reaction mass of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-2H -isothiazol-3-one

Toxicity to birds

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: US EPA FIFRA Guideline 71-2

Deviations:

no

GLP compliance:

yes

Remarks:

The concentration of active ingredient in the test material as stated in the Certificate of Analysis was not conducted under a GLP program

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Rohm and Haas, Batch No. 24088

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stable at room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: 458.7 g of CMIT/MIT was mixed into 12541 g Purina® Game Bird Startena
- Final dilution of a dissolved solid, stock liquid or gel: 5000 ppm CMIT/MIT, nominal

OTHER SPECIFICS: Purity of test material was 14.17%

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

no

Details on preparation and analysis of diet:

DIET PREPARATION
- Description and nutrient analysis of basal diet provided in study report: No
- Preparation of doses: Mixing of a blend of CMIT/MIT and Purina Game Bird Startena with an appropriate amount of hammer-milled stock diet

HOMOGENEITY AND STABILITY OF TEST MATERIAL IN DIET
- When and at what dose levels were samples of treated food analyzed for stability and concentration during the study: All dose levels, initial and after 6 days of refrigeration
- Nominal concentration (mg/kg feed): 312, 625, 1250, 2500 and 5000 ppm CMIT/MIT
- Concentration analysed (mg/kg feed): 102, 226, 616, 1450 and 3536 ppm CMIT/MIT
- % of nominal: 33-70%
- Mixing procedure adequate and variance between nominal and actual dosage acceptable (yes/no): no data

Test organisms (species):

Colinus virginianus

Details on test organisms:

TEST ORGANISM
- Common name: Bobwhite quail
- Source: Oak Ridge Game Farm, Gravette, Arkansas, USA.
- Age at test initiation (mean and range, SD): 11 days
- Weight at test initiation (mean and range, SD): 20-22 g
- Sexes used: Indeterminate sex
- Disease free: yes
- Kept according to standard practices: Not described

Limit test:

no

Total exposure duration (if not single dose):

5 d

Post exposure observation period:

3 days

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

10 animals of indeterminate sex per dose

Control animals:

yes, plain diet

Nominal and measured doses / concentrations:

Nominal: 312, 625, 1250, 2500 and 5000 ppm CMIT/MIT.
Measured: 102, 226, 616, 1450 and 3536 ppm CMIT/MIT.

Details on test conditions:

ACCLIMATION
- Acclimation period: 11 days
- Acclimation conditions (same as test or not): Yes
- Feeding: Purina Game Bird Startena
- Health (any disease or mortality observed): All birds were healthy and active at the end of the quarantine period.
- Fasting period before study: Not described

PEN SIZE AND CONSTRUCTION MATERIALS
- Description: Indoor holding pens
- Floor covering: Wire pens maintained over a concrete slab
- Compliant to good husbandry practices: Not described
- Suitable to avoid crowding stress: Not described
- Caging: Group

NO. OF BIRDS PER REPLICATE
- For negative control: 10
- For treated: 10

NO. OF REPLICATES PER GROUP
- For negative control: 5
- For treated: 1

TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 33.3 to 44.4 °C
- Relative humidity (%): 33 to 57 %
- Photoperiod: Fluorescent lighting, 24 hours a day.
- Ventilation: Not described
- Shielding: Not described

Details on examinations and observations:

MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: Daily

BODY WEIGHT
- Time schedule for examinations: Day 1, 5 and 8

FOOD CONSUMPTION (if feeding study)
- Time schedule for examinations: Last day of quarantine period, daily during 5-day test period and during the 3-day recovery period

Reference substance (positive control):

no

Key result

Duration (if not single dose):

8 d

Dose descriptor:

LC50

Effect level:

3 532.46 mg/kg diet

Conc. / dose based on:

act. ingr.

Basis for effect:

mortality

Remarks on result:

other: 2852-4385

Mortality and sub-lethal effects:

MORTALITY
- Results: 30% mortality in the group exposed to 3532.46 ppm CMIT/MIT/day after 5 days, increased to 50% after 8 days.

CLINICAL SIGNS
- Results: Lethargy and ataxia
- Remarks: Reversible in all surviving birds

BODY WEIGHT
- Results: Reduced food consumption, reversible in surviving birds.
- Remarks: All the control and test birds were noted as being smaller in size than normal attributed to reduced palatability of hammer-milled feed and consequently reduced feeding.

FOOD CONSUMPTION (if feeding study)
- Results: No significant effects

PATHOLOGY
- Results: Gross pathology of decedents and survivors revealed no abnormal pathological findings.

Further details on results:

The TS in the feed stored in the refrigerator for up to 5 days is stable; however, the TS partially degrades at elevated temperatures in the animal room. The analyses displayed that the lower the initial test concentration, the greater the degradation.

Reported statistics and error estimates:

LD50 calculation: Litchfield, J.T., Jr and Wilcoxon, F. “A Simplified Method of Evaluating Dose-Effect Experiments” Journal of Pharmacology and Experimental Therapeutics 96 (2):99 (1949).

Validity criteria fulfilled:

yes

Conclusions:

The LC50 for the short term dietary study with bobwhite quail was determined to be 3532.46 ppm CMIT/MIT. The NOEL was 226 ppm a.i. in feed/d based on weight and food consumption.

Executive summary:

US EPA FIFRA Guideline 71-2, An 8-day dietary toxicity study in bobwhite quail with analytical confirmation of dietary concentrations of CMIT/MIT. The TS in the feed stored in the refrigerator for up to 5 days is stable; however, the TS partially degrades at elevated temperatures in the animal room. The analyses displayed that the lower the initial test concentration, the greater the degradation. The LC50 for the short term dietary study with bobwhite quail was determined to be 3532.46 ppm CMIT/MIT mean measured since the feeds were given fresh daily. The NOEL was 226 ppm a.i. in feed/d based on weight and food consumption.

Reference 2

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: US EPA FIFRA Guideline 71-2

Deviations:

no

GLP compliance:

yes

Remarks:

The concentration of active ingredient in the test material as stated in the Certificate of Analysis was not conducted under a GLP program

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Rohm and Haas, Batch No. 24088

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stable at room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: 458.7 g of CMIT/MIT was mixed into 1,2541 g Purina Game Bird Startena
- Final dilution of a dissolved solid, stock liquid or gel: 5000 ppm CMIT/MIT, nominal

OTHER SPECIFICS: Purity of test material was 14.17%

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

no

Details on preparation and analysis of diet:

DIET PREPARATION
- Description and nutrient analysis of basal diet provided in study report: No
- Preparation of doses: Mixing of a blend of CMIT/MIT and Purina Game Bird Startena with an appropriate amount of hammer-milled stock diet

HOMOGENEITY AND STABILITY OF TEST MATERIAL IN DIET
- How often was homogeneity and stability tested: No data
- When and at what dose levels were samples of treated food analyzed for stability and concentration during the study: Initial and after 6 days of refrigeration
- Nominal concentration (mg/kg feed): 312, 625, 1250, 2500 and 5000 ppm CMIT/MIT
- Concentration analysed (mg/kg feed): 102, 226, 616, 1450 and 3536 ppm CMIT/MIT
- % of nominal: 33 to 70%
- Mixing procedure adequate and variance between nominal and actual dosage acceptable (yes/no): Not described

Test organisms (species):

Anas platyrhynchos

Details on test organisms:

TEST ORGANISM
- Common name: Mallard duck
- Source: Whistling Wings, Inc., Hanover, Illinois, USA
- Age at test initiation (mean and range, SD): 5 days
- Weight at test initiation (mean and range, SD): 47-50 g
- Sexes used: Indeterminate sex
- Disease free: Yes
- Kept according to standard practices: Not described

Limit test:

no

Total exposure duration (if not single dose):

5 d

Post exposure observation period:

3 days

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

10 animals of indetermine sex per dose

Control animals:

yes, plain diet

Nominal and measured doses / concentrations:

Nominal: 312, 625, 1250, 2500 and 5000 ppm CMIT/MIT
Measured: 102, 226, 616, 1450 and 3536 ppm CMIT/MIT

Details on test conditions:

ACCLIMATION
- Acclimation period: 2 days
- Acclimation conditions (same as test or not): Yes
- Feeding: Purina® Game Bird Startena
- Health (any disease or mortality observed): No
- Fasting period before study: Not described

PEN SIZE AND CONSTRUCTION MATERIALS
- Description: Indoor holding pens
- Floor covering: Wire pens maintained over a concrete slab
- Compliant to good husbandry practices: Not described
- Suitable to avoid crowding stress: Not described
- Caging: Group

NO. OF BIRDS PER REPLICATE
- For negative control: 10
- For treated: 10

NO. OF REPLICATES PER GROUP
- For negative control: 5
- For treated: 1

TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 21.6 to 26.7 °C
- Relative humidity (%): 75 to 91 %
- Photoperiod: Fluorescent lighting, 24 hours a day
- Ventilation: Not described
- Shielding: Not described

Details on examinations and observations:

MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: Daily

BODY WEIGHT
- Time schedule for examinations: Day 1, 5 and 8

FOOD CONSUMPTION (if feeding study)
- Time schedule for examinations: Last day of quarantine period, daily during 5-day test period and during the 3-day recovery period.

Reference substance (positive control):

no

Key result

Duration (if not single dose):

8 d

Dose descriptor:

NOEL

Effect level:

226 mg/kg diet

Conc. / dose based on:

act. ingr.

Basis for effect:

mortality

Key result

Duration (if not single dose):

8 d

Dose descriptor:

LC50

Effect level:

945 mg/kg diet

Conc. / dose based on:

act. ingr.

Basis for effect:

mortality

Mortality and sub-lethal effects:

MORTALITY
- Results: At the end of exposure period (5 days): 30, 70 and 100% mortality in the groups exposed to 616, 1450 and 3536 ppm CMIT/MIT, respectively. No increased mortality after 8 days.

CLINICAL SIGNS
- Results: Lethargy and anorexia
- Remarks: Reversible in all surviving birds

BODY WEIGHT
- Results: No significant effects
- Remarks: All the control birds were noted as being smaller in size than normal attributed to reduced palatability of hammer-milled feed and consequently reduced feeding.

FOOD CONSUMPTION (if feeding study)
- Results: No significant effects after 8 days
- Remarks: 100% mortality in the group dosed 5000 ppm CMIT/MIT (nominal)

PATHOLOGY
- Results: Gross pathology of decedents and survivors revealed no abnormal pathological findings.

Further details on results:

The TS in the feed stored in the refrigerator for up to 5 days is stable; however, the TS partially degrades at elevated temperatures in the animal room. The analyses displayed that the lower the initial test concentration, the greater the degradation.

Reported statistics and error estimates:

LD50 calculation: Litchfield, J.T., Jr and Wilcoxon, F. “A Simplified Method of Evaluating Dose-Effect Experiments” Journal of Pharmacology and Experimental Therapeutics 96 (2):99 (1949).

Validity criteria fulfilled:

yes

Conclusions:

The LC50 was determined to be 945 ppm CMIT/MIT (mean measured). The NOEL for the short term dietary study with mallard duck was 226 ppm a.i..

Executive summary:

US EPA FIFRA Guideline 71-2, An 8-day dietary toxicity study in mallard ducklings with analytical confirmation of dietary concentrations of CMIT/MIT. The TS in the feed stored in the refrigerator for up to 5 days is stable; however, the TS partially degrades at elevated temperatures in the animal room. The analyses displayed that the lower the initial test concentration, the greater the degradation. The LC50was determined to be 945ppm CMIT/MITmean measured since the feeds were given fresh daily. The NOEL for the short term dietary study with mallard duck was 226 ppm a.i. in feed/d.

Reference 3

Endpoint:

short-term toxicity to birds: acute oral toxicity test

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Rohm and Haas, Batch No. 24088

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stable at room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Undiluted, single oral dose via gelatin capsules

OTHER SPECIFICS: Purity of test material was 14.17%

Dose method:

capsule

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and analysis of diet:

DIET PREPARATION
- Description and nutrient analysis of basal diet provided in study report: No
- Preparation of doses: Volumetrically measured doses
- Type and function of solvent/vehicle: Gelatin capsule

Test organisms (species):

Colinus virginianus

Details on test organisms:

TEST ORGANISM
- Common name: Bobwhite quail
- Source: Oak Ridge Game Farm, Gravette, Arkansas, USA
- Age at test initiation (mean and range, SD): 31-weeks old
- Weight at test initiation (mean and range, SD): Not described
- Sexes used: male/female
- Cultural background: Not described
- Disease free: All birds were healthy and active prior to test initiation
- Kept according to standard practices: Not described
- Breeding population (e.g. reproductive history): Not described

Limit test:

yes

Total exposure duration (if not single dose):

21 d

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

5

Control animals:

yes, concurrent vehicle

Nominal and measured doses / concentrations:

Single oral dose in gelatin capsule: 21.5, 46.4, 68.1, 100, 147 mg CMIT/MIT / kg body weight

Details on test conditions:

ACCLIMATION
- Acclimation period: 15 days
- Acclimation conditions (same as test or not): Yes
- Feeding: Purina Duck Grower W/O
- Health (any disease or mortality observed): All birds were healthy and active prior to test initiation
- Fasting period before study: No

PEN SIZE AND CONSTRUCTION MATERIALS
- Description: Indoor holding pens
- Floor covering: Wire pens maintained over steel racks
- Compliant to good husbandry practices: Not described
- Suitable to avoid crowding stress: Not described
- Caging: Group

NO. OF BIRDS PER REPLICATE
- For negative control: n/a
- For vehicle control: 10
- For treated: 10

NO. OF REPLICATES PER GROUP
- For negative control: n/a
- For vehicle control: 1
- For treated: 1

TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 22.2 to 31.1 °C
- Brooder temperature: Not described
- Room temperature: Not described
- Relative humidity (%): 58 to 87 %
- Photoperiod: 10 hours light
- Ventilation: Not described
- Shielding: No

RANGE FINDING STUDY
- Test concentrations: 21.5, 46.4, 68.1, 100, and 147 mg CMIT/MIT/kg body weight
- Test conditions: Same as final test
- Results used to determine the conditions for the definitive study: 0% mortality at 21.5 mg CMIT/MIT/kg body weight, 100% mortality at 100 and 147 mg CMIT/MIT/kg body weight

Details on examinations and observations:

MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: Daily

BODY WEIGHT
- Time schedule for examinations: Days 1, 3, 7, 14 and 21

FOOD CONSUMPTION (if feeding study)
- Time schedule for examinations: Days 3, 7, 14 and 21.

PATHOLOGY
- Dose groups that were examined: Day 21 or time of death
- Remarks: Decedents and survivors were subjected to a gross pathological examination

Details on reproductive parameters:

n/a

Reference substance (positive control):

no

Key result

Duration (if not single dose):

21 d

Dose descriptor:

LD50

Effect level:

64.5 mg/kg bw

Conc. / dose based on:

act. ingr.

Basis for effect:

mortality

Remarks on result:

other: 51.6-80.6

Duration (if not single dose):

21 d

Dose descriptor:

NOEL

Remarks on result:

not determinable

Mortality and sub-lethal effects:

MORTALITY
- Results: >=90% mortality after day 1 in groups treated with 100 and 147 mg CMIT/ MIT / kg bw. 30% mortality on day 1, increasing to 60% mortality up to day 21 in the group treated with 68.1 mg CMIT/ MIT / kg bw. 10% mortality in the group treated with 46.4 mg CMIT/ MIT / kg bw after day 13.

CLINICAL SIGNS
- Results: Signs of toxicity (chalky diarrhea, lethargy, anorexia, dyspnea, and tachypnea)
- Remarks: Reversibility of all clinical signs was achieved in survivors by Day 19.

BODY WEIGHT
- Results: Body weight loss after day 3 for concentrations over 46.4 mg CMIT/ MIT / kg bw
- Remarks: No survivors after day 3 in groups treated with 100 and 147 mg CMIT/ MIT / kg bw

FOOD CONSUMPTION (if feeding study)
- Results: No significant effects

PATHOLOGY
- Results: Gross pathology of decedents revealed fluid-filled, hemorrhagic crops and hemorrhagic lungs.
- Remarks: Gross pathology of survivors revealed no abnormal pathological findings.

Reported statistics and error estimates:

LD50 calculation: Litchfield, J.T., Jr and Wilcoxon, F. “A Simplified Method of Evaluating Dose-Effect Experiments” Journal of Pharmacology and Experimental Therapeutics 96 (2):99 (1949).
Body weights: Texas Instruments 95 Programmable Calculator, One-Way Analysis of Variance Program, Texas Instruments, Inc., Dallas, Texas, USA.

Validity criteria fulfilled:

yes

Conclusions:

21d LD50 = 64.5 mg CMIT/MIT/kg body weight. A no-observed-effect-level (NOEL) was not achieved in this study.

Executive summary:

US EPA FIFRA Guideline 71-1, An acute oral LD50 study in Bobwhite quail with TS administered via gelatin capsule. All birds were observed on the day of dosing and for 20 additional days thereafter. Twenty-seven birds died during the study. Signs of clinical toxicity were reversed in all surviving birds.  No gross pathological findings were noted in surviving birds.  The 21d LD50 was determined to be 64.5 mg CMIT/MIT/kg body weight. A no-observed-effect-level (NOEL) was not achieved in this study.

Description of key information

Results from toxicity studies on mallard duck (Anas platyrhynchos) and bobwhite quail (Colinus virginianus) demonstrate that C(M)IT/MIT exhibits slight to moderate toxicity to birds. The acute oral LD50for bobwhite quail is 460.72 mg/kg bw (equivalent to 64.5 mg a.i./kg bw) and the LD0 is 153.57 mg/kg bw (equivalent to 21.5 mg a.i./kg bw) . The short term dietary LC50 for mallard duck is 945 ppm a.i. and bobwhite quail is 3536 ppm a.i. . The NOEL and LC0 for the short-term dietary study with mallard duck was 226 ppm a.i. The LC0 for the short-term dietary study with bobwhite quail was and 1450 ppm a.i.