pymetrozine (ISO); (E)-4,5-dihydro-6-methyl-4-(3-pyridylmethyleneamino)-1,2,4-triazin-3(2H)-one

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

Study period:

17 Oct 1990 to 31 Oct 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: EPA – FIFRA Pesticides Assessment Guidelines, Subdivision E, Hazard Evaluation; Wildlife and Aquatic Organisms, Series 71 – Avian and mammalian testing. Avian Single-dose oral LD50 test. Paragraph 71-1

Version / remarks:

1982

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

14 d

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Mortality and growth are summarised in the table below.

 

Summary of effects of the test substance on mortality and growth of Mallard duck (Anas platyrhynchos) following acute oral exposure.

Group	Cumulative mortality		sex	Mean weight gain (g/bird)				Food consumption (g/animal/day)
				Days 0 to 3	Days 4 to 7	Days 8 to 14	Days 0 to 14	Days -7 to -1	Days 0 to 3	Days 4 to 7	Days 8 to 14
1 0 mg/kg (control)	0		male	111	6	-3	114	79	77	64	86
	0		female	80	15	15	110	79	77	64	86
2  2000 mg/kg	0		male	83	35	1	119	79	68	75	86
	0		female	68	37	5	110	79	68	75	86
3  2000 mg/kg	0		male	111	6	-3	114	79	77	64	86
	0		female	80	15	15	110	79	77	64	86

 

There were no mortalities in the control group and all birds in the control group were normal in appearance and behaviour for the duration of the test. There were no mortalities or treatment related clinical signs in the treatment groups. No significant pathological changes were observed in any birds.The 14-day acute oral LD50 for mallard duck exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the only concentration tested.

Validity criteria fulfilled:

yes

Conclusions:

The 14-day acute oral LD50 for mallard duck exposed to test substance was determined to be greater than 2000 mg test substance/kg body weight, the only concentration tested.

Executive summary:

The acute oral toxicity of the test item to the Mallard duck (Anas platyrhynchos) was determined using the GLP compliant study according to US EPA FIFRA 71-1 guideline. The test organism were treated, in duplicates, with 2000 mg test substance/kg body weight and control (vehicle only) for 14 days.

As the test was designed as a limit test it consisted of one test concentration. A control group was also used. Mallard ducks, approximately 24 weeks old, and in good health, were randomly assigned to test groups and fasted for a minimum of 16 hours prior to test initiation. The test substance was administered by oral intubation to ten birds of each sex at 2000 mg test substance/kg body weight as a suspension in 2 % v/w carboxymethylcellulose in OM-water, as a single oral dose. A control group, five birds per sex, was dosed with the vehicle at 20 mL/kg. The dosing was followed by a fasting period of 16 hours. The birds were housed outdoors by treatment group (5 females, 5 males per pen) in wire net cages without bottoms measuring 4 x 4 x 2 m. During the test a grain mixture of wheat plus water was provided ad libitum. The birds were observed continuously until symptoms existed, then daily, mortalities were noted during inspection of symptoms. Individual body weights were measured on day -7 (start of acclimation), at test initiation and on days 3, 7 and 14. Food consumption was measured for the periods Day -7 to -1, 0-3, 4-7, and 8-14. At test termination, post-mortem examination was carried out.

There were no mortalities in the control group and all birds in the control group were normal in appearance and behaviour for the duration of the test. There were no mortalities or treatment related clinical signs in the treatment groups. No significant pathological changes were observed in any birds. The 14-day acute oral LD50 for mallard duck exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the only concentration tested.

Reference 2

Endpoint:

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

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

26 Feb 1991 to 12 Mar 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: U.S. Environmental Protection Agency. 1982. Avian Single-Dose Oral LD50 Test Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation; Wildlife and Aquatic Organisms, subsection 71-1

Version / remarks:

incorporating draft guidelines dated 1988

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Coturnix coturnix japonica

Duration (if not single dose):

14 d

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

14 d

Dose descriptor:

NOEL

Effect level:

2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Mortality, growth and food consumption are summarised in the table below.

 

Summary of effects of the test substance on mortality and growth of Japanese quail (Coturnix coturnix japonica) following acute oral exposure.

Group	Treatment (mg test substance/ kg)	Cumulative  mortality	Sex	Mean weight gain (g/bird)				Food consumption (g/animal/day)
				Days 0 to 3	Days 4 to 7	Days 8 to 14	Days 0 to 14	Days -7 to -1	Days 0 to 3	Days 4 to 7	Days 8 to 14
1	0 (control)	0	male	5	4	5	15	38	33	35	34
	0 (control)	0	female	4	8	6	18	38	33	35	34
2	2000	0	male	6	6	7	19	38	32	34	35
	2000	0	female	3	5	6	14	38	32	34	35
3	2000	0	male	5	7	5	17	37	34	36	35
	2000	0	female	3	14	5	22	37	34	36	35

 

There were no mortalities in the control group or the treatment groups, and all birds were normal in appearance and behaviour for the duration of the test. There were no signs of toxicity observed in any group, and no dose-related differences in body weight or food consumption were observed. At test termination, no abnormalities were observed during gross macroscopic post mortem examination. The oral LD50 for Japanese quail exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the only concentration tested.

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test, the oral LD50 for Japanese quail exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the only concentration tested.

Executive summary:

The acute oral toxicity of the test item to the Japanese quail (Coturnix coturnix japonica) was determined using the GLP compliant study according to US EPA FIFRA 71-1 guideline. Japanese quail (Coturnix coturnix japonica) were treated, in duplicates, with 2000 mg test substance/kg body weight and control (vehicle only) for 14 days. As the test was designed as a limit test it consisted of one test concentration. A control group was also used. Japanese quail, 6 weeks old and in good health, were randomly assigned to test groups and fasted for a minimum of 16 hours prior to test initiation. The test item was administered by oral intubation to ten birds of each sex per group at 2000 mg test substance/kg body weight as a suspension in 1 % v/w carboxymethyl cellulose, as a single dose. A control group, five birds per sex, was dosed with the vehicle at 20 mL/kg. The dosing was followed by a fasting period of 16 hours. The birds were housed indoors by treatment group (5 females, 5 males per pen) in cages measuring 95 x 45 x 50 cm, made of wire mesh, and containing an automatic drinker and plate feeder. During testing all test birds were fed ad libitum with a factory mixed standard diet, and had free access to water. The birds were observed daily for toxicological responses throughout the test. A record was maintained of mortality, signs of toxicity, and health. Individual body weights were measured on Days -7, 0, 3, 7 and 14. Group feed consumption was determined over Days -7 to -1, 0 to 3, 4 to 7 and 8 to 14. At test termination, all birds were examined for gross pathological changes.

There were no mortalities in the control group or the treatment groups, and all birds were normal in appearance and behaviour for the duration of the test. There were no signs of toxicity observed in any group, and no dose-related differences in body weight or food consumption were observed. At test termination, no abnormalities were observed during gross macroscopic post mortem examination. The oral LD50 for Japanese quail exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the only concentration tested.

Reference 3

Endpoint:

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

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

24 Jun 1992 to 8 Jul 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: U.S. Environmental Protection Agency. 1982. Avian Single-Dose Oral LD50 Test Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation; Wildlife and Aquatic Organisms, subsection 71-1

Version / remarks:

incorporating draft guidelines dated 1988

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

14 d

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

14 d

Dose descriptor:

NOEL

Effect level:

2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Mortality, growth and food consumption are summarised in the tables below.

 

Summary of effects of the test substance on mortality and growth of male Bobwhite Quail (Colinus virginianus) following acute oral exposure.

	Mean weight gain (g)					Group mean consumption (g/animal /day)
Treatment (mg test substance/kg)	Cumulative mortality (n = 5)	Days -15 to -7	Days -7 to 0	Days 0 to 7	Days 7 to 14	Dayes -15 to -8	Days -7 to -1	Days 1 to 7	Days 8 to 14
Control	0	-3	-2	1	4	13	14	13	16
500	0	-2	-4	0	4	14	14	13	15
1000	0	-5	-3	0	3	14	13	13	15
2000	0	-5	-3	-2	3	16	15	12	20

 

 

Summary of effects of the test substance on mortality and growth of female Bobwhite Quail (Colinus virginianus) following acute oral exposure.

	Mean weight gain (g)					Group mean consumption (g/animal /day)
Treatment (mg test substance/kg)	Cumulative mortality (n = 5)	Days -15 to -7	Days -7 to 0	Days 0 to 7	Days 7 to 14	Dayes -15 to -8	Days -7 to -1	Days 1 to 7	Days 8 to 14
Control	0	-2	-6	6	3	14	13	14	15
500	0	-4	-1	3	2	13	13	13	14
1000	0	-4	-2	2	4	13	14	13	15
2000	0	-3	-2	-2	4	16	17	14	19

 

There were no mortalities in the control group and all birds in the control group were normal in appearance and behaviour for the duration of the test. There were no mortalities in any of the test item treatment groups. No regurgitation was noted after dosing, either in the control group or in any of the treatment groups. Signs of toxicity (subdued behaviour and unsteadiness) were noted in birds at all test item concentration levels approximately 2 hours after dosing. All birds had recovered by the beginning of Day 3. In the highest treatment group of 2000 mg test substance/kg, a slight body weight loss was observed in the period after dosing (Days 0 to 7) when compared to the control values. All other body weight changes were similar and no other treatment-related effects were observed. Food consumption was observed to be reduced in the highest treatment group (2000 mg test substance/kg) in the period Day 1 to 7. A compensatory increase was observed in the same group over Days 8 to 14. Food consumption data in the other treatment groups were similar to the control values. No abnormalities were detected in any bird examined post mortem at the termination of the study. The 14-day oral LD50 for northern bobwhite exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the highest concentration tested. The no-mortality concentration and the no-observed effect concentration were 2000 mg test substance/kg body weight.

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test, the 14-day oral LD50 for bobwhite quail exposed to the test item was determined to be greater than 2000 mg/kg body weight, the highest concentration tested. The no-mortality concentration and the no-observed effect concentration were 2000 mg test substance/kg body weight.

Executive summary:

The acute oral toxicity of the test item to the bobwhite quail was determined using the GLP compliant study according to US EPA FIFRA 71-1 guideline. Nominal concentrations Nominal: 500, 1000 and 2000 mg test item/kg body weight, alongside a control (corn oil) were used on bobwhite quail (Colinus virginianus) for 14 days. Bobwhite quail, greater than 16 weeks old and in good health, were randomly assigned to test groups and fasted for 21 hours prior to test initiation. The test substance was administered by oral intubation to five birds of each sex per group at 500, 1000 or 2000 mg test substance/kg body weight, as a single dose. A similar sized control group was dosed with corn oil. The birds were housed by dosage group and sex in tiered cages in a ventilated building. Each cage measured approximately 0.39 x 0.76 x 0.24 m, was made of stainless steel wire mesh, and contained an automatic drinker and two food hoppers. During the testing all test birds were fed ad libitum with standard test facility layer diet in pellet form, and had free access to water. The birds were observed at frequent intervals during the post treatment period and then daily for toxicological responses. A record was maintained of mortality, signs of toxicity, andhealth. Individual body weights were measured on Days -15, -7, 0 (immediately prior to dosing), 7 and 14. Group mean food consumption was determined over Days -15 to -8, -7 to -1, 1 to 7, and 8 to 14. Any bird that died during the study was examined post-mortem. At test termination, post-mortem examination was carried out on ten birds from the highest dose group and on ten control birds. The LD50 values and the 95 % confidence intervals could not be determined since no mortalities occurred during the study. No statistical analyses were reported to separate mean responses among treatment groups for the endpoints of food consumption and body weight.

There were no mortalities in the control group and all birds in the control group were normal in appearance and behaviour for the duration of the test. There were no mortalities in any of the test item treatment groups. No regurgitation was noted after dosing, either in the control group or in any of the treatment groups. Signs of toxicity (subdued behaviour and unsteadiness) were noted in birds at all test item concentration levels approximately 2 hours after dosing. All birds had recovered by the beginning of Day 3. In the highest treatment group of 2000 mg test substance/kg, a slight body weight loss was observed in the period after dosing (Days 0 to 7) when compared to the control values. All other body weight changes were similar and no other treatment-related effects were observed. Food consumption was observed to be reduced in the highest treatment group (2000 mg test substance/kg) in the period Day 1 to 7. A compensatory increase was observed in the same group over Days 8 to 14. Food consumption data in the other treatment groups were similar to the control values. No abnormalities were detected in any bird examined post mortem at the termination of the study.The 14-day oral LD50 for northern bobwhite exposed to the test substance was determined to be greater than 2000 mg test substance/kg body weight, the highest concentration tested. The no-mortality concentration and the no-observed effect concentration were 2000 mg test substance/kg body weight.

Reference 4

Endpoint:

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

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

20 May 1992 to 03 Jun 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: U.S. Environmental Protection Agency. 1982. Avian Single-Dose Oral LD50 Test Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation; Wildlife and Aquatic Organisms, subsection 71-1

Version / remarks:

incorporating draft guidelines dated 1988

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

14 d

Dose descriptor:

other: 50% emetic dose (ED50)

Effect level:

82.5 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

behaviour

Remarks:

emetic nature of compound

Duration (if not single dose):

14 d

Dose descriptor:

NOEL

Effect level:

31.25 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

behaviour

Remarks:

emetic nature of compound

Mortality and growth are summarised in the table below.

 

Summary of effects of the test substance on mortality and growth male Mallard duck (Anus platyrhynchos) following acute oral exposure

Treatment (mg test substance/kg)	Mean weight gain (g)					Group mean consumption (g/animal /day)
	Cumulative mortality (n = 5)	Days -15 to -7	Days -7 to 0	Days 0 to 7	Days 7 to 14	Dayes -15 to -8	Days -7 to -1	Days 1 to 7	Days 8 to 14
Control	1*	-21	-96	28	-7	43	60	63	43
31.25	0	-39	-16	-11	11	88	66	94	49
125	0	13	-41	31	-32	38	57	117	20
500	0	-135	22	-2	5	20	51	89	103
2000	0	-79	3	-14	7	48	89	91	83

* Bird found dead on Day 11; evidence of feather pecking was seen when bird examinedpost-mortem

 

Summary of effects of the test substance on mortality and growth female Mallard duck (Anus platyrhynchos) following acute oral exposure

Treatment (mg test substance/kg)	Mean weight gain (g)					Group mean consumption (g/animal /day)
	Cumulative mortality (n = 5)	Days -15 to -7	Days -7 to 0	Days 0 to 7	Days 7 to 14	Dayes -15 to -8	Days -7 to -1	Days 1 to 7	Days 8 to 14
Control	1*	-93	17	80	-27	75	91	131	109
31.25	0	-80	28	61	-86	108	26	137	120
125	0	54	-44	-8	-108	95	160	89	57
500	1*	-49	78	15	-44	50	97	134	120
2000	1*	-85	2	82	12	23	63	97	129

* The three mortalities occurred during the pre-dose acclimatisation period and were replaced

 

There was one post-treatment mortality among control group males but, on post mortem examination, the death was deemed not have been treatment related. All other birds in the posttreatment control group were normal in appearance and behaviour for the duration of the test. Following dosing, vomiting was observed in birds from treatment groups of ≥125 mg test substance/kg body weight. No other signs of toxicity were observed. All birds appeared normal from the end of Day 1 onwards. Body weight changes and food consumption were variable in all groups and there was no evidence of any treatment-related effects. No abnormalities were detected in any bird examined at termination. It was not possible to calculate the LD50 value due to the emetic nature of the compound. The 50 % emetic dose (ED50) was found to be 82.5 mg test substance /kg body weight, and the no effect level was 31.25 mg test substance/kg body weight.

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test with the test substance and the Mallard duck, it was not possible to calculate the LD50 value due to the emetic nature of the compound. The 50 % emetic dose (ED50) was found to be 82.5 mg test substance/kg body weight, and the no effect level was 31.25 mg test substance/kg body weight.

Executive summary:

The acute oral toxicity of the test item to the Mallard duck (Anas platyrhynchos) was determined using the GLP compliant study according to US EPA FIFRA 71-1 guideline. The test organism were treated, in duplicates, with 31.25, 125, 500 and 2000 mg/kg body weight, alongside a control (corn oil)for 14 days.

Mallard ducks, approximately 11 months old, and in good health, were randomly assigned to test groups and fasted for 21 hours prior to test initiation. The test substance was administered by oral intubation to five birds of each sex per group at 31.25, 125, 500 or 2000 mg/kg body weight, as a single oral dose. A similar sized control group was dosed with corn oil. The birds were housed in galvanised steel pens with a solid concrete floor, five birds per cage. Each pen measured approximately 2.22 × 1.68 m and contained a food hopper and an automatic drinker, with straw provided as bedding material. The test substance formulations were prepared immediately prior to dosing and homogeneity in the vehicle was determined by HPLC analysis. During the test each bird was fed a standard laboratory layer diet in pellet form ad libitum, and had free access to tap-water. The birds were observed immediately after dosing and daily thereafter. Individual body weights were measured on Days -15, -7, 0 (immediately prior to dosing), 7 and 14. Group mean food consumption was calculated over Days -15 to -8, -7 to -1, 1 to 7 and 8 to 14. Any bird that died during the study was examined post-mortem. At test termination, post-mortem examination was carried out on ten birds from the highest dose group and on ten control birds. The LD50 values were not calculated due to the emetic nature of the compound, so a 50 % emetic dose was determined using the method of Thompson and Weil.

There was one post-treatment mortality among control group males but, on post mortem examination, the death was deemed not have been treatment related. All other birds in the posttreatment control group were normal in appearance and behaviour for the duration of the test. Following dosing, vomiting was observed in birds from treatment groups of ≥125 mg test substance/kg body weight. No other signs of toxicity were observed. All birds appeared normal from the end of Day 1 onwards. Body weight changes and food consumption were variable in all groups and there was no evidence of any treatment-related effects. No abnormalities were detected in any bird examined at termination. It was not possible to calculate the LD50 value due to the emetic nature of the compound. The 50 % emetic dose (ED50) was found to be 82.5 mg test substance /kg body weight, and the no effect level was 31.25 mg test substance/kg body weight.

Reference 5

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

12 Mar 1992 to 23 Mar 1992

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)

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

5 d

Dose descriptor:

LC50

Effect level:

> 5 200 other: ppm

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

5 d

Dose descriptor:

NOEL

Effect level:

2 600 other: ppm

Conc. / dose based on:

test mat.

Basis for effect:

body weight

Mortalities occurred at 0 and 2600 ppm (one bird at each level). All surviving birds remained in good health throughout the study although there was evidence of aggressive behaviour at 0 ppm. There were no clinical signs of toxicity. Food consumption and bodyweight increase were reduced during the treatment period at 5200 ppm. There were no treatment-related abnormalities observed post-mortem at termination. The mortality effect at 2600 was not regarded as substance-related. Apart from that the only effects were seen on body weight and feed consumption at 5200 ppm. Therefore, the LD50 was determined to be > 5200 ppm and the NOEL was determined to be 2600 ppm.

 

Analytical results

Nominal inclusion (ppm)	Mean analysed concentration (ppm)	Relative mean error (%)
163	164	0.6
325	318	-2.2
650	625	-3.8
1300	1300	0
2600	2550	-1.9
5200	5130	-1.3

Validity criteria fulfilled:

yes

Conclusions:

There were no treatment-related abnormalities observed post-mortem at termination. The LD50 value of the test substance to the Bobwhite quail was in excess of 5200 ppm. The no observed effect level in this study was 2600 ppm.

Executive summary:

A study was performed to determine the subacute dietary toxicity (LC50) of the test substance to the Bobwhite quail. The method followed was based on that given in the United States Environmental Protection Agency (EPA) Pesticide Assessment Guidelines, Subdivision E, Hazard Evaluation, Wildlife and Aquatic Organisms, Series 71 - Avian and Mammalian Testing, § 71-2 Avian dietary LC50 test, dated October 1982 and draft revised guideline, dated March 1988. Groups of 10 young Bobwhite quail chicks were offered diet containing 163, 325, 650, 1300, 2600 or 5200 ppm test item. Two similar sized control groups were offered basal diet alone. Test diets were introduced when the birds were 11 days old and were offered to the birds for 5 days. Test diets were then replaced with basal diet and the birds observed for a further 3 days. Observations, including mortality, clinical signs, bodyweights and food consumption were made during the study. The birds were then killed and 10 control birds together with 10 birds from the highest dose level groups in which there were survivors subjected to a gross post-mortem examination. Mortalities occurred at 0 and 2600 ppm (one bird at each level). All surviving birds remained in good health throughout the study although there was evidence of aggressive behaviour at 0 ppm. There were no clinical signs of toxicity. Food consumption and bodyweight increase were reduced during the treatment period at 5200 ppm. There were no treatment-related abnormalities observed post-mortem at termination. The LD50 value of the test substance to the Bobwhite quail was in excess of 5200 ppm. The no observed effect level in this study was 2600 ppm.

Reference 6

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

06 Feb 1992 to 14 Feb 1992

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)

Deviations:

not applicable

GLP compliance:

yes

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

5 d

Dose descriptor:

LD50

Effect level:

> 5 200 other: ppm

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

5 d

Dose descriptor:

NOEL

Effect level:

325 other: ppm

Conc. / dose based on:

test mat.

Basis for effect:

body weight

There were no mortalities although one bird at 650 ppm was replaced during the pre-treatment period. This bird was small in size possibly as a result of aggressive behaviour from other birds in the pen. No clinical signs of toxicity were observed during the study and excretal consistency remained normal. Bodyweight increase during the treatment period was reduced at 650 ppm and above with the most noticeable effect at 5200 ppm. Food consumption was reduced during the treatment period at 1300 ppm and above. Again, this was most pronounced at 5200 ppm. No abnormalities were detected in any bird examined post-mortem at termination. It was not possible to determine an LD50 value from the data. This must be in excess of 5200 ppm, the maximum dose level used. The no-effect level was found to be 325 ppm. There were no mortalities nor signs of intoxication in any treatment group. Body weight was affected at 650 ppm and above, feed consumption was reduced at 1300 ppm and above. Based on these findings, the LD50 was > 5200 ppm and the NOEL was determined to be 325 ppm.

 

Analytical results:

Group	Nominal inclusion (ppm)	Mean analysed concentration (ppm)	Relative mean error (%)
Control	0	ND	-
3	163	163	0
4	325	311	-4.3
5	650	648	-0.3
6	1300	1220	-6.2
7	2600	2540	-2.3
8	5200	5010	-3.7

 

Validity criteria fulfilled:

yes

Conclusions:

There were no mortalities nor signs of intoxication in any treatment group. Body weight was affected at 650 ppm and above, feed consumption was reduced at 1300 ppm and above. LC50 > 5200 ppm; NOEC = 325 ppm.

Executive summary:

A study was performed to determine the subacute dietary toxicity (LC50) of the test substance to the Mallard duck. The method followed was based on that given in the United States Environmental Protection Agency (EPA) Pesticide Assessment Guidelines, Subdivision E, Hazard Evaluation, Wildlife and Aquatic Organisms, Series 71 - Avian and mammalian testing, § 71-2 Avian dietary LC50 test, dated October 1982 and draft revised guidelines, dated March 1988. Groups of 10 young Mallard ducklings were offered diet containing 163, 325, 650, 1300, 2600 and 5200 ppm of test substance. Two similar sized control groups were offered basal diet only. Test diets were introduced when the birds were 8 days old and were offered to the birds for 5 days. Test diets were then replaced with basal diet and the birds observed for a further 3 days. Observations, including clinical signs, bodyweight and food consumption were made during the study. The birds were then killed and ten control birds together with ten from the highest surviving group subjected to a gross post-mortem examination. There were no mortalities although one bird at 650 ppm was replaced during the pre-treatment period. This bird was small in size possibly as a result of aggressive behaviour from other birds in the pen. No clinical signs of toxicity were observed during the study and excretal consistency remained normal. Bodyweight increase during the treatment period was reduced at 650 ppm and above with the most noticeable effect at 5200 ppm. Food consumption was reduced during the treatment period at 1300 ppm and above. Again, this was most pronounced at 5200 ppm. No abnormalities were detected in any bird examined post-mortem at termination. It was not possible to determine an LC50 value from the data. This must be in excess of 5200 ppm, the maximum dose level used. The no-effect level was found to be 325 ppm.

Reference 7

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

23 Mar 1995 to 12 Oct 1995

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

Test organisms (species):

Colinus virginianus

Duration (if not single dose):

21 wk

Dose descriptor:

NOEL

Effect level:

31.6 mg/kg bw/day

Conc. / dose based on:

act. ingr.

Basis for effect:

reproductive parameters

Remarks on result:

other: no reproductive effects.

Remarks:

Recalculated value in agreement with the RMS statement. See 'Any other information on results incl. tables'.

ANALYSIS OF AMENDED FEED

For diet verification samples, replicates of four test concentrations (0 ppm, 30 ppm, 100 ppm and 300 ppm) were analysed. Average concentrations of the test item found in the treated diet verification samples were 27.68 ± 3.56 for the 30 ppm level; 100.99 ± 12.22 for the 100 ppm level and 293.35 ± 20.41 for the 300 ppm level. No quantifiable CGA215944 residues were found in the control samples. Results of stability tests gave no indication for substance loss during storage. Study results are therefore based on nominal concentrations.

MORTALITY AND SYMPTOMS OF TOXICITY

There were no treatment-related mortalities at any of the concentrations tested. Four mortalities in the control group (week 14: female with probably broken neck and extensive bruising; emaciated male, where its pair mate was euthanised; week 17: female with severe foot abrasions) and one in the 300 ppm group (week 20: female with severe leg and toe abrasions) can be considered incidental. Post-mortem examinations on all of the mortalities yielded no treatment-related findings. All surviving adults were subjected to a post-mortem examination following adult termination. There were no apparent treatment related findings at the 30, 100 and 300 ppm as concentrations. No overt signs of toxicity were observed in any of the treatment groups during the course of the test. Incidental clinical observations normally associated with cage wear or interactions among cage mates (e.g., foot lesions and abrasions) were noted in the groups. Anti-pecking devices were affixed to some birds in order to reduce injuries due to pecking. One female, in Cage 16, had a herniated uterus. Except for these incidental findings, all birds appeared normal throughout the test.

ADULT BODY WEIGHT AND FOOD CONSUMPTION

There were no statistically significant differences between the control and test treatments for feed consumption or body weights following the administration of treated feed. The data set for adult female body weight was heterogeneous, necessitating the use of the Kruskal-Wallis test, which detected no significant mean differences among groups.

REPRODUCTION

In summary, no significant differences between the control and test treatments were detected for the observed reproductive parameters. The data sets for No. eggs cracked/No. eggs laid, for No. fertile eggs/No. eggs set, for No. viable embryos/No. fertile eggs and for No. hatchlings/No. fertile eggs departed from normality and required the use of the Kruskal-Wallis test, which showed no significant deviations in the test groups from the control group. One female in the 300 ppm produced only 1 fertile egg out of 35 examined eggs throughout the test period. It is concluded that the infertility was inherent in either the male or female and was not related to the test material in the diet, taking into account the respective values for the remaining birds in that group (on average 41.2 fertile eggs out of 44.5 examined eggs). Hence, the data for this pair were excluded from statistical analysis of reproduction parameters. ANOVA detected a significant mean difference in the number of 14 day survivors among groups. Tukey’s post-hoc test indicated the significant differences were between group 2 (30 ppm) and groups 1, 3 and 4. However, as the group 2 mean was greater than the control group mean, the difference was not treatment-related.

The reproductive endpoints are presented in the table below.

 

Summary of effects on reproductive parameters in bobwhite quail

Reproductive parameters	Concentrations (mg active substance/kg feed)
	Control	30	100	300
Number of laying pairs	16	19	19	17
Mean eggs laid	47.5 ± 10.8	49.1 ± 12.9	51.4 ± 11.2	49.9 ± 9.3
Mean eggs set	47.5 ± 10.8	44.2 ± 12.2	46.7 ± 10.9	44.5 ± 8.7
Eggshell thickness (mm ± SD)	0.218 ± 0.017	0.224 ± 0.023	0.216 ± 0.014	0.218 ± 0.013
Mean eggs cracked/eggs laid	0.005 ± 0.010	0.008 ± 0.017	0.002 ± 0.010	0.010 ± 0.013
Mean fertile eggs/eggs set	0.969 ± 0.048	0.921 ± 0.152	0.953 ± 0.092	0.928 ± 0.110
Mean viable embryos/fertile eggs	0.992 ± 0.013	0.972 ± 0.037	0.993 ± 0.012	0.938 ± 0.111
Mean hatchlings/viable embryos	0.969 ± 0.027	0.958 ± 0.073	0.974 ± 0.025	0.965 ± 0.069
Mean hatchling body weight (g ± sd)	7.7 ± 0.3	7.6 ± 0.4	7.4 ± 0.4	7.5 ± 0.3
Mean 14-day old survivors/ hatchlings	0.709 ± 0.096	0.822 ± 0.067	0.711 ± 0.104	0.655 ± 0.077
Mean 14-day old body weight (g ± sd)	24.3 ± 1.9	23.6 ± 2.3	21.0 ± 1.3	24.9 ± 1.6

 

There were no signs of toxicity and no significant differences in mortality and reproduction between the control and treatment groups. In addition, there were no apparent treatment related effects on body weight and feed consumption. Thus, the NOEL was determined to be 300 mg/kg.

RMS STATEMENT

Recalculation of concentration in food to daily dose was performed, based on a mean daily food consumption of 113.8 g and a mean body weight of 1079.6 g over the 21-week exposure in the NOEL treatment group receiving a diet with 300 mg as/kg. The resulting NOEL amounts to 31.6 mg as/kg bw/d.

Validity criteria fulfilled:

yes

Conclusions:

There were no signs of toxicity and no significant differences in mortality and reproduction between the control and treatment groups. In addition, there were no apparent treatment related effects on body weight and feed consumption. Thus, the NOEL was determined to be 300 mg as/kg.

Executive summary:

Seventy six male and seventy six female bobwhite quails (Colinus virginianus) were randomly distributed into the control group and three treatment groups. Each treatment and control group contained 19 pairs of birds, with one male and one female per pen. The adult birds were housed indoors in pens (50.7 x 26 x 25 cm high) constructed of epoxy-coated wire mesh with a slanted floor to facilitate egg collection. Test diets were prepared from a basemix. The basemix was prepared by thoroughly mixing requisite amounts of Purina Layena and the substance. The three test diets (30, 100 and 300 ppm) were prepared from the basemix on an as needed basis. Homogeneity samples (top right, top left, middle and bottom) were taken from the mixing vessel and verification of the test substance concentrations in the test diet was accomplished by analysing two samples per treatment group each time the feed was mixed. The test was initiated by providing the adult birds with treated food.

For all adult birds, daily observations were recorded throughout the test of signs of toxicity, mortality or abnormal behaviour. Adult body weights were measured at the beginning of acclimation, at the start of treated feed, at the end of week 10, and at adult termination. Body weights were not measured during egg laying because of the possible adverse effects occurring from handling of the test birds in that sensitive period. Feed consumption was determined weekly for each cage throughout the treated feed phase of the study by measuring the weight of the feeder at the beginning and end of each week, respectively, thereupon taking into account also added feed during the week and spillages captured in a catch pan under the feeding trays. At the conclusion of the exposure period, all adult birds were euthanised by carbon dioxide asphyxiation, subjected to a post-mortem examination, labelled and placed into frozen storage.

Eggs were collected daily from all cages, when available, and marked according to the cage of origin and date of collection. The eggs were then stored in a refrigerator (approx. 10 °C) until they were incubated. At weekly intervals, all eggs were removed from the refrigerator and logged onto the egg tracking forms. Per week, one egg from every second cage was collected, when available, for eggshell thickness measurement. The remaining eggs were candled with a candling lamp to detect eggshell cracks and other defects. Cracked or abnormal eggs were discarded. All eggs that were not cracked or used for eggshell thickness measurements were placed in a Nature Form Incubator (Model 1-14). Eggs were candled again between Day 10 and 12 of incubation to determine embryo viability and on the day eggs were transferred to the hatcher, usually on day 20, to determine embryo survival. On approx. day 20 of incubation, the eggs were placed in a Nature Form Hatcher (Model No. NOM 125) and allowed to hatch. Pedigree baskets constructed of stainless steel mesh were used to keep hatchlings segregated by cage of origin. All hatchlings, unhatched eggs and eggshells were removed from the hatcher on approx. day 22-24 of incubation. The body weight of each hatchling was then determined. Hatchlings were wing tagged for identification by cage of origin and then housed in brooding pens until 14 days of age. The hatchlings were fed untreated basal ration (Purina Startena). At 14 days of age the body weights of all surviving hatchlings were measured. The hatchlings were euthanised by carbon dioxide asphyxiation.

Upon completion of the test, statistical analyses were performed to determine significant differences between the control group and each treatment group in body weights (per individual), adult feed consumption and all of the reproductive parameters measured (per cage). The cages in which adult mortality occurred were not used in any statistical comparisons. Data sets were tested for normality using a Chi-Square test and for homogeneity of variance using Bartlett's test or Levene's test. Proportional data was Arcsine transformed and any data set departing from the normal distribution was transformed if the transformation resulted in a normal distribution. The data parameters were analysed by ANOVA with Tukey's post-hoc test for paired comparisons if the data exhibited homogeneity of variance and a normal distribution. If data were heterogeneous or non-normal, they were analysed with a Kruskal-Wallis test followed by Dunn's multiple pair comparisons test.

Analysis of amended feed: For diet verification samples, replicates of four test concentrations (0 ppm, 30 ppm, 100 ppm and 300 ppm) were analysed. Average concentrations of the test item found in the treated diet verification samples were 27.68 ± 3.56 for the 30 ppm level; 100.99 ± 12.22 for the 100 ppm level and 293.35 ± 20.41 for the 300 ppm level. No quantifiable CGA215944 residues were found in the control samples. Results of stability tests gave no indication for substance loss during storage. Study results are therefore based on nominal concentrations.

Mortality and symptoms of toxicity: There were no treatment-related mortalities at any of the concentrations tested. Four mortalities in the control group (week 14: female with probably broken neck and extensive bruising; emaciated male, where its pair mate was euthanised; week 17: female with severe foot abrasions) and one in the 300 ppm group (week 20: female with severe leg and toe abrasions) can be considered incidental. Post-mortem examinations on all of the mortalities yielded no treatment-related findings. All surviving adults were subjected to a post-mortem examination following adult termination. There were no apparent treatment related findings at the 30, 100 and 300 ppm as concentrations. No overt signs of toxicity were observed in any of the treatment groups during the course of the test. Incidental clinical observations normally associated with cage wear or interactions among cage mates (e.g., foot lesions and abrasions) were noted in the groups. Anti-pecking devices were affixed to some birds in order to reduce injuries due to pecking. One female, in Cage 16, had a herniated uterus. Except for these incidental findings, all birds appeared normal throughout the test.

Adult body weight and food consumption: There were no statistically significant differences between the control and test treatments for feed consumption or body weights following the administration of treated feed. The data set for adult female body weight was heterogeneous, necessitating the use of the Kruskal-Wallis test, which detected no significant mean differences among groups.

Reproduction: In summary, no significant differences between the control and test treatments were detected for the observed reproductive parameters. The data sets for No. eggs cracked/No. eggs laid, for No. fertile eggs/No. eggs set, for No. viable embryos/No. fertile eggs and for No. hatchlings/No. fertile eggs departed from normality and required the use of the Kruskal-Wallis test, which showed no significant deviations in the test groups from the control group. One female in the 300 ppm produced only 1 fertile egg out of 35 examined eggs throughout the test period. It is concluded that the infertility was inherent in either the male or female and was not related to the test material in the diet, taking into account the respective values for the remaining birds in that group (on average 41.2 fertile eggs out of 44.5 examined eggs). Hence, the data for this pair were excluded from statistical analysis of reproduction parameters. ANOVA detected a significant mean difference in the number of 14 day survivors among groups. Tukey’s post-hoc test indicated the significant differences were between group 2 (30 ppm) and groups 1, 3 and 4. However, as the group 2 mean was greater than the control group mean, the difference was not treatment-related.

There were no signs of toxicity and no significant differences in mortality and reproduction between the control and treatment groups. In addition, there were no apparent treatment related effects on body weight and feed consumption. Thus, the NOEL was determined to be 300 mg/kg.

Reference 8

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

26 Jun 1996 to 31 Dec 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-4 (Avian Reproduction Test)

Version / remarks:

1983

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

21 wk

Dose descriptor:

NOEL

Effect level:

300 mg/kg bw

Conc. / dose based on:

act. ingr.

Basis for effect:

reproductive parameters

ANALYSIS OF AMENDED FEED:

For diet verification samples, replicates of four test concentrations (0 ppm, 30 ppm, 100 ppm and 300 ppm) were analysed. Average concentrations of the test substance found in the treated diet verification samples were 32.77 ± 10.24 for the 30 ppm level; 113.43 ± 21.84 for the 100 ppm level and 362.71 ± 82.10 for the 300 ppm level. No quantifiable test substance residues were found in the control samples. Results are therefore based on nominal concentrations.

 

MORTALITY AND SYMPTOMS OF TOXICITY:

There were two adult duck mortalities during the treated feed portion of the study. A female in the control group died during the first week of treated feed. Two post mortem findings included slight emaciation and white spots on the heart. A female in the 100 ppm treated group died during the second week of treated feed after becoming entangled in the cage. Neither mortality was considered to be treatment-related. No abnormal behaviour was observed and the post-mortem observations identified no treatment related effects.

 

ADULT BODY WEIGHT AND FOOD CONSUMPTION:

There were no statistically significant differences between the control and test treatments for feed consumption or body weights following the administration of treated feed.

 

REPRODUCTION:

A significant difference was detected between the 100 mg as/kg treatment group and the control group in one reproductive parameter, survivor weights. Because there was no significant difference detected between the higher dose rate of 300 mg as/kg treatment group and the control, no trend was demonstrated and the effect is not considered to be treatment-related. The reproductive endpoints are presented in the table below.

Summary of effects on reproductive parameters in mallard duck

Reproductive parameters	Test substance concentration (mg/kg feed)
	Control	30	100	300
Number of laying pairs	15	16	15	16
Mean eggs laid	60.4 ± 9.8	52.2 ± 14.3	60.1 ± 7.9	56.3 ± 15.3
Mean eggs set	53.1 ± 11.1	45.8 ± 13.4	53.7 ± 8.1	46.3 ± 14.0
Eggshell thickness (mm ± SD)	0.360 ± 0.018	0.354 ± 0.018	0.360 ± 0.015	0.360 ± 0.013
Mean eggs cracked/eggs laid	0.053 ± 0.066	0.038 ± 0.060	0.028 ± 0.041	0.089 ± 0.120
Mean fertile eggs/eggs set	0.821 ± 0.264	0.903 ± 0.0.090	0.942 ± 0.050	0.851 ± 0.235
Mean viable embryos/fertile eggs	0.860 ± 0.113	0.833 ± 0.102	0.843 ± 0.098	0.854 ± 0.136
Mean hatchlings/viable embryos	0.573 ± 0.274	0.554 ± 0.227	0.537 ± 0.276	0.546 ± 0.219
Mean hatchling body weight (g ± sd)	34.7 ± 2.5	33.3 ± 1.7	33.8 ± 2.6	33.0 ± 3.1
Mean 14-day old survivors/hatchlings	0.843 ± 0.252	0.861 ± 0.080	0.831 ± 0.086	0.792 ± 0.133
Mean 14-day old body weight (g ± sd)	248.6 ± 27.1	233.0 ± 29.1	221.1 ± 22.0	251.8 ± 21.4

The long-term reproduction NOEL for the test substance to mallard (Anas platyrhynchos) was 300 mg as/kg, the highest concentration tested.

Validity criteria fulfilled:

yes

Conclusions:

The long-term reproduction NOEL for test substance to mallard duck (Anas platyrhynchos) was 300 mg as/kg, the highest concentration tested.

Executive summary:

Sixty four male and sixty four female mallard ducks (Anas platyrhynchos) were randomly distributed into the control group and three treatment groups. Each treatment and control group contained 16 pairs of birds, with one male and one female per pen. The adult birds were housed indoors in pens (76 × 83 × 44 cm high) constructed of polycarbonate-coated, galvanised, welded-wire mesh with a slanted floor to facilitate egg collection. The amended diet was prepared by directly adding the test substance to basal diet that was then mixed to ensure homogeneity using a large bulk mixer; in order to ensure freshness the procedure was repeated every three weeks throughout the exposure period. Four homogeneity samples (top right, top left, middle and bottom) were taken from the high and low treatment levels of one batch of diet prepared during the study. Two representative diet verification samples were taken from each batch of diet prepared during the study and were analysed for the test substance content. The test was initiated by providing the adult birds with treated food. Eggs were collected daily from pens, when available and were removed to a refrigerator (12.6 °C) prior to incubation. The eggs were ordered by cage and the number of eggs per cage recorded. The eggs were candled for cracking or abnormalities and a random sample of each was removed for eggshell thickness determination. The remaining eggs in each weekly set were incubated at 37.3-37.8°C and 50-65 % relative humidity. The eggs were candled on days 14 and 22 (embryo development and embryo survival respectively), and on day 22 viable eggs were removed to the egg hatcher. The hatcher maintained conditions of 36.6-37.5 °C and 62- 67 % relative humidity. Dividers were placed into the hatching trays to separate hatchlings by parental pen; hatchlings were removed to brooders over a 28-hour period. The hatchlings were weighed on the day of hatch and at the end of the 14-day observation period. Post mortem examination was conducted on any adult birds that died during the study and all those birds which remained at the end of egg laying. The environmental conditions were monitored throughout the study. During acclimatisation, birds were observed daily, and birds exhibiting abnormal behaviour or physical injury were not used for the test. All birds were observed daily throughout the test for signs of toxicity or abnormal behaviour. Adult body weights were measured 4 times during the study: at start of acclimation, at start of treated feed, during the pre-egg laying period and at adult termination. Adult feed consumption was measured weekly for each pen throughout the test, by weight of the feeding vessel (this must be considered as an estimate, due to wastage by the birds). Chicks were observed daily for signs of toxicity or abnormal behaviour. All adult birds were subjected to a gross necropsy. This included birds found dead or euthanised during the study, and all surviving adults sacrificed at test termination. Upon completion of the test, statistical analyses were performed to determine significant differences between the control group and each treatment group in body weights, adult feed consumption and all of the reproductive parameters measured. Data sets were firstly tested for normality and homogeneity using Chi-square, Bartlett’s or Levene’s test as appropriate; proportional data were then Arcsine transformed. Each data set was then analysed with ANOVA and an appropriate pair-wise comparison test. All statistical tests were carried out using TOXSTAT 1994 software and were based upon standard sources (Finney, 1971; Williams, 1971 and Williams 1972).

Analysis of amended feed: For diet verification samples, replicates of four test concentrations (0 ppm, 30 ppm, 100 ppm and 300 ppm) were analysed. Average concentrations of the test substance found in the treated diet verification samples were 32.77 ± 10.24 for the 30 ppm level; 113.43 ± 21.84 for the 100 ppm level and 362.71 ± 82.10 for the 300 ppm level. No quantifiable test substance residues were found in the control samples. Results are therefore based on nominal concentrations.

Mortality and symptoms of toxicity:There were two adult duck mortalities during the treated feed portion of the study. A female in the control group died during the first week of treated feed. Two post mortem findings included slight emaciation and white spots on the heart. A female in the 100 ppm treated group died during the second week of treated feed after becoming entangled in the cage. Neither mortality was considered to be treatment-related. No abnormal behaviour was observed and the post-mortem observations identified no treatment related effects.

Adult body weight and food consumption: There were no statistically significant differences between the control and test treatments for feed consumption or body weights following the administration of treated feed.

Reproduction: A significant difference was detected between the 100 mg as/kg treatment group and the control group in one reproductive parameter, survivor weights. Because there was no significant difference detected between the higher dose rate of 300 mg as/kg treatment group and the control, no trend was demonstrated and the effect is not considered to be treatment-related.

The long-term reproduction NOEL for the test substance to mallard ducks (Anas platyrhynchos) was 300 mg as/kg, the highest concentration tested.

Reference 9

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

25 Jun 1995 to 5 Jan 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-4 (Avian Reproduction Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 206 (Avian Reproduction Test)

Deviations:

no

GLP compliance:

yes

Test organisms (species):

Anas platyrhynchos

Duration (if not single dose):

22 wk

Dose descriptor:

NOEL

Effect level:

300 mg/kg bw

Conc. / dose based on:

act. ingr.

Basis for effect:

reproductive parameters

ANALYSIS OF AMENDED FEED:

Mean measured concentrations were 29, 100 and 260 mg as/kg feed (88-100 % of nominal). Results are therefore based on nominal concentrations.

 

MORTALITY AND SYMPTOMS OF TOXICITY:

A single mortality was observed in the control treatment. No abnormal behaviour was observed and the post-mortem observations identified no treatment related effects and all the observations were within the normal limits for general appearance and tissue deposition.

 

ADULT BODY WEIGHT AND FOOD CONSUMPTION:

There were no statistically significant differences between the control and test treatments for either body weight or food consumption.

 

REPRODUCTION:

The reproductive endpoints are presented in the table below.

 

Summary of effects on reproductive parameters in mallard duck

Reproductive parameters	Test substance concentration (mg/kg feed)
	Control	30	100	300
Number of laying pairs	15	16	15	16
Total eggs laid	894	1004	955	964
Eggs/female	59.6	62.8	59.6	60.3
Number of eggs cracked	22	46	48	11
Eggs cracked/eggs laid	0.025 ± 0.032	0.049 ± 0.068	0.049 ± 0.061	0.013 ± 0.026
Eggs set in incubator	806	884	841	889
Eggshell thickness (mm ± SD)	0.347 ± 0.025	0.336 ± 0.027	0.343 ± 0.017	0.344 ± 0.019
Fertile eggs	759	787	764	841
Fertile eggs/eggs incubated	0.94 ± 0.04	0.88 ± 0.21	0.91 ± 0.10	0.94 ± 0.05
Viable embryos	707	702	655	746
Viable embryos/fertile eggs	0.93 ± 0.09	0.88 ± 0.13	0.86 ± 0.15	0.89 ± 0.12
Hatchlings	653	596	551	624
Hatchlings/viable embryos	0.92 ± 0.08	0.82 ± 0.16	0.84 ± 0.12	0.83 ± 0.13
Mean hatchling body weight (g ± SD)	37.3 ± 3.1	38.1 ± 3.3	36.0 ± 3.2	36.3 ± 2.8
14-day old survivors	650	588	547	619
14-day old survivors/ hatchlings	1.0 ± 0.01	0.99 ± 0.02	0.99 ± 0.02	0.99 ± 0.01
Mean 14-day old body weight (g±SD)	246 ± 48.3	244 ± 40.1	240 ± 48.1	234 ± 43.1

 

The long-term reproduction NOEL for the test substance to mallard (Anas platyrhynchos) was 300 mg as/kg, the highest concentration tested.

Validity criteria fulfilled:

yes

Conclusions:

The long-term reproduction NOEL for the test substance to mallard duck (Anas platyrhynchos) was 300 mg as/kg, the highest concentration tested.

Executive summary:

Sixty four male and sixty four female mallard ducks (Anas platyrhynchos) were randomly distributed into the control group and three treatment groups. Each treatment and control group contained 16 pairs of birds, with one male and one female per pen. The adult birds were housed indoors in pens (76 × 83 × 44 cm high) constructed of polycarbonate-coated, galvanised, welded-wire mesh with a slanted floor to facilitate egg collection. The amended diet was prepared by directly adding test substance to basal diet that was then mixed to ensure homogeneity using a Hobart mixer; in order to ensure freshness the procedure was repeated every three weeks throughout the exposure period. Six verification samples were removed from each treatment and control at three separate mixing events and were analysed for the test substance content. The test was initiated by providing the adult birds with treated food. Eggs were collected daily from pens, when available (first eggs were set during week 12), and were removed to a controlled environmental chamber (16±1°C, 64-65 % RH) for up to one week prior to incubation. The eggs were ordered by cage and the number of eggs per cage recorded. The eggs were candled for cracking or abnormalities and a random sample were removed for eggshell thickness determination. The remaining eggs in each weekly set were incubated at 37-38 °C and 52-55 % relative humidity. The eggs were candled on days 14 and 23 (embryo development and embryo survival respectively), and on day 23 viable eggs were removed to the egg hatcher. The hatcher was programmed to maintain 36-37°C and 72-77 % relative humidity and dividers were placed into the hatching trays to separate hatchlings by parental pen. Hatchlings were removed to brooders over a 48-hour period commencing on day 27 post incubation. The hatchlings were weighed on the day of hatch and at the end of the 14-day observation period. Post mortem examination was conducted on any adult birds that died during the study and all those birds which remained at the end of egg laying. The environmental conditions were monitored throughout the study. During acclimatisation, birds were observed daily, and birds exhibiting abnormal behaviour or physical injury were not used for the test. All birds were observed daily throughout the test for signs of toxicity or abnormal behaviour. Adult body weights were measured 8 times during the study: at start of acclimation, just prior to the experimental start date, at the end of weeks 2, 4, 6 and 8 of the pre-laying period, at the start of photo stimulation and at adult termination. Adult feed consumption was measured weekly for each pen throughout the test, by weight of the feeding vessel (this must be considered as an estimate, due to wastage by the birds). Chicks were observed daily for signs of toxicity or abnormal behaviour. All adult birds were subjected to a gross necropsy. This included birds found dead or euthanised during the study, and all surviving adults sacrificed at test termination. Upon completion of the test, statistical analyses were performed to determine significant differences between the control group and each treatment group in body weights, adult feed consumption and all of the reproductive parameters measured. Data sets were firstly tested for normality and homogeneity using Chi-square, Bartlett’s or Levene’s test as appropriate; proportional data were then Arcsine or Anscombe Arcsine transformed. Each data set was then analysed with ANOVA and an appropriate pair-wise comparison test (Dunnett’s or Bonferroni’s test or a non-parametric Kruskal-Wallis ANOVA). All statistical tests were carried out using TOXSTAT Version 3.4 (West Inc, 1995) software.

Analysis of amended feed: Mean measured concentrations were 29, 100 and 260 mg as/kg feed (88-100 % of nominal). Results are therefore based on nominal concentrations.

Mortality and symptoms of toxicity: A single mortality was observed in the control treatment. No abnormal behaviour was observed and the post-mortem observations identified no treatment related effects and all the observations were within the normal limits for general appearance and tissue deposition.

Adult body weight and food consumption: There were no statistically significant differences between the control and test treatments for either body weight or food consumption.

The long-term reproduction NOEL for the test substance to mallard duck (Anas platyrhynchos) was 300 mg as/kg, the highest concentration tested.

Description of key information

The toxicity to birds was determined in several studies:

- 14-day oral LD50 for bobwhite quail was >2000 mg/kg bw, EPA FIFRA 71-1, Hakin, 1993

- 14-day oral LD50 for Japanese quail was >2000 mg/kg bw, EPA FIFRA 71-1, Hadhazy, 1991

- 14-day oral LD50 for Mallard duck was >2000 mg/kg bw, EPA FIFRA 71-1, Hadhazy, 1990

- No LD50 could be determined for Mallard duck. The 50% emetic dose (ED50) was 82.5 mg/kg bw, and NOEL was 31.25 mg/kg bw, EPA FIFRA 71-1, Hakin, 1993

- 5-d dietary LD50 for Bobwhite quail was > 5200 ppm. The NOEL was 2600 ppm. EPA FIFRA 71-2, Hakin 1992

- 5-d dietary LD50 for Mallard duck was > 5200 ppm. The NOEL was 325 ppm. EPA FIFRA 71-2, Hakin 1992

- No effects observed in a 21-wk reproduction test with Bobwhite quail. The NOEL was 300 mg/kg bw, EPA FIFRA 71-4, Taliaferro, 1996

- No effects observed in a 21-wk reproduction test with Mallard duck. The NOEL was 300 mg/kg bw, EPA FIFRA 71-4, Taliaferro, 1997

- No effects observed in a 22-wk reproduction test with Mallard duck. The NOEL was 300 mg/kg bw, EPA FIFRA 71-4, Stafford 2004

Information on this endpoint is not part of Annex VIII data requirements; all included studies are marked as supporting information and considered in the CSA.