N-[3-(dimethylamino)propyl] C6-9 alkyl amides

Toxicity to birds

Administrative data

Endpoint:

short-term toxicity to birds: acute oral toxicity test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 18, 2009 to to May 04, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.2100 (Avian Acute Oral Toxicity Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

1044764-00-2

Molecular formula:

RCONHCH2CH2CH2N(CH3)2

Test material form:

liquid

Dose method:

other: the test substance in diluent was orally intubated directly into the crop or proventriculus

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and analysis of diet:

The test substance was dispersed in reverse osmosis water. The concentration of the test substance in the diluent was adjusted to provide a constant volume to body weight dosage for all treatment birds. The dosage was not adjusted to 100% active ingredient. Therefore, all dosages and the LD50 value are reported as milligrams of the test substance per kilogram of body weight. Nominal dosages used in this study were 0, 175, 292, 486, 810, and 1350 milligrams of test substance per kilogram of body weight (mg/kg).

Test organisms

Test organisms (species):

other: Northern bobwhite (Colinus virginianus)

Details on test organisms:

All northern bobwhite (Colinus virginianus) were approximately 28 weeks of age and appeared to be in good health at initiation of the test. Northern bobwhite ranged in weight from 176-228 grams at test initiation. The birds were obtained from M & M Quail Farm, Gillsville, GA 30543. Test birds were maintained separately by sex at Wildlife International, Ltd. under conditions that would not facilitate reproduction. All birds were from the same hatch, pen-reared and phenotypically indistinguishable from wild birds. Birds were assigned to five test groups and one control group. Each treatment or control group contained five males and five females. All test birds were acclimated to the study room and test caging for ten weeks prior to the initiation of the test.

Identification
Each pen was identified with a unique number, and groups of pens were identified by project number and dosage. Individual birds within each pen were identified by colored numbered leg bands.

Avian Feed and Water
Throughout acclimation and testing all test birds were fed a game bird ration formulated to specifications by Cargill Animal Nutrition Company, Shippensburg, PA 17257.
Water, from the town of Easton public water supply, and feed were provided ad libitum during acclimation and during the test, except during periods of fasting prior to testing.
Beginning two days after their arrival in the test facility, test birds were given soluble antibiotics in their drinking water for seven consecutive days. The birds received no form of antibiotic medication during the test.

Study design

Limit test:

no

Total exposure duration (if not single dose):

1 d

Post exposure observation period:

14 days

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

Each treatment or control group contained five males and five females.

Control animals:

yes, concurrent vehicle

Nominal and measured doses / concentrations:

Nominal Dosage: 0, 175, 292, 486, 810, and 1350 mg/kg body weight (mg/kg)

Details on test conditions:

Housing and Environmental Conditions

Test birds were housed indoors by dosage group in batteries of pens manufactured by GQF Manufacturing Co. (Model No. 0010). Each pen had floor space that measured approximately 78 X 51 cm. Floors were sloped so that ceiling height ranged from approximately 20 to 25 cm. External walls, ceilings and floors were constructed of wire mesh while side walls were constructed of galvanized sheeting. Each dosage group was assigned two pens. One pen contained five males and the other five females. Birds were assigned to pens by indiscriminate draw. Each group of birds was identified by pen number.

Birds were maintained at ambient room temperature. The average temperature for this study was 24.5 ± 0.4°C (SD) with an average relative humidity of 56 ± 13% (SD). The photoperiod (maintained by a time clock) was approximately eight hours of light per day/16 hours dark during acclimation and throughout the test. The light source was fluorescent lights, which closely approximate noonday sunlight. The birds were exposed to an average of approximately 246 lux of illumination. Housing and husbandry practices were based on guidelines established by the National Research Council.

Dosing

The birds were fasted for approximately 18 hours prior to dosing. At the experimental start, a single dose of the test substance in diluent was orally intubated directly into the crop or proventriculus of each bird using a stainless steel 14 gauge cannula. Each bird was individually weighed and dosed on the basis of milligrams of test substance per kilogram of body weight. The control birds received a corresponding volume of diluent only. All treatment and control birds received a constant dosage volume of four milliliters per kilogram of body weight.

Examinations

Details on examinations and observations:

During acclimation, all birds were observed daily. Birds exhibiting abnormal behavior or physical injury were not used. Following dosing, multiple observations were performed on Day 0 of the test, with particular attention being paid for signs of regurgitation. From test initiation until termination, all birds were observed at least twice daily. A record was maintained of all mortality, signs of toxicity, and abnormal behavior.

Reference substance (positive control):

no

Results and discussion

Effect levels

Key result

Effect level:

561 mg/kg bw

Conc. / dose based on:

test mat.

Remarks on result:

other: 95% confidence interval

Remarks:

441 to 716 mg/kg

Mortality and sub-lethal effects:

There were no mortalities in the control group. In addition, there were no mortalities in the 175 mg/kg treatment group. However, there was 10% (1 of 10) mortality at the 292 mg/kg dosage level, 30% (3 of 10) mortality at the 486 mg/kg dosage level, 80% (8 of 10) at the 810 mg/kg dosage level and 100% (10 of 10) mortality at the 1350 mg/kg dosage level.

All control birds were normal in appearance and behavior throughout the test with the exception of one female who was noted as limping on the left leg on Days 13 and 14 of the test. At the 175 mg/kg dosage level, signs of toxicity were first noted twenty-five minutes after dosing, when one bird was noted with a ruffled appearance. A total of six birds were noted with signs of toxicity on the day of dosing. Signs of toxicity noted at the 175 mg/kg dosage level were loss of coordination and ruffled appearance. All birds were normal in appearance and behavior from three hours and thirty minutes after dosing until test termination.

At the 292 mg/kg dosage level, signs of toxicity were first noted forty-one minutes after dosing, when nine birds were noted with signs of toxicity and one bird was found dead. Signs of toxicity noted at the 292 mg/kg dosage level were loss of coordination, shallow and rapid respiration, and ruffled appearance. All surviving birds were normal in appearance and behavior from two hours and forty-six minutes after dosing until test termination. At the 486 mg/kg dosage level, signs of toxicity were first noted twenty-three minutes after dosing, when all birds were noted with signs of toxicity. Within two hours of dosing three birds were found dead. Signs of toxicity noted at the 486 mg/kg dosage level were depression, wing droop, loss of coordination, prostrate posture, loss of righting reflex, shallow and rapid respiration, ruffled appearance, lower limb weakness, and lethargy. All surviving birds were normal in appearance and behavior from the morning of Day 1 until test termination. At the 810 mg/kg dosage level, signs of toxicity were first noted eight minutes after dosing, when all birds were noted with signs of toxicity. On the day of dosing, eight birds were found dead. Signs of toxicity noted at the 810 mg/kg dosage level were depression, loss of coordination, prostrate posture, loss of righting reflex, ruffled appearance, lower limb weakness, and lethargy. Both surviving birds were normal in appearance and behavior from the morning of Day 3 until test termination. At the 1350 mg/kg dosage level, signs of toxicity were first noted forty-two minutes after dosing, when nine birds were noted with signs of toxicity and one bird was found dead. On the day of dosing, a total of nine birds were found dead. The remaining bird was found dead on the morning of Day 2. Signs of toxicity noted at the 1350 mg/kg dosage level were depression, loss of coordination, prostrate posture, loss of righting reflex, ruffled appearance, lower limb weakness, lethargy, and minor muscle fasciculation.

Further details on results:

Body Weight and Feed Consumption

When compared to the control group, there were no treatment related effects on body weight among males or females at the 175 mg/kg dosage level. There was a treatment-related reduction in mean body weight gain among males and surviving females at the 292 mg/kg dosage level from Day 0 to Day 3. A loss of mean body weight was observed among surviving males and females at the 486 and 810 mg/kg dosage levels from Day 0 to Day 3. For all surviving birds mean body weight changes were comparable to or exceeded mean body weight changes for the control group from Day 4 to 7 and from Day 8 to 14. An overall loss of body weight was noted for the two surviving birds in the 810 mg/kg dosage level. Body weight effects among males and females in the 1350 mg/kg treatment group could not be evaluated due to total mortality at this level.
When compared to the control group, there were no apparent treatment-related effects on feed consumption among surviving males or females at the 175 and 292 mg/kg dosage levels or among surviving females at the 486 mg/kg dosage level. There were reductions in mean feed consumption for surviving males in the 486 mg/kg dosage level and among surviving males and females in the 810 mg/kg dosage level from Day 0 to Day 3 of the test that were considered to be treatment related. The surviving female in the 810 mg/kg dosage group continued to be noted with a reduction in feed consumption from Day 4 to Day 7. All other dosage groups feed consumption values from Day 4 to Day 7 were comparable to the control and all dosage groups feed consumption values from Day 8 to Day 14 were comparable to the control. Feed consumption among males and females in the 1350 mg/kg treatment group could not be determined due to total mortality in this group.

Reported statistics and error estimates:

The slope of the dose response curve was 5.516, with a Chi-square value of 0.704. The no-mortality level was 175 mg/kg.
Mortality data were analyzed using the computer program of C.E. Stephan. The program was designed to calculate the LD50 value and the 95% confidence interval by probit analysis, moving average method or the nonlinear interpolation method (5,6, 7). In this study the LD50 value was determined by probit analysis. No statistical analyses were applied to separate mean responses among treatment groups for the endpoints of food consumption and body weight.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the acute oral LD50 for northern bobwhite quail exposed to the test substance as a single oral dose was calculated to be 561 mg/kg bw, with a 95% confidence interval of 441 to 716 mg/kg bw.

Executive summary:

A study was conducted to determine the acute toxicity of the test substance administered to northern bobwhite quail as a single oral dose and to determine a LD50 and a NOEL for sublethal effects according to OPPTS Method 850.2100, in compliance with GLP. Ten quail, 5 males and 5 females, were assigned to each of the treatment groups and the control group by indiscriminate draw. The test consisted of a geometric series of five dosage groups and a control group. Birds were acclimated to the study room and caging for ten weeks prior to test initiation. The birds were fasted for approximately 18 h prior to dosing. At experimental start, a single dose of 0, 175, 292, 486, 810 or 1350 mg/kg bw of the test substance in diluent was orally intubated directly into the crop or proventriculus of each bird. From test initiation until termination, all birds were observed at least twice daily. A record was maintained of all mortality, signs of toxicity and abnormal behavior. Body weights were measured individually at the initiation of the test and on Days 3, 7, and 14. Average feed consumption was determined by pen for each dosage group and the control group for Days 0-3, 4-7 and 8-14. Under the study conditions, the acute oral LD50 value for northern bobwhite exposed to the test substance as a single oral dose was calculated to be 561 mg/kg bw, with a 95% confidence interval of 441 to 716 mg/kg bw (Watson, 2010).