seawater ................................................ 19
^4 Percent survival of yearling coho salmon during exposure
to Amitrole-T in' freshwater and subsequent survival upon
transfer to seawater ....................................... 22
5 Percent survival of yearling coho salmon during exposure
to paraquat-CL in freshwater and subsequent survival upon
transfer to seawater ....................................... 30
6 Percent survival of yearling coho salmon during exposure
to Tordon 22K (picloram) in freshwater and subsequent
survival upon transfer to seawater ......................... M
7 Percent survival of yearling coho salmon during exposure
to Tordon 101 (2,4-D + picloram) in freshwater and
subsequent survival upon transfer to seawater .............. l\~J
8 Percent downstream migration of yearling coho salmon
fol lowing exposure to Tordon 101 .................. ......... 50
9 Percent survival of yearling coho salmon during
exposure to dinoseb (Dow Premerge) in freshwater and
subsequent survival upon transfer to seawater .............. 52
10 Percent downstream migration of yearling coho salmon
following exposure to dinoseb (Dow Premerge) ............... . 55
11 Percent survival of yearling coho salmon during exposure
to diquat in freshwater and subsequent survival upon
transfer to seawater ...................................... 59
12 Percent downstream migration of yearling coho salmon
following exposure to diquat .............................. . 61
VI I
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TABLES
Number „
Page
1 HERBICIDES TESTED FOR ACUTE TOX1CITY TO YEARLING COHO
SALMON 8
2 CHEMICAL CHARACTERISTICS OF TEST WATER DURING STATIC
TOXICANT EXPOSURE, ]2
3 ACUTE TOXICITY OF ACROLEIN TO VARIOUS FISH SPECIES 16
A SURVIVAL AND GILL ATPASE ACTIVITY OF YEARLING COHO SALMON
EXPOSED TO ACROLEIN IN FRESHWATER AND THE SUBSEQUENT
SURVIVAL FOLLOWING TRANSFER TO SEAWATER (MAY 10-27, 1977).. 18
5 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO AMITROLE-T IN FRESHWATER AND THE SUBSEQUENT SURVIv"
FOLLOWING TRANSFER TO SEAWATER (JAN. 17-FEB 16 1977) 21
6 TRAZ.N.ppu F YEARUNG COH° SALMON
TO ATRAZINE IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (MAR. 2-19 1977) '
7
8
Tn*n!!!AL AN° G'LL ATPASE OF YEARLING COHO SALMON EXPOSED
»
34
11 SURVIVAL AND GILL ATPASF OF vcam mr rni,n CAI U_M
.ING COHO SALMON EXPOSED
35
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Number
12 SURVIVAL OF YEARLING COHO SALMON EXPOSED TO ESTERON
BRUSH KILLER AND SUBSEQUENT SURVIVAL FOLLOWING TRANSFER
TO SEAWATER (MAY 31 -JUNE 16, 1977) ......................... 38
13 SURVIVAL OF BIG CREEK WINTER STEELHEAD TROUT FRY EXPOSED
TO ESTERON BRUSH KILLER, FAIRPLAY LABORATORY, OSU
(May 31-June 4, 1977) ...................................... 39
14 MORTALITY DATA FOR SEVERAL FISH SPECIES EXPOSED TO
TORDON 22K FOR % HOURS .................................... 42
15 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO TORDON 22K IN FRESHWATER AND SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (DEC. 9'27, 1976) ........... 43
16 SURVIVAL OF YEARLING COHO SALMON AT THREE FISH DENSITIES
EXPOSED TO TORDON 22K OR TORDON 22K PLUS AMMONIUM
CHLORIDE IN FRESHWATER, AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (FEB. 11-28, 1977) .......... 45
17 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO TORDON 101 IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (NOV. 30-DEC. 21, 1976) ..... 46
18 SURVIVAL OF YEARLING COHO SALMON EXPOSED TO TORDON 101
IN FRESHWATER, AND THE SUBSEQUENT SURVIVAL FOLLOWING
TRANSFER TO SEAWATER (MAR. 13'29, 1977) .......... - ......... 48
19 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO DINOSEB IN FRESHWATER AND SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (FEB. 22-MAR. 10, 1977) ..... 52
20 SURVIVAL OF YEARLING COHO SALMON EXPOSED TO DINOSEB IN
FRESHWATER AND SUBSEQUENT SURVIVAL FOLLOWING TRANSFER
TO SEAWATER (APR. 19-MAY 16, 1977) ....... .................. 53
21 ACUTE TOXICITY OF DIQUAT TO VARIOUS FISH SPECIES ........... 56
22 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO DIQUAT IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (APR. ig-MAY 5, 1977) ....... 59
23 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO DIQUAT IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER ............................. 60
IX
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APPENDIX TABLES
Number
5
Pae
1 CONCENTRATIONS OF 2,4-D AND 2,/»,5-T IN STEELHEAD TROUT FRY
FOLLOWING EXPOSURE TO ESTERON BRUSH KILLER ..!... 95
2 SURVIVAL AND GILL ATPASE ACTIVITY OF YEARLING COHO SALMON
=N?TsR'?; ;----ER.AND.S-ST".---.. 96
3 EFFECT OF TORDON 101 EXPOSURE ON AVERAGE LENGTH UFITHT
AND CONDITION FACTOR u^Hbt LtlNblH, WEIGHT,
„
6 CONCENTRATION OF DINOSEB IN VARIOUS TISSIIFS nr VCAD, ,MP
- ™
100
7 C^.SS S EXP°SURE °N AVEME LE^H, WE.GHT, AND
............................................. 101
8 pEM COH°
CH*ON,C EXPOSURE
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ACKNOWLEDGMENTS
This investigation was supported in part by the U. S. Environmental
Protection Agency, Research Grant R-80A283, and was funded through the
Corvallis Environmental Research Laboratory, Corvallis, Oregon. The U.
S. Department of Agriculture, Pacific Northwest Forest and Range Experi-
ment Station, accepted responsibility for chemical analyses of the herbicides
tested and provided the necessary personnel and equipment. Many con-
tributed to this study and their assistance is gratefully acknowledged:
Dr. G. A. Chapman, Project officer, Western Fish Toxicology Station
(WFTS), EPA, provided technical assistance and guidance. Dr. N. A.
Hartmann devised the statistical test to determine if downstream movement
of the treatment groups was statistically different from the control.
Dr. M. Newton, Oregon State University School of Forestry, provided one
of the chemicals, Krenite, used in the study. Ms. Carroll Burkett,
Forest Science Laboratory, assisted in the chemical analyses.
The authors gratefully acknowledge Drs. G. A. Chapman, A. V. Nebeker
and H. H. Wagner who provided constructive criticism on the manuscript.
xi
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SECTION I
CONCLUSIONS
1. Acrolein and dinoseb were the most toxic of the herbicides tested
having 96-h LC50 values of 68 and 100 yg/L, respectively.
2. Atrazine, diquat and picloram were moderately toxic in freshwater
with 96-h LC50 values ranging from 10-30 mg/L.
3. Fish exposed to Amitrole-T, diquat, and paraquat in freshwater exhibited
dose-dependent effects in subsequent seawater entry tests.
**. Freshwater exposure to sublethal concentrations of diquat (5 mg/L for
1A*» h) resulted in some deaths when fish were challenged with seawater.
Diquat exposures of 0.5-3.0 mg/L for 96 h resulted in reduced downstream
migration following release of fish into a natural stream.
5. No apparent effects on the (Na,K)-stimulated ATPase activity of the
gills were observed with any of the herbicides tested.
6. The herbicide formulations tested appeared to have no effect on
smelting of yearling coho salmon except for the direct toxicity of
acrolein and dinoseb, and the effect of diquat on seawater survival
and downstream migration. Therefore, application of these formulations
at their recommended levels of use (Oregon Weed Control Handbook,
1977) should not affect smelting.
7. Misuse of chemicals has led to direct loss of fish, and this suggests
that stricter enforcement of regulations and the possible need for
training and refinement in licensing of applicators is required.
8. The data indicate that some bioconcentration from the water occurred
in the fish exposed to dinoseb and Esteron; however, it is low
compared to pesticides (e.g. DDT).
9. Some of the chemicals tested produced pronounced histopathological
effects in exposed fish; the tissue effects were similar to those
produced by toxic agents.
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SECTION I I
RECOMMENDATIONS
" and
2- t
periods. herb,c,des that pers.st in the environment for extended
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SECTION I I I
INTRODUCTION
Herbicides have become a widely used and nearly indispensable tool in
reducing the competition of brushy hardwoods in intensive silviculture of
Douglas-fir in the Pacific Northwest. As Norris (1971) pointed out,
however, it is necessary to know the behavior of these chemicals to allow
their safe use, particularly with respect to water contamination.
Herbicides can enter streams through direct application to stream
surfaces, in overland flow during periods of intense precipitation or by
leaching through the soil profile. The probability of overland flow or
leaching of an herbicide to streams depends largely on the persistence and
movement characteristics of the chemical, properties of the soil, and the
degree to which precipitation infiltrates the soil surface. Leaching is a
slow process capable of moving only small quantities of chemical short
distances. Overland flow of water (and herbicide) on forest land seldom
occurs because the infiltration characteristics of the forest floor and
soil greatly exceed rates of precipitation (Norris and Moore 1971).
Norris (1967) reported that direct application of chemicals to stream
surfaces is the principal mechanism of chemical entry to aquatic systems.
This type of contamination can be prevented or minimized through the use
of buffer strips and attention to the details of application.
The opportunities for entry of herbicides from agricultural lands to
streams are similar to those from the forest except where herbicides are
used to control aquatic or streambank vegetation. Many forest and agri-
cultural herbicides are applied in the early spring and summer months when
anadromous salmonids are normally migrating downstream. A careful evalua-
tion of the possible toxic hazards of herbicides to aquatic life is
therefore necessary to insure that use of these valuable tools does not
cause toxic impacts on aquatic organisms.
Evaluation of the probability of occurrence of toxic impacts on
aquatic organisms requires a consideration of two factors. One is the
inherent toxic properties of the chemical involved, and the second Is the
probability that the organisms will be exposed to toxic amounts or concen-
trations of this chemical. The probability of exposure is related to both
temporal and spatial relationships between aquatic organisms, treated
areas, application of herbicides and the behavior of the chemical in the
environment. Behavior of chemicals in the environment includes their
movement, persistence, and fate within the aerial, terrestrial, and aquatic
portions of the environment. Of primary importance is the mechanism of
chemical entry to streams.
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Concern regarding the protection of surface waters and aquatic life
has prompted numerous evaluations of the effects of chemicals on aquatic
invertebrates and fishes. Much of the toxicological research with aquatic
biota has, however, been limited to the development of acute toxicity
values to measure the effects of the chemicals on these organisms. More
recently, chronic exposure of fish and aquatic organisms has received
attention because numerous parameters can be evaluated as indices of toxic
?Q7?CtS i MthUI" !9I2< Eat°n 197°' Macek et al" ]976a, b, HcKIm and Benoit
1971, and Mount 1968) The "laboratory fish production index" as defined
by Mount and Stephen (1967) reflects toxic effects on reproduction, growth,
spawn, ng^behay. or egg hatchability and fry survival. A parameter which
has rece.ved l.ttle attention is the effect of chemicals on seaward
m.gration and saltwater adaptation of anadromous species.
normJl! „*"* ^ H™ • rat ' ?" °f J uven ' ] e c°ho salmon (Oncorhynchus kisutch)
normally occurs dunng the spring of their second year of life. They are
fully euryhal me several months earlier (Conte et al . 1966 Otto 1971)
transfer ofTv'V^?^.3 ^Md size of 9 cm. The 'experimental
a transient hT" I S.aV?°nids fr™ freshwater to seawater Is followed by
1con?e et al iq?r M , StUrbanCe °f plasma water-electrolyte balance
caused by the Ih^i 1 ?ndSmith 1%8)' This Osmotic disturbance is
osmorequlation (H ? 9!C -Chan9eS necessarY to adapt from freshwater
^ alt "xcreHon wl / ?tl0"' ?ater excr«tlon) to seawater osmoregulat ion
the time of no™? re^nt,on). These disturbances are minimized at
! b) ^ migratlon °r "Parr-smolt transformation" (Wagner
physloloqICaTdfac?orsr ^ 'T^ data Bowing that one of the
frou t (sTl™ alTr* ^ ^^ Wlth m|9r«ory behavior in steelhead
tHpnosTha'sH^se ^ °f (Na.K)-st.fflulated adenos
^^l^^ °f (Na.K)-st.fflulated adenosine
activity doubled dur ng ?he parr- s^U%miCr;SOmeS °f 9niS' ™S *"**"*
steelhead trout (Zaugg and McLa n iqin t^f°rm5tl°n °f coh° salmon and
ATPase activity n salmnnlH, 7 ' 972' and Zaug9 and Wa9ner '973).
reaching a maximum ? " eto^^l "T^ ^^ SeaWater eXP°SUre'
factor in maintain!™ ! iZ r J ys> and IS th°ught to be an important
al 1^67 Zauaa and M • of?'1' and ionic) balance in flsh (Epstein et
sublethai fevels of rn -^l L°rZ a^d McPherson (1977) showed that
Several workers have r^6",1^-1^ the (Na 'K) "Stimulated ATPase activity.
hyd^carbon insectlriH P° ! !" Vltr°" inhibition of ATPase by chlorinated
M (Campbell et al.
hydrocarbonTnsecH"^ ^erb' ^CUrrer 3nd persist— <* chlorinated
has resulted "n^^rev? ewi" ?l£ f f^ of ^ '" ^ """"
aquatic ecosystems (Cope 1966 Eisler 1Q7^ I- ? chem.cals on
1968, and Mullison 970). Our ear Her r ElJ1?r,J?nd WaPner ]™> JohnS°n
mixtures of cadmium or z nc with loiSI rSSearf "?di"ted that copper and
coho salmon (Lorz and McPherson lS??P i detri^ntal to smolting in
studies reported here w, ! - 9??> L°rZ et a1' ]W . Therefore in the
herbicide;Pused exten^::!:6^ iorLT^ 1" ^rml^ " »« of the
effects on smolting forestry and agriculture would have similar
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This report presents data on the effects of selected herbicides on
survival, seawater adaptability, (Na,K)-stimulated ATPase levels, and
downstream migration of yearling coho salmon following acute and chronic
herbicide exposure. The report includes a detailed description of
methods, a results and discussion section arranged on a compound by
compound basis, and a series of appendices. The results and discussion
section includes reviews of pertinent literature relevant to toxicity
characteristics, and the environmental behavior of each compound at
recommended application rates.—/
I/Application rates from Oregon Weed Control Handbook 1977 unless otherwise
noted.
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SECTION IV
METHODS
SELECTION OF TOXICANTS AND EXPERIMENTS
of FiIhe,nHVWilHienftal Mana9ement Section (EMS) of the Oregon Department
of F,sh and Wildlife prov.ded a list of approximately 30 herbicides that
rPvtewedSwfthnDrr0rntai ^T in the past few years. This list was
Station) for herbicide h (Pacif]c Northwest Forest and Range Experiment
aariculture and rr^iH^k,* u ar"6 currently used in forest management and
The Forestry Science Jihor , P°tential contaminants of aquatic ecosystems.
11 ic i u i c:> L i y ^^-icnCGLSuor^tT^rw -a^-i*-«„j j_ • . *- . ~-
and fish samoles for r! °^ tOry a9reed to undertake the analysis of water
most forest sorL an ^ ?es he»-bicides chosen for study. Although in
most rorest spray appl ication<; nii-c^i..ui c ,
decided to test wate? soluble herblc?ip 1° formu atlons are used' Wf, H
simpler design and clean-up of dot n formulat,ons because they allowed
an parlipr mpi-ai
migratory disposition hlston-,^ i ^a,K)-stimulated ATPase activity,
in yearling coho sa^n fonS ^^-Is'S" "^ ™lation °f herbicides
study was limited to the smolt Mfl h- % YS exposure were examined. Our
ment Herbicide levels fo"d to Le'a Tff!^6 '" ^.^™"J* dfel°P"
may be considerably different from IK !• f °n Year1ln9 coho salmon
or fingerling salmon. Se Which could affect alevins, fry,
EXPERIMENTAL FISH
class refers to year of spawning.
6
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eggs obtained from the Oregon Department of Fish and Wildlife (ODFW) Fall
Creek Salmon Hatchery, Alsea River, Oregon, under conditions similar to those
reported by Lorz and McPherson (1977). Steelhead fry (Salmo gairdneri) from
Big Creek Salmon Hatchery (ODFW) were used in the Esteron tests in addition
to yearling coho salmon.
EXPOSURE TO TOXICANTS
Toxicants
Twelve water-soluble and one water-emulsifiable herbicides were tested
under static exposure conditions. The herbicide formulation tested, its
manufacturer, chemical name, and summary of registered uses are listed in
Table 1.
Static exposure tests
The static toxicity tests were carried out in 0.6l-m diameter fiberglass
tanks. Water was continuously aerated and 85% of the 120 L was exchanged
once per day. The fish were generally placed in the test tanks 3 days prior
to toxicant exposure for acclimatization and recovery from handling. Toxicant
solutions were mixed in a separate container prior to introduction into the
tanks. The daily exchange of toxicants always started with the control tanks
and went to successively higher toxicant concentrations; the mixing container
was rinsed following each concentration change. Upon completion of the daily
toxicant changes, the mixing bucket was rinsed several times and then flushed
overnight with running freshwater. Where possible, toxicant concentrations
tested were selected from published LC50 data for the particular herbicide.
A minimum of seven replicated concentrations, with 10 fish per test tank,
were used for each static toxicity test.
Mow-through tests
Four herbicides (Tordon 101, dinoseb, diquat and Esteron Brush Killer)
were used in a flowing water system. This system consisted of a gravity flow
diluter (Figs. 1 and 2) capable of delivering 12 L/min to each of 10 exposure
tanks (five duplicated concentrations). A volume of 1000 L was maintained in
each of the ten 1.54-m diameter fiberglass exposure tanks, and 95% of the
water was replaced every 3.7 h. Submersible pumps were used in each tank to
provide additional current, aeration and mixing. Yearling coho salmon (210-
225/tank, except the Esteron study which utilized 50 fish/tank) were fin-
clipped and allowed to acclimate at least one week before the toxicant
exposure began. The concentrations were sublethal and based on prior static
bioassays. Water and toxicant flows in the diluter were checked at least
once daily; only occasional minor adjustments were required.
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TABLE 1. HERBICIDES TESTED FOR ACUTE TOXICITY TO YEARLING COHO SALMON-/.
Common or trade name
Chemical name
Manufacturer
Registered us
00
Acrolein
Amitrole-T(CytrolR)
Atrazine (AAtrexR)
Dicamba (Banvel11)
Dinoseb (Premerge 3)
acrolein or acrylaldehyde
3-araino-l,2,^-triazole and ammonium
thiocyanate
2-chloro-ii ethylamino-6-isopropylamino-
s-trfazine
3,6-dichloro-o-anisic acid
2-sec-buty1-^,6-dinitrophenol
Diquat (OrthoR Oiquat Dibromide) 6,7-dihydrodipyrido (f,2-a:2',1'-c)
pyrazinedi?um ion
EsteronR Brush Ki1Ter
Krennite
Paraquat-CL
2,4-D (Amine 0)
2,li,5-T ('Veedar")
Tordon" 101
Tordon 22K (Picloram)
2,4-0 propylene giycol butyl ether ester *•
2,4,5~T propylene rjlycol butyl ether ester
Ammonium ethyl carbamoylphosphonate
I,1'-dimethyl-^,V-bipyridiniurn ion
2,4-dichlorophenoxyaeetic acid
(dimethylaraine formulation)
2,k,5~tri chlorophenoxyacetic ac i d
(triethyfamine formulation)
4-amino-3,5,6-trichloropicolinic
acid + 2,4-dichlorophenoxyacetic acid
both as the triisopropanolaraine salts
't-amino-3,5.&~trichloropicol inic acid
(as potassium salt)
Shell Oil Co.-Texas^/
American Cyanamid Co.
Ciba-Geigy
Velsicol Chem. Co.
Dow Chemical^
Standard Oil
(Ortho Dlv.)
Dow Chemical--7
£. I. du Pont-/
de Nemours
Standard OH
(Ortho Div.)
Diamond Shamrock
Chem. Co.
Amchem. Prod. Inc.
uow Chemical Co.
Dow Chemical Cn.
Control of submerged aquatic weeds
in flowing water.
Non-crop uses such as right-of-way,
industrial premises and ditchbanks.
Corn, sorghum, perennial ryegrass
and winter wheat.
Barley, corn, oats, wheat and
pasture and rangeland.
Both non-crop and food-crop uses.
Food crops include: alfalfa, cereal
grains, fruits, nuts and vegetables.
Non-food use; seed crops of alfalfa,
clover and vetch plus canals, lakes
and ponds.
Pasture and rangeJand, forest and
non-crop uses.
Brush control on non-cropland areas.
Preplant or directed spray on both
non-crop and food crops such as
alfalfa, fruits, nuts and vegetables.
Range and pasture grasses, vegetables,
fruit, grains, berries and certain -
aquatic sites.
On an extended basis for use on
pasture and rangeland, forests and
non-crop uses.
Pasture and rangeland
Pasture and rangeland
— CommericaJ formulations of herbicides purchased from Wiliur Ellis Co. Portland, Oregon unless otherwise noted.
^/Oregon Weed Control Handbook, 1577.
—^One liter sample provided by Shell Oil Company, Houston, Texas.
—''DOW Chemical Company supplied 5 gal. Dow Premerge 3.
S./Oregon State Dept. of Forestry (Astoria) provided 1 gal of Esteron Brush Killer,,
—^Dr. H. Jlewton, OSC School of Forestry, provided sample.
-------
Air stone.
Water inlets with
Spray heads (well, Float (connected via microswitch to toxicant manifold}
heated or chilled water) ( / /
\
-''---N-l-i
\i PI?
Waste
water
Toxicant -
Manifold
e^?
^
LV!i
Mixing —
Boxes
Distribution pipes
to exposure, .-^-~
L
L
*.*.
KJ
i\
v-!-!-%
r--i v
M—Kl
Head Box
-< i!
^S P. J>
Air stone
*
<>.
•
•
O
•. ••
&
•
_..
*
>,"
•
%
\
\
\
\
i . ,
/
/
/
Microswitch and
valve (allows
^^ dumping toxicant
|\ \. ff water or
1 v "% electricity fai/s)
;\
I
\
t \
«
%
%
\
To
waste
water
To Control Tanks
2.6 m
1.5 m
Toxicant
Pump
Covered
Toxicant
Reservoir
Figure 1. Diagram of flow-through diluter.
-------
Figure 2. Exposure tanks with diluter in background.
flowing tox«n^inmo6?r "fT* ?' ^ (ab°Ut 5°/tank) were e*po-d to
every 1 5 h I-" fiberglass tanks. Flows (5 L/min, 95* replacement
the 1 si-, tanks were provided by a siphon from mid-depth of
i OH tanKS used tor chronic pxnnQm-o ATI
prevent loss of fish. tanks were covered to
An activated carbon filter (Markin Pi,
remove herbicides from the
"
W3S Set Uf> tO
WATER QUALITY
McPherson 1977). Alkalini
the static exposure takWere
2). In the flow-through system
past
°XY9en' pH and a™ia i
" the Iaborat°rV (Table
oxygen was always >8.5 mg/L,
10
-------
PH @7 03 ammonia <0.10 mg/L and hardness @100 mg/L as CaCOj. Water
(9 0°C in March increasing to 12.3°C in June). Water temperatures were
monUored in the static and flow-through systems with continuous recording
thermometers.
CHEMICAL ANALYSIS
The concentration of each herbicide was analyzed from composite water
i n or ...
contai ^T tth^an^PproprTate' fixative. The water samples were generally
.tair.«"J^^
50 ml of teTwater At least two weeks usually elapsed between sampling
and subsequent extraction and analysis. During this period the samples
were stored in a cool area.
The analytical method for each ^^'^/^^"^tons^varild
known for the^ deviations :__^di screpanc.e^
a, behavior of the
herbicide solution in the exposure tank.
GILL ATPASE ACTIVITY
(Na,K)-stin,u,ated ATPase activity was measured on individual fl.h by
the whole gill homogenate method (Johnson et al . '^^' ( }
terization'of this enzyme assay was conducted on coho sa mon Lor^et
1978). Enzyme activity was measured at 37 C. The release g
Phosphate was measured by the ™'hod of Ern^^/J^ftrence between rates
stimulated ATPase activity was calculated as the °'™=r of 0 5 mM
of inorganic phosphate liberated in the presence or J^^J ofMry et
ouabain. Protein was measured by a ^J^ciln static tests and 10-20
al, (1951). Sample sizes ranged from 5-6 fish in static tests ana
fish in the flow-through system.
TOLERANCE TO SEAWATER
once daily.
11
-------
TABLE 2. CHEMICAL CHARACTERISTICS OF TEST WATER DURING STATIC TOXICANT
EXPOSURE, AVERAGE VALUE WITH RANGE IN PARENTHESES.
to
Toxicant
Amitrole-T
Krennite
Dicamba
Dinoseb
Tordon R101
Tordon 22K (Picloram)
Atrazine
2,4-D
2,4,5-T
EsteronR Brush Killer
Dissolved
oxygen
(mg/L)
10.5
(9.2-11.0)
10.8
(10.7-11-0)
10.7
(10.2-10.9)
11.0
(10.7-11.2)
10.0
(9.0-10.8)
10.5
(8.6-11.2)
11.1
(11.0-11.4)
10.7
(10.4-10.8)
11.0
111. 0-11.1)
10.6
(10.2-10.8)
Ammon i a
(mg/L
NH3-N) pH£/
b/ 7.5-7.6
bj 7.5-7.6
0.32 7.7-7.8
(0.22-0.42)
0.52 7-7-7.8
(0.25-0.75)
0.67 7.4-7.6
(0.65-0.72)
0.32 b/
(0.25-0.41)
0.29 7.4-7-6
(0.22-0.42)
0.45 7.2-7-6
(0.40-0.48)
7.5
(0.5-<0.7)
b/ ' 7.4
Alkal inity
(mg/L as (
79
(75-82)
79
(76-80)
83
(82-84)
81
(77-83) .
81
(80-83)
bj
84
V
77
(76-81)
80
Hardness
;aC03)
100
(100-101)
100
(99.5-100.5)
102
(101-102)
101
102
v
101
(100-101)
105
99
100
Number
of
analyses
4
4
8
4
10
12
8
4
4
1
^/pH value taken after 2 rain of gentle stirring
5/No data collected.
-------
Salinity of the water was maintained at 30 +0.5 °/°o w'th a temperature
of 10 +_ 1°C. The seawater exposure period was a minimum of 10 days; if
mortality was still occurring, longer observation periods were used.
ASSESSMENT OF COEFFICIENT OF CONDITION
Each month, after a 24-h starvation period, about 40 fish were selected
randomly from the stock and exposure tanks, anesthetized, weighed, and
measured. Individual fish were weighed to 0.1 g and fork length was
determined to 0.1 cm. The coefficient of condition (K) was determined for
each fish in the sample using the formula K = 100 W/l.3, where U denotes
weight in grams and L denotes fork length in centimeters (Hoar 1939). Any
change in condition factor compared to controls probably reflected an
effect of the herbicide on metabolic processes because the feeding behavior
appeared to be unaffected except in those fish exposed to dinoseb.
HISTOLOGICAL EXAMINATION
Gill, liver and kidney tissue were usually excised from freshly
killed fish and preserved in Bouins solution for histologicaf examination.
Three to five fish were collected from herbicide concentrations that
caused death during the static exposure. Five chronically exposed fish
were collected from each Tordon 101, dinoseb and diquat concentration.
°r. J. D. Hendricks, Dept. of Food Science and Technology (Oregon State
University) made the histological examination of the fish. Dr. Hendrick's
analyses are in Appendix II, but will be referred to in the results and
discussion whenever histopathological damage was noted for a particular
herbicide.
DOWNSTREAM MIGRATION
The effect of Tordon 101, dinoseb and diquat on migratory disposition
was assessed by releasing marked control and herbicide-exposed yearling
coho salmon into a tributary of the North Fork Aisea River and trapping
them 6.4 km downstream. The trap was checked daily for the f.rst 10 days
following release, then every second or third day thereafter. On the day
prior to release, about 100 fish from each 1.54-m exposure tank (long
exposure) and 50 fish from each 0.61-m tank (short exposure) were anes-
thetized in MS-222, weighed and marked by freeze branding and a fin clip.
No fish smaller than 11 cm were released as the "parr-smolt transformation"
is markedly size dependent. Releases were made between April 13 and May
26, 1977 the normal time of seaward migration of wild juveniles. Trapping
was terminated July 6, 1977, more than a month after the last release.
13
-------
Probability estimates for downstream migration occurring from each
release were tested for significance between the control group and each
treatment group. Significance (P=0.05) was determined by the z test where
the null hypothesis tested was:
H0: PT = PC
HA: PT * PC
PC - PT
z =
PC (I-PC) pc •
nc nT
and where: P = a/n
a = number of migrants
n = number of fish released.
A 7"km section of the stream was electrofished from above the release
sites to the weir in an attempt to collect non-migrants. No coho that had
been released earlier were found, although native cutthroat and rainbow of
comparable size to the released fish were caught. There were only two
small areas in the stream where our electrofishing gear appeared inadequate
because of the depth of the water. Thus, fish that had not migrated by
early July apparently died as a result of predators, stress, natural
causes or latent effects of the toxicant exposure.
-------
SECTION V
RESULTS AND DISCUSSION
results of tests with each herbicide are d.scussed below.
ironment
Acrole.n .. the co^n na.e for acryjaldehyde or
manufactured under the trade name °[ A<^' " ' "„ di?ches) for the
in flowing water only • —— r • - __
tshawytscha) and rainbow trout io«^ » similar to 24-h LC50 values of 75
static conditions. These findings ^.^ ^ ^ pg/L acroleln for
reported by Burdick et al. 0964 us.ng
Folmar (1976) reported the 96-n LC^O
15
-------
TABLE 3. ACUTE TOXICITY OF ACROLEIN TO VARIOUS FISH SPECIES.
Species
Ami a calva (bowfin)
Carassius a u rat us (goldfish)
Carasslus a u rat us
Fundulus similis (killlfish)
Gambusia af finis
(mosquitof Ish)
Lepomis macrochirus
Lepomls macrochirus
Lepomis macrochirus
(bluegills)
Micropterus sal mo ides
(largemouth bass)
Oncorhynchus tshawytscha
(chinook salmon)
0. klsutch (coho salmon)
Pimephales promelas
(fathead minnow)
Pimephales promelas
Rasbora heteromorpha
(harlequin fish)
Sal mo gai rdneri
(rainbow trout)
Sal mo trutta fario
(brown trout)
Toxic i ty
test
static, lab
field pond
static, lab
field tests
flowing, lab
static, lab
flowing, lab
field pond
static, lab
static, lab
static, tab
static, lab
static, lab
flowing, lab
flowing, lab
static, lab
flowing, lab
static, lab
flowing, lab
LC 5QL'
(ug/L)
62 (24 h)
2000(24 h)
1000(3 h)!/
2000(24 n)2/
240 (48 h)l/
149 (24 h)
61 (48 h)
77 (24 h)i/
2000(24 h)
140 (24 h)
100 (96 h)
183 (24 h)
160 (96 h)
80 (24 h)
1350(2 h)2/
68 (96 h)
150 (24 h)
115 Ct8 h)
84 (144 h)
140 (24 h)
60 (48 h)
5000(24)NEl/
I4o(24)i/
65 (24)
46 (24)
pH Temperature
7-2-7.3 69-72°F
69-7'ioF
7.1-7.3 69-72°F
60°F
7.2-7.3 69-72°F
7.2-7-3 69-72°F
7.4-7.7 20°C
7.4-7.6 IO°C
7.2-7.3 69-72°F
6.6-6.8 25°C
20°C
7.5-8.2 55°F
7.4-7-7 20°C
60°F
Alkalinity Hardness
89-93 '10-41
89-93 40-41
89-93 40-41
89-93 40-41
41-71
79-82 100-101
89-93 1*0-41
30 32
20
41-71
Reference
Louder and McCoy (1962)
Jordan et al. (1962)
St. Araant et al. (1964)
Butler (1965)
Louder and McCoy (1962)
Burdick et al . (1964)
Jordan et al. (1962)
Louder and McCoy (1962)
Louder and McCoy (1962)
Bond et al . (i960)
Lorz et al . (This study)
Louder and McCoy (1962)
Macek et al. (1976a)
Alabaster (1969)
Applegate et al. (1957)
Folmar (1976)
Bond et al. (i960)
Burdick et al. (1964)
-I/Concentration of acrolein causing S<)% mortality of exposed fish In time given (except as noted).
2/Total mortality in time given.
T-yTest carried out in flowing seawater.
i'40* mortality-
2/No toxic effect of concentration tested; (no information as to percent active ingredient or why compound reacted so differently).
^/Unpublished data fish pesticide lab. Columbia Mo.
-------
Ffsh exhibited a narrow range of susceptibility to acrolein. Louder
and McCoy 0962) noted the 24-h and 96-h LC50 values for largemouth bass
(Aficropterus salwoides) , bluegills, bowfins (Amia. calva) , mosquito fish
(Gambusia. affinis) and fathead minnows (Pimephales promelas RafinesqueJ
ranged from 62 to 183 ug/L. Burdick et al. (1964) found reduced toxicity
corresponding to increasing size for bluegills but not for brown trout
fingerlings.
Macek et al. (I976a) investigated the effect of chronic exposure to
acrolein on fathead minnows. These authors noted that adult fathead
minnows exposed to acrolein concentrations of 0 to 41.7 yg/L for 30 to 60
days showed similar rates of spawning, growth, and survival. However,
they observed only 2% survival of larval fish exposed to 41.7 ug/L acrolein.
The 6-day incipient LC50 for fathead minnows exposed to acrolein was 84
yg/L, and the estimated maximum acceptable toxicant concentration (MATC)
was >11.4 <4l.7 U9/L.
Under field conditions, Green (I960) found that acrolein applied at
1.0 to 2.0 mg/L could kill carp, Cyprinus carpio, and threadfin shad,
Dorosoma petenense. Largemouth bass and bluegill appeared not to be
harmed by 5-0 mg/L Aqualin, but Moen (1961) found Aqualin to be toxic to
fish at 0.5 mg/L when applied to a pond. Meyer (1961) found Aqualin was
toxic to fish when applied to ponds at rates as low as 0.2 mg/L. Catfish,
ictalurus sp. and sunfish, Lepomis sp. succumbed at 1 to 2 mg/L Aqualin,
while buffalo fish, ictiobus sp. were killed at 0.2 mg/L. Chemical control
of filamentous green algae was investigated by Jordan et al. (1962), but
they found acrolein to be erratic in its performance and the only herbicide
tested that injured fish. Furthermore, they noted 50% mortality of
bluegill and goldfish in 24 h following the third and fourth applications
of acrolein at 2 mg/L. Applications of 4 mg/L produced SB% mortality of
the bluegill and 95& mortality of the goldfish within 24 h. Placement of
fish in ponds 2 weeks after application of 4 mg/L resulted in death of all
of the goldfish and only 3% survival of the bluegills. These findings are
contrary to those of Moen (1963), but probable differences in water quality
and an eightfold greater Aqualin concentration may account for the extended
toxic condition.
During the 1960's there was interest in acrolein as a fish repellent.
Louder and McCoy 0962) reported that acrolein had been used successfully
by the iowa State Conservation Commission to drive fish downstream into a
weir in the Raccoon River. They concluded, however, that the herbicide
was not suitable for collecting fishes in lotic waters since ft required a
lethal dose to repel fish. Testing of overt avoidance reaction of rainbow
trout fry to acrolein revealed that the fry probably would not avoid a
lethal concentration (Folmar 1976).
j[xpe r ? men ta1 Res u 11 s
The 96-h LC50 for yearling coho salmon was estimated at 68 yg/L
acrolein (Fig. 3, Table 4). It was the most toxic water soluble herbicide
tested. There was no apparent effect of acrolein on (Na,K)-stimulated
17
-------
ATPase activity of the gills and little effect on seawater tolerance
following toxicant exposure (Table 4). Our 96~h LC50 value is similar to
values given by Bond et al. (i960) and Burdick et al. (1964) for salmonids.
Histological examination of fish tissues (gill, kidney and liver) indicated
that acrolein had detrimental effects which appeared to be concentration
dependent (Appendix II).
TABLE k. SURVIVAL AND GILL ATPASE ACTIVITY OF YEARLING
COHO SALMON EXPOSED TO ACROLEIN IN FRESHWATER
AND THE SUBSEQUENT SURVIVAL FOLLOWING TRANSFER
TO SEAWATER (MAY 10-27, 1977).
Concentration^/
(yg/L)
nominal
Control
5
10
20
30
50
75
100
Percent
survival—
(144-h exposure FW)
100
100
100
100
100
95
0
0
Gill
ATPase^/
M
NTH./
4.7
NT
NT
5.2
NT
NT
Percent
survival
(280-h SW)
93.8
100.0
100.0
100.0
100.0
86,7
NTSV
NT-X
2SActual concentration not measured.
— Twenty fish exposed per concentration.
—'Na ,K-activated ATPase activity of the gill; ymoles ATP hydrolyzed/mg
protein/ht mean of 4 fish.
measured, no survivors in the higher concentrations.
The toxicity of acrolein was recently demonstrated on the Rogue River
above Grants Pass, Oregon, where Magnicide H, a gaseous form of acrolein,
was introduced into an irrigation canal for the control of algae and
submerged vegetation (Oregon Department of Fish and Wildlife, notes in
Environmental Management Section file August 1977). The release of the
treated irrigation water within 2k h of treatment rather than after the
recommended holding time of 6 days (manufacturer recommendation on toxicant
container) appeared to be the cause of total mortality of fish in a 10-mile
section of the Rogue River below the spill. The Oregon Department of Fish
and Wildlife estimated that 238,000 fish were killed, including 42,000
salmonids, with an estimated value of $284,000.
18
-------
0-30,ug/L (5-30jjg/L)
100
90
_| 80
gj
> 70
^
tr eo
ID
<*> 50
2 40
LJ
0 30
(T
UJ 20
Q_
10
0
— m — (a — Qfl — EJ— BJ— 0
A 1 50 ug/L
\ \
\ \
\ \
— \ \
\ \
\ \
\ \
- k\
i 1
i I
• 1
'.\75^g/L
• 1
1 1
I!
100 \\
_ .ug/Lj L
i ^^y
— m — m — rg — BJ — 5) — (Q — -CD — 09 — D — D — CD — -Q
> \ n
B Control
K^i i i/i /I
^J\J ,UU* L
,,,..11 i 1 1 l_J
24 72 120 24 96 168 240 28O
Freshwater Seawater
EXPOSURE TIME (hours)
Figure 3. Percent survival of yearling coho salmon during exposure to
acrolein in freshwater and subsequent survival upon transfer to seawater.
AMITROLE-T
Review of Literature: Toxicity to Fish and Behavior in the Environment
Amitrole (3-amino-s-triazole) with an equal molar amount of ammonium
thiocyanate is formulated under the trade name Amitrole-T. It is registered
for non-crop land clearing for right-of-ways, industrial premises, lawn
renovation, hardwood nurseries, and ditch banks. Amitrole-T has also been
found to be very effective in the control of aquatic vegetation such as
water-hyacinths. For spot treatment, Amitrole-T is normally applied at^
1/2 Ib active ingredient (a.i.) per 12 gal of water. As an area spray it
is applied at rates up to k Ib per acre.
The literature contains many publications describing the fate of
applied amitrole. In a study of 55 different California soils, Day et al.
(1961) reported that amitrole disappeared rapidly within 2 weeks^after
application. Rapid decomposition of amitrole was also observed in a
similar study in Oregon soils CFreed and Furtick 1961). Sund (1956) noted
that amitrole adsorbs rapidly and tightly to soil particles having a high
base exchange capacity and a high organic matter content. Amitrole complexed
readily with metals in soil.
19
-------
Riepma (1962) found that soil microorganisms decomposed amitrole
rapidly and that the decomposition rate increased with increasing amounts
of soil organic matter. Grzenda et al. (1966) observed a tight adsorption
of amitrole to the hydrosol and a fairly rapid decomposition of amitrole in
a pond. Norris (1970) found that the common brush control herbicides,
2,4-D, amitrole, 2,4,5-T, and picloram were all degraded in the forest
floor although the rates of degradation varied considerably. In red alder
(Alnus rubra) forest floor material, 80% of amitrole was degraded in 35
days.
Amitrole's persistence in water, following aerial application to
forest areas for brush control in Oregon, has been studied in several
investigations (Marston et al. 1968, Norris 1967, Norris et al. 1966, and
Tarrant and Norris 196?)• In a study of a stream in a coastal, municipal
watershed sprayed with 2 Ib/acre amitrole for control of salmonberry, the
maximum concentration of herbicide, 155 ppb (in water),was observed 30 min
after application (Marston et al. 1968). Only 26 ppb was detected after 2
h and none after 6 days. Norris (1967) monitored amitrole concentration in
a stream at a spray site treated at 2 Ibs/acre. Five min after spray,
*t22 yg/L amitrole was detected; 10 hours later the concentration had
decreased to 4 yg/L and none was found after 3 days.
The toxicity of Amitrole-T is thought to be similar to that of ammonium
thiocyanate (House et al. 1967). Russian studies have indicated that
ammonium thiocyanate is lethal to fish at 200 mg/L (Demyanenko 19^*1 )•
Amitrole-T is believed to be relatively non-toxic to fish (Meyer 1966).
Two-inch bluegills tolerated 10 mg/L Amitrole-T for 100 h (U. S. Fish and
Wildlife Service 1963). Sanders (1970) noted bluegills survived concen-
trations of >100 mg/L Amitrole-T for 48 h.
The literature contains a number of publications on the toxicity of
amitrole formulated without ammonium thiocyanate. Bond et al. (i960) found
the *t8-h LC50 for coho salmon to be 325 mg/L amitrole under constant flow
conditions and water pH 7-5 to 7.7 and 1*1 to 71 mg/L total alkalinity.
Largemouth bass were able to survive 1,000 mg/L amitrole under static con-
ditions, but in flow-through apparatus all test fish died at this concen-
tration in 6 days. A 24-h LC50 of 1,200 mg/L amitrole was established for
blueglll sunfish by Hughes and Davis (I962a) in water of 29 mg/L hardness
and pH 6.9.
Hiltibran (1967) investigated the effects of selected herbicides on
fish reproduction. Survival of fertilized eggs and fry of bluegill, green
sunfish (Lepomls cyanellus), smallmouth bass (Micropterus dolomieu) and
lake chub sucker (Erimyzon sucetta) were not affected by 50 mg/L amitrole
under static conditions.
Experimental Results
The 96-h LC50 of Arnitrole-T was approximately 70 mg/L for yearling
coho salmon (Table 5). There was no apparent effect of the herbicide, on
the (Na.K)-stimulated ATPase activity of the gills, however, upon transfer
to seawater there appeared to be a dose-dependent mortality effect (Fig. A,
Table 5). Our 96-h LC50 value is one-third of that reported in the Russian
20
-------
studies for ammonium thiocyanate (Demyanenko 1941). The reason for the
discrepancy is unknown although differences in water quality or species
tested are known to be important variables. Histological examination of
dying fish showed degenerative changes occurring in the liver, kidney and
gill of fish exposed to 200 mg/L (Appendix II). The deaths observed in
seawater generally occurred within the first 24-48 h following transfer
from the toxicant and probably were not directly related to impaired
osmoregulatory ability.
TABLE 5 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO AMITROLE-T IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (JAN. 17"FEB. 6, 1977).
Concentrat
(mg/L)
nominal
Control 0
0.25
0.50
1.0 0
25.0
50.0
100.0
200.0
ion
measured
.03-0.06
0.20
0.37
.89-0.97
24.8
48.6
104.5
200.7
Percent
survival—/
(144-h exposure FW)
100
100
100
100
100
55
14.6
0
Gill
ATPase*/
2.1
2.1
NT£/
2.7
2.9,,
2.4^
NT
NT
Percent
survival
(336 h SW)
100
100
100
100
56.3
12.5
0
NT
— Twenty to 21 fish exposed per concentration.
-/ (Na,K)-stimulated ATPase activity of the gill; mean of 5 fish.
c/
— Not measured.
-'Mean of 3 fish.
21
-------
100
_| 90
> 80
IE 70
W 60
t^ 50
O 40
20
10
0
0-25mg/L
\ x:-x—
—X- X X X—
0-lmg/L
-x x x x x—x
25mg/L
_ 200mg/L
J
50 mg/l
J 1 ,1
192
240
24 72 120 24 120
Freshwater Seawater
EXPOSURE TIME (hours)
Figure 4. Percent survival of yearling coho salmon during exposure to
Amitrole-T in freshwater and subsequent survival upon transfer to seawaten
ATRAZ I NE
Review of Literature: Toxicity to Fish and Behavior in the Environment
Atrazine (2 chloro-^t ethylamino-6-isopropylamino-s-triazine) is a wide
spectrum symmetrical triazine herbicide. It is widely used to control many
broad leaf and grass weeds in the production of corn, macadamia nuts,
pineapples, perennial ryegrass, sorghum and winter wheat. Atrazine is also
employed in some areas for selective weed control in Christmas tree farming,
grass-seed production, highway right-of-way clearance, and conifer refores-
tation. Hall et al. (1972) noted that more than 100 million pounds of
atrazme is applied annually to agricultural lands in the United States.
Atrazfne Is not as strongly bound to soil particles as is the triazine
nerb.c.de^imazine. Its lower solubility and reduced adsorption add to its
mobility in the soil. Axe et al. (1969) studied the residual life of
atraz.ne in the soil and found that only 33* of the herbicide remained
after 5 days. Under most climatic and edaphic conditions, atrazine (2-4
^??re)Dha?/e?ldual Phytotox'city for k-J months (Harris and Sheets
1965). Residual carryover after repeated application is minimal.
22
-------
Analysis of surface, subsurface, and finished waters in Iowa, where
atrazine is widely used in corn production, indicated that the herbicide
was present in small amounts (<50 yg/L). Atrazine levels in surface waters
correlated with discharge volume data for the river (Richard et al. 1975).
Laboratory and field tests have indicated that atrazine is moderately
toxic to fish in comparison to other herbicides. Macek et al. (I976b) in-
vestigated the effects of atrazine on survival, growth, and reproduction of
three species of fish. Utilizing soft water (hardness 33 to 40 mg/L) and a
continuous flow apparatus, Macek et al. were able to show in acute toxicity
tests that both the 96-h and incipient LC50 for fathead minnows were
15 mg/L atrazine (95% Cl 11-20). Their acute 96-h LC50 for bluegills was
>8.0 mg/L and the incipient LC50 was 6.7 (5-4-8.4) mg/L atrazine, which
agrees with the 96-h LC50 of approximately 6 mg/L atrazine (wetable powder)
reported by Walker (1964) for this species. The 96-h LC50 for atrazine
toxicity to brook trout reported by Macek et al.. (I976b) was 6.3 mg/L (4.1-
9.7) and the incipient LC50 was 4.9 mg/L (4.0-6.0). This is similar to the
48-h LC50 (12.6 mg/L) reported for rainbow trout in a static bioassay
(FWPCA 1968).
Bluegill and fathead minnow spawning, survival, and growth were not
affected by exposure to 0.095 and 0.213 mg/L atrazine, respectively (Macek
et al. 1976b). Hiltibran (1967) found that 10 mg/L granular atrazine did
not affect green sunfish embryo development, or bluegill and green sunfish
survival over 8 days. Lake chub sucker fry (Ermyzon sucretta) survived
10 mg/L wetable powder atrazine. Similarly, brook trout parental survival,
egg production, and hatchability appeared to be unaffected by exposure to
<0.72 mg/L atrazine (Macek et al. 1976b). Survival and growth of brook
Trout fry were, however, significantly reduced following 90 days of
exposure to 0.72, 0.45 and 0.24 mg/L atrazine. Analysis of muscle tissue
from bluegills, fathead minnows, and brook trout indicated that these fish
bioconcentrated detectable amounts of atrazine after prolonged exposure
(Macek et al. 1976b).
Walker (1964) observed no fish mortality after application of 2.0 to
6.0 mg/L atrazine to ponds infested by aquatic weeds. He suggested,
however, that atrazine had the potential to affect fish in ways other than
direct toxicity. A reduction in bottom fauna was observed immediately
following application. Among the most sensitive species were mayflies
(Ephemeroptera), caddisf1ies (Tricoptera;, leeches (Hirudinea) and
gastropods (Musculium). Studies by Macek et al. (1976b) on the chronic
toxlcity of atrazine to selected aquatic invertebrates 'indicated that
morphological development of progeny is particularly sensitive. Exposure
of two successive generations of chironomids to 0.23 mg/L atrazine resulted
in reduced hatching success, larval mortality, developmental retardation,
and a reduction in the percentage of pupating larvae and emerging adults.
Continuous exposure to 0.25 mg/L atrazine significantly reduced production
of Daphnia. Development to the seventh instar of F] gammarids exposed to
0.14 mg/L atrazine was reduced 25% below that of lower concentrations and
controls.
23
-------
Herbicidal destruction of aquatic vegetation may expose small forage
fish to predation by large predacious fishes. Furthermore, fluctations in
oxygen tensions have frequently been shown to occur after application of
triazine herbicides. These fluctuations were associated with phytoplankton
blooms occurring in conjunction with decomposition of submerged vegetation
(Walker 1964).
Experimental Results
In our toxicity test, atrazine (AAtrex) appeared to produce a gradual
concentration dependent mortality in freshwater, with losses of 5 and 25%
at concentrations of 8 and 15 mg/L, respectively (Table 6). Dead fish
showed signs of severe edema. No apparent affect on (Na,K)-stimulated
ATPase activity occurred. McBride and Richards (1975) found that atrazine
significantly decreased sodium uptake of isolated perfused gills from carp
(Cyprinus carpio) but did not affect fluid flow rate at the concentrations
tested. The authors indicated the effect of atrazine on (Na,K)-ATPase
systems should be studied, because other pesticides (aldrin and DDT) have
been shown to inhibit (Na+K)-ATPase (Cutkomp et al. 1971, Koch et al.
1971). When the survivors were transferred to seawater, the group that had
been exposed to 15 rng/L atrazine suffered a 25% mortality (Table 6). The
deaths occurred within the first 24 h and probably resulted from the poor
condition of the fish (due to toxicant exposure) and not osmoregulatory
failure. Histological examination of three fish exposed to 15 mg/L atrazine
showed no apparent affect on liver or kidney tissues but hypertrophy of
gill epithelium was evident in two of the fish (Appendix II).
TABLE 6. SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON
EXPOSED TO ATRAZINE IN FRESHWATER AND THE SUBSEQUENT
SURVIVAL FOLLOWING TRANSFER TO SEAWATER (MAR. 2-19, 1977).
Concentration
(mg/L) Percent survival-/
nominal measured (144-h exposure FW)
Control
0.25
0.50
1.0
3.0
5.0
8.0
15.0
0
0.21-0.23
0.48-0.51
1.31-1.38
3.85-4.25
4.47-5.62 -
10.65
18.0-18.8
100
100
100
100
100
100
95
75
Gill
ATPase^/
2.8
NT£/
NT
3-4
NT
NT
2.4
2.3^
Percent
survival
(264-h sw)
100
100
100
100
100
100
94
75
^/Twenty fish exposed per concentration.
-' (Na,K)-stimulated ATPase activity of gills; mean of 5 fish.
— Not measured.
1/M,
Mean of 3 fish.
24
-------
DICAMBA .
Review of Literature: Toxicity to Fish and Behavior in the Environment
Dicamba (3,6-dichloro-o-anisic acid) is one of the most extensively
used benzoic acid herbicides in the United States. Production of dicamba
in 1971 was estimated to be 6 million Ibs of active ingredient (Lawless et
al. 1972). The herbicide was introduced in the early 1960's for the
control of preemergence and postemergence broadleaf weeds in cereal grains
(Frear 1976 and Velsicol Chem. Co. 1967). Phenoxy-tolerant broadleaf weeds
and brush species are also controlled by foliar and area applications of
dicamba. Current registration for this herbicide is for use on barley,
corn, oats, wheat, pasture and rangeland, where it is applied at rates
ranging from 1/4 to 8 Ib/acre. The dimethylamine salt of dicamba, formu-
lated as a liquid, is sold under the trade name of Banvel, but is also
available in granular form as the acid or amine salt.
Dicamba has been shown to exhibit intermediate persistence in many
soil types when compared to other herbicides (Burnside et al. 1971),
remaining phytotoxic for several months (Klingman and Ashton 1975). It has
a relatively high water solubility (7900 mg/L) and has been demonstrated to
move within the soil profile with water flux. Studies by Trichell et al.
(1968) have shown that runoff losses of dicamba are limited. The vapor
pressure of dicamba is quite low, so minimal amounts are lost through
volatility (Montgomery et al. 1976). Photodecomposition of dicamba is
similarly limited. Indirect evidence from a number of studies suggests
that microbial degradation may be instrumental in reducing dicamba per-
sistence (Frear 1976 and Hahan et al. 1969)- Morris and Montgomery (1975)
noted that following spraying of a brushy area in coastal Oregon with 1.12
kg dicamba/ha, water residue levels rose sharply to 37 ppb about 5 h after
spraying and then declined slowly to background levels by 37.5 h.
•
The effects of dicamba on fish have not been well investigated.
Toxicity tests with dicamba indicate a low toxicity to salmonids and warm
water fish species. In static tests conducted by Bond et al. (1965) the 2k
and 48-h LC50 values for juvenile coho salmon were 151 and 120 mg/L active
ingredient, respectively (methyl orange alkalinity approximately 55 mg/L,
pH about 7.7). Rainbow trout were killed by a concentration of 320 mg/L
dicamba in 72 h (Bond et al. 1965). Bohmont (1967), in his literature
review of Cope's 1962 and 1963 work, however, reported an estimated 48-h
LC50 for rainbow trout and bluegill of 35-0 mg/L and 130 mg/L dicamba
(Banvel D), respectively. Hughes and Davis (1962b) reported 24 and 48-h
LC50 values of 600 and 410 mg/L for the liquid formulation of dicamba for
the bluegill; however, when Banvel D acid was adsorbed onto vermiculite
they found a 24-h LC50 of 20 mg/L.
Experimental Results
No mortalities were observed in yearling coho salmon exposed to dicamba
(Banvel) concentrations up to 100 mg/L (Table 7). Gill (Na,K)-stimulated
25
-------
ATPase activity appeared unaffected by the herbicide. Histological exami-
nation of gill, liver, and kidney tissue indicated no apparent effect of
exposure to dicamba (Appendix II). Following 144-h exposure to the toxicant,
the yearling coho salmon were challenged with seawater. Fish previously
exposed to the lowest concentration, 0.25 mg/L dicamba, showed a 32%
mortality during the 11 days of the seawater challenge (Table 7), however,
no deaths occurred at higher concentrations. The deaths began after 96 h
in seawater and concluded at about 200 h. No explanation is available for
this unusual pattern of mortality. The deaths followed the time pattern of
osmoregulatory failure noted in previous seawater challenge tests following
copper exposure (Lorz and McPherson 1977). '"nuwuiy
TABLE 7. SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO DICAMBA IN FRESHWATER AND SUBSEQUENT SURVIVAL FOLLOWING
TRANSFER TO SEAWATER (JAN. 6-23, 1977).
Concentration
(mg/L)
nominal
measured
Control
0.25
0.50
1.0
5.0
10.0
50.0
100.0
0
0.19- 0.22
0.40- 0.42
0.54- 0.56
3-15- 3.33
10.05-10.11
50.5 -53-2
108.2-109.9
100
100
100
100
100
100
100
100
ITwenty fish exposed per concentration.
-(NaK)-stimulated ATPase activity of gillsf mean of 5 fish.
—Not measured.
Gill
ATPase*/.
2.52
NT£/
NT
NT
1.64
NT
2.0k
2.20
Percent
survival
(268-h SW)
100.0
68.4
100.0
100.0
100.0
100.0
100.0
100.0
KREMITE
Review of Literature: Toxicit
to Fish and Behavior in the Environment
gal water for spot treatment. 3 5 lb/acre or * lbs/100
Krenite
inactivated
has a high water solubility
in the sot, (Oregon Weed Control Ha^o^'W7)?'
is nonvolatile, and
is rapidly
Soil residue
26
-------
studies have indicated that, in 2 weeks, half of the herbicide is converted
to carbamoyl phosphonic acid which is subsequently converted to C02
humic acid fractions within 8 weeks. Bottom
months or less (Dr. James Harrod, duPont Co.,
sediments lose Krenite
unpublished report).
and
in 3
There is little published data on the toxicity of Krenite to fish.
Unpublished findings of E. I. duPont de Nemours and Company, Inc. (technical
pamphlet) indicate that rainbow trout and fathead minnow have a 96-h LC50
of 1,000 mg/L (product), while bluegill sunfish exhibit a 96-h LC50 of 670
mg/L (product). Laboratory tests have demonstrated that Krenite is not
bioaccumulated. Residues in fish tissues were comparable to the concen-
tration of the herbicide in the water (Newton and Norgren 1977).
Experimental Results
No mortalities were observed in yearling coho salmon exposed to
Krenite concentrations up to 200 mg/L. When survivors were transferred to
seawater only minimal mortality occurred (Table 8). Krenite is not very
toxic to coho salmon, as neither freshwater survival nor subsequent seawater
survival were affected. There was no apparent effect of Krenite on the
(Na.K)-stimulated ATPase activity of the gill (Table 8).
TABLE 8. SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO KRENITE IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (FEB. 2-17, 1977).
Concentration
(mg/L)
nomi nal
Control
0.25
0.50
1.0
10.0
50.0
100.0
200.0
Percent
survival—/
(144-h exposure FW)
100
100
100
100
100
100
100
100
Gill
ATPase*/
2.2
1.8.
r1/
NT-'
1.7 -
NT
NT
NT
2.7
Percent
survival
(216-h SW)
100
93
100
100
100
100
100
100
—Twenty fish exposed per concentration.
-/ (Na,K)-stimulated ATPase activity of gills; mean of 5 fish.
—'Not measured.
PARAQUAT
Review of Literature: Toxicity to Fish and Behavior in the Environment
Paraquat is the common name for 1,I'-dimethyl-4,4"-bipyridiniurn ion,
It is formulated under the trade names OrthoR Paraquat and GramozoneR.
27
-------
Paraquat formulations are presently available as chloride salts (Lawrence
et al. 1965). The pure chloride salt is a white solid that forms a dark
red aqueous solution. Paraquat is a nonselective, quick-acting herbicide
and des.ccant, and is used extensively due to its effectiveness on grasses
and most broadleaf weed species (Calderbank and Slade 1976). Paraquat is
registered for use as a directed spray on preplant treatment in many tree
crops and several f.eld crops including alfalfa, corn, soybeans, sugar
beets, and tomatoes. It is also employed as a preharvest deslccant on
cotton, potatoes, and soybeans, and in aquatic weed control. Normal
application rates for paraquat are 1/4 to 1 Ib (a.i.)/acre, and 0.1 to
i. 5 mg/L in the aquatic environment (Calderbank and Slade 1976).
U * ?Uart?rnarV ammonium salt, highly soluble in
organic solvents, and nonvolatile. The herbicide
r
ponds at rates of 2.1 and 2.5 mg/L persisted in the water for 6 and
"-
Burnet (1972) found that amphi
were
muscle. Upon transfer of the fish to fre^hwa^r ?h ^^ bUt nOt ln the
declined. Similar findings for whole-body res dues ^f T uT *?""*
were reported by Cope (1966). When severaPnnnH qU3t and Paraquat
paraquat (emulsion) or diquat oaraau^ M P°nds were treated with 1 mg/L
bluegill sunfish (1.21 mg/L whofe-bodf r bl?accur!atlon Was detected in
accumulation was detected (0^9 mg/L)" ™*^ ^ Httle di"uat b'-
trout, green sunfish, and channel catfish ,£ i para<'uat in ra!nl>ow
0.37 mg/L or less. cna™ei catfish (Ictalurus punctatus) were
Colorado pond. Residue 1 eve s
8 days after treatment and tnen decined!
Bluegtlls, largemouth bass fahead
trout exhibit approximate ihrLhotd
ter treatment of a
3 maX?lhum °f K58
levt!s w;th fish
app'cation '» a remote
in
bility.
28
-------
paraquat (cation) for a 96-h contact period (Lawrence et al. 1965). Davis
and Hughes (1963) reported a 48-h LC50 for bluegill sunfish as
100 mg/L paraquat (cation). Under static conditions (pH 7.6 to 8.0, hardness
210 to 290 mg/L), brown trout had a 48-h LC50 with paraquat of 82 mg/L
(Woodiwiss and Fretwell 197*0. Alabaster (1969) reported a 48-h LC50 for
harlequin fish as 32 mg/L paraquat using a flow-through technique and water
of 20 mg/L hardness. Butler (1965) exposed the estuarine longnose killifish
(Fundulus similes) to 1.0 mg/L paraquat and found no effect.
Information on paraquat toxicity to fish under field conditions
is limited. Yeo (1967) reported that smallmouth bass and mosquito fish
were killed when placed in ISO-gallon plastic pools with 1.0 and 3-0 mg/L
paraquat (pH 9.A). Blackburn and Weldon (1962), however, reported that
paraquat applied to a small Florida canal at 1.0 mg/L was not toxic to fish
(water temperatures 29 to 33°C, PH 7.6). Earnest (1970 noted that a
minimum of 34% of bluegills, placed in a Colorado farm pond, died within
48 h after treatment with 1.14 mg/L paraquat (surface temperature 18.9-
25.0°C, alkalinity 69-129 mg/L and pH 8.0-10.4).
Newman and Way (1966) reported no direct effects of paraquat or diquat
on aquatic invertebrates in their experiments. They did, however, note
severe oxygen depletion at one location following decay of the treated
aquatic weeds. A low oxygen content is postulated to have caused the
deaths noted among Hirudinea, Isopoda, Odonata, Coleoptera, Tnchoptera,
Gastropoda, Lamellibranchiata and captive trout, although free-living
coarse fish and trout appeared unaffected.
Experimental Results
In our study the 96-h LC50 of paraquat-CI was 76 mg/L for yearling
coho salmon. Deaths occurred in a dose-dependent manner depending on
concentration and exposure time (Fig. 5). When surv.vmg fish were
transferred to seawater, all coho salmon previously exposed to 50 mg/L died
during the first 40 h of exposure. Similarly, 64% of the fish previously
exposed to 10 mg/L died during the first 68 h in seawater (Table 9) Our
96-h LC50 value of 76 mg/L is slightly lower thar.the 48-h LC50 of 82 mg/L
given for brown trout by Woodiwiss and Fretwell (1974). The different
species and water quality, however, of the two test solutions could account
for this. The (Na.K)-stimulated ATPase activity of the gil s was not
affected by exposure to paraquat-CI. Histological exam.nation of fish
exposed to 100 mg/L paraquat-CI for 120 h showed ev.dence of degenerate
damage to gills, and kidneys, and slight necrot.c areas in the liver
(Appendix II). Earnest (1971) similarly reported necrosis'of liver tissue
following treatment of the Colorado farm pond with paraquat.
29
-------
0-lmg/L 0-lmg/L
100
90
_l 8°
^ 70
«•••
r5 60
LL.
Z>
(/> 50
2 40
LU
0 30
CC
CL 20
10
0
— x--r-3|-T-x x-y-x :
1 \ \ lOmg/L
i » ^
\ \ \
\ \ \
I ' V
i \ |00 T50mg/L
• /^^rtO/!_ J\.
\ x> *
i \ \
\ \ \
\ %» \
^ \ K
\ h
\
\200 \
_ Lmg/L v
>t
1 1 |\. 1 frx*
•y— X X X X X X X X X X
\\
\\
n
i \
i \
I q IOmg/L
\ \
' \
l \
i Y, rt
1
T
\50mg/L
\
V
** fZ 120 48 120 168 240
Freshwater Spnu/nter
EXPOSURE TIME (hours)
Figure 5. Percent survival of yearling coho salmon during exposure to
paraquat-CL ,n freshwater and subsequent survival upon transfer to seawater
TABLE 9. SURV
G'LL ATPASE OF YEARLING COHO SALMON EXPOSED
. L 'N FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (MAR. 22-APR. 7 1977).
Concentration
(mg/L)
nominal measured
Control
0.25
0.50
1.0
10.0
50.0
100.0
200.0
i ii i • i .
0
0.1*»- 0.19
0.50- 0.52
1 .08- 1.12
10.6
56.3
112.7-113-7
238.7-251.2
Percent
survival^/
exposure
100
100
100
100
95
35
0
0
-Twenty fish exposed per concentration.
°f
Percent
Gill survival
ATPaseV (240-h SW)
3.0^
NT£/
k.k6
1.96
1.90
NT
100
100
100
100
36
0
NT
NT
30
-------
Review of Literature; Toxicity to Fish and Behavior in the Environment
The chlorine-substituted phenoyxacetic acids, 2,M> (2 A-dichlorophenoxy-
acetic acid) and 2,4,5'T (2,4,5-trichlorophenoxyacetlc acid) were .introduced
as selective herbicides following investigation of their growth-regulat.ng
and herbicidal properties during World War II (Templeman 1955). They are
effective against many broadleaf weeds but not against gram.naceous weeds.
The herbicide 2,4-D is registered for use on forests, range and pasture
grasses, rye, wheat, barley, oats, corn, asparagus, apples, pears, grapes,
potatoes, blueberries, cranberries, non-cropland uses, turf, and certain
aquatic sites.
Most phenoxyalkanoic acid herbicides are formulated as the salt or
ester form. Amines of 2,^4-D are the most commonly used salt forms of
2,l»-D, although the sodium, potassium and ammonium salts are also used.
The di-and tri -substi tuted amines are particularly important; they are
highly soluble in water and are used in the formulation of water so uble
concentrates. Normal rates of application of 2fW> amines are as follows:
1A to 1-1/2 Ib/acre (a. I.) for most crops; 2 to 3 Ib/acre (a.i.) for
forest spraying; and 0.25 to 0.50* (a.i.) for the control of emergent and
floating weeds. The ester formulations of 2,4-D are extremely .mportant
and are used extensively in forestry and agriculture for vegetation control.
The ester formulations are 10 to 100 times more toxic to fish and other
aquatic organisms than the dimethylamine salt, but are often chosen for^use
because of desirable physical properties such as: control of droplet size,
limited solublity in water, good spread of herbicide upon contact with
vegetation being controlled, and persistence in the environment.
The salts of 2 k-D have very low volatility. When dissolved in hard
water they form insoluble and essentially inactive calcium, magnesium, or
iron salts of 2,4-D. Leaching of 2,M> in soil is dependent upon so.l type
and herbicide solubility in water. Decreased leaching occurs in clay and
organic soils due to adsorption by soil colloids.
Crosby and Tutass (1966) found that 2,4-D decomposed rapidly in the
presence of water and ultraviolet light or sunlight, and they identified
the decomposition products. Audus (1950) demonstrated the .mportance of
microbial degradation of phenoxyacetic acid herbicides. In a warm, moist
loam, 2,4-D can be expected to disappear within 2 to 3 weeks (Loos 1976).
Morris (1970) noted that 3k% of 2fM> was degraded in 35 days when applied
in a red alder forest floor.
Wojtalik et al . (1970 found no harmful or distinguishable response or
accumulation in zooplankton, phytoplankton or macroinvertebrates following
treatment of water with 20 to kO Ib per acre (acid equivalent, a.e.) of
DMA-2.4-D. The authors believed there was little danger of biomagnif ication
of the 2,4-D in contrast to chlorinated hydrocarbon pesticides. Wojtalik
et al. reported residues (in water) greater than 0.02 mg/L at only 2 of 19
stations J» weeks after treatment. The authors stated that disappearance of
DMA-2,/*-D from water is rapid compared with the herb.c.des dichloben.l and
fenac which persisted at detectable levels for up to 160 days.
31
-------
and fUh fnM d.methy amine salt of 2,4-D (DMA-2.4-D) in hydrosol
Harmln (iq?il? M'P aPPl|cation hav* been reported by Schultz and
wtth el ier 2 2k iVor 8 ifi I £*** Ge°rg!a' and MiSS°UrI Were treated
tectab e llvei of A n % kg/ha °f the herbicide. The highest de-
aool ication ?I f '5 '? Wat^ (°'692 mg/L) was found 3 days after
DMA-2 k-b !h.\P°K '!! u°?!a that had been treated with 8-96 kg/ha
ma/L) 28 H^v, !f^ ^ d decreased to trace amounts (less than 0.005
S^T1"1'3^^^^
seven days after treatment. Largemouth bass, channel catfish, bluegill,
and xftT/^A Ish'Lepomis microlophus, were held in live cages during
Fifteen Lr^n/TIr^-5 u° morta]]^ "** observed in any of the ponds.
2 J-D M n tn n mn /, ? fmPled contained detectable residues of DMA-
m^t»h i- 2'?!° mg/k?)' Radlometric measurements of the uptake
water hvdrJn? dlS"1P?ti°n of.the DMA salts of ring-labeled C^-2>-D in
water, hydrosol. and f,,h are in general agreement with these findings
2!S-
>attish- ••"«>'
muscle (Schultz 1971) ' le>Pylonc «eca>kidney>nver>gin>brain
residues were found for onty tort'perioSs'of ?f7> -°ted 'hat h^^"e
adjacent to forest treatment areas r I ? '" streans "ithin °r
have been found ,„ uddST' "" '' '"' m9/
ears
exceeded 0., mg/L n
A review of the toxtcity of DMA-2 4-n h^rk: -^ - ...
are relatively low in toxicity to fish ?„? n«S, indlcates that
LC50 for rainbow trout at liS'mg/L Lis IndrH( ^ t^^ 9 96"h
and Davis (1963) tested the toxiciiy of <^f?e r"^65 ( 963) and HugheS
bluegills. They found considerab e ^artaifon ?n th01?"1" °nS °f 2^"°
formulations and even in the toxicity of a sini? f* tO*lc]t* of different
researchers felt that these i neons stenctes °rmula^°n. The
different batch lots and/or
32
-------
and the dimethylamine formulations were the least toxic to bluegills of 11
formulations of 2,4-D tested (800-166 mg/L as A8-h LC50 depending on
batch). The isopropyl ester and butyl ester were the most toxic (0.8 and
1.3 mg/L as 48-h LC50 concentrations, respectively, for bluegi lls) .Davis
and Hardcastle (1959) found differences in LC50 values for 2,4-D and other
herbicides when waters from two different sources were used m toxicity
tests.
Schultz and Harman (197^) reviewed the literature concerning aquatic
use of 2,4-D compounds as a prerequisite for registration of 2,4-D for use
on irrigation canal banks and for use in moving water. They noted that
many formulations of 2,M> are available, but the .0"« "»s* JfTJ ^""SL
in aquatic situations is the dimethylamine salt of 2,4-D (DMA-2,4-D). The
ester formulations have also been used, but are many times more toxic to
fish and other aquatic organisms than the dimethylam.ne salt. Meehan
et al. (1974) tested the toxicity of various formulat.ons of 2 4-D to
salmonlds and noted that <50 mg/L 2,4-D acid produced no mortal , ty_ except
in pink salmon fry. The butyl ester, however, was very toxic <*"s'"9
almost complete mortality in all species at concentrations >1 mg/L The
isooctyl ester was the least toxic of the ester formu ations. M^han
concluded that specific phenoxy herbicide ester formulations should be
chosen with regard to their impact on aquatic organ, sms if there ,s a
possibility that the chemical will enter the water.
Rodgers and Stalling (1972) exposed rainbow trout, channel catfish
and bluegills to a C^ labeled butoxyethanol ester of 2.A-D and studied
Us u "ke from the water by fed and fasted fish Maximum res.due concen-
trations were observed in most organs of fed f.sh w.th.n 1-2 h of exposure
and w?th in -8 "exposure for fasted fish. The herbicide or its metabo-
lites were eliminated rapidly after maximum residue concentrations were
'
10 months after treatment.
Histological and biochemical changes were observed in bluegi 11 sunfish
" ! -D, Este ron 39 C
f
. --
deposits in the blood vessels and stasis and engorgement of the brain
circulatory system.
Experimental Results
, A mortalities were observed when yearling coho salmon
In our study, no mo^aj'*l^/^ 2,4-0 (DMA) for MA h (Table 10).
were challenged w.th up to 200 mg/L of z . activity of the gills
Similarly, no affect on the lNa,K st ™u d to 2 i,.D were challenged
or our test results to those of -,—...-•• -- - - desired
would be safest to use the DMA formulat.on of 2,4-0 if the desired
effects could be achieved.
33
-------
TABLE 10. SURVIVAL AND GILL ATPASE ACTIVITY OF YEARLING COHO SALMON
EXPOSED TO 2,4-D IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (DEC. 29, 1976-JAN. 14, 1977).
Concentration
(mg/L)
nominal measured
Control
0.25
0.50
1 .0
10.0
50.0
100.0
200.0
0
0.25- 0.18
0.52- 0.59
1.07
10.4
59.6- 64.0
106.8-115.4
170.0-237.0
Percent
survival—'
(144-h exposure
100
100
100
100
100
100
100
100
Gi11 „/
FW) ATPase^-7
1.4
NTH/
NT
1.6
NT
NT
1.0
1.3
Percent
survival
(240-h SW)
100.0
94.8
100.0
100.0
100.0
100.0
100.0
100.0
— Twenty fish exposed per concentration.
-'(Na,K)-stimulated ATPase activity of the gills; mean of 5 fish.
—'Not measured.
2,4,5-T
Review of Literature: Toxicity to Fish and Behavior in the Environment
The herbicide 2,4,5-T (2,4,5-trichlorophenoxyacetic acid) is identical
to 2,4-p with the exception of an additional chlorine atom on the number 5
carbon in the ring. Many of the roses, legumes, and broadleaved plants
that are res.stent to 2,4-D are controlled with 2,4,5-T. The herbicide has
been cleared for use on pastures, rangelands, right-of-way, and forests.
rt is not reg.stered for use on food crops nor for use in water, on ditch
hTrh^lHTfc ST- °r recjftional areas> °r "ear populated areas. The
herb.c.de 2,4,5-T ,s normally applied at 1 to 2 Ib/acre (a.i.) for area
treatment." ^^ ^'''^ Per 10° 9a11otlS of solution for spot
n/rf ft S°lub111ty °f,2'2|'5:T <23« mg/L) is less than that of 2,4-D
mg/L) (Montgomery et al. 1976)], and this influences its pattern of
he sodium salt of 2,4,5-T, for example, unlike that of 2,4-" has a
lubility and ,s difficult to get into a herbicidally act ve solution.
use. The
low sol
-cium,
Under condttrons favorable for microbial degradation 2 4 5-T has a
-
following application, and there was l«s ?han 0 02 in/h •"
1 year or 0.76* of the originally applted 245-7 0?K remain"s
ester and amfne formulations are'coSpa able to ttese of'z S
of the
-------
Data on toxlclty of triethylamine salts of 2,4,5-T to aquat.c organisms
are scarce. Kenaga (1974) reviewed the literature concerning the tox.city
to fish of 2,4,5-T and its derivatives. Exposure to the commercial formulation
DED-WEED* at concentrations >72 mg/L (a.e.) for 2k and 96-h resulted in at
least 50* mortality of bluegill sunfish, channel catfish, and fathead
minnows. Exposure of rainbow trout to the same formulation [>72 mg/L_DED-
WEEDR (a.e.)] resulted in 502 mortality in 2k h whereas at concentrations
of 0.07-0.72 mg/L (a.e.) for 96 h exposure Kenaga also reported mortality
rates of 50*. Spot (Leiostomus xanthurus) exposed to 0.4 mg/L (a.e.; tor
2k h showed no mortalities. The fathead minnow exposed to 40.2 mg/L for
72 h had no mortality, but at concentrations >72 mg/L for 2k or 96 h at
least 50* mortality was observed (Kenaga 1974). Day.s and Hughes (1963),
utilizing the triethylamine salt of 2,4,5-T (Crop R.der*) found the 2k and
48-h LC50 for bluegill sunfish to be approximately 53 mg/L (a.e.;.
Experimental Results
No deaths were observed in yearling coho salmon exposed Jo concen-
trations of 6-7 mg/L of 2,4,5-T (trlethylamine salt, We^ar) for 1^ h
(Table 11). Similarly, there was no apparent effect on (Na.K) -stimulated
ATPase activity of the gills. When the fish were placed in seawater only
minimal mortality occurred (Table 11). The lack of mortality durmg the
toxicant exposure was surprising considering that concentrations of 0 07-
0.72 mg/L (a.e.) were reported to cause 50* mortality m 96 h to ra.nbow
trout (Kenaga 1974); however, no water quality data were presented
Generally, LC50 values for herbicides obtained at 2k h have been about 10
times greater than the LC50 concentration noted for 96-h exposure.
TABLE 11 SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED TO
TABLE 11. SURVIVAL ANU^ ^ FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER (MAR. 31-APR. 17, 1977).
Concentration
(mg/L)
nominal measured
Control
0.05
0.10
0.25
0.50
1.0
10.0
0
0.016-0.021
0.041-0.043
0.13 -0.15
0.27
0.57 -0.74
6.64 -7-0
Percent .
survival—
(144-h exposure FW)
100
100
100
100
100
100
100
Gill
ATPase^/
4.06
NT£/
NT
NT
NT
3.52
4.66
Percent
survival
(240-h SW)
100
95
100
100
100
100
100
S/Twenty fish exposed per concentration.
K)-stimulated ATPase activity of the galls; mean of 5 fish.
measured.
35
-------
ESTERON BRUSH KILLER
Review of Literature: Toxicity to Fish and Behavior in the Environment
o ; ro Kil!er c?ntalns ^^1 concentrations of 2,4-D and
2,4,5-T [2 Ib each/gal (a.e.)] as the propyleneglycol butyl ether (PGBE)
ester. This ester of 2,A-D and 2,J»,5-T is very soluble in oil and organic
solvents but has very low solubility in water. However, in the presence of
emuls.fiers and with agitation, an emulsion is formed in water; Esteron
Brush Killer is applied in this manner.
The 2,4-D esters of low-molecular-weight alcohols have an appreciable
vapor pressure Loos 1976). The long-chain alcohols with an ester linkage
animal" V°1at'1e a"d present less of a hazard to non-target plants and
?s-orPtIon of these ester formulations is also
of nvdrolv - °f de9radat!on to the parent compound. The speed
are ^ r 3 °- ' '^ 6SterS ^ been related' to PH' Alkaline conditions
condU ons f Ken^ K7J?CSt;rlf !CatI°n of these compounds than are acidic
s?ud ed dLrS T 97 ^ I6aSley and Wmian* (1969, in Kenaga 197*)
ester forSSS !" ° ^number of 2,4,5-T esters including the PGBE
slit ar ratlf «? * ? ™t*r °f pH 6'5 a11 °f the herbicides exhibited
X s rates ?or% ['n'"^00 ^ * cont-'e^ration of I mg/L (a.e.).
l-T «tlr« nf 2'1|-J1esters should be faster than those observed for
.i) i esters o
-T «tr« nf 1
.i) i esters of comparable structure.
respectively (FWPCA 1968). 2,4,5-T was 980 and 570 yg/L,
int?
pyl
may
and chemical properties The l-T S t^*™** in thelr P^slc
cients, whichare rel«ed to the grelter abL ^ "'^ partition coeff
fatty tissues such as skin and gills The RGB? £ °t ^ comP°unds i
esters of 2,/^,5-T have relative?! hlnh > ? ' BE> butyl ' and 'sopro
partially explain th.lr^S1^c^h^J*;J7^«|§l-Jt.. which m
Sublethal effects of PGBE esters nf 9 L n u
fish (Cope 1966). Spawning of blulgin su^f7 h ^ demonstrated for
ponds treated with 5 and 10 mq/L of ih h I •JWaS delaVed 2 weel
-------
Much of the fish toxicity work on the phenoxy herbicides concerns the
PGBE esters of 2 4-D or 2,4,5-T, but little has been done on mixtures of
these compounds!' Hughes and Davis (1962b) found that both the 24 and W-h
LCSO's for bluegill sunfish exposed to the PGBE ester of 2,4-D were 29 mg/L
under static conditions in water with a mean PH of 6.9 and a mean hardness
of 29 mg/L. Meehan et al. (1974) observed that the 96-h no-effect evel
for coho salmon fingerlings exposed to the PGBE ester of 2,4-D was less
than 1 mg/L. There was a mean fry mortality of 26.7^ after 96 h of
exposure to 1 mg/L of the herbicide in water that ranged in hardness from
10.0 to 33.6 mg/L as calcium plus magnesium. A 48-h LC50 of 1.1 mg/L PGBE
ester of 2,4-D was reported by Cope (1966) for rainbow trout (no water
quality given). Butler (1965) observed that the 48-h LC50 for the
estuarine longnose killifish was 4.5 mg/L in seawater.
Studies on the toxicity to fish of the PGBE ester of 2,4,5'T have been
reported by Kenaga (1974). This herbicide appears to be generally more
toxic than the corresponding PGBE ester of 2,4-D. All rainbow trout died
within 24 h at 0.13 mg/L (a.e.) of the PGBE ester of 2,4,5-T, and all f.sh
were dead after 3 h of exposure to 0.67 mg/L of the herbicide (no water
quality given). After 7 h of exposure to 0.13 mg/L of the (PGBE) Reddon"
formulation, all bluegill sunfish were dead. Bluegills exposed to Esteron
245 (PGBE) had 24- and 48-h LCSO's of 17 mg/L in static water with a mean
PH of 6.9 and a mean hardness of 29-0 mg/L. Exposures of fathead minnows
to different formulations of the PGBE ester of 2,4,5-T [0.13 to .33 mg/L
(a.e.)] all resulted in 100* mortality within 72 h (no water quality
given) .
Bioconcentration of the PGBE ester of 2,4-D has not been observed in
fish tissues. No detectable residues of the herb.c. f were found n
bluegill sunfish exposed to 10 mg/L PGBE ester of 2,4-D (Cope 19&6).
Mat Ida et al (1975) noted that when a mixture of 2,4-D and 2,4,5-T as
the bu?ox fethanol 'ester" (commercially called "Brush Killer" "asaer a Y
spread over 9-5 hectares of forest at a rate of 150 kg/ha, no appreciable
change was noted in the aquatic community. The ?«thors were unab e to
detect the chemical in the stream during the 48 h observation period
following spraying. Similarly, fishes (cherry salmon and dace f.ngerlmgs)
showed no mortality nor abnormal behavior and the J^'J^™^1^ a]
vertebrates appeared unchanged. In a later laboratory st udy ^^a et al.
(1976) found that "Brush Killer" (mixture of 2,4-D and 2,4 5 T) exnib ted
toxic effects on aquit Ic sow bugs Asellus hilgendorffii and cherry salmon
I '
considered a nonspecific response to a toxic agent.
37
-------
Exp_g_r_i_menta 1 Resul ts
No deaths occurred In yearling coho salmon exposed to Esteron Brush
Killer for 96 h at <_800 ug/L (nominal concentration under static conditions)
nor in the flow-through exposure tanks (210 yg/L maximum nominal concen-
tration). No deaths were observed in the subsequent seawater challenge
tests (Table 12). In both the static and flow-through systems the measured
amount of Brush Killer was very low even though extra care was taken in the
mixing of the toxicant solutions. The reason for the low recovery is
unknown.
TABLE 12. SURVIVAL OF YEARLING COHO SALMON EXPOSED TO ESTERON
BRUSH KILLER AND SUBSEQUENT SURVIVAL FOLLOWING
TRANSFER TO SEAWATER (MAY 31 -JUNE 16, 1977)
Concentration
(yg/L)
nominal measured
A. Static exposures
Control
1
7
25
75
150
300
450
800
3.6-
1.9-
2.4-
7.0-
.7-
30
45.0
72.6-
104.1-
147.7-159.0
2.6
2.8
7.2
35.4
51.0
73.6
111.6
B. Flow-through exposures
Control
35
70
140
210
0
23.7
41.4- 45.7
59.6- 74.0
84.3- 88.3
Percent
survival—/
_(96-h exposure
100
100
100
100
100
100
100
100
100
100
100
100
100
100
FW)
Percent
survival
(290-h SWJ
100
100
100
100
100
100
100
100
100
(15)^
(20)
(15)
(20)
(15)
(20)
(15)
(10)
(10)
100 (20)
100 (20)
100 (20)
100 (20)
100 (20)
-
through.
or kidney tissue. The qi 1
curved gin filaments? ep tel
in some lamellae (Appindlx M)
Steel head trout fry were al
„ younger
P° apParent affect on either l
,the fo]^'^ abnormalities
hypertrophied, and aneurisms
38
-------
In the first exposure test approximately 50* of the fry exposed to 1200 yg/L
(nominal concentration) for 48 h died (Table 13 A whereas ,n a ^st 2
weeks later, only a single death was recorded at 1200 yg/L ^en though the
test was run for 96 h (Table 13 B. Unfortunately the water samples from
tSef^t So and 1200 yg/L groups were lost during "tract, on. Thus « *>
not know if total concentration of the herb.c.de were s.m, ar be™f " ™*
two tests As was noted in the test with yearl.ng coho salmon, the measured
Sncen r^ion of Brush Killer was only a fraction of the expected. Reasons
for the discrepancy in both the nominal and measured concentrates of the
herbicide and deaths observed between the two tests are unknown.
TARIF 11 SURVIVAL OF
TABLE 13- ™L°F
BIG CREEK WINTER STEELHEAD TROUT FRY
STERON BRUSH KILLER, FAIRPLAY LABORATORY,
OSU (MAY 31 -JUNE 4, 1977)-
Concentration
(yg/L)
nominal measured
A. May 31-June 4, 1977
Control
1
7
25
75
150
450
300
1200
32.9-34.0
12.1-12.3
13.2-13-5
17.7-17.9
35-2-37.2
43-9-44.3
77.4-80.1
B. June 12-16, 1977
Control
800
1000
1200
6.5- 7.2
202.0-269.0
240.0-243-0
226.0-246.0
Exposure
96 (60)
96 (60)
96 (60)
96 (60)
96 (60)
96 (60)
96 (60)
48 (60)*
48 (63)
96 (60)
96 (60)
96 (60)
96 (60)
Percent
survival
(range)
100
100
100
98.3 (100-96.7)
100
100
100
88.3 (100-76.7)
50.8 ( 80-24.2)
100
98.4 (100-96.7)
98.4 (100-96.7)
98.4 (100-96.7)
nun^er of fish exposed in replicated tanks under static conditions.
„**, -icant ensure initiated in
afternoon of same day.
Dr. D. Woodward, U. S. Fish
Research Unit (personal c^u;lcatl,r ut fry.
with the PGBE ester of 2,4-D for cutthroa trout Try
toxicity to be affected by PH or water hardness. in
Woodward and Mayer (1978) noted that surv.va } °
was significantly reduced when they were cor 1 '
be 31
He did not find
publication
a H
concen.
yg/L
-D PGBE ester'
39
-------
Following a simulated field application Woodward and Mayer (1978) recommended
that water residues of the ester should not exceed 100 yg/L after a single
application nor 31 yg/L following multiple applications.
Histological examination of the steelhead trout fry showed liver and
kidney tissues to be normal, but gill tissue (including controls) showed
evidence of hypertrophy of the chloride cells (Appendix II).
Examination of steelhead trout fry showed evidence of 2,4-D and
2,4,5-T residues following exposure to Brush Killer (Table A-l). The whole
body residues generally increased with increasing concentration and
exposure time. Fish that were moribund or dead exhibited the highest body
residues (Table A-l). Fish placed in clean water for 48 h following 48 h of
toxicant exposure showed slightly reduced residue levels.
TORDON 22 K (PICLORAM)
Review of Literature: Toxicity to Fish and Behavior in the Environment
the ™tt (J"am'no:3.5,6-trlchloroplcoUnic acid) or picloram is one of
h Plant 9r°Wth ^lators currently employed
There are three basic formulations: potassium salt
ester- ToLn 22K
0o -
are lost through volati 1 iza ion P?c oram h^K ''T am°UntS °f
extensive photodegradation b^u traviotet Uoht H T J° Under9°
solutions or soil surfaces (Hall et a 1.1 968) Sl">l'9ht in aqueous
Although degradation of nirlnram :^ i
(National Research Council Can'ada 974 NR^ '] temper?te cli-tes
does occur by nonbiological processes (Han^iqAQ?0^1''0" ln the S°H
colloids is minimal in neutral oTalLnn" ! ^ }' Ms°W*™ to soil
with decreasing PH, increasing organic conten! \nd™ SOll'.but '"creases
trations of hydrated iron and'aluminium oxides' (Foy
Cement of the
Picloram through rainfall and leach inaT'C C°n^nt- The removal of
leachmg ls one of the major factors governing
40
-------
its dissipation under field conditions (NRCC 197*). This mobility is also
of environmental concern, as leached picloram may be transported to aquatic
ecosystems such as ponds, lakes, and streams. Residue levels 'J surface
runoff have reached 2 mg/L following application at 1 . 1 kg/ha (NRCC 97*).
However, studies have indicated that under most conditions only small pro-
portions of picloram (less than 5%} applied to a watershed are transported
in surface runoff.
Only negligible residues of picloram occur In streams, apparently due
to rapid dilution of the herbicide (Haas et al. 1971). Field plots
adjacent to the mouth of a small stream were treated with 1.1 kg/ha of
Picloram, and water samples were collected 0, 0.8 and 1 .6 km downstream
from the plots following each rain for 5 ninths after application. P^°J™
was detected in the stream samples only during the f , rst significant runoff
(0.029 mg/L). No residues were found in subsequent samples (Haas et al.
1971).
Picloram contamination in lakes has not been reported, but levels In
farm ponds adjacent to plots treated with 1.1 kg/ha picloram reached 1 mg/L
(NRCC 1974). Dissipation of the herbicide In ponds has been shown to be
rapid. One study found an initial decline of U to 18* of p.cloram per
day, followed by a decline of less than 135 per day 15 weeks after appli
cation (Haas et al . 1971). Residues of picloram in the pond-bottom sediments
(H8 yq/kg) immediately following appl i cat ion were only twice that m the
water (Kenaga 1973, In NRCC 1974). After 75 days, residues of 7 ^Ag
Picloram were detected in the pond-bottom sediments and 0.1 yg/kg p.cloram
was found in the water.
It is apparent from a number of studies that the toxicity °J
to fish is influenced by its formulation and the water qua , ty (NRCC
Woodward 1976, Sergeant et al. 1970). Technical gra de P,c lora m (a .
was found to be more toxic under alkaline conditions (Woodward 1976).
Increasing the PH from 6.5 to 8.5 increased the tox.c.ty to cutthroat and
lake trou? by a factor of 2 in both species Increasing temperat ^ did,
but increasing hardness did not, lead to an increase m toxicity (Woodward
1976).
The acute toxicity of picloram varied considerably with the formu-
lation and fish species. The isoocytl ester of P'^^PP68^ J° £ *£
most toxic commercial formulation (NRCC 197*. Sergeant et al. 1970, Kenaga
1969) LC50's reoorted for this formulation are approximately mg/L for
sensU.iveCs ecieT Tc^ty levels of Tordon ^K (potassium sat) are
considerably lower as shown for several fish spec.es in Table If.
Based on available information, chronic f
not cumulative in terms of lethality (Woodward_ 976, NRU, _
exposures however have been shown to affect fish development and growth
(Woodwar 'l^Kand swimming response and liver histopa thology ( ergean t
et al. 1970). Woodward (1976) observed that the no-effect concentrat.on of
technical grade picloram for lake trout was apparently <35 P9/L, as this
level of herbicide reduced fry survival and growth. Most mortal t es
occurred during yolk absorption, which took 4-5 days longer in p.cloram
treated fish.
-------
TABLE 14. MORTALITY DATA FOR SEVERAL FISH SPECIES EXPOSED
TO TORDON 22K FOR 96 HOURS^/.
Fish species
Water temperature
Concentration
(mg/L) a.e.
Percent
mortality
Black bullhead
Bluegill
Brook trout
Brown trout
Fathead minnows
Green sunfish
Lake emerald shiner
Rainbow trout
50
65
50
50
50
50
69-73
50
S.k
91
69
52
22
29
22
91
39
30
58
22
-Modified from Kenaga (1969).
-Calculated or derived 96-h LC50 values.
-^Highest concentration producing no mortality.
50
0
50
50
0
50
0
50
0
50
0
50
50
0
formulation of picloram (for
death (Sergeant et al. 1970).
transfer to clean pond water.
shortened the recovery times"; ,,,
fish failed to recover. Analyt
sunfish swimming behavior. Ser
grade and commercial
_ or the 22% commercial
I hour) caused imnrabi1ization but not
of normal swimming response fo..-
!"bse3uent exposures to the herbicide
er a fourth exposure, many of
picloram did not affect green
oworkers suggested that technical
Picloram might contain a toxic impurity
technical picloram (1.2 mq/L) Xr ?CH °m green sunf'sh exposed to
reve^ed
.
cells (Sergeant et al. 1970)
disappearance of
tubular or smooth
panied by liver enlargement.
T reve^ed abnormalities in these
"ltrastructural changes involved the
'nCreaSe In
ChanQeS Were also accom"
en. ncreasin t
from 1 to 5 days did not alter thl , ?! 9 exposure time of the herbicide
ter the ul trastructural pattern.
-------
Residue analyses of aquatic organisms exposed tp picloram "
that this herbicide is not bioconcentrated in invertebrates or along food
chains (NRCC 1974). Daphnia exposed to 1 mg/L of the potass'um salt of
Picloram had whole body residues of the herbicides equal to that present ,n
the water (Hardy 1966). Bioconcentration of pic oram (acid) was not
evtdSt in mosquito fish exposed to 1 mg/L (a...) for IB days Youngson and
Meikle 1972). The concentration factor for these f.sh on a wet ^.ght
whole body basis was only 0.02. The 18 days of exposure to p.cloram was
adequate to achieve a steady state level of accumulate in the mosqu.to
fish.
Experimental Results
In our study, the 24-h LC50 of Tordon 22K was
- -rr__4. of picloram on
Histological
K fmu]ated
seawater after exposure to this low Tordon
seawater mortalities at low concentrations occurred
101 and Dicamba exposures, and they also were not ex pi a
tanks were used for seawater challenge tests for Jhe three chemicals
oxygen concentrations were @9 mg/L when the mortalities occurred.
FOLLOWING TRANSFER TO SEAWATER
nom i na1
Concentration
(mg/L)
Percent
survival—/
exposure
Gl11 h/
ATPase^-7
Control
0.25
0.50
0.75
1.0
5.0
15.0
30.0
0-0.12
1.67
1.89
2.76
3.89-^.23
10.54-11.84
31.81-41.23
37-95-45-10
100
100
100
100
100
100
NT
NT
2.2
1.8
Percent
survival
(288-h SW)
100
75
100
100
100
100
NT
NT
, ,*wwicy fish exposed per concentration. .
f' (Na,K)-stimulated ATPase activity;. mean of 4 fish.
•*•/ ]\T/™\ ^~ WljCI. -* «V • V H* J^ J
measured.
dead in 48 h.
dead in less than 8 h.
43
-------
100
90
< ^
> TO
—> 60
50
40
CO
LU
O
LU
Q.
20
10
0
0-5mg/L
-x—x x—x x-
(92
240
^ _
24 72 120- 48 96 ^4"
Freshwater Seawater
EXPOSURE TIME (hours)
Figure 6. Percent survival of vearlinn ,. h
Tordon 22K (picloram) in fr^h.,!* S salmon during exposure to
to seawater. rresnwater and subsequent survival upon transfer
tration as possible factors9^? t° tGSt f'Sh density and ammonium concen-
following exposure to low concentra?r«» * ^served seawater mortality
the toxicant, survivors were n?»r ! ' °f T°rdon' Following exposure to
apparent ill effects or death^ !, '!! Seawater and survival monitored. No
various fish density and ammoni^ obse^ed in seawater tests of the
therefore, did not appear ™b&]g™T^Bb]e 16) - Se^ater deaths,
levels during toxican? exposure ^ W'th fish denslty« or
-------
TABLE 16.
SURVIVAL OF YEARLING COHO SALMON AT THREE FISH DENSITIES
EXPOSED TO TORDON 22K OR TORDON 22K PLUS AMMONIUM CHLORIDE
IN FRESHWATER, AND THE SUBSEQUENT SURVIVAL FOLLOWING
TRANSFER TO SEAWATER (FEB. 11-28, 1977).
Fish
density
No. /tank
10
20
5
10
20
10
20
10
20
5
10
20
5
10
Concentration (mg/L)
Nominal
Tordon
Control
Control
Control
0.05
0.05
0.10
0.10
0.25
0.25
0.25
0.50
0.50
0.50
0.80
Measured
_
-
-
0.058
0.048
0.11
0.11
0.50
0.42
0-32
0.59
0.71
0.67
0.89
NH/jCl
(added)
0
0
0.80
0
0
0
0
0
0
0.80
0
0
0.80
0
.
3" ~
0.34
0.70
1.05
0.25
0.64
0.38
0.63
0.23
_ f _
0.65
1.01
0.22
0.61
1.04
0.27
•i— — . " •
Percent survival
144-h
exposure FW
-/Measured ammonia level, average of six daily values.
100
100
100
100
100
100
100
100
100
100
100
100
100
100
260-h SW
100
100
100
100
100
100
100
100
100
100
100
100
100
100
TORDON 101
«•»» W I ^ I l.^l«Ji.vii *•*"•' ~' i i i ---- - -••
The Dow Chemical Company's Tordon* 101 formulation contains 5-7* •-•-
(0.54 Ib/gal) tri isopropanolamine salt of P'cl^am plus 2 .2* a .e ^
(2.0 Ib/gal) 2,4-D as the *•"? Isopropanolamine saU. The
Picloram with 2,4-D enhances the translocat.on of 2 £0 £ornpet i t ; Ve .
1976). Other reported interactions are el^nr,a^e^fa° weeds, woody
Tordon 101 is used in the control of ^annua and perenmal^B , ^
Plants, and vines on non-crop lands Includ'n9/l^e°'f ,J2 to 3 ga1/acre
frcatiins suggest Tordon 101 appHcat.on a t th. ate of J/2 ^
for the control of broad leaf weeds, and I to H gai/a
and vines.
The effect of Tordon 101 on fish has been revised by Kenaga 0 969) .
*« '"" l LC50,S for
™/L (a.e.) 2,*-
» Fathead
TtaVh LC50 for
3.7
e
s were exposed to varying level? f
»ater at 10°C. Lynn (1965 in Kenags 19«)
brook, brown, and rainbow trout were 50.3,
0, and ,3.7. ,3.1 and 86 m9/L
45
-------
Experimental Results
Static exposure —
of
and
K
"
ac iviv of th am
activity of the gills, but this lowered
^°xicitY test in December indicated a 24-h LC50
(concentratl°n of 2,4-0 and picloram combined),
G a Steep mortality curve (Fig. 7). Tordon
reduction in the (Na,K)-stimu?ated ATPase
;
noied
;
ibly
did not
to affect
„
for the deaths, but no abnormal conditions were
TABLE 17.
ATPASE °F DARLING COHO SALMON
101 'N FRESHWATER AND THE
Concentration
(mg/L)
nominal m«
Control
0.25
0.50
1.0
5.0
15.0
30.0
60.0
^ •_i___^i ^*
0.29
0.50- 0.62
1.32- 1.59
6.42- 7.35
18.1- 19.8
31.5
67.8
'
— TWeiltU fj Qh ovrnne*^y7 «
Percent
survival^/
(144-h
exposure FW)
100
100
100
100
100
100
«0/
Gill
ATPase^/
— — — — — —
1-9
NT!/
NT
1 .4
1.5
0.9
Percent
survival
(360-h SW)
100
25
65
100
100
100
Gill
ATPase£/_
4.0
4.9
5.0
4.0
,ATPase
|/Mean of 5 fish ^ea^ater
tgflot measured.
-All fish dead within 60 h.
-'All fish dead within 2.5 h
46
-------
_J
1
tr
(/)
h-
2
UJ
O
o:
UJ
CL
100
90
80
70
60
50
40
30
20
10
0
0-15 mg/L
—X X X X X
TeOmg/L]
x—x—
)Q~X X X—X- —X X X—X X
"K
\ \
\
'*. 0.5mg/L
"^.
•
\
\ 0.25 mg/L
W
^~^—.-^)_.^._.—..
48
24 72 120
Freshwater
EXPOSURE TIME (hours)
96 144
Seawater
192 240 360
=
transfer to seawater.
Another test with Tordon 101 was 'tate in
test tanks were dosed with toxicant stock that was nn erf when the
the other tanks were dosed with toxicant stock tha t P J |n toxicant
experiment was initiated. There appear ed to ^ no d Jhe
effect regardless of when the tox1"?* S5S J3/L) i«d In the March test
highest toxicant exposure concentration U^ mg/u; however, 95^ of
was c,ose to the estimated 2A-h LC50 of Decemb. (^/^..^ n-
the yearling coho salmon exposed to 22 m9^ f Tordon 101 is unknown,
tration) died. The reason for ^V^LlJearl ing coho salmon are more
but if the measured values are real . tnen ^ ' J no mortality noted
l 5W,roups represenUnHHe, of
-------
TABLE 18. SURVIVAL OF YEARLING COHO SALMON EXPOSED TO
TORDON 101 IN FRESHWATER, AND THE SUBSEQUENT
SURVIVAL FOLLOV/ING TRANSFER TO SEAWATER
(MAR. 13-29, 1977).
Concentration
(mg/L)
nominal measured
Percent
survival—/
(144-exposure FW)
Percent
survival
(240-h SW)
New stock daily
Control
0,25
0.50
1.0
22.0
0.13-0.16
0.12-0.13
8.41-8.83
Stock mixed Mar. 13. 1977
100
100
100
100
5
100
100
100
100
0
0.25
0.50
22.0
0.19-0.38
0.1*1-0.50
9.21-9.7
100
100
0
100
100
Twenty fish exposed per concentration.
Flow-through exposure--
Following marking (fin excision) and time for acclimatization to new
surroundings, yearling coho salmon were exposed to Tordon 101 in the flow-
through system beginning on Mar. 28, 1977. Groups of yearling coho salmon
were placed in seawater following 144-h and 380-h exposure to the four
Tordon 101 concentrations. No deaths occurred in either the fish exposed
S I? ?", ^TK1" ?u6 S?ntro1 fUh dUrlng the 24° h of seawater exposure
(Table A-2). The (Na,K)-st,mulated ATPase activity was not affected by any
^r6 ^°H 101 execrations tested (Table A-2). Exposure to Tordon
101 for 360 h produced no apparent effect on the condition factor of
yearling coho salmon (Table A-3). The fish all fed welt during the
exposure, and the general decline in condition factor was probably the
normal change in the length-weight relationship that occurs as the fish
undergo transformation to smolts (Wagner 1974a).
A release of 10 groups of coho salmon exposed to Tordon 101 for 360 h
and 5 groups exposed for 96 h was made into Crooked Creek on April 13,
1977, and downstream movement was monitored. The coho salmon exposed to
0.3 mg/L for 15 days showed slightly stimulated migration compared to the
controls, whereas the other concentrations (0.6, 1.2, and 1.8 mg/L) showed
a dose-dependent inhibitory response. Approximately a 10% difference in
movement was noted between controls and the group receiving 1.8 rog/L
Tordon 101 over the 84 days that migration was monitored (Fig. 8a). The
differences observed, however, were not statistically significant (P = 0.05)
-------
Coho salmon that received 36 h exposure to Tordon 101 (0.3-1.8 mg/L) prior-
to release did not migrate as well as the control group except for the
0.3 mg/L group which again showed slight migratory stimulation. In the
otherVh exposure groups (0.6, 1.2, and 1.8 mg/L) the m,fl™^™^!*
was less than observed for the control but no relat.onsh.p to concentration
of Tordon 101 was apparent (Fig. 8b). The majority of f^"'^ *?™*nt
of all groups of coho salmon occurred within the f.rst 5 days of release
(Table A-4).
(.Appendix I I). me consequences ui L...^ ^i- ; -
effect on the fish in terms of ultimate survival is unknown.
Coho salmon that had been exposed for 15 days to 18 mg/L Tordon 101
, c -.——fc—rti -rich were ki I leu ana sevciai ui j^u*. «••
i comparable group or control IISN "»= TnrHnn 10.1 No
^ • j -,,,4 onalw^^H for bioconcentrat ion or iuruun i«i. «•
, systems excised and a?a^° [°Joram was noted. Youngson and Meikle
,.././r,1owever!adidnfi>nd residues of 0.21 mg/kg in mosquitofish exposed to
1 mg/L picloram for 18 days.
DINOSEB (PREMERGE)
Review of Literature;
selective
TheY
Dinitrophenols are the oldest organic chemical patented
weed control (Kaufman 1976). Two of these c°?P°""ds' J' ^ c
A, 6-dinitrophenol) and dinosam have become widely used herb^c
Phenol form of dinoseb is oil soluble and ^T^ed ^^ suiu_ _
concentrate. The amine or *^*™s**"a^s. DOW Premerge (alkanol amine
are formulated under a variety of trade names dinitroamines and
salt) was utilized in this study. The 9enerai seedling weeds and
ammonium salts has been for postemergent control of most ^ »f ^
grasses in cereal, tree, and vegetable crops. HPP pe (g jj depending
amine salt of dinoseb is normally from •:» to g controi ditch bank
upon the crop. Dinoseb is also applied in some areas to
weeds (Woodward 1976).
i _ A. . ._.^*v*t *H n A I f"
The fate and behavior of herbicides
rate of decomposition, volatilization, m
(Kaufman 1976). Dinitrophenols fPPefrJ; Izat:lon „, WM1.
undergo photolysis at alkaline pH. voiar':!j of son acidity, high
dinoseb has been shown to occur under conait. ^ fo1iowing applicatt
temperature, and surface soil moisture. ne altnough in mineral
may cause leaching, particularly in aj.k^1'"®. (Montgomery et al. 1976,
days, adsorption may slow the rate ot |ea~n' *.., degrade dinitrophenols.
Kaufman 1976). Certain soil mi5roorga"'S^rsTsoi 1 conditions is 3-5
The residual life of dinoseb under warm, moist so ^ season to the
weeks; residual carry-over is not expected to occur
ne*t (KUngman and Ashton 19751-
n acid solutions, but
or codisti Ilation of
°
-------
70
60
50
40
30
_ 20
v°
O^
10
o
A. 360 h.
-•-o // <50,6mg/liter
l,2mg/liter-
.8mg/liter
AU.O my/1"
4 Control .
^l.8mg/IH
-a l,2mg/liteir
oo.6mg/l«ter-
0
7
y "I 13 15" 25
DAYS: POST RELEASE
31
group following 360 h of Tordon o
sal.cn released per group fdloJng
50
-------
Dinoseb has a high toxicity to -r.animals anc, fish.
chronic effects of dinoseb on cutth!:0?* *™s^gely dependent upon water
Woodward (1976), who found that toxicity was ^rgely ^P lnjrease ,n
quality. Decreasing the water pH resulted '" * ^™P™ by Llpschuetz and
dinoseb toxicity to fish. Similar findings wer; reP°rt* £H fProm 8.0 to
Cooper (1961) for technical grade d.noseb. Decreasing P ^ ^
6.9 increased the toxicity of dinoseb to "^^.J^,^ the toxicity
High water temperature and h.gh hardness also tena c Woodward
of dinoseb to fish, but to a lesser extent than pH (Webb
1976).
^ ,, and
TU nc. u ircn'e nnHpr varvinQ conditions 01 H"» t'-"^*-1 -- ' >
The 9b-h LCpU s, unaer vaiyiy dinoseb for cutthroat trout ana
hardness, ranged from 0.41 to 1.35 mg/ 1976). Lipschuetz and
from 0.032 to 1.40 mg/L for lake trout ^wo° values for rainbow trout.
Cooper (1961) observed similar dinoseb toxicity ^ ^ pR 6>9 wgs
Their 24-h LC50 at PH 8.0 was 0.30 mg/L ain°^ ] atratulus; in water
0.073 mg/L (18°C). Western blacknose dace
-------
<
>
>
o:
Z>
h-
«P
2.
QI
uj
Q_
0-75jjg/L 0-IOOjug/L
100 HT& — 8^ — K X X X
11 ^^*\ _s\
QnL\ "QjOO^g/L
i V \
80 1- i200^g/L ;
70r 1 ;*
60 1 1 6
P ' '>
50| ^ \
40 tBOOV '.
1 •! Jfll/ 1 T^ '
1 1 ^y • t
30 -W 1 \(
OO — ll^***^ 1 '
L'lJQ/Lt *
10 - ^. ! 6
Q )L x i i i
; x — x x x x x x x « x
i i i i i i i i i i
24 72 120 24 72 120 168 216 240
Freshwater Seawater
EXPOSURE TIME (hours)
Figure 9. Percent survival of yearling coho salmon during exposure to
dinoseb (Dow Premerge) in freshwater and subsequent survival upon transfer
to seawater.
TABLE 19. SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO DINOSEB IN FRESHWATER AND SUBSEQUENT SURVIVAL FOLLOWING
TRANSFER TO SEAWATER (FEB. 22-MAR. 10, 1977).
Concentration Percent
(yg/L) survival^/ Gill
nominal measured (144-h exposure FW) ATPase^/
Control
25
50
75
100
200
300
500
0
1.1- 12.0
3.0- 11.8
3.1- 13.1
16.6- 61.7
23.0- 65. k
1A1. 0-209.0
241.0-275.2
100
100
100
100
10
0
0
0
2.9
2.7
2.8
3.7,
NTS/
-
-
Percent
survival
(260-h SW)
100
100
100
100
100
-
-
—'Twenty fish exposed per concentration.
2/(Na,K)-stimulated ATPase activity; mean of 4 fish.
£/Not measured.
52
-------
Flow-through exposure —
As dinoseb was found to be extremely toxic to coho salmon and had the
potential for interacting with anadromous salmon because of its use in weed
control along ditches and irrigation canals, it was chosen for study in the
flow-through system. Following marking (fin excision) and a 5~day accli-
matization period, yearling coho salmon were exposed to dinoseb (Dow-
Premerge) beginning April 19, 1977. Two days after initiation of the
exposure, the fish in the highest concentration (60 ug/L) began to turn
dark, eat poorly, and die. The deaths occurred at concentrations con-
siderably lower than had previously caused death under static conditions in
late February. Whether the smelting of the fish, the use of a flow-through
system or a combination of both was responsible for the greater toxicity is
unknown. During the first 6 days of exposure to dinoseb, 33% mortality
occurred in the groups receiving 60 yg/L and 8% mortality was noted in the
40-yg/L group. Two deaths occurred during the sea water challenge in the
group originally exposed to 60 yg/L dinoseb for 144 h (Table 20). Con-
sidering the poor physical appearance of the 40 and 60 ug/L exposed groups
prior to placing in salt water, it was surprising that there were so few
deaths (Table 20). The majority of dark colored fish regained their
silvery coloration during the 10-day seawater challenge. There were no
mortalities during either toxicant exposure or seawater challenge of the
coho salmon exposed to 10 or 20 yg/L dinoseb for 16 days. However, all
the fish exposed to 60 yg/L and almost all exposed to 40 ug/L died during
the 16 days of exposure (Table 20). The gill (Na.K)-stimulated ATPase
activity was unaffected after 6 or 16 day exposures to dinoseb (Table 20).
TABLE 20. SURVIVAL OF YEARLING COHO SALMON EXPOSED TO DINOSEB
IN FRESHWATER AND SUBSEQUENT SURVIVAL FOLLOWING
TRANSFER TO SEAWATER (APR. ig-MAY 16, 1977).
Concentration
(yg/L)
nominal measured
Percent
survi val£/
Gill
ATPase^/
Percent
survival^/
(240-h SW) •
A. 144-h exposure
B
d
T^
Control
10
20
40
60
0.5
7-4- 7.
15.3-17-
38.6
54.5-58.
9
3
3
100
100
100
92
7
(270)
(250)
(250)
(300)
(339)
6.
as
(10)
NT-X
6.
5.
54
36
(10)
(10)
100
100
100
100
90
.0
.0
.0
.0
.0
(20)
(20)
(20)
(20)
(20)
. 384 -h exposure
Control
10
20
40
60
/Number of
0.5
6.2- 8.
15.3-17.
30.7-38.
52.9-60.
8
3
6
0
100
100
100
6
0
(270)
(250)
(250)
(300)^
(339)£
6.
NT
6.
NT
81
43
(20)
(20)
100
100
100
NT
.0
.0
.0
(30)
(30)
(39)
fish exposed in parentheses.
-f (Na,K) -stimulated ATPase
activity
/wean with
sample
size in
parentheses.
Q Not measured.
r 'P^T»w»f ns4-ax? s~*n x^an 1 ^ •^•x-* -r*^*e?-t-s~\s-tTr- •^^»»"il^^
£/All dead within 11 d.
53
-------
Crooked C "k 4 ?97
groups monitored c«h«
sHghtly better miration T
no sV nlf i^ rd: e °e n mi a
groups (Fig IDA) Of th mi9rat
control and'2 ?y A d no ebTre iel
the 10-and *0-yg/L groups had onlv
group exposed to 60 yg/L for 2?^
tendency than the controls
int°
h
though
so
™* mae n°
the downstrea^ migration of the various
^ tO 10 P9/L dlnoseb for '* daYs showed
0'5 °r the 2° ^ 9r°Up' *
imes were noted for the three
exp°8ed f°r % h' t
mi9ratlon>
o (Fig' 10B)'
g lo) n h J°* 'f^ m'^ra°
dinoseb for A8 h showed a H^l H i' Sh exP°sed to ^0 or 60 ug/L
downstream migratfon (Fig 10^ PC±«H ln|:ibitlon ln ave^ percent
occurred, however, between reo ir^H 1e Variation in migration rates
60 yg/L (Table A-5)l and on?v 2 e>
-------
_!__'• i i i i I I I I I I I I ft-
T. •=>•• inn of vearling cono &diinw»
Figure 10. Percent downstream migration vi 7 203"2I9 coho salmon
exposure to dinoseb (Dow Premerge). A. reP^S® osure. B. represents 98-115
released per group following 360 h of dinos K seb exposure except for
coho salmon released per group following 96 h o ^ ,37 coho sa mon
the 60 pg/L group exposed for 25 h. .J'^PJo ug/L dinoseb, respectively.
released following 48 h exposure to <»u ana o
55
-------
TABLE 21. ACUTE TOXICITY OF OIQUAT TO VARIOUS FISH SPECIES-7
cn
Species
Carasstus auratus
(goldfish)
Esox lucius
(northern pike)
Fundulus s imi 1 is
(killifish)
Ictalurus punctatus
(channel catfish- fry)
1 . punctatus^
Lepomfs cyanellus
(green sunfish)
L. macroch I rus
(bluegi Il-fry)
L. macrochi rus
L. macroch f rus
(f ingerl ings)
L. macrochi rus
L. macrochi rus
L. macrochi rus
Mfcropterus dolomieui
(smal f mouth bass)
M. dolomieu!
M. sal mo Ides
{largemouth bass-fry)
M. sal mo ides
H. salmon ides
Mo rone saxatl 1 is
(striped bass-tarvae)
(f ingerl Ings)
(fry)
(finger lings)
Toxfcity test
static, field
concrete pond
static, field
concrete pond
flowing, lab.
static, lab.
static, lab.
static, plastic
pool
static, lab.
static, lab.
static lab.
static, field
static, lab.
static, lab.
static, plastic
pool
static lab.
static, lab.
static, lab.
static, lab.
static, lab.
static, lab
static, lab
static, lab.
Lf SO2/
(mg/Ll pH
35 (96 h) 8.2
16 (96 h) 8.2
1.0 NTE (48 h)^7'^7
10.0 NTE (111 h)-7 8.4
10.0 NTE (96 h)-7
4.0 NTE-7 8.3-8.9
10.0 NTE (12 d)
4.0 NTE (96 h)
25.0 (30 h) 8.4
19 (48 h)
525 (24 h) 6.9
150 (48 h)
35.0 (96 h) 8.2
9-10 LD,n (97 h)-7
72 (96 h) 7.4
140 (96 h) S.I
0.5 NTE-7 8.3-8.9
4.0 LC|5 (96 h)
10.0 LD)0 (96 h)
2.5 (24 h)
1.0 NTE (96 h)
7.8 (96 h) 8.3
11.0 (48 h)
1.0 (24-96 h)
35.0 (24 h)
10.0 (96 h)
315.0 (24 h)
80.0 (9b h) 8.2
Temperature
48-89°F
48-89°F
23.3-25-8°C
75°F
22-25°C
23.3-25.8°C
24°C
25°C
48-89°F
75°F
25°C
25°C
75°F
22-25°C
22-23°C
25°C
70°F
70°F
21°C
Alkalinity Hardness
(mg/L as CaCO^) Reference
233 230 Gilderhus (1967)
233 280 Gilderhus (1967)
Butler (1965)
75 78 Jones (1962)
Lawrence et al. (1965)
120-189 Yeo (1967)
Hiltibran (196?)
75 78 Jones (1962)
Cope (1966)
40 29 Hughes and Davis (1962)
233 280 Gilderhus (1967)
Lawrence et al. (1965)
22 22
3'2 341 Surber and Pickering
(1962)
120-209 Yeo (1967)
Lawrence et al . (1965)
Hiltibran (1967)
(1962)
(197.)
Hughes (1973)
64 35 Wellborn (1969)
-------
Species
Oncorhynchus kisutch
(coho satmon-yearl ings)
0. tshawytscha
(chinook salmon)
0. tshawytscha
Pimephales promelas
(fathead minnow)
P. promelas
Rasbora hetermorpha
(harlequin fish)
Salmo gairdneri
(rainbow trout )_
S. gairdneri
S. gairdneri
S. gairdneri
S. gairdneri
S. gairdner!
S^ trutta
(brown trout-f ingerlings)
S. trutta
Stizostedion vitreum
(walleye)
Toxicity test
static, lab.
static, lab.
static, lab.
static, lab.
static, lab
static, lab
flowing, lab.
static, lab.
flowing, lab
static, lab
static, field
concrete pond
static, lab.
flowing, lab.
static, lab.
static, lab.
static, lab.
static, lab.
LC 50^/
(mg/L) pH
30 (96 h) 7.4-7.6
29.5 (24 h) 7.4-7.7
28.5 (48 h)
29.0 (48 h)
10.0 NTE (96 h)-7
14 (96 h) 7.4
130 (96 h) 8.2
73-93 (48 h)
5.0 LD)0 (96 h)-7
70 (48 h)
10 NTE-7
11.2 (96 h) 8.2
20.0 (48 h)
10.0 NTE^7 8'°
32.6 (24 h)
20.4 (96 h) 7.5
300 (48 h)|7 7.6-8.0
570 (48 h)^7
2.1 (96 h) 8.2
Temperature
10°C
20°C
75°F
25°C
25°C
20°C
65°F
20°C
55°F
48-89°F
13°C
18.3°C
10 °C
48-89°F
Alkal inity Hardness
(mg/L as CaCOi) Reference
79-82 100-101 Lorz et al. (this
study)
41-71 Bond et al. (I960)
Mui rhead-Thompson
(197D
Lawrence et al. (1965)
22 22 Surber and Pickering
299 379 (1962)
20 Alabaster (1969)
Lawrence et al . (1965)
250 Alabaster (1969)
U.S. Fish 6 Wild.
Service (1963)
233 280 Gi'lderhus (19&7)
Cope (1966)
89.5 Folmar (1976)
100 Simonin 6 Skea (1977)
210-290 Woodiwiss & Fretwell
(1974)
233 280 Gilderhus (1967)
U Concentration of diguat causing 50% mortality of exposed fish in time given
-£l NTE - Wo toxic effect.
5^ Test carried out in seatrater.
j, Approximate threshold toxicity LD1Q for 96 h contact period
Concentration of Aquacide and Reglone given as 48 H LC50 values, respectively.
-------
LcS'S \5n mudpat:tlcles • Woodiwiss and Fretwel 1 (1974) estimated a W-h
LC50 of 300 mg/L d.quat for brown trout in relatively hard water; however,
(1^77 of°7n Hhan/|te?-fold h'9her than that given by Simonin and Skea
rllllrrh rl £ M , ! ? Wat^ ab°Ut One-ha1f as hard. The majority of the
researchers (Table 21) noted that salmonids had a 96-h LC50 of 11-32 mg/L
d.quat in moderately hard water. Bond et al . (1960) reported a A8-h LC50
tL PA h^rcn T! ™P Cl]ln00k Salmon whereas in °ur study, we estimated
>2Q ™*. ?2 K 2° m?/L f°r C°h° Salmon" We did n°t test concent ratio*
steeoer »nT ^ * ^ °f the toxicitY curve may be considerably
nd^t.H ?h 9'Ve a Jrer U5° Value" Surber and Pickering (1962)
ndicated he importance of hardness, noting that the 96-h LC50 of diquat
fathLd «• WaS 5 J? 10 tlmeS the l6Vel observed 1" ^ft water tests with
Tathead minnows and bluegill sunfish.
ucean n ^ be tOXic tO fish frV' Bluegill, lake chub
d?auat und ^ -° 5"? frV surviv^ for 3 days or less in 2.5 mg/L
that rainbow f at '%cond! tlon- (Hlltlbran 1967). Folmar (1976) reported
diquat? Y d'Splayed no avoidance of 0.1, ,.0 or 10 mg/L
treatment 0^ Chickahominy Reservoir,
^h ^ i" dIqUat accum"1ation in the hydrosol .
adsorbed to
erib, h d or
voided soon aftor If Hiea neroicide is believed to be metabolized or
-- -al - 1965,
.
r
dqua (Cab^e ?0 nd ^hadd|tlona, fish were exposed to
suggested that diquat moved primarilv in th' T measured radloact. v. ty
predominantly in the aastro-lni I V plasma and accumulated
water decreased the 1 bj eS b cIS^ inT* P1aC6ment °f fiSh ln ^f"
not in the gastro-intestinal t act and i Vanous /T"5 3nd tlSSUeS> 5" h,
freshwater. ' nd increased the radioactivity of the
Experimental Results
Static exposure--
nqanth urr tO yearlin9 coho salmon was
static diquat exposure te s ^re olac ^ ™ ^ "^ When ^^^
dose dependent manner in !u P . ln seawater, deaths occurred in a
trations greater !San o ii/?"??8 °!,yearlln9 coho exposed tb concen-
size was small there w^ no ( '9< "' Tab1e 22)' Although our sample
stimulated AT sel"w ^^rnf^ °f dlqU9t °n the {Na'K)'
58
-------
3
100
90
80
70
60
50
40
30
20
10
0
rt n ^^
24
72 120 24 72 120
Freshwater Seawater
EXPOSURE TIME (hours)
11. Percent survival of year, ing.coho
<"quat In freshwater and subsequent surv.val upon transfer
Concentration
(mg/L)
£22lnal measured
0,
1,
0.23
.47-0
.05-1
4.94
9.45
14.7
19.4-19.7
49
06
Percent
survival^/
H44-h exposure FW)
100
100
100
100
100
100
70
42
GI11 h/
ATPaseg/
5.0 (4)
NT
NT
4.9 (5)
NT
4.5 (6)
NT
NT
Percent
survival
240-h SW)
100
100
100
100
85
57
36
13
-—•
«t» fish exposed per °°™ent*atl°n'. mean with sample size in parentheses.
,K)-stimulated ATPase activity of giu, » ,
59
-------
Flow-through exposure--
Additional tests were conducted with diquat because it is used as a
prophylactic treatment in fish culture and for control of aquatic vege-
tation, and affects seawater survival of coho smolts. Following tank
acclimatization, 150 yearling coho salmon per tank were exposed to diquat
starting May 13, 1977. After 144 h and 312 h of diquat exposure, groups of
yearling coho were transferred to seawater. Only one fish died (from the
highest toxicant concentration 3 mg/L for 312 h) during the seawater
challenge (Table 23). The toxicant concentrations of our composite water
samples were close to the desired concentrations. No apparent effect of
diquat on the (Na,K)-stimulated ATPase activity was noted (Table 23).
TABLE 23. SURVIVAL AND GILL ATPASE OF YEARLING COHO SALMON EXPOSED
TO DIQUAT IN FRESHWATER AND THE SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER.
Concentration
(mg/L)
nominal measured
Percent
survival^/
Gill ATPase^/
Percent
survival
(268-h S\>0
A. 144-h exposure
Control 0
0.5 0.58
1.0 1.07
2.0 2.04
3.0 3.06
B. 312-h exposure
Control
0.5
1.0
2.0
3.0
0
0.44-0.58
0.92-1.04
1.91-2.04
2.86-3.06
100
100
100
100
100
100
100
100
100
100
4.93 (20)
5.83 (20)
A.28 (20)
100 (21)2/
100 (19)
100 (20)
100 (20)
100 (20)
(240-h SW)
100 (20)
100 (29)
100 (20)
100 (31)
95 (19)
of fish exposed per concentration. ^
— (Na,K) -stimulated ATPase activity nf n-m . ™ •_..
parentheses. «• cavity of gill; mean with sample size in
^/Number of fish tested in parentheses.
n all
f°i
S
/h *™& n°
(Tab1e A"7) '
aPPare^ effect
G«nerally, the
on the condition
fish appeared
both the acu and chron cx osu res
H res
h 1 teSted' ^
The coho salmon exposed
60
-------
A. 285(1
0.5 mg/liter
,0 mg/ liter
2.0 mg/ liter
3.0 mg/liter
II 13 (5 19 23 27
Control
Ml " H
,0 mg/liter
0.5 mg/liter.
3.0 ma/liter
2O mg/iiter
I 13 (5 19 23 27
DAYS; POST RELEASE
Figure 12. Percent downstream migration of yearling coho salmon following
exposure to diquat. A. represents 178-183 coho salmon released per group
following 285 h of diquat exposure. B. represents 98-105 coho salmon
released per group following 96 h of diquat exposure.
61
-------
for 285^ showed a dose-dependent inhibition of their migration. Less
than 30% of the fish exposed to 3 mg/L diquat migrated the 6.4 km down-
stream compared to 63% of the controls. The 1, 2, and 3 mg/L groups all
showed significantly less migration than the controls (P = 0.05). In
the 96-h exposure groups the percent migration varied from k&% (2 mg/L)
to 12% (controls), but the group responses were not dose-dependent. The
0.5, 2 and 3 mg/L (96-h exposure) groups all exhibited significantly
less migration than the controls. The controls showed the best migratory
response following both acute and chronic diquat exposure (Table A-8) .
Histological examination of diquat-exposed fish showed degenerative
necrosis of the liver, kidneys, and gill lamellae of chronically exposed
coho salmon (Appendix II). |n acute exposures, however gill tissues
exhibited the most degeneration although the liver did show limited
necrosis (Appendix II).
No bioconcentration of diquat was found in muscle, kidney, or liver
tissue, although difficulty was encountered in the extraction procedure.
The literature generally indicates that diquat may enter a fish's body
^cumulated since the herbicide is metabolized or excreted
io™aPPeurS fr°m the Water (Be^ley et al. 19&5, Calderbank
I f 6Ver' Gi1derhus (1967) found diquat residues in
6 weeks after treatment with 1.0 ppm. Calderbank 1972,
al:°972) d BerrV et al. (1975) showed that most if not
a
in tish r.,ii-,,ro f * d'qUat is USed for both ^atic weed control and
in iibn culture tor t t-a^t-m^^. 4- ~£ i^^ ,_••,.,. ».«_
bacterial gill disease, further investi"
gation appers wr a 9 sease' urther nves
exposure ^or In H information concerning the effect of reduced
eva uaHon „? H °t reC°Very foll^'n9 exposure would permit a
id? f ^ c: n^Uat S P°SSible ad— e effects where migrating
62
-------
SECTION VI
GENERAL DISCUSSION
h We attested to determine the * h-LCSO value of 12 -^soluble
herbicides to yearling coho salmon. Th« ^c.ty o ft P^ ^
tested varied from <0.1 to >200 mg/L. /crolem and ,vel .
most toxic, with 96-h LC50 values of 6*™*™™'fa (dimethyl amine)
ammonium ethyl carbamylphosphonate (Krenitej ana , Several
were apparently non-toxic at concentrations of 200 mg/L. ^
researchers (Hughes and Davis 1963, Meehan et al. ™£ H tempera-
1976) have discussed the "-PPortance of alka1.n.ty, hardnes , P ,
ture, and especially chemical formulation of the herb.c.de,
Parameters related to observed toxicity to nsnes.
Most fish species, but particularly salmonids would succumb to
concentrations of acrolein or dinoseb if eJP^?a"7)> When coho salmon
application rates (Oregon Weed Control Handbook W/) seawater, few
Yearlings were removed from either herbicide and placed ^.^ Qf
additional deaths occurred. The (Na K) stimu atea
the gii]s appeared unaffected by e.ther herb.c.de.
The toxicity of some compounds may be empnas,— — --
chronic exposure. Sprague_ (1969 andEaton U9/ > Jh tQ adequateiy
tance of maintaining experimental exposures '°9 tencc of dinoseb
define lethal threshold concentrations, me H habitat contamination.
increases the possibility of continuous aqu*ntrations of 2»0 and 60 ug/L
lr> our dinoseb flow-through exposures, cone ^f exposure;
resulted in almost total mortality during tne ^ ^ g days cause(J no
whereas, in an earlier static experiment, '' ^ulative mortalities in
mortality. Woodward (1976) did not ooserv ^^ g dgys> simnarly,
cutthroat trout or lake trout exposed to d.no ^ ^ ^ djd nQt cause
'n our study concentrations of 10 and zu My
Acute exposure of yearling coho salmon to amirru, ^, subsequent
Paraquat produced moderate mortality ^_'y d t response, even at
challenge with seawater elicited a f05^^"?. No statistically ^ _
concentrations that were sublethal in rre t|muiatcd ATPase activit
significant effect of the herbicides °" ^j ' d chronically exposed to
°f the gill was observed. Coho salmon """'^ ,nto a small coastal
diquat exhibited migratory inhibition upon release
stream.
. f £ _ JK
Atrazine, Krenite, 2,4-D, and 2,*,5 ^_ _^
°n smolting of yearling coho salmon, i.e. K)_stimuiated ATPase activity.
seawater challenge test, and no effect on vna,
63
-------
mg/L)?
the
exposure
and
'"
101 (plclora.
salon preosl ov the " "^^ ° *"'""*
cide. No effect on the ?Na K? *• ? St concentrat Ion of the herbi-
was noted although f sh ^K)-stimul«ed ATPase activity of the gills
water were not always checked?0"06"^3'10"5 C3USing ^tallty in sea-
did elicit a reduced^atorvT' ^^l the tw° hi^est concentrations
days at 1.2 and 1.8 mg/L) as esp°nse of chronically exposed fish (15
exposure groups. The conceni-0?9 tO contro1 or low Tordon 101
inhibitory effect on migratio ,°ns T°rdon 101 that produced the
one would expect to see uti 1 iz'd ' ^f' are considerably greater than
Picloram is extremely persistent In/ '^,d °Peration. As noted earlier,
following application. 'Stent and could be present for up to a year
Generally, except for th H'
dinoseb and effects of subletLi ^^ tOX'C effect of acrolein and
and seawater survival, the herb' C0.ncerntrat!ons of diquat on migration
to affect smolting of yearlina r°h formulations tested did not appear
water tolerance, (Na,K)-stimulated°A?P ^ ^ as measured by sea-
n our study, however, We test^H i activity or migratory tendency-
1'fe history; whereas genetics VnY °n? ]]fe Sta9e of the salmonids'
may be considerably more affect ^ 'f& Stages' or food resources
study with atrazine. rrected as noted by Macek et al. (19?6b) In a
The insecticides DDT anH
to modify salmonid physiology S, !°n
exposure to sublethal concent rattn
Peterson 1969> Elson et al 972 u^f
and Johansen 1972, Hatfleld and Ri K \
1969, Symons 1973, 1977, Warner e at
Whether the herbicides testeTwouti i
•nsecticides is unknown. t doe,
precautions be taken during the
sheds containing f]sh. ng the
(feni trothion) have been shown
lear"ing ability following
n '971' Anderso" and
a"d Anderson 1972, Hatfield
KeenleV^de 1967 Saunders
and Wi1dish et ^- W]*> '
resP°ns« similar to these
h°WeVer> that appropriate
ion of any herbicide to water-
-------
SECTION VII
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PP
ln: P' C« Barney and D. D.
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24:808-822
Y°Ung
»* ^ DDT in
antic salmon. J. Fish. Res. Board Can.
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the
20:3243-3245.
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biphenyls: effect of l-
Plmephales promelas. Bull
insecticides. Biochem. Pharnacol.
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-6 °n ATPase activity in fish,
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70
-------
, J. R, 19^7. DNBP. pp. 385~396. _!TK_ Gunter Zweig (ed.) Analytical
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71
-------
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-------
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73
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' *"-;
of
18. HTIS PB-2351.57. 90 pp.
) acetic
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** .. Pond waters,
on the
SWi1d' S=r- ReP- No. FWS-LR-/'*
-------
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75
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76
-------
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trout. Trans. Am. Fish
Trans. lilis Acad Sc 65
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°f
*"*
Hughes, J. S. and J. T. Davis 1Q62 M H-
parts per million of bluealll tmf" k!" J°'france ^mlts reported in
-M9S2. La. wild. andVuh."^ ^^'^ %** •""•"••
78
-------
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Bull. Envir. Contam. andToxicol. 12(l):76-80.
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Contam. andToxicol. 17(l):l-l8.
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Schultz, D. P., and E. W. Whitney. 197*. Monitoring 2,4-D residues at
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146-152.
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fishes and its effectiveness in the control of emergent species of pond
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Walker, C. R. 1971. The toxicological effects of herbicides and weed
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Appl. Ecol. 8:509-532.
Yokote, M., S. Kimura, H. Kumada and Y. Matida. 1976. Effects of some
herbicides applied in the forest to the freshwater fishes and other
aquatic organisms. IV. Experiments on the assessment of acute and
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Fish. Res. Lab. (Tovko) 26(2):85-93-
79
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APPENDIX I. METHODOLOGY FOR ANALYSIS OF SPECIFIC HERBICIDES.
AMITROLE-T
1. Summary of method: The method is an adaptation of Southerland
(1964). Amlnotriazole is removed from water by adsorption on an
ion exchange resin. The eluate from the resin is decolorized with
activated charcoal. N-(1-Naphthyl) ethylenediamine dihydrochloride
is used to form a chromophoric moiety which is measured in a
colorimeter at 460 nmJ
2. Reagents: Dowex 50W-X8
3-amino-s-triazole
Sulfuric Acid, reagent grade (concentrated, 75% and 0.75%)
Sodium nitrite, reagent grade (0.5% aqueous solution,
prepare fresh daily)
Sulfamic acid, reagent grade (5-0% aqueous solution,
prepare fresh daily)
N-(l-Naphthyl) ethylenediamine dihydrochloride,
reagent grade (1.0% aqueous solution, prepare
fresh daily)
Charcoal, activiated ("Darco" G-60)
Ammonium hydroxide, reagent grade, 2 Normal so.lution
3. Apparatus: Widemouth plastic bottles, 500 mL (sample containers)
Magnetic stirrers and stir bars
Chromatographic columns
Hot plates
Colorimeter
Timer
Suction flask (500 mL)
Sintered glass funnels (medium porosity)
Beakers (600, 400 & 100 mL)
Erlenmeyer flasks (50 mL)
Volumetric flasks (25 ml)
Pi pet
Glass beads (3 mm)
81
-------
l\. Procedure:
a.
Resin adsorption and desorption of aminotriazole: Measure 200 ml of
?5S^^
beaker infn th» ? ™i J'™* the Sample and resi" fr°™ 'he
' -'
beaker infn th»
5±i,'3£ s-'s ra^^'^-i,^0^ .r^s
reSi" """ 2°° mL °f 2N TOnlura MrexlS I nto a MO mL
beaker
'' £;r"S= ?s zxxsfnsxisii s.
with
SSI's" : ~
» • ~ «•
c. Color development: Transfer a 5 ml al Inuni- nf , -
erlenmeyer, add 3 mL concentr=,,-^ ,'lc^ot: ot sample into a 50 mL
bath. Add 10 drops ofTS sodium I"''" * 9nd C°o1 'n an ice
add 10 drops of 5.0% sulfamic 'd ^lte! and swirl; after 10 min,
drops 1.0% N-(l-Naphthyl) ethylened'- ^i* Aftet" ]° m'n add 10
Read against a reagent blank at i*An ^min® °!hydrochlor1de and swirl.
standard curve for aminotMazoU d ? W!thln 30 min. Using a
triazole in the 5 mL aHquo? determine the amount of 3-aml.no-s-
ATRAZINE
1. Summary of method: This method is ha<;Pd « 7 •
Atrazine is extracted from water with ™of-h i 9Land Sherma ^972a).
carried out using a chloride specific ml/ "? Chl°rlde' A"^ysls is
graphic system. spec.tic microcoulometric gas chromato-
2. Reagents: Methylene chloride, reagent grade
Methanol, reagent grade
3- Apparatus: Glass jars with teflon-lined caps
rt^.r!'°7 fV"nels (500 mL)
Volumetric flasks
, or
halogen ti7r;;i;;UceiriCr°COUl0metriC detector
82
-------
a.
Procedure:
Extraction: Measure AOO ml of water sample into 500 ml separatory
funnel fitted with teflon stopcock. Shake sample with three successive
75 mL portions of methylene chloride combining the three portions in
a 250 mL beaker. Evaporate the methylene chloride to near dryness
using a stream of nitrogen. Add 20 mL methanol and evaporate to
near dryness again to ensure all methylene chloride is removed.
Transfer to volumetric flask with methanol.
Gas Chromatography: Gas chromatograph parameters:
Coulometer settings: 250 ft, 250 mV bias, low gain mode.
Column:
Temperatures
Column
Inlet
Transfer 1ine
Combustion furnace:
Gas flows:
Recorder:
Retention time of
atrazine:
4 mm I.D. x 1.2 m glass packed with 5%
OV-1 on 80/100 Gas Chrom Q.
175°C
205°C
225°C
800°C
Carrier gas 60 ml nitrogen/min,
oxygen 50 mL/min.
Sweep gas 20 mL nitrogen/min.
0.1 mV/in (0.04 mV/cm), 1/2 in/min
(1.27 cm/min) chart speed. Equipped
with a disc integrator.
2.8 min.
The amount of atrazine in the samples is determined by injecting an
aliquot of the methanol solution into the gas chromatograph. Compare
peak chart areas with areas for known amounts of atrazine.
DICAMBA, PICLORAM, 2,4-D, and 2,4,5-T
Residues in water
'. Summary of method: Samples are collected and stabilized with sodium
hydroxide. The samples are adjusted to a pH <2, extracted with
ether and alkylated to form the methyl ester. Detection and quantification
>s accomplished using microcoulometric gas chromatography (MCGC) in
the halogen mode.
83
-------
Reagents:
3. Apparatus
Diethyl ether (distilled over sodium)
Sulfuric acid (concentrated, reagent)
Sodium hydroxide (reagent)
Diazomethane
BF^-methanol
Widemouth plastic bottles (500 ml)
Beakers^ fU"ne1S f'tted W'th Tefl°n st°Pcocks
Volumetric flasks
Graduated cyl inder
Gas chromatograph - MicroTek 2000 MF, or equivalent
with Dohrman model C200B microcoulometric detector
with halogen titration cell.
Procedure:
us 2
laboratory and stored at
*
--
transP°>-ted to the
:L2°:,stor sample to
Extract with three successive alinnn, f ^""^ated sulfuric acid.
ether extracts are col lee ed in a '250 m°L \^ °°0' 8°' 6° mL) ' The
less than 20 ml. The ether llv.r u ^*r' 9nd evaP°>-ated to
water and into a 30 mL beJke * Thl r^-f " W deC3nted 9WaY from anY
three small aliquots of "he 'addino th ^^ ls rinsed with
extract is evaporated to 2 ml wl th I ^ rt™["3* to the beaker. The
concentrated extract Is transferrrf ^ll*™ °f dr"Y nitro9^n. This
a 10 ml volumetric flask carlfuHv H *™" aUqU°tS °f ether lnto
evaporated to dryness on'alialh evap'^or?9 "^ *"**
and 2,4,5-T may be formed
or a BF3-methanol reagent
Gas chromatography:
Gas chromatography parameters:
Coulometer settings:
Co 1 umn:
are formed
6Sters of 2'^D
1370)
n yen m\, k.
w, 250 mV bias, low gain mode.
-------
Tempe/atures
Column
Inlet
Transfer line:
Combustion furnace:
Gas flow:
Recorder:
Retention time of
methyl esters:
-T, and picloram
200°C for 2,4-D, 2
185°C for dicamba.
205°C
225°C
825°C
Carrier gas 60 ml ni trogen/min,
Oxygen 50 mL/min.
Sweep gas 21 ml ni trogen/min.
0.1 mV/in (O.OkmM/cm) 1/2 in/min
(1.27 cm/min), chart speed. Equipped
with a disc integrator.
dicamba
2,4-D
2.4,5-T
picloram
2.0 min (185°C)
2.0 min (200°C)
3.0 min (200°C)
5.5 min (200°C)
The amount of herbicide in the samples is determined by injecting an
aliquot of sample solution in the gas chromatograph and comparing
peak chart areas with areas for known amounts of the herbicide
injected.
PICLORAM, 2,4-D, AND 2,4,5~T
.Residues in Fish
urn
Summary of method: Whole body fish samples are digested with potass.
hydroxide and centrifuged. The supernate is adjusted to pH <2,
extracted with ether and alkylated to form the methyl ester. Detection
and quantification is accomplished using microcoulometric gas chromato-
graphy (MCGC) in the halogen mode.
Reagents:
Apparatus:
i n
Diethyl ether (distilled over sodium)
Sulfuric acid (concentrated, reagent)
Potassium hydroxide (reagent)
Diazomethane
95% ethanol
Separatory funnels fitted with teflon stopcocks
Round-bottom boiling flask (250 ml)
Condenser (West, 2hAO 5)
Volumetric flask, 10 ml
85
-------
Graduated cylinder, 100 ml
Heating mantle, 250 ml
Centrifuge (International Model U or equivalent)
Centrifuge bottles, 250 ml
Beakers
Flash evaporator (CaLab Model 5101 or equivalent)
Tissue grinder (Tek Mar Model SDT or equivalent)
Gas chromatograph - MicroTek 2000 MF, or equivalent,
with Dohrmann model C200B microcoulometric
detector with halogen titration cell.
Procedure:
a. Sample extraction: Weigh sample, cut in small pieces and place in
250 ml round-bottom boiling flask. Add 50 ml of 35% ethanol, grind
with tissue grinder. Add 100 ml 50% potassium hydroxide in water
solution, place in a heating mantle and attach a reflux condenser.
Reflux for one hour. Cool, transfer contents to centrifuge bottle
and centrifuge at 2000 rpm for 20 min. Decant into separatory
funnel, acidffy to
-------
**• Procedure:
a. Sample collection: Water samples are collected in 500 mL, wide-
mouth, glass bottles which contain 3 mL of 50% sodium hydroxide.
The samples are mixed to ensure complete hydrolysis of any DNBP
which may be present. The samples are transported to the laboratory
and stored at 2°C.
b. Sample extraction: Transfer 200 mL of water sample to a 500 mL
separatory funnel. Adjust to pH < 2 with concentrated sulfuric
acid. The acidified sample is extracted with three successive
aliquots of ether (100, 80, 60 mL). The ether extracts are collected
in a 250 mL beaker, evaporated to less than 20 mL. The ether layer
is carefully decanted away from any water into a 30 mL beaker. The
residual extract is rinsed with three small aliquots of ether adding
the rinsings to the beaker. The extract is evaporated to 2 mL with
a stream of dry nitrogen. This concentrated extract is transferred
with small aliquots of ether into a 10 mL volumetric flask, decanting
away any water and evaporated to dryness on a flash evaporator.
c. Derivation: The methyl ester of DNBP is formed using an ethereal
diazomethane reagent (Hopps 1970).
d. Gas chromatography: Gas chromatograph parameters:
Column: A mm I.D. x 1.2 m glass, packed with equal
amounts of 6% 0V 210 and b% SE 30 on
80-100 Gas Chrom Q.
Temperatures
Column 180°C
Inlet 200QC
Gas flow: Carrier gas 80 mL nitrogen/min.
Recorder: 0.1 mV/in (0.04mV/cm), 1/2 in/min (l.27cm/min)
chart speed.
Retention time of '
DNBP methyl ester: 3-5 min.
The amount of DNBP in the samples is determined by injecting an
aliquot of sample solution in the gas chromatograph and comparing
peak hefghts with peak_heights for known amounts of injected DNBP.
Residues in Fish
1. Summary of Method: The method is based on Lane (1967) and Zweig and
Sherma (1972b). Whole body fish samples are digested with potassium
hydroxide and centrifuged. The supernate is adjusted to pH <2,
extracted with benezene and alkylated to form the methyl ester. The
ester is cleaned up with an alumina column. Detection and quantification
is accomplished using electron-capture gas chromatography.
87
-------
Reagents:
Apparatus
Procedure
Diethyl ether (distilled over sodium)
Benzene (Burdick & Johnson, Distilled in Glass TM)
Potassium hydroxide (reagent)
Sulfuric acid (concentrated, reagent)
Alumina, activated (Fisher A-5AO)
Diazomethane
n-Hexane (distilled over sodium)
Acetone (Burdick & Jackson, Distilled in Glass TM)
Separatory funnels fitted with teflon stopcocks
Beakers
Volumetric flasks
Graduated cylinders
Pipet, Pasteur, disposable
Heating mantle, 250 mL
Round-bottom boiling flask
Condenser (West, 2VAO 5)
Centrifuge (International Model
Centrifuge bottles, 250 mL
Tissue grinder (Tek Mar Model SDT or equivalent)
Gas chromatograph (Varian Model 1200 or equivalent
with scandium E.C. detector)
Flash evaporator (CaLab Model 5101 or equivalent)
(250 ml)
U or equivalent)
Sample extraction: Weigh sample, cut in small pieces and place in
250 ml round-bottom boiling flask. Add 50 mL of 95* ethanol , grind
with tissue grinder. Add 100 mL 50* potassium hydroxide in water
solution, p ace in a heating mantle and attach a reflux condenser.
Reflux for 1 hour Cool, transfer contents to centrifuge bottle and
centrifuge at 2000 rpm for 20 min. Decant into separatory funnel,
acidify with concentrated sulfuric acid, and extract with three 75 mL
onnrf solution of diazomethane
(Hopps 1970) drop by drop until the yellow color persists. Swirl the
J ±r9e?h ly',al!°W t°1stand 10 min' and evaporate just to dryness.
Prepare the alumina column by tamping a plug of glass wool
• nto a d.sposaole Pasteur pipet and pour in a 1-in layer of activated
Dortin3; fT1^ th^.methylat-d ^sidue to the column with two 3 mL
port.ons of hexane. Discard the hexane eluate. Elute the DNBP
mL ethyl ether- coiiect
waterhanalys?s?PhY *"* quantl f '"tion are identical with the method for
D10.UAT AND PARAQUAT
-------
able to determine concentrations of diquat by direct readings of the water
at 310 nm and for paraquat at 256 nm. Concentration was determined from a
standard curve.
-------
APPENDIX II. EFFECT OF VARIOUS HERBICIDES ON HISTOLOGY OF YEARLING COHO
SALMON. BY DR. J. D. HENDRICKS, OREGON STATE UNIVERSITY.
ACROLEIN
Control t 1M h (3 fish) - All tissues were normal.
50 yg/L acrolein. UA h (3 fish) - The livers had occasional exfoliated,
necrotic cells, but were otherwise normal. One of the kidneys was normal,
one had extensive vacuolation of the collecting duct cells, while the other
had protein precipitate and cellular debris present in Bowman's capsule and
various tubular regions. Both hypertrophy and hyperplasia were seen in the
gill epithelium of fish examined.
100 yg/L acrolein, Mt h (3 fish) - Most of the liver cells were delineated
and separate from adjacent ones, some were necrotic. All the kidneys had
considerable debris and precipitate in tubule lumens, and one kidney had
extensive necrosis of both segments of the proximal tubule. All kidneys
were engorged with blood. The gill epithelium from all fish was totally
destroyed, necrotic and sloughed. Two hearts were normal. A section of
esophagus exhibited massive necrosis of all k tunics.
AMITROLE-T
Control, l¥t h (3 fish) - There was extensive peripheral, coagulative
necros.s due to bile spillage and several foci of peribiliary necrosis
within the substance of the liver. The remaining liver cells were normal,
so I assume both of these lesions were the result of poor post-mortem
handlmg of the l.vers. K.dney and gill tissues were normal.
100 mg/L Amitrole-T, |M, h (1 fish)_- All liver cells showed either hydropic
degeneration or coagulative necrosis. The necrosis was diffuse; approxi-
mately 25/c of the parenchymal cells were affected. There was extensive
coagulative necros.s of all regions of the nephrons as well as most of the
hematopo.et.c tissue of the kidney. The lamellar epithelium of the gills
Umen^'o ^h Ta"ateJ fr°m the under1Ying pillar cells on many of the
was severe H Jh V* ^ ^^ ™ theSe 3 tissues> this treatment
was severe and the changes would be incompatible with survival.
' ChangeS in the liver wer* of the same
tvoe^Snlh''0!^60 h (3 f'Sh) ' ChangeS in the liver wer* of the same
con!lH^riMMJ PPm SXpOSure but not as extensive. One of the fish had
considerable diffuse coagulative necrosis of liver cells, but the others
• - — • ii.^,i V.&I13, UUL L11C U LI 1C I
hydropic degeneration. Kidneys exhibited similar
extens' E • h T tubules and hematopoietic tissue, but not quite as
normal i!L,i lal « L*\ C&]]S ofAthe 9Hls were hypertrophied, disrupting
treatment h, T tect"re' A1though not as severe as the previous
treatment, these t.ssue changes would probably not permit longtime survival.
90
-------
ATRAZINE (AATREX)
15 mg/L AAtrex, 140 h (3 fish)- The livers and kidneys of these fish were
normal. There was hypertrophy of the gill epithelium in two of the fish,
while the third had several large aneurisms at the base of the filaments.
DICAMBA (BANVEL D)
100 mq/L. dicamba. 1M h (5 fish) - All livers exhibited foci of peripheral
and/or peribiliary bile necrosis but were otherwise normal. Kidneys and
gills were also normal in these fish. The experimental treatment had
little if any effect on the tissues examined.
PARAQUAT
100 mg/L paraquat. 120 h (4 fish) - All the livers exhibited a low grade
hydropic degeneration, particularly in centrolobular regions and occasionally
in foci of peribiliary necrosis. One kidney was normal, the other 3 had
necrotic cells in the first and second proximal tubules. Two of the fish
had gills from which nearly all the epithelium had been sloughed; where it
remained, the epithelium was degenerate or necrotic. The other two fish had
less severe gill lesions.
ESTERON [BRUSH KILLER (2,^-D + 2,4,5'T PGBE ESTER)]
Control. 96 h (5 fry)—/- Livers and kidneys were normal on all these fish,
but the gills were similar to the experimentals with regard to chloride cell
hyptertrophy. This response may be the result of something other than the
Esteron treatment. Its significance is not clear.
J2QO uq/L Esteron. 28 h (5 fry)-/- Some peribiliary necrosis was present in
all livers and some diffuse cellular degeneration was observed in two of
them. Kidneys were normal in all fish, but gills had lesions similar to the
800 yg/L treatment, i.e. hypertrophied chloride cells and lamellae engorged
with blood.
1200 ug/L Esteron. 48 h (A fry)-/- Livers were normal except some slight
degenerative changes in one. Kidneys were again normal and gills had
similar lesions to the 2 previous treatments, i.e. hypertrophied chloride
cells and blood engorged lamellae.
Esteron. 96 h + 2k h rest. (5 fryjj/- All tissues were normal
except for foci of peribiliary necrosis in 3 livers.
Control (yearling coho salmon). 96 h (3 fish) - All tissues were normal
except for some peripheral bile necrosis on the livers.
— Steelhead fry -Big Creek Salmon Hatchery.
-------
800 ug/L Esteron % h 3 fish) - Except for peripheral bile duct necrosis
l.vers were normal in al samples. Kidneys were normal with the exception
all s hL IT TUK '?.^e Collectin9 duc< cells from two fish. The
gills had several abnormal . t.es; 2 of them had curved gill filaments the
(11* Lhrnicfc ^ epithelLUm had ^«nt hypertrophled chio'de '
were enao led wi?i hi T" f Ch Were exfoli^ed, and most of the lamellae
were engorged w.tn blood, resulting in large aneurisms on some lamellae.
TORDON 22K (PICLORAM)
'
. f°Ci °f both PTlph.r.1 •"-
5 mg/L picloram. 144 h
^m» hS!l' ".Ex^ensive degenerative changes were
Mnduced H^ P'C rn,?atUre and some s^Har to cyclo-
fiber-like strands In the c ^P?^ ' LeT ^^°^ and contained
several abnormalities including hy^rtrophy of^hT3!' WhHC 9H1S
wrmkled appearance of many of the epithelial cells *' ^ 3
TORDON 101 (2,k-Q + PICLORAM)
cells of one kidney
as though it
normal except that one had peripheral
ne a perp
thlr, IT , Vac^1es in the collecting duct
™ »**
bile ducts were affed
gills were no maf ''
live- h.d degenerate bile
Perib51iary bile necrosis. Since
duct
the
K2 mg/L Tordon 101. 180 h (4 ffsh)
fron, th. Pn,ar cels
.
due to the herbicide. Kidneys and
livers had degenerate bile duct
All kidneys and gills were
epithelial
necrosis was present
b,U necrosts Mas present
Tordon 101 caused perib!! ianecrosil th*™ a9ain norma'- An ^«'s °f
so it was assumed that It was due to the treatment"* PI"eSent in the c°ntrols
DINOSEB
and Perib!"-y bile necrosis of
otherw.se the w
and Perib!"-y bile necrosis o
were normal. otherw.se they were normal. Kidneys and gills
92
-------
JO yg/L dinoseb, 336 h (3 fish) - There was some hydropic change in one of
the livers and occasional scattered necrotic cells. The other two livers
were normal. The second segment of the proximal tubule had occasional cells
with pyknotic nuclei and first proximal segment cells contained eosinophilic
droplets. Gills were normal, tissue changes from this treatment were minor.
60 yg/L dinoseb, 168 h (6 fish) - Livers, kidneys and gills from all fish
were normal. There appeared to be no effect on these tissues.
100 ug/L dinoseb, 114 h (*t fish) - All the livers had peripheral bile necrosis
and extensive diffuse necrosis of the remaining parenchymal cells. All
regions of the kidney tubules as well as most of the hematopoietic tissue
were necrotic. The gill epithelium was totally necrotic and sloughed from
the gill lamellae. This treatment was severe and extremely toxic to the
tissues examined.
DI O.UAT
Control. 360 h (5 fish) - Livers, kidneys and gills of these fish were
no rma 1.
10 mg/L diquat, ]kk h (3 fish) - Livers showed hydropic degenerative changes
in centrolobular regions and some necrosis in one liver. Kidneys of all
three fish were normal but gills showed both limited hypertrophy and hyper-
Plasia of epithelial cells.
20 mg/L diquat. iMt h (1 fish) - This fish had extensive hydropic degeneration
and necrosis of the liver. The kidney was normal but the gills had both
hypertrophy and hyperplasia of epithelial cells.
3_mg/L diquat 360 h (5 fish) - Several foci of degenerate and occasionally
necWic cells were present in the liver. The kidneys also had numerous
degenerate and some necrotic tubule cells, particularly in the collecting
ducts. Both hypertrophy and hyperplasia of lamellar and interlamellar
epithelium were present in the gills.
93
-------
APPENDIX III. METRIC AND ENGLISH EQUIVALENTS
English-to-Metric
Metric-to-English
1 inch
1 inch3
1 foot
1 foot2
1 chain
1 acre
1 foot2/acre
1 gallon(U.S.liquid)
1 pound
Degrees Fahrenheit
2.5400 cm
16-3871 cm3
0.3048 m
0.0929 m2
20.1207 meter
0.4047 ha
0.2296 m2/ha
3-7853 L
453-5924 gms
1 centimeter
1 centimeter3
1 meter
1 meter2
1 meter
1 hectare
= 0.3937 in
= 0.0610 in3
= 3.2808 ft
= 10.7639 ft2
= 0.0497 chain
= 2.4710 ac
1 meter2/hectare = 4.3560 ft2/ac
1 1'ter = 0.2642 gallon(U.S.liqu
0.0022 pounds
id)
= 9/5 (°C) + 32 Degrees Celsius - 5/9 (°F - 32)
1 meter = 100 centimeters
1 meter = 1,000 millimeters
1 hectare = 10,000 m2
94
-------
TABLE A-l. CONCENTRATIONS OF 2,4-D AND 2,4,5'T IN STEELHEAD TROUT
FRY FOLLOWING EXPOSURE TO ESTERON BRUSH KILLER^/
Exposure concentration
(yg/L)
nominal measured
A. May 31 -June
Control
75
450
450
800
800
1200
1200
1200 (+ 48 h
800 (mortal
1200 (mortal
B. June 12-16,
Control
800
1000
1200
4, 1977
32.9-34.0
35.2-37-2
77.4
80.1
V
V
Jb/
v
clean water)^/
ities)*/
itiesW
1977
6.5-7.2
202-269
240-243
226-246
Time
(h)
96
96
96
96
48
48
48
48
48
24
24
96
96
96
96
Residue found
Wet weight of (mg/kg)
tissue (g) 2,4-D 2,4,5'T
15.27
10.30
11.48
12.20
10.65
11.07
10.75
4.06
4.49
1.32
16.78
25-11
19.6
18.85
19.98
<0.05
0.90
41.0
23.1
8.70
19-57
8.28
1.48
3.39
64.5
16.47
_
5.12
7.66
11.35
<0.06
1.92
44.4
40.8
10.6
26.28
7-81
6.61
9.26
60.4
100.5
-
13-94
24.37
25.13
^ Whole body residues.
-/Samples lost during extraction.
95
-------
TABLE A-2. SURVIVAL AND GILL ATPASE ACTIVITY OF YEARLING COHO SALMON
EXPOSED TO TORDON 101 IN FRESHWATER AND SUBSEQUENT SURVIVAL
FOLLOWING TRANSFER TO SEAWATER.
Concentration
(mg/L)£/
nominal measured
Percent
survival
Gill
ATPase^/^/
Percent
survival*!/
(240-h SW)
A. 144 h exposure (Mar. 28, 1977-Apr. 13, 1977)
Control
0.2
0.6
1.2
1.8
(220)
(220)
(220)
(220)
(220)
0.02
0.34-0.36
0.32-0.49
1.07-1.24
0.94-1.25
100
100
100
100
100
3.64 + 1.46 (19)
3.10 + 1.00 (19)
3.93 ± 1.55 (20)
100 (20)
100 (22)
100 (22)
100 (22)
100 (22)
B. 380 h exposure (Mar. 28, 1977-Apr. 25, 1977)
Control
0.3
0.6
1 .2
1 .8
(180)
(180)
(180)
(180)
(180)
0
0
0
1
1
.005
.26-0.
.50-0.
•33-1.
.23-1.
39
52
40
39
100
100
100
100
100
3.
2.
2.
3.
2.
00
54
30
06
43
+ 0.86
+ 0.99
+ 0.91
+ 1.40
+ 0.80
(19)
(19)
(20)
(20)
(20)
100
100
100
100
96.
(30)
(30)
(30)
(30)
7 (30)S/
^-'Number of fish used in parenthesis.
- (Na,K)-stimulated ATPase activity of gill; mean +_ SE; number of fish
sampled in parenthesis.
^/Exposed for 480 h to seawater as fish appeared to behave differently than
other groups. One death occurred after 400 h of exposure.
96
-------
TABLE A-3- EFFECT OF TORDON 101 EXPOSURE ON AVERAGE LENGTH, WEIGHT, AND CONDITION FACTOR.
Date
of
sample
Mar. 9, 1977
Apr. 12, 1977
Days
of
exposure
05/
0
0
0
0
0
0
0
0
0
o-7
0
15
15
15
15
15
15
15
15
Tank
No.
1
• 2
7
8
3
4
5
6
9
10
1
2
7
8
3
4
5
6
9
10
Nominal con-
centration
(mg/L)
Control
Con t ro 1
0.3
0.3
0.6
0.6
1.2
1.2
1.8
1.8
Control
Control
0.3
0.3
0.6
0.6
1.2
1.2
1.8
1.8
Fork
length
cm ± SE
15-5 +
15-1 +
14.8 4
15-2 4
14.8 4
15.2 4
15.1 +
15.4 4
14.9 +
14.9 ±
15-7 +
15.4 +
15-6 +
15-5 +
15.5 +
15.6 4
14.7 4
15.6 4
15.3 +
15-4 4
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
32
17
29
26
28
22
25
20
23
17
0.21
0.
0
0
0
0
0
0
0
0
17
19
.18
.25
.24
.16
.23
.20
.21
Weight
g ±_ SE
42.9 4- 2
39.9 + 1
37.9 + 2
40.5 + 1
37.6 4- 2
40.4 + 1
39-2 + 1
41.4 4 1
39-0 4- 1
37.4 4 1
39-1+1
37.9 4
39-9 +
39.2 4
39.6 +
40.1 4
33.5 4
41 .7 +
36.9 +
38.5 +
.42
.29
.16
.86
.10
.63
.83
.49
.85
.23
.58
.14
.64
.34
.88
.96
.18
.98
.47
• 55
1
1
Condi t
factor
KFL ±_
.118 4
.163 4
.134 4
.145 +
.125 4
.130 4
.118 4
.111 4
.156 4
.125 4
0.998 4
1
1
1
1
1
1
1
1
1
.022 4
.035 4
.043 +
.024 4
.036 4
.038 4
.067 4
.016 4
.038 4
ion
SE
0.012
0.013
0.009
0.010
0.009
0.012
0.011
0.012
0.014
0.009
0.007
0.008
0.011
0.009
0.008
0.011
0.018
0.021
0.011
0.009
—Sample size 30
—'Sample size 40
-------
TABLE A-1*. PERCENT MIGRATION (TO JULY 6, 1977) OF YEARLING COHO SALMON
RELEASED INTO A SMALL COASTAL STREAM FOLLOWING ACUTE AND
CHRONIC EXPOSURE TO TORDON 101 (RELEASED APRIL 13, 1977).
Percent migration
Nominal concentration Days post release
(mg/L) 1^5 6-10 11-20 21-30 31 +
1. Acute exposure - 96 h
Control
0.3
0.6
1.2
1.8
2. Chronic exposure - 360 h
Control
0.3
0.6
1.2
1.8
60.9
59.8
50.it
55.0
61.5
63.6
64.5
51. 3
56.0
62.4
63.6
65. 4
52.2
56.0
62.4
63.6
67.3
53.9
56.9
62.4
63.6
67-3
53-9
56.9
62. 4
46.0
48. A
42.7
31.1
30.2
47.9
52.6
45-5
37.8
34.9
48.3
54.0
47-4
39-2
39.2
48.8
54.0
48.3
39.2
39-6
48.8
54.0
48.3
39-2
39-6
-------
TABLE A-5. PERCENT MIGRATION (TO JULY 6, 1977) OF YEARLING COHO SALMON
RELEASED INTO A SMALL COASTAL STREAM FOLLOWING ACUTE AND
CHRONIC EXPOSURE TO DINOSEB (RELEASED MAY 5, 1977).
Nominal concentration
yg/L
1. Acute exposures -
Control
10
20
4o
60
2. Chronic
Control
Control
10
10
20
20
40
60
60
(115)^
(113)
( 98)
(HO),.
(105)^
exposures
(103)
(100)
(104)
(105)
(100)
(103)
(109) . ..
( &&)£/'*/
(109)£/
1-5
96 h
60.0
53-1
62.2
47-3
43.6
- 285 h
47-6
59-0
59-6
60.9
59-0
43-7
39-4
26.1
35.8
Percent migration
Days post release
6-10
63-5
55-7
66.3
51.8
52.5
52.4
65.0
65-4
68.6
64.0
49-5
53-2
35.2
49-5
11-20
66.1
55-7
67.3
54.5
54.5
54.5
65.0
68.2
71.4
66.0
50.5
55.0-
37-5
56.9
21-30
66.1
56.6
68.4
54.5
56.4
54.4
65.0
69.2
72.4
66.0
51.5
56.0
37.5
56.9
31 +
66.1
57-5
68.4
54.5
56.4
54.4
65-0
69.2
72.4
66.0
51-5
56.0
38.6
56.9
—'Number of fish released in parenthesis
—'24 h exposure
— 48 h exposure
— Some mortality occurred in the exposure tanks prior to marking and release.
99
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o
o
TABLE A-6 CONCENTRATION OF DINOSEB IN VARIOUS TISSUES OF YEARLING COHO SALMON FOLLOWING
EXPOSURE TO SUBLETHAL AND LETHAL CONCENTRATIONS OF DINOSEB^/.
Control ^ a
b
20 yg/L (384 h) a
b
60 yg/L (144 h) a
b
skin
<0.01
<0.03
<0.02
<0.03
<0.02
<0.03
Loncentrai. ion 01 uinuacu \w/ •-j .
m,,<:rlP qill spleen qal 1 bladder
<0.01
<0.09
<0.0i
<0.01
<0.01
<0.01
<0.09 <0.28 <0.11
<0.15
0.09 1.4 <0.15
(0.06) (0.3)
0.07
(0.03)
<0.08 0.61 0.77
(0.38) (0.23)
<0.09
1 i ver
<0.05
<0.29
0.40
(0.06
0.15
(0.03)
<0.07
<0.08
kidney
<0.04
<0.09
0.37
(0.05)
0.11
(0.03)
<0.04
<0.11
y
On basis of wet weight of tissue examined; to obtain an adequate tissue sample generally 3-5 fish
'vTiruesCgfVineas less than «) a number are below the detection limits; numbers in parentheses below a nu^er
indicate the detection limit.
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TABLE A-?. EFFECT OF DIQUAT EXPOSURE ON AVERAGE LENGTH, WEIGHT, AND CONDITION FACTOR.
Date
of
sample
May 10, 1977
May 26, 1977
Days
of
exposure
(£/
0
0
0
0
0
0
0
0
0
<£7
0
12
12
12
12
12
12
12
12
Tank
No.
1
2
7
8
3
4
5
6
9
10
1
2
7
8
3
4
5
6
9
10
Nominal con-
centration
(mg/L)
Control
Control
0.5
0.5
1 .0
1.0
2.0
2.0
3.0
3.0
Control
Control
0.5
0.5
1.0
1.0
2.0
2.0
3.0
3.0
Fork
length
cm + SE
16.2 + 0.16
16.8 + 0.27
16.8 + 0.31
17.3 + 0.38
16.4 + 0.24
16.9 + 0.27
16.4 + 0.25
16.9 + 0.29
16.6 + 0.25
16.9 + 0.22
16.8 + 0.20
16.5 + 0.20
17-5 + 0.28
16.9 + 0.26
16.5 + 0.16
17-0 + 0.21
17-3 + 0.25
17.1 + 0.21
16.7 + 0.23
17.1 + 0.22
Weight
9 ±. SE
42.7 + 1.35
47.1 + 2.15
48.2 + 2.84
54.2 + 3.51
44.3 + 1.91
48.7 + 2.18
46.3 + 2.01
50.3 + 2.59
46.9 + 2.25
49.6 + 1.98
47.3 + 1.67
44.8 + 1.54
53-6 + 2.90
49.2 + 2.28
45-3 + 1.59
48.9 + 1.88
52.9 + 2.63
50.7 + 1.97
46.7 + 2.11
49.6 + 1.77
Condition
factor
KFL ± SE
0.994 + 0.008
0.980 + 0.011
0.992 + 0.009
1.009 + 0.009
0.988 + 0.008
0.985 + 0.011
1.026 + 0.009
1.019 + 0.011
1.001 + 0.009
1.005 + 0.008
0.986 + 0.008
0.984 + 0.009
0.977 + 0.007
0.990 + 0.009
0.987 + 0.013
0.972 + 0.013
0.994 + 0.010
1.003 + 0.015
0.981 + 0.008
0.979 + 0.009
a/
~ Sample size 30
*/Sample size 40
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TABLE A-8. PERCENT MIGRATION (TO JULY 6,1977) OF YEARLING COHO SALMON
RELEASED INTO A SMALL COASTAL STREAM FOLLOWING ACUTE AND
CHROMIC EXPOSURE TO DtQUAT (RELEASED MAY 26, 1977).
Nominal concentration
mg/L
1. Acute exposure - 96
Control
0.5
1.0
2.0
3.0
2. Chronic exposures -
Control
0.5
1.0
2.0
3.0
Percent migration
Days podt release
1-5
h
66.0
47.6
54.5
38.8
41.0
285 h
55.2
43.2
38.7
25.6
16.9
6-10
71.8
58.1
60.4
46.9
53.0
62.3
53.9
47.5
41.7
26.4
11-20
72.8
58.1
62.4
47.9
53.0
63.4
- 55.5
48.1
42.2
26.4
21-30
72.8
58.1
62.4
47.9
53.0
63.4
55.5
48.1
42.2
26.4
31+
72.8
58. l^/
62.4
47. <*-'
53.0-7
63.4
55.5
48. 1-/
42. 2-7
26. l&
—/Migration rates significantly lower than controls (P = 0.05).
102
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-79-071
2.
•*. TITLE AND SUBTITLE
Effects of Selected Herbicides on Smelting of Coho
Salmon
3. RECIPIENT'S ACCESSION NO.
5, REPORT DATE
June 1979 issuing date
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Harold W. Lorz, Susan W. Glenn, Ronald H. Williams,
Clair M. Kunkel, Logan A. Norn's and Bobby R. Loper
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
10. PROGRAM ELEMENT NO.
Oregon Department of Fish and Wildlife
Research and Development Section, 28655 Highway 34,
Con/all is, Oregon
1BA608
11. CONTRACT/GRANT NO.
R-804283
12. SPONSORING AGENCY NAME AND ADDRESS
Corvallis Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Corvallis, OR 97330
13. TYPE OF REPORT AND PERIOD COVERED
Final 1-5-77 to 1-4-78
14. SPONSORING AGENCY CODE
EPA/600/02
15. SUPPLEMENTARY NOTES
3. o Ur r (_ t- ivi c i^ i *-v n T i^v^id^
With the technical assistance of the Pacific Northwest Forest and Range Experiment
Station, Corvallis, Oregon.
16. ABSTRACT
Static 96-h LC50 values for 12 water-soluble herbicides with yearling coho salmon in
freshwater were: acrolein 68 ug/1; dinoseb 100 ug/1; picloram 5.0-17.5 mg/1; 2,4,5-T
(tri ethyl ami ne)> 10 mg/1; atrazine> 15 mg/1; diquat 30 mg/1; amitrole-T 70 mg/1;
paraquat 76 mg/1; dicamba> 100 mg/1; and Krenite and 2,4-D (dimethylamine) > 200 mg/1.
Amitrole-T, diquat, and paraquat exposure in freshwater reduced the survival of
salmon smolts placed in seawater. Diquat also inhibited downstream migration of
smolts. Under normal field use, acrolein and dinoseb could produce mortality of all
life stages of salmonids if treated irrigation waters were released into streams
prior to herbicide inactivation. The use of diquat at recommended treatment
levels could reduce downstream migration of smolts and decrease survival in seawater.
All other herbicide formulations tested appeared to have no effect on smolting of
coho salmon.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
"Herbicides, Salmon, Animal Behavior, Animal
Migrations, Water Balance, Histology,
Pollution, Toxicology.
Ib.lDENTIFIERS/OPEN ENDED TERMS
c. COSATI Field/Group
06/A,F,T
18. DISTRIBUTION STATEMENT
Release to Public
19. SECURITY CLASS (This Report/
unclassified
21, NO. OF PAGES
116
20. SECURITY CLASS (This page)
unclassified
22. PRICE
EpA Form 2220-1 (Rev. 4-77)
103
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