mancozeb (ISO); manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt  

Toxicity to birds

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1989-09-07 – 1990-04-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.2300 (Avian Reproduction Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Dose method:

homogenously mixed into feed (accounts for technical substances)

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Anas platyrhynchos

Details on test organisms:

- Source: Whistling Wings, Hanover, Illinois, USA
- Age: 25 weeks at test initiation
- Environmental conditions:
Temperature 19.8 ± 2.4°C
Photoperiod 8 hours light during first 8 weeks 17 hours light from beginning of week 9
Relative humidity 56%

Limit test:

no

Total exposure duration (if not single dose):

18 wk

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

one male and one female

Control animals:

yes, plain diet

Nominal and measured doses / concentrations:

Nominal: 0, 10, 50, 125 and 1000 ppm a.s.;
Measured: 9.65 ± 0.21 (96.5 ± 2.1%), 50.65 ± 1.28 (101.3 ± 2.5%), 122.25 ± 1.94 (97.8 ± 1.6%) and 956 ± 36 mg a.s./kg feed (95.6 ± 3.8%)

Details on test conditions:

ACCLIMATION
- Acclimation period: 8 weeks
- Acclimation conditions: same as test
- Feeding: up to 265 g/bird/day of the prepared diet
- Health: Pen-reared mallards (Anas platvrhvnchos) that were apparently healthy and phenotypically indistinguishable from wild birds, were purchased from Whistling Wings, Box 1, 113 Washington Street, Hanover, Illinois 61041. All birds were from the same hatch and were 25 weeks of age at the initiation of the test (first day of exposure to test diet). The birds were approaching their first breeding season and had not been used in previous testing. At test initiation all birds were examined for physical injuries and general health. Birds that did not appear healthy were discarded. Sex of the birds was determined by a Visual examination of the plumage.
- Fasting period before study: no

PEN SIZE AND CONSTRUCTION MATERIALS
- Description: Safeguard Products Inc. Model #5355 measuring approx. 75 X 90 X 45 cm high; pens constructed of galvanized wire grid and galvanized sheeting
- Floor covering: vinyl coated wire mesh
- Compliant to good husbandry practices: yes
- Suitable to avoid crowding stress: not specified
- Caging: batteries of brooding pens

NO. OF BIRDS PER REPLICATE
- For negative control: 1 male and 1 female
- For treated: 1 male and 1 female

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

TEST CONDITIONS
- Temperature incubator (breeding apparatus): 37.4 +/- 0.1 °C (SD)
- Relative humidity incubator (breeding apparatus) (%): 53
- Temperature (hatching compartment): 37.0 °C +/- 0.4 °C (SD)
- Average bulb temperature (hatching compartment): raised to 32.4 °C +/- 1.2 °C (SD)
- Relative humidity (hatching compartment) (%): ca. 70
- Photoperiod: for hatchlings 16 hours light and 8 hours darkness

Details on examinations and observations:

Adult body weight:
Individual body weight was measured at the experimental start, weeks 2, 4, 6, 8 and at termination of the study.

Adult feed consumption:
Feed consumption expressed as grans of feed per bird per day was examined by pen or each seven day period during the study.

Eggs laid of maximum laid:
The number of eggs laid per hen divided by the largest number of eggs laid by one hen.

Eggs cracked of eggs laid:
The number of eggs determined by candling to be cracked divided by the number of eggs laid, per pen.

Viable embryos of eggs set:
The number of viable embryos at day 14 candling was divided by the number of eggs set, per pen.

Live 3-week embryos of viable embryos:
The number of live embryos at day 24 candling was divided by the number of viable embryos.

Hatchlings of 3-week embryos:
The number of hatchlings removed from the hatcher was divided by the number of live 3 week embryos, per pen.

14-day old survivors of hatchlings:
The number of 14-day old survivors was divided by the number of hatchlings per week, by pen.

Hatchlings of eggs set:
The number of hatchlings was divided by the number of eggs set per week, by pen.

14-day old survivors of eggs set:
The number of 14-day old survivors was divided by the number of eggs set per week, by pen.

Hatchlings of maximum set:
The number of hatchlings per hen divided by the largest number of eggs set from any one hen. This transformation was used to convert the number of hatchlings to a percentile value equal to or less than 100.

14-day old survivors of maximum set:
The number of 14-day old survivors per pen divided by the largest number of eggs set.

Egg shell thickness:
The average egg shell thickness of randomly selected eggs per pen, was measured.

Offspring’s body weight:
The group body weights of hatchlings and 14-day old survivors was measured by parental groups.

Details on reproductive parameters:

please see above

Reference substance (positive control):

no

Key result

Duration (if not single dose):

18 wk

Dose descriptor:

NOEC

Effect level:

18.6 mg/kg bw/day

Conc. / dose based on:

act. ingr.

Basis for effect:

reproductive parameters

Remarks on result:

other: The 18 wk NOEC was recalculated by the Rapporteur Member State under to be 18.6 mg a.s./kg bw/d. For details please refer to the Renewal Assessment Report for Mancozeb prepared according the Commission Regulation (EU) No 1107/2009.

Mortality and sub-lethal effects:

There were no treatment-related mortalities at any Mancozeb concentration tested. One incidental mortality occurred in the 1000 mg a.s./kg feed treatment group in week 16 (necropsy showed severe egg yolk peritonitis and regressing ovary), but no mortalities occurred in the control group or the 10, 50 or 125 mg a.s./kg feed treatment groups. No overt signs of toxicity were observed at any concentration tested. Incidental clinical signs were noted in control and various treatment groups during the study. All surviving adults were necropsied at adult terminal sacrifice. At the 1000 mg a.s./kg feed test concentration there appeared to be a treatment related increase in the number of hens exhibiting lesions of the egg yolk peritonitis. There were no apparent treatment effects upon adult body weight at any test concentration.

Effects on reproduction:

There was no treatment-related effect upon the body weight of hatchlings or 14-day old survivors in at concentrations of 10, 50 or 125 mg a.s./kg feed. At 1000 mg a.s./kg feed, while not statistically significant, there appeared to be a reduction in the body weight of hatchlings and there was a statistically significant (p<0.05) reduction in the body weight of 14-day old survivors.

There were no apparent or significant treatment-related effects upon reproductive parameters at the test concentrations 10, 50 or 125 mg a.s./kg feed for any reproductive parameter. However at 1000 mg a.s./kg feed test concentration there was profound effect upon reproductive performance. A statistically significant (p<0.01) adverse effect was observed upon the number of eggs laid, viable embryos, live 3-week embryos, hatchability and the number of hatchlings and 14-day old survivors per hen. The onlyparameters not affected at 1000 mg a.s./kg feed were the percentage of cracked eggs and the survival of hatchlings of 14 days of age. While hens in the control, 10, 50 and 125 mg a.s./kg feed treatment groups averaged 20 or more offspring per hen, the average number of 14-day old survivors per hen in the 1000 mg a.s./kg feed treatment was 2. There were no apparent and significant treatment related effects upon eggshell thickness at any Mancozeb concentration.

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

The Dunnett’s multiple comparison procedure was used to determine statistically significant differences between parameters measured for control group and each of the treatment groups. Percentage data were analysed using Dunnett’s method following arcsine transformation. Significant differences are indicated in the tables with * p<0.05 and **p<0.01.

For tabels please refer to section on "Attached background material".

Validity criteria fulfilled:

yes

Conclusions:

There were no apparent treatment-related effects upon reproductive parameters at concentrations of 10, 5 and 125 ppm a.s. At 1000 ppm a.s. there was a marked effect upon reproductive performance, including decreases in the numbers of eggs laid, embryo viability, embryo survivability, hatchability, the numbers of hatchlings and 14 day old survivors per hen and body weight of offspring at hatch and after 14 days of growth and a related incidence of egg yolk peritonitis in an adult hen at necropsy. Based upon marked effects on reproductive performance at 1000 ppm a.s., the NOEL in this study for mallards exposed to Mancozeb was 125 ppm a.s. Converted to daily dietary dose the 18 wk NOEC is determined to be 18.6 mg a.s./kg bw/d.

Executive summary:

To evaluate the effects of dietary exposure of Mancozeb to the adult mallard (Anas platvrhvnchos) a one-generation study with adult mallard (Anas platvrhvnchos) was performed in accordance with the principles laid down in the guideline EPA OPPTS 850.2300 (Avian Reproduction Test). Effects on adult health, weight gain and feed consumption were assessed over an period of 18 weeks. In addition, the effects of adult exposure to Mancozeb on the number of eggs laid, normal development of eggs, viability of the embryos, percent hatchability, offspring survival and egg Shell thickness were evaluated. Four groups of adult mallards received diets containing Dithane M-45 at concentrations of 10, 50, 125 and 1000 mg a.s./kg feed (Mancozeb) for 18 weeks: 8 weeks prior to the start of egg production (pre-photostimulation period) and during 10 weeks of egg production. Each treatment group contained 16 replicates with one male and one female per pen. The fifth group of 16 replicates received untreated diet. All dietary concentrations were adjusted for test substance reported purity of 80.1% and are given in a.s. (Mancozeb). The study phases were: acclimation (8 weeks), pre-photostimulation (8 weeks), egg laying period (10 weeks) and post-adult sacrifice (final incubation, hatching and offspring rearing period; 6 weeks). During the acclimation period birds were given untreated basal diet and they were maintained under a photoperiod of 8 hours light. At beginning of test week 9 the photoperiod was increased to 17 hours light and 7 hours darkness. Effects on adult health, weight gain, feed consumption and reproduction were evaluated. Eggs were collected from each pen daily over the 10-week egg production period from the start of week 9 until the end of week 18. Egg shell thickness was measured, broken and cracked eggs were recorded and discarded. Eggs (except those for shell thickness measurements) were incubated for 24 days until fertile eggs were transferred into a hatcher. Effects on egg production, fertility and embryo survival (candling day 14 and 21) as well as hatchling health and survivability were examined. Adult birds which died during and all study and all adult birds after termination of the study were examined post-mortem. Diets for adult birds and their offspring were formulated to Wildlife International Ltd specifications by Agway Inc. Water and food were provided ad libitum throughout acclimation and the test. Hatchlings were housed at 20.9 ± 1.8°C with a photoperiod of 16 hours light/8 hours dark cycle. Adult test diets were prepared once each week. Six homogeneity samples from control and each test concentration were collected on day 0. Stability samples were collected on day 7 and 14. Verification samples of control and each test diet were taken immediately after mixing and stored on the study room for 7 days. On day 7, all samples were placed in a freezer and stored until shipped to the analytical laboratory of Rohm&Haas Company. Samples were taken for the first four weeks and then every fourth week thereafter. As a result, the homogeneity of Mancozeb concentrations varied between 1.6 — 3.8% between top. middle and bottom of the mixed diet. Averaged concentrations between top. middle and bottom were determined to be 9.65 ± 0.21 mg a.s/kg feed (96.5 ± 2.1%), 50.65 ± 1.28 mg a.s./kg feed (101.3 ± 2.5%). 122.25 ± 1.94 mg a.s./kg feed (97.8 ± 1.6%) and 956 ± 36 mg a.s./kg feed (95.6 ± 3.8%). Freshly analysed food had an average of 97.8% target concentration averaged between treatment groups, while food stored frozen for 30 days had an average of 94.0% target concentration averaged between treatment groups. After 7 or 14 days storage at ambient temperature Mancozeb concentrations were 85.5% and 72.2% of target concentrations averaged of treatment groups, respectively. The applied concentrations did not result in treatment-related mortality, overt signs of toxicity or effects upon adultbody weight or feed consumption. There were no apparent treatment-related effects upon reproductive parameters at concentrations of 10, 5 and 125 ppm a.s. At 1000 ppm a.s. there was a marked effect upon reproductive performance, including decreases in the numbers of eggs laid, embryo viability, embryo survivability, hatchability, the numbers of hatchlings and 14 day old survivors per hen and body weight of offspring at hatch and after 14 days of growth and a related incidence of egg yolk peritonitis in an adult hen at necropsy. Based upon marked effects on reproductive performance at 1000 ppm a.s., the NOEL in this study for mallards exposed to Mancozeb was 125 ppm a.s. Converted to the daily dietary dose the 18 wk NOEC is determined to be 18.6 mg a.s./kg bw/d.

Description of key information

In an one-generation study following the principles of EPA Guideline OPPTS 850.2300 the 18-week NOEC was determined to be 18.6 mg a.s./kg bw/d.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for birds:

18.6 mg/kg bw/day

Additional information

Key study

A One-Generation Reproduction Study with the Mallard (Anas platyrhynchos) Laboratory: Wildlife International Ltd. (Beavers et al. 1991 (Doc. No. 813-002, cross reference to Risk Assessment Report according to Regulation (EU) No 1107/2009: 8.1.1.3)

To evaluate the effects of dietary exposure of Mancozeb to the adult mallard (Anas platvrhvnchos) a one-generation study was performed in accordance with the principles laid down in the guideline EPA OPPTS 850.2300 (Avian Reproduction Test) (Beavers et al., 1991). Effects on adult health, weight gain and feed consumption were assessed over an period of 18 weeks. In addition, the effects of adult exposure to Mancozeb on the number of eggs laid, normal development of eggs, viability of the embryos, percent hatchability, offspring survival and egg Shell thickness were evaluated. Four groups of adult mallards received diets containing Dithane M-45 at concentrations of 10, 50, 125 and 1000 mg a.s./kg feed (Mancozeb) for 18 weeks: 8 weeks prior to the start of egg production (pre-photostimulation period) and during 10 weeks of egg production. Each treatment group contained 16 replicates with one male and one female per pen. The fifth group of 16 replicates received untreated diet. All dietary concentrations were adjusted for test substance reported purity of 80.1% and are given in a.s. (Mancozeb). The study phases were: acclimation (8 weeks), pre-photostimulation (8 weeks), egg laying period (10 weeks) and post-adult sacrifice (final incubation, hatching and offspring rearing period; 6 weeks). During the acclimation period birds were given untreated basal diet and they were maintained under a photoperiod of 8 hours light. At beginning of test week 9 the photoperiod was increased to 17 hours light and 7 hours darkness. Effects on adult health, weight gain, feed consumption and reproduction were evaluated. Eggs were collected from each pen daily over the 10-week egg production period from the start of week 9 until the end of week 18. Egg shell thickness was measured, broken and cracked eggs were recorded and discarded. Eggs (except those for shell thickness measurements) were incubated for 24 days until fertile eggs were transferred into a hatcher. Effects on egg production, fertility and embryo survival (candling day 14 and 21) as well as hatchling health and survivability were examined. Adult birds which died during and all study and all adult birds after termination of the study were examined post-mortem. Diets for adult birds and their offspring were formulated to the test facility's specifications by Agway Inc. Water and food were provided ad libitum throughout acclimation and the test. Hatchlings were housed at 20.9 ± 1.8 °C with a photoperiod of 16 hours light/8 hours dark cycle. Adult test diets were prepared once each week. Six homogeneity samples from control and each test concentration were collected on day 0. Stability samples were collected on day 7 and 14. Verification samples of control and each test diet were taken immediately after mixing and stored on the study room for 7 days. On day 7, all samples were placed in a freezer and stored until shipped to the analytical laboratory of Rohm&Haas Company. Samples were taken for the first four weeks and then every fourth week thereafter. As a result, the homogeneity of Mancozeb concentrations varied between 1.6 — 3.8 % between top. middle and bottom of the mixed diet. Averaged concentrations between top. middle and bottom were determined to be 9.65 ± 0.21 mg a.s/kg feed (96.5 ± 2.1 %), 50.65 ± 1.28 mg a.s./kg feed (101.3 ± 2.5 %). 122.25 ± 1.94 mg a.s./kg feed (97.8 ± 1.6 %) and 956 ± 36 mg a.s./kg feed (95.6 ± 3.8 %). Freshly analysed food had an average of 97.8% target concentration averaged between treatment groups, while food stored frozen for 30 days had an average of 94.0 % target concentration averaged between treatment groups. After 7 or 14 days storage at ambient temperature Mancozeb concentrations were 85.5% and 72.2% of target concentrations averaged of treatment groups, respectively. The applied concentrations did not result in treatment-related mortality, overt signs of toxicity or effects upon adultbody weight or feed consumption. There were no apparent treatment-related effects upon reproductive parameters at concentrations of 10, 5 and 125 ppm a.s. At 1000 ppm a.s. there was a marked effect upon reproductive performance, including decreases in the numbers of eggs laid, embryo viability, embryo survivability, hatchability, the numbers of hatchlings and 14 day old survivors per hen and body weight of offspring at hatch and after 14 days of growth and a related incidence of egg yolk peritonitis in an adult hen at necropsy. Based upon marked effects on reproductive performance at 1000 ppm a.s., the NOEL in this study for mallards exposed to Mancozeb was 125 ppm a.s. Converted to the daily dietary dose the 18 wk NOEC is determined to be 18.6 mg a.s./kg bw/d.