BOSTON PUBLIC LIBRARY
3 9999 06317 776 8
7/
LETHAL DIETARY TOXICITIES
OF ENVIRONMENTAL POLLUTANTS
TO BIRDS
iTORY
UNITED STATES DEPARTMENT OF THE INTERIOR
FISH AND WILDLIFE SERVICE
Special Scientific Report— Wildlife No. 191
Library of Congress Cataloging in Publication Data
Main entry under title:
Lethal dietary toxicities of environmental pollutants to birds.
(Special scientific report-wildlife; no. 191)
Includes bibliographical references.
Supt. of Docs, no.: I 49.15/3:191
1. Pesticides and wildlife. 2. Pesticides— Toxicology. 3. Pollution-
Toxicology. 4. Game and game-birds— Diseases. I. Hill, Elwood F.
II. Series: United States. Fish and Wildlife Service. Special scientific
report-wildlife; no. 191. SK361.A256 no. 191 [QH545.P4] 639'.97'90973s
[639'.97'82] 75-619346
UNITED STATES DEPARTMENT OF THE INTERIOR
FISH AND WILDLIFE SERVICE
LETHAL DIETARY TOXICITIES
OF ENVIRONMENTAL POLLUTANTS
TO BIRDS
By
Elwood F. Hill, Robert G. Heath
James W. Spann, and Joseph D. Williams
Patuxent Wildlife Research Center
Laurel, Maryland 20811
^8 mst^
U.S. Fish and Wildlife Service
Special Scientific Report — Wildlife No.
Washington, D.C. 1975
191
CONTENTS
ABSTRACT iv
INTRODUCTION 1
PROCEDURES 1
RESULTS AND DISCUSSION 3
Comparative Toxicity in Relation to Chemical Class 3
Comparative Toxicity in Relation to Species 4
CONCLUSIONS 5
ACKNOWLEDGEMENTS 6
REFERENCES 7
Table 1 8
Table 2 37
Table 3 38
Table 4 40
Table 5 42
APPENDIX 43
in
ABSTRACT
This report is a compilation and analysis of the results of nearly
10 years of testing the lethal dietary toxicities of pesticidal and
industrial chemicals to young bobwhites (Colinus virginianus) , Japanese
quail (Coturnix c^ japonica) , ring-necked pheasants (Phasianus
colchicus) , and mallards (Anas platyrhynchos) .
A total of 131 compounds were tested. Toxicities are expressed
as median lethal dietary concentrations (LC50) and are based on 5 days
of dietary exposure to the test compound followed by 3 days of untreated
feed. From these data statistical comparisons between toxicities are
possible for a given species.
Certain classes of pesticides — organochlorine compounds, organo-
phosphates and organometallic compounds — contained most of the
compounds judged "highly toxic". The most frequent order of species
response was bobwhite > Japanese quail > ring-necked pheasant >
mallard. This order correlates with their body sizes at the ages
tested.
IV
INTRODUCTION
This report is a compilation of the results of nearly 10 years of
testing the subacute toxicities of pesticides and industrial chemicals
to young bobwhites (Colinus virginianus) , Japanese quail (Coturnix c_.
japonica) , ring-necked pheasants (Phasianus colchicus) and mallards
(Anas platyrhynchos) . It supersedes our earlier publication (Heath
et al. 1972). A total of 131 compounds were tested, including 30
organochlorine compounds, 39 organophosphates, 17 carboxylates, 15
carbamates, 12 metallic compounds, 5 organonitrogen compounds, 4
organosulfates, 4 ureas, 3 ketones, and 2 nitrophenols .
Our objectives were two-fold: to provide a readily referable
source of subacute toxicity data for the species and chemicals we
tested, and to compare the responses of different species to different
classes of chemicals.
A detailed exposition of mathematical procedures and a list of
chemical and common names (with often-used synonyms) are included in
appendices.
PROCEDURES
Subacute toxicity tests were designed to measure a median lethal
dietary concentration (LC50) of chemical to young birds during an
8-day test, including 5 days of treated diet followed by 3 days of
untreated diet. Five or six geometrically arranged concentrations of
toxicant were used per test at levels expected to kill from 10 to 90%
of the test population. An equal number of positive and negative controls
accompanied each test. Using shared controls, several compounds could be
tested simultaneously. A completely randomized experimental design was
used.
Feed treated with dieldrin dissolved in corn oil served as the
positive control (standard) and feed treated with corn oil, the diluent
used for most compounds tested, was the negative control.
Each test group (one test group per toxic concentration) consisted
of 10 birds. Ages of test birds were 14 days for quail and 10 days for
pheasants and mallards. In the 1973 tests, mallards were 5 days old.
Testing commenced at midday. Mortality and feed consumption were
recorded at 24-hour intervals thereafter. Fresh feed was added to all
pens daily. After the 5th day, all feed, including that of controls,
was replaced by untreated feed.
Occasional deviations from the basic procedure were necessary
because of shortages of facilities or birds. As few as five birds per
test group and four toxic concentrations per test were sometimes used.
Before standardization in 1970, age of birds sometimes varied as much
as 1 week between tests.
All test birds were incubator-hatched progeny of randomly outbred
Patuxent colonies. Bobwhite and pheasant colonies originated from the
Pennsylvania Game Commission's game farm stock; Japanese quail from
Auburn University; and mallards from wild stock. Parent colonies were
outbred to maximize individual variation and to more closely approximate
characteristics of wild populations.
Gallinaceous birds were tested in six-tiered brood units with tiers
divided into four pens measuring 35 x 100 x 24 cm. Floors and external
walls were of wire mesh; ceilings and common walls were galvanized sheet-
ing. Tiers were equipped with thermostatically controlled heat and
fluorescent lighting. Mallards were tested in wooden, walk- in pens on
concrete slabs. The pens measured 1.5 x 3.0 x 2.1 m. The upper half of
the lee wall of the pen was screened. Heat (infra-red), straw litter, and
running water were provided.
Test diets were prepared by blending a toxicant-carrier solution
into commercial starter mash in the ratio of 2:98, by weight. Corn oil
was the usual carrier, although propylene glycol was substituted when
compounds were insoluble in oil. Chemicals were dissolved in the carrier,
over heat when necessary. Some chemicals were first dissolved in minimal
quantities of acetone. If extremely large quantities of a compound were
required, or if the compound had a talc base, it was mixed directly into
the feed and appropriate amounts of the carrier were added to the mixture
as a supplement. Unstable compounds were mixed immediately before the
test. Control diets contained corn oil in the ratio of 2:98, by weight.
The LC50's and associated statistics were derived by methods of
probit analysis described by Finney (1952) and were programmed for com-
puter by the system of Daum and Killcreas (1966). The 50% response level
was chosen because it can be estimated more precisely than extreme
percentage levels (Finney 1952; see Appendix 1 for statistical details).
Positive and negative control data were included in the probit analysis
with every set of compounds tested simultaneously. Compounds with LC50's
exceeding 5000 ppm in preliminary range finding experiments were not
tested further. Estimates for preliminary data were made graphically by
the method of Litchfield and Wilcoxon (1949) and are presented as approxi-
mate values that are considered to be provisional.
In Table 1, the toxicity of each compound to each species is compared
,with the toxicity of the dieldrin standard to the same species as deter-
mined by concurrent tests. The resultant value (relative toxicity of
dieldrin or RTD) is a direct ratio of the LC50's of the two compounds and
represents the difference in toxicity between those two compounds under a
single condition. Therefore, toxicities of different chemicals in relation
to each other can be estimated statistically by using the RTD's as
described in Appendix 2. Lethal concentrations in addition to the LC50's
can also be estimated from data in Table 1 as described in Appendix 2.
RESULTS AND DISCUSSION
Five-day subacute dietary toxicities of 131 compounds were deter-
mined for young bobwhites, Japanese quail, ring-necked pheasants and
mallards. In Table 1, results are arranged alphabetically by common
name of the test compound. Chemical names, purity, chemical classes,
and principal uses are shown in Appendix 3.
Comparative Toxicity in Relation to Chemical Class
Although interest in toxicity data tends to center on specific
comparisons between chemicals, broader generalization is also useful
in understanding the toxicity process, especially if judgments of
relative toxicity are desired. Several rating systems have been
developed for this purpose (Hodge and Sterner 1949, Radeleff 1964, and
Melnikov 1971). These systems classify LD50's (median lethal dose) into
categories of relative toxicity from "highly toxic" (<50 mg/kg) to
"practically nontoxic" (>5000 mg/kg) with class divisions arranged
geometrically. We developed a similar system for rating subacute data
in which five toxicity classes were recognized. The classes are:
I, <41 ppm; II, 41-200 ppm; III, 201-1000 ppm; IV, 1001-5000 ppm; and
V, >5000 ppm.
Table 2 illustrates differences in general order of toxicity
among chemical classes for Japanese quail, mallards, and rats. The
rat data were derived from previously published acute toxicity tests
(Gaines 1960, 1969; Melnikov 1971).
Japanese quail and mallards showed similar responses, except that
mallard results fell slightly more often into the least toxic class, V.
Both species responded similarly to organochlorine and organophosphorus
compounds. These chemical classes tended to be most toxic to birds, as
they contained the greatest proportion of compounds rated class I and II.
All carboxylates and most "miscellaneous" compounds fell in the least
toxic class. The toxicity ratings for rats were different because a
much higher percentage of compounds were judged to be highly toxic
(classes I and II) .
With the possible exception of carboxylates, it is clear that lethal
hazard cannot be predicted solely on the basis of chemical class. Nor
can it be assumed that relative hazard of chemicals based on acute study
with rats will follow the same order when tested subacutely on birds.
In Table 3 relative toxicities of different classes of chemicals
to birds are given in more detail by subdividing the major classes
according to their structural properties. Here, toxicities of similar
compounds follow more definable patterns.
Organochlorines tested are halogen derivatives of either alicyclic
or aromatic hydrocarbons. Nearly all compounds in class I and II are
alicyclic. In contrast, most aromatic hydrocarbons are in class IV or V.
There are exceptions, however. For example, Starlicide, an aromatic
hydrocarbon bird control agent, is surpassed in subacute toxicity to
Japanese quail only by endrin, an alicyclic hydrocarbon, and azodrin, an
organophosphorus compound (Table 1) .
Tests with rats followed a similar toxicity pattern (Gaines 1960,
1969). Alicyclic hydrocarbons generally produced LD50's of less than
100 mg/kg (class I and II) whereas aromatic hydrocarbons were above 800
mg/kg (class III-V) . Starlicide was generally in line with other aromatics
in rat tests as the reported LD50 is 1170 mg/kg (Christensen 1973) .
Organophosphorus compounds are derivatives of four phosphorus acids:
phosphoric, thiophosphoric, dithiophosphoric, and phosphonic. All organo-
phosphates that fell into class I were derivatives of either phosphoric or
thiophosphoric acids, except Mocap (bobwhite LC50, 33 ppm) , a dithio-
phosphoric acid. Phosphoric and thiophosphoric acids produced few LC50's
above 1000 ppm (class IV and V), although nearly all LC50's of dithio-
phosphoric and phosphonic acids were over 1000 ppm. Among the phosphoric
acids, azodrin (the most toxic compound tested), its close relative
Bidrin, and phosphamidon were consistently among the most toxic of all
compounds tested. Several thiophosphoric acids — Dasanit , famphur,
fenthion, methyl parathion and thionazin — also were highly toxic. In
general, the order of toxicity of these phosphorus acids to birds was:
phosphoric _> thiophosphoric > dithiophosphoric _> phosphonic. Phosphoric
and thiophosphoric acids also were highly toxic to rats (Gaines 1960,
1969). Abate, a thiophosphoric acid, provides an interesting contrast
because it was quite toxic to birds (Table 1 and Hill 1971), but not to
rats (Gaines 1969) .
The metallic compounds tested varied widely in chemical composition
and permitted only general comparisons. Organic forms tended to be more
toxic than inorganic forms. The organomercurials Ceresan M and Morsodren
consistently gave LC50's less than 100 ppm (class II). LC50's for other
metallics usually exceeded 1000 ppm (class IV and V) .
Comparative Toxicity in Relation to Species
Comparisons of the susceptibility of Japanese quail, ring-necked
pheasants and mallards to different chemicals are shown in Table 4. Quail
appeared to be most sensitive to the comparatively toxic alicyclic hydro-
carbons and mallards the least sensitive. Pheasants were most sensitive
to aromatic hydrocarbons and mallards appeared least sensitive (based on
response rating and median LC50) . Species sensitivity to organophosphorus
compounds followed the order: quail > pheasant > mallard. For carbamates,
the order was quail > mallard > pheasant. This indicates a change in the
pheasant-mallard relationship, even though organophosphates and carbamates
both are chlorinesterase inhibitors. Meaningful comparisons were not
possible for other classes of chemicals, but some observations are
pertinent. Only pheasants produced LC50's less than 5000 ppm for
carboxylates. Pheasants appeared most sensitive and mallards least
sensitive to inorganic metallics, but the reverse was true for organic
metallics.
Overall comparisons showed the probable order of sensitivity to be
Japanese quail > ring-necked pheasant > mallard; this relationship
occurred 31°' of the time. The opposite order occurred least frequently
(6%). The second most frequent order of sensitivity (27%) was pheasant >
quail > mallard.
Because all possible variations in order of species sensitivity
occurred in all chemical classes, it is clear that an LC50 for any of
these species probably would not permit prediction of that chemical's
toxicity to either untested species. A similar conclusion was reached
by Tucker and Haegele (1971) from the results of tests of acute
pesticidal toxicity to six species of birds.
Although accurate prediction of the sensitivity of one species to a
given compound from data for a different species appears unlikely, there
are positive correlations between LC50's for different species within a
given chemical class. Table 5 shows correlation coefficients for paired
LC50's for the species we tested within the major chemical classes. All
correlations of LC50's for Japanese quail and mallards were statistically
significant (P<0.05 or P<0.01). Correlations between LC50's for ring-
necked pheasants and either mallards or Japanese quail were significant
in three of four comparisons with each species. Only three of eight
comparisons between bobwhite and other species proved significant.
Because all correlations are positive, 17 of 20 are either significant
or very nearly significant at P=0.05, and one-half are highly
significant (P<0.01), it is clear that the test species responded
similarly, in a relative sense.
CONCLUSIONS
We have measured the dietary susceptibility of two to four species
of birds to 131 toxic compounds. From the data provided in Table 1 the
toxicities of different compounds to the same species can be tested
statistically; thus, toxicity rankings are possible.
Most of the more toxic compounds were halogen derivatives of
alicyclic hydrocarbons, derivatives of phosphoric and thiophosphoric
acids, and organomercurials. Carbamates, often of extreme acute toxicity
to rats, were only moderately toxic when fed to birds. Most carboxylates,
ketones, organonitrogen compounds, organosulfates and ureas were of a
relatively low order of toxicity.
Interspecies comparisons showed the overall order of susceptibility
to be quail > pheasant > mallard which is size related. All combinations
of species order occurred. Although the order of susceptibility of species
varied, a characteristic order usually prevailed within a given class of
chemicals and the LC50's for any two test species were strongly correlated.
This suggests that, regardless of test species, the relative toxicities of
different chemicals in the same class would be similar if test conditions
were constant. However, unpredictable differences in species' suscepti-
bility occur often enough that tests of at least two species are desirable.
ACKNOWLEDGMENTS
We are grateful to the many persons who contributed to this study.
Special recognition is extended to the following: William H. Stickel was
an invaluable source of information relating to our selection of test
compounds. Clyde Vance supervised aviculture and consistently provided
high quality test birds on schedule. Marshall Hynson, David Jaquith,
Norman Kruhm, Claude Mills (deceased), Frank Polak, David Prevar, Richard
Rowlett, and Perry Waters serviced test facilities and assisted with data
gathering. William L. Reichel, T. Earl Kaiser and Thair G. Lamont advised
on various chemical considerations. Helen L. Young conducted the computer
analyses. Lucille F. Stickel and J. Larry Ludke critically reviewed the
manuscript. Special thanks go to the various chemical companies who
provided the compounds for testing.
REFERENCES
Bliss, C. I. 1952. The statistics of bioassay. Pages 445-628 in Vitamin
methods, Vol. II. Academic Press, New York.
Casewell, R. L. , D. E. Johnson, and C. Fleck. 1972. Acceptable common
names and chemical names for the ingredient statement on pesticide labels,
2d ed. U. S. Environ. Prot. Agency, Washington, D.C. 243 pp.
Christensen, H. E. , editor. 1973. The toxic substance list. U. S. Dep.
Health, Educ, Welfare, Rockville, Md. 1001 pp.
Daum, R. J., and W. Killcreas. 1966. Two computer programs for probit
analysis. Bull. Entomol. Soc . Am. 12 (4) :365-369.
Finney, D. J. 1952. Probit analysis, 2d ed. Cambridge Univ. Press.
318 pp.
Finney, D. J. 1964. Statistical method in biological assay, 2d ed.
Hafner Publ. Co., New York. 688 pp.
Gaines, T. B. 1960. The acute toxicity of pesticides to rats. Toxicol.
Appl. Pharmacol. 2(l):88-89.
Gaines, T. B. 1969. Acute toxicity of pesticides. Toxicol. Appl.
Pharmacol. 14 (3) :515-534.
Heath, R. G., J. W. Spann, E. F. Hill, and J. F. Kreitzer. 1972. Compara-
tive dietary toxicities to birds. U. S. Fish Wildl. Serv. , Spec. Sci.
Rep. Wildl. 152. 57 pp.
Hill, E. F. 1971. Toxicity of selected mosquito larvicides to some common
avian species. J. Wildl. Manage. 35 (4) : 757-762.
Hodge, H. C, and J. H. Sterner. 1949. Tabulation of toxicity classes.
Am. Ind. Hyg. Assoc. Quart. 10(4):93-96.
Litchfield, J. T. , Jr., and F. Wilcoxon. 1949. A simplified method of
evaluating dose-effect experiments. J. Pharmacol. Exp. Ther. 96(2):
99-113.
Melnikov, N. N. 1971. Chemistry of pesticides. Residue Rev. 36:1-480.
Radeleff, R. D. 1964. Veterinary toxicology. Lea and Febiger,
Philadelphia. 314 pp.
Snedecor, G. W. , and W. G. Cochran. 1967. Statistical methods, 6th ed.
Iowa State Univ. Press, Ames. 593 pp.
Tucker, R. K. , and M. A. Haegele. 1971. Comparative acute oral toxicity
of pesticides to six species of birds. Toxicol. Appl. Pharmacol. 20
(1): 57-65.
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Table 2. Percentage frequency distribution by toxicity class for pesticidal
compounds tested subacutely against Japanese quail and mallards
as compared to acute rat toxicities .
Chemical class
Species
Toxicity class
II
III
IV
V
Organochlorine
Quail
(n = 18)
Mallard
Rat
Organophosphorus
Quail
(n = 35)
Mallard
Rat
Carbamate
Quail
(n = 12)
Mallard
Rat
Carboxylate
Quail
(n = 7)
Mallard
Rat
Metallic
Quail
(n = 9)
Mallard
Rat
Miscellaneous
Quail
(n = 14)
Mallard
Rat
11
11
6
11
11
33
9
26
6
6
43
31
8
33
25
14
22
22
22
56
21
33
33
11
28
28
28
33
6
17
31
23
11
40
26
23
17
6
3
25
33
42
25
33
33
42
100
100
43
29
14
11
56
11
11
11
56
22
14
7
79
7
21
71
50
29
Only compounds providing complete data for each species are included. Avian data
are from Table 1; rat data are from Gaines (1960, 1969) or Melnikov (1971).
^Bounds of toxicity ratings: I = <41, II = 41-200, III = 201-1000, IV = 1001-
5000, V = >5000. Toxicologic parameters are LC50's as ppm compound in diet of
birds and LD50's as mg compound per kg body weight for rats.
"Miscellaneous includes nitrophenol, ketone, organonitrogen, organosulfer and
urea compounds.
37
Table 3. Frequency distribution by toxicity class for organochlorine,
organophosphorus , carbamate and metallic compounds tested subacutely
against birds .
Species
Toxicity
i c
class
Chemical class
I
II
III
IV
V
ORGANOCHLORINE COMPOUNDS
Derivatives of
alicyclic hydrocarbons
BW
3
1
5
1
JQ
2
2
5
1
1
PH
1
2
5
3
ML
1
2
4
2
2
Derivatives of
aromatic hydrocarbons
BW
5
6
3
IQ
1
1
9
6
PH
4
9
3
ML
10
7
ORGANOPHOSPHORUS COMPOUNDS
Derivatives of
phosphoric acid
BW
1
1
JQ
2
2
2
1
1
PH
1
2
2
1
1
ML
1
1
1
3
1
Derivatives of
thiophosphoric acid
BW
2
6
3
JQ
7
5
1
1
PH
5
7
2
ML
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2
8
1
2
Derivatives of
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BW
1
4
1
JQ
1
4
5
3
PH
1
4
5
3
ML
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5
4
Derivatives of
phosphonic acid
BW
JQ
PH
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2
2
1
2
3
ML
1
2
1
38
Table 3. Frequency distribution by toxicity class for organochlorine,
organophosphorus , carbamate and metallic compounds tested subacutely
against birds — continued
Chemical class
Species
Toxicity class
II
III
IV
CARBAMATE COMPOUNDS
Derivatives of
carbamic acid
BW
JQ
PH
ML
1
3
3
3
2
5
6
6
1
3
3
2
Derivatives of
dithiocarbamic acid
BW
JQ
PH
ML
1
1
2
3
3
3
METALLIC COMPOUNDS
Inorganic
JQ
PH
ML
3
2
1
1
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Organic
BW
JQ
PH
ML
1
2
2
2
1
4
4
1
1
3
Basis for frequency distribution is Table 1.
Species: BW, bobwhite; JQ, Japanese quail; PH, ring- necked pheasant; and,
ML, mallard.
"Bounds of toxicity ratings, I = <41 ppm, II = 41-200 ppm, III = 201-1000 ppm,
IV = 1001-5000 ppm, and V = >5000 ppm.
39
Table 4. Comparative responsiveness among young Japanese quail, ring-
necked pheasants and mallards to pesticidal compounds when
tested subacutelya.
Chemical class
Species
Response rating" Median
Most
Least
LC5Q
(Extremes)
ORGANOCHLORINE
Derivatives of alicyclic hydrocarbons
(10)d
Derivatives of aromatic hydrocarbons
(15/12)
Quail
50%
0
388
( 18->5000)
Pheasant
30%
20%
495
( 14- 1540)
Mallard
20%
80%
669
( 22->5000)
Quail
17%
50%
3165
( 568->5000)
Pheasant
84%
8%
2078
( 311->5000)
Mallard
0
42%
3572
(1651->5000)
Quail
67%
0
286
(
2->5000)
Pheasant
33%
0
246
(
3->5000)
Mallard
0
100%
1991
(
32->5000)
ORGANOPHOSPHOROUS
Derivatives of phosphoric acid
(7/6)
Derivatives of thiophosphoric acid
(13)
Derivatives of dithiophosphoric acid
(12/10)
Derivatives of phos phonic acid
Quail
69%
8%
211
(
47->5000)
Pheasant
15%
23%
240
(
49->5000)
Mallard
15%
69%
640
(
43->5000)
Quail
50%
15%
2067
( 100->5000)
Pheasant
40%
20%
2230
( 118->5000)
Mallard
10%
65%
3300
( 248->5000)
(4)
Quail
25%
0
972
( 295- 1901)
Pheasant
50%
25%
708
( 270- 3401)
Mallard
25%
75%
1430
(~330->5000)
40
Table 4. Comparative responsiveness among young Japanese quail, ring-
necked pheasants and mallards to pesticidal compounds when
tested subacutely3 — continued
Chemical class
Species
Response rating" Median
Most
Least
LC-rt
(Extremes)
CARBAMATE
Derivatives of carbamic acid
(11/9) Quail
Pheasant
Mallard
56%
22%
1427
( 381->5000)
11%
67%
2874
( 573->5000)
33%
11%
2300
( 190->5000)
Comparisons are restricted to compounds providing comparable data among
the species as shown in Table 1. Mallard values used are mainly for
10-day-old birds.
Percentage of times each species produced the lowest (most responsive)
or highest (least responsive) LC50 for compounds within each chemical
class.
c Derivation of median toxicities were restricted to LC50's of compounds
used in construction of this table.
Where two numbers are shown, the first represents total compounds used
for determination of median toxicities and the second is the total
compounds upon which response rating percentages are based.
41
Table 5. Relation between four avian species in subacute responsiveness to pesticidal
compounds3.
Chemical class
Species compared
Statistic
OC
OP
CB
IMfOM
Bobwhite - Japanese quail n
r
Bobwhite - ring-necked pheasant n
r
Bobwhite - mallard n
Japanese quail - ring-necked n
pheasant
r
Japanese quail - mallard n
Ring-necked pheasant - mallard n
5 13 6
0.704 0.900** 0.491
I.D. 11
0.561
6 13
0.759 0.543
12 30
I.D.
6 I.D.
0.675**
6 I.D.
0.958**
11 7
0.679* 0.941** 0.486 0.836*
14
28
11
11
0.918** 0.853** 0.633* 0.774**
12
28
13
0.897** 0.902** 0.902** 0.622
Correlation coefficients are for paired LC50's from standardized data in Table 1.
Standardized data includes only results for tests of 12-18-day-old quail and 8-12-
day-old pheasants and mallards.
n, number of paired LC50's; r, correlation coefficient; I.D., insufficient data.
C0C, organochlorine; OP, organophosphorus ; CB, carbamate; IM, inorganic metallic; OM,
organometallic.
*Correlation coefficient statistically significant (P<0.05).
**Correlation coefficient highly significant (P<0.01).
42
APPENDIX
43
Appendix 1
Toxicity Statistics
The principal statistical reference point is the LC50, as determined
by computerized probit analysis. The LC50, as used under our procedure,
is ppm toxicant (based on active ingredient) in an ad libitum diet produc-
ing 50% mortality in 8 days (5 days of toxic diet followed by 3 days of
untreated diet).
The probit analysis program calculates the following maximum likeli-
hood statistics: LC50 and its 95% confidence limits; slope of the
weighted linear regression of probits on log-concentration and its
standard deviation; and relative toxicity, with 95% confidence limits, of
any two compounds after testing regression lines for parallelism and
heterogeneity. The program permits simultaneous analysis of all compounds
tested in any single experiment.
Comparison of toxicities between compounds is by determination of
their relative toxicity or "toxicity ratio." The toxicity ratio may be
expressed unconditionally as the ratio between LC50's of two compounds
provided the level of tolerance of test populations is the same and
probit regression lines are parallel. The level of tolerance can be
assumed comparable only if both test populations are drawn from the same
population and are tested concurrently in a completely randomized exper-
iment. Because this condition is obviously restrictive, adjustment for
tolerance differences between experiments is possible with the positive
control, according to the procedure presented in Appendix 2. Parallelism
is assumed if slopes of regression lines are not shown to be different at
a specified level of significance.
Lethal concentrations other than the LC50 may be useful. These
values can be estimated from data in Table 1 by the procedure described
in Appendix 2. Estimates of this type should be derived from especially
designed experiments, however, because extrapolation from a standard
probit regression line can be misleading if the true regression equation
has some curvature (Finney 1952) .
44
Appendix 2
Calculation of Some Significant Toxicity Values
Toxicity ratios: The RTD values listed in Table 2 are used to
calculate the toxicity ratio of two compounds for a particular species
as follows:
1. Compute the toxicity ratio of "Compound 1" to "Compound 2" by
dividing the RTD of Compound 2 by the RTD of Compound 1. For
example, if dieldrin is 4 times as toxic as Compound 1 (RTDi =
4) and 6 times as toxic as Compound 2 (RTD2 = 6) , then
Compound 1 is 1.5 times as toxic as Compound 2 (i.e., RTD2/
RTD j = 6/4 =1.5). An algebraic argument for the procedure was
previously presented (Heath et al. 1972). The calculation of
confidence limits for potency ratios require more data than
could reasonably be included in this paper.
2. Test the slopes of the probit regression lines of the two
chemicals for parallelism using a 2-tailed t-test. Let b and
b2 be the estimated slopes and Sj and s2 their standard devia-
tions. (The s values are actually in standard error form.)
Also let n1 and n2 equal the number of concentrations used in
the respective determinations.
Then
t = (bi - b2) / J71
2 + s22
Since s 2 and s 2 have n1 - 2 and n - 2 degrees of freedom,
t is given n-. + n2 - 4 degrees of freedom, provided si2 and
9 9
s„ can be considered estimates of a common a . We expect
homogeniety of variances in most instances; however, procedures
for testing the equality of two variances and the significance
of the difference of two means (i.e., b^ and b2) when variances
are unequal are presented in Snedecor and Cochran (1967) .
LC's for the percentage of response:
Lethal concentrations for percentages of mortality other than the
median can be estimated from the data in Table 2 as follows:
1. Transform the LC50 to its common logarithm and the desired
percentage of mortality to its probit, the probit of 50% being
5. If we let k equal the new percentage of mortality for which
we wish to estimate the lethal dietary concentration (i.e., the
LC ), and b equal the particular slope value from Table 2, then
k
log LC = log LC50 + (probit k -5) /b .
K.
45
The antilog of LCk is the desired estimate. Tables for transforming
percentages to probits can be found in various statistical texts, including
Finney (1952, 1964).
46
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61
As the Nation's principal conservation agency, the Department of the
Interior has responsibility for most of our nationally owned public
lands and natural resources. This includes fostering the wisest use of
our land and water resources, protecting our fish and wildlife, preserv-
ing the environmental and cultural values of our national parks and
historical places, and providing for the enjoyment of life through out-
door recreation. The Department assesses our energy and mineral
resources and works to assure that their development is in the best
interests of all our people. The Department also has a major responsi-
bility for American Indian reservation communities and for people who
live in island territories under U.S. administration.
GPO 832 - 387
UNITED STATES
DEPARTMENT OF THE INTERIOR
FISH AND WILDLIFE SERVICE
WASHINGTON. D. C 20240
POSTAGE AND FEES PAID
U.S. DEPARTMENT OF THE INTERIOR
INT 423
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