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This is to certify that the

thesis entitled

THE EFFECT OF CHRONIC DIETARY ADMINISTRATION
OF FLURIDONE ON SELECTED REPRODUCTIVE
PARAMETERS OF BOBWHITES AND MALLARDS

presented by

CHRISTINE FLAGA

has been accepted towards fulfillment
of the requirements for

M.S. 4egreein ANIMAL SCIENCE

 

Major professor

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OVERDUE FINES:
25¢ per day per Item

RETURNING LIBRARY MATERIALS:

Place In book retum to remove
charge from circulation records

 

ngIjI 03999

 

 

THE EFFECT OF CHRONIC DIETARY ADMINISTRATION
OF FLURIDONE ON SELECTED REPRODUCTIVE

PARAMETERS OF BOBWHITES AND MALLARDS

BY

Christine Flaga

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE

Department of Animal Science

1981

ABSTRACT
THE EFFECT OF CHRONIC DIETARY ADMINISTRATION

OF FLURIDONE ON SELECTED REPRODUCTIVE
PARAMETERS OF BOBWHITES AND MALLARDS

BY

Christine Flaga

Four dietary concentrations of fluridone (l-methyl-B-
phenyl-S-[B-(triflouromethyl-pheny1Jv4(l§)-pyridinone) were
fed to Bobwhites and Mallards to evaluate its effect on food
consumption, body weight gain, and several reproductive
parameters. Fluridone was fed at 0 ppm (control), 100 ppm
(0.01%), 300 ppm (0.03%), and 1,000 ppm (0.10%) for approxiv

mately six months.

Chronic dietary administration of fluridone to Bobwhites
and Mallards did not significantly affect mortality, food
consumption, body weight gain, egg production, fertility,
embryo survival, hatchability, offspring survivability, or

eggshell thickness.

ACKNOWLEDGMENTS

I would like to express my sincere appreciation to all
my committee members, Dr. Robert K. Ringer, Dr. Steven J.
Bursian, Dr. Richard J. Aulerich, and Dr. Lee R. Shull, for
their unique contributions toward the completion of this
thesis.

I would also like to thank all those who contributed
their technical assistance during the actual course of the
study, especially Mr. William J. Breslin and Dr. Steven J.
Bursian.

Special thanks go to Mr. Terrance Kavanagh for his
invaluable support and friendship and to my father, Mr.

Edward F. Flaga, for his love and encouragement.

TABLE

LIST OF TABLES. . . . .
LIST OF FIGURES . . . .

LIST OF APPENDICES. . .

INTRODUCTION. . . . . .
REVIEW OF LITERATURE. .
OBJECTIVES. . . . . . .
MATERIALS AND METHODS .
RESULTS . . . . . . . .
DISCUSSION. . . . . . .

CONCLUSIONS . . . . . .

APPENDICES. . . . . . .

LITERATURE CITED. . . .

OF CONTENTS

iii

Page
iv
vi

vii

13
‘14
23
64
78

79

90

Table

1.

10.

ll.

12.

13.

14.

LIST OF TABLES

General toxicity of fluridone to birds, fish,
and an invertebrate. . . . . . . . . . . .

Acute mammalian toxicity data for fluridone. .
Interim deaths for Mallards fed fluridone. . .

The effect of chronic dietary administration
of fluridone to Mallards upon feed consump-
tion (grams/bird/day : S.D.) . . . . . . . . .

The effect of chronic dietary administration
of fluridone to male Mallards upon mean per-
cent change of initial body weight (: S.D.). .

The effect of chronic dietary administration
of fluridone to female Mallards upon mean per-
cent change of initial body weight (: S.D.). .

The effect of chronic dietary administration
of fluridone to Mallards upon egg production
(eggs/hen/day : S.D.). . . . . . . . . . . . .

The effect of chronic dietary administration
of fluridone to Mallards on mean reproductive
parameters (: S.D.) for all sets combined. . .

Mallard reproductive parameters expressed as a
percentage of eggs laid. . . . . . . .

Pathology report for the Mallards. . . . . . .
Interim deaths for Bobwhites fed fluridone . .

The effect of chronic dietary administration
of fluridone to Bobwhites upon feed consump-

tion (grams/bird/day i S.D.) . - - . - - - - -

The effect of chronic dietary administration
of fluridone to male Bobwhites upon mean per-
cent change of initial body weight (i S.D.). .

The effect of chronic dietary administration

of fluridone to female Bobwhites upon mean per-
cent change of initial body weight (: S.D.). .

iv

Page

9

10

24

25

29

32

36

41

42

44

51

Table Page

15.. The effect of chronic dietary administration
of fluridone to Bobwhites upon egg production
(eggs/female/day : S.D.). . . . . . . . . . . . 54

16. The effect of chronic dietary administration
of fluridone to Bobwhites on mean reproductive
parameters for all sets combined. . . . . . . . 57

17. Bobwhite reproductive parameters expressed as
a percentage of eggs laid . . . . . . . . . . . 60

18. Pathology report for the Bobwhites. . . . . . . 61

19. Classification systems used to rate the
toxicity of chemicals . . . . . . . . . . . . . 68

20. Classification system used to rate or compare
the subacute dietary LCSOs of chemicals . . . . 69

21. Subacute dietary LCSOs of certain insecticides
and herbicides administered to Bobwhites. . . . 71

22. Acute LDSOs (mg/kg) of certain herbicides and
insecticides administered orally to Mallards. . 72

23. Amount of fluridone (mg/kg/day) ingested by
Mallards. C O O O O O O O O O O O O O O O O O 73

24. Amount of fluridone (mg/kg/day) ingested by
Bobwhites . . . . . . . . . . . . 73

Figure
l. Chronology of study for Bobwhites fed fluridone
2. Chronology of study for Mallards fed fluridone
3. Example of reproductive parameters displayed in
a continuum. . . . . . . . . . . . . . . . . .
4. The effect of chronic dietary administration of
fluridone to Mallards upon mean food consumption
5. The effect of chronic dietary administration of
fluridone to male Mallards upon mean percent
change of initial body weight. . . . . . . . .
6. The effect of chronic dietary administration of
fluridone to female Mallards upon mean percent
change of initial body weight. . . . . . . . .
7. The effect of chronic dietary administration of
fluridone to Mallards upon mean egg production
8. Mallard reproduction parameters expressed in a

10.

11.

12.

13.

LIST OF FIGURES

continuum as a percent of the number of eggs
laid I O O O O O O O O O O O O O O O O O I O O

The effect of chronic dietary administration of
fluridone to Bobwhites upon mean food consump-
tion 0 O O O O O C O O O O O O O O O O O O O O

The effect of chronic dietary administration of
fluridone to Bobwhite males upon mean percent
change of initial body weight. . . . . . . . .

The effect of chronic dietary administration of
fluridone to Bobwhite females upon mean percent
change of initial body weight. . . . . . . . .

The effect of chronic dietary administration of

fluridone to Bobwhites upon mean egg production.

Bobwhite reproductive parameters expressed in a
continuum as a percent of the number of eggs
laid I 0 O O O O O O o o O O I O O I O O O 0 0

vi

Page
16
17

21

28

31

34

38

4O

47

50

53

63

LIST OF APPENDICES

Appendix
A. Structure and solubilities of fluridone. . .
B. Layout of Bobwhite testing room. . . . . . .
C. Layout of Mallard testing room . . . . . . .
D. Composition of adult Mallard mash. . . . . .
E. Calculated analysis of adult Mallard mash. .
F. Composition of Mallard duckling mash . . . .
G. Calculated analysis of Mallard duckling mash
H. Composition of adult Bobwhite mash . . . . .
I. Calculated analysis of adult Bobwhite mash .
J. Composition of Bobwhite chick mash . . . . .
K. Calculated analysis of Bobwhite chick mash .

vii

79
80
81
82
83
84
85
86
87
88
89

INTRODUCT I ON

The U.S. Environmental Protection Agency (EPA) has
proposed, under the Federal Insecticide, Fungicide, and
Rodenticide Act (FIFRA), that avian reproduction studies be
required to support the registration of a formulated pesti-
cide if there exists any suggestion of bioaccumulation or
chronic exposure to the avian species, especially during
the breeding season (Federal Register, 1978). Mallards
and Bobwhites, which are examples of a waterfowl and an
upland game avian species, respectively, were indicated
as test animals of choice.

This particular study was performed to determine the
potential reproductive toxicity of chronic dietary exposure
of fluridone to Bobwhites and Mallards. No such data had been
available previously. Fluridone is an experimental herbi-

cide produced by Eli Lilly and Company (Elanco).

LITERATURE REVIEW

l-methyl-B-phenyl-S-[3-(trifluoromethyl)phenyl]—4(1g)-
pyridinone (fluridone) is an experimental herbicide developed
by Eli Lilly and Company, Indianapolis, IN for use in aquatic
plant management and in cotton fields. It is a white, odor-
less, crystalline solid with a molecular weight of 329.3.
Fluridone melts at 151-1540C and is soluble in water at
12 ppm. See Appendix A for its chemical structure and a
list of its solubilities in organic solvents. Fluridone
has an n-octanol/water partition coefficient of 57.5 (log

7

K = 1.76) and a vapor pressure of l x 10- mm Hg at 25°C

OW
(Lilly, 1978).

Analytical Procedures

 

Methods for residue analysis from soil and plant tis-
sue are discussed by West (1978).

West and Burger (1980) have described a gas liquid
chromatographic method for determining the residues of
fluridone and its major metabolite (l-methy1-3-(4-hydroxy-
pheny1)-5-[3-(trifluoromethyl)pheny1]-4-(l§)-pyridinone) in
fish. Bioassay and gas chromatographic procedures were
evaluated for their effectiveness in detecting fluridone
from soil (Banks et a1., 1979).

Herbicidal Properties

 

Waldrep and Taylor (1976) initially evaluated fluridone
for its pre- and postemergence herbicidal efficacy on several
weed species in cotton. They found it to be herbicidally

active at low dosages for weeds, but cotton was relatively

resistant to its herbicidal action. Its herbicidal activity
was greater when applied preemergence, controlling a wide
variety of annual grasses and broadleaf weeds. Susceptible
plants treated preemergence with fluridone emerged with
chlorotic leaves which became necrotic, subsequently leading
to plant death. The authors characterized fluridone as a
slow-acting, translocated herbicide which appeared to inhibit
chlorophyll synthesis. Specifically, fluridone inhibits
carotenoid synthesis (Bartels and Watson, 1978) leading

to chloroplast photodestruction and the observed loss of
chlorophyll.

Plants susceptible to fluridone translocate the herbi-
cide readily into the shoots (Berard et al., 1978). Cotton
tolerates the herbicide primarily because of fluridone's
limited translocation in this plant species. Any absorbed
fluridone is retained in the roots and basal region of the
stem. Root and shoot tissues from sago and Richardson
pondweed concentrated fluridone with limited root to shoot
translocation but negligible shoot to root transport (Marquis
et al., 1981).

Sanders et al. (1981) reported a reduction in hydrilla
biomass 84 days after treatment with 2 1.7 kg of fluridone/
ha, while insufficient control occurred in plots treated at
0.84 kg/ha.

Persistence in soils.

 

Fluridone strongly adsorbs to organic matter in soils.

Soil microorganisms do not appear to be a major factor in its

dissipation. In cotton-producing areas, residues of fluridone
may carry over to the next planting season causing slight
injury to sorghum, soybeans, sugar beets, and tomatoes that
follow in rotation (Weed Science Society of America, 1979).

Banks and Merkle (1979) reported that fluridone did not
leach more than 1 em in clay or sandy loam soils when up to
10 cm of water was paSsed through a soil column containing
fluridone. Greater downward mobility (12 to 17 cm) occurred
in a coarse sand when 5 or 10 cm of water was passed through
a soil column.

Banks et a1. (1979) determined the residue of fluridone
in Miller clay and Lufkin fine sandy loam at various inter-
vals after incorporated and nonincorporated field application.
Incorporation increased fluridone‘s persistence in Lufkin
fine sandy loam but not in Miller clay. Miller clay retained
a low level of fluridone after 250 days, while up to 25% re-

mained in Lufkin fine sandy loam after 285 days.

Persistence in Pond Water.

 

In 1979, fluridone‘s use as an aquatic herbicide was
researched by W. R. Arnold of the Lilly Research Laboratories
(Arnold, 1979). Fluridone was applied at various rates as a
surface or bottom treatment to ponds. Little weed control
was noticed until two to four weeks after treatment. Mature
vegetation slowly decomposed and sank to the bottom gradually
increasing the percentage of open water. Applied at 1.68
kg/ha, fluridone provided excellent control of hydrilla,

common elodea, southern naiad, cattail, para grass, and

D.

in

 

nu.

.wi

bu

several other species without adversely affecting phyto-
plankton, benthic organisms, or fish. Bluegills taken from
the treated ponds were analyzed for fluridone residues which
were detectable until 29 days after treatment (0.031 ppm with
application of 1.68 kg/ha). At no time did the residue level
in the fish exceed the concentration in the water. Observa-
tions, before and after treatment, were made of other aquatic
organisms including crayfish, bass, catfish, turtles, frogs,
watersnakes, and waterfowl. No adverse effects were observed
in these species. It was also noted that a variety of water-
fowl and shore birds continued to feed at the treated ponds.
Determination of the fluridone content of water and hydrosoil
indicated that most of the fluridone dissipated from the water
into the hydrosoil. Minor additional dissipation was attri-
buted to uptake by aquatic organisms and photodegradation.
McCowen et a1. (1979) reported similar results when
fluridone was applied to ponds by various methods at rates
reSulting in concentrations of 0.1 to 10 ppm. Fluridone
provided control of many submersed and emersed aquatic
plants. Residue levels were determined in water, hydrosoil,
and fish. At an application rate resulting in a concentration
of 0.1 ppm, fluridone residue levels were determined 54 days
after application at < 0.0005 ppm and at 0.035 ppm in water
and hydrosoil, respectively. One week after treatment,
fluridone levels in water and hydrosoil were 0.021 and 0.220
ppm respectively. The half-life (t 1/2) of fluridone in ponds

was determined to be 14 days or less. Fluridone was not

6

found to accumulate in fish. Mosquito fish (g. affinis) sur-
vived and reproduced at all rates of fluridone application.
No adverse effects upon other aquatic life were observed.

Both of the above studies indicated that fluridone did
not affect water quality parameters such as pH, BOD, color,
dissolved solids, hardness, nitrate nitrogen, total phos-
phates, or turbidity.. Arnold (1979) did report an increase
in the dissolved oxygen concentration but McCowen et a1. (1979)
reported no significant change.

West et a1. (1979) applied fluridone to small ponds in
the U.S. and Gatun Lake in the Panama Canal Zone. Fluridone
dissipated rapidly from the water and the authors credited
it to deposition in hydrosoil and uptake by aquatic plants,
with photolysis as an additional contributing factor. The
half-life of fluridone in pond water averaged 5 days. In
fish, a maximum fluridone residue of 0.054 ppm was determined
one day after treatment (DAT), decreasing steadily to a non-
detectable level on 14 DAT. This data was determined from
subsurface application of fluridone (formulated as a 4 lb./
gallon aqueous suspension) at a rate of 0.1 ppm relative to
the total water column (equivalent to 1.12 kg/ha). The bio—
concentration factor for fish ranged from 0 to 1.7 (determined
as the concentration in fish divided by the concentration in
the water). Bioconcentration was also very low in zooplankton
and aquatic plants.

In another study (Muir et al., 1980), fluridone was

applied to three small ponds at 70, 700, and 5,000 ug/l.

Re.

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Residue analysis was conducted on water, hydrosoil, duck-
weed, and minnows for 70 weeks after application. The half-
life of fluridone in the water column (0.5 m depth) ranged
from 4 (at 700 uq/l) to 7 days (70 ug/l). Results

indicated that fluridone has a half-life of one year or more
in the hydrosoil (at all treatment levels). Fluridone resi-
due levels in minnow tissue ranged from < 0.02 to 0.14 ug/g
(wet weight) throughout the sampling period, with bioconcen-
tration factors ranging from 0 to 64. Fluridone levels in
duckweed were proportional to the herbicide concentrations
in the pond water with bioconcentration factors ranging from
19 to 85.

Other dissipation information was reported by West and
Parka (1981). Fluridone was applied at 0.84 kg/ha of surface
water to the surface and bottom of ponds. Half-lives in the
water column were determined to be 21 and 26 days (surface
and bottom application, respectively). No detectable resi-
due remained in the hydrosoil 56 days after treatment by
either method of application. Similar water and hydrosoil
dissipation results were observed by Sanders et a1. (1981).
They stated that less than 15% of the applied compound
remained after 56 days. Fluridone had been applied at levels
of 0.84, 1.00, 1.70, 3.36, and 6.70 kg/ha to hydrilla test
plots. No adverse effects were observed on dissolved oxygen
or other water quality parameters nor were there any notice-

able disturbances to the plankton and benthic communities.

 

C1

We

Toxicity of Fluridone.

 

Toxicity_to wildlife and fish: Fluridone-contaminated
diets were administered to lG-day-old Mallard ducklings and
lO-day-old Bobwhite chicks for a period of five days. LCO
(maximum nonlethal conc.) values were calculated from the data
produced during these studies. Acute LDOs for Bobwhites and
Mallards were calculated by administering a range of single
doses of fluridone. 96-hour static toxicity tests were con-
ducted on bluegills and rainbow trout. Similar tests were

conducted with Daphnia magna for 48 hours. Fathead minnows

 

were used to determine a maximum acceptable toxicant concen-
tration. Data from these studies are reported in Table 1.

Acute toxicity: Fluridone was administered to mice

 

and rats as an oral or subcutaneous single dose. An aqueous
suspension formulation containing 45% fluridone was also
administered as a single oral dose to rats. Ocular and dermal
toxicity tests were conducted with rabbits. The rat was
utilized as the test animal in studies conducted to determine
the toxicity of fluridone when inhaled for one hour. Results

of these acute studies are listed in Table 2.

Subchronic toxicity; Fluridone toxicity was evaluated

 

in rats, mice, and dogs for periods of three months. There
were no treatment-related effects when fluridone was admin-
istered to mice at dietary doses of 330 ppm or to rats at
dietary doses of 62 ppm (Lilly, 1981). However, rats and
mice fed 2,000 ppm of fluridone showed slight histological

changes in the liver and kidney (Weed Science Society of

I29”

Table 1. General toxicity of fluridone to birds, fish and
an invertebratel.

 

 

 

 

 

 

 

 

 

 

 

Species Route Toxicity
Mallard Diet (5 days) LC0 > 5,000 ppm
(Anas platyrhynchos)
Bobwhite Diet (5 days) LC0 > 5,000 ppm
(Colinus virginianus)
Bobwhite Oral (acute) LD > 2,000 mg/kg
(Colinus virginianus) 50
Bluegill Water (static) LC50 >9<12.5 ppm
(Lepomis macrochirus)
Rainbow trout Water (static) LC50 11.7 ppm
(Salmo gairdneri)
Daphnia Water (static) ECSO 6.3 ppm
(Daphnia magna)

. ‘)
Fathead minnow A Water MATc“ > 0.48 ppm
(Pimephales promelas) (flow through) < 0.96 ppm

 

(Lilly, 1981)
Maximum Acceptable Toxicant Concentration

10

 

 

 

 

 

 

 

 

 

Table 2. Acute mammalian toxicity data for fluridonel.
Species Material Route Toxicity
Rat Technicalz Oral LD50 >1o,ooo mg/kg
(Rattus
norvegicus)
Technical Subcutaneous LD0 >2,000 mg/k
Technical Inhalation LCO >2,l30 mg/M of air
4 AS Oral LDO >0.5 ml/kg
4 AS Inhalation LC0 >9.6 ml/M of air
Mouse Technical Oral LDSO >10,000 mg/kg
(Mus
musculus) Technical Subcutaneous LD50 >2,000 mg/kg
Cat Technical Oral LD0 >250 mg/kg
(Felis
domesticus)
Dog Technical Oral LDo >500 mg/kg
(Canis
familiaris)
Rabbit Technical Dermal LD >500 mg/kg
(Oryctolagus (n8 irritation)
cuniculus) Technical Ocular Moderate irritant
(44 mg/eye)
4 AS Dermal LD >2m1/kg
(s ight irritation)
4 AS Ocular Very slight irritant

(0.1 ml/eye)

 

1 Lilly, 1981.

Technical grade fluridone.

4 pound per gallon aqueous suspension (AS).

nc
we
dc
ad
cc

.1 a D.

11

America.,l979). Further data relating to these changes are
not currently available (Cochrane, 1981). No toxic effects
were observed when fluridone was administered to dogs at
doses of up to 200 mg/kg/day (Lilly, 1981).

Chronic toxicity: Fluridone-contaminated diets were

 

administered to rats for either one or two years. No toxi-
cological effects were observed at a dietary level of 200 ppm.
There was also no evidence of a carcinogenic effect at this
level. Similar long-term feeding studies were conducted in
mice. No gross toxic effects were observed, although these
studies have not been fully evaluated (Lilly, 1981).

Reproductive toxicity: Fluridone was administered to

 

pregnant rats and rabbits during the organogenesis phase of
gestation. At 200 mg/kg/day (rats) and 750 mg/kg/day (rabbits).
no teratogenic effects were noted. The reproductive effects

of fluridone were evaluated by maintaining three successive
generations of rats on diets containing 2,000 ppm of fluridone.
Fluridone did not produce impairment of fertility, live-born
litter size, gestation length, progeny survival,

or sex distribution. In addition, there was no indication of
teratogenicity (Lilly, 1981).

Mutagenesis: In a modified Ames test, 1,000 ug/ml of

 

fluridone produced no evidence of bacterial mutagenesis. Con-
centrations of up to 1,000 nanomoles of fluridone/ml did not

induce cultured hepatocyte DNA repair synthesis. Administered
as an oral dose of 2,000 mg/kg to male rats, fluridone did not

produce dominant lethal mutations (Lilly, 1981).

12

Metabolism.

A detailed investigation of fluridone metabolism in the
rat is currently in progress. Preliminary studies indicate
that a single, oral dose is rapidly absorbed and extensively
metabolized in the rat. The primary route of excretion is
the feces.

The major biotransformation product of fluridone in
fish is l-methyl-3-(4-hydroxyphenyl)-5-[3-trifluroromethyl)
phenyl]-4(1H)-pyridinone. This information was determined
during carbon-l4 fluridone metabolism studies utilyzing fat-

head minnows and bluegills (Lilly, 1981).

Current Status.

 

As of this date, fluridone is not fully registered by
EPA as an aquatic herbicide or for use in cotton, and the
additional existing toxicity and metabolism data are not
available to the general public (Cochrane, 1981).

A temporary tolerance of 0.01 ppm was established for
residues of fluridone in potable water (Federal Register,

1981) and in fish (Federal Register, 1980).

 

To
tic
gaj
To
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tic
sur
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13

OBJECTIVES

To determine the effect of chronic dietary administra-
tion of fluridone on food consumption and body weight
gain of Bobwhites and Mallards.

To determine the effect of the chronic dietary adminis-
tration of fluridone on egg production and fertility of
Bobwhites and Mallards.

To determine the effect of chronic dietary administra-
tion of fluridone on embryo survival, hatchability and
survivability (2 weeks) of the offspring.

To determine the effect of chronic dietary administra-

tion of fluridone on eggshell thickness.

14

MATERIALS AND METHODS

Mallards (Anas platyrhynchos) and Bobwhites (Colinus

 

virginianus) were the two species used as test animals in

 

this avian reproduction study. The Mallards were obtained

from the Max McGraw Foundation, Elgin, IL while the Bobwhites
were supplied by Barrett's Quail Farm of Houston, TX. When
first received, the birds were placed in quarantine for one
week. All birds were then individually banded, placed ran-
domly into their respective testing cages and allowed to ac-
climate for two weeks. In addition, Mallards were wing-clipped
to prevent their escape from the open-topped pens. The Mallards
were lB-weeks-old and the Bobwhites were 16.5-weeks-old when
testing began.

The Mallards and Bobwhites were housed in separate testing
rooms at the Michigan State University (MSU) Poultry Research
and Teaching Facility in East Lansing, MI. In the Mallard
room, two males and five females were randomly assigned to
each testing pen measuring 1.5 m x 1.6 m x 0.7 m (w x l x h).
In the Bobwhite room, one male and one female were randomly
assigned to each testing cage measuring 40 cm x 43 cm x 44.5 cm
(w x l x h). Four pens of Mallards and fifteen cages of
Bobwhites were utilized for each treatment level. Dietary
concentrations of treated feed were randomly assigned to each
pen or cage. Appendices B and C illustrate the testing room
layouts.

Control and treated diets for the adult Mallards, adult

15

Bobwhites, Mallard ducklings, and Bobwhite chicks were pre—
pared by Eli Lilly and Co. The mash-type feed and water were
provided ad libitum to all birds. Diet compositions and
analyses are listed in Appendices D through K. Each feed
shipment, received every six weeks, was sampled and frozen
for future fluridone analysis.

The dietary treatment levels used in this study were
determined from a palatability test conducted at MSU during
the summer of 1979. Some food refusal occurred at a level
of 1.0% (10,000 ppm) fluridone for both Bobwhites (P < 0.05)
and Mallards (P < 0.01). The highest concentration chosen
for this reproduction study (1,000 ppm fluridone) was the
highest level causing no change in food consumption during
the palatability test. The four dietary concentrations used
in this study were: 0 ppm (control), 100 ppm (0.01%), 300
ppm (0.03%), and 1,000 ppm (0.10%) fluridone.

Testing. Treated feed was administered to the birds on
day one of the study's preproduction phase - September 18,
1979 for Mallards and September 19, 1979 for Bobwhites. This
phase ended two months later when the photOperiod was in-
creased from 7 hours 1ight:l7 hours dark to 16 hours light:
8 hours dark. The increased photoperiod stimulated egg
laying thus beginning the study's production phase. Flow
charts of the studies are given in Figures 1 and 2.

Eggs were collected daily when egg production reached
fifty percent for all females within a testing room. Each

Egg collected was marked in pencil with the corresponding

16

Quail received
August 29, 1979

Quarantine
August 29 - Sept. 4, 1979

Acclimation
Sept. 5 - Sept. 18, 1979

Pre-production
Sept. 19 - Nov. 13, 1979

Production
Nov. 14, 1979 - March 2, 1980

Adult terminal kill Egg collection
March 5, 1930 Dec. 17, 1979 - March 2, 1980

Final kill of Zéweek-old
chicks from set #11
May 1, 1980

Figure 1. Chronology of study for Bobwhites fed fluridone.

17

Ducks received
August 23, 1979

 

W
Quarantine
August 23, - Sept. 3, 1979

 

W

Acclimation
Sept. 4 - Sept. 17, 1979

Pre-production
Sept. 18 9 Nov. 12, 1979

 

)

Production
Nov. 13, 1979 - March 13, 1980

Adult terminal kill Egg collection
March 26, 1980 Dec. 17, 1979 - March 13, 1980

1

Final kill of 2-week-old
ducklings (set #10)
April 24, 1980

Figure 2. Chronology of study for Mallards fed fluridone.

18

date and cage number. Prior to this time, any eggs laid
were discarded. Eggs were stored at 15.60C (14.4-16.7OC)
until one week's collection had accumulated. The Mallard
eggs were collected for ten weeks. The Bobwhite collection
period was extended an additional week because a set of eggs
was not transferred from incubator to hatcher.

Eggs (one week's collection) were incubated weekly in a
Jamesway, single stage, 252 incubatorl maintained at dry and
wet bulb temperatures of 37.5°C (37.2-37.80C) and 30°C (29.4-
30.6OC), respectively. Eggs were candled on day 11 for
Bobwhites and on day 14 for Mallards to determine the number
of fertile eggs, infertile eggs, and early dead embryos.
Eggs were candled again on day 18 (Bobwhites) and day 21
(Mallards) of incubation for an interim death determination.
On day 21 (Bobwhites) or day 23 (Mallards) of incubation,
the eggs were transferred to a Jamesway incubator1 with dry
and wet bulb temperatures of 37.20C (34.9-35.50C) and 31.7°C
(31.1-32.20C), respectively. On day 24 for Bobwhites and
day 27 for Mallards, all hatched, pipped, and unpipped off-
spring were removed from the hatcher and their numbers were
recorded. Each hatched bird was individually wing-banded
and recorded according to the parents' dietary fluridone con-

centration.

 

1James Manufacturing Company, Inc. (a subsidiary of Butler

Manufacturing Co.), Fort Atkinson, WI 53538.

19

Hatchlings were transferred from Anthony Hall by auto-
mobile to Petersime 250-24 Battery brooders1 located at the
MSU Poultry Research and Teaching Facility. Temperatures
in the brooders were maintained at 35°C. Starter mash and
water were provided ad libitum. All morbidity and mortality
was recorded daily. At the end of the two-week-period,
surviving offspring were killed by exposure to chloroform.

Feed consumption of the adult birds was measured bi-
weekly throughout the study. Separate feed containers for
each pen or cage facilitated feed weighing. Mallard feed
was weighed to the nearest 10 grams. Bobwhite feed was
weighed to the nearest gram. Body weights were measured
to the nearest gram at 0, 2, 4, 6, and 8 weeks and at ter-
mination. Body weights were not measured during the egg
collection period to avoid any adverse effects caused by
handling.

The adult birds were killed by C02 asphyxiation when
the egg collection period was terminated. At this time, all
birds were necropsied and tissues for histopathology were
taken from at least five animals of each sex per treatment
level. The following tissues were taken: kidney, liver,
heart, lung, spleen, pancreas, proventriculus,gizzard,
duodenum, jejunum, ileum, cecum, colon, testes, ovary,
magnum, shell gland, pectoral muscle, leg muscle, thymus,
sciatic nerve, and skin. Samples collected were placed

in buffered formalin and transported to the Lilly Research

 

1Petersime Incubator Company, Gettysburg, OH 45328.

WE

pC

la

the

WdE

dVE

197E

as;

numb

20

Laboratories for histological processing and examination

by Lilly personnel. Birds that died on test were necropsied
and samples for histopathology were taken and preserved if
the birds displayed any gross pathological abnormalities.

Eggs were collected once every other week for eggshell
thickness measurements. The eggs were cracked in half, the
contents were removed, and the shells were rinsed with tap
water to remove the albumen. The shells were then left to
air dry at room temperature for at least 48 hours. Eggshell
(plus membranes) thickness was measured using an Ames Pocket
Thickness Measurel. Four equidistant points around the
shell circumference were approximated, and the thickness
was measured to the nearest 0.01 mm. From these four data
points, an average for that eggshell was calculated.

The amount of fluridone ingested (mg/kg/day) was calcu-
lated using food consumption and body weight data along with
the dietary concentration. Since food consumption by sex
was not known, body weights were calculated as a total

average with no consideration given to sex.

Statistics.

 

Mallards: Data (except on test mortality) were analyzed
by one-way analysis of variance and a Dunnet's t-test (Gill,
1978a and 1978b). Reproductive parameters were calculated
as percentages of the previous parameter beginning with the

number of eggs laid and continuing through the number of

 

1B. C. Ames Company, 111 Lexington St., Waltham, MA 02154.

('1)

t1

21

the number of offspring surviving 14 days. This method
accounts for all eggs laid by sequentially comparing repro-
ductive parameters. These data, converted to a percentage
of the number of eggs laid, can then be illustrated as a
decreasing continuum. Figure 3 illustrates a hypothetical
example of this continuum and the relationships and defini-
tions of the reproductive parameters.

The on-test mortality data were analyzed by application
of a Bonferroni Chi-Square test to a 2 x 2 contingency table
comparing each treatment level's mortality to the control
(Gill, 1978a and 1978b).

Adjustments were made in duckling survivability when-
ever dead offspring were found without wing bands. In such
cases, the numbers of bandless, dead ducklings were appor-
tioned among the stock cage numbers within their treatment
level for that particular set number.

Bobwhites: The food consumption, body weight, and egg

 

production data were analyzed by one-way analysis of vari-
ance (Gill, 1978a and 1978b). The remaining data were
analyzed by application of a Bonferroni Chi-Square test to
a 2 x 2 contingency table comparing each treatment group

to the control group (Gill, 1978a and 1978b). Information
pertaining to the reproductive parameters and offspring sur-
vivability as stated above under Mallards also applies

to the Bobwhites.

inn

22

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2 3
RESULTS

Mallards. Mallard deaths occurring on test are listed
chronologically in Table 3. Although no significant dif-
ferences were calculated for either males or females, note
that the first six dead birds are all males while the fol-
lowing ten birds are all female. Most interim deaths were
caused by excessive aggressive trauma. The victims were
treaded, cannibalized, and not allowed to feed. Prior to
and at death, birds were devoid of many or most feathers
with hematomas and abrasions located in the skin. Although
body weights of the dead birds were not taken, they all
appeared to have experienced a severe body weight loss.

Most were noticeably moribund prior to death.

Feed consumption of ducks on the fluridone-treated
diets was not significantly different from controls for most
measurement periods (Table 4). A significant difference
(P < 0.05) from the control value was calculated for the
100 ppm treatment group after 18 weeks on treated feed.
However, the control value was lower than the treated value.
A graphic representation of feed consumption over time is
presented in Figure 4.

Analysis of variance revealed no significant differences
for male or female body weight data (Tables 5 and 6, respec-
tively) expressed as mean percent change of initial body
weight. Graphs of the body weight data for male and females

are illustrated in Figures 5 and 6, respectively.

Ta}

di«

12

15.
15

24

 

 

 

Table 3. Interim deaths for.Mallards fed fluridone.

Pen #/ Bird # Date of Cause of
dietary level (sex). death death
12/0.01% 5014 (M) November 16 Excessive aggressive

trauma
5/0-00% 5023 (M) November 18 " " "
2/0-10% 5015 (M) November 19 " " "
3/0-10% 5067 (M) November 29 " " "
5/0-03% 5036 (M) November 30 " " "
10/0.00% 5179 (F) January 14 u n "
5/0-00§ 5259 (F) January 16 " " "
5/0.03% 5203 (F) January 23 n n "
7/0.03% 5122 (F) January 24 " " "
6/0.00% 5304 (F) January 26 " " "
15/0.01% 5275 (F) January 28 " " "
16/0.00% 5273 (F) January 31 Systemic aspergillosis
and excessive aggres-
sive trauma.
9/0.03% 5155 (F) February 3 Excessive aggressive
trauma
15/0.01% 5193 (F) February 18 Bacterial septicemia
and excessive aggres-
sive trauma.
8/0.10% 5408 (F) February 25 Bacterial septicemia

 

*

Cause of death diagnosed by a veterinarian from Eli Lilly

and Co.

Tr.

14

16

18

25

 

 

 

 

Table 4:. The effect of chronic dietary administration of
fluridone to Mallards upon feed consumption
(grams/bird/day : S.D.).

Weeks Dietary treatment (PPm)
on
Treatment 0 100 300 1000
2 115.027 125.0 3 117.0 124.8
(:1; 24.39) (1 30.10% (: 12.19)a (j; 13.99)a
4 128.0 137.0 117.5 133.3
(: 25.81) (: 28.18)a (: 5.07)a (: 18.23)a
6 127.0 143.5 163.5 136.5
(: 15.85) (: 30.36)a (i 51.49)a (: 20.57)a
8 123.3 140.5 123.8 137.3
(1 21.96) (: 27.87)a (: 16.17)a (: 25.67)a
10 79.5 91.5 89.5 86.3
(1 29.72) (: 8.23)a (: 7.14)a (: 16.48)a
12 122.3 133.3 118.0 122.0
(1 17.71) (: 21.70)a (: 16.15)a (: 23.85)a
14 140.0 157.8 141.0 138.0
(: 26.52) (i 21.82)a (i 17.81)a (: 19.87)a
16 164.0 194.3 179.3 169.3
(: 26.88) (: 13.96)a (: 14.89)a (: 19.69)a
18 187.8 233.5 207.0 186.0
(1 28.71) (: 11.24)b (: 22.01)a (: 7.44)a

 

cont'd

male 4

fi
*

Weeks
on
Treatme

 

20

22

24

26

28

26

Table 4 cont' d.

 

 

 

 

Weeks - Dietary treatment (PPm)
on '
Treatment 0 100 300 1000
20 201.5 231.3 202.8 199.5

(1 40.25) (i 11.76)a (i 17.23)a (: 18.36)a

22 198.3 211.3 196.8 192.0
(i 35.52) (: 18.41)a (: 36.51)a (: 23.45)a

24 187.8 200.0 197.3 191.3
(1,25°43) (i 19.10)a (i18.63)a (i 19.33)a

26 163.3 173.8 164.5 163.3
(1 32.77) (: 28.11)a (: 31.94)a (: 22.34)a

28 178.3 188.0 176.0 169.8
(: 26.17) (: 27.39)a (134.86)a (: 25.64)a

 

1Food consumption values represent a biweekly average.

2n = 4 for all means.

3Means with subscript "a" are not significantly different
from control values (P > 0.05); means with subscript "b"
are significantly different from control values (P < 0.05).

27

Figure 4. The effect of chronic dietary administration of
"fluridone to Mallards upon mean food consumption.

28

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mpumaaaz mama ou mcowwuoau mo sowumuumwcweom mumumwp aflcouno mo ommmm one .mm wands

30

Figure 5. The effect of chronic dietary administration of
fluridone to male Mallards upon mean percent change
of initial body weight.

31

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33

Figure 6. The effect of chronic dietary administration of
fluridone to female Mallards upon mean percent
change of initial body weight.

34

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35

Egg production data (egg/hen/day) for treatment groups
display no significant differences from their respective
control values. These data are shown in Table 7. Mean egg
production plotted against time (weeks) is presented in
Figure 7.

Data from all sets per treatment group were combined for
analysis of the reproductive parameters. No significant
differences were noted for percent set, percent fertile,
percent live day 14, percent live day 21, percent live prem
pip, percent pipped, percent hatched, or percent survived.

In the percent pipped live category, 100 and 300 ppm treat-
ment groups were shown to be significantly lower than the con-
trol values (Table 8). Mallard reproductive parameters were
converted to a percent of the number of eggs laid and are
displayed as a histogram in Figure 8 (Figures in Table 9).

Analysis of the eggshell measurements failed to show
a significant difference between treatment groups and the
control (Table 8).

The pathology findings provided by Eli Lilly and Co.
are presented in Table 10. Gross and microscopic examinations
did not reveal any fluridone-related pathologic alterations.

Bobwhites: Chronic ingestion of dietary fluridone did

 

not result in greater mortality for either female or male
Bobwhites. Most deaths were caused by indeterminable factors
(Table 11).

Feed consumption of the birds on treated diets was not

significantly different from the controls as shown in Table 12.

36

 

 

 

 

 

Table The effect of chronic dietary administration of
fluridone to Mallards upon egg production (eggs/
hen/day : S.D.)
Set Dietary treatment (ppm)
number 0 100 300 1000
1 0.4931 0.6002 0.643 0.429
(10.0943) (:0.1598)a (:0.0824)a (i0.l4l8)a
2 0.618 0.643 0.586 0.500
(:0.0888) (:0.l409)a (:0.0855)a (:0.0677)a
3 0.666 0.564 0.575 0.693
(10.1079) (:0.3000)a (:0'1395)a (:0.1638)a
4 0.549 0.513 0.475 0.500
(:0.1365) (:0.1483)a (i0.0883)a (:O.1268)a
5 0.447 0.323 0.294 0.443
(:0.1876) (:0.0980)a (:0'0935)a (:0'1201)a
6 0.278 0.395 0.234 0.393
(:0.0553) (:0.0518)a (:0.1003)a (:0.1366)a
7 0.319 0.340 0.217 0.307
(10.1719) (110.0984)a (:0.2103)a (:0'1655)a
8 0.207 0.207 0.179 0.109
(i0.0706) (10.1885)a (:0'2040)a (:0'0870)a
9 0.244 0.260 0.241 0.054
(:0.0930) (:0'1077)a (i0'1897)a (:0'1070)a
10 0.101 0.050 0.116 0.009
(10.1554) (: 0.0588)a (10.1683)a (:0‘1800)a
l n = 4 for all means.
2 Means with subscript "a" are not significantly different

from control values (P > 0.05).

37

Figure 7. The effect of chronic dietary administration of
fluridone to Mallards upon mean egg production.

38

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39

Table 8 . The effect of chronic dietary administration of
fluridone to Mallards on mean reproductive
parameters (: S.D.) for all sets combined.

 

 

Dietary concentration (ppm)

 

 

Reproductive
Parameter o 100 300 1000
% set 80.21 84.0 2 85.5 84.9
+4.73 +2.80a +2.50a :3’97a
% fertile 93.2 79.2 86.2 84.9
+3.69 +ll.38 +10.95 +17.19
- - a — a — a
% live 98.3 96.9 96.3 98.1
day 14 10.61 :0.613a i2’17a ‘:2.78a
% live 99.1 98.6 96.8 98.0
day 21 +0.600 +1.81a i2‘23a +1.86a
% live 80.7 80.4 88.7 85.5
prepip +9.78 +10.81 +3.59 +1.80
—4 - a - a -» a
% pipped 99.7 99.3 98.8 100.0
+0.600 +1.35 +0.826 +0
— —- a —- a .— a
% pipped 97.5 94.6 95.9 94.2
live +1.20 +0.789 +1.59 +1.86
— — b — a —. b
% hatched 96.0 92.4 94.5 97.7
+1.67 +2.01 +3.42 +1.68
-— <— a — a — a
% survived 94.7 98.8 98.6 95.5
+2.21 :1'62a :l.94a :4'37a
Eggshell 0.429 0.430 0.424 0.420
thickness(mm) 19.0121 i0.0208a 10.0248a 10.0171a

 

1'Reproductive parameters (except eggshell thickness) expressed
as a percentage of the previous parameter.

Means with the subscript "a" are not significantly different
from their respective controls (P > 0.05); means with the
subscript "b" are significantly different from their
respective controls (P < 0.05).

40

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Mallard reproduction parameters expressed in a conintuum as a percent
of the number of eggs laid.

Figure 8.

41

Table 9 . Mallard reproductive parameters expressed as a
percentage of eggs laid.

 

 

Dietary concentration (ppm)

 

 

Parameter

0 100 300 1000
% fertile 74.7 66.5 73.7 72.1
% live 73.4 64.4 71.0 70.7
day 14
% live 72.7 63.5 68.7 69.3
day 21
% live 58.7 51.1 60.9 59.5
prepip
% pipped 58.5 50.7 60.2 59.5
% pipped 57.0 50.0 57.2 56.0
live
% hatched 54.7 46.2 54.5 54.7
% survived 51.8 45.6 53.6 52.2

 

42

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43

 

 

 

Table 11. Interim deaths for Bobwhites fed fluridone.
Dietary Cage Bird Date
level # # Sex of death Cause of death
0.00% 16 8553 F October 12 Unknown
0.01% 29 8671 M January 5 Unknown
0.10% 3 8580 M January 18 Starvation
0.03% 57 8574 F January 18 Myocardial degen-
eration & myositis
of skeletal muscle
0.10% 56 8505 M January 24 Unknown

 

44

 

 

 

 

Table 12 . The effect of chronic dietary administration of fluri-
done to Bobwhites upon food consumption (grams/hird/
day : S.D.).
Weeks on Dietary treatment (ppm)
treatment 0 100 300 1000
2 16.82 16.8 17.0 17.7
(i 1.14) (: 1.41?a (: 1.59)a (: 1.83)a
4 17.7 16.5 17.8 18.6
(i 1.89) (: 1.85)a (: 1.70)a (: 2.14)a
6 18.7 18.3 18.9 19.0
(i 1.55) (i 1.53)a (i 1.56)a (i 1.48)a
8 19.0 18.6 18.9 19.4
(i 1.35) (: 1.00)a (: 1.28)a (: 1.74)a
10 19.9 18.6 20.5 20.2
12 21.8 21.4 23.6 22.2
(i 2.40) (: 1.81)a (: 2.82)a (: 2.63)a
14 23.7 23.3 24.5 23.8
(i 2.78) (i 1.91)a (: 2.21)a (: 2.55)a
16 24.2 25.1 24.8 25.6
(i 2.65) (: 2.66)a (i 3.60)a (: 3.81)a
18 26.0 26.0 27.3 27.3
(i 3.17) (: 2.44)a (: 3.81)a (: 4.16)a
20 27.6 27.3 26.5 26.7
(i 4.67) (i 3.52)a (i 3.94)a (: 5.00)a
22 30.5 28.8 29.1 29.8
(i 5.75) (: 3.74)a (: 5.36)a (: 5.64)a
24 28.8 30.6 27.6 28.1
(i 8.27) (_+_10.o3)a (i 5.17)a (: 5.52)a

 

1Food consumption values represent a biweekly average.

2n = 15 per treatment for all weeks.

3Means with subscript "a" are not significantly different
from their respective controls (P > 0.05).

3;;

ti)

fic

OD

r8pz
Perc
nifi
trea
trol
Table
enCes
the 5
five
ingub
the a:

reprOC

45

A graphic representation of these data is presented in
Figure 9.

Neighter male nor female Bobwhites fed the treated
diets exhibited significant body weight differences (expres-
sed as a percent change of initial body weight) from the
controls (Tables 13 and 14). Figures 10 and 11 display
these changes over time for the males and females, respec-
tively.

Analysis of the egg production data revealed no signi—
ficant differences between the control birds and the birds
on treated diets except for the 100 ppm treatment group
during the fourth set. Egg production for the 100 ppm
group was significantly higher than the control group
(Table 15). These data are presented graphically in Figure 12.

Significant differences were present in two of the
reproductive parameters (Table 16). Percent fertility and
percent hatchability of the 100 ppm treatment group was sig-
nificantly greater than the control group. The 300 ppm
treatment group was also significantly greater than the con-
trol for percent fertility. Note the discrepancies in
Table 16 where notated with astericks. These value differ-
ences are due to the elimination of sets one and five when
the data Were no longer available. Loss of data from set
five was due to the accidental lack of egg transfer from
incubator to hatcher. Loss of data from set one was due to
the accidental absence of parent stock information. The

reproductive parameters were converted to a percent of the

46

Figure 9. The effect of chronic dietary administration of
fluridone to Bobwhites upon mean food consumption.

47

V“

«N ON

up

b2m1h(u¢.— ZO m¥uu3
or 3. up Or a o

 

.5... coop o
8.03 CON I
Ea.— OO—. 0

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On

on

(PM/6) nouawnsuoa coo.) uvaw

 

weeks
"eatme

 

10

24

‘ BCdy WI
b9fore

n :15
1000p

from t)

48

 

 

 

 

 

Table 13. The effect of chronic dietary administration of
fluridone to malelBobwhites upon mean percent
change of initial body weight (: S.D.
Weeks on Dietary Concentration (Ppm)
treatment 0 100 300 1000
2 3
0 103.7 105.4 104.4 107.0
(: 4.84) (: 6.51)a (: 4.32)a (: 6.27)a
2 106.5 108.9 106.6 109.4
(: 5.14) (: 5.82)a (: 4.57)a (: 4.22)a
4 111.0 113.1 111.0 113.9
(3'. 4.07) (i 5'86)a (i 5.43)al (i 5.23)a
8 113.9 116.8 114.8 117.6
(1 5.35) (: 6.34)a (: 6.08)a (: 6.51)a
10 112.7 117.0 113.9 117.4
(3: 5.08) (: 5.62)a (i 5-92)a (: 6.10)a
24 115.8 117.0 113.0 117.8
(: 9.70) (ill-15):.) (: 7.50)al (i 7.66)a
1

before week 0).

2

1000 ppm (n = 13) for week 24.

3

n = 15 for all means on all dates, except 100 ppm (n

(P > 0.05).

Body weight on 9—5-79 taken as initial body weight (2 weeks

= 14) and

Means with the subscript "a" are not significantly different
from their respective control values

49

Figure 10. The effect of chronic dietary administration of
fluridone to Bobwhite males upon mean percent
change of initial body weight.

50

plug—(ma... 20 nub—um;

m w

Eng 000' 0
Eng 00m -
Eon cow 0

405.200...

1..)
1:,
F

.0:

5g

romp

.0:

 

1H9|3M A008 1Vl1lNl :lO ‘7. NVEW

51

Table ‘14. The effect of chronic dietary administration of fluri-
done to female Bobwhites upon mean percent change of
initiallbody weight (15.0.)

 

 

 

 

Weeks on Dietary concentration (ppm)
treatment 0 100 300 1000
0 103.92 106.6 107.5 106.1
(: 5.30) (1 4.39); (i 4.74)a (: 3.94)a
2 p109.9 107.6 107.9 108.0
(: 5.43) (: 7.07)a (: 4.58)a (: 3.57)a
4 112.6 111.6 114.4 112.1
(1 6.79) (: 5.86)a (: 5.93)a (: 3.92)a
6 117.1 116.4 119.0 118.0
(1 5.97) (: 4.87)a (: 7.02)a (: 4.36)a
8 119.9 118.9 119.1 121.5
(1 9.23) (: 7.57)a (: 9.66)a (: 9.66)a
24 128.9 132.2 128.9 129.8
(112.87) (: 6.07)a (i13-75)a (: 8.11)a

 

1 Body weight on 9-5-79 taken as initial body weight.

2 n = 15 except controls (n = 14) after 10-3-79 and 300 ppm (n =
14) on 3-5-80.

3 Means with the subscript "a" are not significantly different

from their respective control values (P > 0.05).

52

Figure 11. {The effect of chronic dietary administration of
fluridone to Bobwhite females upon mean percent
change of initial body weight.

53

0

Flu Ip<w~= 20 33w?
t N

 

82.82 6
Bacon ..
EAQOOP 0

405.200 1..

oo—

o..—

On—

on—

ov—

lH9l3M A008 1VlllNl :IO "I. NVSW

54

Table 15 . The effect of chronic dietary administration of
fluridone to Bobwhites upon egg production (eggs/
female/day : S.D.)

 

 

 

 

Set Dietary concentration (ppm)
number 0 100 300 1000
1 0.4081 0.705 0.600 0.543
(: 0.3995) (: 0.3028?a (i 0.2815)a (: 0.3464)a
2 0.408 0.724 0.600 0.571
(: 0.4224) (: 0.2442)a (: 0.3110)a (: 0.3327)a
3 0.418 0.743 0.629 0.562
(: 0.4247) (: 0.2034)a (i 0.3138)a (: 0.3214)a
4 0.418 0.771 0.524 0.552
(+ 0.4018) (+ 0.2006), (+ 0.3084) (+ 0.3884),
— - — a — a
5 0.408 0.666 0.592 0.571
(: 0.4334) (1 0.1762)a (: 0.3060)a (i 0.2906)a
6 0.439 0.695 0.520 0.657
(1 0.4320) (: 0.1779)a (: 0.3995)a (: 0.3650)a
7 0.337 0.581 0.306 0.467
(i 0.3783) (i 0.2441)a (: 0.3307)a (: 0.3718)a
8 0.337 0.457 0.276 0.333
(: 0.3945) (: 0.2655)a (: 0.314)a (: 0.3399)a
9 0.357 0.514 0.306 0.362
(t 0.3531) (i 0.2741)a (i 0.3536)a (i 0.3450)a
10 0.388 0.533 0.316 0.381
(: 0.3605) (i 0.2723)a (i 0.3322)a (: 0.3222)a
11 0.398 0.543 0.388 0.362
(: 0.3848) (: 0.2966)a (i 0.3038)a (i 0.2797)a

 

1 n = 15 except controls (n=14 for all sets) and 300 ppm (n=14

from set 5 through set 11).

Means with the subscript "a" are not significantly different
from their respective control values (P > 0.05); means with the
subscript "b" are significantly different from their respective
control values (P < 0.05).

Figure 12. The effect of chronic dietary administration of
fluridone to Bobwhites upon mean egg production.

 

 

56

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57

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 16. The effect of chronic dietary administration of-
fluridone to Bobwhites on mean reproductive
parameters for all sets combined.

conc. - - -
set laid set fert11e set fertile

1229)

0 343 424 80.9 225 343 65.6
100 627 724 86.6: 470 627 75.0a
300 410 512 80.1a 348 410 84°9b

1000 467 563 82.9a 318 467 68.1a
# live # % live # live # live % live
ppm day 11 fertile day 11 day 18 day 11 day 18

0 215 225 95.6 214 215 99.5
100 461 470 98.1a 453 461 98.3a
300 334 348 96.0a 324 334 97.0a

1000 291 318 92.1a 287 293 98.0a
ppm # live # live % 2 # # live %
prepip day 18 prepip pipped prepip pipped

0 177 187 94.7 173 177 97.7
100 394 406 97.0a 387 394 98.2a
300 286 293 97.6a 280 286 97.6a

1000 240 261 92.0a 234 240 97.5a

 

 

 

cont'd

58

 

 

 

 

 

 

 

 

 

Table ‘15 (con't).
ppm # pipped. # % pipped #pipped %
live pipped live hatched live hatched
0 171 173 98.8 75 171 43.9
100 382 387 98.7a 261 382 68.3b
300 279 280 99.6a 150 279 53.8a
1000 227 234 97.0a 126 227 55.5a
ppm 6 # 5 Eggshell thickness
‘ surv1ved hatched surv1ved ppm (mm)
0 48 64 75.0 0 0.249 _ 0.0096
100 174 233 74.7a 100 0.243 : 0.0183
300 95 129 73.6a 300 0.249 : 0.0171
1000 85 111 76.6a 1000 0.248 : 0.0144

 

 

 

 

Percentages with the subscript "a" are not significantly
different (P.> 0.05) from their respective control values;

percentages with the subscript "b" are significantly different

(P < 0.05) from their respective control values.

VM

From this point on, one set (set #5) was eliminated; number

live day 18 values will not coincide with values in the

previous block.

From this point on, an additional set (set #1) was eliminated;

number hatched values will not coincide with values in the

previous block.

59

due to the accidental absence of parent stock information.
The reproductive parameters were converted to a percent of
the number of eggs laid and are displayed as a histogram in
Figure 13 and are liSted in Table 17.

The pathology findings provided by Eli Lilly and Co. are
listed in Table 18. Gross and microscopic examination pre-

sented no compound-related pathological alterations.

60

Table 17. Bobwhite reproductive parameters expressed as a
percentage of eggs laid.

 

 

Dietary concentration (PPm)

 

 

Parameter

0 100 300 1000
% set 80.9 86.1 82.9 80.1
% fertile 53.3 64.6 70.4 54.4
% live 50.7 62.8 67.8 49.8
day 11
% live 50.4 62.2 67.4 48.8
day 18
% live 47.7 60.3 65.8 44.9
Prepip
% pipped 46.1 59.2 64.2 43.8
% pipped 45.5 58.6 63.8 42.5
live
% hatched 20.0 39.9 34.2 23.6
% survived 15.0 29.8 25.2 18.1

 

61

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64
DISCUSSION

Shortly after the introduction of DDT in the 1940's
the toxic effects of pesticides on avian wildlife became
well-documented. Although the organochlorine compounds
have caused most of the pesticide-related toxicity problems,
other pesticides have been incriminated as well (McEwen
and Stephenson, 1979). Some of these adverse effects in-
clude increased mortality (Fergin and Schafer, 1977;
Wallace, 1959; Pearce and Peakall, 1977; Turtle et al., 1963;
Hunt, 1960; Ratcliffe, 1969), decreased food consumption
(Heinz, 1979; Davison and Sell, 1974; Ernst and Ringer,
1968; Keith and Mulla, 1966; Linduska and Springer, 1957),
decreased body weight (Nusz et al., 1976), increased organ
weights (Strik et al., 1980), changes in the levels of cer-
tain enzymes or blood parameters (Iturri et al., 1978;
Meydani and Post, 1979; White, 1976), and sublethal repro-
ductive problems (Blus et al., 1979; Faber et al., 1972;
Faber and Hickey, 1973; Frank et al., 1975; Odsjo and
Sondell, 1976; Peakall, 1970; Jefferies, 1957; and Heinz,
1974). Lethality was dramatic and easily observed while
the sublethal reproductive problems were more subtle thereby
taking longer to document and understand. Population
declines attributed to reproduction problems have been
characterized by reduced clutch size, egg breakage, high
embryo death rates, high chick mortality rates, and unusual

nesting behavior (McEwen and Stephenson, 1979). The most

65

documented and researched effect is that of eggshell thinning
(Ratcliffe, 2967; Hickey and Anderson, 1968; Cooke, 1973;
Davison and Sell, 1974).

In an effort to predict and control these pesticide-
related effects on avian species, the U.S. Environmental
Protection Agency (EPA) has proposed the requirement of
avian toxicity studies for partial fulfillment of pesticide
registration (Federal Register, 1978). These studies, along
with other data provided by the manufacturer, enable EPA to
evaluate the hazards posed to birds as the result of a par-
ticular pesticide‘s use (Federal Register, 1978). The pro-
posed rules require both an avian single—dose oral LDSO and
an avian dietary LCSO to support the registration of all
manufacturing-use products and all formulated products
intended for outdoor application. A subsequent avian repro-
duction study would be required if any of the following con-
ditions exist: 1) the pesticide is persistent in the en-
vironment to the extent that toxic amounts on avian feed
could be expected under normal use; 2) the pesticide is
stored or accumulated in plant or animal tissues; 3) the
pesticide is intended for use under conditions where birds
may be subjected to repeated or continued exposure to the
pesticide especially during the breeding season; or 4) any
exisiting test information indicates that reproduction may
be adversely affected by the use of the pesticide (Federal
Register, 1978).

Acute oral toxicity tests measure the inherent toxicity

of a
test
sens
meth
of t
more
duri
leve
LDSO
leth

(Fed.

ures
test
from
Chang
Data
Wild]
test
field
bilit
Via 3
total
three
as th
dry d

Parts

66

of a chemical (Tucker and Crabtree, 1970). Data that such
tests provide are used mainly for comparisons of species'
sensitivity or a pesticide's relative toxicity. This
method involves the administration (by peroral intubation)
of the test chemical as a single dose to groups of five or
more birds. The birds are observed for fourteen days
during which time mortality is recorded. Sufficient dosage
levels are used to statistically calculate an LD50. The
LDSO is defined as the dosage of test chemical that is
lethal to fifty percent of the experimental population
(Federal Register, 1978).

The avian dietary LC50 is a subacute test which meas-
ures a species' reSponse to a diet contaminated with the
test chemical (Heinz, 1979). Subacute studies are different
from acute studies in the sense that they allow for metabolic
changes that occur more readily in repeated dietary exposure.
Data provided by subacute studies are often more useful in
wildlife toxicology assessments than acute data since the
test chemical is consumed in a manner more similar to actual
field situations. The speed of incapacitation and reversi-
bility of compound-related effects can also be determined
via subacute studies. The avian dietary LCSO is run for a
total of eight days - five days on treated feed followed by
three days on uncontaminated feed. Toxicity is expressed
as the median lethal concentration (LC50) of a chemical in
dry diet. The concentration is most commonly presented as

parts per million (ppm) in feed (Federal Register, 1978).

it
ir.
ac
an

pr

tal

is

am

67

The present study is an example of the third type of
prOposed avian study and adheres to the guidelines prOposed
by EPA (Federal Register, 1978). Avian reproduction studies
determine a pesticide's effect on all aspects of reproduc-
tion. More detailed information can be obtained from the
Materials and Methods section of this thesis.

Other conditional avian studies have been proposed and
would be requested on a case-by-case basis. These include
a short-term (small pen) field test, a long-term (large)
pen field test, and a full scale field test which would
evaluate hazard to wildlife, including birds. These
studies provide exposure through pesticide application as
would occur in the field situation.

Toxicity of Fluridone.

 

Since chemicals vary in their ability to produce lethal-
ity, dose categories have been devised which rate a compound
in terms of its toxicity. Classification systems exist for
acute oral LD50's, dermal LD50's, aquatic 96-hour LCSO's,
and subacute dietary LCSO's. These rating systems are
presented in Tables 19 and 20.

With the information provided in the above mentioned
tables and fluridone toxicity data from Tables 1 and 2, it
is obvious that fluridone was not very toxic to the mammalian
and avian species tested. However, it was moderately to
highly toxic to fish.

Herbicides are generally less toxic to animals than

insecticides. This makes sense when consideration is given

68

.Hmum3 mo umuwa Mom Hmoflamno m0 m5

 

 

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.ommH .mmOHDOmmm Hohsumz mo acmsunmmmo cmmflsoflz H

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H

69

Table 20. Classification system used to rate or compare
the subacute dietary LCSOS of chemicals .

 

 

 

 

Rating Subacute dietary LC50 (ppm)
Highly toxic <40

Very toxic 41-200

Moderately toxic 201-1,000

Slightly toxic l,001-5,000

Probably not toxic >5,000

1

Heinz, 1979.

70

to the fact that plants differ from animals in major aspects
of their morphology and physiology (Doull et al., 1980).

The lower toxicity of herbicides over insecticides is
demonstrated in Table 21 which lists the LC50s of several
herbicides and insecticides administered to Bobwhites. The
LC50 of fluridone to Bobwhites is:> 5,000 ppm and therefore,
is relatively non-toxic (Table 1). The LDO (highest non-
lethal dose) of fluridone to Mallards is 21,000 mg/kg

(Table 1). A list of the acute oral LDSOS of several other
pesticides administered to Mallards is presented in Table 22.
Fluridone is less toxic than most of the other pesticides
listed.

LCSOS and LDSOS provide minimal information when evalu—
ating a compound's potential hazard to avian life. Chronic
studies can provide valuable information for use in calcula-
ting the maximum daily dietary intake of a compound tolerated
without adverse effects (Kenaga, 1973). Comparison of the
level of pesticide tolerated by birds in toxicity studies
with the level expected in field situations provides a
basis for assessing a pesticide's hazard to birds.

The approximate daily intake of fluridone for Mallards
and Bobwhites during the course of the present study is
reported in Tables 23 and 24, respectively. Prior to calcu-
1ation, the fluridone intake was expected to be greater for
Bobwhites than for Mallards. This assumption was based on
the fact that smaller sized animals consume more food on a

percent body weight basis due to the increased surface area

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71

 

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ooo.mA mcflumefim hmH coflnuouuflcom
mnm.ma xw>aflm vH Gauccm
ooo.mA uoHrommoum hm caucamfla
ooo.mA EmHoHoflm mam mo>HoHnofio
ooo.mA cousaoz mvN :OCHNMHQ
omh.a conned Ham eon
~mm.~ umnvfia Hmm mcmcuoHso
ooo.mA ouv.~ ooo.~A Haumnuao
ooo.mA mcaumuum pm canvam
lemme omoq moaoanumm lemme omen moaoauommaH

 

 

.Hmwufinanom ou

woumumficflaom moofiOHnuma can mmcflowuommcw cwouumo mo momuq mumumflo musomnsm .Hm manna

72

.onmu .mmuunouo can meuse

H

 

 

nonnaow .08 mum h.Oh camcmmxoa

moam8 a
ooo.~A e-m.c.~ «mamsmu .03 o a.~ acacsusuum
mmHaEMH a umauawu .05 m msH.NA .Hsuunuao. ca>wm
umaaa .oa mum mmo.~A o-v.~ mwflusmm .03 «um ooo.~A mnocouom
ooo.~A aafiausauaua muaaaou .05 m mo.m aooaeaammoza
moame .08 «um ooo.~A nobawm m0H08 .08 «In ma.~ scanumumm
«Hm uoasoadoum mmama .os sum oo¢.~A xmuaz
mwams .02 «um ooo.~A aauoaoam amass .05 «In ooc.~A uoHnomxonumz
mmame .oa hum n.- Hoummam mmamfiau .os cum mmv.H consumamz
amass .os eum ooo.~A nausea mmama .os cum ooo.~A mauvcau
moans .08 vim com unswwo «mama .08 m ooo.~A acanomummm
Mano nwmocwo m0H08 .08 cum mma peanuao
ooo.~A HacwnoHnoao moaosmu .os mauoa vm.m cauocm
ooo.~A oaoacoao uoaaamu .05 5-6 Ham caucamaa
um~080u .08 vim ooo.~A couoxOHOHnU mouu80u .08 «In vm.m cocwnuaa
uoflusuu .05 n ooo.~A cououco nmausmu .03 m ov~.~A eon
nouoauu .oa vun ooo.~A undue augmeuu .oa mnv com.” accouoaso
ufldflfifiu .08 m cooamA flaunuflunflfl umfigflw .06 wlv afnd Gomkflm
uudu8 .08 vim ooo.~A odouuduu08fi8< nausea“ .08 win cum canvad
acuoaanz no .mx\msc snag ocaoanuum nouuauuz no .mx\ma.omoq ocaoauuouaH

now u and

80¢ a om‘

 

 

H

.ucuuadd: 0v aaaouo wououuwcuaua uukuwuoousw can noodownuun cdouuoo m0 .mx\m=. nomad 0u=0¢. .NN wands

 

 

 

 

 

 

 

Table 23. Amount of fluridone (mg/kg/day) ingested by
Mallards.
Weeks Dietary Concentration (ppm)
on
Feed 100 300 1,000
2 12.2 34 118.5
4 12.6 39.9 121.3
6 12.8 42.6 119.1
8 11.8 31.2 115.4
18 19.8 53.4 158.4
28 15.9 32.7 146.9
Table 24. Amount of fluridone (mg/kg/day) ingested by
Bobwhites.
weeks Dietary Concentration (PPm)
on
Feed 100 300 1,000
2 8.8 26.2 90.3
4 8.2 26.1 91.3
6 8.8 27.0 89.4
8 9.2 27.1 89.3
16 11.8 35.4 116.2
24 13.2 38.1 126.5

 

74

to body weight ratio (Wilson, 1972; Prosser, 1973). One
possible explanation for the results is the Mallard feed
wastage. Pen-reared birds such as ducks and pheasants
waste a considerable amount of feed (Kenaga, 1973).
Empirical observations indicated that feed wastage was con-

siderable for the Mallards in this study. Since no mathe-

matical compensations were made during food consumption cal
culations, the Mallard food consumption data may be errone-
ously high. Feed wastage by the Bobwhites was minimal.

The highest daily fluridone intake by Mallards was
158.4 mg/kg while for Bobwhites the highest value was 126.5
mg/kg (Tables 23 and 24). Both values were calculated from
the 1,000 ppm treatment groups whose values were not signi-
ficantly lower than the control groups in any of the analyzed
parameters. This information can now be compared to the
quantity of residues expected in the environment, providing
important data necessary to evaluate this pesticide's hazard
to birds. As an example, the Mallard's daily intake of
fluridone can be used to make this comparison especially
since the reported residue data pertains to aquatic use.

Eli Lilly and Co. has established recommended fluridone
application rates for use in research trials (Lilly, 1981).
This information, plus data from a study by West et a1.
(1979) was used to approximate and evaluate a daily intake
level for Mallards presuming they consumed a quantity of
plant material equal to the quantity of diet consumed during

the reproduction study. This is only a rough approximation

75

since many variables determine food consumption and actual
residue intake. The approximation assumes similar caloric
content of both foods (plant material and prepared feed).

The application rate on one of the ponds utilized in
the study by West et a1. (1979) was 2.7 kg/ha. This rate is
higher than the 0.55—l.1 kg/ha rate suggested by Lilly for
a pond of equivalent size (Lilly, 1981). Residue analysis
calculated 3.98 ppm of fluridone residue on aquatic plants
three days after treatment. This was the highest value
reported in the study. If a Mallard were to consume 155
grams of plant material (average food consumption during
the study) when residue levels were 3.98 ppm, it's daily
intake would be 54.9 mg/kg/day. This figure is based on
an average body weight of 1123.2 grams (calculated from
body weight data obtained from this study). Even though
this hypothetical situation was maximized in terms of
application rate and dietary habits, a daily intake lower
than values estimated during this study was calculated.
This estimation supports the argument that fluridone was
administered in this study at doses high enough to be of
value in extrapolating to a real field situation. The
estimation also suggests that fluridone, when used as
recommended, should not present a hazard to Mallards.
Examination of the Study's Results.

Mallard mortality which occurred during the study was
high but was not caused by fluridone administration. The

Mallard drakes that died on test started to die shortly

76

after induction of the study's production phase. All six
drakes were dead before the first female death was reported.
A similar trend was shown by Breslin (1981) and Jones (1977).
All of the male deaths and most of the female deaths were
caused by aggressive behavior which developed in response

to the increased photoperiod (Table 3). Increasing day-
length stimulates hormone production which leads to sexual
maturation and competitive behavior (Welty, 1979; Sturkie,
1976).

Bobwhite mortality was less frequent with no deaths
attributed to aggressive behavior. Similar mortality data
were reported by Howell (1981) and Breslin (1981) whose
studies also included one pair per cage.

Body weight changes for both Mallards and Bobwhites
(Tables 3, 4, 11, and 12 and Figures 5, 6, 10, and 11) and
feed consumption data for Bobwhites (Table 10 and Figure 9)
were comparable to figures and trends reported in the
literature (Howell, 1981; Jones, 1977; Breslin, 1981;

Scott et al., 1976). Feed consumption for Mallards (Table

2 and Figure 4) was comparable to one study (Breslin, 1981),
slightly higher than another (Jones, 1977), and considerably
higher than a third (Davison and Sell, 1974).

Mallard egg production data and percentages for other
reproductive parameters were quite high (Tables 5 and 6 and
Figures 7 and 8) and in agreement with other results reported
in the literature (Breslin, 1981; Jones, 1977; Nestler et

al., 1944; Federal Register, 1978; Heath et al., 1969;

77

Davison and Sell, 1974).

Although significant in only one case (P < 0.05), the
Bobwhite egg production data were lowest for the control
group (Table 13 and Figure 12). However, egg production
values reported for all treatment groups were within the
ranges provided by other studies (Breslin, 1981; Howell,
1981; Fergin and Schafer, 1977). Fertility and hatchability
of the control group was also lower than other treatment
groups (significantly lower in two instances). These results
were unexpected and are difficult to explain since birds
and dietary concentrations were randomly assigned to the
pens and all pens were treated similarly. Although the
control group was generally less productive than the other
groups and meaningful comparisons were more difficult to
make, the lack of a dose response trend by treatment groups
reaffirms that there were no fluridone-related effects.

Some of the Bobwhite reproductive data (egg production
and embryo survival) is-comparable to data from other studies
(Wilson and Holland, 1974; Howell, 1981; Federal Register,
1978; and Breslin, 1981). Other parameters, primarily fer-
tility, hatchability, and survivability range lower or are
in the lower limits of the ranges provided by other studies
(Howell, 1981; Federal Register, 1978; Breslin, 1981;

Wilson and Holland, 1974). Figure 18 illustrates where the
most dramatic losses have occurred in the continuum of
reproductive parameters.

Bobwhite and Mallard eggshell thickness measurements

78

were both greater than those reported in the literature
(Jones, 1977; Howell, 1981; Federal Register, 1978; Heath

et al., 1969).

CONCLUSIONS

1. Chronic dietary administration of fluridone did not
affect the food consumption or body weight gain of
Bobwhites or Mallards.

2. Chronic dietary administration of fluridone did not
affect egg production or fertility of Bobwhites or
Mallards.

3. Chronic dietary administration of fluridone did not
affect embryo survival, hatchability, or survivability
(2 weeks) of the offspring (Bobwhites and Mallards).

4. Chronic dietary administration of fluridone did not

affect eggshell thickness (Bobwhites and Mallards).

APPENDICES

APPENDIX A

STRUCTURE AND SOLUBILITIES OF FLURIDONE

CHEMICAL STRUCTURE OF FLURIDONE:

 

|
cg CH3

MOLECULAR FORMULA: cwuurauo

 

SOLUBILITY:

Solvent Solubility (mg/ml)
methanol > 10.0
diethylether > 1.0
ethylacetate > 5.0
chloroform > 10.0
hexane < 0.5
dimethyl sulfoxide 240-250
tetraethyleneglycol 20-25
ethanol 30-50
dimethylformamide 400-450

79

LAYOUT OF BOBWHITE TESTING ROOM

APPENDIX B

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

*Dietary Concentration (ppm)

80

1‘1

15 300 o 16 45 o o 46

14 300 300 17 44 300 1000 47

13 100 1000 18 43 o 0 48

12 1000 300 19 42 1000 100 49

11 o 100 20 41 300 1000 so

10 o o 21 40 0 10° 51

9 100 1000 22 39 1000 300 52

8 o o 23 38 300 300 53

7 100 300 24 37 1000 300 54

6 366 1000 25 36 100 100 55

5 100 100 26 35 o 1000 56

4 6300 100 27 34 1000 300 57

3 1000 0 28 33 300 100 58

2 1000 100 29 32 100 o 59

Cige 'o* 1000 30 31 100 1000 60
, DOOR 4
I T

 

 

APPENDIX C

LAYOUT OF MALLARD TESTING ROOM

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Pen #1 Pen #12
300 ppm* 1000 ppm
Pen #2 Pen #11
1000 ppm 100 ppm
Pen #3 Pen #10
1000 ppm 0 ppm
Pen #4 Pen #9
100 ppm 300 ppm
Pen #5 Pen #8
300 ppm 1000 PP”
Pen #6 Pen #7
0 ppm 300 ppm

 

 

 

 

 

 

Door

 

Pen #13

100 ppm

 

Pen #14

0 ppm

 

Pen #15

100 ppm

 

Pen #16

OPPm

 

 

 

 

 

*
Dietary concentration of Fluridone.

81

 

 

APPENDIX D

COMPOSITION OF ADULT DUCK MASH 1

 

 

INGREDIENTS PERCENT LBS./TON
Corn, Yellow Ground 62.42 1248.4
Soybean Meal, Solvent Extracted, 19.58 391.6
Dehulled (49%)
Fish Meal 2.00 40.0
Meat and Bone Meal 4.00 80.0
Oat Groats, Rolled 2.50 50.0
Beef Tallow 0.93 18.6
Dicalcium Phosphate, Feed Grade 0.65 13.0
Limestone 6.82 136.4
Trace Mineral Premix TK-Ol 0.10 2.0
(1.02)
Salt 0.30 6.0
Vitamin Premix TK-Ol (1.03)3 0.50 10.0
Methionine Hydroxy Analog 0.20 4.0

 

100.00 2000.0

1Eli Lilly and Company.

2Trace mineral premix provides 75 mg of manganese, 50 mg of
zinc, 25 mg of iron and 1 mg of iodine per kg of complete
food.

3Vitamin premix provides 3000 IU of vitamin A, 900 ICU of
vitamin D, 40 mg of vitamin E, 0.7 mg of vitamin K, 1000 mg
Of choline, 70 mg of niacin, 4 mg of pantothenic acid, 4 mg
of riboflavin, 100 mcg of vitamin B12. 100 mcg of biotin
and 125 mg of ethoxyquin per kg of complete feed.

82

APPENDIX E

CALCULATED ANALYSIS OF ADULT DUCK MASH

 

Protein, % 18.5
Met. Energy,

KCal/Kg 2893
ME/P Ratio 156.4(71.l)
Fat, 8 3 80
Fiber, % 2 58
Ash, 8 10.97
Calcium, % 3.25
Phosphorus, % 0.66
Available Phos-

phorus, % 0.46
Manganese, mg/kg 89.0
Iron, mg/kg 105.7
Copper, mg/kg .9'7
Zinc, mg/kg 76.6

Selenium, mcg/kg 107.
Magnesium, mg/kg 2172
Potassium, mg/kg 6706

Sodium, mg/kg 1821
Iodine, mg/kg 1
Vitamin A, IU/kg 5060
Vitamin E, mg/kg 56.2

Vitamin D, ICU/kg 900

Vitamin K, mg/kg

Choline, mg/kg
Niacin, mg/kg

Pantothenic Acid,
mg/kg

Vitamin 36' mg/kg
Riboflavin, mg/kg
Thiamine, mg/kg
Folic Acid, mcg/kg
Vitamin 812, mcg/kg
Biotin, mcg/kg
Arginine, %
Lysine, %

Glycine, %
Methionine, %
Cystine, %

Total Sulfur Amino
Acids

Tryptophan, %
Linoleic Acid, %

0.7

1804
85.2
10.9

7.0
5.6
2.9
1802
107
228
1.29(6.97)
0.97(5.24)
1.10(5.94)
0.48(2.S9)
0.26(1.41)

0.74(4.00)
0.23(1.24)
1.24

 

1Eli Lilly and Company.

2A8 supplied from vitamin premix; content of other dietary
ingredients not included.

3

as a percent of dietary protein.

83

Values in parenthesis represent the amino acids expressed

APPENDIX B

COMPOSITION OF ADULT BOBWHITE MASH

1

 

 

 

 

INGREDIENTS PERCENT LBS./750 lbs.
Corn, Yellow Ground 49.37 370.28
Soybean Meal, Solvent Extracted,

Dehulled (49%) 35.67 267.52
Fish Meal, Menhaden 2.00 15.00
Corn Distillers Dried Solubles 4.00 30.00
Beef Tallow 1.00 7.50
Dicalcium Phosphate, Feed Grade 2.88 21.60
Calcium Carbonate (Limestone) 3.88 29.10
Vitamin premix TK-Ol (1.03)2 0.50 3.75
Trace Mineral premix TK-Ol (1.02)3 0.10 0.75
Salt 0.30 2.25
Methionine Hydroxy Analog 0.30 2.25

100.00 750.00

1Eli Lilly and Company.

2Vitamin premix provides 3000 IU of vitamin A, 900 ICU of
vitamin D, 40 mg of vitamin E, 0.7 mg of vitamin K, 1000 mg
Of choline, 70 mg of niacin, 4 mg of pantothenic acid, 4 mg
100 mcg of biotin and

of riboflavin, 100 mcg of vitamin B
125 mg of ethoxyquin per kg of comp ete feed.

3

I

Trace mineral premix provides 75 mg of manganese,

50 mg of

zinc, 25 mg of iron and 1 mg of iodine per kg of complete

feed.

APPENDIX I

1

CALCULATED ANALYSIS OF ADULT BOBWHITE MASH

 

Protein, %
met. energy, KCal/kg 2803.00
116.79(53.09)

ME/P Ratio

Fat, %
Fiber, %
Ash, %
Calcium, %

Phosphorus,
Avail. Phosphorus,

%

Manganese, mg/kg

Iron, mg/kg
Copper. mg/kg

Zinc, mg/kg

Selenium, mcg/kg

Magnesium, mg/kg

Potassium,

%

Sodium, mg/kg

Iodine, mg/kg

Vitamin A,

Vitamin D,

Vitamin E,

IU/kg
ICU/kg
mg/kg

%

24.00

3.32
2.68
9.80
2.30
1.00
0.72
93.0
116.0
18.0
84.0
121.0
2104.0
0.95
1626.0
1.0
4629.0
900.0
55.2

Vitamin K, mg/kg 0.7
Choline, mg/kg 2139.0
Niacin, mg/kg 97.0
Pantothenic Acid,

mg/kg 12.6
Vitamin B6’ mg/kg 7.5
Riboflavin, mg/kg 6.2
Thiamine, mg/kg 2.7
Folic Acid, mg/kg 1.6
Vitamin 312’ mcg/kg 103.0
Biotin, mcg/kg 301.0
Arginine, % 1.717(7.15)2
Lysine, % 1.380(5.75)
Glycine, % 1.248(5.20)

Methionine, % 0.629(2.62)
Cystine, % 0.353(1.47)

Total Sulfur 0.982(4.09)

Amino Acids, %
TryptOphan, % 0.337(1.40)

Linoleic Acid, % 1.22

 

1Eli Lilly

2

and Company.

percent of dietary protein.

87

Values in parenthesis represent the amino acids expressed as a

APPENDIX J

COMPOSITION OF BOBWHITE CHICK MASHl

 

 

 

 

INGREDIENT PERCENT LBS./TON
Corn, Yellow Ground 46.52 348.90
Soybean Meal, Solvent Extracted,

Dehulled (49%) 41.26 309.45
Fish Meal, Menhaden 4.00 30.00
Corn distillers Dried Solubles 4.00 30.00
Dried Whey 2.00 15.00
Dicalcium Phosphate, Feed Grade 0.39 2.92
Calcium Carbonate (Limestone) 0.73 5.48
Vitamin premix TK-Ol (1.03)2 0.50 3.75
Trace Mineral Premix TK-Ol (1.02)3 0.10 0.75
Salt 0.20 1.50
Methionine Hydroxy Analog 0.30 2.25

100.00 750.00

1Eli Lilly and Company.
2

Vitamin premix provides 3000 IU of vitamin A, 900 ICU of

vitamin D, 40 mg of vitamin E, 0.7 mg of vitamin K, 1000
mg of choline, 70 mg of niacin, 4 mg of pantothenic acid,

4 mg of riboflavin, 100 mcg of Vitamin B
biotin and 125 mg of ethoxyquin per kg 0

3

i280

Trace mineral premix provides 75 mg of manganese, 50 mg

100 mcg of
mplete feed.

of zinc, 25 mg of iron and 1 mg of iodine per kg of

complete feed.

88

APPENDIX K

1

CALCULATED ANALYSIS OF BOBWHITE CHICK MASH

 

Protien, % 27.99 Vitamin K, mg/kg 0.7
Met. Energy, KCal/kg 2861.0 Choline, mg/kg 2330.0
ME/P Ratio 102.22(46.46) Niacin, mg/kg 102.0
Fat, % 2.46 Pantothenic acid,

Fiber, % 2.79 mg/kg 14.3
Ash, 3 5.54 Vitamin 36' mg/kg 7.8
Calcium, % 0.70 Riboflavin, mg/kg 6.9
Phosphorus, % 0.65 Thiamine, mg/kg 2‘8
Avail. Phosphorus, % 0.34 Folic ACid' mg/kg 1-3
Manganese, mg/kg 95,0 Vitamin 312: mcg/kg 107-0
Iron. mg/kg 123.0 Bi°tin' ”Cg/k9 324-0 2
Copper, mg/kg 20.0 Arginine, % 1.991(7.11)
Zinc, mg/kg 89.0 Lysine, % 1.675(5.98)
Selenium, meg/kg 168.0 Glycine, % 1'488(5°32)
Magnesium, mg/kg 2146.0 Methionine, % 0.710(2.54)
Potassium, % 1.13 Cystine, % 0.413(1.48)
sodium' mg/kg 1629'0 Toiginguigigs, % 1.124(4.02)
IOdine' mg/kg 1'0 Tryptophan, % 0.169(0.60)
Vitamin A' IU/kg 4535'0 Linoleic Acid, % 1.17
Vitamin D, ICU/kg 900.0

Vitamin E, mg/kg 54.8

 

1Eli Lilly and Company.

2

as a percent of dietary protein.

89

Values in parenthesis represent the amino acids expressed

LITERATURE C ITED

9O

LITERATURE CITED

Arnold, W. R. 1979. Fluridone - a new aquatic herbicide. J.
Aquat. Plant Manage. 17:30-33.

Banks, P. A., M. L. Ketchersid, and M. G. Merkle. 1979. The
persistence of fluridone in various soils under field
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Banks, P. A. and M. G. Merkle. 1979. Soil detection and
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Bartels, P. G. and C. W. Watson. 1978. Inhibition of caro-
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Berard, D. F., Rainey, D. P., and C. C. Lin. 1978. Absorp-
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selected crop species. Weed Sci. 26(3):250-254.

Blus, L. J., T. G. Lamont, B. S. Neely, Jr. 1979. Effects
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Breslin, W. J. Personal Communication. 22 June 1981.
Cochrane, R. L. Personal Communication. 6 July 1981.

Cooke, A. S. 1973. Shell thinning in avian eggs by environ-
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Davison, K. L. and J. L. Sell. 1974. DDT thins shells of
eggs from Mallard ducks maintained on ad libitum or con-
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222-232.

Doull, J., C. D. Klaassen, and M. O. Amdur. 1980. Casarett
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Ernst, R. A. and R. K. Ringer. 1968. The effect of DDT,
Zectran, and Zytron on the packed cell volume, total
erythrocyte count, and mean corpuscular volume of
Japanese quail. Poult. Sci. 47(2):639-643:’

Faber, R. A. and J. J. Hickey. 1973. Eggshell thinning,
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Faber, R. A., R. W. Risebrough, and H. M. Pratt. 1972. Organo-
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Federal Register, Monday, July 10, 1978. Part II. Environ-
mental Protection Agency; Registration of Pesticides in
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Federal Register, Tuesday, October 28, 1980. Fluridone;
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Federal Register, Tuesday, March 17, 1981. Tolerances for
pesticides in food administered by the Environmental
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Fergin, T. J. and E. C. Schafer. 1977. Toxicity of dieldrin
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Arch. Environm. Contam. Toxicol. 6:213-219.

Frank, R., M.V.H. Holdrinet, and W. A. Ripley. 1975. Residue
of organochlorine compounds and mercury in birds' eggs
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Gill, J. L. 1978a. Design and Analysis of Experiments in the
Animal and Medical Sciences. Volume 1. Iowa State
University Press, Ames, Iowa. pp. 409.

 

 

Gill, J. L. 1978b. Designwand Analysis of Experiments in the
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State University Press, Ames, Iowa. pp. 173.

 

Heath, R. G., J. W. Spann, and J. F. Kreitzer. 1969. Marked
DDE impairment of Mallard reproduction in controlled
studies. Nature 224:47-48.

Heinz, G. 1974. Effects of low dietary levels of methyl-
mercury on Mallard reproduction. Bull. Environ. Contam.
Toxicol. 11(4):386-392.

Heinz, G. H. 1979. Methylmercury: reproductive and behavioral
effects on three generations of Mallard ducks. J. Wildl.
Manage. 43(2):394-401.

Heinz, G. H., E. F. Hill, W. H. Stickel, and L. F. Stickel.
1979. Environmental contaminant studies by the Patuxent
Wildlife Research Center. ASTM STP 693, E. E. Kenaga,
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9-35. '

Hickey, J. J. and D. W. Anderson. 1968. Chlorinated hydro-
carbons and eggshell changes in raptorial and fish-
eating birds. Science 162:271—273.

92

Howell, K. S. 1981. Toxicity of diisopropy1methy1phosphonate
and dicyclopentadiene to Bobwhite quail (Colinus
virginianus). M.S. Thesis, Michigan State University.

 

Hunt, L. B. 1960. Songbird breeding populations in DDT-sprayed
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Iturri, S., C. Rojas, M. Bergqvist, G. Calaf, and G. M. Massa.
1978. Effects of polychlorinated biphenyls and DDT on
the hematology of White Leghorn cockerels and the
Japanese quail. Arch. Biol. Med. Exp. 11(3):R81-R82.

Jeffries, D. J. 1967. The delay in ovulation produced by
p,p'-DDT and its possible significance in the field.
Ibis 109:266-272.

Jones, R. E., Jr. 1977. Toxicity of diisopropylmethylphospho-
nate and dicyclopentadiene on the Mallard (Anas
platyrhynchos). M.S. Thesis, Michigan State University.

 

Keith, J. 0., and M. S. Mulla. 1966. Relative toxicity of
five organophosphorous mosquito larvicides to Mallard
ducks. J. Wildl. Manage. 30(3):553-563.

Kenaga, E. E. 1973. Factors to be considered in the evaluation
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