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EC number: 208-021-9 | CAS number: 505-84-0
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Repeated dose toxicity: inhalation
Administrative data
- Endpoint:
- sub-chronic toxicity: inhalation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 04 APRIL 2019 to 11 SEPTEMBER 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 021
- Report date:
- 2021
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 413 (90-Day (Subchronic) Inhalation Toxicity Study
- Version / remarks:
- 25 June 2018
- Deviations:
- yes
- Remarks:
- Some observed chamber temperatures and humidity were below the recommended range (recorded values 14.5 - 16.6 °C). These deviations had no impact on study integrity.
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- Diethoxymethane
- EC Number:
- 207-330-6
- EC Name:
- Diethoxymethane
- Cas Number:
- 462-95-3
- Molecular formula:
- C5H12O2
- IUPAC Name:
- Diethoxymethane
- Test material form:
- liquid
- Remarks:
- Clear liquid
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Lambiotte & Cie - 1808131400R and 1904111400R
- Expiration date of the lot/batch: 13 August 2020 and 11 April 2021
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Refrigerated (2 to 8°C), under nitrogen and protected from
light
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on species / strain selection:
- The RccHan™®:WIST strain was used because of the historical control data available at this laboratory.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Envigo RMS Limited (UK).
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 7 to 8 weeks.
- Weight at study initiation: Males: 193 to 240 g; Females: 143 to 178 g
- Housing: Five of the same sex, unless reduced by mortality or isolation
- Diet: Teklad 2014C Diet ad libitum (removed overnight before blood sampling for hematology or blood chemistry)
- Water: Potable water from the public supply via polycarbonate bottles with sipper tubes, ad libitum (except during exposure)
- Acclimation period: At least 11 days before commencement of treatment.
DETAILS OF FOOD AND WATER QUALITY: Certificates of analysis for the diet were scrutinized and approved before any batch of diet was released for use. Certificates of analysis are routinely provided by the water supplier.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Monitored and maintained within the range of 20-24ºC
- Humidity (%): Monitored and maintained within the range of 40-70%.
- Air changes (per hr): Filtered fresh air which was passed to atmosphere and not recirculated.
- Photoperiod (hrs dark / hrs light): Artificial lighting, 12 hours dark : 12 hours light.
IN-LIFE DATES: From: 04 APRIL 2019 To: 15 to 17 JULY 2019
Administration / exposure
- Route of administration:
- inhalation: aerosol
- Type of inhalation exposure:
- snout only
- Vehicle:
- clean air
- Remarks on MMAD:
- Due to the physical nature of the test material atmosphere it was not possible to capture the samples required to determine a particle size distribution
- Details on inhalation exposure:
- GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Flow through nose-only chamber; Aluminum alloy construction comprising a base unit, three animal exposure sections, a top section and a pre-chamber
- Method of holding animals in test chamber: Plastic nose-only restraint tube
- Source and rate of air: From in-house compressed air system – breathing quality
- System of generating particulates/aerosols: Glass sintered vaporizer; The test item was supplied to the generator, via a feed line, from a syringe driven at a constant rate by a syringe pump
- Temperature, humidity in air chamber: 19 – 25°C; 30 - 70%
- Air flow rate: Generator flow: 19L/minute; Extract flow: 20L/minute
- Treatment of exhaust air: Drawn by in-house vacuum system and filtered locally - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Test sample (Solvent trap collected in trapping solvent): Transfer to volumetric flask using extraction solvent. Further dilute, if necessary, using extraction solvent to provide a solution containing Ethylal at a nominal concentration in the range 20 – 200 µg/mL. Inject onto the GC in duplicate.
- Duration of treatment / exposure:
- 13 weeks 6 hours daily
- Frequency of treatment:
- exposure for 5 days per week
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 ppm (analytical)
- Dose / conc.:
- 505 ppm (analytical)
- Dose / conc.:
- 1 520 ppm (analytical)
- Dose / conc.:
- 3 860 ppm (analytical)
- No. of animals per sex per dose:
- 10
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: In a previous inhalation toxicity study, three groups of rats were exposed to ethylal for 2 weeks (6 hours/day, 5 days/week) at mean achieved aerosol concentrations of 490, 1520 and 3930 ppm respectively. A fourth group received a single 4-hour exposure at an achieved concentration of 4810 ppm.
Exposures at mean achieved concentrations of 490, 1520 or 3930 ppm resulted in clinical signs of unsteady gait/swaying, salivation and piloerection. These signs were transient after dosing and were not considered to be adverse for these groups. Pathological examination did not show any test item related changes at 490 or 1520 ppm. Minimal olfactory epithelial degeneration was evident in two males and one female exposed at 3930 ppm.
A single exposure to ethylal at an achieved concentration 4810 ppm was stopped after 4 hours, due to adverse clinical signs. Signs seen on return to home cage included: unresponsiveness to external stimuli, decreased activity, breathing abnormalities, reduced body temperature and abnormal posture. The condition of these animals did not improve by 90 minutes post exposure and they were euthanized on grounds of animal welfare. Pathological examination showed moderate olfactory epithelial degeneration in all animals.
For this study, the highest target concentration of 4000 ppm was expected to result in treatment related effects, but not cause deaths. The lower target concentrations of 500 and 1500 ppm were chosen to explore the dose relationship of any effects seen over 13 weeks. - Positive control:
- No
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: A detailed weekly physical examination was performed on each animal to monitor general health.
BODY WEIGHT: Yes
- Time schedule for examinations: The weight of each animal was recorded twice weekly one week before treatment commenced to Week 4, weekly from Weeks 5 to 13 and before necropsy.
FOOD CONSUMPTION AND COMPOUND INTAKE:
The weight of food supplied to each cage, that remaining and an estimate of any spilled was
recorded for the week before treatment started and for each week throughout the study.
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: The eyes of the animals were examined by means of a binocular indirect ophthalmoscope as follows: pretreatment for all animals and at week 13 for all animals of groups 1 and 4.
HAEMATOLOGY: Yes
- Time schedule for collection of blood: week 13
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all animals
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 13
- Animals fasted: Yes
- How many animals: all animals.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No
IMMUNOLOGY: No
BRONCHOALVEOLAR LAVAGE FLUID (BALF): Yes
- Time schedule for analysis: at termination
- Dose groups that were examined: all groups
- Number of animals: all animals
LUNG BURDEN: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes - Statistics:
- All statistical analyses were carried out separately for males and females using the individual
animal as the basic experimental unit.
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Unsteady gait was occasionally observed for rats receiving ethylal at all exposure levels. In addition, swaying, decreased activity, lacrimation and wet rales were occasionally observed for a proportion of rats exposed to 3860 ppm. These signs were observed on return to home cage and resolved by the end of the working day.
On Week.Days 7.1 to 7.3, clinical signs evident for rats receiving the highest exposure level of ethylal were more marked than those seen after other exposures. Signs seen after exposure, for a proportion of rats, included locomotor and responsiveness effects (decreased activity, unsteady gait, unresponsiveness, flattened posture) and breathing effects (slow and/or shallow breathing and wet rales). Animal 138 (a high exposure level female) was removed from exposure on Week.Day 7.2, after approximately 4.5 hours, due to welfare concerns regarding clinical signs. Clinical signs evident for this animal included abnormal gait (unsteady and elevated), hunched posture, shallow breathing, decreased activity and pale ears. This rat had recovered by the end of the working day.
Signs associated with the administration procedure included red staining of the head and eyes, and/or wet fur on occasion. These were seen for rats of all groups, including control, on return to the home cage and at the end of the working day. These are signs considered to be due to the method of restraint and are not test item related.
Table 9 includes individual observation following detailed weekly physical examination - Mortality:
- mortality observed, treatment-related
- Description (incidence):
- There was one unscheduled death due to overexposure to ethylal. Animal 134, a female receiving the highest exposure level of ethylal was found dead at the end of exposure on Week.Day 7.1. Macroscopic observations included incomplete deflation of the lungs, depressions and pale areas in the glandular mucosa of the stomach (not confirmed microscopically), and gaseous distension of the stomach, duodenum and jejunum. Incomplete deflation of the lungs and gaseous distension of the GIT were considered to be post-mortem changes. Other microscopic changes consisted in unilateral pyogranulomatous inflammation within the glandular epithelium of the nose (with presence of the bacteria and Splendore-Hoeppli material) and minimal bilateral dilation if the renal pelvis.
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Differences in mean bodyweight gain were small compared with intragroup variation and were inconsistent between exposure levels and/or sexes. Small weight losses in the final week, for all groups, are considered to be associated with blood sampling techniques.
Table 1 - Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- Minor fluctuations in food consumption lacked dose relationship, were inconsistent between the sexes and are considered to be unrelated to treatment.
Table 8 - Ophthalmological findings:
- no effects observed
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- All differences from control, that attained statistical significance, were small compared with intragroup variation, lacked dose relationship and/or were inconsistent between sexes.
Table 2 - Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- All differences from control, that attained statistical significance, were small compared with intragroup variation, lacked dose relationship and/or were inconsistent between sexes.
Table 3 - Organ weight findings including organ / body weight ratios:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Mean liver weights (absolute and adjusted for terminal bodyweight) were higher than control for males exposed to 1520 ppm and both sexes exposed to 3860 ppm (up to 1.23X control).
Mean kidney weights (absolute and adjusted for terminal bodyweight) were higher than control for females exposed to 1520 ppm and both sexes exposed to 3860 ppm (up to 1.21X control).
Mean adrenal weights (absolute and adjusted for terminal bodyweight) were higher than control for both sexes exposed to 1520 or 3860 ppm (up to 1.26X control).
All other differences from control were small, lacked dose relationship and/or were inconsistent between sexes.
Table 4 - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- Table 6
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Changes related to exposure to ethylal were seen in thyroid glands and liver.
In the thyroids, minimal hypertrophy of the follicular cells was observed for one control male and 5 out of 10 males exposed to 3860 ppm.
In the liver, diffuse minimal centrilobular hepatocellular hypertrophy was recorded for four males out of 10 exposed to 3860 ppm.
Table 7 - Other effects:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test item-related effects on BAL parameters.
All differences from control were small, lacked dose relationship and/or were inconsistent between sexes.
Table 5
Effect levels
- Key result
- Dose descriptor:
- NOAEC
- Effect level:
- 3 860 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- clinical biochemistry
- clinical signs
- food consumption and compound intake
- gross pathology
- haematology
- histopathology: non-neoplastic
- mortality
- ophthalmological examination
- organ weights and organ / body weight ratios
Target system / organ toxicity
- Critical effects observed:
- no
Any other information on results incl. tables
| Air control | Ethylal | ||
Dose group | 1 | 2 | 3 | 4 |
Exposure level (ppm) | 0 | 505 | 1520 | 3860 |
Table 1: Body weight – group mean values (g)
Group /Sex |
| Day P1 |
P4 | Day 1 |
4 |
8 |
11 |
15 |
18 |
22 |
25 |
29 |
36 |
43 |
Statistics test |
|
|
|
|
|
|
|
|
|
|
|
|
| |
1M | Mean | 187 | 203 | 217 | 219 | 234 | 238 | 253 | 256 | 271 | 272 | 285 | 299 | 310 |
| SD | 5.0 | 6.2 | 7.5 | 11.1 | 13.1 | 16.2 | 17.2 | 20.7 | 21.9 | 24.0 | 24.1 | 26.2 | 27.0 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 186 | 200 | 213 | 211 | 227 | 230 | 243 | 248 | 262 | 264 | 277 | 286 | 298 |
| SD | 5.8 | 6.5 | 7.8 | 8.6 | 10.8 | 11.7 | 13.9 | 13.8 | 16.4 | 17.3 | 17.7 | 19.7 | 21.0 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1M |
|
|
|
|
|
|
|
|
|
|
|
|
|
3M | Mean | 182 | 200 | 214 | 217 | 234 | 240 | 253 | 259 | 274 | 275 | 286 | 296 | 308 |
| SD | 4.9 | 7.0 | 9.5 | 10.8 | 12.6 | 13.3 | 15.3 | 15.4 | 17.6 | 19.9 | 21.1 | 21.4 | 22.8 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1M |
|
|
|
|
|
|
|
|
|
|
|
|
|
4M | Mean | 185 | 203 | 220 | 220 | 237 | 238 | 253 | 254 | 270 | 266 | 280 | 290 | 300 |
| SD | 7.6 | 9.8 | 11.0 | 10.9 | 13.6 | 14.6 | 18.0 | 18.7 | 21.0 | 20.8 | 22.0 | 23.8 | 25.0 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1M |
|
|
|
|
|
|
|
|
|
|
|
|
|
Table 1 (cont): Body weight – group mean values (g)
Group /Sex |
| Day 50 |
57 |
64 |
71 |
78 |
85 |
92 | Change 1-92 |
Statistics test |
|
|
|
|
|
|
| Wi | |
1M | Mean | 318 | 326 | 330 | 337 | 343 | 350 | 342 | 124 |
| SD | 29.3 | 31.7 | 33.2 | 36.0 | 36.4 | 37.9 | 40.3 | 34.5 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 301 | 311 | 320 | 324 | 328 | 335 | 330 | 117 |
| SD | 20.5 | 21.3 | 21.4 | 22.3 | 23.1 | 20.9 | 21.7 | 18.3 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1M |
|
|
|
|
|
|
| 94 |
3M | Mean | 311 | 323 | 330 | 335 | 340 | 348 | 344 | 129 |
| SD | 22.3 | 23.6 | 23.4 | 25.0 | 26.4 | 27.1 | 26.2 | 20.2 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1M |
|
|
|
|
|
|
| 104 |
4M | Mean | 302 | 311 | 318 | 324 | 331 | 337 | 333 | 113 |
| SD | 25.1 | 26.8 | 29.7 | 32.2 | 32.1 | 34.2 | 34.3 | 27.3 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1M |
|
|
|
|
|
|
| 91 |
Table 1 (cont): Body weight – group mean values (g)
Group /Sex |
| Day P1 |
P4 | Day 1 |
4 |
8 |
11 |
15 |
18 |
22 |
25 |
29 |
36 |
43 |
Statistics test |
|
|
|
|
|
|
|
|
|
|
|
|
| |
1F | Mean | 138 | 148 | 156 | 158 | 167 | 167 | 174 | 176 | 183 | 183 | 189 | 196 | 202 |
| SD | 6.2 | 8.4 | 9.1 | 8.4 | 8.4 | 11.0 | 12.1 | 12.2 | 12.2 | 14.6 | 13.6 | 12.6 | 15.7 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 140 | 149 | 156 | 160 | 167 | 165 | 174 | 174 | 180 | 182 | 187 | 194 | 197 |
| SD | 5.3 | 6.8 | 8.3 | 7.8 | 8.0 | 8.6 | 9.4 | 9.2 | 9.8 | 11.2 | 10.5 | 10.8 | 13.5 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1F |
|
|
|
|
|
|
|
|
|
|
|
|
|
3F | Mean | 144 | 155 | 164 | 166 | 175 | 175 | 182 | 185 | 193 | 193 | 200 | 206 | 211 |
| SD | 5.1 | 5.8 | 7.7 | 6.6 | 7.6 | 8.2 | 9.2 | 11.1 | 11.6 | 11.6 | 12.5 | 11.6 | 9.5 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1F |
|
|
|
|
|
|
|
|
|
|
|
|
|
4F | Mean | 140 | 148 | 157 | 159 | 167 | 164 | 169 | 172 | 179 | 178 | 180 | 189 | 195 |
| SD | 7.2 | 6.8 | 7.9 | 7.9 | 8.4 | 8.4 | 7.9 | 8.7 | 9.8 | 10.4 | 9.0 | 11.5 | 8.1 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1F |
|
|
|
|
|
|
|
|
|
|
|
|
|
Table 1 (cont): Body weight – group mean values (g)
Group /Sex |
| Day 50 |
57 |
64 |
71 |
78 |
85 |
92 | Change 1-92 |
Statistics test |
|
|
|
|
|
|
| Wi | |
1F | Mean | 203 | 206 | 210 | 212 | 211 | 215 | 213 | 58 |
| SD | 16.7 | 17.4 | 15.5 | 17.8 | 18.3 | 19.2 | 17.1 | 13.2 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 199 | 205 | 207 | 206 | 208 | 211 | 209 | 52 |
| SD | 12.3 | 13.8 | 13.5 | 14.2 | 14.6 | 13.0 | 15.5 | 8.6 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1F |
|
|
|
|
|
|
| 90 |
3F | Mean | 213 | 217 | 220 | 220 | 221 | 225 | 221 | 57 |
| SD | 12.5 | 13.9 | 12.5 | 10.9 | 13.9 | 14.0 | 9.7 | 7.9 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| % of 1F |
|
|
|
|
|
|
| 99 |
4F | Mean | 197 | 201 | 201 | 204 | 206 | 209 | 206 | 49 |
| SD | 13.4 | 8.8 | 13.2 | 11.2 | 12.5 | 9.6 | 12.1 | 8.9 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
| % of 1F |
|
|
|
|
|
|
| 85 |
Table 2: Hematology – group mean values during Week 13 of treatment
Group /Sex |
| Hct L/L | Hb g/dL | RBC x 1012/L | Retic x 1012/L | MCH pg | MCHC g/dL | MCV fL | RDW % |
Statistics test | Wi | Wi | Wi | Wi | Wi | Wi | Wi | Wi | |
1M | Mean | 0.499 | 15.7 | 8.90 | 0.118 | 17.7 | 31.5 | 56.1 | 11.3 |
| SD | 0.0121 | 0.45 | 0.304 | 0.0212 | 0.56 | 0.55 | 1.42 | 0.47 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 0.497 | 15.7 | 8.94 | 0.115 | 17.5 | 31.5 | 55.7 | 11.6 |
| SD | 0.0206 | 0.63 | 0.438 | 0.0141 | 0.70 | 0.70 | 1.46 | 0.44 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3M | Mean | 0.494 | 15.6 | 8.79 | 0.129 | 17.7 | 31.5 | 56.2 | 11.6 |
| SD | 0.0103 | 0.37 | 0.293 | 0.0193 | 0.64 | 0.54 | 1.83 | 0.58 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4M | Mean | 0.490 | 15.5 | 8.71 | 0.134 | 17.9 | 31.7 | 56.3 | 11.9* |
| SD | 0.0134 | 0.41 | 0.358 | 0.0230 | 0.54 | 0.29 | 1.50 | 0.45 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Table 2 (cont): Hematology – group mean values during Week 13 of treatment
Group /Sex |
| WBC x 109/L | N x 109/L | L x 109/L | E x 109/L | B x 109/L | M x 109/L | LUC x 109/L |
Statistics test | Wi | Wi | lWi | Wi | Wi | Wi | Wi | |
1M | Mean | 4.95 | 0.94 | 3.83 | 0.05 | 0.01 | 0.09 | 0.03 |
| SD | 1.320 | 0.474 | 0.859 | 0.025 | 0.006 | 0.045 | 0.012 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 5.21 | 1.15 | 3.87 | 0.05 | 0.01 | 0.11 | 0.02 |
| SD | 1.035 | 0.207 | 0.920 | 0.018 | 0.007 | 0.048 | 0.016 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3M | Mean | 5.84 | 1.28 | 4.32 | 0.07 | 0.01 | 0.12 | 0.03 |
| SD | 1.619 | 0.708 | 1.231 | 0.039 | 0.011 | 0.054 | 0.021 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4M | Mean | 5.34 | 0.98 | 4.17 | 0.06 | 0.01 | 0.11 | 0.03 |
| SD | 0.856 | 0.253 | 0.904 | 0.014 | 0.003 | 0.037 | 0.012 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Table 2 (cont): Hematology – group mean values during Week 13 of treatment
Group /Sex |
| Plt x 109/L | PT sec | APTT sec |
Statistics test |
|
|
| |
1M | Mean | 823 | 21.3 | 14.3 |
| SD | 125.7 | 0.74 | 0.67 |
| N | 10 | 10 | 10 |
2M | Mean | 857 | 20.7 | 14.8 |
| SD | 104.7 | 0.54 | 1.23 |
| N | 10 | 9 | 9 |
3M | Mean | 824 | 20.6 | 14.5 |
| SD | 133.9 | 1.28 | 1.03 |
| N | 10 | 10 | 10 |
4M | Mean | 824 | 20.3* | 14.4 |
| SD | 70.6 | 0.90 | 0.97 |
| N | 10 | 10 | 10 |
Table 2 (cont): Hematology – group mean values during Week 13 of treatment
Group /Sex |
| Hct L/L | Hb g/dL | RBC x 1012/L | Retic x 1012/L | MCH pg | MCHC g/dL | MCV fL | RDW % |
Statistics test | Wi | Wi | Wi | Wi | Wi | Sh | Wi | Wi | |
1F | Mean | 0.465 | 14.7 | 8.00 | 0.125 | 18.3 | 31.5 | 58.2 | 10.2 |
| SD | 0.0144 | 0.50 | 0.243 | 0.0185 | 0.74 | 1.03 | 0.97 | 0.40 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 0.465 | 14.7 | 8.00 | 0.125 | 18.3 | 31.5 | 58.2 | 10.2 |
| SD | 0.0144 | 0.50 | 0.243 | 0.0185 | 0.74 | 1.03 | 0.97 | 0.40 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3F | Mean | 0.465 | 14.7 | 8.04 | 0.122 | 18.3 | 31.6 | 57.9 | 10.2 |
| SD | 0.0107 | 0.36 | 0.225 | 0.0197 | 0.37 | 0.45 | 0.95 | 0.25 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4F | Mean | 0.459 | 14.6 | 7.96 | 0.118 | 18.3 | 31.8 | 57.7 | 10.7** |
| SD | 0.0148 | 0.46 | 0.433 | 0.0199 | 0.64 | 0.48 | 1.68 | 0.27 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
Table 2 (cont): Hematology – group mean values during Week 13 of treatment
Group /Sex |
| WBC x 109/L | N x 109/L | L x 109/L | E x 109/L | B x 109/L | M x 109/L | LUC x 109/L |
Statistics test | Wi | Wi | Wi | Wi | Wi | Wi | Wi | |
1F | Mean | 3.50 | 0.57 | 2.78 | 0.06 | 0.00 | 0.07 | 0.02 |
| SD | 0.927 | 0.183 | 0.857 | 0.033 | 0.005 | 0.024 | 0.007 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 4.48* | 0.60 | 3.74 | 0.05 | 0.01 | 0.07 | 0.02 |
| SD | 1.031 | 0.218 | 0.893 | 0.012 | 0.005 | 0.045 | 0.011 |
| N | 10 | 10# | 10# | 10# | 10# | 10# | 9 |
3F | Mean | 4.24* | 0.75 | 3.33 | 0.05 | 0.01 | 0.08 | 0.02 |
| SD | 0.742 | 0.248 | 0.650 | 0.019 | 0.009 | 0.027 | 0.008 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4F | Mean | 4.30* | 0.75 | 3.39 | 0.04 | 0.00 | 0.09 | 0.02 |
| SD | 0.614 | 0.192 | 0.479 | 0.018 | 0.005 | 0.029 | 0.010 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
# Manual counts included
Table 2 (cont): Hematology – group mean values during Week 13 of treatment
Group /Sex |
| Plt x 109/L | PT sec | APTT sec |
Statistics test | Wi | Sh | Wi | |
1F | Mean | 818 | 22.8 | 11.3 |
| SD | 106.2 | 3.41 | 2.01 |
| N | 10 | 10 | 10 |
2F | Mean | 891 | 21.8 | 11.2 |
| SD | 120.6 | 0.91 | 1.67 |
| N | 9 | 10 | 10 |
3F | Mean | 801 | 21.8 | 10.8 |
| SD | 76.3 | 1.31 | 0.95 |
| N | 10 | 10 | 10 |
4F | Mean | 861 | 20.7* | 10.8 |
| SD | 149.7 | 1.14 | 1.52 |
| N | 9 | 9 | 9 |
Table 3: Blood chemistry – group mean values during Week 13 of treatment
Group /Sex |
| ALP U/L | ALT U/L | AST U/L | Bili µmol/L | Urea mmol/L | BUN mmol/L | Creat µmol/L | Gluc mmol/L | Chol mmol/L | Trig mmol/L |
Statistics test | lWi | Wi | Wi | Du | Sh | Sh | Wi | Wi | Wi | Wi | |
1M | Mean | 74 | 48 | 83 | 2 | 5.89 | 5.89 | 28 | 6.59 | 2.37 | 0.85 |
| SD | 17.6 | 10.8 | 26.3 | 0.4 | 0.792 | 0.792 | 1.9 | 0.791 | 0.213 | 0.180 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 76 | 44 | 79 | 2 | 6.62 | 6.62 | 28 | 6.97 | 2.30 | 0.80 |
| SD | 7.3 | 6.6 | 14.7 | 0.4 | 0.673 | 0.673 | 1.3 | 1.169 | 0.316 | 0.376 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3M | Mean | 62 | 49 | 80 | 1* | 6.09 | 6.09 | 28 | 6.71 | 2.42 | 0.93 |
| SD | 7.1 | 12.5 | 14.3 | 0.5 | 0.653 | 0.653 | 2.8 | 1.021 | 0.318 | 0.385 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4M | Mean | 76 | 60* | 96 | 2 | 6.60 | 6.60 | 29 | 6.50 | 2.22 | 0.80 |
| SD | 18.3 | 13.2 | 37.3 | 0.4 | 1.919 | 1.919 | 2.2 | 1.032 | 0.182 | 0.292 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Table 3 (cont): Blood chemistry – group mean values during Week 13 of treatment
Group /Sex |
| Na mmol/L | K mmol/L | Cl mmol/L | Ca mmol/L | Phos mmol/L | Total Prot g/L | Alb g/L | A/G Ratio |
Statistics test | Wi | Wi | Wi | Wi | Wi | Wi | Wi | Wi | |
1M | Mean | 144 | 3.86 | 103.6 | 2.64 | 1.83 | 68 | 39 | 1.34 |
| SD | 1.5 | 0.163 | 1.51 | 0.080 | 0.166 | 2.5 | 0.7 | 0.114 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 143 | 4.00 | 101.9** | 2.66 | 1.98 | 69 | 39 | 1.32 |
| SD | 1.4 | 0.114 | 1.52 | 0.056 | 0.146 | 2.2 | 1.1 | 0.071 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3M | Mean | 143 | 4.02* | 101.5** | 2.67 | 1.93 | 70 | 39 | 1.27 |
| SD | 0.7 | 0.199 | 0.68 | 0.062 | 0.079 | 1.4 | 0.9 | 0.077 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4M | Mean | 144 | 4.24** | 102.5** | 2.72* | 2.06** | 68 | 39 | 1.34 |
| SD | 1.1 | 0.164 | 0.99 | 0.082 | 0.155 | 2.1 | 1.3 | 0.078 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Table 3 (cont): Blood chemistry – group mean values during Week 13 of treatment
Group /Sex |
| ALP U/L | ALT U/L | AST U/L | Bili µmol/L | Urea mmol/L | BUN mmol/L | Creat µmol/L | Gluc mmol/L | Chol mmol/L | Trig mmol/L |
Statistics test | Wi | Wi | Wi | Fe | Wi | Wi | Wi | Wi | Wi | Wi | |
1F | Mean | 40 | 45 | 81 | 2 | 7.77 | 7.77 | 31 | 6.04 | 2.16 | 0.76 |
| SD | 12.3 | 10.1 | 16.6 | 0.4 | 0.985 | 0.985 | 3.8 | 0.914 | 0.383 | 0.371 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 35 | 51 | 82 | 2 | 8.39 | 8.39 | 32 | 5.66 | 2.29 | 0.91 |
| SD | 10.2 | 10.9 | 20.7 | 0.3 | 1.064 | 1.064 | 2.6 | 0.410 | 0.421 | 0.253 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3F | Mean | 34 | 48 | 75 | 2 | 7.45 | 7.45 | 27* | 6.50 | 2.38 | 1.02 |
| SD | 9.2 | 9.1 | 15.2 | 0.6 | 1.131 | 1.131 | 2.9 | 1.193 | 0.558 | 0.407 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4F | Mean | 40 | 68** | 75 | 2 | 7.32 | 7.32 | 27** | 6.87 | 2.47 | 0.76 |
| SD | 10.2 | 20.1 | 11.4 | 0.4 | 0.962 | 0.962 | 3.2 | 1.029 | 0.312 | 0.194 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
Table 3 (cont): Blood chemistry – group mean values during Week 13 of treatment
Group /Sex |
| Na mmol/L | K mmol/L | Cl mmol/L | Ca mmol/L | Phos mmol/L | Total Prot g/L | Alb g/L | A/G Ratio |
Statistics test | Wi | Wi | Wi | Wi | Wi | Wi | Wi | Wi | |
1F | Mean | 141 | 3.63 | 102.0 | 2.63 | 1.67 | 70 | 42 | 1.49 |
| SD | 1.0 | 0.240 | 1.26 | 0.067 | 0.189 | 4.0 | 2.0 | 0.103 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 142 | 3.73 | 101.9 | 2.73** | 1.80 | 73 | 43 | 1.42 |
| SD | 1.1 | 0.139 | 1.11 | 0.050 | 0.185 | 2.0 | 1.4 | 0.064 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3F | Mean | 143 | 3.64 | 101.7 | 2.74** | 1.77 | 73 | 43 | 1.40 |
| SD | 0.8 | 0.157 | 1.36 | 0.088 | 0.200 | 3.0 | 1.9 | 0.100 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4F | Mean | 142 | 3.77 | 101.2 | 2.68** | 1.98** | 69 | 40* | 1.40 |
| SD | 1.2 | 0.175 | 1.65 | 0.052 | 0.129 | 2.7 | 1.2 | 0.088 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
Table 4: Organ weights – group mean absolute and adjusted values (g) for animals killed after 13 weeks of treatment
Group /Sex |
|
Terminal Body weight |
Adrenals |
Brain |
Epididymides |
Heart |
Kidneys |
Liver |
Lungs and Bronchi |
Spleen |
Testes |
Statistics test | Wi |
|
|
|
|
|
|
|
|
| |
1M | Mean | 341.3 | 0.061 | 1.955 | 0.984 | 0.977 | 1.768 | 10.930 | 1.310 | 0.667 | 2.704 |
| SD | 40.5 | 0.008 | 0.108 | 0.159 | 0.114 | 0.292 | 1.850 | 0.204 | 0.126 | 0.319 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 331.1 | 0.063 | 2.017 | 1.023 | 0.981 | 1.746 | 11.070 | 1.294 | 0.647 | 2.696 |
| SD | 21.8 | 0.011 | 0.104 | 0.153 | 0.128 | 0.173 | 1.252 | 0.156 | 0.096 | 0.559 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3M | Mean | 343.6 | 0.069 | 2.015 | 1.013 | 1.047 | 1.934 | 13.154 | 1.317 | 0.706 | 2.921 |
| SD | 25.5 | 0.008 | 0.125 | 0.125 | 0.155 | 0.248 | 1.527 | 0.088 | 0.127 | 0.476 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4M | Mean | 332.3 | 0.074 | 2.093 | 0.942 | 1.010 | 2.137 | 13.102 | 1.372 | 0.641 | 2.709 |
| SD | 34.0 | 0.009 | 0.115 | 0.140 | 0.192 | 0.728 | 1.773 | 0.077 | 0.084 | 0.457 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 9 | 10 | 10 |
Statistics test | Wi | Wi | Wi | Wi | lWi | Wi | Wi | Wi | Wi | ||
1M | Adjusted Mean | 0.061 | 1.948 | 0.976 | 0.969 | 1.727 | 10.767 | 1.298 | 0.661 | 2.680 | |
2M | Adjusted Mean | 0.063 | 2.026 | 1.034 | 0.992 | 1.770 | 11.300 | 1.311 | 0.656 | 2.730 | |
3M | Adjusted Mean | 0.069 | 2.005 | 1.001 | 1.035 | 1.875 | 12.903** | 1.298 | 0.697 | 2.884 | |
4M | Adjusted Mean | 0.074** | 2.101** | 0.951 | 1.018 | 2.092** | 13.285** | 1.388 | 0.648 | 2.736 |
Table 4 (cont): Organ weights – group mean absolute and adjusted values (g) for animals killed after 13 weeks of treatment
Group /Sex |
| Terminal Body weight | Thymus and Thyroids | Parathyroids |
Statistics test | Wi |
|
| |
1M | Mean | 341.3 | 0.237 | 0.022 |
| SD | 40.5 | 0.087 | 0.007 |
| N | 10 | 10 | 10 |
2M | Mean | 331.1 | 0.192 | 0.020 |
| SD | 21.8 | 0.041 | 0.004 |
| N | 10 | 10 | 10 |
3M | Mean | 343.6 | 0.226 | 0.019 |
| SD | 25.5 | 0.048 | 0.004 |
| N | 10 | 10 | 10 |
4M | Mean | 332.3 | 0.198 | 0.021 |
| SD | 34.0 | 0.057 | 0.004 |
| N | 10 | 10 | 10 |
Statistics test | Wi | Wi | ||
1M | Adjusted Mean | 0.234 | 0.022 | |
2M | Adjusted Mean | 0.197 | 0.020 | |
3M | Adjusted Mean | 0.221 | 0.019 | |
4M | Adjusted Mean | 0.201 | 0.021 |
Table 4 (cont): Organ weights – group mean absolute and adjusted values (g) for animals killed after 13 weeks of treatment
Group /Sex |
|
Terminal Body weight |
Adrenals |
Brain |
Heart |
Kidneys |
Liver |
Lungs and Bronchi |
Ovaries | Spleen | Thymus |
Statistics test | Wi |
|
|
|
|
| St |
|
|
| |
1F | Mean | 212.5 | 0.066 | 1.928 | 0.740 | 1.252 | 7.656 | 1.012 | 0.079 | 0.512 | 0.199 |
| SD | 19.2 | 0.011 | 0.115 | 0.135 | 0.097 | 0.780 | 0.126 | 0.016 | 0.096 | 0.063 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 209.1 | 0.071 | 1.923 | 0.692 | 1.274 | 7.572 | 1.025 | 0.083 | 0.527 | 0.229 |
| SD | 14.5 | 0.006 | 0.072 | 0.104 | 0.091 | 0.560 | 0.066 | 0.016 | 0.094 | 0.041 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3F | Mean | 221.1 | 0.081 | 1.942 | 0.791 | 1.386 | 8.316 | 1.108 | 0.084 | 0.520 | 0.224 |
| SD | 10.8 | 0.007 | 0.108 | 0.103 | 0.113 | 0.700 | 0.088 | 0.014 | 0.056 | 0.059 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4F | Mean | 206.5 | 0.083 | 1.870 | 0.675 | 1.369 | 8.710 | 0.996 | 0.088 | 0.493 | 0.180 |
| SD | 11.5 | 0.008 | 0.118 | 0.045 | 0.099 | 0.787 | 0.020 | 0.021 | 0.069 | 0.046 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
Statistics test | Wi | Wi | Wi | Wi | Wi |
| Wi | Wi | Wi | ||
1F | Adjusted Mean | 0.066 | 1.928 | 0.740 | 1.251 | 7.653 |
| 0.079 | 0.512 | 0.199 | |
2F | Adjusted Mean | 0.071 | 1.932 | 0.706 | 1.286 | 7.686 |
| 0.085 | 0.538 | 0.233 | |
3F | Adjusted Mean | 0.081** | 1.919 | 0.755 | 1.355* | 8.025 |
| 0.079 | 0.491 | 0.212 | |
4F | Adjusted Mean | 0.083** | 1.883 | 0.700 | 1.391** | 8.910** |
| 0.091 | 0.512 | 0.189 |
Table 4 (cont): Organ weights – group mean absolute and adjusted values (g) for animals killed after 13 weeks of treatment
Group /Sex |
| Terminal Body weight | Thymus and Thyroids | Parathyroids |
Statistics test | Wi |
|
| |
1F | Mean | 212.5 | 0.018 | 0.732 |
| SD | 19.2 | 0.006 | 0.234 |
| N | 10 | 10 | 10 |
2F | Mean | 209.1 | 0.017 | 0.676 |
| SD | 14.5 | 0.006 | 0.178 |
| N | 10 | 10 | 10 |
3F | Mean | 221.1 | 0.017 | 0.638 |
| SD | 10.8 | 0.003 | 0.171 |
| N | 10 | 10 | 10 |
4F | Mean | 206.5 | 0.016 | 0.598 |
| SD | 11.5 | 0.003 | 0.231 |
| N | 9 | 9 | 9 |
Statistics test | Wi | Wi | ||
1F | Adjusted Mean | 0.018 | 0.732 | |
2F | Adjusted Mean | 0.018 | 0.680 | |
3F | Adjusted Mean | 0.015 | 0.626 | |
4F | Adjusted Mean | 0.017 | 0.606 |
Table 5: Bronchoalveolar lavage – group mean differential cell counts in the right lung
Group /Sex |
| Different cell counts calculated per animal (106/animal) | Different cell counts as a percentage of total white blood cells (%) | |||||||
Total Cells | Neutrophils | Lymphocytes | Mononuclear cells | Eosinophils | Neutrophils | Lymphocytes | Mononuclear cells | Eosinophils | ||
Statistics test | lDu | sWi | sDu | Du | Wi | Wi | Wi | Wi | Wi | |
1M | Mean | 3.60 | 0.57 | 0.26 | 2.70 | 0.07 | 14.45 | 6.77 | 76.00 | 1.92 |
| SD | 1.64 | 0.44 | 0.21 | 1.16 | 0.05 | 8.15 | 3.34 | 10.34 | 1.22 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2M | Mean | 1.40*** | 0.13* | 0.06** | 1.20*** | 0.02 | 8.19 | 4.64 | 85.46 | 1.71 |
| SD | 0.48 | 0.12 | 0.05 | 0.45 | 0.03 | 7.23 | 3.52 | 9.18 | 2.57 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3M | Mean | 1.57** | 0.17* | 0.13 | 1.23*** | 0.04 | 9.06 | 7.31 | 81.07 | 2.56 |
| SD | 0.84 | 0.24 | 0.18 | 0.64 | 0.02 | 10.10 | 6.82 | 14.63 | 2.37 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
4M | Mean | 2.89 | 0.40* | 0.21 | 2.23 | 0.05 | 10.89 | 7.10 | 79.56 | 1.49 |
| SD | 1.46 | 0.56 | 0.12 | 0.90 | 0.04 | 8.37 | 2.35 | 8.99 | 1.21 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Table 5 (cont): Bronchoalveolar lavage – group mean differential cell counts in the right lung
Group /Sex |
| Different cell counts calculated per animal (106/animal) | Different cell counts as a percentage of total white blood cells (%) | |||||||
Total Cells | Neutrophils | Lymphocytes | Mononuclear cells | Eosinophils | Neutrophils | Lymphocytes | Mononuclear cells | Eosinophils | ||
Statistics test | Wi | Wi | sWi | Wi | Wi | Wi | Wi | Wi | Wi | |
1F | Mean | 1.85 | 0.26 | 0.13 | 1.43 | 0.03 | 11.84 | 4.88 | 81.57 | 1.33 |
| SD | 1.08 | 0.27 | 0.20 | 0.70 | 0.03 | 6.41 | 5.42 | 10.53 | 1.75 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
2F | Mean | 1.99 | 0.35 | 0.14 | 1.48 | 0.03 | 13.09 | 6.71 | 78.97 | 0.69 |
| SD | 1.36 | 0.43 | 0.11 | 0.83 | 0.04 | 9.92 | 2.92 | 12.68 | 1.14 |
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
3F | Mean | 1.89 | 0.16 | 0.12 | 1.59 | 0.02 | 7.12 | 5.62 | 85.93 | 0.90 |
| SD | 0.78 | 0.16 | 0.08 | 0.56 | 0.04 | 6.89 | 3.02 | 8.87 | 1.43 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
4F | Mean | 2.18 | 0.33 | 0.14 | 1.67 | 0.05 | 13.51 | 6.81 | 77.14 | 1.80 |
| SD | 1.02 | 0.25 | 0.06 | 0.74 | 0.05 | 8.66 | 2.91 | 11.87 | 1.79 |
| N | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 | 9 |
Table 5 (cont): Bronchoalveolar lavage – group mean values at termination
Group /Sex |
| LDH U/L | Prot g/L |
Statistics test | Wi | lWi | |
1M | Mean | 457 | 0.65 |
| SD | 341.3 | 0.659 |
| N | 10 | 10 |
2M | Mean | 228 | 0.17 |
| SD | 132.1 | 0.084 |
| N | 10 | 10 |
3M | Mean | 263 | 0.30 |
| SD | 184.2 | 0.466 |
| N | 10 | 10 |
4M | Mean | 297 | 0.25 |
| SD | 164.5 | 0.146 |
| N | 10 | 10 |
Table 5 (cont): Bronchoalveolar lavage – group mean values at termination
Group /Sex |
| LDH U/L | Prot g/L |
Statistics test | Wi | Wi | |
1F | Mean | 274 | 0.23 |
| SD | 147.7 | 0.188 |
| N | 10 | 10 |
2F | Mean | 367 | 0.50 |
| SD | 248.0 | 0.603 |
| N | 10 | 10 |
3F | Mean | 186 | 0.27 |
| SD | 150.8 | 0.445 |
| N | 10 | 10 |
4F | Mean | 385 | 0.47 |
| SD | 276.6 | 0.446 |
| N | 9 | 9 |
Table 6: Macropathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Number of animals within normal limits |
| 9 | 7 | 6 | 8 | 9 | 8 | 6 | 7 |
Epididymides Small |
1 |
0 |
0 |
0 |
- |
- |
- |
- | |
Kidneys Cyst(s) Dilated Pelvis Enlarged |
0 0 0 |
0 0 0 |
0 0 0 |
0 1 1 |
0 1 0 |
0 0 0 |
1 0 0 |
1 0 0 | |
Lungs and Bronchi Pale area(s) |
0 |
1 |
1 |
0 |
1 |
2 |
3 |
1 | |
Lymph Node, Mediastinal Abnormal color Enlarged |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
1 1 | |
Prostate Abnormal color |
0 |
1 |
0 |
0 |
- |
- |
- |
- | |
Skin and subcutis Scab(s) |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
Table 6 (cont): Macropathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Testes Abnormal color Pale area(s) Small Soft |
|
0 0 1 1 |
1 1 1 1 |
1 0 2 3 |
0 0 1 0 |
- - - - |
- - - - |
- - - - |
- - - - |
Ureters Distended |
|
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
Table 7: Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Adrenals | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Aorta Thoracic | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Bone, Femur incl Marrow | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Bone, Sternum Including Marrow | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Brain | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Brain, Olfactory Bulbs | No. examined | 10 | 0 | 0 | 7 | 10 | 0 | 0 | 9 |
Cecum | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Colon | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Duodenum | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Epididymides Sperm, Reduced, Luminal
Edema | No. examined Minimal Slight Moderate Total
Slight Total | 10 4 0 1 5
0 0 | 0 0 0 0 0
0 0 | 0 0 0 0 0
0 0 | 10 3 2 0 5
1 1 | - - - - -
- - | - - - - -
- - | - - - - -
- - | - - - - -
- - |
Epiglottis | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Esophagus | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Eyes Rosettes/Folds, Retina
| No. examined Minimal Total | 10 0 0 | 0 0 0 | 0 0 0 | 10 0 0 | 10 0 0 | 0 0 0 | 0 0 0 | 9 1 1 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Harderian Glands | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Heart Cardiomyopathy
Infiltrate, Inflammatory Cell | No. examined Minimal Total
Minimal Total | 10 1 1
1 1 | 0 0 0
0 0 | 0 0 0
0 0 | 10 0 0
0 0 | 10 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 9 0 0
0 0 |
Ileum | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Jejunum | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Kidneys Dilatation, Pelvic | No. examined Minimal Slight Moderate Marked Total | 10 1 0 0 0 1 | 0 0 0 0 0 0 | 0 0 0 0 0 0 | 10 1 1 0 1 3 | 10 0 0 1 0 1 | 0 0 0 0 0 0 | 0 0 0 0 0 0 | 9 0 0 0 0 0 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Lacrimal Glands | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Larynx Granuloma | No. examined Slight Total | 10 1 1 | 10 0 0 | 10 0 0 | 10 0 0 | 10 0 0 | 10 0 0 | 10 0 0 | 9 0 0 |
Liver Pigment, Hepatocellular
Hypertrophy, Hepatocellular, Centrilobular | No. examined Slight Total
Minimal Total | 10 1 1
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 10 1 1
5 5 | 10 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 9 0 0
0 0 |
Lungs and Bronchi Alveolar Macrophage Aggregation
Inflammation | No. examined Minimal Total
Minimal Total | 10 0 0
0 0 | 10 1 1
0 0 | 10 2 2
1 1 | 10 1 1
0 0 | 10 2 2
0 0 | 10 2 2
0 0 | 10 0 0
0 0 | 9 1 1
0 0 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Lymph Node, Axillary Lt Plasmocytosis | No. examined Minimal Slight Total | 10 0 0 0 | 0 0 0 0 | 0 0 0 0 | 10 0 0 0 | 10 3 0 3 | 0 0 0 0 | 0 0 0 0 | 9 2 1 3 |
Lymph Node, Mediastinal Mastocytosis
Plasmacytosis | No. examined Slight Total
Minimal Total | 0 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 0 0 0
0 0 | 1 1 1
1 1 |
Lymph Node, Tracheobronchial Erythrocytes, Intrasinusoidal
Plasmacytosis | No. examined Minimal Slight Total
Minimal Total | 10 0 0 0
0 0 | 10 0 0 0
0 0 | 10 0 0 0
0 0 | 10 0 0 0
0 0 | 10 0 0 0
0 0 | 9 0 1 1
0 0 | 10 0 0 0
2 2 | 9 1 0 1
0 0 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Mammary | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Nerve, Optic | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Nerve, Sciatic | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Nose/Turbinates | No. examined | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 9 |
Ovaries | No. examined | - | - | - | - | 10 | 0 | 0 | 9 |
Pancreas | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Parathyroids | No. examined | 7 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Pharynx, Nasal | No. examined | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 9 |
Pituitary | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Prostate Infiltrate, Inflammatory Cell
Inflammation | No. examined Minimal Moderate Marked Total
Marked Total | 10 1 0 1 2
0 0 | 1 0 1 0 1
1 1 | 0 0 0 0 0
0 0 | 10 0 0 0 0
0 0 | - - - - -
- - | - - - - -
- - | - - - - -
- - | - - - - -
- - |
Rectum | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Salivary Gland, Mandibular | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Salivary Gland, Sublingual | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Seminal Vesicles | No. examined | 10 | 0 | 0 | 10 | - | - | - | - |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Skeletal Muscle | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Skin and Subcutis Scab(s) | No. examined Minimal Total | 10 0 0 | 0 0 0 | 1 1 1 | 10 0 0 | 10 0 0 | 0 0 0 | 0 0 0 | 9 1 1 |
Spinal Cord Cervical | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Spinal Cord Lumbar | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Spinal Cord Thoracic | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Spleen | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Stomach | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Teeth | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Testes Degenration/Atrophy, Tubular | No. examined Minimal Slight Moderate Total | 10 5 1 1 7 | 1 0 1 0 1 | 4 1 0 3 4 | 10 3 6 0 9 | - - - - - | - - - - - | - - - - - | - - - - - |
Thymus | No. examined | 10 | 0 | 0 | 10 | 10 | 0 | 0 | 9 |
Thyroids Hypertrophy, Follicular Cells | No. examined Minimal Total | 10 1 1 | 0 0 0 | 0 0 0 | 10 5 5 | 10 0 0 | 0 0 0 | 0 0 0 | 9 0 0 |
Trachea | No. examined | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 9 |
Tracheal Bifurcation | No. examined | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 8 |
Ureters Dilatation | No. examined Total | 0 0 | 0 0 | 0 0 | 1 1 | 0 0 | 0 0 | 0 0 | 0 0 |
Table 7 (cont): Histopathology - group distribution of findings for animals killed after 13 weeks of treatment
Tissue Organs and Findings |
Group/Sex No. of animals | Number of animals affected | |||||||
1M 10 | 2M 10 | 3M 10 | 4M 10 | 1F 10 | 2F 10 | 3F 10 | 4F 9 | ||
Urinary Bladder Infiltrate, Inflammatory Cell | No. examined Minimal Total | 10 1 1 | 0 0 0 | 0 0 0 | 10 0 0 | 9 0 0 | 0 0 0 | 0 0 0 | 9 0 0 |
Uterine Cervix | No. examined | - | - | - | - | 10 | 0 | 0 | 9 |
Uterus | No. examined | - | - | - | - | 10 | 0 | 0 | 9 |
Vagina Diestrus Proestus Metestrus Estrus | No. examined Total Total Total Total | - - - - - | - - - - - | - - - - - | - - - - - | 10 6 3 1 0 | 0 0 0 0 0 | 0 0 0 0 0 | 9 4 2 1 2 |
Table 8: Food consumption – individual values (g/animal/week)
Group /Sex | Cage Number | Week P1 | Week | ||||||||||||
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | |||
1M | 001 | 132 | 125 | 139 | 148 | 149 | 151 | 151 | 147 | 151 | 144 | 146 | 146 | 150 | 142 |
002 | 136 | 127 | 137 | 138 | 136 | 148 | 141 | 142 | 138 | 143 | 139 | 142 | 141 | 131 | |
2M | 003 | 126 | 117 | 133 | 142 | 142 | 145 | 148 | 146 | 149 | 149 | 150 | 149 | 147 | 148 |
004 | 135 | 125 | 137 | 144 | 142 | 140 | 144 | 138 | 142 | 148 | 143 | 150 | 147 | 138 | |
3M | 005 | 126 | 121 | 139 | 143 | 141 | 143 | 146 | 146 | 149 | 146 | 146 | 148 | 148 | 143 |
006 | 133 | 128 | 142 | 149 | 144 | 147 | 146 | 145 | 147 | 150 | 146 | 148 | 148 | 138 | |
4M | 007 | 137 | 129 | 137 | 136 | 137 | 141 | 141 | 134 | 147 | 146 | 145 | 156 | 151 | 135 |
008 | 132 | 128 | 136 | 135 | 133 | 137 | 135 | 133 | 142 | 146 | 140 | 148 | 149 | 132 |
Table 8 (cont): Food consumption – individual values (g/animal/week)
Group /Sex | Cage Number | Week P1 | Week | ||||||||||||
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | |||
1F | 010 | 97 | 105 | 106 | 107 | 107 | 112 | 108 | 107 | 107 | 109 | 107 | 106 | 109 | 104 |
011 | 104 | 105 | 109 | 114 | 111 | 112 | 112 | 114 | 112 | 113 | 111 | 109 | 109 | 105 | |
2F | 012 | 95 | 95 | 102 | 103 | 103 | 106 | 106 | 104 | 104 | 104 | 100 | 101 | 102 | 101 |
013 | 95 | 99 | 105 | 110 | 106 | 110 | 109 | 111 | 112 | 109 | 105 | 107 | 108 | 101 | |
3F | 014 | 108 | 106 | 118 | 117 | 118 | 121 | 122 | 121 | 121 | 120 | 117 | 122 | 117 | 113 |
015 | 108 | 106 | 111 | 116 | 115 | 121 | 118 | 117 | 115 | 116 | 113 | 116 | 114 | 109 | |
4F | 016 | 92 | 104 | 101 | 102 | 99 | 109 | 110 | 104 | 106 | 110 | 108 | 107 | 109 | 98 |
017 | 104 | 100 | 106 | 108 | 108 | 110 | 111 | 109 | 111 | 110 | 110 | 113 | 113 | 102 |
Table 9: Clinical signs – individual observations
Group /Sex | Animal Number | Week of Death |
Phase |
Category |
Observation |
Week(s) observed |
1M | 0004 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 10-14 |
0006 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface |
7-14 9-14 | |
0008 | 14 | Treatment | Coat Skin abrasion Staining | Hair loss, Dorsal surface Dry, Lower dorsal surface Abnormal color, Brown, Dorsal surface | 10-14 10-13 10-14 | |
2M | 0012 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 8-14 |
0016 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Upper ventral surface | 8-14 12-14 | |
0017 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Upper dorsal surface | 12-14 9-11 | |
0018 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 13-14 | |
0020 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 13-14 | |
3M | 0024 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Upper ventral surface | 9-14 13-14 |
0026 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 10-14 | |
0027 | 14 | Treatment | Skin abrasion Staining | Wet, Hindlimb digit(s) Abnormal color, Brown, Upper dorsal surface | 3 9-14 | |
0029 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 9-14 | |
0030 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 13-14 | |
4M | 0032 | 14 | Treatment | Coat Skin abrasion | Hair loss, Dorsal surface Dry, Tail | 1-2 8 |
0035 | 14 | Treatment | Coat Skin abrasion | Hair loss, Dorsal surface Wet, Hindlimb digit(s) | 8-14 12-13 | |
0036 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 10-14 | |
0037 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 9-14 | |
0040 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 9-14 |
Only animals with observations are presented
Table 9 (cont): Clinical signs – individual observations
Group /Sex | Animal Number | Week of Death |
Phase |
Category |
Observation |
Week(s) observed |
1F | 0101 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Ventral surface | 8-14 9-13 |
0102 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 7-14 7-14 | |
0103 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 7-14 | |
0104 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 8-14 8-14 | |
0105 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Ventral surface | 8-14 10-14 | |
0106 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 7-14 8-14 | |
0108 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Upper dorsal surface | 10-14 9 | |
0110 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Upper dorsal surface | 12-14 9-11 | |
2F | 0111 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 8-14 8-14 |
0112 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 7-14 6, 8-14 | |
0113 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Ventral surface | 10-14 9-14 | |
0115 | 14 | Treatment | Coat Staining | Hair loss, Dorsal surface Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 12-14 12-14 9-14 | |
0116 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 13-14 | |
0118 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Ventral surface | 8-14 10-14 | |
0119 | 14 | Treatment | Staining | Abnormal color, Brown, Lower ventral surface Abnormal color, Brown, Upper dorsal surface | 11-14 9-14 | |
0120 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 9-14 | |
3F | 0121 | 14 | Treatment | Staining | Abnormal color, Brown, Ventral surface | 9-14 |
0122 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 6-14 | |
0123 | 14 | Treatment | Staining | Abnormal color, Brown, Ventral surface | 9-14 | |
0124 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Whole body | 13-14 5-8 9-12 | |
0125 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 7-14 | |
0127 | 14 | Treatment | Staining | Abnormal color, Brown, Ventral surface | 10-14 | |
0128 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Ventral surface | 13-14 9-12 13-14 | |
0130 | 14 | Treatment | Staining | Abnormal color, Brown, Dorsal surface | 8-14 | |
4F | 0131 | 14 | Treatment | Coat Staining | Hair loss, Dorsal surface Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Ventral surface | 1-2 11-14 9-14 |
0132 | 14 | Treatment | Coat | Hair loss, Dorsal surface | 8-14 | |
0134DD | 7 | Treatment | Reason for dispatch | Found dead in a restraint tube | 7 | |
0135 | 14 | Treatment | Coat | Hair loss, Ventral surface | 3 | |
0136 | 14 | Treatment | Coat Staining | Hair loss, Dorsal surface Abnormal color, Brown, Upper dorsal surface Abnormal color, Brown, Ventral surface | 13-14 9-14 | |
0137 | 14 | Treatment | Coat | Hair loss, Head | 4-5 | |
0138 | 14 | Treatment | Staining | Abnormal color, Brown, Ventral surface | 10-14 | |
0139 | 14 | Treatment | Staining | Abnormal color, Brown, Upper dorsal surface | 9-14 | |
0140 | 14 | Treatment | Coat Staining | Hair loss, Dorsal surface Abnormal color, Brown, Dorsal surface | 9-14 13-14 |
DD – Found dead after dosing
Only animals with observations are presented
Applicant's summary and conclusion
- Conclusions:
- Exposure of Han Wistar rats to ethylal by inhalation for 13 weeks (6 hours per day, 5 days per week) at achieved aerosol concentrations of 505, 1520 or 3860 ppm was well tolerated. Treatment related clinical signs of unsteady gait and/or swaying, decreased activity, salivation and wet rales were intermittent, transient and considered not to be adverse at these concentrations.
Exposure of the highest dose group to an above target ethylal concentration for 3 days in Week 7 (3 day mean concentration 4920 ppm, presented for information only), resulted in a single unscheduled death and an increase in the number and type of clinical signs evident for other rats in this group. These signs were consistent with exposure at 4810 ppm on a previous dose range finding study.
Pathological changes in the thyroid glands and liver in males exposed to 3860 ppm are considered rodent-specific, adaptive and non-adverse.
The No Adverse Effect Level was considered to be 3860 ppm. - Executive summary:
The purpose of this study was to assess the systemic toxic potential of ethylal, an industrial chemical, in a 13 week inhalation study in Han Wistar rats.
The study design was as follows:Group
Treatment
Target exposure level (ppm)
Number of animals
Male
Female
1
Air control
0
10
10
2
Ethylal
500
10
10
3
Ethylal
1500
10
10
4
Ethylal
4000
10
10
Animals received the control, air or the test item, ethylal by inhalation for 13 weeks (6 hours daily exposure for 5 days per week).
During the study, clinical condition, body weight, food consumption, ophthalmoscopy, hematology (peripheral blood), blood chemistry, organ weight, bronchoalveolar lavage, macropathology and histopathology investigations were undertaken.
Results
Summary data are presented below:Group
Atmosphere Concentration (ppm)
Target
Achieved
1
0
-
2
500
505
3
1500
1520
4
4000
3860
The mean achieved aerosol concentrations were 101, 101 and 97% of target for Groups 2, 3 and 4, respectively, with little daily variation.
In error, a draining valve (used as part of once weekly cleaning procedures) on the Group 4 exposure chamber, was not closed prior to the start of the first daily exposure in Week 7. The error was detected during exposure on Week.Day 7.4. Failure to close this valve is considered to have resulted in reduced efficiency of the chamber aerosol extract and thus higher achieved aerosol concentrations on these days. Higher than target aerosol concentrations were measured for all chamber aerosol samples collected from Group 4 on Week.Days 7.1, 7.2 and 7.3 and the first sample collected on Week.Day 7.4. Mean achieved aerosol concentrations for Group 4 on Week.Days 7.1, 7.2 and 7.3 were 127%, 112%, 131% of target respectively (3 day mean concentration 4920 ppm). Due to the failure of the analytical standard comparison, the aerosol concentrations from Week.Days 7.1, 7.2 and 7.3 (together with the results from Week.Day 6.1 to 6.5 and Week.Day 7.4 to 7.5) are reported for information purposes only and are excluded from the overall group mean.Animal 134, a female receiving the highest exposure level of ethylal was found dead at the end of exposure on Week.Day 7.1. On Week.Days 7.1 to 7.3, clinical signs evident for rats receiving the highest ethylal exposure level were more marked than those seen after other exposures. Signs seen after exposure, for a proportion of rats, included locomotor and responsiveness effects (decreased activity, unsteady gait, unresponsiveness, flattened posture) and breathing effects (slow and/or shallow breathing and wet rales). Animal 138 (a high exposure level female) was removed from exposure on Week.Day 7.2, after approximately 4.5 hours, due to welfare concerns. Clinical signs evident for this animal included abnormal gait (unsteady and elevated), hunched posture, shallow breathing, decreased activity and pale ears; but it had recovered by the end of the working day.
Unsteady gait was occasionally observed for rats receiving ethylal at all exposure levels. In addition, swaying, decreased activity, lacrimation and wet rales were occasionally observed for a proportion of rats exposed to 3860 ppm. These signs were observed on return to home cage and resolved by the end of the working day.
Pathological examination revealed follicular cell hypertrophy in the thyroid of five males exposed to 3860 ppm and hypertrophy of the centrilobular hepatocytes of the liver in four of the same males. These findings are considered rodent-specific, adaptive and non-adverse.
Conclusion
Exposure of Han Wistar rats to ethylal by inhalation for 13 weeks (6 hours per day, 5 days per week) at achieved aerosol concentrations of 505, 1520 or 3860 ppm was well tolerated.
Treatment related clinical signs of unsteady gait and/or swaying, decreased activity, salivation and wet rales were intermittent, transient and considered not to be adverse at these concentrations.
Exposure of the highest dose group to an above target ethylal concentration for 3 days in Week 7 (3 day mean concentration 4920 ppm, presented for information only), resulted in a single unscheduled death and an increase in the number and type of clinical signs evident for other rats in this group. These signs were consistent with exposure at 4810 ppm on a previous dose range finding study.
Pathological changes in the thyroid glands and liver in males exposed to 3860 ppm are considered rodent-specific, adaptive and non-adverse.
The No Adverse Effect Level was considered to be 3860 ppm.
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