Ammonium 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)propanoate

Toxicity to birds

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 206 (Avian Reproduction Test)

Deviations:

no

Remarks:

The study was conducted according to the test guidelines in effect at the time of study conduct.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.2300 (Avian Reproduction Test)

Deviations:

no

Remarks:

The study was conducted according to the test guidelines in effect at the time of study conduct.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 002

Cas Number:

62037-80-3

Details on test material:

- Purity: 84%

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

no

Details on preparation and analysis of diet:

FOOD AND WATER
- The basal diet fed to both adults and offspring was formulated to Wildlife International, Ltd. specifications by Cargill Animal Nutrition, Shippensburg, PA
- Water was supplied by the town of Easton, MD public water supply.

HOMOGENEITY AND STABILITY OF TEST MATERIAL IN DIET
- Test diets were prepared by mixing the test substance into a premix that was used for weekly preparation of the final diet. Control diet and each of the three treated diets were prepared weekly and presented to the birds on Wednesday of each week. Dietary concentrations were not adjusted for purity of the test substance and are presented as parts per million (ppm).

- Homogeneity of the test substance in the diet was evaluated by collecting six samples from each of the treated diets and one sample from the control diet on Day 0 of Week 1. Control and treatment group diet samples were also collected from the feeders on Day 7 of Week 1 to assess stability of the test substance under actual test conditions. Additionally, samples were collected from the control and treatment group diets during Weeks 2, 3, 4, 8, 12, 16 and 20 of the test to measure/verify test concentrations.

- How often was stability tested: Analysis of diet samples collected from feeders after being held at ambient temperature for 7 days averaged 109%, 114% and 104% of the Day 0 values for the 100, 500, and 1000 ppm test concentrations, respectively
- Nominal concentration: 100, 500 and 1000 ppm
- Concentration analyzed: (mean measured values from weeks 2 - 20) 90.8 ± 9.20, 476 ±72.6 and 950 ± 127 ppm
- % of nominal: 91%, 95% and 95%
- Mixing procedure adequate and variance between nominal and actual dosage acceptable (yes/no): yes

Test organisms

Test organisms (species):

Colinus virginianus

Details on test organisms:

TEST ORGANISM
- Common name: northern bobwhite quail
- Source: M & M Quail Farm, Gillsville, GA
- Age at test initiation: approximately 31 weeks
- Weight at test initiation: ranged in weight from 172 to 242 grams
- Sexes used: male/female

Study design

Total exposure duration (if not single dose):

20 wk

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

16

Control animals:

yes, concurrent no treatment

Nominal and measured doses / concentrations:

- Nominal concentration: 0, 100, 500 and 1000 ppm
- Concentration analyzed: (mean measured values from weeks 2 - 20) 90.8 ± 9.20, 476 ±72.6 and 950 ± 127 ppm

Details on test conditions:

ACCLIMATION
- Acclimation period: 10 weeks
- Health (any disease or mortality observed): none
- Fasting period before study: no

PEN SIZE AND CONSTRUCTION MATERIALS
- Description: 25 X 51 cm
- Materials: Pen constructed of wire mesh and galvanized sheeting, sisal ropes were added to each pen for animal enrichment from the time of photostimulation to termination.
- Compliant to good husbandry practices: yes
- Suitable to avoid crowding stress: yes
- Caging: one pair per pen

NO. OF BIRDS PER REPLICATE
- For control: 2
- For treated: 2

NO. OF REPLICATES PER GROUP
- For control: 16
- For treated: 16

TEST CONDITIONS
- Temperature: Adults - 21.1°C (+/- 0.9°C SD); Chicks - 26.6 ± 2.1°C
- Relative humidity (%): Mean = 60% (+/- 14% SD)
- Photoperiod: 8 hour light per day during acclimation and first seven weeks of test; 17 hours of light per day during remainder of test (approximately 343 lux)
- Ventilation: 15 room air volumes every hour
- Brooder temperature: Thermostats in the brooding compartment of each pen were set to maintain a temperature of approximately 38° C from the time of hatching until the birds were 14 days of age.

RANGE FINDING STUDY
Northern bobwhite quail were exposed to the test substance at dietary concentrations of 0, 10, 100 and 1000 ppm over a six-week period. There were no treatment-related mortalities, overt signs of toxicity or treatment-related effects upon body weight or feed consumption at any of the test concentrations. Additionally, there were no treatment-related effects upon any of the reproductive parameters measured at the 10 or 100 ppm test concentrations. At the 1000 ppm concentration there were slight reductions in viability of embryos, and reductions in numbers of hatchlings and 14-day old survivors as percentages of eggs set and maximum set that were likely treatment related. The NOEC for northern bobwhite quail exposed to the test substance in the diet during the study was 100 ppm.

Examinations

Details on examinations and observations:

MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: During the study, all adult birds were observed daily for signs of toxicity or abnormal behavior.
- Remarks: Additionally, all offspring were observed daily from hatching until 14 days of age. A record was maintained of all mortalities and clinical observations.

BODY WEIGHT
- Time schedule for examinations: at test initiation, at the end of Weeks 2, 4, 6, 8 and at adult termination
- Remarks: Body weights were not measured during egg laying because of the possible adverse effects handling may have on egg production.

FOOD CONSUMPTION (if feeding study)
- Time schedule for examinations: Feed consumption for each pen was measured weekly throughout the test.

PATHOLOGY
- Dose groups that were examined: All surviving adults were subjected to gross necropsy following adult termination.

Details on reproductive parameters:

REPRODUCTION
- Parameters: Reproductive parameters were measured beginning at the onset of egg laying.
Total eggs laid/group, weekly
Eggs cracked, weekly
Eggs set
Viable embryos, eggs were candled on day 11 (northern bobwhite quail) of incubation to determine embryo viability and on day 21 to determine embryo survival
Live three-week embryos
Hatchlings
14-day-old survivors
Eggs laid/hen
Eggs laid/hen/day
14-day-old survivors/hen
Egg shell thickness (mm), weekly
Hatchling body weight (g)
14-day old chick body weight (g)

Results and discussion

Effect levels

Effect levels 1

Duration (if not single dose):

20 wk

Dose descriptor:

NOEC

Effect level:

1 000 other: ppm

Conc. / dose based on:

test mat.

Basis for effect:

other: mortality and reproduction

Remarks on result:

other: equivalent to 84.5 mg/kg/day

Effect levels 2

Duration (if not single dose):

20 wk

Dose descriptor:

LOEC

Effect level:

> 1 000 other: ppm

Conc. / dose based on:

test mat.

Basis for effect:

other: mortality and reproduction

Remarks on result:

other: equivalent to 84.5 mg/kg/day

Mortality and sub-lethal effects:

MORTALITY and CLINICAL SIGNS
No mortalities occurred during the study.
No overt signs of toxicity were observed at any of the test concentrations. Incidental clinical observations noted during the test included those that normally are associated with injuries and penwear. Such signs included feather loss, foot, head, neck and back lesions, bumps, bruising and swelling. Additional clinical observations included lameness, a ruffled appearance, wing droop, ventral head curl and the female in Pen 445 of the 500 ppm treatment group was noted as thin during Week 20 of the test. Except for incidental findings, all birds appeared normal throughout the study.

PATHOLOGY
All surviving adults were subjected to gross necropsy. All necropsy findings observed were considered unrelated to treatment.

BODY WEIGHT
There were no apparent treatment related effects upon adult body weight at any of the test concentrations. No statistically significant differences between the control group and the 100, 500, or 1000 ppm treatment groups were observed at any of the body weight intervals.

FOOD CONSUMPTION
There were no apparent treatment related effects upon feed consumption at any of the test concentrations. No statistically significant differences between the control group and the 100, 500 or 1000 ppm treatment groups were observed at any of the feed consumption intervals.

Effects on reproduction:

REPRODUCTIVE RESULTS
There were no treatment-related effects upon reproductive performance at any of the test concentrations. When compared to the control group, there were no statistically significant differences in any of the reproductive parameters measured in the 100, 500 or 1000 ppm treatment groups.

EGG SHELL THICKNESS
There were no apparent treatment related effects upon shell thickness at any of the concentrations tested. When compared to the control group, there were no statistically significant differences in shell thickness in the 100, 500 or 1000 ppm treatment groups.

OFFSPRING BODY WEIGHTS
There were no apparent treatment related effects upon offspring body weight at any tested concentrations. When compared to the control group, there were no statistically significant differences in the body weight of hatchlings or 14-day old survivors from the 100, 500 or 1000 ppm treatment groups.

Further details on results:

BLOOD ANALYSIS
None of the control samples showed any indication of the presence of the test substance or of the presence of a co-eluting substance at the characteristic retention time of the test substance. Blood samples were collected from birds that were exposed to diets fortified with the test substance at 100, 500 and 1000 ppm nominal test concentrations. The means and standard deviations for residues found in blood at the three test concentrations were 1220 ± 714 ppb, 2678 ± 2336 ppb and 5110 ± 5346 ppb, respectively. Blood samples collected from the offspring of the adult pair had no measurable values above or at the LOQ

LIVER ANALYSIS
None of the control samples showed any indication of the presence of the test substance or of the presence of a co-eluting substance at the characteristic retention time of the test substance. Liver samples were collected from birds of corresponding blood samples that were exposed to diets fortified with the test substance at 100, 500 and 1000 ppm test concentrations. The means and standard deviations for residues found in liver at the three test concentrations were 438 ± 301 ppb, 991.1 ± 858.6 ppb and 2008 ± 1730 ppb, respectively. Liver samples collected from the offspring of the adult pairs had no measurable values above or at the LOQ in adults exposed at the 100 ppm diet. The means and or standard deviations for residues found in liver at the two test concentrations of 500 and 1000 were 10.2 ± 4.03 ppb and 6.57 ± 1.09 ppb, respectively for samples that had residues above the LOQ.

EGG ALBUMIN ANALYSIS
None of the control samples showed any indication of the presence of the test substance or of the presence of a co-eluting substance at the characteristic retention time of the test substance. Albumin samples were collected from eggs in which birds were exposed to diets fortified with the test substance at 100, 500 and 1000 ppm test concentrations. The means and standard deviations for residues found in albumin at the three test concentrations were 56.2 ± 24.5 ppb, 240 ± 87.4 ppb and 412 ± 189 ppb, respectively

EGG YOLK ANALYSIS
None of the control samples showed any indication of the presence of the test substance or of the presence of a co-eluting substance at the characteristic retention time of the test substance. Yolk samples were collected from eggs in which birds were exposed to diets fortified with the test substance at 100, 500 and 1000 ppm test concentrations. The means and standard deviations for residues found in yolks at the three test concentrations were 1361 ± 524 ppb, 7118 ± 1128 ppb and 12448 ± 3120 ppb, respectively.

EGGSHELL ANALYSIS
The control samples showed some indication of the presence of the test substance or of the presence of a co-eluting substance at the characteristic retention time of the test substance These values were above the reported LOQ Eggshell samples were collected from eggs in which birds were exposed to diets fortified with the test substance at 100, 500 and 1000 ppm test concentrations. The means and standard deviations for residues found in eggshells at the three test concentrations were 105 ± 170 ppb, 83.4 ± 17.1 ppb and 216 ± 95.5 ppb, respectively.

EGG MEMBRANE ANALYSIS
The control sample was a composite of membranes from egg lots where most of the egg albumin, yolk and shells analysis was done. The control sample showed a slight indication of the presence of the test substance or of the presence of a coeluting substance at the characteristic retention time of the test substance. This value was below the reported LOQ. Egg membrane samples were a composite collected from eggs lots in which most of the other egg albumin, yolk and shells analyses occurred and that birds were exposed to diets fortified with the test substance at 100, 500 and 1000 ppm test concentrations. The measured values for those samples at the three test concentrations were 37.1 ppb, 170 ppb and 128 ppb, respectively.

Reported statistics and error estimates:

Upon completion of the test, an analysis of variance (ANOVA) was performed to determine statistically significant differences between groups. Dunnett's multiple comparison procedure was used to compare the three treatment means with the control group mean and assess the statistical significance of the observed differences. Percentage data were examined using Dunnett's method following arcsine square root transformation.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

This study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
NOEC = 1000 ppm
LOEC > 1000 ppm

Executive summary:

The objective of this study was to evaluate the effects upon the adult northern bobwhite quail (Colinus virginianus) of dietary exposure to the test substance over a five-month period. Effects were evaluated on adult health, weight gain and feed consumption. In addition, the effects of adult exposure to the test substance were evaluated on the number of eggs laid, normal development of eggs, viability of the embryos, percent hatchability, offspring survival and egg shell thickness.

There were no treatment-related mortalities, overt signs of toxicity or treatment related effects upon body weight or feed consumption at any tested concentrations. Additionally, there were no treatment-related effects upon reproductive performance parameters measured at the 100, 500 or 1000 ppm test concentrations. The no-observed-effect concentration (NOEC) for northern bobwhite quail exposed to the test substance in the diet during this study was 1000 ppm (equivalent to 84.5 mg/kg/day), the highest nominal test concentration and the lowest observed effect concentration (LOEC) was > 1000 ppm (equivalent to > 84.5 mg/kg/day).