Ethyl [2-(4-phenoxyphenoxy)ethyl]carbamate

Toxicity to birds

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to birds: acute oral toxicity test (LD50-only)

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-1 (Avian Acute Oral Toxicity Test)

GLP compliance:

yes

Dose method:

gavage

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

1 d

Dose descriptor:

LD50

Remarks:

single dose

Effect level:

> 7 000 mg/kg bw/day

Conc. / dose based on:

test mat.

Basis for effect:

mortality

No birds died in the control group. One bird died at 3584 mg/kg. Birds were observed to be quiet at all dose levels during the first week. Body weight gain was slightly reduced at all dose levels during days 1-3. Food consumption was not affected and no abnormalities were noted at necropsy.

Table 1. Summary of acute oral toxicity to birds.

Species	Acute oral LD50 (mg/kg bw)
Bobwhite quail Colinus virginianus	>7000

Validity criteria fulfilled:

yes

Conclusions:

In an acute oral toxitity study in birds performed in accordance with EPA 71-1, the acute oral LD50 for bobwhite quail is >7000 mg/kg bw.

Executive summary:

In this study on the acute oral toxicity of the test substance to young adult bobwhite quail (Colinus virginianus) performed following the EPA 71-1 guidance and in compliance with GLP, five male and five females per dose level (fasted overnight) were orally dosed with the test substance at dose levels of 0, 2867, 3584, 4480, 5600 and 7000 mg/kg bw, followed by a fourteen day observation period. No birds died in the control group. One bird died at 3584 mg a.s./kg. Birds were observed to be quiet at all dose levels during the first week. Body weight gain was slightly reduced at all dose levels during days 1-3. Food consumption was not affected and no abnormalities were noted at necropsy.

The acute oral LD50 for bobwhite quail is >7000 mg/kg bw.

Reference 2

Endpoint:

short-term toxicity to birds: acute oral toxicity test (LD50-only)

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-1 (Avian Acute Oral Toxicity Test)

GLP compliance:

yes

Dose method:

gavage

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

1 d

Dose descriptor:

LD50

Remarks:

single dose

Effect level:

> 3 000 mg/kg bw/day

Conc. / dose based on:

test mat.

Basis for effect:

mortality

No birds died. No treatment related effects on behaviour were observed at any dose level. Body weight gain and food consumption were not affected and no abnormalities were noted at necropsy.

Table 1. Summary of acute oral toxicity to birds

Species	Acute oral LD50 (mg/kg bw)
Mallard duck Anas platyrhynchos	>3000

Validity criteria fulfilled:

yes

Conclusions:

In an acute oral toxicity study in birds, performed in accordance with EPA 71-1, the acute oral LD50 for mallard duck is >3000 mg/kg bw.

Executive summary:

In this study on the acute oral toxicity of the test substance to young adult Mallard ducks (Anas platyrhynchos) performed following the EPA 71-1 guidance and in compliance with GLP, five male and five females per dose level (fasted overnight) were orally dosed with the test substance at dose levels of 0, 1229, 1536, 1920, 2400 and 3000 mg/kg bw, followed by a fourteen day observation period. No birds died in the control group. No birds died. No treatment related effects on behaviour were observed at any dose level. Body weight gain and food consumption were not affected and no abnormalities were noted at necropsy. The acute oral LD50 for mallard duck is >3000 mg/kg bw.

Reference 3

Endpoint:

short-term toxicity to birds: acute oral toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-2 (Avian Dietary Toxicity Test)

Qualifier:

according to guideline

Guideline:

OECD Guideline 205 (Avian Dietary Toxicity Test)

Qualifier:

according to guideline

Guideline:

other: ASTM Standard E857-87 “Standard Practice Conducting Subacute Dietary Toxicity Tests with Avian Species"

GLP compliance:

yes

Dose method:

feed

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

5 d

Dose descriptor:

LC50

Effect level:

> 5 620 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 2576 mg/kg bw

Duration (if not single dose):

5 d

Dose descriptor:

NOEC

Effect level:

1 780 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

body weight

Remarks on result:

other: equivalent to 872 mg/kg bw

One bird died in one of the control groups. No further birds died during the study. No clinical signs were noted. Body weight gain was reduced at 3160 and 5620 mg/kg diet. Feed consumption was not affected
at any dose level.

Table 1. Summary of dietary toxicity to birds

Species	Acute oral LC50 (mg a.s./kg diet)
Bobwhite quail Colinus virginianus	> 5620

Validity criteria fulfilled:

yes

Conclusions:

In a short-term dietary study in birds, performed in accordance with EPA 71-2, the 5-day dietary LC50 was >5620 mg/kg diet, equivalent to >2576 mg/kg bw/day. The 5-day NOEC was 1780 mg/kg diet, equivalent to 872 mg/kg bw/day.

Executive summary:

In this study performed following the EPA 71-2 guidance and in compliance with GLP, five groups of 10-day old Bobwhite quails of indeterminate sex (10/group) were fed the test substance in the diet at analytically confirmed concentrations of 562, 1000, 1780, 3160 and 5620 mg/kg diet, respectively, for a period of 5 days followed by a 3-day recovery period. Four control groups of ten 10-day old Bobwhite quails received feed treated with the vehicle (corn oil).

Following test initiation and continuing until termination, all birds were observed at least twice daily. A record was maintained of all mortality, signs toxicity and abnormal behaviour. Body weights by group were measured at the initiation of the test, on Day 5, and at the termination of the test on Day 8. Average estimated feed consumption was determined each test and control group for the exposure period Days 0-5, and for the post-exposure observation period, Days 6-8. Feed consumption was determined by the change in the weight of the feed presented to the birds over a given period of time.

One bird died in one of the control groups. No further birds died during the study. No clinical signs were noted. Body weight gain was reduced at 3160 and 5620 mg/kg diet. Feed consumption was not affected at any dose level.

The 5-day dietary LC50 was >5620 mg/kg diet, equivalent to >2576 mg/kg bw/day. The 5-day NOEC was 1780 mg/kg diet, equivalent to 872 mg/kg bw/day.

Reference 4

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-2 (Avian Dietary Toxicity Test)

GLP compliance:

yes

Dose method:

feed

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

5 d

Dose descriptor:

LC50

Effect level:

10 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 1356 mg/kg bw

Duration (if not single dose):

5 d

Dose descriptor:

NOEC

Effect level:

5 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 720 mg/kg bw

No birds died in the control groups. Four and nine birds died at 100 and 20000 mg/kg diet, respectively. No further birds died during the study. The mortality at 100 mg/kg diet was considered not to be dose-related, since no mortality was observed at the next two higher dose levels. No treatment related clinical signs were noted. Body weight gain and feed consumption were not affected at any dose level.

Table 1. Summary of dietary toxicity to birds

Species	Acute oral LC50 (mg/kg diet)
Bobwhite quail Colinus virginianus	10000

Validity criteria fulfilled:

yes

Conclusions:

In a short-term dietary toxicity study in birds, performed in accordance with EPA 71-2, the 5-day dietary LC50 in bobwhite quails was 10000 mg/kg diet, equivalent to 1356 mg/kg bw/day. The 5-day NOEC was 5000 mg/kg diet, equivalent to 720 mg/kg bw/day.

Executive summary:

In this study on the dietary toxicity of the test substance, performed following the EPA 71-2 guidance and in compliance with GLP, four groups of 15-day old Bobwhite quails of indeterminate sex (10/group) were fed the test substance in the diet at analytically confirmed concentrations of 100, 1000, 5000 and 20000 mg/kg diet, respectively, for a period of 5 days followed by a 6-day recovery period. Three control groups of ten 15-day old Bobwhite quails received untreated feed. No birds died in the control groups. Four and nine birds died at 100 and 20000 mg/kg diet, respectively. No further birds died during the study. The mortality at 100 mg/kg diet was considered not to be dose-related, since no mortality was observed at the next two higher dose levels. No treatment related clinical signs were noted. Body weight gain and feed consumption were not affected at any dose level.The 5-day dietary LC50 was 10000 mg/kg diet, equivalent to 1356 mg/kg bw/day. The 5-day NOEC was 5000 mg/kg diet, equivalent to 720 mg/kg bw/day.

Reference 5

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-2 (Avian Dietary Toxicity Test)

GLP compliance:

yes

Dose method:

feed

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

5 d

Dose descriptor:

LC50

Effect level:

> 20 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 5885 mg/kg bw

Duration (if not single dose):

5 d

Dose descriptor:

NOEC

Effect level:

5 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 1251 mg/kg bw

No birds died during the study. No treatment related clinical signs were noted. Body weight gain was reduced at 20000 mg/kg diet. Feed consumption was not affected at any dose level.

Table 1. Summary of dietary toxicity to birds

Species	Acute oral LC50 (mg/kg diet)
Mallard duck Anas platyrhynchos	>20000

Validity criteria fulfilled:

yes

Conclusions:

In a short-term dietary toxicity study in birds, performed in accordance with EPA 71-2, the 5-day dietary LC50 in mallard duck was >20000 mg/kg diet, equivalent to >5885 mg/kg bw/day. The 5-day NOEC was 5000 mg/kg diet, equivalent to 1251 mg/kg bw/day.

Executive summary:

In this study on the dietary toxicity of the test substance, performed following the EPA 71-2 guidance and in compliance with GLP, four groups of 13-day old Mallard ducks of indeterminate sex (10/group) were fed the test substance in the diet at analytically confirmed concentrations of 100, 1000, 5000 and 20000 mg/kg diet, respectively, for a period of 5 days followed by a 6-day recovery period. Three control groups of ten 15-day old Bobwhite quails received untreated feed. No birds died in the control groups. No birds died during the study. No treatment related clinical signs were noted. Body weight gain was reduced at 20000 mg/kg diet. Feed consumption was not affected at any dose level. The 5-day dietary LC50 was >20000 mg/kg diet, equivalent to >5885 mg/kg bw/day. The 5-day NOEC was 5000 mg/kg diet, equivalent to 1251 mg/kg bw/day.

Reference 6

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Mar 1989 to 07 Sep 1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-4 (Avian Reproduction Test)

GLP compliance:

yes

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

no

Remarks:

Acetone and corn oil

Details on preparation and analysis of diet:

DIET PREPARATION
Nominal premix preparation was as follows:
- 0 ppm: 180 mL acetone + 180 ml (162 g) corn oil + 7938.0 g ration
- 64 ppm: 12.4416 g test substance + 180 mL acetone + 180 mL corn oil + 7925.6 g ration
- 160 ppm: 31.1040 g test substance + 180 mL acetone + 180 mL corn oil + 7906.9 g ration
- 400 ppm: 77.7600 g test substance + 180 mL acetone + 180 mL corn oil + 7860.2 g ration
For the test premixes the test substance was weighed into tared containers and transferred to an approximately 600 mL beaker. The test substance solution was dissolved or suspended in acetone and corn oil with hand stirring. The ration was weighed and approximately one half was placed in the mixer. The test substance solution was added to the ration with an acetone rinse of the beaker, and mixed approximately for five minutes. The remaining ration was added to the mixer and the premix was mixed approximately for 15 additional minutes.
Each premix was then divided into four 2000 gram aliquots. Except when used immediately for final diet aliquots were placed in labeled freezer bags, reweighed and frozen.
As needed, the appropriate premix was incorporated into the final diet as follows:
- 0 ppm: 2000 g Premix + 43.60 kg ration + 2400 g limestone
- 64 ppm: 2000 g Premix + 43.60 kg ration + 2400 g limestone
- 160 ppm: 2000 g Premix + 43.60 kg ration + 2400 g limestone
- 400 ppm: 2000 g Premix + 43.60 kg ration + 2400 g limestone
The diet was mixed approximately 20 minutes in a twin shell dry blender.

HOMOGENEITY AND STABILITY OF TEST MATERIAL IN DIET
Six samples of the and each test concentration were collected on day 0 of week 1 to determine the homogeneity of the test substance in the diets. Samples to determine stability of the test compound in the diet were collected on day 7 of week 1 from feed remaining in the feeders presented to the birds. Verification samples of the control diet and each of the test diets were taken weekly after mixing. All samples were frozen after collection, stored frozen and shipped frozen on dry ice to the analytical laboratory

ANALYTICAL METHODS IN THE FEED
- Separation method: HPLC

Test organisms (species):

Anas platyrhynchos

Details on test organisms:

TEST ORGANISM
- Common name: Mallard duck
- Age at test initiation: 31 weeks
- Sexes used / mixed or single sex: mixed sex
- Disease free: yes
- Kept according to standard practices: yes
- Breeding population: The birds were approaching their first breeding season and had not been used in previous test

Limit test:

no

Total exposure duration (if not single dose):

19 wk

No. of animals per sex per dose and/or stage:

16 males and 16 females per treatment group

Control animals:

yes, concurrent vehicle

Details on test conditions:

ACCLIMATION
- Acclimation period: 8 weeks
- Feeding: The adults were fed a gamebird ration formulated for breeding birds. During the study the birds received the appropriate test or control diet from study initiation to terminal sacrifice. Water and feed were provided libitum during acclimation and during the test. All offspring received a gamebird ration formulated for young growing birds (identical to adult diet, but without the addition of limestone). The test substance was not mixed into the diet of the offspring. Feed and water were provided to the offspring libitum.
The diets contained 28 % protein minimum, 2.5 % fat minimum and 5 % fiber maximum. 5% (w/w) limestone was added to the diet to provide a calcium source.

PEN SIZE AND CONSTRUCTION MATERIALS
- Description: Adults were kept indoors in batteries of pens measuring approximately 75 X 90 X 45 cm high. The pens were constructed of galvanized wire grid and galvanized sheeting. Each pen was equipped with a feeder. Each week, sufficient feed for seven days was placed in feeders for each pen and presented to the birds. During the week additional feed was added to the feeders where excessive wastage by the birds made it necessary. Water was supplied by nipple type waterers. Hatchlings were placed in batteries of brooding pens measuring approximately 72 X 90 X 24 cm high or 62 X 92 X 25.5 cm high. The external walls and ceilings of the units were constructed of galvanized wire mesh and galvanized sheeting and floors of galvanized wire mesh. In some other units floors, walls and ceilings were constructed of vinyl-coated wire mesh.
- Caging: 1 female and 1 male per pen

NO. OF BIRDS PER STAGE OR REPLICATE
- For vehicle control: 16 males and 16 females
- For treated: 16 males and 16 females

TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 21.1°C ± 2.3°C in the adult mallard study room; Thermostats in the brooding compartment of each pen were set to maintain a temperature of 38°C approximately from the time of hatching until the birds were 5 to 7 days of age. At that time, thermostats were reset to maintain a temperature of approximately 26°C. Hatchlings were then maintained at an average ambient room temperature of 26.4°C ± 3.0°C (SD).
- Relative humidity (%): 54 ± 13
- Photoperiod: The photoperiod during acclimation and at initiation of the study was eight hours of light per day. The photoperiod was increased to seventeen hours of light per day at the beginning of week 9 and was maintained at that length until sacrifice of adult birds. The birds received approximately 130 lux of throughout the study. The photoperiod for the hatchlings was maintained by a time clock at 16 hours of light per day.

Details on examinations and observations:

MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: All adult birds were observed at least once daily throughout the study for signs of toxicity or abnormal behavior. A record was maintained of all mortalities and observations. All birds that died during the study were necropsied. As soon as practical after the death of the bird, the pen mate was sacrificed and necropsied. In addition, at the conclusion of the adult period all birds were sacrificed by cervical dislocation, necropsied and disposed of by incineration.

BODY WEIGHT
- Time schedule for examinations: Adult body weights were measured at study initiation, weeks 2, 4, 6, 8, and at terminal sacrifice. Body weights were not measured during egg laying because of the possible effects handling may have on egg production.


FOOD CONSUMPTION
- Time schedule for examinations: Feed consumption of adult mallards was measured for each pen for a seven day period every week throughout the study. Feed consumption was determined by weighing the freshly filled feeder on day 0, recording the amount any additional diet added during the week and weighing the feeder and remaining feed at the end of the seven day period. No attempt was made to quantify the amount of feed wasted by the birds, as the wasted feed is normally scattered and mixed with water and excreta. Therefore, feed consumption is presented as an estimate.

Details on reproductive parameters:

The following parameters were examined per parental pen per week:
- Eggs laid
- Eggs cracked; eggs broken
- Egg abnormalities
- Eggs set
- Eggshell thickness or eggshell strength
- Embryos viable
- Normal hatchlings
- Abnormal hatchlings
- Clinical signs of toxicity, abnormalities and mortality
- 14-day old surviving chicks
- Chick body weight at hatching and 14 days after hatching

Eggs were collected daily from all pens and marked according to the pen of origin. The eggs were then washed to prevent pathogen contamination. The eggs were stored in a cold room until incubation. At weekly intervals all eggs were removed from the cold room, counted and eggs taken for egg shell thickness measurement. The remaining eggs were candled with a egg candling lamp to detect egg shell cracks or abnormal eggs. Cracked or abnormal eggs were discarded. All eggs that were not cracked or used for egg shell thickness measurements were placed in an incubator. Eggs were candled again on day 14 of incubation to determine fertility and/or embryo viability; and on day 21 to determine embryo survival. On day 24 of incubation, the eggs were placed in a hatchery and allowed to hatch. Pedigree baskets constructed of galvanized steel wire mesh were used to keep hatchlings by pen.
All hatchlings, unhatched eggs and egg shells were removed from the hatcher on day 26 or 27 incubation. The average body of the hatchlings by pen was then determined. Hatchlings were toe and web clipped for identification by pen of origin and then housed according to the appropriate parental grouping in brooding pens until 14 days of age. The hatchlings were fed untreated diet. At 14 days of age the average body weight by parental pen of all surviving ducklings was determined. The ducklings were sacrificed with chloroform or carbon dioxide and disposed of by incineration.
Weekly throughout the egg laying one egg was collected, when available, from each of the odd numbered pens during odd numbered weeks (1,3,5, etc.) and from each of the even numbered pens during the even numbered weeks (2,4,6, etc.). The eggs were opened at the equator, the contents removed, and the shell thoroughly washed. The shells were then allowed to air dry for at least one week at room temperature. The thickness of the dried shell plus the membrane was determined measuring five points around the waist of the egg using a micrometer. Measurements were made to the nearest 0.005 mm.

Reference substance (positive control):

no

Key result

Duration (if not single dose):

19 wk

Dose descriptor:

NOEC

Effect level:

160 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

reproductive parameters

Remarks:

reduced hatchability

Remarks on result:

other: equivalent to mean measured concentration 146 mg/kg diet after correction for test material purity, equivalent to 17.7 and 18.4 mg/kg bw/day for males and females, respectively

Mortality and sub-lethal effects:

MORTALITY
There were no treatment related mortalities at any concentration tested. No mortalities occurred in the 64 ppm treatment group. A single incidental mortality occurred in the control group and in the 160 ppm and 400 ppm treatment groups.
The single mortality in the control group was a hen found dead during Week 11. No clinical sings had been noted prior to death. Necropsy revealed a bird in excellent body condition, with minor lesions of bumblefoot on the left foot. The hen was in active production with an egg in the uterus. Haemorrhagic enteritis, with injected blood vessels was evident in mid small tract. No other lesions were noted.
The single mortality in the 160 ppm treatment group was a hen found dead at the end of Week 15. No signs of toxicity were noted prior to death. Necropsy revealed a bird of moderate body weight (913 g), with lesions of bumblefoot on feet. Internally there was evidence of slight airsacculitis and lesions of extensive egg yolk peritonitis.
The single mortality in the 400 ppm treatment group was a hen found dead during Week 9. No clinical sings had been noted prior to death. The bird was in good condition (body weight - 1020 g), and no external lesions were noted. Internally the bird was unremarkable. No other mortalities occurred during the course of the study. Necropsy results of the pen mates of the birds found dead were unremarkable. Due to the nature of the lesions observed at necropsy, all mortalities were considered to be incidental to treatment.

CLINICAL SIGNS
No overt signs of toxicity were observed at any of the concentrations tested. Incidental clinical signs, such as walking on toes or observations normally associated with wear and/or interaction among pen mates, were noted at various concentrations during the course of the study. Except for the mortalities and incidental clinical signs previously noted, all birds at all concentrations appeared normal throughout the study.

BODY WEIGHT
When compared with the control group, there were no apparent treatment related effects upon body weights at any of the concentrations tested. There were no statistically significant differences between control group and the 64, 160 or 400 ppm treatment groups at any body weight interval.

FOOD CONSUMPTION
Due to excessive wastage by some birds, feed consumption was variable between pens. There were no apparent treatment related effects upon feed consumption among birds at any of the concentrations tested.

PATHOLOGY
All adults were necropsied at adult terminal sacrifice. All findings observed were considered to be incidental to treatment.

Effects on reproduction:

There were no treatment related effects upon reproductive parameters at the 64 or 160 ppm test concentrations. There was a statistically significant increase in the number of 14-day old survivors as a percentage of hatchlings at both 64 ppm (P < 0.01) and 160 ppm (P< 0.05). However, this apparent increase in offspring survivability was a function of 10 incidental mortalities in the control group due to a feeder malfunction, and was not considered to be treatment related.
There were no other statistically significant differences between the control group and the 64 or 160 ppm treatment groups for any reproductive parameter.
- Hatchability: while not statistically significant, there may have been a slight reduction in hatchability at 400 ppm. There appeared to be an increase in the number of pens with decreased hatchability (8 <50 %) when compared to the control (4 <50 %) and a decrease in the number of pens with increased hatchability (1 >80% vs. 6 >80%). There were no other apparent effects upon reproductive parameters at 400 ppm.
- Egg shell thickness: There were no apparent treatment effects upon egg shell thickness at any test concentration. When compared to the control group, there were no statistically significant differences in egg shell thickness at 64, 160 or 400 ppm.
- Offspring body weights: There was no treatment related effect upon the body weight of hatchlings or 14-old survivors at any concentration tested. When compared to the control group, there were no statistically significant differences in offspring weight at 64, 160 or 400 ppm.

Reported statistics and error estimates:

Upon completion of the study, the Dunnett‘s Multiple Comparison Procedure (4,5) was used to determine significant differences between the control group and each of the treatment groups. Sample units were the individual pens within each experimental group. Percentage data were analyzed using Dunnett‘s method following arcsine transformation. The pens in which adult occurred were not used in statistical comparisons of the reproductive data.

Validity criteria fulfilled:

yes

Conclusions:

In a reproductive toxicity study in birds performed in accordance with EPA 71-4, based on reduced hatchability at 400 mg/kg diet, the NOEC in mallard duck exposed to the test substance in the diet was 160 mg/kg diet (mean measured concentration 146 mg/kg diet after correction for purity of the test substance, equivalent to 17.7 and 18.4 mg/kg bw/day for males and females, respectively).

Executive summary:

In a reproductive toxicity study performed according to EPA 71-4 guidance and in compliance with GLP, 31-weeks-old mallard ducks were exposed to the test substance in the diet for 19 weeks. Nominal diet concentrations were 0 (control), 64, 160 and 400 mg/kg diet. There were 16 pairs per dose level, one pair per cage. The birds were maintained in a controlled environment test room. Eggs were collected daily from the onset of egg production (week 11). Hatchlings were leg banded and maintained for 14 days in cages according to pen. Reproduction endpoints were determined with eggs produced during the last 9 weeks of the study. Reproduction endpoints were: number of eggs laid, number of eggs cracked, eggshell thickness, number of fertile eggs, number of live 21-day-old embryos, number of hatchlings, hatchling weight, number of 14-day-old survivors, and chick weights at day 14 of age. For eggshell thickness determination, eggs were collected weekly throughout the egg laying period.

No treatment-related effects were observed in the adult birds. One bird died before and two birds died after onset of egg production during the exposure period of the study. The distribution of deaths among the treatment groups was one (control), one (160 mg/kg diet) and one (400 mg/kg diet). There were no treatment-related effects on body weight, body weight change, feed consumption, survival and behavioural endpoints. No substance related abnormalities were noted at necropsy.

The number of hatchlings as percentage of eggs set, fertile eggs and 21-day embryos was reduced at 400 mg/kg diet (not statistically significant), resulting in a reduction of the number of 14-day old chicks per hen and as percentage of eggs set and fertile eggs (not statistically significant). The number of 14-day old chicks as percentage of hatchlings was significantly increased at 64 and 160 mg/kg diet. This was a function of 10 incidental mortalities in the control group due to a feeder malfunction, and therefore not considered to be related to treatment. Mean body weights of hatchlings and 14-day old chicks were not affected at any dose level. 

Based on reduced hatchability at 400 mg/kg diet, the NOEC in mallard duck exposed to the test substance in the diet was 160 mg/kg diet (mean measured concentration 146 mg/kg diet after correction for purity of the test substance, equivalent to 17.7 and 18.4 mg/kg bw/day for males and females, respectively).

Reference 7

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-4 (Avian Reproduction Test)

GLP compliance:

yes

Dose method:

feed

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

21 wk

Dose descriptor:

NOEC

Remarks:

Systemic

Effect level:

400 mg/kg diet

Conc. / dose based on:

test mat.

Remarks on result:

other: equivalent to 39.2 and 35.9 mg/kg bw/day for males and females, respectively

Duration (if not single dose):

21 wk

Dose descriptor:

NOEC

Effect level:

400 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

reproductive parameters

Remarks on result:

other: equivalent to 39.2 and 35.9 mg/kg bw/day for males and females, respectively

No treatment-related effects were observed in the adult birds. One bird died before and one after onset of egg production. The distribution of deaths among the treatment groups was one (64 mg/kg diet) and one (160 mg/kg diet). There were no treatment-related effects on body weight, body weight change, feed consumption, survival and behavioural endpoints. No substance related abnormalities were noted at necropsy. There were no treatment-related effects on any reproductive parameter. Mean body weights of hatchlings and 14-day old chicks were not affected at any dose level.

Table 1. Results of a reproductive study on bobwhite quail (21-week exposure period)

Nominal concentration (mg/kg diet)	0	64	160	400
Mean measured concentration (mg a.s./kg diet)	-(A)	61	157	403
Mean parental food consumption (g/bird/day)	21	22	20	20
Mean body weight change males (%)	+2.4	+2.9	+3.9	+4.0
Mean body weight change females (%)	+12	+14	+14	+17
Mean egg production (eggs/cage)	36	36	32	40
Total number of eggs set	497	464	403	565
Mean eggshell thickness (mm)	0.21	0.21	0.21	0.21
Percent of eggs laid that were cracked (%)	3	3	2	1
Percent of eggs set that were fertile (%)	82	90	91	93
21-day embryos as percentage of fertile eggs (%)	98	98	98	99
21-day embryos as percentage of eggs set (%)	79	89	92	94
Number hatchlings as percent of 21-day embryos (%)	94	95	95	94
Number hatchlings as percent of fertile eggs (%)	92	94	93	92
Number hatchlings as percent of eggs set (%)	77	83	84	87
Number 14-day chicks as percent of hatchlings (%)	89	94	82	83
Number 14-day chicks as percent of fertile eggs (%)	83	87	80	81
Number 14-day chicks as percent of eggs set (%)	69	78	69	72
Number 14-day chicks per hen	21	24	20	27
Mean body weight (g) of hatchlings	5.8	5.8	5.8	6.0
Mean body weight (g) of 14-day chicks	23	22	23	23

(A) Not measured

Validity criteria fulfilled:

yes

Conclusions:

In a reproductive toxicity study in birds, performed in accordance with EPA 71-4, the NOEC in bobwhite quail exposed to the test substance in the diet was determined to be 400 mg/kg diet (mean measured concentration 403 mg/kg diet, equivalent to 39.2 and 35.9 mg/kg bw/day for males and females, respectively)

Executive summary:

In this reproductive toxicity study performed according to EPA 71 -4 guidance in compliance with GLP, 29-weeks-old bobwhite quails were exposed to the test substance in the diet for 21 weeks. Nominal diet concentrations were 0 (control), 64, 160 and 400 mg/kg diet. In week 20, nominal diet concentrations were inadvertently changed to 0, 177, 443 and 1108 mg/kg diet. There were 16 pairs per dose level, one pair per cage. The birds were maintained in a controlled environment test room. Eggs were collected daily from the onset of egg production (week 13). Hatchlings were leg banded and maintained for 14 days in cages according to pen.

Reproduction endpoints were determined with eggs produced during the last 9 weeks of the study. Reproduction endpoints were: number of eggs laid, number of eggs cracked, eggshell thickness, number of fertile eggs, number of live 21-day-old embryos, number of hatchlings, hatchling weight, number of 14-day-old survivors, and chick weights at day 14 of age. For eggshell thickness determination, eggs were collected weekly throughout the egg laying period.

No treatment-related effects were observed in the adult birds. One bird died before and one after onset of egg production. The distribution of deaths among the treatment groups was one (64 mg/kg diet) and one (160 mg/kg diet). There were no treatment-related effects on body weight, body weight change, feed consumption, survival and behavioural endpoints. No substance related abnormalities were noted at necropsy. There were no treatment-related effects on any reproductive parameter. Mean body weights of hatchlings and 14-day old chicks were not affected at any dose level.

The NOEC in bobwhite quail exposed to the test substance in the diet was determined to be 400 mg/kg diet (mean measured concentration 403 mg/kg diet, equivalent to 39.2 and 35.9 mg/kg bw/day for males and females, respectively).

Description of key information

All available data was assessed and the study representing the worst-case effects is included here as key studies. The key study is selected for the CSA. The other studies are included as supporting information.

19-wk NOEL = 17.7 and 18.4 mg/kg bw/day (160 mg/kg diet) for males and females, respectively, Anas platyrhynchos, reproduction parameters, EPA 71 -4, Beavers 1990a

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for birds:

160 mg/kg food

Additional information

EPA 71 -4, Beavers 1990a

In a reproductive toxicity study performed according to EPA 71-4 guidance and in compliance with GLP, 31-weeks-old mallard ducks were exposed to the test substance in the diet for 19 weeks. Nominal diet concentrations were 0 (control), 64, 160 and 400 mg/kg diet. There were 16 pairs per dose level, one pair per cage. The birds were maintained in a controlled environment test room. Eggs were collected daily from the onset of egg production (week 11). Hatchlings were leg banded and maintained for 14 days in cages according to pen. Reproduction endpoints were determined with eggs produced during the last 9 weeks of the study. Reproduction endpoints were: number of eggs laid, number of eggs cracked, eggshell thickness, number of fertile eggs, number of live 21-day-old embryos, number of hatchlings, hatchling weight, number of 14-day-old survivors, and chick weights at day 14 of age. For eggshell thickness determination, eggs were collected weekly throughout the egg laying period.

No treatment-related effects were observed in the adult birds. One bird died before and two birds died after onset of egg production during the exposure period of the study. The distribution of deaths among the treatment groups was one (control), one (160 mg/kg diet) and one (400 mg/kg diet). There were no treatment-related effects on body weight, body weight change, feed consumption, survival and behavioural endpoints. No substance related abnormalities were noted at necropsy.

The number of hatchlings as percentage of eggs set, fertile eggs and 21-day embryos was reduced at 400 mg/kg diet (not statistically significant), resulting in a reduction of the number of 14-day old chicks per hen and as percentage of eggs set and fertile eggs (not statistically significant). The number of 14-day old chicks as percentage of hatchlings was significantly increased at 64 and 160 mg/kg diet. This was a function of 10 incidental mortalities in the control group due to a feeder malfunction, and therefore not considered to be related to treatment. Mean body weights of hatchlings and 14-day old chicks were not affected at any dose level. 

Based on reduced hatchability at 400 mg/kg diet, the NOEC in mallard duck exposed to the test substance in the diet was 160 mg/kg diet (mean measured concentration 146 mg/kg diet after correction for purity of the test substance, equivalent to 17.7 and 18.4 mg/kg bw/day for males and females, respectively).

Supporting studies

EPA 71 -1, Roberts 1982a

In this study on the acute oral toxicity of the test substance to young adult bobwhite quail (Colinus virginianus) performed following the EPA 71-1 guidance and in compliance with GLP, five male and five females per dose level (fasted overnight) were orally dosed with the test substance at dose levels of 0, 2867, 3584, 4480, 5600 and 7000 mg/kg bw, followed by a fourteen day observation period. No birds died in the control group. One bird died at 3584 mg a.s./kg. Birds were observed to be quiet at all dose levels during the first week. Body weight gain was slightly reduced at all dose levels during days 1-3. Food consumption was not affected and no abnormalities were noted at necropsy.

The acute oral LD50 for bobwhite quail is >7000 mg/kg bw.

EPA 71 -1, Roberts 1982b

In this study on the acute oral toxicity of the test substance to young adult Mallard ducks (Anas platyrhynchos) performed following the EPA 71-1 guidance and in compliance with GLP, five male and five females per dose level (fasted overnight) were orally dosed with the test substance at dose levels of 0, 1229, 1536, 1920, 2400 and 3000 mg/kg bw, followed by a fourteen day observation period. No birds died in the control group and any treatment group. No treatment related effects on behaviour were observed at any dose level. Body weight gain and food consumption were not affected and no abnormalities were noted at necropsy. The acute oral LD50 for mallard duck is >3000 mg/kg bw.

EPA 71 -2, Campbell 1993

In this study performed following the EPA 71-2 guidance and in compliance with GLP, five groups of 10-day old Bobwhite quails of indeterminate sex (10/group) were fed the test substance in the diet at analytically confirmed concentrations of 562, 1000, 1780, 3160 and 5620 mg/kg diet, respectively, for a period of 5 days followed by a 3-day recovery period. Four control groups of ten 10-day old Bobwhite quails received feed treated with the vehicle (corn oil).

Following test initiation and continuing until termination, all birds were observed at least twice daily. A record was maintained of all mortality, signs toxicity and abnormal behaviour. Body weights by group were measured at the initiation of the test, on Day 5, and at the termination of the test on Day 8. Average estimated feed consumption was determined in each test and control group for the exposure period Days 0-5, and for the post-exposure observation period, Days 6-8. Feed consumption was determined by the change in the weight of the feed presented to the birds over a given period of time.

One bird died in one of the control groups (Day 8). No further birds died during the study. No clinical signs were noted. Body weight gain was reduced at 3160 and 5620 mg/kg diet. Feed consumption was not affected at any dose level.

The 5-day dietary LC50 was >5620 mg/kg diet, equivalent to >2576 mg/kg bw/day. The 5-day NOEC was 1780 mg/kg diet, equivalent to 872 mg/kg bw/day.

EPA 71 -2, Roberts 1982c

In this study on the dietary toxicity of the test substance, performed following the EPA 71-2 guidance and in compliance with GLP, four groups of 15-day old Bobwhite quails of indeterminate sex (10/group) were fed the test substance in the diet at analytically confirmed concentrations of 100, 1000, 5000 and 20000 mg/kg diet, respectively, for a period of 5 days followed by a 6-day recovery period. Three control groups of ten 15-day old Bobwhite quails received untreated feed. No birds died in the control groups. Four and nine birds died at 100 and 20000 mg/kg diet, respectively. No further birds died during the study. The mortality at 100 mg/kg diet was considered not to be treatment-related, since no mortality was observed at the next two higher dose levels. No treatment related clinical signs were noted. Body weight gain and feed consumption were not affected at any dose level.The 5-day dietary LC50 is approximated as 10000 mg/kg diet, equivalent to 1356 mg/kg bw/day, corresponding to the geo-mean of diet concentrations with 'no mortalities' (5000 mg/kg diet) and '90 % mortality (20000 mg/kg diet). The 5-day NOEC was 5000 mg/kg diet, equivalent to 720 mg/kg bw/day.

EPA 71 -2, Roberts 1982d

In this study on the dietary toxicity of the test substance, performed following the EPA 71-2 guidance and in compliance with GLP, four groups of 13-day old Mallard ducks of indeterminate sex (10/group) were fed the test substance in the diet at analytically confirmed concentrations of 100, 1000, 5000 and 20000 mg/kg diet, respectively, for a period of 5 days followed by a 6-day recovery period. Three control groups of each ten Mallard ducks received untreated feed. No birds died in the control groups and in any treatment groups. No treatment related clinical signs were noted. Body weight gain was reduced at 20000 mg/kg diet. Feed consumption was not affected at any dose level. The 5-day dietary LC50 was >20000 mg/kg diet, equivalent to >5885 mg/kg bw/day. The 5-day NOEC was 5000 mg/kg diet, equivalent to 1251 mg/kg bw/day.

EPA 71 -4, Beavers 1990b

In this reproductive toxicity study performed according to EPA 71 -4 guidance in compliance with GLP, 29-weeks-old bobwhite quails were exposed to the test substance in the diet for 21 weeks. Nominal diet concentrations were 0 (control), 64, 160 and 400 mg/kg diet. In week 20, nominal diet concentrations were inadvertently changed to 0, 177, 443 and 1108 mg/kg diet. There were 16 pairs per dose level, one pair per cage. The birds were maintained in a controlled environment test room. Eggs were collected daily from the onset of egg production (week 13). Hatchlings were leg banded and maintained for 14 days in cages according to pen.

Reproduction endpoints were determined with eggs produced during the last 9 weeks of the study. Reproduction endpoints were: number of eggs laid, number of eggs cracked, eggshell thickness, number of fertile eggs, number of live 21-day-old embryos, number of hatchlings, hatchling weight, number of 14-day-old survivors, and chick weights at day 14 of age. For eggshell thickness determination, eggs were collected weekly throughout the egg laying period.

No treatment-related effects were observed in the adult birds. One bird died before and one after onset of egg production. The distribution of deaths among the treatment groups was one (64 mg/kg diet) and one (160 mg/kg diet). There were no apparent treatment-related effects upon reproductive parameters at any of the concetrations tested and also no effects on body weight, body weight change, feed consumption, survival and behavioural endpoints. No substance related abnormalities were noted at necropsy.

The NOEC in bobwhite quail exposed to the test substance in the diet was 400 mg/kg diet (mean measured concentration 403 mg/kg diet, equivalent to 39.2 and 35.9 mg/kg bw/day for males and females, respectively).