EPA/600/R-96/029
                                                        January, 1995
USE OF SURROGATE          IN ASSESSING CONTAMINANT RISK
          TO ENDANGERED AND THREATENED FISHES
                   Final Report - September, 1995
                F. James Dwyer, Linda C. Sappington,
                  Denny R. Buckler, Susan B. Jones

                     National Biological Service
                       Midwest Science Center
                       4200 New Haven Road
                     Columbia, Missouri 65201
                USEPA Project No: DW14935155-01-0
                          Project Officer
                        Foster L. Mayer, Jr.
                 Environmental Research Laboratory
                     Gulf Freeze, Florida 32561
                U.S. Environmental Protection Agency
                 Office of Research and Development
                 Environmental Research Laboratory
                     Gulf Breeze,  Florida 32561
                                                              Printed on Recycled Paper

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                                      NOTICE

      This is the final report of research funded under USEPA Project No DW14935155-
01-0. Other reports from this research include the interim report "Comparative toxicity of
selected chemicals to rainbow trout and endangered salmonids" (EPA/6QO/X-29/139,
Environmental Research Laboratory,  Gulf Breeze, FL).  Mention of trade names or
commercial products does not constitute endorsement or recommendation for use.
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                                 Table of Contents
                                                                              Page

Abstract	1

Introduction	,	2

Materials and Methods

      Test Organisms	4

      Chemicals  	5

      Toxicity Tests	5

      Biochemical Evaluation	6

      Statistical Analysis	6

Results and Discussion

      Quality Control	7

      Toxicity	8

      Biochemical Evaluation	12

      Management Implications	12

Summary and Recommendations	14

Acknowledgments	 .  15

References		16

List of Tables

      Table 1.      Test organisms and sources	19

      Table 2.      Sources and percent active ingredient of
                   chemicals used in toxicity assessments	20

      Table 3.      Summary of study design for
                   coldwater species	21

                                        iii

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      Table 4.


      Table 5.


      Table 6.

      Table 7.

      Table 8.

      Table 9.

      Table 10.

      Table 11.

List of Figures.

      Figure 1.
Summary of study design for
warmwater species	22

Average water quality of reconstituted
hard water	23

Control survival	 24

Acute toxicity of carbaryl	25

Acute toxicity of copper	26

Acute toxicity of 4-nonylphenol	27

Acute toxicity of pentachlorophenol	 . 28

Acute toxicity of permethrin  	29



Test series arrangement	30
List of Appendixes.,

       Appendix 1.  Species profile	 32

       Appendix 2.  Arithmetic means	39

       Appendix 2.  Exposure pH	45

       Appendix 3.  Dissolved oxygen concentrations	52

       Appendix 4.  Acute toxicity data	59
                                         IV

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                                        Abstract

       Section 7 of the Endangered Species Act requires Federal agencies to insure that any
action authorized,  funded or carried out by them is not likely to jeopardize the continued
existence of listed species or modify their critical habitat.  The wide use of pesticides and
other commercial chemicals potentially poses a risk to endangered and threatened species
since, by definition,  the distribution of listed species is limited and further adverse effects on
these populations could lead to extinction.  Surrogate species used in toxicity assessments
must be carefully selected in order to be protective of listed  species.

        At present,  the rainbow trout is considered to be an acceptable surrogate, for
coldwater fishes.  Similarly, the fathead minnow is considered to be an acceptable surrogate
for warm water fishes.  This research  project was designed to determine the applicability of
using rainbow trout and fathead minnows as surrogate species for several endangered fishes.
Coldwater  static acute toxicity tests were conducted with rainbow trout and three listed
species of salmonids - Apache trout (Oncorhynchus apache). Lahontan cutthroat  trout
(Oncorhynchus clarki henshawi). and greenback cutthroat trout fOncorhynchus clarki
stomias).  Warm water static acute toxicity tests were conducted with fathead minnow and
two listed species of cyprinids - bonytail chub (Gila elegans) and Colorado squawfish
(Ptychocheilus lucius).  In addition, warm water static  toxicity tests were conducted with the
razorback sucker (Xyrauchen texanus). Chemicals used in these toxicity assessments  were
selected in consultation with EPA to represent different chemical classes and toxic modes  of
action.  Chemicals used in testing were: carbaryl, copper,  4-nonylphenol,  pentachlorophenol,
and permethrin.

       Results from  the current studies indicated that the standard test organisms (rainbow
trout and fathead minnows) often had a similar sensitivity to toxicant exposure as the  listed
salmonid and cyprinid species.   The fathead minnow and the razorback sucker responses
were also generally similar.  However, for 30 percent (8 of  27) of the possible
surrogate/listed species comparisons,  the standard 96-h LC50 for the listed species was lower
than the surrogate species.  After 96 h of exposure, warmwater listed species were more
sensitive than  fathead minnows 33 percent of the time. However, the listed warmwater
species were always less sensitive than rainbow trout.  Hazard assessments using rainbow
trout would be protective of the warmwater species tested in this study.  After 96 h of
exposure,  the  listed salmonids were more sensitive than the rainbow trout  for 25 percent of
the comparisons.  Environmental protection procedures usually focus on protection of
populations or communities and not specific species or individuals of a species as may be
necessary  for endangered and threatened species.  These data indicate an additional margin of
safety may need to be included to protect listed salmonid species when toxicity assessments
utilize data obtained from studies with rainbow trout.

       The tests conducted in this study were static acute toxicity tests with  six listed species.
Further testing should be conducted with other listed species or their FWS-identified
surrogate species before definitive policy decisions concerning the protection of endangered
and threatened species are made.  In addition, chronic toxicity assessments should be
conducted  in order to compare chronic responses between  listed and surrogate species.


                                            1

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                                      Introduction

       The Federal Insecticide, Fungicide and Rodenticide Act (FIFRA; PL 80-104) requires
the U.S. Environmental Protection Agency (EPA) to register all pesticides before use in the
United States.  The Toxic Substances Control Act (TSCA; PL 94-469) requires regulation of
commercial chemicals, other than pesticide products, that present a hazard to human health
or to the environment. The Clean Water Act (CWA) specifies "it is the national policy that
the discharge of toxic pollutants in toxic amounts be prohibited" (Section 101(a)(3)).

       Under FIFRA, TSCA,  and CWA, the EPA is charged with administering and
implementing these regulations and is thus responsible for determining if the manufacture,
use, or disposal of a chemical will present an unreasonable risk of injury to human health or
the environment (Rand and Petrocelli, 1985).  In addition to EPA, two agencies of the
Department of Interior have similar interests in environmental protection, the U.S. Fish and
Wildlife Service (FWS) and the National Biological Service (NBS).  The FWS has
responsibility to conserve, protect, and enhance the Nations' fish and wildlife resources and
their habitats; the NBS mission is to  gather, analyze and disseminate the information
necessary for the wise stewardship of the Nation's  natural resources, and to foster an
understanding of biological systems.

       Section 7 of the Endangered Species Act requires Federal agencies to insure that any
action authorized,  funded or carried out by them is not likely to jeopardize the continued
existence of listed species or modify  their critical habitat. The EPA a'nd the FWS  have been
cooperating to determine the effects of pesticides and other chemicals on listed species under
Section 7 of the Endangered Species  Act.  In June  1989, a Biological Opinion was completed
by the FWS which listed 165 endangered and threatened species (primarily aquatic) and
evaluated the hazard of 113 chemicals.  Additionally, in July  1989 the EPA published its
proposed Endangered Species Protection Program in the Federal Register (USFWS,  1990).

       Chemicals inevitably enter the environment  because of their wide use by agriculture,
industry, and the public.  About 70% of the 540 million kilograms of pesticides sold each
year are used in agriculture.  Furthermore, the American Chemical Society estimates over
63,000 chemicals are currently in use (Ramade, 1988). The TSCA Inventory lists over
70,000 chemicals that can be commercially produced and used.  Some of these TSCA
Inventory chemicals are produced in  amounts of billions of kilograms per year.
Approximately  14,000 chemicals are nonpolymers that are produced in quantities of over
4,530 kilograms per year.  In  1989, the EPA Toxics Release Inventory (TRI) estimated the
release of 320 chemicals  from industrial facilities into various environments (USEPA,
1991a). For these 320 chemicals alone, the TRI reported releases of 2.6 billion kilograms
into the environment by 22,650 facilities.  The itemized summary of toxics entering the
environment included: (1) 86 million kilograms released into streams, rivers, and other water
bodies; (2) 250 million kilograms transferred to Publicly Owned Treatment Works; (3) 544
million kilograms injected underground; (4) 415 million kilograms transferred to treatment

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and disposal facilities; (5) 202 million kilograms disposed of in landfills; and (6)  1,807
million kilograms released into the air.

       In order to evaluate a contaminant's potential for impact on the environment,
standardized toxicity tests are conducted using standard test organisms as surrogates for other
species.  Under FIFRA or TSCA the aquatic risk assessment process  for pesticides and other
commercial chemicals includes: (1) acute toxicity tests with freshwater, estuarine, and marine
fish and invertebrates; (2) embryo-larval and life cycle studies with fish and invertebrates; (3)
residue studies; and (4) field testing. The freshwater fishes most often used in acute toxicity
tests include one or more of the following species: rainbow trout (Qncorhynchus  mykiss).
brook trout (Salvelinus fontinalis). coho salmon (Oncorhynchus kisutch). bluegill  (Lepomis
macrochirus').  channel catfish (Ictalurus punctatus). and fathead minnows (Pimephales
promelas).

       The CWA provides an integrated approach to protection of aquatic ecosystems
through the development of water quality criteria and the control of toxic discharges
(National Pollutant Discharge System -  NPDES). Water quality criteria are derived to
protect aquatic organisms from unacceptable adverse effects.  However, there is allowance
for some adverse effect including a small reduction in survival, growth, or reproduction in
sensitive species (Stephan et al., 1985). The derivation of acute water quality criteria for
fresh water requires a minimum data base which includes:  (1) the family Salmonidae in the
class Osteichthyes; (2) a second family in the class Osteichthyes, preferably a commercially
or recreationally important warmwater species; (3) a third family in the phylum Chordata
(may be in the class Osteichthyes or may be an amphibian  etc.); (4) a planktonic  crustacean;
(5) a benthic crustacean; (6) an insect;  (7) a family in a phylum  other than Arthropoda or
Chordata; and (8) a family in any order of insect or  any phylum not already represented
(Stephan et al., 1985).  As part of the NPDES permit system, protection of freshwater
aquatic environments  from toxic discharges  commonly includes whole effluent toxicity tests
with Ceriodaphnia dubia. fathead minnows,  and algae (Selenastrum capricornutum) (USEPA,
199 Ib).

        Inherent in these programs is the assumption that the test species used for toxicity
assessments are predictive of other species,  including endangered and threatened  species.
However, the  number of species tested is always limited.  Mayer and Ellersieck (1986)
compiled an acute toxicity data base for 410 chemicals and 66 species of freshwater animals
and concluded: (1) for a given chemical, acute toxicity among species ranged  over 5 orders
of magnitude;  (2) for a given species, acute toxicity  among chemicals ranged over 9 orders
of magnitude;  and (3) no single species was always the most sensitive among chemicals.

       Surrogate species are typically organisms that are easily tested using standardized
methods.  However, these species may  not represent populations of endangered and
threatened  (listed)  species.  The wide use of pesticides and other commercial chemicals
potentially  poses a risk to endangered and threatened species since, by definition, the
distribution of listed species is limited and further adverse effects on these populations could

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lead to extinction. Listed species may not be protected, or unnecessary regulatory programs
may be implemented, if the sensitivity of listed species is not evaluated.  Therefore, the EPA
has expanded its role in protection of listed species to include determination of adverse
effects of contaminants on survival, reproduction, and food organisms (USFWS,  1990).

       At present, the rainbow trout is considered to be an acceptable surrogate for
coldwater fishes.  Similarly, the fathead minnow is considered to be an acceptable surrogate
for warmwater fishes.  This research project was designed to determine the applicability of
using rainbow trout and fathead minnows as surrogate species for several endangered fishes.
Coldwater static acute toxicity tests were conducted with rainbow trout and three listed
species of salmonids - Apache trout (Oncorhynchus apache), Lahontan cutthroat trout
fOncorhynchus clarki henshawi). and greenback cutthroat trout (Oncorhynchus claria
stomias).  Warmwater static acute  toxicity tests were conducted with fathead minnow and
two listed species of cyprinids - bonytail chub (Gila elegans) and Colorado squawfish
(Ptychocheilus lucius"). In addition, warmwater static toxicity tests were conducted with the
razorback sucker (Xyrauchen texanus).  Appendix 1 provides a brief listing profile for each
threatened or endangered  species used  in these studies.  Chemicals used in these toxicity
assessments were selected in consultation with EPA to represent different chemical classes
and toxic  modes of action. Chemicals used in testing were: carbaryl, copper, 4-nonylphenol,
pentachlorophenol, and permethrin.

       Variation in responses between species may best be determined by the use of
biochemical/physiological measurements.  Radioligand binding is a technique which measures
the number of receptors sites in a brain preparation  and  the affinity of the receptor for a
specific radioligand.  Exposure to either carbaryl or permethrin would be expected  to
increase the concentration of acetylcholine in  the brain, albeit through different mechanisms.
This increase could be expected to alter one or both of the binding parameters of the
muscarinic cholinergic receptor (MChR) in the brain. For this reason, we also conducted a
series of screening tests to evaluate the use of receptor binding as a biomonitoring technique.
                                Materials and Methods

Test organisms

       Rainbow trout, Apache trout, Lahontan cutthroat trout, greenback cutthroat trout,
bonytail chub, Colorado squawfish,  and razorback suckers were obtained from various
government and commercial sources (Table 1).  Fish were received during the spring and
summer of 1992 and 1993 as eyed-eggs at the Midwest Science Center (MSC,  formerly
National Fisheries Contaminant Research Center), NBS, Columbia, MO.  Fathead minnows
were from MSC stocks, or purchased commercially.

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       Once received at the MSC, fish were cultured in well water (alkalinity 258 mg/L as
CaCO3, hardness 286 mg/L as CaCO3, pH 7.8,) until testing began.  All fish were cultured
at 18°C, except trout were hatched and cultured until swim-up in well water chilled to 10°C.
Before the start of the toxicity tests, fish were acclimated and held for a total of 96 h
(USEPA,  1975;  ASTM, 1988).  For the first 48 h, fish were incrementally acclimated to the
test water and temperature.  Fish were then moved to clean containers and held for an
additional 48 h at the test temperature in  100% test water.  Fish were fed during the first 48
h of acclimation but not fed during the 48 h of holding.

Chemicals

       Chemicals used in testing included carbaryl, copper, 4-nonylphenol,
pentachlorophenol, and permethrin. Chemicals were selected  to represent different chemical
classes and toxic modes of action and,  with one exception, because of the existence of
historical  toxicity data.  Carbaryl is a carbamate insecticide that inhibits cholinesterase
activity.  Copper occurs from mining, industrial applications, and in fungicide formulations.
Copper interferes with osmoregulation.  Pentachlorophenol is a  chlorinated phenol and is
used as a  wood preservative and molluscicide.  Pentachlorophenol is an uncoupler of
oxidative phosphorylation.  Permethrin is a pyrethroid insecticide and causes neurotoxicity.
Nonylphenol was selected because of a lack of toxicity data and its widespread use in the
manufacture of nonylphenol ethoxylate detergents.  Nonylphenol is a monoalkyal phenol
which may cause narcosis and may act as  an oxidative stressor.  Sources for the chemicals
and percent purity are listed in Table 2.  Organic chemical stocks were prepared by
dissolving the chemical in reagent grade acetone, while copper was dissolved in deionized
water.  Complete analytical confirmation of stock concentrations is yet to be completed.
Maximum volume of acetone added to  any test container was 7.5 mL (0.5 mL/L).

Toxicity tests

       Static acute toxicity tests  were conducted in basic accordance with EPA (USEPA,
1975) and the  American Society  for Testing and Materials (ASTM, 1988) procedures during
the summer and  fall of 1992 and 1993.  Fish exposures were conducted in 19.6 L glass jars
containing 15 L  of test solution.   All tests with salmonids were  conducted at 12°C and tests
with cyprinids and the catastomid were conducted at 22°C.  Test water was reconstituted hard
water (alkalinity 110-120 mg/L as CaCO3, hardness 160-180 mg/L as CaCO3).  Water
quality (alkalinity, hardness, and pH) was measured on each batch of reconstituted water.
Dissolved oxygen was measured  on the control, low, medium, and high exposure
concentrations at 0 h, and in those same treatments if fish survived, at 48 and 96 h of
exposure.  Additionally, pH was measured on the control, low,  medium, and high exposure
concentrations at 0 h, and in those same treatments if fish survived, at 96 h of exposure.
Tests were conducted under ambient lighting.

       A test series consisted of six exposure concentrations (three replicates per
concentration) with a  60% dilution factor. Two years of toxicity tests were conducted with

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each listed species (except greenback cutthroat trout), and each listed species test had a
corresponding surrogate species test.  For rainbow trout and fathead minnows Runs, 1-4
were conducted in 1992 and Runs 5-6 were conducted during 1993.  For all other species,
Run 1  was conducted during  1992 and Run 2 was conducted during  1993.  Both a solvent
control and a dilution water control were included for each  species (three replicates for each
combination).   Individual test series were randomly assigned to a waterbath and location
within a waterbath (complete block design - Fig 1).

       Fish were counted into groups of five with two groups pooled for each exposure
replicate (10 fish/jar - 30 fish/exposure concentration).  Fish mortality was the biological
end-point observed at 3, 6, 9, 12, 18, 24, 48, 72, and 96 h of exposure. Mortality was
defined as the lack of movement for a 5-s observation with  the unaided eye.  Dead fish  were
removed at each observational time.  The study design is summarized in Table 3 and 4.

Biochemical evaluation

       Muscarinic cholinergic receptor binding evaluations were done on brains from
rainbow trout,  Lahontan cutthroat trout, fathead minnows, and razorback suckers that
survived  the 96-h acute exposure  to carbaryl and permethrin.  Brains of fish from each
treatment were removed and pooled to obtain adequate tissue.  Radioligand binding was
measured using [3H]N-methyl scopolamine ([3H]NMS) according to the method described by
Bylund et al. (1982).

Statistical analysis

       The LC50 and 95% confidence interval for each test was usually calculated using
probit analysis.  However,  when probit analysis was not appropriate, LCSOs and confidence
intervals  were calculated using moving average, untrimmed  Spearman-Karber, or a non-
linear interpolative procedure (Stephan, 1977).

       Distribution of LCSOs usually can not be tested for normality due to an insufficient
number of LC50 estimates. Thus, it is suggested that a geometric mean be used to stabilize
the variation (Snedecor & Cochran, 1980) for determining the central tendency of a group of
LCSOs. Sappington  (1995) determined, from this same research,  that the replicate 96-h
LC50's for rainbow  trout and fathead  minnows were normally distributed, therefore  an
arithmetic mean was reported. However, there were insufficient replicates to  test for
normality for the listed species.  For this report, the replicates were  pooled within a Run and
the pooled LCSOs were used to calculate a geometric mean for each  chemical and species.
Arithmetic means are summarized in Appendix 2.

       Analysis of variance and least square difference mean separations were determined on
ranked LCSOs at 12, 24, and  96 h of exposure.  In order to summarize the 12, 24, and 96 h
responses,  the geometric mean LC50 was determined for each particular
chemical/species/time combination.  Differences between the surrogate species and listed

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species were evaluated separately for coldwater and warmwater species.  Only those tests for
which an LC50 could be calculated were used for statistical analysis.  Biological Opinions
written as a result of consultations held under Section 7 of the Endangered Species Act,
usually estimate the number of individuals likely to be "taken" as a result of an action
(USFWS, 1994).  However, most environmental assessment procedures estimate effects on
populations rather than individuals of a population.  Since a greater degree of protection is
required for endangered species (protection of individuals rather than a population), it is
necessary to minimize Type 2 errors (acceptance of the null hypothesis that LCSOs are the
same, when they are different).  For this reason, differences were considered significant at
p_<_0.1.  However, the differences at p_<.0.05 are also presented.
                                 Results and Discussion

Quality control

       Water quality for each batch of reconstituted hard water was within acceptable ranges
for alkalinity and hardness, but average pH was above 8.0 (Table 5).  Test chemicals spiked
into the test water did not affect the pH. Appendix 3 lists the 0 and 96 h  pH range,  which
represents the lowest and highest pH measured for all exposure concentrations and replicates
within a test.  Appendix 4 is a list of the exposure replicates and concentrations for which
dissolved oxygen was below  60% saturation at 48 h of exposure or below  40% saturation at
96 h of exposure.  Overall, dissolved oxygen drops were isolated  and interspersed throughout
the exposures.  However, for Run 2 and 4 of  the rainbow trout exposures  there was  a trend
for dissolved oxygen to fall below the percent saturation limits. These two Runs of fish had
the largest average weight.  The LCSOs did not appear to be different for these two Runs or
for any other test in which low dissolved oxygen was measured. Therefore, no tests were
eliminated from the statistical analysis due  to dissolved oxygen falling below percent
saturation limits.

       Organic chemical stocks from tests conducted during 1992 were analyzed to confirm
nominal concentrations.  Chemical analysis was conducted at Mississippi State Chemical
Laboratory (Mississippi State, MS) by gas  chromatography.  The average  percent of nominal
concentration and  spike recovery were:  carbaryl - 93% (n=5), spike recovery 91%;  4-
nonylphenol - 99% (n=7), spike recovery 74%; pentachlorophenol - 79%  (n=7), spike
recovery 91%; and permethrin 93%  (n=5), spike recovery 91%.   One permethrin sample
had a percent nominal concentration  of 308%, however the biological results  from the tests
using this stock were no different than  the tests conducted with the other permethrin  stocks
used in toxicity tests during  1992 and 1993 (n=8).  For this reason we believe the reported
value for this sample was incorrect.  Therefore, it was not included in the average percent of
nominal calculation.

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       Organic chemical stocks from tests conducted during 1993 were also analyzed at
Mississippi State Chemical Laboratory to confirm nominal concentrations.  The average
percent of nominal concentration and spike recovery were: carbaryl - 119% (n=3), spike
recovery 83%; 4-nonylphenol - 160% (n=3), spike recovery 103%; pentachlorophenol -
174% (n=2),  spike recovery 58%; and permethrin 128%  (n=3), spike recovery 88%.  One
pentachlorophenol sample had a percent nominal concentration of 572%,  however the
biological results from the tests using this stock were no different than the tests conducted
with the other pentachlorophenol stocks  used in either  1992 or 1993 (n=9). For this reason
we believe the reported value for this sample was incorrect. Therefore, it was not included
in the average percent of nominal calculation.

       Copper stocks were confirmed at the MSC by atomic absorption spectrophotometry.
The average percent of nominal concentration for copper was: 1992 - 100% (n=7); and 1993
- 96%  (n=4).  Spike recovery was 98%.

       Control survival, regardless of species,  with and without solvent,  was always greater
than  90% (Table 6).

Toxicity

       All LC50 calculations are based on nominal concentrations.  Appendix 5 is  a
complete listing of all the toxicity data (LC50,  confidence interval,  and slope) for each
chemical, species, Run (pooled replicates), and time period.

       In general, after 96 h of exposure, permethrin was the most toxic and carbaryl was
the least toxic. The two phenolic compounds (4-nonylphenol and pentachlorophenol)
exhibited similar toxicity.  For the coldwater species, the order of toxicity from most to least
toxic was permethrin  > copper > pentachlorophenol _>. 4-nonylphenol >  carbaryl. For the
warmwater, species the order of toxicity from most to  least toxic was permethrin >
pentachlorophenol  >_ 4-nonylphenol > copper > carbaryl. The rainbow  trout 96-h LC50
values  were lower than the fathead minnow LC50s for the five chemicals tested (Table  7 -
11).  Macek and McAllister (1970) exposed 12 species of fish (five families) to nine
insecticides and  determined that salmonids were the most sensitive and that cyprinids and
ictalurids were the least sensitive. Mayer and Ellersieck (1986)  compared the sensitivity of
four  fish families to 65 chemicals and found that salmonids were the most sensitive and
cyprinids the least sensitive. Results from our  study are consistent with their findings.

Carbaryl

       Fish exposed to higher concentrations of carbaryl were quickly immobilized.  Fish
dying from carbaryl exposure generally exhibited arched backs, gaping mouths, and flared
gills  and fins.

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       The Apache trout and Lahontan cutthroat trout were significantly more sensitive to
carbaryl exposure (LCSOs - 3.29 and 4.38 mg/L, respectively) than the rainbow trout at 12 h
of exposure (LC50 - 6.76 mg/L; Table 7).  The greenback cutthroat trout was less sensitive
(LC50 - 8.50 mg/L) than the rainbow trout at 12 h of exposure.  The Apache trout was also
more sensitive (LC50 - 2.50 mg/L) than the rainbow trout (LC50 - 4.04 mg/L) at 24 h of
exposure.  At 96 h of exposure, there was no significant difference in  sensitivity between any
of the coldwater species with LCSOs ranging  from  1.54 mg/L for the Apache trout to 2.25
mg/L for the Lahontan  cutthroat trout.

       The warm water  fishes exhibited a different time relationship (Table 7).  While there
were no significant differences in sensitivity at 12 and 24 h of exposure between the
warmwater species, at 96 h of exposure the LCSOs for the bonytail chub (3.49 mg/L) and
Colorado squawfish (3.07 mg/L) were significantly lower than the surrogate fathead minnow
(5.21 mg/L). Observational analysis indicates that while the bonytail chub and Colorado
squawfish were more sensitive than fathead minnows, they were  no more sensitive than  the
rainbow trout.

       Beyers et al. (1994) exposed bonytail  chub and Colorado  squawfish to carbaryl in
static renewal tests using 26 d old squawfish  and 6-d-old chub.  The median lethal
concentration in their tests was 1.31 mg/L for the Colorado squawfish  and 2.02 mg/L for the
bonytail chub.  Their results are similar to our findings under static conditions with larger
fish.

       The LCSOs for rainbow trout and fathead minnows from the current studies with
carbaryl (Table 7) are similar to reported values calculated from  static  toxicity tests.  Mayer
and Ellersieck (1986) reported 96-h "LCSOs for rainbow trout exposed to carbaryl ranging
between <0.32 to 3.5 mg/L with a geometric mean of 1.60 mg/L (n = 15). Marking et al.
(1984) reported LCSOs  (n=5) ranging between 0.94 and 1.74 mg/L for rainbow trout
exposed to carbaryl.  Reported 96-h LCSOs for fathead minnows range from 7.7 to 14.6
mg/L (Mayer and Ellersieck, 1986; n=3) and 6.7 to 12.0 mg/L  (Henderson et al., 1960;
n=2).

Copper

       The sensitivity of the listed coldwater species to copper was not significantly different
than the surrogate rainbow trout at  12, 24,  or 96 h of exposure.  The LCSOs at 96 h ranged
from 0.07 mg/L for the Apache and Lahontan trout to 0.08 mg/L for the rainbow trout
(Table 8).

       At 96 h  of exposure, both the bonytail chub (LC50 - 0.22 mg/L) and the razorback
sucker (LC50 - 0.27 mg/L) were more sensitive than the fathead minnow (LC50 -  0,47
mg/L).  The bonytail was also more sensitive than  the fathead  minnow at 12 and 24 h of
exposure.  LCSOs for the bonytail were 0.30 (12 h) and  0.24 mg/L (24 h) compared to
LCSOs for the fathead minnow of 1.30 (12  h) and 0.73 mg/L (24 h).  However, as was the

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case for carbaryl, both the surrogate and listed warm water species were less sensitive than
the rainbow trout.

       The 96-h LC50s for copper in the current studies are similar to reported values.
Mayer and Ellersieck (1986) report a copper LC50 of 135 /tg/L for rainbow trout (hardness -
44 mg/L as CaCO3) and 838 /zg/L for fathead minnows (hardness - 272 mg/L as CaCO3).
Calamari and Marchetti (1973) report a copper LC50 of 890 /j.g/L for rainbow trout (290
mg/L as CaCO3). Fathead minnows tested under static conditions had LC50 ranges of 22 to
1,760 /*g/L (Pickering and Henderson, 1966; hardness 20 to 360 mg/L as CaCO3). In the
Ambient Water Quality Criteria for Copper (USEPA, 1985), the Species Mean Acute Value
for rainbow trout is 42.5 /*g/L and 115.5 /*g/L for fathead minnows (hardness of 50 mg/L as
CaCO3).

4-Nonylphenol

       During the toxicity tests with 4-nonylphenol, fish exposed to higher concentrations
often exhibited increased mucus production and a white particulate material which formed on
the fins and gills.

       At 24 h of exposure, the Apache trout and Lahontan cutthroat trout were significantly
more sensitive (LC50 of 0.24 and 0.25 mg/L, respectively) to 4-nonylphenol exposure than
were the rainbow trout (LC50 of 0.30 mg/L; Table 9).  However, after 96 h of exposure, all
four coldwater species had similar LC50s (range 0.15 mg/L - greenback cutthroat trout, to
0.19 mg/L - rainbow trout).  There was no significant difference between the LC50 values
for listed warmwater species and the fathead minnow at 12, 24,  or 96 h of exposure.  The
96-h LCSOs ranged from 0.17 mg/L for the razorback sucker to 0.29 mg/L for the bonytail
chub.

       Little information is available concerning the acute toxicity of 4-nonylphenol.
However, the U.S. EPA Draft Report - Chemical Hazard Information Profile for
Nonylphenol (Etnier, 1985) - reports toxicity information from various sources.   For rainbow
trout, one 96-h LC50 was reported as 230 ^tg/L and another 96-h LC50 was reported in the
range of 560 to 920 /ig/L. The 96-h LC50 for fathead minnows was reported as 135 /ig/L
and 300 /xg/L. The reported LCSOs are greater than the LCSOs  calculated for rainbow trout
in this study, but similar to the LC50 values that we derived for fathead minnows.

Pentachlorophenol

       The Apache trout was significantly more sensitive to pentachlorophenol than was the
surrogate rainbow trout at 96 h of exposure. The 96-h LC50 for Apache trout was 0.11
mg/L while the LC50 for rainbow trout was 0.16 mg/L (Table 10). There were no
significant differences between the LCSOs at 12  and 24 h of exposure for any of the
coldwater species.
                                          10

-------
       None of the LC50s for the warmwater listed species were significantly different from
those for fathead minnows, regardless of time period. The 96-h LCSOs for the warmwater
fish ranged from 0.23 mg/L for the bonytail chub to 0,28 mg/L for the razorback sucker.

       In the current studies, LCSOs for the rainbow trout are about 3 times greater than
those previously reported by other authors, while the results with the fathead minnow are
similar to reported values.  Davis and Hoos (1975) reported that LCSOs for rainbow trout
exposed to pentachlorophenol ranged between 44 and 92  /«g/l with  a geometric mean of 68
(n=6)  for rainbow trout. Mayer and Ellersieck (1986) reported LCSOs for rainbow trout
ranging between 34 and 115 jug/L with  a geometric mean of 59 /ng/L (n=3).  These authors
also reported the acute toxicity of pentachlorophenol to fathead  minnows as 205 /*g/L. The
Ambient Water Quality Criteria for Pentachlorophenol (USEPA, 1986) reports a species
mean acute value  of 35.34 /xg/L for rainbow trout and 63.11 /ig/L  for fathead minnows.

Permethrin

       The LCSOs for Apache trout and Lahontan cutthroat trout were significantly less than
the LC50 for the surrogate rainbow trout at 24 and 96 h  (Table 11). After 24 h of exposure,
the LC50 for Apache trout and Lahontan cutthroat trout were 2.27  and 1.93 jug/L,
respectively.  The 24-h LC50 for rainbow trout was 3.78 fj.g/L. At 96 h of exposure, the
LC50 for Apache trout (1.71 /ug/L) and Lahontan cutthroat trout (1.58 /*g/L) was about half
that for rainbow trout (3.31 ng/L). There was no significant difference in the 12-h LCSOs
for the coldwater  fishes.

       The razorback sucker was significantly more sensitive to permethrin at 96 h of
exposure than was the fathead minnow. While not statistically confirmed (the LC50 was
greater than highest concentration tested), the bonytail chub was much less sensitive to
permethrin exposure than was the fathead minnow at 12,  24, and 96 h of exposure.  The
Colorado squawfish was also less sensitive to permethrin  than was  the fathead minnow.
Permethrin-exposed fish were often hyperactive - darting  throughout the water column,
tremoring, and skimming the water surface.

       Mayer and Ellersieck (1986) reported LCSOs for rainbow trout exposed to permethrin
ranging from 2.9  to 8.2 /ig/L with  a geometric mean of 5.1 /ng/L (n=9).   The LC50  for
fathead minnows was 5.7 /tig/L.  These LCSOs are  similar to those calculated in the current
study for rainbow trout and fathead minnows (Table 11).

Comparisons to other studies

       Few studies have evaluated  the sensitivity of endangered species to  contaminant
exposures.  Beleau and Bartosz (1982) conducted toxicity tests with Colorado squawfish,
humpback chub, and northern squawfish.  They exposed  these fish  to 13 inorganic and eight
organic toxicants.  The authors stated that there was a margin of safety associated with the
use of  channel catfish and fathead minnows as surrogates since  the  listed species they tested

-------
were more tolerant to the contaminants used in their toxicity assessments. However, they
made their comparison based on information reported in the literature for fathead minnows
and channel catfish. In addition, their tests were conducted in water different than that used
in the fathead minnow and channel catfish  studies.  We conducted Colorado squawfish and
fathead minnow tests concurrently and in the same standardized water.  These reasons
possibly explain differences between the current studies and those of Beleau and Bartosz
(1982).

       The current research indicated that bonytail chub and Colorado squawfish were
significantly more sensitive to carbaryl exposure than were fathead minnows, but less
sensitive than rainbow trout. This finding  is supported by work done by Beyers et al.
(1994). They conducted 4-d renewal acute tests and 32-d early life-stage toxicity tests  with
carbaryl, Sevin 4-oil, and malathion.  These authors compared their results from the 4-d
renewal acute test to acute data summarized by Mayer and Ellersieck  (1986).  They
determined that for carbaryl exposures, the Colorado squawfish and bonytail chub were
similar in response to the cutthroat trout (Oncorhynchus clarki). Atlantic salmon (Salmo
salart, rainbow trout and brook trout.  The listed species were more sensitive (2 to 10  times)
than fathead minnows, bluegill, and channel catfish.   Conversely, Colorado  squawfish and
bonytail chub exposed  to malathion were similar in sensitivity to fathead minnows and
channel catfish and less sensitive than the trout and bluegill.

Biochemical evaluation

       The number of muscarinic cholinergic receptors  (MChR) decreased in the fathead
minnow and the razorback sucker brains at carbaryl  exposures of 3.6  mg/L  and higher.
Lahontan cutthroat trout brain receptor number decreased at a carbaryl concentration of 2.2
mg/L and higher while those for rainbow trout decreased at 1.3 mg/L and higher.  Exposure
to permethrin caused a reduction in receptor number in  all four species relative to the
control.  There was no change in affinity in any of the samples.

       This preliminary study indicated a downregulation or decrease in  receptor number in
brains from fish exposed to each of these contaminants as well as a difference in MChR
binding between cold and warmwater species and between listed and surrogate species. These
results reflect a compensatory response to the increase in acetylcholine resulting from
exposure to these contaminants.  Further studies utilizing lower concentrations of
contaminant,  longer exposure periods and a recovery period would enhance  our ability to
evaluate species  sensitivity differences.

Management  Implications

       Environmental protection procedures usually focus on protection of populations  or
communities and not specific species or individuals of a species as may be necessary for
endangered and threatened species.  For this reason,  additional margins of safety may be
required.


                                           12

-------
       At 12, 24, and 96 h of exposure, there are a total of 81 possible comparisons of
LCSOs between a listed  species and surrogate species.  Exposures of the greenback cutthroat
trout to copper, pentachlorophenol, and permethrin are not included in this total since the
LCSOs for these chemicals could not be estimated.

       For the warmwater species at 12 and 24 h of exposure (30 possible comparisons), the
listed species were more sensitive twice (12 and 24-h exposure of bony tail chub to copper)
and less sensitive four times (12 and 24-h exposure of bony tail chub  and Colorado squawfish
to permethrin).  This would seem to indicate that for these listed species, evaluations of risk
for short (pulse) exposures  (e.g. water quality criteria exceedance,  agricultural pesticide
spraying,  or runoff events) could be estimated using surrogate species.  However,  at 96 h of
exposure (15 possible comparisons) the listed warmwater species were more sensitive 5 times
and less sensitive only once.

       At 12 and 24 h of exposure, the coldwater listed species were more sensitive than the
rainbow trout eight times (22 possible comparisons).  This would indicate the need for
careful evaluation of short-term exposure risk.  At 96 h of exposure  the listed species were
more sensitive three  times (12 possible comparisons).

       The 96-h LC50 is the endpoint typically used  in hazard assessment.  When  deriving  .
water quality criteria, the Criterion Maximum Concentration can be determined for important
species by dividing the Species Mean Acute Value by two (Stephan et al., 1985).   Dividing
the fathead minnow LC50 by two may not be protective of the more sensitive warmwater
listed species we tested.  However, the LC50s for the rainbow trout  were always less than
the LC50 for the listed warmwater species we tested.  National criteria typically include
rainbow trout as a test species. These criteria would probably be protective of the listed
warmwater species used  in this study.  Caution must be exercised when national water
quality criteria are modified by states in setting state water quality  standards.  This is of
particular concern when  toxicity data for sensitive species, such as rainbow trout, are
eliminated.

       As was the case  with the listed  warmwater species, dividing the rainbow trout Species
Mean Acute Value by two would not necessarily be protective of the coldwater listed species
tested in these studies.   If a national water quality criterion is close to half the rainbow trout
Species Mean Acute Value, an additional margin of safety may need to be included.

       Ecological risk procedures for endangered species and pesticides (Urban and Cook,
1985) require formal consultation if expected environmental concentrations exceed 1/20 the
LC50 (when  slopes are  not available) or 1/10 the LC10 (when slopes are available).  Results
from this  study with warmwater and coldwater species support the validity of this assessment
procedure.
                                           13

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                             Summary and Recommendations
                                                              •%

        Federal agencies need to assure that chemical use regulations comply with the
 Endangered Species Act and that these regulations are protective of threatened and
 endangered (listed) aquatic species.  Surrogate species used in toxicity assessments must be
 carefully selected in order to be protective of listed species.

        Results from the current studies indicated that the standard test organisms (rainbow
" trout and fathead minnows) often had a similar sensitivity to toxicant exposure as the listed
 salmonid and cyprinid species.  The fathead minnow and the razorback sucker responses
 were also generally similar. However, for 30 percent (8 of 27) of the of the possible
 surrogate/listed species comparisons, the standard 96-h LC50 for listed species was lower
 than for the surrogate species.  After 96 h of exposure,  warmwater listed species were more
 sensitive than the fathead minnow 33 percent of the time.  However, the listed warmwater
 species were always less sensitive than rainbow  trout.  Hazard  assessments using rainbow
 trout would be protective of the warmwater species tested in this-study.  After 96 h of
 exposure, the listed salmonids were more sensitive than the rainbow trout for 25 percent of
 the comparisons. Environmental protection procedures usually focus on protection of
 populations or communities and not specific species or individuals of a species as may be
 necessary for endangered and threatened  species. These data indicate an additional margin of
 safety may  need to be included in order to protect listed salmonid species when  toxicity
 assessments utilize data obtained from studies with rainbow trout.

        The tests conducted in this study were static acute toxicity tests with six listed species.
 Further testing should be conducted with other listed species or their FWS-identified
 surrogate species before definitive policy decisions concerning the protection of endangered
 and threatened species are made.  In addition, chronic toxicity assessments should be
 conducted in order to compare  chronic responses between listed and surrogate species.
                                            14

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                                  Acknowledgments

       The authors thank Dr. Foster L. Mayer, Jr. of the Gulf Breeze Environmental
Research Laboratory, EPA, for his guidance and assistance in this project. We thank Eugene
Greer for culturing the test organisms, and the members of the Fish and Invertebrate
Toxicology Section and the Ecotoxicology Section of the Midwest Science Center for their
assistance during this project. This project was sponsored in part by the U.S. Environmental
Protection Agency's Office of Research and Development Ecological Risk Assessment
Research Program under Interagency Agreement No. DW14935155. We  also thank ICI
Americas Inc., and Rhone-Poulenc for donating technical grade material to be used in
testing.
                                          15

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                                    REFERENCES

American Society for Testing and Materials.  1988.  Standard practice for conducting acute
       toxicity tests with fishes, macroinvertebrates, and amphibians. E 729-88.  American
       Society for Testing and Materials, Philadelphia.

Beyers, D.W., TJ. Keefe, and C.A. Carlson.  1994.  Toxicity of carbaryl and malathion to
       two federally endangered fishes, as estimated by regression and anova. Environ.
       Toxicol. Chem. 13:101.

Beleau, M.H. and J.A. Bartosz.  1982.  Part 3:  Acute toxicity of selected chemicals: Data
       Base,   In - Final Project Colorado River Fishery Project, Contracted  Studies, FWS,
       Salt Lake City, UT.  pp 242.

Bylund, D.B., J.R.  Martinez, and D.L. Pierce. 1982.  Regulation of autonomic receptors in
       rat submandibular gland.  Mol. Pharmacol. 21:27.

Clamari, D. and R. Marchetti.  1973. The toxicity of mixtures of metals and surfactants to
       rainbow trout (Salmo gairdneri Rich.).  Water Res. 7:  1453.

Davis, J.C. and R.A. Hoos.  1975.  Use of sodium pentachlorophenate and  dehydroabietic
       acid as reference toxicants for salmonid bioassays.  Jour. Fish. Res. Board Can.
       32:411.

Etnier, E.L., Chemical hazard information profile - Nonylphenol,  Draft Report,
       September, 17, 1985.  41 p.

Henderson, C., Q.H. Pickering, and C.M. Tarzwell.   1960. The toxicity of  organic
       phosphorus and chlorinated hydrocarbon insecticides to fish.  Trans.  Second  Sem.
       Biol. Problems Water Pollut. U.S. Public Health Serv., Robert A. Taft Sanit. Eng.
       Center, Cincinnati, OH. 13  p.

Macek, KJ. and W.A. McAllister.  1970.  Insecticide susceptibility of some  common fish
       family representatives.  Trans. Am.  Fish. Soc.  99:20.

Marking, L.L., T.D. Bills and J.R.  Crowther.   1984.  Effects of five diets on sensitivity of
       rainbow trout to eleven chemicals.  Prog. Fish-Cult. 46:1.

Mayer, F.L., Jr., and M.R, Ellersieck.  1986.  Manual of Acute Toxicity: Interpretation and
       Data Base for 410 Chemicals and 66 Species of Freshwater Animals.   U.S. Fish
       Wild.  Serv. Resour. Publ.  160.  579 p.
                                          16

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Pickering, Q.H. and C. Henderson.  1966. The acute toxicity of some heavy metals to
       different species of warmwater fishes.  Air. Water Pollut. Int. Jour. 10: 453.

Rand, G. and S. Petrocelli.  1985. Fundamentals of Aquatic Toxicology: Methods and
       Applications, McGraw-Hill,  Washington, D.C., 666 p.

Ramade, F.  1988. Ecotoxicology, John Wiley and Sons,  NY, 262 p.

Sappington,  L.C.  1995.  Use of surrogate species in assessing contaminant risk to
       endangered and threatened fishes.  MS Thesis. University of Missouri,
       Columbia, MO.  82 p.

Snedecor, G.W. and W.G. Cochran.  1980.  Statistical methods.  Iowa State
       University Press.  Ames, IA. 507 p.

Stephan, C.E.  1977. Methods for  calculating an LC50.  In: F.L. Mayer and J.L. Hamelink
       (eds.) Aquatic Toxicology and Hazard Evaluation, ASTM STP 634, American Society
       for Testing and Materials. 65 p.

Stephan, C.E., D.I. Mount, D.J. Hansen, J.H. Gentile, G.A. Chapman, W.A. Brungs.
       1985. Guidelines for deriving numerical national water quality criteria for the
       protection of aquatic organisms and their uses. PB85-227049.  98 p.

Urban'D.J. and Cook, NJ.   1985.  Hazard evaluation division, standard evaluation
       procedure -  Ecological risk assessment.  EPA 540/9-85-001.  U.S.  Environmental
       Protection Agency, Office of Pesticide Programs, Washington, D.C.  96 p.

U.S. Environmental Protection Agency.   1975.  Methods for acute toxicity tests with fish,
       macroinvertebrates, and amphibians.  EPA 660/3-75-009, Ecological Research Series,
       Washington, D.C. 61 p.

U.S. Environmental Protection Agency.   1985.  Ambient water quality criteria for copper -
       1984. EPA 440/5-84-031.  Office of Water Regulations and Standards Criteria and
       Standards Division, Washington,  D.C.  142 p.

U.S. Environmental Protection Agency.  1986.  Ambient  water quality criteria for
       pentachlorophenol.  EPA 440/5-86-009.  Office of Water Regulation and Standards
       Criteria and Standards Division, Washington,  D.C. 127 p.

U.S. Environmental Protection Agency.  199la.  Toxics in the Community: National and
       Local Perspectives. EPA  560/4-91-014, Office of Toxic Substances, Washington,
       D.C. 374 p., 11 appendixes.
                                          17

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U.S. Environmental Protection Agency.  199Ib. Technical support document for
       water-quality based toxics control.  EPA 505/2-90-001.  Office of Water,
       Washington, D.C.. 143 p.

U.S. Fish and Wildlife Service.  1990.  Advances in the Endangered Species/Pesticide
       Labeling Program. Endangered Species Technical Bulletin 15(12),  12 p.

U.S. Fish and Wildlife Service.  1994.  Draft - Endangered species consultation handbook.
       (730 FW 4A).  Washington, D.C.
                                          18

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Table 1.
Test organisms and sources.
Species
rainbow trout (Oncorhynchus
mykiss)
Apache trout (Oncorhynchus apache)
greenback cutthroat trout
(Oncorhvnchus clarki stomias)
Lahontan cutthroat trout
(Oncorhvnchus clarki henshawi)
fathead minnow (Pimephales
promelas)

bony tail chub (Gila elegans)
Colorado squawfish (Ptychocheilus
lucius)

razorback sucker (Xyrauchen
texanus)
Source
Beity's Enterprise
Valley, WA
Ennis National Fish
Hatchery (NFH)
Ennis, MT
Williams Creek NFH
White River, AZ
Saratoga NFH
Saratoga, WY
Lahontan NFH
Gardnerville, NV
MSC cultures
Osage Fisheries
Osage Beach, MO
Dexter NFH
Dexter, NM
Dexter NFH
Dexter, NM
Dexter NFH
Dexter, NM
Strain
Beity's
Backcross, Erwin,
and Shasta
East Fork
GCD greenback
Pyramid lake
•NA1
NA
NA .
NA
NA
'NA - not applicable
                                           19

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Table 2.      Sources and percent active ingredient of chemicals used in acute toxicity tests
              with rainbow trout and listed salmonids.
CHEMICAL
Carbaryl
Copper sulfate
4-Nonylphenol
Pentachlorophenol
Permethrin
SOURCE
Donated by Rhone-
Poulenc Agricultural
Co., Research Triangle
Park, NC
Fisher Chemical, St.
Louis, MO
Fluka Chemical, New
York, NY
Aldrich Chemical,
Milwaukee, WI
Donated by ICI Americas
Inc., Richmond, CA
ACTIVE
INGREDIENT
(%)
99.7
25.5
85.0
99.0
95.2
                                            20

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Table 3.


Test type:

Test volume:
Summary of study design for the comparative toxicity of selected chemicals to
rainbow trout and cold water species.

             Static acute

             15 L
Test temperature:

Water Quality:

Chemicals:

Species:
Exposure design:
Observations:
             12°C
             Reconstituted ASTM hard

             Carbaryl, copper, 4-nonylphenol, pentachlorophenol, permethrin

             rainbow trout - 6 Runs
             Run 1 - 0.67 ± 0.35 g
             Run 2 - 1.25 ± 0.57 g
             Run 3 - 0.27 ± 0.07 g
             Run 4 - 1.09±0.38g
             Run 5 - 0.48 ± 0.08 g
             Run 6 - 0.50 ± 0.21 g

             Apache trout - 2 runs
             Run 1 - 0.85 ±0.49 g
             Run 2-0.38 ±0.18 g

             greenback cutthroat trout - 1 run
             Run 1 -0.31 ±0.17 g

             Lahontan cutthroat trout - 2 runs
             Run 1 - 0.34 ± 0.08 g
             Run 2 .- 0.57 ± 0.23 g

             Dilution series - 60%
             Replicates - 3/concentration
             Organisms  - 10/replicate

             Mortality at 3, 6, 9,  12, 18, 24, 48, 72, and 96 h of exposure
                                          21

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Table 4.


Test type:

Test volume:
Summary of study design for the comparative toxicity of selected chemicals to
fathead minnows and listed warmwater species.

             Static acute

             15 L
Test temperature:

Water Quality:

Chemicals:

Species:
Exposure design:
Observations:
             22°C
             Reconstituted ASTM hard

             Carbaryl, copper, 4-nonylphenol, pentachlorophenol, permethrin

             fathead minnows - 5 Runs
             Run 1 - 0.32 ±0.16 g
             Run 2 -0.56±0.19g
             Run 3 - 0.45 ± 0.35 g
             Run 4 - 0.40 +. 0.21 g
             Run 5 - 0.34 ± 0.24 g
             Run 6-Q.39±0,14g

             bonytail chub - 2 runs
             Run 1 - 0.29 ± 0.08 g
             Run 2 - 0.52 ± 0.09 g

             Colorado squawfish - 2 runs
             Run 1 - 0.32 ± 0.05 g
             Run 2 - 0.34 ± 0.05 g

             razorback sucker - 2 runs
             Run 1-0.31 ±0.04g
             Run 2 - 0.32 ± 0.07 g

             Dilution series - 60%
             Replicates - 3/concentration
             Organisms - 10/replicate

             Mortality at 3, 6,  9, 12,  18, 24, 48, 72, and 96 h of exposure
                                         22

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Table 5.      Average measured water quality characteristics of reconstituted water used in
              the acute toxicity tests with coldwater and warmwater fishes.
Water Quality
Parameter
Alkalinity1
Hardness1
pH
Nominal
value
110- 120
160 - 180
7.8-8.0
Coldwater
Measured
(n =7)
117 ± 8
169 ± 10
8.24 ± 0.29
Warmwater
Measured
(n = 6)
117 ±4
m± 9
8:35 ± 0.29
'mg/L as CaC03
                                           23

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Table 6.      Average control survival (standard deviation in parentheses) for coldwater  and
              warm water species.
Species
rainbow trout
Run 1
Run 2
Run 3
Run 4
Run 5
Run 6
Apache trout
Run 1
Run 2
Lahontan cutthroat
Run 1
Run 2
greenback cutthroat
Run 1
fathead minnow
Run 1
Run 2
Run 3
Run 4
Run 5
Run 6
bonytail chub
Run 1
Run 2
Colorado squawfish
Run 1
Run 2
razorback sucker
Run 1
Run 2
Without
Solvent
100
100
100
100
100
100
100
96.8 (5.2)
100
100
96.7 (5.8)
100
100
100
100
100
100
100
100
100
100
100
100
With
Solvent
100
100
100
100
100
100
96.7(5.8)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
                                           24

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Table 7.      Acute toxicity of carbaryl (mg/L) to 8 species of fish (2 surrogate and 6 listed)
              at  12, 24, and 96 h of exposure.  Toxicity values are the geometric mean  of
              the LC50s (number of LC50s in parentheses) used  in the rank analysis. When
              significant, the probability associated with determining  whether a listed species
              is different than the surrogate is also listed (results  from analysis of variance).
              An asterisk (*) indicates a least square difference mean separation of p _<. 0.1
              and (**) indicates ap  < 0.05.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
6.76
(4)
p<.01
3.29**
(2)
8.50*
(1)
4.38**
(2)
12.0
(1)
ns1
7.93
(2)
>10.0
(NA)2
8.88
(1)
24-h LC50
(n)
4.04
(6)
p = .07
2.50**
(2)
3.59
(1)
3.60
(2)
8.25
(3)
ns1
6.13
(2)
6.31
(1)
6.67
(2)
96-h LC50
(n)
1.88
(6)
n.s.1
1.54
(2)
1.55
(1)
2.25
(2)
5.21
- (5)
p=.06
2 49**
'(2)
3.07**
(2)
4.35
(2)
'ns =  not significant
2NA = not applicable
                                            25

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Table 8.      Acute toxicity of copper (mg/L) to 8 species of fish (2 surrogate and 6 listed)
              at 12, 24, and 96 h of exposure..  Toxicity values are the geometric mean of
              the LCSOs (number of LCSOs in parentheses) used in the rank analysis.  When
              significant, the probability associated with determining whether a listed species
              is different than the surrogate is also listed (results from analysis of variance).
              An asterisk (*) indicates a least square difference mean separation of p _<. 0.1
              and (**) indicates a p  0.03
(NA)2
0.39
(2)
1.30
(4)
p=.04
0.30**
(2)
>1.00
(NA)
>1.00
(NA)
24-h LC50
(n)
0.12
(4)
ns1
0.09
(2)
>0.03
(NA)
0.11
(2)
0.73
(4)
p=.08
0.24**
(2)
0.64
(2)
0.39
(1)
96-h LC50
(n)
0.08
(4)
ns1
0.07
(1)
>0.03
(NA)
0.07
(2)
0.47
(6)
p=.05
0.22**
(2)
0.43
(2)
0.27**
(2)
'ns = not significant
2NA = not applicable
                                           26

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Table 9.      Acute toxicity of 4-nonylphenol (mg/L) to 8 species of fish (2 surrogate and 6
              listed) at 12, 24, and 96 h of exposure.  Toxicity values are the geometric
              mean of the LC50s (number of LC50s in parentheses) used in the rank
              analysis.  When significant, the probability associated with determining
              whether a listed species is different than the surrogate is also listed (results
              from analysis of variance).  An asterisk (*) indicates a least square difference
              mean separation of p _<_ 0.1  and (**) indicates a p _<_ 0.05.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
0.35
(6)
ns1
0.30
(2)
0.38
(1)
0.29
(2)
0.38
(6)
ns1
0.56
(2)
0.45
(2)
0.29
(2)
24-h LC50
(n)
0.30
(6)
p=,04
0.24**
(2)
0.30
(1)
0.25**
(2)
0.33
(6)
ns1 -
0.49
(2)
0.28
(2)
0.22
(2)
96-h LC50
(n)
0.19
(6)
ns1
0.17
(2)
0.15
(1)
0.18
(2)
0.27
(6)
ns1
0.29
(2)
0.26
(2)
0.17
(2)
'ns = not significant
                                            27

-------
Table 10.     Acute toxicity of pentachlorophenol (mg/L) to 8 species of fish (2 surrogate
              and 6 listed) at 12, 24, and 96 h of exposure.  Toxicity values are the
              geometric mean of the LC50s (number of LC50s in parentheses) used in the
              rank analysis.  When significant, the probability associated with determining
              whether a listed species is different than the surrogate is also listed (results
              from analysis of variance). An asterisk (*) indicates a least square difference
              mean separation of p _<. 0.1 and (**) indicates a p <_ 0.05.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
0.22
(6)
ns1
0.21
(2)
>0.01
(NA)1
0.27
(1)
0.33
(6)
ns1
0.42
(2)
0.23
(2)
0.53
(2)
24-h LC50
(n)
0.17
(6)
ns1
0.21
(2)
>0.01
(NA)
0.23
(2)
0.30
(6)
ns1
0.26
(2)
0.16
(2)
0.29
(2)
96-h LC50
(n)
0.16
(6)
p = .06
0.11**
(2)
>0.01
(NA)
0.17
(2)
0.25
(6)
ns1
0.23
(2)
0.24
(2)
0.28
(2)
'ns = not significant
2NA =  not applicable
                                            28

-------
Table 11.     Acute toxicity of permethrin (/xg/L) to 8 species of fish (2 surrogate and 6
              listed) at 12, 24,  and 96 h of exposure.  Toxicity values are the geometric
              mean of the LC50s (number of LCSOs in parentheses) used in the rank
              analysis.  When significant, the probability associated with determining
              whether a listed species is different than the surrogate is also listed  (results
              from analysis of variance).  An asterisk (*) indicates a least square  difference
              mean separation of p _<. 0.1 and  (**) indicates a p <. 0.05.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
5.75
(6)
ns1
3.88
(2)
>1.0
(NA)1
3.33
(2)
13.43
(4)
ns1
>25.0
(NA)
>25.0
(NA)
13.05
(1)
24-h LC50
(n)
3.78
(6)
p = .08
2.27*
(2)
>1.0
(NA)
1.93**
(2)
9.73
(5)
ns1 •
>25.0
(NA)
>25.0
(NA)
8.87
(1)
96-h LC50
(n)
3.31
(6)
p = .08
1.71*
(2)
>1.0
(NA)
1.58*
(2)
9.38
(6)
p = .08.
>25.0
(NA)
24.4
(1)
5.95*
(2)
'ns = not significant
2NA =  not applicable
                                            29

-------
CO
o
       Water
        Bath
         8
SURR.
Rep 1
                            12  Q
                            10
                 11
                  9
           Carbaryl
           Copper
           4-Nonylphenol
                                               Controls
                                              Endangered (END.)
                                              END.-Acetone
           Pentachlorophenol O Surrogate (SURR.)
       77A Permethrin       © SURR.-Acetone
              END.
              Repl
        END.;
            "H
       iRep2
        END.
        Rep1
END>
Rep3
      . SURR.
       Rep 3
                     SURR.
                     Rep 2
SURR.
Rep1
                                   SURR,
                                   Rept
           5
           6
SURR.
Rep 2
                     END.
                     Rep1
       SURR
       Rep 2
END.
Rep3
                      END,
                      Rep 3
Rep3
                      END.
                     Rep 2
SURR.
Rep 3
                                                                O4
                      Figure 1.  Test series assignment to a waterbath and location within
                              a waterbath (complete block design).

-------
APPENDIX 1. SPECIES PROFILES
             31

-------
Appendix L Species profile - Apache Trout  (Oncorhynchus
                    Historic range:  The historic range is difficult to determine because by
                    the time they were positively identified pure populations were
                    diminished.  The apparent range included the upper Salt River system
                    of Arizona and headwater streams of the little Colorado River and Blue
                    River of the San Francisco drainage.

                    Current range:  It is estimated that pure strains of Apache trout occupy
                    5% of their former range.  Those areas  include Boggy Creek, Crooked
                    Creek, South Fork Diamond Creek, East Fork of the White River,
                    Centerfire Creek, and Soldier Creek.  These streams are located on the
                    Fort Apache Indian Reservation and The Apache-Sitgreaves National
                    Forest in Arizona.

                    Reason for decline:   The reasons for population decline include;
                    reduction in  habitat, detrimental interactions with introduced species,
                    competition, predation and hybridization with cutthroat trout
                    COncorhynchus gila).

                    Status: Threatened
References

Behnke, RJ. and M. Zarn.  1976. Biology and management of threatened and endangered
       western trouts.  USDA For. Serv. Gen. Tech. Rep.  RM-28, 45 p. Rocky Mt. For.
       and Range Exp. Stn.   Fort Collins, CO.

Behnke, RJ. 1979.  Monograph of the native trouts of the genus Sal mo in western North
       America.  USDA For. Ser. Lakewood, CO.  163 p.

Johnson, I.E. and J.N. Rinne.  1982.  The Endangered Species Act and Southwest Fishes.
       Fisheries. 7:2,

Miller, R.R.  1972.  Classification of the native trouts of Arizona with the description of a
       new species, Salmo apache.  Copeia 3:401.

U.S. Fish and Wildlife Service.   1979.  Recovery plan for Arizona trout, Salmo apache.
       Miller, 1972.  Albuquerque, New Mexico. 37 p.
                                          32

-------
Appendix 1. Species profile - greenback cutthroat trout  (Oncorhynchus clarki stomias)

                    Historic range:   The historic range of the greenback cutthroat trout
                    includes the head waters- of South Platte basin and Arkansas River
                    basins in Colorado.

                    Current range:  Only three pure populations of greenback cutthroat
                    trout remain including Como Creek, South Fork - Cache La Poudre
                    River, and Cascade Creek in Colorado.

                    Reason for decline:  Declines in population were due to irrigation,
                    mining pollution, logging, grazing and introduction of exotics (brown
                    and brook trout) that displaced or hybridized with greenbacks.

                    Status:   Threatened
References

Behnke, R.J. and M. Zarn.  1976.  Biology and management of threatened and endangered
       western trouts.  USDA For. Serv. Gen. Tech. Rep. RM-28, 45 p. Rocky Mt. For.
       and Range Exp. Stn.   Fort Collins, CO.

Behnke, R.J.  1979.  Monograph of the native trouts of the genus Sal mo in western North
       America.  USDA For. Ser.  Lakewood, CO.  163 p.

U.S. Fish and Wildlife Service.  1983.  Greenback cutthroat trout recovery plan.   Denver,
       CO. 45 p.
                                          33

-------
Appendix 1. Species profile - Lahontan cutthroat trout  (Oncorhynchus clarki henshawp

                    Historic range:  The historic range of the Lahontan cutthroat trout
                    included the Lahontan basin of California and Nevada, major lacustrine
                    populations in Pyramid Lake, Walker Lake, Independence Lake and
                    Lake Tahoe in California and Summit Lake in Nevada, and fluvial
                    populations in the Carson, Walker and Truckee River systems in
                    Nevada.

                    Current range:  All populations are believed to be lost except Summit
                    Lake, Nevada and Independence Lake, California.

                    Reasons for decline:  The Pyramid Lake stock was lost due to
                    construction of Derby Dam in 1905 and Lahontan Dam (Reservoir) in
                    1915 inhibiting migration from Pyramid Lake to Truckee River.  Thus,
                    there has not been any successful reproduction since 1938,  The Lake
                    Tahoe populations declined due to damming, pollution and siltation.
                    The population in Walker Lake was reduced due to irrigation, diversion
                    and reduction of stream  flows which  interfered in reproduction.

                    Status:  Threatened
References

Behnke, R.J. and M. Zara.  1976.  Biology and management of threatened and endangered
       western trouts. USDA For. Serv. Gen. Tech. Rep. RM-28, 45 p. Rocky Mt. For.
       and Range Exp. Stn.  Fort Collins, CO.

Behnke, RJ. 1979.  Monograph of the native trouts of the genus Salmo in western North
       America.  USDA For. Ser.  Lakewood,  CO. 163 p.

Gertstung, E.R.  1988. Status, life history, and management of the Lahontan cutthroat trout.
       Am. Fish. Soc. Symposium 4:93.

Swanson, R.G.   1992.  New legislation aids the recovery of Endangered fish and refuge
       wetlands in Nevada.  Endangered Species Technical Bulletin. 7:1.
                                         34

-------
Appendix  1.  Species profile - Bonytail chub (Gila elegans)
                    Historic range:  The historic range of the bonytail chub included the
                    upper and lower basins of the Colorado River, the Green River (WY,
                    UT), Yampa and Gunnison River (CO), and the Gila and Salt River
                    (AZ).

                    Current range: The current range for the bonytail includes Lake
                    Mojave and Desolation and Cataract Canyon of the Upper Colorado
                    River basin.

                    Reason for decline:  The reasons for species decline include building
                    of impoundments, water withdrawal and the introduction of non-native
                    species.

                    Status:  Endangered
References

Carter, J.G., R.A. Valdez, R.J. Ryel and V.A. Lamarra. 1985. Fisheries habitat dynamics in
       the upper Colorado River.  J. Freshwater Ecology. 3:249.

Minckley, W.L.  1991.  Native fishes of the grand Canyon Region:  An Obituary?  Colorado
       River Ecology and Dam Management, Proceedings of a symposium.  National
       Academy Press, Washington, D.C. p.  124.

U.S. Fish and Wildlife Service. 1990.  Bonytail chub recovery plan. Denver, CO. 35 p.
                                          35

-------
Appendix 1. Species profile - Colorado squawfish (Ptychochelius lucius)

                    Historic range:  Colorado squawfish were common in the lower
                    Colorado River basin in Arizona and Colorado until the 1930s.

                    Current range:  The largest populations occur in the Green River sub-
                    basin and upper Colorado River basin.

                    Reason for decline: Reasons for decline of the Colorado squawfish
                    include the construction of dams and reservoirs, water withdrawals, and
                    competition and predation by non-native species.
                    Status: Endangered
References
Carter, J.G., R.A. Valdez, RJ. Ryel and V.A. Lamarra. 1985. Fisheries habitat dynamics in
       the upper Colorado River.  J. Freshwater Ecology. 3:249.

Tyus, H.M. 1991. Ecology and Management of Colorado Squawfish.  in Battle against
       extinction: Native fish  management in the American West. eds. W.L.  Minckley and
       I.E. Deacon 517 pp.

U.S. Fish and Wildlife Service.  1990.  Colorado squawfish recovery plan.  Denver, CO.
       56 p.
                                         36

-------
Appendix 1. Species profile - Razorback sucker (Xyrauchen texanus)

                    Historic range:  The historic range of the razorback sucker included the
                    upper and lower Colorado River basin and the Gila River.

                    Current range:  The razorback sucker is abundant only in Lake Mohave
                    and about 1000  razorbacks occur in the upper Green River.

                    Reasons for decline:  Causes for declines in razorback sucker
                    populations are  similar to those for Colorado squawfish; alterations of
                    the river environment including the construction of dams and potential
                    competition and predation by non-native species.
                    Status: Endangered
References
Carter, J.G., R.A. Valdez, RJ. Ryel and V.A. Lamarra. 1985. Fisheries habitat dynamics in
       the upper Colorado River.  J. Freshwater Ecology. 3:249.

Me Ada, C.W. and R.S. Wydoski.  1980. The razorback sucker, Xyrauchen texanus. in the
       upper Colorado River Basin,  1974-1976.  Technical papers of the USFWS/FWS,
       Washington, DC 15 p.

Minckley, W.L., P.C. Marsh, I.E. Brooks, I.E. Johnson and B.L. Jensen.  1991.
       Management toward recovery of the razorback sucker,  in Battle against extinction:
       Native fish  management in the American West. eds. W.L.  Minckley and J.E. Deacon
       517 pp.
                                          37

-------
APPENDIX 2. ARITHMETIC MEANS
             38

-------
Appendix 2.   Acute toxicity of carbaryl (mg/L) to 8 species of fish (2 surrogate and 6 listed)
              at 12, 24, and 96  h of exposure.  Toxicity values are the arithmetic means for
              the individual replicate LCSOs (Sappington, 1995).  The number of LCSOs
              used in the calculation is listed in parentheses.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bony tail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
6.87
(H)
3.62
(5)
7.40
(2)
4.65
(6)
10.9
(2)
7.00
(2)
>10.0
(NA)1
>10.0
(NA)1
24-h LC50
(n)
4.23
(16)
2.73
(5)
3.63
(3)
3.45
(6)
9.10
(2)
6.52
(5)
6.51
(3)
6.87
(5)
96-h LC50
(n)
2.00
(18)
1.53
(4)
1.59
(2)
2.18
(5)
5.20
(16)
3.36
(5)
3.23
(6)
4.39
(5)
IMA  —
 NA = not applicable
                                           39

-------
Appendix 2.  Acute toxicity of copper (mg/L) to 8 species of fish (2 surrogate and 6 listed)
             at 12, 24, and 96 h of exposure.  Toxicity values are the arithmetic means for
             the individual replicate LCSOs (Sappington, 1995).  The number of LC50s
             used in the calculation is listed in parentheses.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bony tail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
0.39
(12)
0.17
(4)
>0.03
(NA)1
0.32
(5)
1.53
(H)
0.31
(6)
>1.00
(NA)
>1.00
(NA)
24-h LC50
(n)
0.13
(14)
0.08
(2)
>0.03
(NA)
0.15
(6)
0.85
(12)
0.24
(6)
0.52
(4)
0.41
(3)
96-h LC50
(n)
0.08
(12)
0.068
(2)
>0.03
(NA)
0.072
(4)
0.49
(15)
0.22
(6)
0.43
(6)
0.28
(6)
IMA  —
 NA = not applicable
                                          40

-------
Appendix 2.  Acute toxicity of 4-nonylphenol (mg/L) to 8 species of fish (2 surrogate and 6
             listed) at 12, 24, and 96 h of exposure.  Toxicity values are the arithmetic
             means for the individual replicate LCSOs (Sappington, 1995).  The number of
             LC50s used in the calculation is listed in parentheses.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
0.36
(15)
0.31
(6)
0.37
(3)
0.27
(3)
0.36
(12)
0.57
(4)
0.44
(3)
0.31
(6)
24-h LC50
(n)
0.32
(13)
0.23
(6)
0.30
(3)
0.23
(3)
0.35
(17)
0.48
(5)
0.28
(3)
0.23
(5)
96-h LC50
(n)
0.25
(14)
0.17
(5)
0.156
(3)
0.17
(5)
0.32
(H)
0.29
(4)
0.24
(4)
0.20
(2)
                                          41

-------
Appendix 2.  Acute toxicity of pentachlorophenol (mg/L) to 8 species of fish (2 surrogate
             and 6 listed) at 12, 24, and 96 h of exposure.  Toxicity values are the
             arithmetic means for the individual replicate LCSOs (Sappington, 1995). The
             number of LCSOs used in the calculation is listed in parentheses.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
0.22
(12)
0.21
(5)
>0.01
(NA)1
0.26
(1)
0.35
(17)
0.44
(6)
0.23
(5)
0.52
(3)
24-h LC50
(n)
0.19
(13)
0.14
(5)
>0.01
(NA)
0.20
(3)
0.32
(17)
0.40
(3)
0.14
(2)
0.27
(3)
96-h LC50
(n)
0.15
(17)
0.104
(5)
>0.01
(NA)
0.17
(5)
0.27
(17)
0.23
(6)
0.13
(3)
0.27
(3)
'NA = not applicable
                                          42

-------
Appendix 2.   Acute toxicity of permethrin (/^g/L) to 8 species of fish (2 surrogate and 6
              listed) at 12, 24, and 96 h of exposure.  Toxicity values are the arithmetic
              means for the individual replicate LC50s (Sappington, 1995).  The number of
              LCSOs used in the calculation is listed in parentheses.
Species
rainbow trout
Apache trout
greenback cutthroat
Lahontan cutthroat
fathead minnow
bonytail chub
Colorado squawfish
razorback sucker
12-h LC50
(n)
5.4
(15)
3.9
(4)
>1.0
(NA)1
3.5
(6)
13.6
(11)
>25.0
(NA)
>25.0
(NA)
13.1
(3)
24-h LC50
(n)
3.9
(16)
2.5
(4)
>1.0
(NA)
2.0
(6)
9.7
(14)
22.2
(1)
8.2
(!)
7.0
(3)
96-h LC50
(n)
3.8
(18)
1.97
(4)
>1.0
(NA)
1.80
(7)
9.7
(17)
22.2
(1)
17.6
(3)
6.3
(5)
'NA = not applicable
                                           43

-------
APPENDIX 3. EXPOSURE pH
          44

-------
Appendix 3.  Exposure water pH for each test (species/Run/chemical) at 0 and 96 h.  The
pH range represents the lowest and highest pH measured for all exposure concentrations and
replicates within a test.
Species                    Chemical                  Oh                 96 h
rainbow trout
Run 1





Run 2





Run 3




-
Run 4





Run 5






control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

8.6-8.7
8.5-8.7
8.3-8.6
8.5-8.6
8.4-8.6
8.4-8.7
8.4-8.5
8.3-8.5
8.3-8.4
8.4-8.5
8.3-8.4
8.2-8.5
8.4-8.7
7.4-8.5
8.1-8.4
8.2-8.7
8.4-8.7
8.4-8.6
8.4-8.7
8.2-8.4
8.3-8.4
8.4-8.5
7.8-8.5
6.5-8.5
7.7
7.9-8.0
7.9-8.0
7.7-8.0
7.9
7.8-8.0

7.7-7.9
7.7-7.9
7.8-7.9
7.8-7.9
7.7-7.7
7.7-7.9
7.6-8.0
7.6-7.8
7.6-8.0
7.5-7.7
7.5-7.6
7.4-7.7
7.9-8.1
7.9-8.1

7.9-7.9
7.8-7.9
7.8-8.1
7.9-8.1
7.7-7.8

7.7-7^9
7.8-7.9
7.8-7.9
7.9~8.0
7.8-8.0
7.9
7.5-7.9
7.6-8.0
7.8-8.1
                                          45

-------
Appendix 3. continued
Species
Run 6

Apache trout
Run 1

Run 2

greenback
cutthroat trout
Run 1

Chemical
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
Oh
8.3-8.4
8.2-8.4
8.1-8.4
7.8-8.4
8.1-8.3
8.0-8.4

8.6-8.7
8.4-8.6
8.3-8.7
8.4-8.6
8,5-8.6
8.5-8.6
8.2-8.4
8.2-8.3
8.2-8.3
7.3-8.3
8.1-8.3
8.2

8.5-8.6
8.5-8.6
8.5-8.7
8.5-8.7
8.4-8.5
8.4-8.7
96 h
7.6-7.9
7.5-7.9
7.5-7.9
6.5-7.9
6.8-8.2
7.1-7.9

7.7-7.9
7.8-8.0
7.9-8.2
7.8-7.9
7.6-7.7
7.8-7.8
7.7-7.8
7.7-8.0
7.7-7.9
7.3-7.7
7.6
7.7

7.8-7.9
7.6-8.0
7.6-7.9
7.5-7.6
7.6-8.1
7.6-8.1
                                        46

-------
Appendix 3.  continued
Species
Lahontan
cutthroat trout
Run 1





Run 2





fathead minnow
Run 1





Run 2





Chemical


control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
Oh


8.4-8.5
8.3-8.4
8.3-8.4
8.4
8.4
8.4
8.2-8.4
8.0-8.4
8.2-8.3
8.2-8.3
8.2-8.3
8.1-8.3

8.4-8.5
8.4-8.5
8.2-8.5
8.3-8.4
8.4-8.5
8.4-8.5
8.4-8.5
8.4-8.5
7.8-8.4
8.4-8.5
8.3-8.5
7.9-8.5
96 h


7.6-8.0
7.9-8.2
7.8-8.0
7.9
7.7-7.9
7.9-8.0
7.7-7.8
7.5-7.9
7.7-7.8
7.6-7.7
7.6-7.9
7.6-7.9

7.9-8.2
8.0-8.1
8.0-8.3
7.7-8.1
7.8-7.9
7.7-8.2
7.9-8.2
' 7.9-8.1
8.0-8.2
7.8-8.1
7.8-8.0
7.8-8.1
                                       47

-------
Appendix 3. continued
Species
Run 3





Run 4





Run 5





Run 6





bonytail chub
Run 1





Chemical
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
Oh
8.5-8.6
8.2-8.6
8.1-8.4
8.3-8.6
8.0-8.6
7.5-8.5
8.3-8.4
8.1-8.4
8.0-8.3
8.1-8.3
7.9-8.3
7.9-8.3
9.5-9.8
9.2-9.5
8.8-9.5
8.2-9.2
8.8-9.3
8.4-9.7
7.1
6.9-7.2
6.7-7.1
5.5-6.9
6.6-6.9
6.5-6.9

8.5-8.6
7.9-8.5
8.0-8.5
8.0-8.6
8.3-8.6
8.3-8.6
96 h
7.8-8.0
7.5-8.0
7.9
7.6-7.8
7.6-7.9
7.7-7.8
7.7-8.0
7.6-7.9
7.6-8.2
7.5-7.9
7.4-7.6
7.4-7.9
7.2-7.4
7.4-7.5
7.5-7.7
7.5-7.6
7.3-7.6
7.3-7.6
6.7-7.9
7.7-7.8
7.8-7.9
7.8-8.2
7.8-7.9
7.7-8.0

7.8-8.0
7.7-8.0
7.6-7.8
7.7-7.9
7.5-7.9
7.6-7.9
                                         48

-------
Appendix 3. continued
Species
Run 2

Colorado squawfish
Run 1

Run 2

Razorback sucker
Run 1

Chemical
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin

control
carbaryl
copper
4-nonylphenol
pentachlorophenol
permethrin
Oh
9.5-9.8
8.4-9,5
8,9-9.4
8.3-9.5
9.3-9.5
8.7-9.3

8.4-8.5
8.2-8.5
8.3-8.4
8.4-8.5
8.4-8.5
8.4-8.5
7.1
6.2-7.2
6.7-7.1
6.4-6.9
6.9-7.1
6.7-7.1

8.4-8.5
8.3-8.5
8.3-8.4
8.3-8.5
8.3-8.6
8.3-8.5
96 h
7.2-7.4
7.5-7.6
7.5
7.4-7.6
7.4-7.5
7.4-7.6

7.9-8.2
8.1-8.3
8.0-8.6
8.1-8.2
8.0-8.1
8.0-8.3
6.7-7.9
7.7-8.1
7.8
7.8-8.0
7.7-7.8
"7.9-8.0

7.9-8.2
8.0-8.1
7.9-8.1
7.8-8. 1
7.9-8.1
7.6-8.1
                                         49

-------
Appendix 3.  continued
Species                    Chemical                  Oh                  96 h
Run 2                     control                    8.3-8.4              7.7-8.0
                           carbaryl                   8.0-8.3              7.7-8.0
                           copper                    7.9-8.2              7.9-8.0
                           4-nonylphenol              8.1-8.3              7.9-8.0
                           pentachlorophenol          8.2-8.5              7.6-8.0
                           permethrin                 8.2-8.3              7.6-8.2
                                           50

-------
APPENDIX 4. DISSOLVED OXYGEN CONCENTRATIONS
                    51

-------
Appendix 4.  Exposure replicates (rep) and concentrations (cone) for which dissolved oxygen
was below 40% saturation at 48 h of exposure or below 60% saturation at 96 h of exposure.
Dissolved oxygen concentration limits for the coldwater (12°C) studies are: 48 h - 6.4 mg/L;
96 h - 4.3 mg/L.  Dissolved oxygen concentration limits for the warmwater (22°C) studies
are: 48 h - 5.2 mg/L; 96 h - 3.5 mg/L.
Species
rainbow trout
Run 1



Run 2



















Chemical rep

carbaryl 1
2
pentachlorophenol 2
3
control 1
2
3
1
carbaryl i

2

3

copper 1

2

4-nonylphenol 1

2

3

cone

low
low
low
low
water
water
water
acetone
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
48h

4.2
3.8
4.2
4.0
5.9
4.9
4.5
6.1
3.1
3.8
3.2
4.9
3.6
4.8
4.8
5.0
4.4
4.8
4.5
5.0
6.1
5.0
5.5
4.1
96h

'



3.1

2.6

3.2
3.5


3.4







4.1
2.1

2.8
                                          52

-------
Appendix 4. continued
Species Chemical rep
Run 2 cont.
pentachlorophenol 1

2.

3

permethrin 1

2
3
Run 4 control 1
• 2
3
carbaryl 1

2 ''

3

4-nonylphenol 1

2'

3

pentachlorophenol 1

- ' 2

3

3
cone

low
medium
low
medium
low
medium
low
medium
low
medium
water
water
acetone
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
low
medium
low
48h

4.1
4.6
4.7
3.9
6.0
4.5
6.3
5.5
5.2

6.2
5.4
5.7
3.7
5.1
3.3
5.2
3.5.
4.7
4.7
4.8
4.9
4.2
5.4'
4.7
5.4
4.6
5.7
4.3
5.5
3.9

96h

2.8
3.4
3.2
3.3

3.3

3.3

3.1



3.5

3.0

2.6













3.2
                                        53

-------
Appendix 4. continued
Species
Run 4 cont.




Run 5


Apache trout
Run 1

Run 2
greenback cutthroat
Run 1













Chemical

permethrin



carbaryl



pentachlorophenol

pentachlorophenol
trout
carbaryl



copper
4-nonylphenol


pentachlorophenol





rep

1

2
3
1
2
3

1
2
1

2

3

3
1
2
3
1


2
3

cone

low
medium
low
low
medium
medium
medium

low
low
medium

low
medium
low
medium
medium
low
low
low
low
med
high
high
high
medium
48h

5,4
5.6
5.5
5.4








6.0
5,4
6.0
5.2
6.2
5.4
6.2
6.2
5.6
6.2
4.8
5.9
6.1
2.9
96h





3.2
3.4
3.9

3.4
3.6
4.2













3
2.4
                                         54

-------
Appendix 4. continued
Species
Run 1 cont.

Lahontan cutthroat
Run 1
Run 2

fathead minnow
Run 1
Run 2








Run 3
Chemical

permethrin
trout
pentachlorophenol
pentachlorophenol


4-nonylphenol
control
4-nonylphenol


pentachlorophenol



permethrin
4-nonylphenol
rep

3

1
1
2
3

3

1
2

3
1
2
3

1
2
3
2
3
cone

low
medium

medium
medium
medium
medium

medium
acetone
medium
low
medium
medium
low
medium
low
low
medium
high
high
high
medium
medium
48h

6.1
6.3

5.0
3.4
3.7
4.0


4.8
3.7
5.1
3.1
2.9
3.8
4.6
4.4
3.9
4.9
.4.7
5.2
4.0
96h



3.4



2.0

2.8
2.4
2.4
3.2
3.1
3.0

2.3
2.8
2.7
1.5
                    pentachlorophenol
medium
2.7
                                         55

-------
Appendix 4. continued
Species
Run 3 com.


Run 4









Run 5





Run 6







Chemical

permethrin

carbaryl

copper
4-nonylphenol

pentachlorophenol




carbaryl
pentachlorophenol



permethrin
control

4-nonylphenol
pentachlorophenol


permethrin

rep

1
2
1
3
3
1
3
1

2

3
1
1
2
3

2
1
2
3
1
2
3
2
3
cone

medium
medium
low
low
low
medium
medium
low
medium
low
medium
low
medium
medium
low
low
medium
medium
water
water
medium
low
low
low
high
high
48h



5.1
4.9
4.7
3.4
4.0
4.9
4.8
5.2
4.5
4.6

4.1
4.4

4.2

5.2
5.0
4.6
2.7
4.0
3.7
4.6
4.9
96h

1.7
1.2



2.5
3.2



3.0

2.8
3.2
3.4
3.0
2.0
3.5


3.5
2.9
3.3
2.8
2.8

                                        56

-------
Appendix 4. continued
Species
bonytail chub
Run 2


Colorado squawfish
Run 2
razorback sucker
Run 1
Run 2
Chemical

4-nonylphenol
pentacnlorophenol
permethrin

pentachlorophenol

permethrin
pentachlorophenol
rep

1
2
2'
2

1
3

1
2
3
cone

medium
medium
low
medium
medium

low
low

medium
high
medium
medium
48

4.9
4.1
5.1
4.9
4.7

5.0
5.0

5.0
5.2
4.7
96

3.0
4.2




1.6
1.8

                                        57

-------
APPENDIX 5. ACUTE TOXICITY DATA
              58

-------
   Appendix 5.  Calculated LG50, 95% confidence interval and slope for each chemical, species, Run and time period.  LCSO's and
   associated parameters were calculated using probit analysis unless otherwise noted.

                                                     CARBARYL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
RAINBOW TROUT 1
(0.67 ± 0.35)
RAINBOW TROUT 2
(1.25 ±0.57)
RAINBOW TROUT 3
(0.27 ±0.07)
RAINBOW TROUT 4
(1.09 ±0.38)
RAINBOW TROUT 5
(0.48 ± 0.08)
RAINBOW TROUT 6
(0.50 ±0.21)
HOURS
3
10.53
(8.8-15.7)
4.63
>10
8.2 c
(7.2-9.2)
>10
>6.0
>6.0
6
9.07
(7.4-12.6)
3.56
>10
7.11 °
(6.2-8.1)
>10
>6.0
>6.0
9
8.0
(6.8-10.1)
4.04
10.0
(8.3-14.2)
4.37
6.67
(5.5-8.7)
2.98
8.27
(7.1-10.4)
4.35
>6.0
>6.0
12
7.0
(6.1-8.3)
4.81
7.5
(6.5-8.7)
5.81
5.5
(4.7-6.7)
3.49
7.22
(6.3-8.6)
4.88
>6.0
>6.0
18
5.24
(4.6-6.0)
5.61
5.1
(4.5-5.9)
5.97
4.62
(4.0-5.4)
3.94
4.39
(3.8-5.1)
4.62
>6.0
>6.0
24
4.51
(3.2-6.6)
4.75
3.7
(3.2-4.4)
3.78
3.65
(3.2-4.2)
4.65
3.43
(3.0-3.9)
4.79
4.75°
(4.2-5.4)
4.41
(4.0-4.9)
11.10
48
2.S9
(2.5-3.3)
5.15
1.98
(1.7-2.3)
3.64
2.60
(2.3-3.0)
5.44
1.27
(1.1-1.4)
5.98
3.30
(2.9-3.8)
5.19
3.5
(3.2-3.8)
10.04
72
2.20
(1.9-2.5)
5.31
1.24
(1.0-1.5)
3.65
2.31
(2.1-2.6)
6.47
1.22
(1.1-1.4)
5.34
2.7
(2.4-3.0)
6.79
3.14
(2.8-3.5)
8.70
96
2.00
(1.8-2.3)
5.88
1.24
(1.0-1.5)
3.65
2.26
(2.0-2.5)
6.87
1.22
(1.1-1.4)
5.43
2.07
(1.9-2.3)
7.58
3.11 "
(2.6-3.6)
"-LC50 and 95 % CI calculated using moving average
b~LC50 calculated using nonlinear interpolation; CI calculated using binomial procedure
C-LC50 and 95 % CI calculated using Spearman-Karber
   a — LC50 and 95 % CI calculated using moving average
   b — LC50 calculated using nonlinear interpolation; CI calculated using binomial test (Stephan)
   c - LC50 calculated using Spearman-Karber

-------
Appendix 5, (cont.)
                                      CARBARYL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
APACHE TROUT 1
(0.85 ± 0.49)

APACHE TROUT 2
(0.38 ±0.1 8)

GREENBACK
CUTTHROAT TROUT
(0.31 ±0.17)
LAHONTAN
CUTTHROAT TROUT 1
(0.34 ±0.08)
LAHONTAN
CUTTHROAT TROUT 2
(0.57 ± 0.23)
HOURS
3
5.59
(4.9-6.4)
5.56
5.60
(4.2-10.0)
2.83
>10.0


5.20
(3.0-11.5)
4.01
4.60
(4.0-5.5)
5.11
6
4.96
(4.3-5.8)
4.15
3.75
(2.9-5.7)
2.42
10.00
(7.5-17.7)
2.25
4.91
(4.2-5.8)
3.93
4.6
(4.0-5.5)
5.11
9
4.64
(4.0-5.4)
4.20
2.99
(2.4-3.9)
3.02
9.89
(6.8-20.2)
1.66
4.59
(3.9-5.4)
3.73
4.35
(3.8-5.1)
5.26
12
4.16
(3.6-4.8)
4.11
2.60
(2.1-3.3)
3.22
8.50
(6.1-14.9)
1.80
4.48
(2.5-9.2)
3.94
4.28
(3.7-5.0)
5.21
18
3.55
(3.1-4.1)
4.65
2.12
(1.8-2.5)
4.99
5.08
(4.1-6.6)
2.39
4.26
(3.7-5.0)
4.08
4.0
(3.5-4.6)
6.13
24
3.12
(2.7-3.6)
4.46
2.00
(1.7-2.4)
4.71
3.59
(3.0-4.3)
3.05
3.56
(3.1-4.1)
4.93
3.62
(3.3-4.0)
8.48
48
2.02
(1.7-2.3)
4.20
1.66
(0.8-4.4)
4.77
2.30
(2.0-2.6)
4.61
2.53
(2.3-2.8)
7.08
3.06
(2.8-3.4)
10.56
72
1.74
(1.5-2.0)
4.88
1.45
(0.6-4.2)
5.20
2.19
'(1.9-2.5)
4.47
2.19
(1.96-2.4)
8.32
2.6 b
(2.2-3.6)

96
1.65
(1.5-1.9)
5.73
1.43 b
(1.3-2.2)

1.55
(1.4-1.8)
5.67
2.00
(1.8-2.2)
8.73
2.53"
(2.2-3.6)


-------
Appendix 5. (cont.)
                                      CARBARYL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
FATHEAD
MINNOW 1
(0.32 ±0.16)
FATHEAD
MINNOW 2
(0.56 ± 0. 19)
FATHEAD
MINNOW 3
(0.45 ± 0.35)
FATHEAD
MINNOW 4
(0.40 ± 0.21)
FATHEAD
MINNOW 5
(0.34 ± 0.24)
FATHEAD
MINNOW 6
(0.39 ±0.14)
HOURS
3
>10


>10


>16.7


>10


>10


>10


6
>10


>10


15.90
(14.1-18.8)
7.59
>10


>10


>10


9
>10


>10


12.0
(10.7-13.2)
8.91
>10


>10


>10


12
>10


>10


12.0
(10.7-13.2)
8.91
>10


>10


>10


18
>10


>10


12.0
(11.4-13.6)
14.0
>10


>10


>10


24
9.57 b
(6.0-10.0)

>10


10.0 b
(6.0-16.7)

>10


>10


>10


48
7.02 b
(6.0-10.0)

7.90
(7.1-8.8)
8.36
8.66 b
(6.0-10.0)

6.88 c
(6.1-7.7)
'
7.22b
(6.0-10.0)

>10


72
4.85
(4.4-5.4)
9.74
7.1
(6.5-7.8)
10.20
7.55"
(6.0-10.0)

4.69
(4.1-5.4)
4.53
5.86
(5.3-6.5)
9.69
7.36
(6.5-8.5)
6.01
96
4.38
(3.95-4.86)
8.69
5.86
(5.3-6.5)
9.69
7.43 b
(6.0-10.0)

3.94
(3.4-4.5)
4.69
5.24
(4.7-5.8)
9.52
5.72
(5.1-6.5)
6.45

-------
  Appendix 5. (cont.)
                                         CARBARYL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
BONYTAIL
CHUB 1
(0.29 ± 0.08)
BONYTAIL
CHUB 2
(0.52 ± 0.09)
COLORADO
SQUAWFISH 1
(0.32 ± 0.05)
COLORADO
SQUAWFISH 2
(0.34 ± 0.05)
RAZORBACK
SUCKER 1
(0.31 ±0.04)
RAZORBACK
SUCKER 2
(0.32 ± 0.07)
HOURS
3
>16.7
>10
>10
>10
>10
>10
6
0.90 b
(10-16.7)
>10
>10
>10
>10
>10
9
8.45
(7.5-9.6)
5.98
8.42 b
(6.0-10.0)
>10
>10
>10
>10
12
7.56"
(6.8-8.4)
8.31 b
(6.0-10.0)
>10
>10
8.88 c
(8.1-9.7)
>10
18
5.44 b
(3.6-6)
8.10
(7.4-8.8)
14.04
7.22 b
(6.0-10.0)
>10
7.19b
(6.0-10.0)
9.13
(8.3-10.2)
10.07
24
5.27 b
(3.6-6)
7.12 b
(6.0-10.0)
6.31 c
(5.7-7.0)
>10
5.83
(5.2-6.5)
7.49
7.64
(6.7-8.9)
6.03
48
5.21"
(4.6-5.9)
5.53 b
(3.6-6)
4.46 b
(3.6-6.0)
7.36 c
(6.9-7.8)
4.87
(4.4-5.3)
11.17
5.06
(4.6-5.6)
8.99
72
4.33 b
(3.6-6)
4.22"
(3.7-4.9)
2.91
(2.6-3.2)
9.36
5.42°
(4.9-6.0)
4.46 b
(3.6-6)
4.81
(4.3-5.3)
9.09
96
3.58
(3.3-4.0)
9.86
3.40 c
(3.0-3.7)
2.32
(2.1-2.6)
8.72
4.06 c
(3.7-4.4)
4.42 b
(3.6-6.0)
4.29
(4.0-4.8)
12.65
K)

-------
Appendix 4. (cont.)
                                           COPPER (mg/L)
SPECIES
AVG. WEIGHT
(grams)
RAINBOW TROUT 1
(0.67 ± 0.35)
RAINBOW TROUT 2
(1.25 ±0.57)
RAINBOW TROUT 3
(0.27 ± 0.07)
RAINBOW TROUT 4
(1.09 ±0.38)
RAINBOW TROUT 5
(0.48 ± 0.08)
RAINBOW TROUT 6
(0.50 ±0.21)
HOURS
3
->1,0
>0.6
>0.91
>0.91
>0.36
>0.36
6
>1.0
>0.6
>0.91
0.91
(0.73-1.35)
3.55
>0.36
>0.36
9
->1.0
>0.6
0.33
(0.17-0.86)
2.19
0.49 h
(0.33-0.55)
>0.36
>0.36
12
1.03
(0.8-1.6)
3.33
>0.6
0.15
(0.12-0.18)
2.69
0.42
(0.36-0.49)
3.77
>0.36
>0.36
18
0.33 -"
(0.29-0.39)
3.97
0.35
(0.31-0.39)
5.78
0.44
(0.01-0.07)
2.83
0.14
(0.12-0.17)
3.44
0.09
(0.08-0.10)
4.68
0.10
(0.09-0.12)
5.14
24
0.19
(0.17-0.21)
5.79
0.23
(0.20-0.27)
4.65
<0.07
0.07
(0.04-0.09)
2.57
0.07
(0.06-0.07)
6.11
0.07
(0.06-0.08)
7.80
48
0:13
(0.12-0.15)
6.96
0.17
(0.14-0.19)
3.69
<0.07
<0.07
0.06
(0.05-0.06)
9.65
0.06
(0.06-0.07)
9.71
72
0.11
(0.10-0.13)
5.62
0.13
(0.001-2.5)
2.41
<0.07
<0.07
0.05
(0.05-0.06)
9.92
0.06 b
(0.05-0.06)
9.28
96
"0.11
(0.09-0.12) •
6.02
0.10
(0.00003-0.32)
2.26
<0.07
<0.07
0.05 b
(0.05-0.09)
0.06 b
(0.05-0.09)

-------
Appendix 4. (cont.)
                                           COPPER (mg/L)
SPECIES
AVG. WEIGHT
(grams)
APACHE TROUT 1
(0.85 ± 0.49)

APACHE TROUT 2
(0.38 ±0.1 8)

GREENBACK
CUTTHROAT TROUT
(0.31 ±0.17)
LAHONTAN
CUTTHROAT TROUT 1
(0.34 ± 0.08)
LAHONTAN
CUTTHROAT TROUT 2
(0.57 ± 0.23)
HOURS
3
>1.0


>0.36


>0.03


>0.06


>0.36


6
0.76
(0.06-1.07)
2.64
>0.36


>0.03


>0.06


>0.36


9
0.34
(0.29-0140)
3.43
0.26
(0.21-0.39)
2.91
>0.03


>0.06


>0.36


12
0.21
(0.18-0.24)
3.41
0.16
(0.14-0.20)
4.10
>0.03


0.55
(0.47-0.68)
5.51
0.28
(0.25-0.32)
7.04
18
0.12
(0.01-0.13)
4.94
0.11
(0.09-0.12)
4.80
>0.03


0.23
(0.13-0.44)
3.62
0.15
(0.13-0.16)
8.01
24
0.10
(0.08-0.11)
4.67
0.08
(0.07-0.10)
5.95
>0.03


0.14
(0.12-0.16)
4.22
0.09
(0.08-0.10)
11.82
48
<0.08


0.07
(0.06-0.08)
6.08
>0.03


0.10
(0.06-0.16)
7.10
0.06 b
(0.05-0.08)

72
<0.08


0.07
(0.06-0.08)
6.08
>0.03


0.08 b
(0.08-0.13)

0.06 b
(0.05-0.08)

96
<0.08


0.07
(0.06-0.08)
6.08
>0.03


0.08
(0.07-0.09)
10.42
0.06 b
(0.05-0.08)


-------
Appendix 4. (cont.)
                                           COPPER (mg/L)
SPECIES
AVG. WEIGHT
(grams)
FATHEAD
MINNOW 1
(0.32 ±0.16)
FATHEAD
MINNOW 2
(0.56 ±0.19)
FATHEAD
MINNOW 3
(0.45 ± 0.35)
FATHEAD
MINNOW 4
(0.40 ±0.21)
FATHEAD
MINNOW 5
(0.34 ±0.24)
FATHEAD
MINNOW 6
(0.39 ±0.1 4)
HOURS
3
>1.0


>7.7


>4.6


>1.0


>1.0


>1.0


6
>1.0


4.43
(3.9-5.0)
5.99
3.58
(3.2-4.0)
8.86
>1.0


>1.0


>1.0


9
>1.0


3.22
(2.9-3.6)
6.57
1.83
(1.6-2.1)
3.68
>1.0


>1.0


>1.0


12
0.91
(0.8-1.1)
7.31
2.75
(2.5-3.1)
7.44
1.28
(1.1-1.5)
4.24
>1.0


>1.0


0.9
(0.8-1.2)
4.0
18
0.63
(0.6-0.7)
6.36
2.10
(1.1-4.9)
4.0
0.73
(0.6-0.8)
4.78
>1.0


>1.0


0.68°
(0.6-0.8)

24
0.58
(0,5-0.7)
6.83
1.70
(1.4-2.0)
3.43
0.53 a
(0.5-0.6)

>1.0


>1.0


0.55C
(0.5-0.6)

48
0.52
(0.5-0.6)
6.42
1.16
(0.9-1.4)
2.68
0.50 '
(0.4-0.6)

0.58 b
(0.4-0.6)

1.0
(0.8-1.7)
2.68
0.48
(0.4-0.6)
3.1
72
0.35
(0.2-0.6)
2.94
0.76
(0.4-1.0)
1.87
0.44
(0.4-0.5)
4.55
0.56
(0.3-3.7)
2.79
0.91
(0.7-1.5)
2.31
0.39
(0.3-0.5)
3.0
96
0.29
(0.1-0.6)
2.73
0.63
(0.3-0.9)
1.77
0.40
(0.3-0.5)
3.95
0.46
(0.4-0.6)
2.97
0.81
(0.6-1.3)
2.08
0.39
(0.3-0.5)
3.0

-------
Appendix 4. (cont.)
                                           COPPER (mg/L)
SPECIES
AVG. WEIGHT
(grams)
BONYTAIL CHUB 1
(0.29 ± 0.08)
BONYTAIL CHUB 2
(0.52 ± 0.09)
COLORADO
SQUAWFISH 1
(0.32 +. 0.05)
COLORADO
SQUAWFISH 2
(0.34 ± 0.05)
RAZORBACK
SUCKER 1
(0.31 +.0.04)
RAZORBACK
SUCKER 2
(0.32 _+ 0.07)
HOURS
3
1.33
(1.16-1.61)
5.03
>1.0
>1.0
>1.0
>1.0
>1.0
6
0.41
(0.37-0.45)
8.32
>1.0
>1.0
>1.0
>1.0
>1.0
9
0.29
(0.27-0.32)
10.59
0.57
(0.48-0.68)
3.84
>1.0
>1.0
>1.0
>1.0
12
0.25
(0.22-0.27)
9.49
0.35 b
(0.22-0.36)
>1.0
>1.0
>1.0
>1.0
18
0.23
(0.20-0.25)
7.57
0.28
(0.25-0.31)
8.81
0.69
(0.57-0.91)
2.91
>1.0
0.55
(0.47-0.64)
4.37
>1.0
24
0.22
(0.19-0.24)
8.31
0.27
(0.24-0.29)
10.05
0.46
(0.39-0.55)
3.53
0.89
(0.79-1.04)
6.9
0.39
(0.34-0.43)
7.06
>1.0
48
0.20
(0.18-0.22)
7.64
0.25 b
(0.22-0.36)
0.40
(0.35-0.46)
4.40
0.57 c
(0.51-0.64)
0.25
(0.22-0.29)
5.88
0.52
(0.46-0.60)
5.45
72
. 0.20
(0.18-0.22)
7.64
0.25 b
(0.22-0.36)
0.39
(0.33-0.45)
4.25
0.50
(0.23-4.7)
2.7
0.24
(0.21-0.27)
6.25
0.38
(0.33-0.43)
6.23
96
0.20
(0.18-0.22)
7.64
0.25 b
(0.22-0.36)
0.38
(0.33-0.44)
4.24
0.48
(0.22-4.2)
2.6
0.22
(0.19-0.25)
5.52
0.34"
(0.29-0.40)

-------
Appendix 4. (cont.)
                                  4-NONYLPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grains)
RAINBOW TROUT 1
(0.67 ± 0.35)
RAINBOW TROUT 2
(1.25 ±0.57)
RAINBOW TROUT 3
(0.27 4. 0.07)
RAINBOW TROUT 4
(1.09 ±0.38)
RAINBOW TROUT 5
(0.48 ± 0.08)
RAINBOW TROUT 6
(0.50 ±0.21)
HOURS
3
0.73
(0.67-0.80)
13.28
>0.6b
>0.36b
>0.6b
>0.6
>0.6
6
0.56
. (0.51-0.62)
10.09
0.54 b
(0.36-0.6)
0.39 h
(0.22-0.6)
0.59 b
(0.36-0.6)
0.45 b
(0.36-0.6)
0.46 b
(0.36-0.6)
9
0.43
(0.39-0.47)
12.65
0.45"
(0.36-0.6)
0.28
(0.26-0.31)
17.08
0.49 b
(0.36-0.6)
0.37 b
(0.36-0.6)
0.41 b
(0.36-0.6)
12
0.40 b
(0.22-0.6)
0.44 b
(0.36-0.6)
0.27 b
(0.22-0.36)
0.39 b
(0.22-0.6)
0.31 b
(0.22-0.6)
0.32 b
(0.22-0.36)
18
0.33 b
(0.22-0.36)
0.37
(0.34-0.41)
9.79
0.27 b
(0.22-0.36)
0.34
(0.31-0.37)
10.38
0.29
(0.26-0.31)
15.60
0.30"
(0.22-0.36)
24
0.31
(0.29- 0.34)
12.04
0.35
(0.32-0.39)
8.56
0.27 b
(0.22-0.36)
0.32
(0.29-0.35)
9.82
0.29
(0.26-0.31)
15.60
0.29 b
(0.22-0.36)
48
0.22
(0.20-0.24)
9.76
0.27
(0.24-0.30)
11.32
0.17b
(0.13-0.22)
0.27 b
(0.22-0.36)
0.18 b
(0.13-0.22)
0.22 b
(0.13-0.36)
72
0.19
(0.17-0.21)
9.10
0.26 b
(0.22-0.36)
0.14 b
(0.08-0.22)
0.27 b
(0.22-0.36)
0.16 b
(0.13-0.22)
0. 19 b
(0.13-0.22)
96
0.19
(0.17-0.21)
9.34
0.26 b
(0.22-0.36)
0.14b
(0.08-0.22)
0.27 b
(0.22-0.36)
0.16 b
(0.13-0.22)
0.18b
(0.13-0.22)

-------
  Appendix 4. (cont.)
                                     4-NONYLPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams) |
APACHE TROUT 1
(0.85 ± 0.49)
APACHE TROUT 2
(0.38 +.0.18)
GREENBACK
CUTTHROAT TROUT
(0.31 ±0.17)
LAHONTAN
CUTTHROAT TROUT 1
(0.34 +. 0.08)
LAHONTAN
CUTTHROAT TROUT 2
(0.57 ± 0.23)
HOURS
3
0.67 b
(0.6-1)
>0.6
0.55"
(0.36-0.6)
0.43"
(0.36-0.6)
0.56
(0.49-0.68)
8.60
6
0.44
(0.40-0.48)
13.28
0.41°
(0.38-0.45)
0.47
(0.42-0.51)
16.54
0.30 b
(0.22-0.36)
0.41 b
(0.36-0.6)
9
0.40 b
(0.36-0.6)
0.32 c
(0.29-0.35)
0.40 b
(0.22-0.6)
0.28 b
(0.22-0.36)
0.31 b
(0.22-0.36)
12
0.34 b
(0.22-0.36)
0.27 c
(0.25-0.30)
0.38 b
(0.22-0.6)
0.27 b
(0.22-0.36)
0.30 b
(0.22-0.36)
18
0.26
(0.24-0.29)
13.77
0.23 c
(0.21-0.25)
0.31
(0.28-0.34
10.16
0.24 b
(0.13-0.36)
0.27 b
(0.22-0.36)
24
0.25
(0.22-0.27)
11.18
0.22"
(0.19-0.25)
0.30
(0.27-0.33)
10.07
0.23 b
(0.13-0.36)
0.27 b
(0.22-0.36)
48
0.21
(0.19-0.23)
9.76
0.19"
(0.17-0.21)
0.19
(0.17-0.21)
11.33
0.18
(0.16-0.19)
11.47
0.26 b
(0.22-0.36)
72
0.19"
(0.17-0.21)
0.16"
(0.14-0.18)
0.17
(0.15-0.18)
12.19
0.16b
(0.13-0.22)
0.23 b
(0.22-0.36)
96
0.18"
(0.16-0.02)
0.16"
(0.13-0.22)
0.15
(0.14-0.17)
11.78
0.14h
(0.08-0.22)
0.22 b
(0.13-0.36)
o\
oo

-------
  Appendix 4. (cont.)
                                    4-NONYLPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
FATHEAD
MINNOW 1
(0.32 ±0.16)
FATHEAD
MINNOW 2
(0.56 ±0.19)
FATHEAD
MINNOW 3
(0.45 ± 0.35)
FATHEAD
MINNOW 4.
(0.40 ±0.21)
FATHEAD
MINNOW 5
(0.34 ± 0.24)
FATHEAD
MINNOW 6
(0.39 ±0.14)
HOURS
3
0.72
(0.66-0.79)
12.64
0.75 b
(0.6-1)
0.74b
(0.6-1)
0.82 b
(0.6-1)
0.71
(0.65-078)
10.20
0.87 b
(0.6-1)
6
0.42 b
(0.36-0.6)
0.48
(0.44-0.53)
11.87
0.55 b
(0.36-0.6)
0.61 b
(0.6-1)
0.40 b
(0.36-0.6)
0.58
(0.52-0.64)
9.8
9
0.34
(0.30-0.37)
8.92
0.43 b
(0,36-0.6)
0.48 b
(0.36-0.6)
0.44 b
(0.36-0.6)
0.31
(0.29-0.34)
12.04
0.48
(0.44-0.53)
11.8
12
0.30
(0.27-0.34)
7.64
0.42b
(0.36-0.6)
0.45 b
(0.36-0.6)
0.42 b
(0.36-0.6)
0.28
(0.26-0.31)
14.04
0.45
(0.42-0.50)
15.0
18
0.27
(0.24-0.30)
7.72
0.42 b
(0.36-0.6)
0.40 b
(0.36-0.6)
0.39 b
(0.36-0.6)
0.25 b
(0.22-0.36)
0.43 b
(0.36-0.6)
24
0.25
(0.22-0.27)
8.44
0.41 b
(0.36-0.6)
0.38 b
(0.22-0.6)
0.38"
(0.36-0.6)
0.21 "
(0.13-0.22)
0.39 b
(0.36-0.6)
48
0.24
(0.22-0.27)
8.09
0.40 b
(0.22-0.6)
0.34 b
(0.22-0.36)
0.36 b
(0.22-0.6)
0.17 b
(0.13-0.22)
0.31 b
(0.22-0.36)
72
0.23
(0.21-0.25)
9.96
0.36 b
(0.22-0.6)
0.31 b
(0.22-0.36)
0.35 b
(0.22-0.36)
0.17 b
(0.13-0.22)
' 0.30 b
(0.22-0.36)
96
0.21 b
(0.13-0.22)
0.36 b
(0.22-0.6)
0.31 b
(0.22-0.36):
0.33 b
(0.22-0.36)
0,17 b
(0.13-0.22)
0.29 b
(0.22-0.36)
Os

-------
Appendix 4. (cont.)
                                  4-NONYLPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
BONYTAIL
CHUB 1
(0.29 ± 0.08)
BONYTAIL
CHUB 2
(0.52 ± 0.09)
COLORADO
SQUAWFISH 1
(0.32 ± 0.05)
COLORADO
SQUAWFISH 2
(0.34 +. 0.05)
RAZORBACK
SUCKER 1
(0.31 ± 0.04)
RAZORBACK
SUCKER 2
(0.32 ± 0.07)
HOURS
3
0.63 b
(0.36-1)
>1.0
0.77 b
(0.6-1)
0.80 b
(0.6-1)
0.73 b
- (0.6-1)
>1.0
6
0.46 b
(0.36-0.6)
.78"
(0.6-1)
0.63 b
(0.36-1)
0.63 b
(0.6-1)
0.41
(0.37-0.46)
8.24
0.63
(0.57-0.70)
4.18
9
0.45 b
(0.36-0.6)
0.74 b
(0.6-1)
0.48 b
(0.36-0.6)
0.47 b
(0.36-0.6)
0.29
(0.26-0.33)
7.11
0.44
(0.40-0.49)
3.45
12
0.44 b
(0.36-0.6)
0.72 b
(0.6-1)
0.44
(0.40-0.48)
13.28
0.45 b
(0.36-0.6)
0.27
(0.24-0.30)
7.41
0.31
(0.28-0.34)
3.89
18
0.38 b
(0.22-0.6)
0.68"
(0.6-1)
0.31 b
(0.22-0.36)
0.31 b
(0.22-0.36)
0.23
(0.21-0.26)
7.80
0.25 b
(0.22-0.36)
24
0.38 b
(0.22-0.6)
0.62 b
(0.6-1)
0.28
(0.26-0.31)
14.04
0.28 b
(0.22-0.36)
0.21
(0.19-0.23)
9.65
0.23 b
(0.22-0.36)
48
0.34
(0.31-0.38)
10.19
0.38 h
(0.36-0.6)
0.26 h
(0.22-0.36)
0.27 h
(0.22-0.36)
0.19"
(0.13-0.22)
0.22 h
(0.13-0.22)
72
0.27 b
(0.22-0.36)
0.35 b
(0.22-0.36)
0.25 b
(0.22-0.36)
0.27 b
(0.22-0.36)
0.18 b
(0.13-0.22)
0.20 b
(0.13-0.22)
96
0.27 b
(0.22-0.36)
0.31 b
(0.26-0.37)
0.24 b
(0.13-0.36)
0.27 b
(0.22-0.36)
0.16 b
(0.13-0.22)
0.19b
(0.13-0.22)

-------
Appendix 4. (cont.)
                              PENTACHLOROPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams) J
RAINBOW TROUT 1
(0.67 ± 0.35)
RAINBOW TROUT 2
(1.25 ± 0.57)
RAINBOW TROUT 3
(0.27 ± 0.07)
RAINBOW TROUT 4
(1.09 ±0.38)
RAINBOW TROUT 5
(0.48 ± 0.08)
RAINBOW TROUT 6
(0.50 ±0.21)
HOURS
3
0.43 b
(0.36-0.06)
0.27
(0.25-0.30)
15.60
>0.60
>0.60
>0.36»
>0.36
6
0.30 b
(0.22-0.36)
0.21 b
(0.13-0.22)
0.29 b
(0.22-0.36)
0.31 b
(0.22-0.36)
>0.36
>0.36
9
0.21
(0.19-0.23)
9.92
0.18 b
(0.13-0.22)
0.26 °
(0.24-0.28)
0.27
(0.25-0.29)
14.57
0.29
(0.27-0.32)
14.57
>0.36
12
0.18
(0.17-0.20)
12.10
0.17b
(0.13-0.22)
0.23
(0.20-0.25)
8.16
0.21 b
(0.13-0.22)
0.26 b
(0.22-0.36)
0.29 b
(0.22-0.36)
18
0.20
(0.18-0.22)
10.35
0.15b
(0.13-0.22)
0.18b
(0.016-0.20)
0.17b
(0.13-0.22)
0.25 b
(0.22-0.36)
0.27 b-
(0.22-0.36)
24
0.13 b
(0.08-0.22)
0.13b
(0.13-0.22)
0.16"
(0.14-0.18)
0.17b
(0.13-0.22)
0.22 b
(0.13-0.36)
0.25 b
(0.22-0.36)
48
0.11
(0.10-0.12)
11.63
0.12
(0.11-0.13)
11.04
0.15"
(0.14-0.17)
0.15b
(0.13-0.22)
0.19b
(0.13-0.22)
0.19b
(0.13-0.22)
72
0.11
(0.95-0.12)
8.61
0.12
(0.11-0.13)
11.04
0.14
(0.13-0.16)
6.6
0.15b
(0.13-0.22)
0.19 b
(0.13-0.22)
0.19b
(0.13-0.22)
96
0.15"
(0.13-0.22)
0.12b
(0.08-0.13)
0.15b
(0.13-0.22)
0.15b
(0.13-0.22)
0.18 b
(0.13-0.22)
0.19b
(0.13-0.22)

-------
  Appendix 4. (cont.)
                                PENTACHLOROPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
APACHE TROUT 1
(0.85 ±0.49)
APACHE TROUT 2
(0.38 ±0.18)
GREENBACK
CUTTHROAT TROUT
(0.31 ±0.1 7)
LAHONTAN
CUTTHROAT TROUT 1
(0.34 ±0.08)
LAHONTAN
CUTTHROAT TROUT 2
(0.57 ± 0.23)
HOURS
3
0.35 b
(0.22-0.36)
>0.36
>0.01
>0.36
>0.36
6
0.28
(0.25-0.30)
13.02
0.32 b
(0.22-0.36)
>0.01
0.30 b
(0.22-0.36)
>0.36
9
0.21
(0.19-0.23)
9.80
0.27 b
(0.25-0.31)
13.69
>0.01
0.28 b
(0.22-0.36)
>0.36
12
0.19
(0.17-0.20)
11.77
0.23
(0.20-0.26)
8.81
>0.01
0.27 b
(0.22-0.36)
>0.36
18
0.16b
(0.13-0.22)
0.17'
(0.15-0.19)
11.25
>0.01
0.23 b
(0.22-0.36)
0.29 b
(0.22-0.36)
24
0.30 b
(0.08-0.22)
0.15
(0.13-0.17)
8.07
>0.01
0.19b
(0.13-0.22)
0.28 b
(0.22-0.36)
48
0.11
(0.10-0.12)
12.19
0.12
(0.10-0.13)
6.55
>0.01
0.18
(0.17-0.20)
12.29
0.18
(0.17-0.20)
12.29
72
0.11
(0.10-0.12)
9.70
0.10
(0.09-0.12)
5.67
>0.01
0.18"
(0.16-0.20)
0.18
(0.17-0.20)
12.29
96
0.11
(0.10-0.12)
8.97
0.10
(0.08-0.12)
5.05
>0.01
0.16
(0.08-0.36)
6.21
0.18 b
(0.13-0.22)
to

-------
Appendix 4. (cont.)
                              PENTACHLOROPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
FATHEAD
MINNOW 1
(0,32 ±0.16)
FATHEAD
MINNOW 2
(0.56 ±0.19)
FATHEAD
MINNOW 3
(0.45 ± 0.35)
FATHEAD
MINNOW 4
(0.40 ±0.21)
FATHEAD
MINNOW 5
(0.34 ± 0.24)
FATHEAD
MINNOW 6
(0.39 ±0.14)
HOURS
3
0.74 b
(0.6-1)
0.84
(0.76-0.91)
12.63
0.72
(0.66-0.79)
12.64
>1.0
0.98 b
(0.6-1)
0,34 b
(0.22-0.36)
6
0.52
(47-56)
11.73
0.51 b
(0.36-0.6)
0.50
(0.46-0.55)
12.64
0.64 b
(0.6-1)
0.68 b
(0.6-1)
0.21 k
(0.13-0.22)
9
0.38
(0.34-0.42)
6.94
0.38 b
(0.36-0.6)
0.40 b
(0.36-0.6)
0.47
(0.42-0.51)
16,54
0.55 b
(0.36-0.6)
0.16
(0.15-0.18)
9.99
12
0.34
(0.31-0.39)
7.06
0.32 k
(0.22-0.36
0.34 b
(0.22-0.36)
0.44
(0.41-0.49)
14.05
0.54 b
(0.36-0.6)
0.15
(0.13-0.16)
10.6
18
0.31
(0.27-0.35)
6.39
0.30 b
(0.22-0.36)
0.34 b
(0.22-0.36)
0.36
(0.33-0.40)
9.74
0.54 b
(0.36-0.6)
0.14
(0.13-0.16)
10.4
24
0.29
(0.26-0.33)
5.80
0.28
(0.26-0.31)
12.0
• 0.34 b
(0.22-0.36)
0.33"
(0.30-0.36)
0.53
(0.48-0.59)
9.26
0.14
(0.13-0.16)
10.4
48
0.29
(0.25-0.33)
5.91
0.26
(0.23-0.29)
9.24
0.33
(0.30-0.36)
10.89
0.31
(0.28-0.34)
8.72
0.50
(0.45-0.55)
7.69
0.14
(0.13-0.16)
10.4
72
0.29
(0.25-0,33)
5.91
0.23
(0.21-0.26)
7.75'
0.29
(0.13-0.67)
5.68
0.3P
(0.27-0.34)
0.46
(0.41-0.51)
7.27
0.14
(0.13-0.16)
10.4
96
0.27
(0.24-0.31)
5.89
0.22
(0.20-0.24)
8.39
•0.23
(0.11-0.49)
4.15
0.28
(0.16-0.48)
7.94
0.44a
(0.39-0.50)
0.14
(0.13-0.16)
10.2

-------
Appendix 4. (cont.)
                              PENTACHLOROPHENOL (mg/L)
SPECIES
AVG. WEIGHT
(grams)
BONYTAIL CHUB 1
(0.29 ± 0.08)
BONYTAIL CHUB 2
(0.52 ± 0.09)
COLORADO
SQUAWFISH 1
(0.32 ± 0.05)
COLORADO
SQUAWFISH 2
(0.34 +. 0.05)
RAZORBACK
SUCKER 1
(0.31 ±0.04)
RAZORBACK
SUCKER 2
(0.32 +. 0.07)
HOURS
3
0.76 b
(0.6-1)
>1.0
0.78 b
(0.6-1)
0.28 b
(0.22-0.36)
0.79
(0.72-0.86)
16.75
>1.0
6
0.51 b
(0.36-0.6)
0.78
(0.71-0.85)
16.54
0.51 b
(0.36-0.6)
0.17 b
(0.13-0.22)
0.56 b
(0.36-0.6)
0.84b
(0.6-1)
9
0.44 b
(0.36-0.6)
0.66 b
(0.6-1)
0.47 b
(0.36-0.6)
0.17 b
(0. 13-0.22)
0.45 b
(0.36-0.6)
0.73 b
(0.6-1)
12
0.33 b
(0.22-0.36)
0.54 b
(0.36-0.6)
0.40 b
(0.36-0.6)
0.13 b
(0.08-0.13)
0.45 b
(0.36-0.6)
0.63 b
(0.6-1)
18
0.28 b
(0.22-0.36)
0.43 b
(0.36-0.6)
0.28 b
(0.22-0.36)
0.10"
(0.08-0.13)
0.34
(0.31-0.38)
8.71
0.47 b
(0.36-0.6)
24
0.28 b
(0.22-0.36)
0.40 b
(0.36-0.6)
0.27 b
(0.22-0.36)
0.10b
(0.08-0.13)
0.27 b
(0.22-0.36)
0.47 b
(0.36-0.6)
48
0.24 b
(0.22-0.36)
0.28
(0.25-0.30)
15.60
0.23 b
(0.22-0.36)
0.10b
(0.08-0.13)
0.27 b
(0.22-0.36)
0.32"
(0.22-0.36)
72
0.22 b
(0.13-0.22)
0.27 b
(0.22-0.36)
0.18 b
(0.13-0.22)
0.10b
(0.08-0.13)
0.27 b
(0.22-0.36)
0.29 b
(0.22-0.36)
96
0.20 b
(0.13-0.22)
0.26 b
(0.22-0.36)
0.18 b
(0.13-0.22)
0.10b
(0.08-0.13)
0.27' b
(0.22-0.36)
0.28 b
(0.22-0.36)

-------
Appendix 4. (cont.)
                                        PERMETHRIN Qig/L)
SPECIES
AVG. WEIGHT
(grams)
RAINBOW TROUT 1
(0.67 ± 0.35)

RAINBOW TROUT 2
(1.25 ±0.57)
RAINBOW TROUT 3
(0.27 +. 0.07)

RAINBOW TROUT 4
(1.09 ±0.38)

RAINBOW TROUT 5
(0.48 ± 0.08)

RAINBOW TROUT 6
(0.50 ±0.21)

HOURS
3
>10


>10

>10


>10


>10


>10


6
>10


>10

7.62°
(6.9-8.5)

>10


>10


>10


9
7.16
(6.6-7.9)
12.64
9.57 b
(6-10)
4.42
(4.0-4.9)
8.73
8.47
(7.6-9.5)
8.66
6.94
(6.35-7.64)
11.62
9.72
(8.6-11:8)
7.26
12
5.96
(5.4-6.6)
9.58
8.31 b
(6-10)
3.39b
(2.2-3.6)

6.37
(5.7-7.1)
8.55
5.13b
(3.6-6)

- 6.62 b
(6-10)

18
2.97
(2.7-3.2)
11.58
6.39 b
(6-10)
2.64
(2.3-3.0)
6.42
5.00
(4.6-5.5)
12.64
3.39 b
(2.2-3.6)

4.16b
(3.6-6)

24
4.20
(2.4-7.6)
5.82
6.00 b
(3.6-10)
2.18
(1.9-2.4)
6.92
4.60
(4.2-5.0)
12.55
3.08
(2.8-3.4)
12.54
3.74
(3.4-4.1)
9.79
48
4.00
(3.5-4.5)
5.95
6.00 b
(3.6-10)
1.65
(1.5-1.9)
6.30
4.47
(4.1-4.9)
11.80
2.86
(2.6-3.2)
9.85
3.58
(3.2-3.9)
9.86
72
3.87
(3.4-4.4)
6.20
6.00 b
(3.6-10)
1.65
(1.5-1.9)
6.30
4.40
(4.0-4.8)
11.18
2.67
(2.4-2.9)
11.57
3.58
(3.2-3.9) .
9.86
96
3.90
(3.4-4.4)
6.20
4.80 a
(4.3-5.3)
1.65
(1.5-1.9)
6.30
4.40
(4.0-4.8)
11.18
2.67
(2.4-2.9)
11.57
3.60
(3.2-3.9)
9.86

-------
Appendix 4. (cont.)
                                       PERMETHRIN (pg/L)
SPECIES
AVG. WEIGHT
(grams)
APACHE TROUT 1
(0.85 ± 0.49)
APACHE TROUT 2
(0.38 ±0.18)
GREENBACK
CUTTHROAT TROUT
(0.31 ±0.17)
LAHONTAN
CUTTHROAT TROUT 1
(0.34 ±0.08)
LAHONTAN
CUTTHROAT TROUT 2
(0.57 ± 0.23)
HOURS
3
>10

>10

>1.0


>10

>10

6
8.07
(7.3-8.8)
11.68
>10

>1.0


4.42
(4.0-4.9)
8.73
>10

9
5.07
(4.6-5.6)
10.21
5.90
(4.5-8.2)
4.42
>1.0


2.81
(2.6-3.1)
14.04
5.71
(5.3-6.2)
10.16
12
4.07
(3.7-4.5)
9.0
3.70
(2.8-4.8)
4.44
>1.0


2.38
(2.2-2.6)
10.55
4.65
(4.3-5.0)
14.83
18
2.97
(2.7-3.2)
11.58
2.06 b
(1.3-2.2)
>1.0


1.75
(1.6-1.9)
8.83
3.06 b
(2.2-3.6)
24
2.63
(2.4-2.9)
9.61
1.96
(1.6-2.4)
7.99
>1.0


1.42
(1.3-1.6)
8.62
2.61 b
(2.2-3.6)
48
2.21
(2.0-2.4)
9.56
1.61
(1.3-2.0)
7.23
>1.0


1.16
(1.0-1.3)
7.37
2.23
(2.0-2.4)
10.05
72
2.17
(2.0-2.4)
9.59
1.39
(1.1-1.7)
7.66
>1.0


1.14
(1.0-1.3)
7.48
2.20
(2.0-2.4)
10.05
96
2.2
(1.96-2.4)
9.59
1.33
(1.1-1.7)
7.46
>1.0


1.14
(1.0-1.3)
7.48
2.20
(2.0-2.4)
10.05

-------
Appendix 4. (cont.)
                                        PERMETHRIN (pg/L)
SPECIES
AVG. WEIGHT
(grams)
FATHEAD
MINNOW 1
(0.32 ±0.16)
FATHEAD
MINNOW 2
(0.56 ±0.19)
FATHEAD
MINNOW 3
(0.45 ± 0.35)
FATHEAD
MINNOW 4
(0.40 ± 0.21)
FATHEAD
MINNOW 5
(0.34 ± 0.24)
FATHEAD
MINNOW 6
(0.39 ±0.1 4)
HOURS
3
>25


>10


>25


>25


>25


>25


6
>25


>10


>25


>25


>25


>25


9
13.91"
(9-15)

>10


17.49
(15.0-21.1)
4.3
>25


12.53
(10.5-15.3)
3.14
>25


12
13.43"
(11.5-16.0)

>10


13.64
(11.9-15.7)
5.03
17.32
(14.6-21.8)
3.58
10.25
(8.7-12.3)
3.23
>25


18
10.02
(8.5-11.9)
3.55
9.94
(8.7-12.8)
6.13
11.36
(9.9-13.2)
4.48
10.04
(5.6-18.8)
4.42
9.75
(8.2-11.8)
3.01
>25


24
9.24
(7.9-11.0)
3.41
9.48
(8.2-12.1)
5.51
11.11
(9.7-12.8)
4.77
9.69
(5.65-16.8)
4.72
9.26
(7.7-11.3)
2.72
>25


48
8.55
(7.4-10.0)
3.89
9.21
(8.0-11.4)
5.80
10.34
(9.1-11.7)
5.57
8.74"
(7.6-10.0)

8.93
(7.4-10.9)
2.74
16.8
(14.2-20.9)
3.6
72
8.09
(7.0-9.4)
4.31
9.21
(8.0-11.4)
5.80
10.0
(8.7-11.4)
5.00
8.74"
(7.6-10.0)

8.27
(6.9-9.9)
2.99
15.9
(13.6-19.3)
3.9
96
8.09
(7.0-9.4)
4.31
8.97
(7.9-10.9)
6.10
10.0
(9.1-11.7)
5.57
8.96"
(7.7-10.7)

6.68
(5.8-7.7)
4.50
15.7
(13.4-18.9)
3.9

-------
    Appendix 4. (cont.)
                                            PERMETHRIN (/ig/L)
SPECIES
AVG. WEIGHT
(grams)
BONYTAIL CHUB 1
(0.29 ± 0.08)
BONYTAIL CHUB 2
(0.52 +. 0.09)
COLORADO
SQUAWFISH 1
(0.32 +. 0.05)
COLORADO
SQUAWFISH 2
(0.34 +. 0.05)
RAZORBACK
SUCKER 1
(0.31 +.0.04)
RAZORBACK
SUCKER 2
(0.32 _+ 0.07)
HOURS
3
>25
>25
>10
>25
>25
>25
6
>25
>25
>io
>25
>25
>25
9
>25
>25
>10
>25
>25
>25
12
>25
>25
>10
>25
>25
13.05
(11.2-15.4)
4.05
18
>25
>25
>10
>25
>25
9.04
(3.6-22.9)
3.34
24
>25
>25
>10
>25
>25
8.87
(3.6-21.5)
3.26
48
>25
>25
>10
>25
17.43
(13.1-28.4)
1.86
8.23 .
(1.8-26.4)
3.03
72
>25
>25
>10
>25
4.97 b
(3.2-5.4)
7.95
(3.0-17.3)
3.00
96
>25
>25
>10
24.4 b
(15-25)
4.6 b
(3.2-5.4)
7.70
(3.1-15.9)
3.49
 oo
§

-------