Registration Dossier
Registration Dossier
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EC number: 701-301-6 | CAS number: -
Endpoint summary
Administrative data
Description of key information
Key study: Test method OECD 422. GLP study. The oral 28d-NOAEL in rats was determined to be 300 mg/kg bw/day since no adverse effects were observed at the highest dose tested.
Key study: Test method OECD 408 and EU Method B.26. GLP study. The NOAEL for the subchronic repeated dose toxicity of the test item in rats by oral route was determined to be 250 mg/kg bw/day.
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 26.08.2020-24.03-2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Deviations:
- yes
- Remarks:
- During the experiment the humidity dropped below 30% or exceed 70% (22.1 – 92.9%) a few times as well as the temperature dropped below 20°C (15.9 – 22.5°C). This changes were temporary and did not influence on the study course.
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- (Cmdb: Wi; outbred)
- Details on species / strain selection:
- The animal species (Rattus norvegicus) is recognized by OECD guideline No. 408 as a recommended test system for repeated dose oral toxicity study in rodents
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: The husbandry of laboratory animals of the Experimental Medicine Centre at the Medical University in Białystok kept behind a breeding barrier (unit number in the register of units entitled to husbandry of laboratory animals: 043)
- Age at study initiation: 6-7 weeks
- Weight at study initiation:
Group 0: males 210.1g (193g – 236g); females 155.8g (143g – 177g)
Group 1: males 210.2g (190g – 229g); females 155.8g (146g – 175g)
Group 2: males 210.1g (194g – 233g); females 155.8g (144g – 176g)
Group 3: males 210.1g (192g – 227g); females 155.8g (142g – 168g)
Group 0 SAT: males 197.3g (180g – 218g); females 160.9g (151g – 175g)
Group 3 SAT: males 197.4g (188g – 218g); females 160.9g (151g – 172g)
- Fasting period before study: no
- Housing: The animals were kept in air-conditioned cages (58x37x21) with a plastic bottom and covered with wire-bar lids and UV-sterilized wood chips were used as bedding.
- Diet: Altromin 1324 P TPF” (Phytoestrogen-poor, Total Pathogen Free) standard granulated fodder produced by Altromin Spezialfutter GmbH & Co. KG, Lage, Germany (batch numbers: 5/20, 6/20)
- Water: Drinking tap water ad libitum
- Acclimation period: 5 days
DETAILS OF FOOD AND WATER QUALITY:
Ad libitum access to the “Altromin 1324 P TPF” (Phytoestrogen-poor, Total Pathogen Free) standard granulated fodder produced by Altromin Spezialfutter GmbH & Co. KG, Lage, Germany (batch numbers: 5/20, 6/20) (Appendix No. 8) and drinking tap water.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15.9 – 22.5°C
- Humidity (%): 22.1 – 92.9%
- Air changes (per hr): about 15 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/ 12 hours dark - Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Remarks:
- Batch number / expiry date: 20259 / 05.2021, 20342 / 06.2021; Storage / stability: stored at room temperature (15-25 °C); Manufacturer: Oleofarm Sp. z o.o., ul. Mokronoska 8, 52-407 Wrocław; Packaging: brown glass bottles, volume 0.5 L
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
Dose of 62.5 mg/kg b.w./day: There was 12.5 mg VL3 in 1 mL (13.6 mg in 1g) of corn oil solution.
Dose of 250 mg/kg b.w./day: There was 50 mg VL3 in 1 mL (54.4 mg in 1g) of corn oil solution.
Dose of 1000 mg/kg b.w./day: There was 200 mg VL3 in 1 mL (217.7 mg in 1g) of corn oil solution.
VEHICLE
- Justification for use and choice of vehicle (if other than water): Test item is soluble in inert organic solvents like corn oil
- Concentration in vehicle: 12.5, 50 and 200 mg/mL
- Amount of vehicle (if gavage): The volume of the dose formulation (X) administered to each animal was calculated according to the formula: X= 0.5 mL x animal body weight [g]/100 g.
- Lot/batch no. (if required): 20259
- Purity: not specified - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Equipment:
Gas Chromatograph (6890N network GC System, Agilent)
Usual laboratory material (flasks, pipettes, balances)
Analytical method:
The concentration of VL3 was chemically determined with gas chromatographic method with FID. The validation was involved the assessment of the range of linearity of the analytical graph, specificity, precision (repeatability), recovery, and limits of quantification and detection. The test item was used as the standard.
Method validation:
The method was validated at two fortification levels (5.0 mg/g and 50 mg/g) for corn oil. For each fortification level and matrix, five replicates were prepared and analysed. Additionally, two replicates of unfortified samples were analysed (control samples).
The method validation conducted to prove the validity of the applied method showed that the method was suitable to determine VL3 in corn oil. The mean recoveries for matrix ranged from 96.6% to 100.6%. Therefore, the obtained recovery values fulfill the legal requirement for recovery values (SANCO/3029/99 rev. 4).
For analyte, the relative standard deviations (RSD) were below 10% and no significant peak occurred at the retention time, so absence of interference was confirmed.
Storage Stability of Samples:
The experiments demonstrate that VL3 was not stable in corn oil over a time period of 4 days, when stored at ambient temperature (day 0 result of recovery is 98.6%, day 4 result of recovery is 10.7% (result below LOD i.e. 1.5 mg/g).
Chemical analysis during the study:
Test item formulations were transferred thrice for chemical analysis on: 28.08.2020, 08.10.2020 and 23.11.2020. The mean recovery values for test samples ranged from 92.4 to 103.6%. All results were in the range from 80 to 120%, therefore the acceptance criteria for test samples for toxicological study on animals was met. The results of the determination of VL3 in test samples are presented in Table 3. - Duration of treatment / exposure:
- 90 days
- Frequency of treatment:
- Animals from all groups were treated with the test item/control item once daily, seven days a week (the first dosing day = day 0).
- Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- GROUP 0 AND SAT0
- Dose / conc.:
- 62.5 mg/kg bw/day (nominal)
- Remarks:
- GROUP 1
- Dose / conc.:
- 250 mg/kg bw/day (nominal)
- Remarks:
- GROUP 2
- Dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- Remarks:
- GROUP 3 AND SAT 3
- No. of animals per sex per dose:
- 10 males and 10 females in each group
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: Dose levels as well as vehicle were established based on existing data according to OECD guideline No. 422. In this guideline, the doses were: 33.3 mg/kg b.w., 100 mg/kg b.w. and 300 mg/kg b.w. where no test item- related lesions in parental animals were stated. Dose of 1000 mg/kg b.w./day is the accepted dose by OECD to be the limit dose up to which adverse effects should be checked. Due to longer time of exposition, higher level of doses with 4-fold intervals in current study results of the study should allow for estimating the NOAEL or LOAEL.
- Rationale for animal assignment (if not random): The animals were randomly divided according to stratification by body weight into four experimental groups and two satellite groups.
- Fasting period before blood sampling for clinical biochemistry: 15 hours
- Post-exposure recovery period in satellite groups: 21 days - Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day usually at the beginning and end of each day
- Cage side observations checked: mortality and morbidity
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: before the test item / vehicle administration on day 0 and then once a week.
BODY WEIGHT: Yes
- Time schedule for examinations: prior to the beginning of the experiment (day 0) and then twice a week during the experiment and on the last day of experiment
FOOD CONSUMPTION: Food consumption was measured, although it is not a feeding study.
Food in cages was weighted to determine food intake once a week during the experiment.
FOOD EFFICIENCY: No
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: before the study begins and at the end of the study (on the 89th day for groups 0, 1, 2 and 3 or on the 110th day for groups 0 SAT and 3 SAT). In case of the satellite groups, it also took place after the end of the test item / vehicle administration (on the 89th day)
- Dose groups that were examined: 0, 1, 2, 3, 0 SAT and 3 SAT
HAEMATOLOGY: Yes
- Time schedule for collection of blood: at main study termination (90th day) and on satellite animals on the day of recovery sacrifice (111th day)
- Anaesthetic used for blood collection: Yes, xylazine-ketamine mixture at a dose of: 10 mg/kg b.w. of xylazine and 100 mg/kg b.w. of ketamine
- Animals fasted: Yes
- How many animals: All of them
- Parameters examined: leukocyte count, erythrocyte count, thrombocyte count, level of hemoglobin, hematocrit value. The morphological results contain the calculated erythrocyte indices such as: mean corpuscular volume (MCV), mean corpuscular hemoglobin weight (MCH), mean corpuscular hemoglobin concentration (MCHC).
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at main study termination (90th day) and on satellite animals on the day of recovery sacrifice (111th day)
- Animals fasted: Yes
- How many animals: All of them
- Parameters examined: total protein, albumin, level of globulin, total cholesterol, urea nitrogen, creatinine, glucose, bilirubin, calcium, sodium, potassium, chlorides, inorganic phosphorus, high - density lipoproteins (HDL) and low - density lipoproteins (LDL).
- Enzymatic examinations: The activity of the following enzymes were determined in serum: − aspartate aminotransferase (AST).
− alanine aminotransferase (ALT).
− alkaline phosphatase (AP).
− gamma - glutamyl transferase (GGT).
SERUM HORMONES: Yes
- Time of blood sample collection: at a comparable time of the day
- Animals fasted: Not specified
- How many animals: All of them
- The level of thyroxine (T4), triiodothyronine (T3) in serum was determined ELISA method using Calbiotech kits: Mouse/Rat Triiodothyronine (T3) ELISA , Mouse/Rat Thyroxine (T4) ELISA. The level of thyroid stimulating hormone (TSH) in serum was determined ELISA. All hormones determinations were made using FLUOstar Omega reader.
URINALYSIS: Yes
- Time schedule for collection of urine: at main study termination (90th day) and on satellite animals on the day of recovery sacrifice (111th day)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined: volume of urine (mL), urine colour, specific gravity, pH, protein (g/L), glucose (mmol/L), ketone bodies (mmol/L), bilirubin (qualitative test), blood (Ery/μL), urobilinogen (μmol/L), leukocyte count (leu/μL) and nitrites (qualitative test).
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Groups 0, 1, 2 and 3: during the thirteenth week of the experiment (between the 85th and 89th day). Satellite groups: during the thirteenth week of the experiment - measurement 1 (between the 85th and 89th day) and at the end of the sixteenth week of the experiment - measurement 2 (between 104th and 110th day)
- Dose groups that were examined: 0, 1, 2, 3, 0 SAT and 3 SAT
- Battery of functions tested: open field observations, evaluation of responses to simuli, measurement of the fore- and hindlimb grip strength, evaluation of locomotor activity.
IMMUNOLOGY: No
OTHER:
Bone marrow examinations: Bone marrow was obtained from a dissected femur after animal euthanasia. The collected bone marrow pellet was transferred to an eppendorf tube containing 50μl FBS. After a few minutes and gently mixing, a smear was made. The slides were stained May-Grunwald-Giemsa method and were qualitatively and quantitatively evaluated through the determination of the number of individual nuclear cells per 1000 test cells.
The number of following cells were determined:
- within the erythrocyte system,
- within the leukocyte system,
- within the range of different cells: lymphocytes, monocytes, plasmocytes, megakaryocytes and other cells
Bone marrow was assessed in males and females from the control groups i.e. group 0 and recovery group 0 SAT and highest dosage group i.e. group 3 and recovery group 3 SAT. Microscopic analysis was not extended to groups 1 and 2, as no treatment-related changes were found in groups 3 and 3 SAT.
Coagulation examinations: Prothrombin time (PT) and activated partial thromboplastin time (APTT) was determined in plasma collected in tubes containing anticoagulant sodium citrate 3.2% at a 1:9 ratio (sodium citrate:blood). PT and APTT evaluation was based on the changes of the optical density measured chronometrically with the use of the Coag Chrom 3003 coagulometer and BioKsel reagents. - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
All animals which survived the experiment after the collection of blood from the heart for the clinical-chemical examinations, and were euthanized with a xylazine-ketamine mixture at a dose of 20 mg/kg b.w. of xylazine and 200 mg/kg b.w. of ketamine, as well as one female euthanized for humanitarian reasons during the experiment, were subjected to post-mortem examinations. Gross examination which was carried out on all animals participating in the experiment, consisted of thorough observations of the external body surface and all orifices, all natural apertures as well as the cranial, thoracic and abdominal cavities with their content. From each animal from which the blood was taken for clinical-chemical tests, one femur was collected for bone marrow examination.
From all females involved in the experiment during gross examination vaginal smears were collected and estrus cycle of all females was determinated.
After the end of experiment, absolute weights of particular internal organs of all surviving animals were determined. The organs were as follows: brain, thymus, heart, liver, spleen, kidneys, adrenal glands, testes, epididymides, uterus, ovaries, thyroid with parathyroid, prostate with the seminal vesicles and coagulating glands, pituitary gland.
The relative weight of internal organs were calculated on the basis of the absolute organ weight with reference to the live animals’ body weight after fasting.
HISTOPATHOLOGY: Yes
The following organs and tissues of all animals were preserved for histopathological examinations: brain (representative regions including cerebrum, cerebellum and medulla/pons), pituitary, spinal cord (cervical, mid-thoracic and lumbar), eyes with the optic nerve (if changes were observed during ophthalmological examinations), mandibular salivary glands, stomach, small and large intestines (including Peyer's patches if macroscopical visible) i.e.: duodenum, jejunum, ileum, cecum, colon, and rectum; liver, pancreas, kidneys, adrenal glands, urinary bladder, spleen, heart, aorta, thymus, lymph nodes (mandibular and mesenteric), thyroid with parathyroid, oesophagus, trachea, lungs, ovaries, uterus with cervix, vagina, vaginal smears, testes, epididymides, prostate with seminal vesicles and coagulating glands, skeletal muscle including peripheral nerve (sciatic or tibial), femur, skin, mammary gland and all tissues showing abnormality. Collected organs and tissues were fixed in a 10% solution of formalin, except the testes and epididymides which were fixed in modified Davidson’s fluid. Bones were subjected to a decalcification.
Preserved samples of all collected organs and tissues were embedded in paraffin, stained with hematoxylin and eosin, and evaluated under a light microscope.
Due to no observed changes in ophthalmological examinations in all surviving animals, histopathological examination of eyes with optic nerves was not conducted. It was conducted only in the female euthanized for humanitarian reasons during the experiment, due to lack of ophthalmological examination.
Full histopathological examination was performed on all preserved organs and tissues of all animals from the control and high dose groups (group 0 and group 3). All gross lesions were examined.
Due to occurrence of the treatment-related lesions in urinary bladder and kidneys of animals from high dose group (group 3), histopathological examination of these organs from other dosage groups, as well as from satellite groups, was conducted. - Statistics:
- The results obtained in the study were subjected to statistical analysis, using following programs: Excel 2007 and STATISTICA 10.0 PL.
The results are presented in tables in the form of average values and standard deviation.
The treated groups i.e. groups 1, 2, and 3 were compared to the control group (group 0).
The course of the statistical analysis (group 0, 1, 2, 3) was as follows:
- first of all, the normality of the distribution was examined using the Shapiro-Wilk test and the homogeneity of variance by the Brown-Forsythe test,
- results which were characterized by normal distribution and homogeneous variances were compiled using a one-way analysis of variance followed by Dunnett's test,
- in the absence of normality of distribution or non-homogeneous variances, the nonparametric Kruskal-Wallis test was used, followed by Dunnett's test.
The treated satellite groups i.e. 3 SAT were compared to the satellite control groups (group 0 SAT). The following statistical tests were used to assess intergroup relationships for groups 0 SAT and 3 SAT:
- first of all, the normality of the distribution was examined using the Shapiro-Wilk test and the homogeneity of variance by the Brown-Forsythe test,
- results which were characterized by normal distribution and homogeneous variances were compiled using the Student’s,
- parameters characterized by non-normal distribution were compiled using Mann-Whitney U test,
- parameters characterized by normal distribution and non-homogenous variance were compiled by Cochrane-Cox test.
Ordinal variables (part of urine parameters) were analyzed using Kruskal-Wallis test followed by Dunnett's test (main groups) or Mann-Whitney U-test (satellite groups).
The weights of internal organs are presented in tables as absolute values as well as relative values (with reference to the body weight of live animals and to weight of brain), in the form of average values and standard deviation.
The obtained results were analyzed statistically at p ≤ 0.05 - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Group 0:
− 2 males and 1 female showed thinning of coat on one or both cheeks at some point during the experiment and the female posteriorly showed alopecia on the left cheek.
Group 1:
− 1 male showed thinning of coat on the right cheek and 1 female showed thinning of coat on both sides of neck and then alopecia on both sides of neck.
Group 3:
− 1 male showed thinning of coat on both forearms.
− 2 males: showed thinning of coat on both cheeks and one of them alopecia on the left cheek.
− 1 female showed thinning of coat on both sides of neck and then alopecia on both sides of neck
− 3 females showed thinning of coat on one or both cheeks.
− 1 female showed thinning of coat on both forearms.
Group 0 SAT:
− 1 female showed thinning of coat on the right thoracic limb, then alopecia on the right thoracic limb and on the right side of abdomen and finally then alopecia on the right thoracic limb, on the right side of abdomen and the left forearm.
Group 3 SAT:
− 1 male showed alopecia on both forearms.
− 1 male showed thinning of coat on both sides of loins.
− 1 female showed thinning of coat on the right cheek.
− 1 female showed dejection, distinct decrease of locomotor activity, no intake of food and water. Later, this animal lied on its abdomen motionless, with no reaction to external stimuli and catching and no intake of food and water, pallor of the skin and irises, bristled coat and hypothermia. It was euthanized. - Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- MALES:
- Significant increase of average food intake was found in the males of groups 1 and 3 (12th week), in the group 2 (10th week), and group 3 SAT (6th, 8th, 12th, 13th, 14th and 15th week).
- Significant decrease of average food intake was found in the males of group 3 SAT(2nd week).
FEMALES:
- Significant increase of average food intake was found in the females of group 1 (4th week), and group 3 SAT (15th week).
- Significant decrease of average food intake was found in the females of group 3 SAT in (1st week).
These effects are considered to be accidental because lack of confirmation in body weight measurements. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- MALES:
- Decrease in the MCV, MCH and reticulocytes values in group 3.
FEMALES:
- Decrease in the erythrocytes number and MCHC value and increase in the MCV value and number of leukocytesin group 3 SAT. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- -Increase in the potassium concentration in group 3 SAT.
- Endocrine findings:
- no effects observed
- Urinalysis findings:
- effects observed, treatment-related
- Description (incidence and severity):
- MALES:
-Significant decrease in pH value and increase in the number of transitional epithelia and leukocytes in group 3.
FEMALES:
- Increase in the number of transitional epithelia and leukocytes in group 3 and decrease in the amount of ammonium- magnesium phosphates in group 3 SAT. - Behaviour (functional findings):
- effects observed, treatment-related
- Description (incidence and severity):
- - Significant decrease of the horizontal locomotor activity and vertical locomotor activity in males from group 3 SAT, probable related to the increase of potassium levels.
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- - Significant increase of weight in males from group 3 in kidneys.
- Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- - Significant amount of males with thickened wall in urinary bladder from group 3 (4) compared to the rest of the main groups (0)
-Significant amount of females with thickened wall in urinary bladder from group 3 (6) compared to the rest of the main groups (0) - Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- MALES:
- Group 3: Significant amount with moderate or severe papillary hyperplasia of urothelium in urinary bladder and left kidney, and hyperplasia of papilla epithelium effects in both kidneys.
- Group 3: Significant amount with minimal, slight or moderate renal pelvis and papillary hyperplasia of urothelium effects in both kidneys.
- Group 3 and 3SAT: Significant amount of basophilic (renal) tubules was observed in both kidneys.
FEMALES:
- Group 3: Significant amount with severe papillary hyperplasia of urothelium in urinary bladder
- Group 3: Significant amount with slight, moderate or severe papillary hyperplasia of urothelium and hyperplasia of epithelium effects in both kidneys.
- Group 3: Significant amount with minimal, slight or moderate renal pelvis and papillary hyperplasia of urothelium effects in both kidneys. - Histopathological findings: neoplastic:
- not examined
- Details on results:
- ENZYMATIC EXAMINATIONS:
-Increase in the AST and ALT activity in males in group 3 SAT compared to the control group 0 SAT. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 250 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical biochemistry
- clinical signs
- histopathology: non-neoplastic
- organ weights and organ / body weight ratios
- urinalysis
- Key result
- Critical effects observed:
- yes
- Lowest effective dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- System:
- urinary
- Organ:
- bladder
- kidney
- Treatment related:
- yes
- Dose response relationship:
- yes
- Relevant for humans:
- not specified
- Conclusions:
- The NOAEL for the sub-chronic repeated dose toxicity of the test item in rats by oral route was determined to be 250 mg/kg bw/day.
- Executive summary:
A 90-day repeated dose toxicity study was conducted with the test item according to OECD Guideline 408 and EU Method B.26 (GLP study). The test item was exposed to 120 Wistar rats (60 males, 60 females), which were divided into four groups main groups, one control and three treated groups, and two satellite groups. There were 10 males and 10 females in each group. The doses of the test item were 62.5, 250 and 1000 mg/kg b.w./day. During the study, clinical signs, mortality, body weights, and food intake were recorded. Moreover, ophthalmic examinations and behavioural studies were conducted. Behavioural studies involved observations of animal behaviour in an open field, responses to stimuli, locomotor activity, and fore- and hindlimb grip strength. At the end of the experiment, all animals were subjected to hematological examinations of peripheral blood and bone marrow, coagulation examinations, biochemical and enzymatic examinations of serum, general urinalyses, examinations of urine sediment and hormone examinations. After the end of experiment, absolute weights of particular internal organs of all surviving animals were determined. An extensive amount of organs and tissues were preserved for histopathological examinations.
On the basis of the results of the clinical examinations, the test item induced a decrease in the vertical locomotor activity (males group 3 SAT), which may be related to an increase in the amount of potassium, as found in the clinical-chemical examinations. In the post-mortem examination, it was observed papillary hyperplasia of urothelium in urinary bladder and kidneys as well as hyperplasia of papilla epithelium in kidneys - in males and females of group 3, a significant increase of relative weight of kidneys - in males of group 3 and the presence of basophilic tubules - in males of group group 3 and 3SAT.
It may be concluded that the test item related changes concerned especially kidneys and urinary bladder at high dose 1000 mg/kg b.w/day, which were observed in males and females. Despite the reversibility of some of them, changes resulted in an adverse effect on animal well-being, morphology and physiology during the study and also after recovery period. Based on this, the No Observed Adverse Effect Level (NOAEL) of the test item in rats of this study was determined to be 250 mg/kg b.w./day for males and females.
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 24.03.2016 to 22.01.2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Analytical purity: 93%
- Expiry date: 06.03.2018 - Species:
- rat
- Strain:
- Wistar
- Remarks:
- Cmdb:WI; outbred
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: from the husbandry of laboratory animals of the Experimental Medicine Centre at the Medical University in Białystok,
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 13-14 weeks old.
- Weight at study initiation: average body weights:
- group 0: males 403.50 g; females 243.75 g
- group 1: males 403.50 g females 243.92 g
- group 2: males 403.40 g; females 243.75 g
- group 3: males 403.50 g; females 243.92 g
- group 0 SAT: males 404.00 g; females 244.90 g
- group 3 SAT: males 404.10 g; females 245.10 g.
- Fasting period before study:
- Housing: Air-conditioned rooms. The animals were kept in plastic cages covered with wire bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height). During the main experiment, there were 2 or 3 animals/cage. Each sex was kept separately.
- Diet (e.g. ad libitum): ad libitum access,
- Water (e.g. ad libitum): ad libitum access.
- Acclimation period: All animals were quarantined and observed for at least 5 days.
DETAILS OF FOOD AND WATER QUALITY: Food: Altromin 1324 P TPF (Phytoestrogen Deficient, Total Pathogen Free) standard granulated fodder produced by Altromin Spezialfutter GmbH & Co. KG, Lage, Germany, batch number: 1/17 -200418/0453. A certificate of analysis is included in the report.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25ºC
- Humidity (%): 43 - 81%
- Air changes (per hr): 13-18 times/h
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark
IN-LIFE DATES: From: Experimental starting date 13.06.2017 To: Experimental completion date 22.08.2017 - Route of administration:
- oral: gavage
- Details on route of administration:
- The test item and the vehicle control were administered to animals once a day for seven days by gavage, in a constant volume of 0.5 mL/100 g b.w. The amounts of the solution of the test item were adjusted to animals’ body weights determined on body weight measurement days.
- Vehicle:
- corn oil
- Details on oral exposure:
- - PREPARATION OF DOSING SOLUTIONS:
During the main study solutions of the test item in a vehicle (corn oil) were prepared once a week. Prepared solutions were stored in a fridge (temperature 4-8 ºC). During administration, solutions were kept at room temperature and they were thoroughly mixed.
- VEHICLE
- Justification for use and choice of vehicle (if other than water): most suitable vehicle, as the test item hydrolyses in water.
- Concentration in vehicle: 6.66 mg VL3/1 mL solution, 20 mg VL3/1 mL solution and 60 mg VL3/1 mL solution
- Amount of vehicle (if gavage): the total volume administered was 0.5mL/100 g b.w. (including test item)
- Lot/batch no. (if required): L5294 (Supplier BASSO FEDELE & FIGLI srl, ITALY)
- Purity: 100 % corn oil - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- It was conducted by BLIRT S.A. Laboratory, Trzy Lipy 3/1.38, 80-172 Gdańsk, Poland, which holds the Good Laboratory Practice Certificate GLP. Solutions of the test item in a vehicle (corn oil) were prepared once a week. Two samples (2 x 2 ml) were collected from each batch of prepared solutions. One of them was frozen and sent in a pack of dry ice to BLIRT S.A. (27 samples in 9 batches) for analysis and the other one (spare sample) was archived in the freezer (-15 - -20°C).
TECHNIQUE AND TEST METHOD: Gas chromatography coupled with flame ionization detector (FID).
TEST PARAMETERS: Chromatographic column: ZB-5MS, 30 m x 0.25 mm x 0.25 μm, catalog no.: 7HG-G010-11, serial no.: 221879, Phenomenex.
Temperature gradient: 175ºC (Injection port), 280ºC (detector), 25ºC/min column rate., Column flow: 0.69 mL/min, Carrier gas: helium, Injection volume: 1 μL
Split ratio: 33.3 mg/kg b. w. (6.66 mg/mL of VL3 in corn oil) and 100 mg/kg b. w. (20 mg/mL of VL3 in corn oil): 1:50, 300 mg/kg b. w. (60 mg/mL of VL3 in corn oil): 1:100.
ACCEPTANCE CRITERIA: The relative standard deviation, RSD [%], of the VL3 content for 3 results for 33.3 mg/kg b. w. (6.66 mg/mL of VL3 in corn oil) dose is 6.67 %, so it is ≤ 10.0 %. The relative standard deviation, RSD [%], of the VL3 content for 3 results for 100 mg/kg b. w. (20 mg/mL of VL3 in corn oil) dose is 9.91 %, so it is ≤ 10.0 %. The relative standard deviation, RSD [%], of the VL3 content for 3 results for 300 mg/kg b. w. (60 mg/mL of VL3 in corn oil) dose is 0.48 %, so it is ≤ 10.0 %. The acceptance criteria are fulfilled.
QUANTITATIVE AND QUALITATIVE DETERMINATION: All average VL3 content in study samples were at the expected level. - Duration of treatment / exposure:
- The test item/vehicle control was administered to males for 28 days.
Females were dosed throughout the study. This included 2 weeks prior to mating, the variable time to conception, the duration of pregnancy, and at least 13 days after delivery (51 – 59 days).
Males from groups 0 SAT and 3 SAT were treated for 28 days, females from groups 0 SAT and 3 SAT were treated for 53 days. Post treatment, the satellite groups were observed for 14 days. - Frequency of treatment:
- Daily
- Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- Only receiving corn oil (vehicle control)
- Dose / conc.:
- 33.3 mg/kg bw/day (nominal)
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- Group 0, 1, 2, 3: 10 males, 12 females
Satellite groups: 10 males, 10 females - Control animals:
- yes, concurrent vehicle
- other: two satellite groups
- Details on study design:
- - Dose selection rationale:
Two Dose range-finding studies were conducted.
First Dose range-finding study: administered 250, 500 and 1000 mg/kg bw/d (n= 5animals/sex/group) during 7 days. In haematological examinations, statistically significant decrease in the number of leukocytes in males from group 1 and females from group 1 and 3 were stated. Biochemical examinations revealed following statistically significant changes between the treated and the control groups: the decreased total protein concentration in males from group 1, the decreased albumin concentration in males from group 1, the decreased globulin concentration in males from group 1, the decreased calcium concentration in males from group 1, the decreased creatinine concentration in females from group 3.
Based on these results, a second Dose range-finding study was conducted: administered 31.3, 65.5, 125 mg/kg b.w./day for 7 days.It was not observed treatment-related changes.
Based on both dose range finding studies, the doses of 33.3, 100 and 300 mg/kg b.w./day were selected for the main study.
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: control and highest dose group in order to evaluate the reversibility, stability, or delay in the onset of possible harmful effects.
- Post-exposure recovery period in satellite groups: 14 days - Positive control:
- Not conducted
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day or once a day (on days off)
- Cage side observations were: General condition of the animals, i.e. the observation of all animals for morbidity and mortality was conducted.
DETAILED CLINICAL OBSERVATIONS: Yes, evaluation of skin, fur, eye and mucosa changes, the respiratory, circulatory, and autonomous and central nervous systems, somatic activity, and behaviour .
- Time schedule: before the experiment and then once a week.
BODY WEIGHT: Yes
- Time schedule for examinations: twice a week during the entire experiment.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, once a week
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes (g/100 g b.w./day)
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: the day before the introduction of the animals to the experiment and one day before euthanasia.
- Dose groups that were examined: all adult animals
HAEMATOLOGY: Yes
- Time schedule for collection of blood: at termination of the experiment, blood was collected for clinical-chemical examinations.
- Anaesthetic used for blood collection: Yes, anesthetized with a mixture of xylazine and ketamine at the doses of 10 mg xylazine/kg b.w. and 100 mg ketamine/kg b.w.
- Animals fasted: Yes, prior to the clinical-chemical examinations, the animals were fasted for about 18 – 19 hours. Blood samples were taken from the heart as part of the procedure for euthanasia of the animals.
- How many animals: 5 adult males and 5 adult females from each group (0, 1, 2, 3, 0 SAT, and 3 SAT). Additionally 5 other males (from all groups) and 7 other females (from group 0, 1, 2, 3) or 5 females (from 0 SAT and 3 SAT group) were subjected to hematological (without bone marrow) biochemical and enzymatic examinations.. The level of thyroid hormone, i.e. total T4 (TT4) in serum was determined in all adult males after termination.
- Parameters examined: numbers of leukocytes, erythrocytes, and thrombocytes, the concentration of hemoglobin, hematocrit, MCV (mean corpuscular volume), MCH (mean corpuscular hemoglobin weight), and MCHC (mean corpuscular hemoglobin concentration), number of reticulocytes, circulatory blood and bone marrow microscopic examinations, Prothrombin time (PT) AND Partial thromboplastin time (APTT).
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at termination of the experiment, blood was collected for clinical-chemical examinations.
- Animals fasted: Yes, prior to the clinical-chemical examinations, the animals were fasted for about 18 – 19 hours.
- How many animals: 5 adult males and 5 adult females from each group (0, 1, 2, 3, 0 SAT, and 3 SAT). Additionally 5 other males (from all groups) and 7 other females (from group 0, 1, 2, 3) or 5 females (from 0 SAT and 3 SAT group) were subjected to hematological (without bone marrow) biochemical and enzymatic examinations.. The level of thyroid hormone, i.e. total T4 (TT4) in serum was determined in all adult males after termination.
- Parameters examined: - total protein, albumin, globulin (albumin subtracted from total protein, total cholesterol, urea nitrogen, creatinine, glucose, bilirubin, bile acids, sodium, potassium, chlorides and calcium. Also aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (AP).
URINALYSIS: Yes
- Time schedule for collection of urine: On the last day of the experiment
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined.: colour, volume (mL), specific gravity, pH, protein (g/L), glucose (mmol/L), ketone bodies (mmol/L), bilirubin (qualitative test), blood (Ery/μL), urobilinogen (μmol/L), leukocytes (leu/μL), urine sediment, squamous and columnar epithelium, leukocytes, erythrocytes, and crystals were determined.
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Males from group 0, 1, 2 and 3 were examined at the end of the administration period (1-2 days before euthanasia). Females from group 0, 1, 2 and 3 were examined during the last week of lactation (1 day before euthanasia). The behavioural studies on the males and females from satellite groups were performed at the end of the administration period (measurement 1) and then before the end of the additional observation period (measurement 2).
- Dose groups that were examined: 5 adult males and 5 adult females from each group were subjected to the studies.
- Battery of functions tested: observations of animal behaviour in an open field, responses to stimuli, locomotor activity, and forehand hindlimb grip strength.
IMMUNOLOGY: Yes
- Time schedule for examinations:
- How many animals: 5 adult males and 5 adult females
- Dose groups that were examined: from each group (0, 1, 2, 3, 0 SAT, and 3 SAT).
- Parameters examined.: The evaluation of the immune system was based on the results of clinical-chemical examinations and
post-mortem examinations (gross and histopathological examinations) of the spleen, thymus, and lymph nodes, as well as the statistical analysis of the absolute and relative weights of the spleen and thymus. The clinical-chemical parameters included blood morphology with a picture of peripheral blood and bone-marrow smears, the concentrations of albumin, total protein, the albumin/globulin ratio, creatinine, urea nitrogen, cholesterol, total bilirubin, and the activity of aspartate aminotransferase,
alanine aminotransferase, and alkaline phosphatase.
OTHER - HORMONE TESTING:
The level of thyroid hormone, i.e. total T4 (TT4) in serum were determined. The Mouse/Rat Thyroxine (T4) ELISA kit produced by Calbiotech were used.
Blood samples were collected from:
- all dams,
- all adult males after termination.
Blood samples from adult males were measured for TT4. There was no statistically significant changes in the hormone level, absolute/relative weight of the thyroid and histopathological changes of the thyroid connected with the test item, therefore further measurements of TT4 (dams) were not made - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes (see corresponding table)
A pathological examination involved macroscopic examinations of tissues and organs and a statistical analysis of absolute and relative weights of selected organs.
All adult animals (128 animals: 60 males and 68 females) used in the experiment survived the experiment and after being euthanized were examined pathologically.
The animals used in the study were subjected to the detailed gross examination which involved observations of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with all organs and their content.
Weights of organs of the animals used in the study were measured. Absolute and relative weights of brain, pituitary gland, thymus, thyroid gland, heart, liver, spleen, kidneys and adrenals from all adult rats from each group were recorded.
HISTOPATHOLOGY: Yes (see corresponding table)
The aim of the histopathological examination was to provide information on influence of the test item on tissues and organs of tested rats in following contexts:
determination of potential significant lesions in the brain (cerebrum, cerebellum, pons), spinal cord, eye with the optic nerve, pituitary gland, thyroid with parathyroids, stomach, small and large intestines including Peyer’s patches (duodenum, jejunum, ileum, cecum, colon), pancreas, liver, kidneys, ureters, adrenals, spleen, heart, aorta, thymus, trachea, lungs with the bronchi, urinary bladder, gonads: testes and ovaries with oviducts, uterus with the cervix, epididymides, accessory sex organs (prostate, seminal vesicles and coagulating glands), vagina, skin, mammary gland, lymph nodes, skeletal muscle with the peripheral nerve and femur with knee joint.
It has been conducted full histopathological examination of organs and tissues of adult animals of the control groups (group 0 and group 0 SAT) and the ones treated with the test item at the highest dose (group 3 and group 3 SAT). - Statistics:
- The results (average values and standard deviation) are presented in tables. The treated groups, i.e. groups 1, 2, and 3 were compared to the control group. The treated satellite group, i.e. 3 SAT was compared to group 0 SAT. Each sex was analyzed separately.
Clinical studies results: The clinical results were statistically analyzed using the one-way analysis of variance and the Dunnet’s test (group 0, 1, 2 and 3) and Student’s t-test (group 0 SAT, 3 SAT), (p ≤ 0.05).
Clinical-chemical examinations results: The results obtained in the groups 0, 1, 2 and 3 were statistically analysed using the one-way analysis of variance or Kruskal-Wallis test (in the absence of normal distribution or heterogeneous variances) and then Dunnet’s test (p ≤ 0.05). The results of the clinical-chemical examinations in satellite group were statistically analyzed using Student’s t test (p ≤ 0.05) or U Mann-Whitney test (in the absence of normal distribution). Some results of the urinalysis such as bilirubin and urine sediment were statistically analyzed using nonparametric statistics test - Kruskal-Wallis and then Dunnet’s test (p ≤ 0.05) (group 0, 1, 2, 3) and U Mann-Whitney test (group 0 SAT, 3 SAT), (p ≤ 0.05).
Post-mortem examinations results: The results, i.e. absolute and relative weights of internal organs as well as the numbers of implantation sites and corpora lutea were statistically evaluated using Dunnett’s test for groups 0, 1, 2, 3 and student’s T-test for satellite groups 0 SAT and 3 SAT (at p≤0.05).
Statistical analyses were conducted using the Microsoft Excel 2010 and the Statistica 10 programmes. - Clinical signs:
- no effects observed
- Description (incidence and severity):
- There were no differences in physical appearance and behaviour between the treated and the control groups. The animals didn’t show any clinical signs during the entire experiment. The only exception was the thinning of hair all over the body in one female from group 1 but this change is not considered to be treatment-related.
- Mortality:
- no mortality observed
- Description (incidence):
- No mortality and morbidity of the adult animals was observed.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no statistically significant differences in body weights between parental animals from treated and control groups.
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- There were no statistically significant differences in food intake by males and also by females from the treated (1, 2, 3, 3 SAT) and the control groups (0, 0 SAT).
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Description (incidence and severity):
- The ophthalmic examinations did not reveal any pathological changes.
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Hematological examinations: In case of peripheral blood examinations of animals from groups 1, 2 and 3, statistically significant changes in females of group 1 were stated (statistically significant: decrease in the hemoglobin concentration, decrease in the hematocrit value and decrease in the number of erythrocytes). Mentioned above changes were considered as accidental due to no confirmation in higher dosage i.e. group 2 and 3. There were no changes in males of group 1, 2, and 3. In case of peripheral blood examinations of males and females from group 3 SAT, statistically significant increase in the number of thrombocytes was stated compared to the control group (0 SAT). However, number of thrombocytes in group 3 SAT compared to the group 0 was at a similar level, so this change should not be connected with the test item administration. Statistically significant increase in the MCHC value in females of group 3 SAT was considered as accidental because this change was isolated, and no other deviations in red blood cells parameters were stated. In bone marrow examinations, statistically significant decrease in the number of proerythroblasts was stated in males of group 3 SAT. This change was regarded as random because there were no changes on the next stages of erythroblast development. Statistically significant decrease in the number of filamented eosinophils (bone marrow examination) in males of group 3SAT was also considered as accidental.
Coaglulological examinations: The treatment with the test item at three different dose levels did not affected coagulological parameters. - Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Biochemical examinations: In case of biochemical parameters, statistically significant increase in the chloride concentration in males of group 1, 2 and 3 were observed. Increase in chloride concentration was small, by about 2% compared to the control group and did not increase with increasing dose. For this reason statistically significant change which was observed may be considered as not treatment-related. These change was not observed in the satellite group, after 14-days post-treatment. In males of group 3 statistically significant increase in the calcium concentration was stated, it can be also considered as random due to low intensity. Statistically significant increase in the creatinine concentration was stated in females of group 3. This change was considered as accidental because the histopathological examination of the kidneys revealed no changes considered as treatment related. Statistically significant increase in the total protein concentration and decrease in the glucose concentration was stated in males of group 3 SAT compared to the control group (0 SAT) and those changes could be random. In case of females of group 3 SAT, statistically significant decrease in the urea nitrogen concentration and increase in the sodium concentration were stated and those changes were considered as accidental due to change in sodium was small (increase by about 0.9% compared to the group 0 SAT) and change in urea nitrogen is in the range of variability of the control group.
Enzymatic examinations: All changes observed in the males and females of group 1 and 2 (statistically significant decrease in the AP activity in males of group 1 and 2 and statistically significant decrease in the AST activity in females of group 2) were considered as random due to no confirmation in the higher dosage group. Statistically significant decrease in AST activity in males of group 3 SAT was consider as accidental, because in case of toxic impact on the liver function, an increase in activity of AST is observed. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Statistically significant decrease in the protein level was stated in females of group 1, 2 and 3 but this is not an adverse change so it will not be taken into account. Statistically significant increase in the level of urine protein in males of group 3SAT was stated and this change was considered as accidental.
Statistically significant decrease in the level of urine bacteria in males of group 3SAT is not an adverse change so it will not be taken into account. - Behaviour (functional findings):
- effects observed, non-treatment-related
- Description (incidence and severity):
- The behavioural studies showed that the nervous system functioned properly at dose levels of 33.3, 100, and 300 mg/kg b.w./day.
Open field observations: In case of males and females from groups 1, 2, 3 and control group, no involuntary clonic and tonic movements, changes in gait, or stereotypical behaviour were observed. A slight decrease in arousal was noticed in two males from group 0 (no. 4 and no. 5) and one female (no. 11) from group 3. A slight increase in arousal was noticed in one male (no. 3) from group 1, one male (no. 5) and three females (no. 12, no. 15 and no. 16) from group 2 and two females (no. 12 and no. 14) from group 3. A statistically significant increase of horizontal locomotor activity was noticed in males and females from group 2 compared to the control group.
A slight increase in arousal was noticed in one male (no. 34) and one female (no. 44) from group 0 SAT. A slight decrease in arousal was observed in one male (no. 34) and one female (no. 42) from group 0 SAT and in two males (no. 32 and no. 33) and two females (no. 43 and no. 45) from group 3 SAT. A moderate decrease in arousal was observed in two males (no. 31 and no. 32) and one female (no.44) from group 0 SAT and in three males (no. 31, no. 34 and no. 35) from group 3 SAT. A slight increase in arousal was observed in one female (no. 42) from group 3 SAT. There were statistically significant increase in the numbers of urine pools left by males from group 3 SAT compared to the group 0 SAT.
Evaluation of sensorimotor responses to stimuli: No differences between groups.
Measurement of the fore- and hindlimb grip strength: A statistically significant increase of the forelimb grip strength was observed in females from group 3
SAT compared to the control group (0 SAT). The hindlimb grip strength in females from group 3 SAT and 0 SAT was similar. - Immunological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No statistically significant changes considered as test item related were found in the results of clinicalchamical examinations therefore it can be stated that the test item did not affect the immune system. The gross and histopathological examination did not reveal any harmful effects of the test item on the immune system. Although cases of histopathological lesions in the immune system were observed, they should not be linked as the effect of the action of the test item since they were consider accidental and / or occurred in both, the treated and the control animals. There were also stated no incidence of overt infections, and occurrence of significant tumors in the treated rats. It was concluded that the test item is not immunotoxic for rats.
The observed changes are the following:
Statistically significant change was stated in the creatinine concentration in females of group 3. The macroscopic examination of the thymus, spleen, and lymph nodes did not reveal any pathological changes in any group with the exception of ecchymosis of thymus, petechiae on thymus and decreased of thymus. This changes occurred in a few animals both in control and treated groups. Hence, it was defined as accidental and should not be associated with the action of the test item.
Statistically significant differences, i.e. decrease in absolute weights of spleen in males from group 3 and decrease in relative weights of spleen in males from groups 1, 2 and 3. There were no other statistically significant changes in absolute and relative weights of thymus, spleen, and lymph nodes of males and females in all treated groups. - Organ weight findings including organ / body weight ratios:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The analysis of absolute and relative weights of internal organs of the males from groups 0, 1, 2, and 3 showed statistically significant decreases in absolute weights of pituitary gland and spleen in males from group 3 and in relative weights of pituitary gland and thyroid in males from group 3, as well as in relative weights of spleen in males from groups 1, 2 and 3.
The analysis of absolute weights of internal organs of the females from groups 0, 1, 2, and 3 showed statistically significant decreases in absolute weight of pituitary gland in females from group 2 and 3. There were no statistically significant changes in relative weights of organs of females in all treated groups.
There were no statistically significant changes in absolute and relative weights of organs in both satellite groups (0 SAT and 3 SAT) in males. Similarly there were no statistically significant changes in absolute weights of organs in both satellite groups (0SAT and 3SAT) in females. But there was statistically significant increase in relative weight of kidneys in females of group 3 SAT. - Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no significant lesions in the treated rats stated. The lesions (gross as well as histopathological) which were occurred in some organs and tissues, were mostly scattered among the treated animals and the control or occurred individually (all differences between treated and control groups were considered accidental, since they were of small magnitude, sporadic, not dose-related, and/or not consistent between sexes). On the basis of the results of the histopathological examination of organs and tissues it may be concluded that all of the revealed lesions were not associated with the test item.
The post-mortem examination revealed:
Group 0: Trachea filled with foamy liquid in 2 rats (females), Thymus petechiae in 1 rat (female), decreased in 1 rat (female), Lungs increased in 3 rats (2 males and 1 female), bright in 1 rat (female), bright foci above the surface in 3 rats (females), marbled in 5 rats (2 males and 3 females), dark red in 1 rat (male), dark red foci in 3 rats (1 male and 2 females), dark red area in 7 rats (1 male and 6 females), Heart slightly increased in 1 rat (male), Uterus congestion in 1 rat(female).
Group 1: Trachea filled with foamy liquid in 2 rats (females), Thymus ecchymosis of 1/3 organ in 1 rat( female), petechiae in 1 rat (female), decreased in 2 rat (females), Lungs increased in 8 rats (3 males and 5 females), bright in 1 rat (female), bright foci above the surface in 5 rats (females), foci collapsed under the surface in 1 rat (female), red in 1 rat (female), marbled in 2 rats (females), dark red in 3 rats (males), dark red foci in 1 rat (female), dark red area in 3 rats (1 male and 2 females), Heart slightly increased in 1 rat (female), small intestine congestion in 1 rat (female), Cecum congestion in 1 rat (female), Uterus congestion in 1 rat (female), cyanosis of the horns in 1 rat (female).
Group 2: Trachea filled with foamy liquid in 1 rat (female), Thymus petechiae in 1 rat (female) decreased in 1 rat (female), Lungs increased in 2 rats (females), bright in 1 rat (female), bright foci above the surface in 1 rat (female), marbled in 5 rats (females), dark red foci in 1 rat (female), dark red area in 4 rats (2 males and 2 females), Heart slightly increased in 1 rat (female), Ovaries congestion in 1 rat (female), Uterus congestion in 2 rats (females), partial cyanosis in 1 rat (female), expanded in 1 rat (female), cyanosis of the horns in 3 rats (female).
Group 3: Thymus petechiae in 1 rat (female), Lungs increased in 3 rats (2 males and 1 female), bright in 1 rat (female), bright foci above the surface in 1 rat (female), marbled in 5 rats (1 male and 4 females), dark red in 1 rat (female), dark red area in 4 rats (female), Uterus congestion in 1 rat (female), expanded in 1 rat (female), thickened wall in 1 rat (female), filled with clear liquid in 1 rat (female).
Group 0 SAT: Trachea filled with foamy liquid in 2 rats (1 male and 1 female), Thymus petechiae in 8 rats (3 males and 5 females), decreased in 2 rats (1 male and 1 female), Lungs bright red in 5 rats (4 males and 1 female), bright red foci in 2 rats (1 male and 1 female), bright red foci above the surface in 4 rats (2 males and 2 females), increased in 3 rats (2 males and 1 female), marbled in 7 rats (6 males and 1 female), dark red foci in 9 rats (3 males and 6 females), Heart increased in 4 rats (males), Liver lesion 7mm long, yellowish, grainy on the visceral side of the left lobe in 1 rat (female), Spleen increased in 1 rat (male)
Cecum, congestion in 1 rat (male), Colon petechiae in 1 rat (male), Kidneys increased in 2 rats (males), Ovaries congestion in 1 rat (female), Uterus congestion in 1 rat (female), slightly expanded in 4 rats (females), filled with clear liquid in 4 rats (females).
Group 3 SAT:
Trachea filled with foamy liquid in 4 rats (3 males and 1 female), Thymus petechiae in 9 rats (5 males and 4 females), decreased in 1 rat (male), Lungs bright red in 3 rats (males), bright red foci above the surface in 5 rats (3 males and 2 females), increased in 3 rats (2 males and 1 female), marbled in 8 rats (7 males and 1 female), dark red in 1 rat (male), dark red foci in 9 rats (1 male and 8 females), dark red foci under the surface in 2 rats (males), doughy consistency in 1 rat (female),Heart increased in 2 rats (males), congestion in 2 rats (males),mandibular lymph nodes petechiae in 3 rats (1 male and 2 females), Ovaries congestion in 1 rat (female), Uterus congestion in 1 rat (female), slightly expanded in 2 rats (females), filled with clear liquid in 2 rats (females), Other lesions yellowish, elastic, size 5x2mm in subcutaneous tissue of cervical area above the salivary gland in 1 rat (male), yellowish, 5mm long located in fat tissue in left ovary area in 1 rat (female), yellowish 5mm long located in fat tissue in pelvis cavity inlet area in 1 rat (female), Cranial cavity filled with blood in 1 rat (male) - Neuropathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no significant histopathological lesions observed in the animals treated with the highest dose of the test item (groups: 3 and 3 SAT) therefore the full histopathological examination was not extended to organs and tissues of the animals exposed to the mid and the low dose of the test item. As for the animals of the satellite group (group 3 SAT) there were no delayed significant changes in organs and tissues for 14-days additional observation after the treatment.
Changes observed were:
Group 0 - pituitary: microcyst in 2 males, thyroid: parathyroid hypertrophy in 1 female, thymus: congestion in 1 male, effusion in 1 female, lymphocytic tissue depletion in 1 female, heart: erythrocytorrhagia in 3 males, leukocyte infiltration in 1 male, foci of fibrosis in 1 male, foci of fatty degeneration in 1 male, left and right ventricular myocardium hypertrophy in 1 male, lungs: congestion in 1 male, erythrocytorrhagia in 5 males and 9 females, edema in 3 males and 4 females, emphysema in 4 males and 9 females, foamy cell infiltration in 2 males and 7 females, lymphocyte infiltration in 1 male, leukocyte infiltration in 1 male, liver: congestion in 1 male and 2 females, - effusion in 1 male, bile ducts proliferation in 1 male, Browicz-Kupffer cell proliferation in 1 male, hepatocyte fine-droplet fatty degeneration in 1 male, spleen: congestion in 1 female, depletion of the white pulp in 2 females, pancreas: islets of Langerhans hypertrophy in 1 male and 1 female, ducts proliferation in 1 female, jejunum: lymphatic follicle reactive hyperplasia in 1 male, ileum: lymphocyte infiltration in 1 female, cecum: lymphatic follicle reactive hyperplasia in 1 male, colon: lymphatic follicle reactive hyperplasia in 1 male, kidneys: erythrocytorrhagia in 1 male and 1 female, glomeruli congestion in 6 males and 4 females, lymphocyte infiltration in 2 females, glomeruli hypertrophy in 9 males and 7 females, glomeruli atrophy in 4 males and 5 females, Bowman's capsule extension in 1 male and 2 females, retention microcysts in 2 females, hyaline casts in 1 female, prostate: congestion in 2 males, edema in 2 males, lymphocyte infiltration in 5 males, mandibular lymph nodes: congestion in 1 male, hyperplasia in 1 male, lymphocytic tissue depletion in 1 female, mesenteric lymph nodes: hyperplasia in 1 male, lymphocytic tissue depletion in 1 female.
Group 1 - thymus: congestion in 1 female, effusion in 1 female, heart: erythrocytorrhagia in 1 female, left ventricular myocardium hypertrophy in 1 female, lungs: congestion in 3 males and 2 females, erythrocytorrhagia in 4 males and 10 females, edema in 3 males and 2 females, emphysema in 10 females, foamy cell infiltration in 3 females, prostate: lymphocyte infiltration in 3 males, uterus: congestion in 1 female, effusion in 2 females, fibroid in 2 females.
Group 2: thymus: lymphocytic tissue depletion in 1 female, heart: left ventricular myocardium hypertrophy in 1 female, lungs: congestion in 1 male and 2 females, erythrocytorrhagia in 3 males and 9 females, edema in 2 females, emphysema in 1 male and 3 females, foamy cell infiltration in 4 females, prostate: lymphocyte infiltration in 6 males, uterus: congestion in 2 females, effusion in 2 females, fibroid in 1 female.
Group 3: thyroid: congestion in 1 male, thymus: congestion in 2 males, erythrocytorrhagia in 1 male, heart: erythrocytorrhagia in 1 male, leukocyte infiltration in 1 male, lungs: congestion in 2 males and 2 females, erythrocytorrhagia in 6 males and 8 females, edema in 2 males and 1 female, emphysema in 5 males and 3 females, foamy cell infiltration in 1 male and 1 female, lymphocyte infiltration in 1 male, liver: congestion in 1 male, pancreas: islets of Langerhans hypertrophy in 3 males, cecum: lymphatic follicle reactive hyperplasia in 1 female, colon: lymphatic follicle reactive hyperplasia in 2 males and 1 female, kidneys: erythrocytorrhagia in 1 male, glomeruli congestion in 10 males and 6 females, lymphocyte infiltration in 1 male, glomeruli hypertrophy in 9 males and 9 females, glomeruli atrophy in 10 males and 8 females, Bowman's capsule extension in 6 males, retention microcysts in 1 male and 2 females, adrenals: left adrenal gland atrophy with medulla microcyst; right adrenal gland hyperplasia in 1 female, urinary bladder: lymphocyte infiltration in 5 males, prostate: congestion in 2 males, lymphocyte infiltration in 6 males, mesenteric lymph nodes: congestion in 1 male, hyperplasia in 1 male.
Group 0 SAT - pituitary: hypertrophy in 1 female, microcyst in 2 males, thymus: congestion in 2 males, effusion in 1 male and 3 females, heart: congestion in 1 male, erythrocytorrhagia in 1 male, foci of fibrosis in 2 males, myocardium of the left ventricular hypertrophy in 3 males, lungs: congestion in 4 males, erythrocytorrhagia in 8 male and 9 females, edema in 3 males, emphysema in 5 male and 4 females, foamy cell infiltration in 1 female, lymphocyte infiltration in 1 male, inflammatory infiltration in 1 male, foci of fibrosis in 1 male, liver: congestion in 1 male, Browicz-Kupffer cell proliferation in 1 male and 1 female, focus of fibrosis in 1 female, spleen: congestion in 1 male, pancreas: islets of Langerhans congestion in 2 males, islets of Langerhans hypertrophy in 7 males and 2 females
- lymphocyte infiltration in 2 males and 1 female, jejunum: lymphatic follicle reactive hyperplasia in 1 female, cecum: lymphatic follicle reactive hyperplasia in 1 male, colon: lymphatic follicle reactive hyperplasia in 1 female, lymphocyte infiltration in 1 female, kidneys: glomeruli congestion in 5 males and 4 females, glomeruli hypertrophy in 6 males and 4 females, glomeruli atrophy in 3 males and 2 females, Bowman's capsule extension in 1 male, retention microcysts in 1 male and 2 females, prostate: congestion in 1 male, lymphocyte infiltration in 3 males, ovaries: congestion in 1 female, mandibular lymph nodes: lymphocytic tissue reactive hyperplasia in 1 female.
Group 3 SAT - thymus: congestion in 2 males and 1 female, effusion in 2 males and 4 females, heart: congestion in 3 males, erythrocytorrhagia in 4 males and 1 female, foci of fibrosis in 3 males, myocardium of the left ventricular hypertrophy in 2 males, lungs: congestion in 4 males and 1 female, erythrocytorrhagia in 9 males and 9 females, edema in 4 males and 1 female, emphysema in 4 males and 4 females, foamy cell infiltration in 1 male and 1 female, liver: congestion in 1 male, effusion in 1 male, spleen: congestion in 1 male, hemosiderin deposits in 1 male and 2 females, pancreas: islets of Langerhans congestion in 2 males and 2 females, islets of Langerhans hypertrophy in 3 males and 3 females, foci of atrophy of lobules with fibrosis in 1 male, ducts proliferation in 1 male, cecum: lymphatic follicle reactive hyperplasia in 2 males and 3 females, colon: lymphocyte infiltration in 1 female, kidneys: glomeruli congestion in 5 males and 6 females, glomeruli hypertrophy in 6 males and 5 females, glomeruli atrophy in 3 males and 3 females, Bowman's capsule extension in 1 male, urinary bladder: lymphocyte infiltration in 1 male, prostate: lymphocyte infiltration in 2 males, mandibular lymph nodes: congestion in 1 male and 1 female, erythrocytorrhagia in 1 female, effusion in 1 male and 1 female, gross lesion isolated from subcutaneous tissue of cervical area above the salivary gland (nodule): lipoma in 1 male gross lesion isolated from the fat tissue of the left ovary area (nodule): lipoma in 1 female, gross lesion isolated from the fat tissue of the pelvis cavity inlet area (nodule): lipoma in 1 female. - Histopathological findings: neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were also observed individual cases of small benign tumors such as lipomas of the adipose tissue and fibroids of the uterus. In the subcutaneous tissue of region of cervical area above the salivary gland (male in the group 3 SAT) and region of the abdominopelvic cavity (two females in the group 3 SAT) the small lipomas were occurred. As for the uterus (two females in the group 1 and one female in the group 2) the fibroids were observed. The occurrence of them was clearly incidental to the treatment and should not be linked with the test item.
Both, the lipomas and fibroids were considered as spontaneous lesions, consistent with background findings commonly observed in rats. The occurrence of the left adrenal gland atrophy with medulla microcyst and right adrenal gland hyperplasia in one of the females in the group 3 is also accidental. It seems to be a kind of a malformation. - Other effects:
- no effects observed
- Description (incidence and severity):
- Hormonal examinations: Statistically significant changes in the TT4 level, absolute/relative weight of the thyroid and histopathological changes of the thyroid related with the test item in all males were not observed, so the determination of TT4 levels were not extended to females.
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 300 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No test item related adverse effects observed at the highest dose tested.
- Key result
- Critical effects observed:
- no
- Conclusions:
- The oral 28d-NOAEL in rats was determined to be 300 mg/kg bw/day since no adverse effects were observed at the highest dose tested.
- Executive summary:
A Combined repeated dose toxicity study with the reproduction/developmental toxicity screening test was conducted in rats according to the OECD Guideline No. 422 (2016) with GLPs. The study involved the determination of the effects of the test item on body weights, food intake, behaviour, clinical, haematological, coagulation, biochemical, enzymatic, urine, and hormonal parameters and the evaluation of gross and histopathological lesions in tissues and internal organs. Two dose range finding tests and the main study were conducted. The first Dose range finding stated that 7-day exposure of rats to VL3 at the doses of 250, 500 and 1000 mg/kg b.w./day cause one treatment-related change, i.e. decrease in number of leukocytes (males from group 1 and females from group 1 and 3).The second Dose-range finding test revealed that 7-day exposure of rats to VL3 at the doses of 31.3, 62.5 and 125.0 mg/kg b.w./day did not cause treatment-related changes. The main study was performed on 60 males and 68 females of Wistar rats treated with vehicle control or with 33.3, 100, 300 mg/kg b.w./day of test item. The animals received the test item/vehicle control by gavage once a day (7 days/week). After two weeks of initial treatment, the animals were mated. The test item/vehicle control was administered to males for 28 days (2 weeks prior to mating, during the mating and post-mating period). Females were dosed throughout the study. This included 2 weeks prior to mating, the variable time to conception, the duration of pregnancy, and at least 13 days after delivery (about 51-59 days, depending on the duration of the mating and pregnancy period). Furthermore, two satellite groups were used;: a control group and one treated group which was given VL3 at the dose of 300 mg/kg b.w/day. The satellite groups were not be mated. Males from groups 0 SAT and 3 SAT were treated for 28 days. Females from groups 0 SAT and 3 SAT were treated for 53 days (on average as long as females from groups 0, 1, 2 and 3). After that, males and females from satellite groups were observed for 14 days to evaluate the reversibility, stability, or delay in the onset of possible harmful effects. During the experiment, no mortality of the adults was observed. There were no differences in physical appearance and behaviour of adult animals between the treated and the control groups. No treatment-related clinical signs, ophthalmic alterations or significant differences in body weights and food intake were observed. Statistical analysis of the clinical-chemical examinations results obtained in males and females of group 1, 2 and 3 and satellite groups revealed some statistically significant changes however all of them were considered as not related with the administration of the test item. There were gross changes observed in control group 0, treated groups 1, 2 and 3 and satellite groups 0 SAT and 3 SAT. However, these changes occurred regardless of the degree of exposure to the test item (this changes were observed both in control and treated groups). Statistical analysis of absolute and relative weights of internal organs revealed statistically significant changes but these organ weight disorders seem to be accidental. The macroscopic and the histological examinations of the rats treated with the test item at the doses of 33.3, 100, and 300 mg/kg b.w./day did not reveal any toxic effects. Additionally, based on results, it was concluded that the test item is not immunotoxic for rats.
In conclusion, the treatment of adult (male and female) rats with the test item at the dose levels of 33.3, 100 and 300 mg/kg bw/day by the repeated oral administration, revealed no adverse effects. Based on the results of clinical studies, clinical-chemical, pathomorphological and histopathological examinations, the no observed adverse effect level (NOAEL) of the test item for systemic toxicity was determined to be 300 mg/kg b.w./day.
Referenceopen allclose all
Table 4. Mortality and clinical observations – main groups
Parameter | GROUP / DOSE [mg/kg b.w./day] |
| ||||||
0 / 0 | 1 / 62.5 | 2 / 250 | 3 / 1000 | |||||
males | females | males | females | males | females | males | females | |
Animals in group | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Death of animals | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 |
Euthanized animals | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 | 0 / 10 |
Animals with clinical signs | 2 / 10 | 1 / 10 | 1 / 10 | 1 / 10 | 0 / 10 | 0 / 10 | 3 / 10 | 5 / 10 |
CLINICAL SIGNS |
| |||||||
Survived animals |
| |||||||
Thinning coat or alopecia of particular part of body | 2 / 10 | 1 / 10 | 1 / 10 | 1 / 10 | 0 / 10 | 0 / 10 | 3 / 10 | 5 / 10 |
Table 5. Mortality and clinical observations – satellite groups
Parameter |
| GROUP / DOSE [mg/kg b.w./day] | |||
| 0 SAT / 0 | 3 SAT / 1000 | |||
males |
| females | males | females | |
Animals in group | 10 |
| 10 | 10 | 10 |
Death of animals | 0 / 10 |
| 0 / 10 | 0 / 10 | 0 / 10 |
Euthanized animals | 0 / 10 |
| 0 / 10 | 0 / 10 | 1 / 10 |
Animals with clinical signs | 0 / 10 |
| 1 / 10 | 2 / 10 | 2 / 10 |
CLINICAL SIGNS |
| ||||
Survived animals |
| ||||
Thinning coat or alopecia of particular part of body | 0 / 10 |
| 1 / 10 | 2 / 10 | 1 / 10 |
Euthanized animals |
| ||||
dejection and distinct decrease of locomotor activity, animal lied on its abdomen motionless, no reaction to external stimuli and catching, no intake of food and water, pallor of the skin and irises, bristled coat and hypothermia | 0 / 10 |
| 0 / 10 | 0 / 10 | 1 / 10 |
Table 6. Average food intake [g/100 g b.w./day] – main groups
Week | GROUP / DOSE [mg/kg b.w./day] |
| ||||||
0 / 0 | 1 / 62.5 | 2 / 250 | 3 / 1000 | |||||
♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | |
1 | 9.25 ± 0.21 | 8.60 ± 0.15 | 9.14 ± 0.27 | 8.69 ± 0.23 | 9.21 ± 0.28 | 8.60 ± 0.43 | 8.98 ± 0.23 | 8.75 ± 0.53 |
2 | 8.11 ± 0.09 | 8.11 ± 0.20 | 8.02 ± 0.20 | 8.37 ± 0.18 | 8.34 ± 0.53 | 7.92 ± 0.29 | 7.97 ± 0.34 | 8.04 ± 0.39 |
3 | 7.18 ± 0.37 | 7.61 ± 0.49 | 7.00 ± 0.33 | 7.84 ± 0.30 | 7.34 ± 0.31 | 7.32 ± 0.26 | 7.18 ± 0.25 | 7.49 ± 0.19 |
4 | 6.26 ± 0.36 | 6.90 ± 0.24 | 6.31 ± 0.28 | 7.34 ± 0.38* | 6.55 ± 0.20 | 6.76 ± 0.33 | 6.44 ± 0.25 | 6.95 ± 0.40 |
5 | 5.75 ± 0.31 | 6.38 ± 0.33 | 5.83 ± 0.30 | 6.81 ± 0.59 | 5.85 ± 0.09 | 6.19 ± 0.46 | 5.91 ± 0.26 | 6.48 ± 0.28 |
6 | 5.50 ± 0.09 | 6.28 ± 0.30 | 5.55 ± 0.22 | 6.50 ± 0.24 | 5.41 ± 0.08 | 6.08 ± 0.37 | 5.44 ± 0.25 | 6.23 ± 0.26 |
7 | 5.30 ± 0.22 | 5.81 ± 0.20 | 5.29 ± 0.22 | 6.18 ± 0.37 | 5.27 ± 0.09 | 5.78 ± 0.37 | 5.29 ± 0.26 | 5.79 ± 0.16 |
8 | 4.90 ± 0.32 | 5.46 ± 0.20 | 4.99 ± 0.22 | 5.75 ± 0.45 | 4.96 ± 0.14 | 5.39 ± 0.34 | 4.98 ± 0.32 | 5.41 ± 0.13 |
9 | 4.76 ± 0.22 | 5.24 ± 0.11 | 4.80 ± 0.27 | 5.46 ± 0.23 | 4.58 ± 0.14 | 5.31 ± 0.39 | 4.66 ± 0.28 | 5.33 ± 0.09 |
10 | 4.63 ± 0.22 | 5.26 ± 0.15 | 4.59 ± 0.17 | 5.49 ± 0.17 | 4.38 ± 0.14* | 5.21 ± 0.45 | 4.52 ± 0.14 | 5.31 ± 0.14 |
11 | 4.43 ± 0.17 | 5.08 ± 0.31 | 4.47 ± 0.07 | 5.32 ± 0.37 | 4.30 ± 0.12 | 4.99 ± 0.36 | 4.46 ± 0.18 | 5.09 ± 0.07 |
12 | 4.16 ± 0.27 | 5.17 ± 0.13 | 4.39 ± 0.11* | 5.20 ± 0.25 | 4.17 ± 0.18 | 4.96 ± 0.41 | 4.44 ± 0.24* | 5.10 ± 0.40 |
13 | 4.48 ± 0.18 | 5.38 ± 0.50 | 4.56 ± 0.03 | 5.41 ± 0.56 | 4.29 ± 0.24 | 5.32 ± 0.46 | 4.63 ± 0.30 | 5.07 ± 0.13 |
x ± SD
n - number of animals per group
* - statistically significant difference with p ≤ 0.05; Dunnett’s test;
Table 7. Average food intake [g/100 g b.w./day] – satellite groups
Week | GROUP / DOSE [mg/kg b.w./day] | |||
0 SAT / 0 | 3 SAT / 1000 | |||
♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | |
1 | 9.60 ± 0.05 | 9.01 ± 0.22 | 9.41 ± 0.28 | 8.66 ± 0.32** |
2 | 8.64 ± 0.18 | 8.30 ± 0.11 | 8.31 ± 0.26* | 8.27 ± 0.36 |
3 | 7.47 ± 0.13 | 7.79 ± 0.18 | 7.62 ± 0.24 | 7.95 ± 0.15 |
4 | 6.88 ± 0.20 | 7.16 ± 0.25 | 6.75 ± 0.07 | 7.17 ± 0.19 |
5 | 6.21 ± 0.25 | 6.55 ± 0.29 | 6.15 ± 0.11 | 6.53 ± 0.28 |
6 | 5.54 ± 0.13 | 6.28 ± 0.43 | 5.72 ± 0.06** | 6.44 ± 0.22 |
7 | 5.33 ± 0.03 | 5.90 ± 0.46 | 5.34 ± 0.18 | 6.19 ± 0.13 |
8 | 5.06 ± 0.05 | 5.56 ± 0.37 | 5.13 ± 0.08** | 5.68 ± 0.19 |
9 | 4.77 ± 0.15 | 5.23 ± 0.25 | 4.84 ± 0.05 | 5.46 ± 0.29 |
10 | 4.54 ± 0.13 | 5.35 ± 0.35 | 4.51 ± 0.11 | 5.45 ± 0.06 |
11 | 4.45 ± 0.20 | 5.33 ± 0.32 | 4.58 ± 0.08 | 5.39 ± 0.32 |
12 | 4.31 ± 0.04 | 5.00 ± 0.27 | 4.49 ± 0.11** | 5.01 ± 0.19 |
13 | 4.34 ± 0.18 | 4.92 ± 0.10 | 4.56 ± 0.11** | 5.01 ± 0.25 |
14 | 4.90 ± 0.05 | 5.60 ± 0.24 | 5.32 ± 0.22** | 5.81 ± 0.13 |
15 | 4.84 ± 0.26 | 5.66 ± 0.22 | 5.22 ± 0.18** | 5.84 ± 0.10** |
16 | 5.22 ± 0.17 | 5.85 ± 0.21 | 5.36 ± 0.21 | 6.02 ± 0.20# |
x ± SD
n - number of animals per group
* - statistically significant difference with p ≤ 0.05; Student’s t-test;
** - statistically significant difference with p ≤ 0.05; Mann-Whitney U test; # - n=7
Table 8. Urine sediment examinations - main groups
Parameter |
| GROUP / DOSE [mg/kg b.w.] |
| |||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |
| males | females |
| |||||
n=9# | n=10 | n=10 | n=10 | n=8# | n=9# | n=9# | n=9# | |
SQUAMOUS EPITHELIA | 1.33 ± 0.50 | 1.40 ± 0.52 | 1.30 ± 0.48 | 1.40 ± 0.52 | 1.00 ± 0.00 | 1.22 ± 0.44 | 1.22 ± 0.44 | 1.44 ± 0.53 |
TRANSITIONAL EPITHELIA | 0.00 ± 0.00 | 0.10 ± 0.32 | 0.00 ± 0.00 | 1.00 ± 0.82* | 0.00 ± 0.00 | 0.22 ± 0.44 | 0.11 ± 0.33 | 0.78 ± 0.44* |
LEUKOCYTES | 1.00 ± 0.00 | 1.00 ± 0.00 | 1.00 ± 0.00 | 2.50 ± 0.71* | 1.00 ± 0.00 | 1.00 ± 0.00 | 1.00 ± 0.00 | 2.00 ± 0.50* |
ERYTHROCYTES | 1.33 ± 0.50 | 1.30 ± 0.48 | 1.40 ± 0.52 | 1.40 ± 0.52 | 1.13 ± 0.35 | 1.22 ± 0.44 | 1.33 ± 0.50 | 1.56 ± 0.53 |
BACTERIA | 3.11 ± 0.78 | 2.90 ± 0.74 | 3.00 ± 0.82 | 3.50 ± 0.71 | 3.13 ± 1.13 | 2.78 ± 1.09 | 2.78 ± 0.44 | 2.67 ± 0.71 |
AMORPHOUS URATES | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
AMORPHOUS PHOSPHATES | 0.00 ± 0.00 | 0.00 ± 0.00 | 1.20 ± 1.99 | 1.10 ± 1.79 | 0.38 ± 1.06 | 1.22 ± 1.86 | 0.33 ± 1.00 | 0.00 ± 0.00 |
AM. MAG. PHOSPHATES | 2.11 ± 1.45 | 2.70 ± 0.82 | 2.50 ± 1.35 | 0.90 ± 1.20 | 1.13 ± 1.13 | 1.67 ± 0.87 | 1.89 ± 1.27 | 0.56 ± 1.13 |
Table 9. Urine sediment examinations - satellite groups
Parameter |
| GROUP / DOSE [mg/kg b.w.] | |||
0SAT/0 | 3SAT/1000 | 0SAT/0 | 3SAT/1000 | ||
| males | females | |||
n=10 | n=10 | n=10 | n=9 | ||
SQUAMOUS EPITHELIA | 1.10 ± 0.32 | 1.20 ± 0.42 | 1.10 ± 0.32 | 1.33 ± 0.50 | |
TRANSITIONAL EPITHELIA | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.22 ± 0.44 | |
LEUKOCYTES | 1.00 ± 0.00 | 1.10 ± 0.32 | 1.00 ± 0.00 | 1.00 ± 0.00 | |
ERYTHROCYTES | 1.20 ± 0.42 | 1.00 ± 0.00 | 1.10 ± 0.32 | 1.11 ± 0.33 | |
BACTERIA | 3.90 ± 0.74 | 3.20 ± 0.92 | 3.30 ± 1.06 | 3.00 ± 1.22 | |
AMORPHOUS URATES | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | |
AMORPHOUS PHOSPHATES | 1.30 ± 1.70 | 1.00 ± 1.63 | 1.30 ± 2.11 | 0.67 ± 1.32 | |
AM. MAG. PHOSPHATES | 2.30 ± 0.82 | 1.50 ± 1.35 | 2.10 ± 0.57 | 0.78 ± 1.20** | |
** - statistically significant difference with p ≤ 0.05; Mann-Whitney U test
Table 10. Locomotor activity – satellite groups
Parameter |
| GROUP / DOSE [mg/kg b.w./day] |
| |||||
0 SAT / 0 (measurement 1) | 3 SAT / 1000 (measurement 1) | 0 SAT / 0 (measurement 2) | 3 SAT / 1000 (measurement 2) | |||||
♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | ♂ n=10 | ♀ n=10 | |
DISTANCE 0- 30MIN [m] | 555.48 ± 120.95 | 630.76 ± 128.61 | 580.48 ± 130.47 | 670.88 ± 151.01 | 610.24 ± 93.32 | 626.40 ± 115.83 | 480.58 ± 55.26 * | 602.03 ± 142.87 |
DISTANCE 0- 10MIN [m] | 304.65 ± 35.52 | 329.55 ± 59.65 | 314.60 ± 71.50 | 328.80 ± 40.00 | 289.03 ± 59.91 | 304.18 ± 57.29 | 236.53 ± 33.50 * | 291.60 ± 48.12 |
DISTANCE 10-20MIN [m] | 169.04 ± 62.26 | 192.42 ± 54.80 | 165.10 ± 55.19 | 187.68 ± 56.46 | 182.28 ± 46.59 | 170.03 ± 47.26 | 164.18 ± 42.17 | 178.57 ± 72.34 |
DISTANCE 20-30MIN [m] | 81.77 ± 51.37 | 108.80 ± 45.60 | 100.84 ± 68.51 | 154.53 ± 74.05 | 138.93 ± 39.09 | 152.22 ± 55.01 | 79.87 ± 48.19* | 131.86 ± 42.35 |
NUMBER OF REARINGS 0-30MIN | 63.60 ± 18.88 | 86.50 ± 31.81 | 71.90 ± 14.72 | 105.20 ± 30.11 | 71.40 ± 19.67 | 88.30 ± 17.53 | 42.80 ± 12.86* | 102.10 ± 47.51 |
NUMBER OF REARINGS 0-10MIN | 40.40 ± 6.96 | 49.40 ± 17.28 | 49.00 ± 13.09 | 56.60 ± 13.95 | 43.80 ± 15.28 | 46.80 ± 13.85 | 26.60 ± 7.44 * | 51.70 ± 22.03 |
NUMBER OF REARINGS 10-20MIN | 17.10 ± 11.79 | 22.40 ± 8.33 | 14.10 ± 8.28 | 26.80 ± 13.11 | 17.00 ± 7.72 | 19.70 ± 7.09 | 10.50 ± 9.06 | 32.50 ± 21.88 |
NUMBER OF REARINGS 20-30MIN | 6.30 ± 6.88 | 14.90 ± 11.61 | 8.90 ± 9.62 | 21.80 ± 16.19 | 10.90 ± 4.56 | 21.80 ± 11.97 | 5.70 ± 7.90 | 17.90 ± 11.23 |
* – statistically significant difference at p ≤ 0.05, Student's t-test
Table 11. Hematological and coagulological examinations - main groups
Parameter | GROUP/DOSE [mg/kg b.w.] |
| ||||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |
males | females |
| ||||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=8# | n=10 | |
HEMOGLOBIN [g/L] | 153.10 ± 4.48 | 153.80 ± 3.65 | 154.90 ± 3.73 | 149.80 ± 3.91 | 146.60 ± 4.12 | 148.10 ± 5.53 | 147.00 ± 4.81 | 143.80 ± 8.57 |
HEMATOCRIT [1/1] | 0.429 ± 0.017 | 0.428 ± 0.010 | 0.431 ± 0.013 | 0.418 ± 0.012 | 0.418 ± 0.012 | 0.414 ± 0.020 | 0.412 ± 0.016 | 0.411 ± 0.027 |
ERYTHROCYTES [x 1012/L] | 8.91 ± 0.42 | 8.96 ± 0.18 | 9.09 ± 0.30 | 9.20 ± 0.33 | 8.04 ± 0.25 | 8.17 ± 0.32 | 8.11 ± 0.16 | 8.06 ± 0.55 |
MCV [fL] | 48.12 ± 0.99 | 47.75 ± 0.56 | 47.41 ± 1.10 | 45.50 ± 1.27* | 51.95 ± 0.74 | 50.70 ± 1.36 | 50.83 ± 1.79 | 51.04 ± 1.26 |
MCH [pg] | 17.20 ± 0.38 | 17.15 ± 0.24 | 17.04 ± 0.40 | 16.30 ± 0.39* | 18.26 ± 0.27 | 18.16 ± 0.38 | 18.11 ± 0.51 | 17.84 ± 0.46 |
MCHC [g/L] | 357.50 ± 4.74 | 359.40 ± 3.47 | 359.90 ± 3.57 | 358.40 ± 3.17 | 351.70 ± 2.75 | 358.20 ± 5.73 | 356.75 ± 8.66 | 349.90 ± 8.50 |
LEUKOCYTES [x 109/L] | 4.10 ± 0.51 | 3.59 ± 0.96 | 4.38 ± 0.59 | 4.05 ± 0.93 | 2.92 ± 0.66 | 2.65 ± 0.45 | 3.11 ± 0.76 | 2.88 ± 0.58 |
THROMBOCYTES [x 109/L] | 507.10 ± 55.42 | 488.90 ± 42.66 | 513.70 ± 38.15 | 508.00 ± 30.01 | 485.80 ± 37.28 | 446.80 ± 110.86 | 482.75 ± 33.96 | 453.30 ± 90.89 |
RETICULOCYTES [1/1] | 0.010 ± 0.001 | 0.009 ± 0.002 | 0.008 ± 0.002 | 0.008 ± 0.001* | 0.011 ± 0.003 | 0.011 ± 0.002 | 0.011 ± 0.003 | 0.009 ± 0.005 |
PT [s] | 15.99 ± 0.86 | 14.48 ± 3.13 | 15.06 ± 1.14 | 14.91 ± 1.38 | n=9# | 13.11 ± 1.00 | 13.36 ± 1.37 | n=8# |
12.99 ± 1.07 | 13.85 ± 1.69 | |||||||
APTT [s] | 24.09 ± 8.94 | 26.02 ± 5.58 | 29.35 ± 17.12 | 25.10 ± 4.25 | n=9# | 19.77 ± 3.70 | 22.58 ± 5.61 | n=8# |
20.70 ± 5.26 | 21.19 ± 5.62 | |||||||
x ± SD; n - number of test animals;
* - statistically significant difference with p ≤ 0.05; Dunnett's test;
# - n<10 because of clot in the sample
Table 12. Haematological and coagulological examinations - satellite groups
Parameter | GROUP/DOSE [mg/kg b.w.] | |||
0SAT/0 | 3SAT/1000 | 0SAT/0 | 3SAT/1000 | |
males | females | |||
n=10 | n=9# | n=10 | n=9 | |
HEMOGLOBIN [g/L] | 154.80 ± 2.90 | 154.33 ± 5.74 | 151.40 ± 3.47 | 149.33 ± 2.06 |
HEMATOCRIT [1/1] | 0.438 ± 0.014 | 0.441± 0.016 | 0.431 ± 0.014 | 0.433 ± 0.005 |
ERYTHROCYTES [x 1012/L] | 8.98 ± 0.26 | 9.07 ± 0.36 | 8.41 ± 0.23 | 8.16 ± 0.13* |
MCV [fL] | 48.71 ± 1.20 | 48.58 ± 0.67 | 51.27 ± 0.95 | 53.11 ± 0.86* |
MCH [pg] | 17.25 ± 0.32 | 17.03 ± 0.24 | 18.01 ± 0.29 | 18.30 ± 0.35 |
MCHC [g/L] | 354.40 ± 6.45 | 350.56 ± 5.03 | 351.80 ± 5.73 | 345.00 ± 5.15* |
LEUKOCYTES [x 109/L] | 3.38 ± 0.42 | 3.74 ± 0.57 | 2.33 ± 0.43 | 2.96 ± 0.51* |
THROMBOCYTES [x 109/L] | 553.00 ± 38.24 | 515.56 ± 56.96 | 531.70 ± 33.13 | 507.33 ± 50.20 |
RETICULOCYTES [1/1] | 0.007 ± 0.001 | 0.007 ± 0.002 | 0.009 ± 0.002 | 0.009 ± 0.002 |
PT [s] | 16.13 ± 1.35 | 16.01 ± 0.91 | 15.05 ± 0.75 | 14.67 ± 0.88 |
APTT [s] | 30.43 ± 6.92 | 28.94 ± 6.15 | 26.82 ± 13.18 | 24.12 ± 7.12 |
* - statistically significant difference with p ≤ 0.05; Student's t-test
Table 13. Blood serum biochemical examinations - main groups
Parameter |
| GROUP / DOSE [mg/kg b.w.] |
| |||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |
| males | females |
| |||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | |
TOTAL PROTEIN [g/L] | 58.40 ±1.75 | 59.16 ±1.54 | 59.25 ±2.28 | 58.14 ±1.34 | 64. 59 ±2.70 | 61.98 ±3.05 | 63.84 ±2.53 | 60.57 ±3.91* |
ALBUMIN [g/L] | 31.90 ± 1.71 | 31.68 ±1.78 | 32.18 ± 1.94 | 30.93 ±1.46 | 36.02 ±1.69 | 34.83 ±1.57 | 35.49 ±1.81 | 33.58 ±3.36 |
GLOBULIN [g/L] | 26.50 ±2.18 | 27.48 ±2.20 | 27.07 ±2.07 | 27.21 ±2.43 | 28.57 ±1.35 | 27.15 ±1.70 | 28.35 ±2.05 | 26.99 ±1.25 |
A/G RATIO | 1.22 ±0.15 | 1.16 ±0.14 | 1.20 ±0.13 | 1.15 ±0.14 | 1.26 ±0.06 | 1.28 ±0.06 | 1.26 ±0.11 | 1.24 ±0.12 |
GLUCOSE [mmol/L] | 9.51 ±2.08 | 9.25 ±1.68 | 10.39 ± 1.75 | 9.93 ±2.40 | 8.56 ±2.36 | 8.10 ±1.38 | 7.48 ±0.91 | 7.41 ±0.74 |
CHOLESTEROL [mmol/L] | 1.76 ±0.24 | 1.70 ±0.14 | 1.84 ±0.30 | 1.86 ±0.31 | 1.53 ±0.36 | 1.32 ±0.28 | 1.36 ±0.39 | 1.27 ±0.28 |
HDL [mmol/L] | 1.15 ±0.15 | 1.14 ±0.11 | 1.24 ±0.21 | 1.22 ±0.19 | 1.08 ±0.25 | 0.93 ±0.18 | 0.94 ±0.29 | 0.85 ±0.21 |
LDL [mmol/L] | 0.38 ±0.14 | 0.56 ±0.30 | 0.39 ±0.13 | 0.36 ±0.09 | 0.42 ±0.27 | 0.45 ±0.27 | 0.42 ±0.19 | 0.46 ±0.21 |
UREA NITROGEN [mmol/L] | 4.95 ±0.75 | 4.91 ±0.52 | 4.75 ±0.58 | 5.66 ±0.33* | 5.25 ±1.19 | 5.97 ±0.86 | 5.52 ±0.60 | 6.03 ±1.17 |
CREATININE [μmol/L] | 28.20 ± 4.37 | 28.00 ± 2.11 | 28.90 ± 5.65 | 30.40 ±4.45 | 33.40 ± 3.31 | 31.20 ± 2.35 | 26.80 ±3.19* | 33.20 ±8.26 |
BILIRUBIN [μmol/L] | 3.78 ±0.46 | 3.78 ±0.64 | 3.82 ±0.63 | 3.98 ±0.48 | 4.77 ±2.03 | 4.64 ±1.58 | 4.71 ±1.27 | 5.82 ±3.08 |
SODIUM [mmol/L] | 144.30 ± 2.11 | 144.90 ± 1.60 | 146.40 ±1.78* | 146.00 ±1.63 | 145.10 ±1.52 | 144.30 ±1.34 | 145.70 ±1.16 | 144.70 ±1.25 |
POTASSIUM [mmol/L] | 4.21 ±0.33 | 4.16 ±0.21 | 4.21 ±0.30 | 4.38 ±0.32 | 3.91 ±0.48 | 4.06 ±0.42 | 4.03 ±0.36 | 4.26 ±0.36 |
CHLORIDES [mmol/L] | 104.30 ± 1.25 | 104.50 ±0.71 | 104.50 ±2.07 | 103.70 ±2.06 | 105.20 ±1.32 | 106.70 ±1.95 | 106.60 ±1.58 | 105.90 ±1.97 |
CALCIUM [mmol/L] | 2.39 ±0.08 | 2.43 ±0.07 | 2.48 ±0.08 | 2.42 ±0.07 | 2.55 ±0.07 | 2.48 ±0.08 | 2.55 ±0.06 | 2.44 ±0.08* |
INORGANIC PHOSPHOROUS [mmol/L] | 1.71 ±0.19 | 1.67 ±0.12 | 1.80 ±0.21 | 1.99 ±0.19* | 1.52 ±0.28 | 1.59 ±0.26 | 1.88 ±0.18* | 1.76 ±0.22 |
* - statistically significant difference with p ≤ 0.05; Dunnett's test
Table 14. Blood serum biochemical examinations - satellite groups
Parameter | GROUP / DOSE [mg/kg b.w.] | |||||
0SAT/0 | 3SAT/1000 | 0SAT/0 | 3SAT/1000 | |||
males | females | |||||
n=10 | n=10 | n=10 | n=9 | |||
TOTAL PROTEIN [g/L] | 63.18 ± 1.50 | 61.56 ± 2.72 | 63.59 ± 3.15 | 64.34 ± 2.08 | ||
ALBUMIN [g/L] | 33.67 ± 1.10 | 32.67 ± 1.50 | 35.78 ± 1.35 | 36.72 ± 1.23 | ||
GLOBULIN [g/L] | 29.51 ± 0.94 | 28.89 ± 1.48 | 27.81 ± 2.03 | 27.62 ± 1.30 | ||
A/G RATIO | 1.14 ± 0.05 | 1.13 ± 0.05 | 1.29 ± 0.07 | 1.33 ± 0.06 | ||
GLUCOSE [mmol/L] | 9.00 ± 1.00 | 8.84 ± 1.11 | 7.13 ± 0.93 | 7.30 ± 0.77 | ||
CHOLESTEROL [mmol/L] | 1.98 ± 0.24 | 1.99 ± 0.18 | 1.67 ± 0.36 | 1.90 ± 0.28 | ||
HDL [mmol/L] | 1.33 ± 0.15 | 1.38 ± 0.15 | 1.11 ± 0.20 | 1.22 ± 0.16 | ||
LDL [mmol/L] | 0.40 ± 0.06 | 0.36 ± 0.05 | 0.40 ± 0.15 | 0.41 ± 0.08 | ||
UREA NITROGEN [mmol/L] | 6.55 ± 0.93 | 6.31 ± 0.89 | 7.08 ± 1.24 | 7.33 ± 1.06 | ||
CREATININE [μmol/L] | 30.50 ± 6.04 | 29.00 ± 5.16 | 35.20 ± 5.49 | 38.11 ± 4.81 | ||
BILIRUBIN [μmol/L] | 4.18 ± 0.42 | 4.45 ± 0.53 | 4.46 ± 1.05 | 5.00 ± 0.92 | ||
SODIUM [mmol/L] | 136.80 ± 2.30 | 137.60 ± 0.97 | 143.90 ± 1.10 | 143.44 ± 0.88 | ||
POTASSIUM [mmol/L] | 3.82 ± 0.14 | 4.14 ± 0.24* | 3.83 ± 0.25 | 3.73 ± 0.39 | ||
CHLORIDES [mmol/L] | 97.40 ± 1.35 | 98.50 ± 1.27 | 104.80 ± 1.75 | 103.22 ± 0.97** | ||
CALCIUM [mmol/L] | 2.42 ± 0.05 | 2.40 ± 0.06 | 2.52 ± 0.06 | 2.47 ± 0.08 | ||
INORGANIC PHOSPHOROUS [mmol/L] | 1.87 ± 0.18 | 1.91 ± 0.20 | 1.68 ± 0.15 | 1.54 ± 0.27 | ||
x ± SD;
n - number of test animals
* - statistically significant difference with p ≤ 0.05; Student's t-test;
** - statistically significant difference with p ≤ 0.05; Mann-Whitney U test;
Table 15. Relative organ-to-body weight of internal organs [%] – main groups
Examined organ | GROUP / DOSE [mg/kg b.w./day] | |||||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |
males | females | |||||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | |
Brain with cerebellum | 0.493 ±0.029 | 0.491 ±0.038 | 0.471 ±0.038 | 0.515 ±0.045 | 0.795 ±0.044 | 0.757 ±0.061 | 0.782 ±0.068 | 0.774 ±0.049 |
Pituitary gland | 0.002 ±0.0004 | 0.002 ±0.001 | 0.002 ±0.001 | 0.002 ±0.0004 | 0.007 ±0.001 | 0.005* ±0.001 | 0.005* ±0.001 | 0.005* ±0.001 |
Thyroid | 0.006 ±0.001 | 0.006 ±0.001 | 0.006 ±0.001 | 0.006 ±0.001 | 0.008 ±0.002 | 0.007 ±0.002 | 0.008 ±0.001 | 0.008 ±0.001 |
Thymus | 0.084 ±0.022 | 0.097 ±0.018 | 0.082 ±0.015 | 0.073 ±0.015 | 0.124 ±0.020 | 0.130 ±0.015 | 0.126 ±0.022 | 0.117 ±0.025 |
Heart | 0.239 ±0.015 | 0.256 ±0.016 | 0.248 ±0.013 | 0.264* ±0.018 | 0.289 ±0.040 | 0.281 ±0.023 | 0.278 ±0.029 | 0.281 ±0.025 |
Liver | 2.700 ±0.279 | 2.978 ±0.291 | 2.922 ±0.212 | 2.977 ±0.223 | 2.980 ±0.227 | 2.863 ±0.295 | 2.709 ±0.345 | 2.862 ±0.178 |
Spleen | 0.131 ±0.015 | 0.151 ±0.011 | 0.143 ±0.019 | 0.156* ±0.025 | 0.206 ±0.014 | 0.202 ±0.022 | 0.206 ±0.038 | 0.208 ±0.026 |
Kidneys | 0.578 ±0.032 | 0.598 ±0.035 | 0.580 ±0.038 | 0.672* ±0.061 | 0.678 ±0.057 | 0.648 ±0.061 | 0.653 ±0.056 | 0.641 ±0.032 |
Adrenal glands | 0.017 ±0.002 | 0.017 ±0.003 | 0.016 ±0.001 | 0.016 ±0.002 | 0.038 ±0.004 | 0.035 ±0.004 | 0.035 ±0.005 | 0.035 ±0.005 |
Testicles | 0.865 ±0.054 | 0.873 ±0.064 | 0.869 ±0.053 | 0.936 ±0.106 | - | - | - | - |
Epididymides | 0.340 ±0.035 | 0.341 ±0.027 | 0.327 ±0.024 | 0.355 ±0.028 | - | - | - | - |
Prostate with seminal vesicles and coagulating glands | 0.461 ±0.083 | 0.487 ±0.056 | 0.443 ±0.083 | 0.464 ±0.047 | - | - | - | - |
Ovaries | - | - | - | - | 0.050 ±0.007 | 0.047 ±0.006 | 0.047 ±0.007 | 0.048 ±0.007 |
Uterus with cervix | - | - | - | - | 0.249 ±0.056 | 0.280 ±0.132 | 0.273 ±0.108 | 0.283 ±0.132 |
n- number of tested animals; x ± SD;
* - statistically significant difference with p ≤ 0.05, Dunnett's test
Table 16.Histopathological examination - main groups
Examined organ | Type of changes | Degree of severity (numerical score) | GROUP / DOSE [mg/kg b.w./day] / number of animals | |||||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |||
males | females | |||||||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | |||
Urinary bladder | hyperplasia urothelium, papillary, diffuse | minimal [1] | - | - | 2 | - | - | - | 4 | - |
slight [2] | - | - | - | - | - | - | 2 | - | ||
severe [4] | - | - | - | 10 | - | - | - | 10 | ||
proteinaceous plug | present [+] | 1
| - | - | - | - | - | - | - | |
infiltration/s, mononuclear cells | minimal [1] | - | - | 2
| 3
| - | - | - | 2 | |
slight [2] | - | - | - | - | - | - | - | 2 | ||
moderate [3] | - | - | - | - | - | - | - | 1 | ||
oedema | present [+] | - | - | - | - | 1 | - | - | 1 | |
ns – examination not scheduled, n – number of animals in group
Examined organ | Type of changes | Degree of severity (numerical score) | GROUP / DOSE [mg/kg b.w./day] / number of animals |
| ||||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |||
males | females |
| ||||||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | |||
Left kidney | hemorrhage/s | minimal [1] | 1 | 2 | 3 | 1 | 2 | 5 | 4 | 3 |
slight [2] | 3 | 7 | 6 | 7 | 4 | 4 | 2 | 3 | ||
moderate [3] | 6 | 1 | - | 2 | 4 | 1 | 2 | 1 | ||
renal tubules, basophilia | present [+] | 2 | 1 | 1 | 6 | 2 | - | - | 3 | |
renal tubules, degeneration, focal | present [+] | - | - | - | - | 1 | - | - | - | |
renal tubules, degeneration, diffuse | present [+] | 1 | 1 | 1 | - | - | - | 2 | - | |
infiltration/s, mononuclear cells, focal | minimal [1] | 2 | 2 | 4 | 2 | - | 1 | - | - | |
slight [2] | - | - | - | - | - | 1 | 1 | - | ||
ns – examination not scheduled, n – number of animals in group
Examined organ | Type of changes | Degree of severity (numerical score) | GROUP / DOSE [mg/kg b.w./day] / number of animals | |||||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |||
males | females | |||||||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | |||
Left kidney | cyst/s | present [+] | 4 | 2 | 1 | 1 | 1 | - | 2 | 2 |
mineralization, focal | present [+] | - | - | - | - | 7 | 5 | 7 | 5 | |
renal pelvis, urothelium, hyperplasia, papillary, diffuse | minimal [1] | - | - | - | 1 | - | - | - | 3 | |
slight [2] | - | - | - | 5 | - | - | - | 2 | ||
moderate [3] | - | - | - | 4 | - | - | - | - | ||
papilla, hyperplasia, epithelium, papillary diffuse | minimal [1] | - | - | - | 1 | - | - | - | 2 | |
slight [2] | - | - | - | 3 | - | - | - | 2 | ||
moderate [3] | - | - | - | 2 | - | - | - | 2 | ||
severe [4] | - | - | - | 4 | - | - | - | 1 | ||
fibrosis | present [+] | 1 | - | - | - | - | 1 | 1 | - | |
Right kidney | hemorrhage/s | minimal [1] | 1 | 1 | 1 | - | - | 1 | 1 | 3 |
slight [2] | 4 | 2 | 4 | 1 | 1 | 6 | 5 | 4 | ||
moderate [3] | 5 | 6 | 5 | 7 | 8 | 3 | 4 | 2 | ||
renal tubules, basophilia | present [+] | 3 | 2 | 2 | 8 | 2 | 1 | 1 | 1 | |
renal tubules, degeneration, diffuse | present [+] | - | 1 | - | 1 | - | - | 1 | - | |
infiltration/s, mononuclear cells, focal | minimal [1] | 4 | 3 | 2 | 2 | 2 | - | - | 3 | |
slight [2] | - | - | - | 2 | - | - | 1 | - | ||
n – number of animals in group
Examined organ | Type of changes | Degree of severity (numerical score) | GROUP / DOSE [mg/kg b.w./day] / number of animals | |||||||
0/0 | 1/62.5 | 2/250 | 3/1000 | 0/0 | 1/62.5 | 2/250 | 3/1000 | |||
males | females | |||||||||
n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | n=10 | |||
Right kidney
| cyst/s | present [+] | 3 | 7 | 3 | 3 | 2 | 5 | 4 | 3 |
papilla, atrophy | present [+] | - | - | 1 | - | - | - | - | - | |
fibrosis | present [+] | - | - | - | 2 | - | - | - | - | |
renal pelvis, urothelium, hyperplasia, papillary, diffuse | minimal [1] | - | - | - | 1 | - | - | - | 3 | |
slight [2] | - | - | - | 3 | - | - | - | 3 | ||
moderate [3] | - | - | - | 5 | - | - | - | 1 | ||
papilla, hyperplasia, epithelium, papillary diffuse | minimal [1] | - | - | - | 5 | - | - | - | 3 | |
slight [2] | - | - | - | 1 | - | - | - | 1 | ||
moderate [3] | - | - | - | 5 | - | - | - | 1 | ||
severe [4] | - | - | - | 2 | - | - | - | - | ||
mineralization, focal | present [+] | - | 1 | - | - | - | 5 | 6 | 8 | |
subscapular, necrosis, focal | present [+] | - | - | 1 | - | - | - | - | - | |
fibrosis | present [+] | - | - | - | - | - | - | - | 1 | |
n – number of animals in group
Table 17. Histopathological examination - satellite groups
Examined organ | Type of changes | Degree of severity (numerical score) | GROUP / DOSE [mg/kg b.w./day] / number of animals | |||
0SAT/0 | 3SAT/1000 | 0SAT/0 | 3SAT/1000 | |||
males | females | |||||
n=10 | n=10 | n=10 | n=10 | |||
Urinary bladder | hyperplasia urothelium, papillary, diffuse | minimal [1] | - | - | - | 3 |
slight [2] | - | 7 | - | 5 | ||
moderate [3] | - | 3 | - | - | ||
infiltration/s, mononuclear cells | minimal [1] | - | 1 | - | - | |
oedema | present [+] | - | - | - | 1 | |
Left kidney | hemorrhage/s | minimal [1] | 1 | 3 | 6 | 2 |
slight [2] | 4 | 4 | 1 | 5 | ||
moderate [3] | 3 | 1 | 1 | 2 | ||
renal tubules, basophilia | present [+] | - | 3 | - | 1 | |
renal tubules, degeneration, focal | present [+] | - | 1 | 3 | - | |
renal tubules, degeneration, diffuse | minimal [1] | 3 | - | - | - | |
infiltration/s, mononuclear cells, focal | minimal [1] | 5 | 2 | 1 | - | |
cyst/s | present [+] | 1 | 3 | - | 3 | |
papilla, atrophy | present [+] | - | - | - | 1 | |
n – number of animals in group
Examined organ | Type of changes | Degree of severity (numerical score) | GROUP / DOSE [mg/kg b.w./day] / number of animals | |||
0SAT/0 | 3SAT/1000 | 0SAT/0 | 3SAT/1000 | |||
males | females | |||||
n=10 | n=10 | n=10 | n=10 | |||
Left kidney | renal tubule, dilation | present [+] | 1 | - | 2 | 1 |
mineralization, focal | present [+] | - | - | 7 | 6 | |
hyperplasia, papillary, diffuse, epithelium papilla | minimal [1] | - | 6 | - | 3 | |
slight [2] | - | 1 | - | 4 | ||
moderate [3] | - | - | - | 1 | ||
renal pelvis, urothelium, hyperplasia, papillary, diffuse | minimal [1] | - | 3 | - | 4 | |
slight [2] | - | 1 | - | 2 | ||
glomerulus, epithelium hyperplasia | present [+] | 1 | - | - | - | |
Right kidney
| hemorrhage/s | minimal [1] | 2 | 2 | 1 | 1 |
slight [2] | 3 | 4 | 8 | 4 | ||
moderate [3] | 5 | 3 | 1 | 4 | ||
renal tubules, basophilia | present [+] | 1 | 4 | 1 | 1 | |
renal tubules, degeneration, focal | present [+] | - | - | 1 | - | |
renal tubules, degeneration, diffuse | present [+] | 2 | - | - | - | |
infiltration/s, mononuclear cells, focal | minimal [1] | 3 | 5 | 2 | - | |
slight [2] | 1 | - | - | - | ||
moderate [3] | 1 | - | - | - | ||
cyst/s | present [+] | 5 | 6 | 2 | 2 | |
pelvis dilation | present [+] | - | - | - | - | |
renal tubule, dilation | present [+] | 1 | - | 1 | 1 | |
mineralization, focal | present [+] | - | - | 7 | 9 | |
hyperplasia, papillary, diffuse, epithelium papilla | minimal [1] | - | 4 | - | 1 | |
slight [2] | - | 3 | - | 2 | ||
moderate [3] | - | - | - | 2 | ||
renal pelvis, urothelium, hyperplasia, papillary, diffuse | minimal [1] | - | 3 | - | 6 | |
slight [2] | - | 4 | - | 1 | ||
glomerulus, epithelium hyperplasia | present [+] | 1 | - | - | - | |
n – number of animals in group
Table 5: Main study. Clinical signs - adult animals
Clinical signs |
Group / sex / number of animals / computer numbers of animals |
|||||||
0 |
1 |
2 |
3 |
|||||
males |
females |
males |
females |
males |
females |
males |
females |
|
Test animals |
10 |
12 |
10 |
12 |
10 |
12 |
10 |
12 |
Animals with clinical signs |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
Animals with ophthalmic changes |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Death of animals |
- |
- |
- |
- |
- |
- |
- |
- |
SKIN AND COAT |
||||||||
Thinning of hair all over the body |
- |
- |
- |
1 [16] |
- |
- |
- |
- |
[ ] computer numbers of animals
Table 6. Main study. Clinical signs - satellite groups
Clinical signs |
Group / sex / number of animals / computer numbers of animals |
|||
0 SAT |
3 SAT |
|||
males |
females |
males |
females |
|
Test animals |
10 |
10 |
10 |
10 |
Animals with clinical signs |
0 |
0 |
0 |
0 |
Animals with ophthalmic changes |
0 |
0 |
0 |
0 |
Death of animals |
- |
- |
- |
- |
Table 7. VL3. Main study. Open field observations – males
Parameter |
Group |
|||
0 n = 5 |
1 n = 5 |
2 n = 5 |
3 n = 5 |
|
involuntary clonic movement |
no changes |
no changes |
no changes |
no changes |
nvoluntary tonic movement |
no changes |
no changes |
no changes |
no changes |
gait |
no changes |
no changes |
no changes |
no changes |
arousal |
[4]1, [5]1 |
[3]4 |
[5]4 |
no changes |
stereotypies |
no changes |
no changes |
no changes |
no changes |
number of defecations |
2.20±1.30 |
2.40±2.30 |
1.00±1.73 |
2.80±1.92 |
number of urinations |
2.00±0.71 |
1.40±0.89 |
2.40±0.55 |
1.80±1.10 |
distance 0-3 min [m] |
69.26±33.14 |
100.28±38.03 |
157.78±52.14* |
119.38±43.20 |
number of rearings 0-3 min |
5.60±5.13 |
10.00±11.40 |
13.60±8.20 |
7.40±3.51 |
x± SD
n–number of test animals
*-statistically significant difference at p ≤ 0.05-Dunnett’s test
[ ] - computer numbers of animals with changes
Classification of changes:
1 - slight decrease (a rat sometimes does not move, but exploratory activity is observed)
2 - moderate decrease (a rat is slightly numb; only the head moves)
3 - strong decrease (paralysis, coma)
4 - slight increase (a rat is slightly excited or tense; it suddenly moves forward and stays without motion)
5 - strong increase (a rat is very vigilant; it runs and suddenly moves from its place
Table 8. VL3. Main study. Open field observations – females
Parameter |
Group |
|||
0 n = 5 |
1 n = 5 |
2 n = 5 |
3 n = 5 |
|
involuntary clonic movement |
no changes |
no changes |
no changes |
no changes |
nvoluntary tonic movement |
no changes |
no changes |
no changes |
no changes |
gait |
no changes |
no changes |
no changes |
no changes |
arousal |
no changes |
no changes |
[12]4, [15]4, [16]4 |
[11]1, [12]4, [14]4 |
stereotypies |
no changes |
no changes |
no changes |
no changes |
number of defecations |
2.80±2.17 |
1.80±2.49 |
2.20±2.05 |
1.00±1.73 |
number of urinations |
2.60±1.52 |
4.20±0.84 |
4.00±1.73 |
2.20±1.64 |
distance 0-3 min [m] |
109.44±18.48 |
106.48±12.37 |
162.06±40.87* |
125.40±39.00 |
number of rearings 0-3 min |
13.60±4.22 |
12.80±2.77 |
18.60±8.20 |
19.20±7.09 |
Table 9. VL3. Main study. Open field observations–males (satellite groups)–measurement 1
Parameter |
Group |
|
0 SAT n = 5 |
3 SAT n = 5 |
|
involuntary clonic movement |
no changes |
no changes |
nvoluntary tonic movement |
no changes |
no changes |
gait |
no changes |
no changes |
arousal |
[34]4 |
no changes |
stereotypies |
no changes |
no changes |
number of defecations |
2.20±1.79 |
2.20±1.92 |
number of urinations |
1.60±1.52 |
1.60±0.55 |
distance 0-3 min [m] |
96.42±28.44 |
96.06±17.15 |
number of rearings 0-3 min |
8.20±1.92 |
11.60±3.91 |
Table 10. VL3. Main study. Open field observations - females (satellite groups)–measurement 1
Parameter |
Group |
|
0 SAT n = 5 |
3 SAT n = 5 |
|
involuntary clonic movement |
no changes |
no changes |
nvoluntary tonic movement |
no changes |
no changes |
gait |
no changes |
no changes |
arousal |
[44]4 |
no changes |
stereotypies |
no changes |
no changes |
number of defecations |
0.00±0.00 |
1.00±1.41 |
number of urinations |
1.60±0.55 |
1.20±0.84 |
distance 0-3 min [m] |
106.62±17.52 |
115.86±4.27 |
number of rearings 0-3 min |
15.00±8.34 |
19.80±6.69 |
Table 11. VL3. Main study. Open field observations – males (satellite groups) – measurement 2
Parameter |
Group |
|
0 SAT n = 5 |
3 SAT n = 5 |
|
involuntary clonic movement |
no changes |
no changes |
nvoluntary tonic movement |
no changes |
no changes |
gait |
no changes |
no changes |
arousal |
[31]2, [32]2, [34]1 |
[31]2, [32]1, [33]1 [34]2, [35]2 |
stereotypies |
no changes |
no changes |
number of defecations |
0.80±1.10 |
2.20±2.28 |
number of urinations |
1.60±0.55 |
2.80±0.84* |
distance 0-3 min [m] |
64.82±24.68 |
44.40±19.49 |
number of rearings 0-3 min |
2.80±3.11 |
0.60±1.34 |
Table 12. VL3. Main study. Open field observations – females (satellite groups) – measurement 2
Parameter |
Group |
|
0 SAT n = 5 |
3 SAT n = 5 |
|
involuntary clonic movement |
no changes |
no changes |
nvoluntary tonic movement |
no changes |
no changes |
gait |
no changes |
no changes |
arousal |
[42]1, [44]2 |
[42]4, [43]1, [45]1
|
stereotypies |
no changes |
no changes |
number of defecations |
1.40±1.95 |
0.80±1.79 |
number of urinations |
2.80±2.95 |
2.60±1.52 |
distance 0-3 min [m] |
96.78±55.48 |
111.34±68.45 |
number of rearings 0-3 min |
3.80±2.68 |
9.60±5.73 |
Table 13. VL3. Main study. Reactions to stimuli
PAIN RESPONSE LATENCY [s] |
Group |
|||
0 n = 5 |
1 n = 5 |
2 n = 5 |
3 n = 5 |
|
Males |
3.44±0.27 |
3.62 ±0.19 |
3.58±0.45 |
3.40±0.32 |
Females |
2.94±0.40 |
3.16±0.30 |
3.06±0.36 |
3.24±0.09 |
No changes on approach response, touch response, sound response, pinna reflex.
Table 14. VL3. Main study. Reactions to stimuli (satellite groups)
PAIN RESPONSE LATENCY [s] |
Group |
|
0 SAT n = 5 |
3 SAT n = 5 |
|
Males – measurement 1 |
3.26±0.11 |
3.42±0.38 |
Females - measurement 1 |
3.24±0.29 |
2.42±0.23* |
Males – measurement 2 |
2.92±0.39 |
2.98±0.16 |
Females – measurement 2 |
2.54±0.68 |
2.40±0.72 |
No changes on approach response, touch response, sound response, pinna reflex.
x± SD
n–number of test animals
*– statistically significant difference at p ≤ 0.05- Student's t-test
Table 15. VL3. Main study. Limb grip strength
|
Group |
||||
Parameter |
0 n = 5 |
1 n = 5 |
2 n = 5 |
3 n = 5 |
|
Males |
forelimb grip strength [p] |
1208.20± 114.02 |
1234.50± 241.70 |
1030.50± 46.81 |
1221.60± 106.38 |
hindlimb grip strength [p] |
176.60± 33.96 |
167.60± 42.67 |
134.70± 45.15 |
140.30± 28.20 |
|
Females |
forelimb grip strength [p] |
1194.50± 241.89 |
1159.20± 138.32 |
1134.20± 61.33 |
1263.10± 79.53 |
hindlimb grip strength [p] |
187.80± 47.64 |
182.60± 54.94 |
193.10± 47.25 |
180.60± 7.67 |
|
Table 16. VL3. Main study. Limb grip strength – satellite groups
|
Group |
||
Parameter |
0 SAT n = 5 |
3 SAT n = 5 |
|
Males – measurement 1 |
forelimb grip strength [p] |
1447.00±143.15 |
1422.30±116.27 |
hindlimb grip strength [p] |
174.90±54.84 |
186.00±46.09 |
|
Females - measurement 1 |
forelimb grip strength [p] |
1176.70±169.79 |
1206.30±110.01 |
hindlimb grip strength [p] |
187.80±47.64 |
160.60±39.50 |
|
Males – measurement 2 |
forelimb grip strength [p] |
1322.40±368.82 |
1423.30±185.26 |
hindlimb grip strength [p] |
175.60±25.91 |
182.00±11.09 |
|
Females – measurement 2 |
forelimb grip strength [p] |
1072.60±136.42 |
1357.40±195.27* |
hindlimb grip strength [p] |
133.20±29.48 |
142.20±29.91 |
|
x± SD
n–number of test animals
*- statistically significant difference at p ≤ 0.05- Student's t-test
Table 17. VL3. Main study. Results of haematological and coagulological examination – males
Parameter |
Group |
|||
0 n= 10 |
1 n = 10 |
2 n = 10 |
3 n = 10 |
|
HEMOGLOBIN (g/L) |
154.30 ± 5.46 |
153.40 ± 2.72 |
153.70 ± 4.72 |
153.30 ± 5.70 |
HEMATOCRIT (1/1) |
0.43 ± 0.02 |
0.42 ± 0.01 |
0.42 ± 0.02 |
0.42 ± 0.02 |
ERYTHROCYTES (x 1012/L) |
8.98 ± 0.26 |
8.98 ± 0.44 |
8.96 ± 0.28 |
8.92 ± 0.31 |
MCV (fL) |
47.70 ± 1.38 |
47.17 ± 1.46 |
47.26 ± 1.43 |
46.99 ± 1.72 |
MCH (pg) |
17.17 ± 0.42 |
17.12 ± 0.65 |
17.17 ± 0.40 |
17.17 ± 0.40 |
MCHC (g/L) |
360.10 ± 8.86 |
362.20 ± 6.39 |
363.50 ± 6.92 |
366.20 ± 5.92 |
THROMBOCYTES (x 109/L) |
667.90 ± 68.70 |
644.70 ± 55.69 |
648.50 ± 54.15 |
622.60 ± 55.46 |
LEUKOCYTES (x 109/L) |
4.32 ± 1.24 |
4.07 ± 0.91 |
5.12 ± 0.94 |
3.77 ± 0.84 |
RETICULOCYTES (1/1) |
0.019 ± 0.006 |
0.017 ± 0.004 |
0.015 ± 0.005 |
0.020 ± 0.005 |
HEINZ BODIES (‰) |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
PT (s) |
12.78 ± 0.69 |
12.90 ± 0.87 |
12.36 ± 0.98 |
11.82 ± 1.01 |
APTT (s) |
52.40 ± 2.41 |
51.40 ± 4.04 |
50.20 ± 7.85 |
47.40 ± 7.16 |
x± SD;
n - number of test animals
Table 18. VL3. Main study. Results of hematological and coagulological examination – females
Parameter |
Group |
|||
0 n= 10 |
1 n = 10 |
2 n = 10 |
3 n = 10 |
|
HEMOGLOBIN (g/L) |
147.40 ± 4.65 |
141.08 ± 5.79* |
147.08 ± 4.32 |
150.09 ± 6.67 |
HEMATOCRIT (1/1) |
0.41 ± 0.01 |
0.39 ± 0.02* |
0.40 ± 0.02 |
0.41 ± 0.02 |
ERYTHROCYTES (x 1012/L) |
7.85 ± 0.33 |
7.37 ± 0.34* |
7.67 ± 0.28 |
8.06 ± 0.55 |
MCV (fL) |
51.76 ± 1.30 |
52.24 ± 1.64 |
52.38 ± 2.24 |
50.82 ± 1.08 |
MCH (pg) |
18.81 ± 0.52 |
19.17 ± 0.45 |
19.20 ± 0.71 |
18.65 ± 0.59 |
MCHC (g/L) |
363.20 ± 5.43 |
366.83 ± 4.73 |
366.50 ± 6.36 |
367.45 ± 5.37 |
THROMBOCYTES (x 109/L) |
724.40 ± 53.19 |
664.42 ± 87.80 |
693.83 ± 43.16 |
756.36 ± 37.99 |
LEUKOCYTES (x 109/L) |
4.75 ± 1.23 |
3.94 ± 1.16 |
4.03 ± 1.53 |
4.82 ± 0.97 |
RETICULOCYTES (1/1) |
0.018 ± 0.004 |
0.017 ± 0.004 |
0.015 ± 0.004 |
0.017 ± 0.005 |
HEINZ BODIES (‰) |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
PT (s) |
9.66 ± 0.13 |
9.64 ± 0.05 |
9.94 ± 0.45 |
9.64 ± 0.09 |
APTT (s) |
47.80 ± 8.38 |
53.60 ± 1.52 |
52.80 ± 1.10 |
53.80 ± 2.59 |
x± SD;
n - number of test animals
* -statistically significant difference with p ≤ 0.05; Dunnett's test;
Table 19. VL3. Main study. Results of hematological and coagulological examination – males (satellite groups)
Parameter |
GROUP |
||
0 SAT n=10 |
3 SAT n=10 |
||
HEMOGLOBIN |
g/L |
149.00±4.92 |
152.30±4.76 |
HEMATOCRIT |
1/1 |
0.42 ± 0.02 |
0.42 ±0.02 |
ERYTHROCYTES |
x 1012/L |
8.80土0.41 |
9.02 ± 0.27 |
MCV |
fL |
47.23± 1.67 |
46.62± 1.33 |
MCH |
pg |
16.94± 0.53 |
16.90± 0.33 |
MCHC |
g/L |
358.70± 7.07 |
362.50±6.57 |
THROMBOCYTES |
x 109/L |
609.40± 70.84 |
662.50± 22.11* |
LEUKOCYTES |
x 109/L |
3.45土0.66 |
3.47 ±0.96 |
RETICULOCYTES |
1/1 |
0.020± 0.003 |
0.020± 0.003 |
HEINZ BODIES |
%〇 |
0.00土0.00 |
0.00 ±0.00 |
|
|
0 SAT |
3 SAT |
|
|
n=5 |
n=5 |
PT |
s |
12.12± 0.97 |
12.24± 0.65 |
APTT |
s |
50.80± 5.12 |
54.00± 3.08 |
Table 20. VL3. Main study. Results of hematological and coagulological examination – females (satellite groups)
Parameter |
GROUP |
||
0 SAT n=10 |
3 SAT n=10 |
||
HEMOGLOBIN |
g/L |
145.40± 5.48 |
149.00±6.13 |
HEMATOCRIT |
1/1 |
0.41 ± 0.02 |
0.41 ± 0.02 |
ERYTHROCYTES |
x 1012/L |
8.01 ± 0.37 |
8.12 ±0.35 |
MCV |
fL |
51.28± 1.95 |
50.53± 1.88 |
MCH |
pg |
18.17± 0.47 |
18.36± 0.50 |
MCHC |
g/L |
354.40± 7.85 |
363.70±6.27* |
THROMBOCYTES |
x 109/L |
614.80± 46.17 |
672.60± 64.14* |
LEUKOCYTES |
x 109/L |
2.63 ± 0.83 |
2.31 ± 0.56 |
RETICULOCYTES |
1/1 |
0.018±0.003 |
0.019± 0.002 |
HEINZ BODIES |
%〇 |
0.00 ± 0.00 |
0.00 ±0.00 |
|
|
0 SAT |
3 SAT |
|
|
n=5 |
n=5 |
PT |
s |
9.96 ± 0.21 |
10.04± 0.48 |
APTT |
s |
51.80± 3.90 |
48.60±2.51 |
Table 21. VL3. Main study. Results of hematological examination - males Percentage content of leukocytes (leukocytogram)
Parameter |
GROUP |
||||
0 n=10 |
1 n=10 |
2 n=10 |
3 n=10 |
||
NEUTROCYTES |
1/1 |
0.23土0.06 |
0.22土0.09 |
0.23土0.06 |
0.22土0.05 |
EOSINOCYTES |
1/1 |
0.01 ± 0.01 |
0.00土0.01 |
0.00土0.00 |
0.00土0.00 |
LYMPHOCYTES |
1/1 |
0.76土0.06 |
0.77土0.09 |
0.77土0.06 |
0.78土0.05 |
MONOCYTES |
1/1 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
OTHER CELLS |
1/1 |
0.00土0.01 |
0.00土0.00 |
0.00土0.01 |
0.00土0.00 |
Table 22. VL3. Main study. Results of hematological examination - females Percentage content of leukocytes (leukocytogram)
Parameter |
GROUP |
||||
0 n=10 |
1 n=12 |
2 n=12 |
3 n=11 |
||
NEUTROCYTES |
1/1 |
0.46土0.13 |
0.48土0.06 |
0.38土0.17 |
0.48土0.11 |
EOSINOCYTES |
1/1 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
LYMPHOCYTES |
1/1 |
0.54土0.13 |
0.51土0.06 |
0.61土0.17 |
0.52土0.11 |
MONOCYTES |
1/1 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
OTHER CELLS |
1/1 |
0.00土0.01 |
0.00土0.00 |
0.01土0.01 |
0.00土0.00 |
Table 23. VL3. Main study. Results of hematological examination - (satellite groups) Percentage content of leukocytes (leukocytogram)
Parameter |
GROUP (males) |
GROUP (females) |
|||
0 SAT n=10 |
3 SAT n=10 |
0 SAT n=10 |
3 SAT n=10 |
||
NEUTROCYTES |
1/1 |
0.19土0.05 |
0.21土0.04 |
0.14土0.06 |
0.16土0.05 |
EOSINOCYTES |
1/1 |
0.01 ± 0.01 |
0.01土0.01 |
0.01土0.01 |
0.00土0.00 |
LYMPHOCYTES |
1/1 |
0.81 ± 0.05 |
0.79土0.04 |
0.85土0.06 |
0.83土0.05 |
MONOCYTES |
1/1 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
OTHER CELLS |
1/1 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.01土0.01 |
Table 24. VL3. Main study. Results of hematological studies - bone marrow examination Erythrocyte system - males
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
PROERYTHROBLASTS 1/1 |
0.013± 0.007 |
0.011 ±0.002 |
0.013±0.004 |
0.013±0.004 |
BASOPHILIC ERYTHROBLASTS 1/1 |
0.015± 0.003 |
0.016± 0.010 |
0.018±0.006 |
0.014±0.004 |
POLYCHROMATOPHILIC ERYTHROBLASTS 1/1 |
0.170± 0.022 |
0.156± 0.006 |
0.150±0.020 |
0.154±0.011 |
ORTHOCHROMATIC ERYTHROBLASTS 1/1 |
0.206± 0.018 |
0.223± 0.035 |
0.219±0.019 |
0.208 ± 0.021 |
TOTAL 1/1 |
0.405± 0.037 |
0.406± 0.034 |
0.401 ±0.030 |
0.390±0.033 |
Table 25. VL3. Main study. Results of hematological studies - bone marrow examination Erythrocyte system - females
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
PROERYTHROBLASTS 1/1 |
0.012± 0.003 |
0.010± 0.002 |
0.014±0.007 |
0.011 ±0.003 |
BASOPHILIC ERYTHROBLASTS 1/1 |
0.014± 0.004 |
0.014± 0.005 |
0.014±0.009 |
0.014±0.004 |
POLYCHROMATOPHILIC ERYTHROBLASTS 1/1 |
0.165± 0.019 |
0.170± 0.016 |
0.156±0.029 |
0.166±0.011 |
ORTHOCHROMATIC ERYTHROBLASTS 1/1 |
0.218± 0.028 |
0.239± 0.013 |
0.230±0.033 |
0.240±0.010 |
TOTAL 1/1 |
0.410± 0.043 |
0.432± 0.018 |
0.414±0.063 |
0.430 ± 0.020 |
Table 26. VL3. Main study. Results of hematological studies - bone marrow examination Erythrocyte system - (satellite groups)
Parameter |
|
GROUP (males) |
GROUP (females) |
||
|
0 SAT n=5 |
3 SAT n=5 |
0 SAT n=5 |
3 SAT n=5 |
|
PROERYTHROBLASTS |
1/1 |
0.011 ± 0.003 |
0.006 ± 0.002* |
0.010± 0.002 |
0.010± 0.004 |
BASOPHILIC ERYTHROBLASTS |
1/1 |
0.016± 0.005 |
0.015± 0.004 |
0.015± 0.005 |
0.013± 0.005 |
POLYCHROMATOPHILIC ERYTHROBLASTS |
1/1 |
0.159± 0.024 |
0.158± 0.014 |
0.152± 0.019 |
0.175± 0.015 |
ORTHOCHROMATIC ERYTHROBLASTS |
1/1 |
0.210± 0.024 |
0.213± 0.031 |
0.211 ± 0.027 |
0.206± 0.015 |
TOTAL |
1/1 |
0.397± 0.038 |
0.392± 0.038 |
0.388± 0.039 |
0.404± 0.026 |
Table 27. VL3. Main study. Results of hematological studies - bone marrow examination Leukocyte system – males
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
MYELOBLASTS 1/1 |
0.011 ± 0.007 |
0.016± 0.008 |
0.012±0.005 |
0.014±0.004 |
PROMYELOCYTES 1/1 |
0.022± 0.006 |
0.030± 0.008 |
0.025±0.003 |
0.022 ± 0.002 |
ORTHOCHROMATOPHILIC MYELOCYTES 1/1 |
0.035± 0.005 |
0.033± 0.008 |
0.030±0.005 |
0.031 ±0.003 |
ACIDOPHILIC MYELOCYTES 1/1 |
0.003± 0.001 |
0.004± 0.001 |
0.003±0.001 |
0.003±0.001 |
ORTHOCHROMATOPHILIC METAMYELOCYTES 1/1 |
0.044± 0.005 |
0.042± 0.002 |
0.046 ± 0.005 |
0.045±0.003 |
ACIDOPHILIC METAMYELOCYTES 1/1 |
0.010± 0.004 |
0.008± 0.005 |
0.014±0.005 |
0.010±0.005 |
ROD NEUTROPHILS 1/1 |
0.042± 0.008 |
0.043±0.012 |
0.038±0.012 |
0.042±0.012 |
ROD EOSINOPHILS 1/1 |
0.006± 0.002 |
0.006± 0.002 |
0.006 ± 0.004 |
0.003±0.001 |
FILAMENTED NEUTROPHILS 1/1 |
0.169± 0.027 |
0.147± 0.019 |
0.169±0.035 |
0.170±0.027 |
FILAMENTED EOSINOPHILS 1/1 |
0.018± 0.009 |
0.018± 0.009 |
0.025±0.010 |
0.011 ±0.003 |
BASOPHILS 1/1 |
0.002± 0.001 |
0.003± 0.001 |
0.002±0.001 |
0.003±0.001 |
TOTAL 1/1 |
0.362± 0.027 |
0.349± 0.019 |
0.369 ± 0.040 |
0.353±0.029 |
Leukocyte system/erythrocyte system quantitative ratio |
0.903± 0.130 |
0.866± 0.092 |
0.929±0.160 |
0.916±0.158 |
Table 28. VL3. Main study. Results of hematological studies - bone marrow examination Leukocyte system - females
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
MYELOBLASTS 1/1 |
0.011 ± 0.003 |
0.014± 0.002 |
0.013±0.003 |
0.014±0.004 |
PROMYELOCYTES 1/1 |
0.023± 0.007 |
0.022± 0.004 |
0.022 ± 0.005 |
0.026 ± 0.007 |
ORTHOCHROMATOPHILIC MYELOCYTES 1/1 |
0.036± 0.006 |
0.034± 0.006 |
0.030±0.005 |
0.036±0.007 |
ACIDOPHILIC MYELOCYTES 1/1 |
0.004± 0.002 |
0.004± 0.003 |
0.003±0.001 |
0.004 ± 0.002 |
ORTHOCHROMATOPHILIC METAMYELOCYTES 1/1 |
0.046± 0.006 |
0.048± 0.014 |
0.042 ± 0.008 |
0.051 ±0.014 |
ACIDOPHILIC METAMYELOCYTES 1/1 |
0.014± 0.004 |
0.013± 0.004 |
0.014±0.010 |
0.009±0.003 |
ROD NEUTROPHILS 1/1 |
0.058± 0.008 |
0.052± 0.020 |
0.063±0.013 |
0.050±0.019 |
ROD EOSINOPHILS 1/1 |
0.004± 0.003 |
0.005± 0.003 |
0.009±0.009 |
0.003±0.001 |
FILAMENTED NEUTROPHILS 1/1 |
0.171 ± 0.011 |
0.170± 0.021 |
0.173±0.037 |
0.180±0.019 |
FILAMENTED EOSINOPHILS 1/1 |
0.026± 0.008 |
0.019± 0.010 |
0.018±0.011 |
0.022±0.010 |
BASOPHILS 1/1 |
0.002± 0.000 |
0.002± 0.001 |
0.002±0.001 |
0.002±0.001 |
TOTAL 1/1 |
0.394± 0.018 |
0.384± 0.019 |
0.388±0.043 |
0.398±0.017 |
Leukocyte system/erythrocyte system quantitative ratio |
0.973± 0.140 |
0.891 ± 0.078 |
0.946 ± 0.086 |
0.928±0.083 |
Table 29. VL3. Main study. Results of hematological studies - bone marrow examination Leukocyte system - (satellite groups)
Parameter |
GROUP (males) |
GROUP (females) |
||
0 SAT n=5 |
3 SAT n=5 |
0 SAT n=5 |
3 SAT n=5 |
|
MYELOBLASTS 1/1 |
0.015± 0.007 |
0.012± 0.004 |
0.012± 0.003 |
0.013± 0.003 |
PROMYELOCYTES 1/1 |
0.024± 0.006 |
0.023± 0.008 |
0.024± 0.005 |
0.023± 0.005 |
ORTHOCHROMATOPHILIC MYELOCYTES 1/1 |
0.027± 0.007 |
0.028± 0.006 |
0.033± 0.005 |
0.032± 0.005 |
ACIDOPHILIC MYELOCYTES 1/1 |
0.004± 0.002 |
0.003± 0.001 |
0.003± 0.001 |
0.004± 0.002 |
ORTHOCHROMATOPHILIC METAMYELOCYTES 1/1 |
0.046± 0.018 |
0.050± 0.012 |
0.046± 0.010 |
0.046± 0.007 |
ACIDOPHILIC METAMYELOCYTES 1/1 |
0.013± 0.008 |
0.007± 0.003 |
0.018± 0.013 |
0.020± 0.010 |
ROD NEUTROPHILS 1/1 |
0.043± 0.009 |
0.040± 0.013 |
0.049± 0.012 |
0.048± 0.008 |
ROD EOSINOPHILS 1/1 |
0.004± 0.002 |
0.002± 0.001 |
0.005± 0.004 |
0.006± 0.004 |
FILAMENTED NEUTROPHILS 1/1 |
0.163± 0.037 |
0.165± 0.019 |
0.150± 0.017 |
0.146± 0.012 |
FILAMENTED EOSINOPHILS 1/1 |
0.029± 0.005 |
0.018± 0.008* |
0.022± 0.010 |
0.026± 0.005 |
BASOPHILS 1/1 |
0.002± 0.001 |
0.002± 0.001 |
0.003± 0.001 |
0.002± 0.001 |
TOTAL 1/1 |
0.370± 0.023 |
0.351 ± 0.018 |
0.364± 0.029 |
0.366± 0.026 |
Leukocyte system/erythrocyte system quantitative ratio |
0.944± 0.143 |
0.905± 0.124 |
0.948± 0.131 |
0.911 ± 0.102 |
Table 30. VL3. Main study. Results of hematological studies - bone marrow examination Different cells - males
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
LYMPHOCYTES 1/1 |
0.187± 0.031 |
0.204± 0.031 |
0.180±0.019 |
0.217±0.023 |
MONOCYTES 1/1 |
0.011 ± 0.003 |
0.009± 0.007 |
0.012±0.003 |
0.009±0.005 |
PLASMOCYTES 1/1 |
0.005± 0.003 |
0.005± 0.003 |
0.008±0.005 |
0.003±0.001 |
MEGAKARYOCYTES 1/1 |
0.003± 0.001 |
0.004± 0.003 |
0.003±0.001 |
0.002±0.001 |
OTHER CELLS 1/1 |
0.026± 0.005 |
0.023± 0.008 |
0.027 ± 0.006 |
0.026 ± 0.006 |
TOTAL 1/1 |
0.233± 0.031 |
0.245± 0.034 |
0.230 ± 0.021 |
0.257 ± 0.023 |
Table 31. VL3. Main study. Results of hematological studies - bone marrow examination Different cells - females
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
LYMPHOCYTES 1/1 |
0.148± 0.021 |
0.141 ± 0.008 |
0.158±0.105 |
0.130±0.019 |
MONOCYTES 1/1 |
0.013± 0.005 |
0.012± 0.003 |
0.013±0.007 |
0.012±0.005 |
PLASMOCYTES 1/1 |
0.005± 0.004 |
0.004± 0.002 |
0.003±0.001 |
0.003±0.001 |
MEGAKARYOCYTES 1/1 |
0.002± 0.001 |
0.003± 0.001 |
0.003±0.001 |
0.002±0.001 |
OTHER CELLS 1/1 |
0.028± 0.006 |
0.024± 0.004 |
0.022 ± 0.005 |
0.025±0.008 |
TOTAL 1/1 |
0.196± 0.026 |
0.184± 0.010 |
0.199±0.101 |
0.172±0.009 |
Table 32. VL3. Main study. Results of hematological studies - bone marrow examination Different cells - (satellite groups)
Parameter |
GROUP (males) |
GROUP (females) |
|||
0 SAT n=5 |
3 SAT n=5 |
0 SAT n=5 |
3 SAT n=5 |
||
LYMPHOCYTES |
1/1 |
0.187± 0.016 |
0.211 ± 0.022 |
0.210± 0.036 |
0.185± 0.024 |
MONOCYTES |
1/1 |
0.009± 0.004 |
0.010± 0.004 |
0.009± 0.002 |
0.012± 0.009 |
PLASMOCYTES |
1/1 |
0.004± 0.002 |
0.004± 0.002 |
0.003± 0.001 |
0.005± 0.004 |
MEGAKARYOCYTES |
1/1 |
0.003± 0.001 |
0.004± 0.002 |
0.003± 0.001 |
0.004± 0.002 |
OTHER CELLS |
1/1 |
0.030± 0.005 |
0.026± 0.005 |
0.023± 0.006 |
0.024± 0.006 |
TOTAL |
1/1 |
0.233± 0.024 |
0.257± 0.022 |
0.248± 0.042 |
0.230± 0.029 |
Table 33. VL3. Main study. Results of blood serum biochemical examination - males
Parameter |
GROUP |
||||
0 n=10 |
1 n=10 |
2 n=10 |
3 n=10 |
||
TOTAL PROTEIN |
g/L |
60.73±2.03 |
61.53±2.11 |
61.63±2.80 |
62.01 ±3.07 |
ALBUMIN |
g/L |
32.56± 0.79 |
32.61 ± 1.02 |
32.51 ± 1.13 |
32.71 ± 1.36 |
GLOBULIN |
g/L |
28.17± 1.30 |
28.92± 1.54 |
29.12± 1.91 |
29.30± 1.88 |
A/G RATIO |
1.16± 0.03 |
1.13± 0.06 |
1.12± 0.05 |
1.12± 0.04 |
|
GLUCOSE |
mmol/L |
8.79 ± 1.78 |
8.83 ± 1.37 |
8.69 ± 0.69 |
8.88 ± 1.31 |
CHOLESTEROL |
mmol/L |
1.67 ± 0.36 |
1.58 ± 0.28 |
1.76± 0.17 |
1.83 ± 0.25 |
UREA NITROGEN |
mmol/L |
4.80 ± 0.44 |
4.69 ± 0.46 |
4.41 ± 0.53 |
5.04 ± 0.67 |
CREATININE |
pmol/L |
32.30± 3.86 |
29.90± 5.07 |
30.60±3.89 |
30.50±3.81 |
BILE ACIDS |
pmol/L |
18.30± 12.07 |
12.95± 8.86 |
15.67±9.13 |
24.76± 12.48 |
BILIRUBIN |
pmol/L |
3.91 ± 0.53 |
3.40 ± 0.24 |
3.40 ± 0.60 |
3.74 ± 0.84 |
SODIUM |
mmol/L |
144.50±2.46 |
145.50± 0.97 |
145.10± 1.29 |
146.00± 1.33 |
POTASSIUM |
mmol/L |
4.16 ± 0.28 |
4.24 ± 0.32 |
4.09 ± 0.20 |
4.21 ± 0.40 |
CHLORIDES |
mmol/L |
105.10±2.02 |
106.80± 1.40* |
107.30± 1.06* |
107.00± 1.41* |
CALCIUM |
mmol/L |
2.46 ± 0.08 |
2.48 ± 0.06 |
2.46 ± 0.05 |
2.54 ±0.05* |
x± SD;
n - number of test animals
* - statistically significant difference with p < 0.05; Dunnett's test;
Table 34.VL3. Main study. Results of blood serum biochemical examination - females
Parameter |
GROUP |
||||
0 n=12 |
1 n=12 |
2 n=12 |
3 n=11 |
||
TOTAL PROTEIN |
g/L |
56.32±4.91 |
55.75土2.91 |
57.60土4.10 |
55.53土2.82 |
ALBUMIN |
g/L |
30.77± 3.11 |
30.15土1.72 |
31.68土2.64 |
30.42土1.34 |
GLOBULIN |
g/L |
25.55± 1.96 |
25.60土1.71 |
25.92土1.65 |
25.11土1.93 |
A/G RATIO |
1.20土0.06 |
1.18土0.08 |
1.22土0.06 |
1.22土0.09 |
|
GLUCOSE |
mmol/L |
7.46土1.96 |
7.18土1.67 |
7.24土1.26 |
7.40土1.90 |
CHOLESTEROL |
mmol/L |
2.42土0.52 |
2.78土0.51 |
2.20土0.63 |
2.68土0.54 |
UREA NITROGEN |
mmol/L |
7.23 ±2.29 |
6.67土1.33 |
6.25土1.46 |
8.77土1.90 |
CREATININE |
pmol/L |
36.33± 3.20 |
33.33土2.57 |
33.08土3.60 |
40.82土5.53* |
BILE ACIDS |
pmol/L |
41.80± 32.74 |
43.12土28.28 |
39.49土25.46 |
79.07土102.36 |
BILIRUBIN |
pmol/L |
5.04 ±2.31 |
5.21土1.61 |
4.34土1.03 |
5.02土0.83 |
SODIUM |
mmol/L |
139.75± 1.96 |
139.67土2.10 |
141.50土1.62 |
140.55土1.81 |
POTASSIUM |
mmol/L |
3.94土0.71 |
4.19土0.40 |
3.96土0.22 |
4.15土0.40 |
CHLORIDES |
mmol/L |
98.92± 3.80 |
99.17土3.27 |
101.75土3.65 |
97.55土2.21 |
CALCIUM |
mmol/L |
2.48土0.06 |
2.46土0.08 |
2.49土0.10 |
2.52土0.04 |
x± SD;
n - number of test animals
* - statistically significant difference with p < 0.05; Dunnett's test
Table 35. VL3. Main study. Results of blood serum biochemical examination - (satellite groups)
Parameter |
GROUP (males) |
GROUP (females) |
|||
0 SAT n=10 |
3 SAT n=10 |
0 SAT n=10 |
3 SAT n=10 |
||
TOTAL PROTEIN |
g/L |
61.07± 1.42 |
63.10土2.10* |
0 SAT n=10 |
3 SAT n=10 |
ALBUMIN |
g/L |
31.68± 1.03 |
32.44土1.00 |
60.97± 1.40 |
61.97土2.63 |
GLOBULIN |
g/L |
29.39± 1.39 |
30.66土1.36 |
34.77± 1.29 |
35.04土1.58 |
A/G RATIO |
1.08土0.07 |
1.06土0.04 |
26.20± 0.65 |
26.93土1.52 |
|
GLUCOSE |
mmol/L |
8.29土1.27 |
7.15土0.76* |
1.33土0.06 |
1.30土0.07 |
CHOLESTEROL |
mmol/L |
2.11 ± 0.37 |
2.07土0.36 |
5.91 ± 0.94 |
5.88土0.97 |
UREA NITROGEN |
mmol/L |
5.32土0.49 |
5.35土0.46 |
1.69土0.46 |
1.53土0.39 |
CREATININE |
pmol/L |
31.90± 5.82 |
28.20土4.44 |
5.99土0.75 |
5.32土0.60* |
BILE ACIDS |
pmol/L |
21.72± 16.99 |
19.57土9.32 |
34.10±4.41 |
31.50土4.28 |
BILIRUBIN |
pmol/L |
4.10土0.54 |
3.81土0.46 |
36.94±20.24 |
27.98土18.13 |
SODIUM |
mmol/L |
144.80± 1.69 |
145.90土1.37 |
4.73土1.12 |
4.31土1.09 |
POTASSIUM |
mmol/L |
4.45土0.56 |
4.25土0.28 |
143.80± 0.79 |
145.10土0.88* |
CHLORIDES |
mmol/L |
105.10土1.79 |
105.20土1.03 |
3.92土0.35 |
3.95土0.31 |
CALCIUM |
mmol/L |
2.49土0.07 |
2.54土0.05 |
105.60土0.84 |
106.40土1.26 |
|
|
|
|
2.48土0.09 |
2.44土0.05 |
n - number of test animals
* - statistically significant difference with p < 0.05; Student's t-test;
Table 36. VL3. Main study. Results of blood serum enzymatic examination - males
Parameter |
GROUP |
|||
0 n=10 |
1 n=10 |
2 n=10 |
3 n=10 |
|
AST IU |
152.90±49.99 |
119.10±20.46 |
120.10±34.76 |
137.10±57.92 |
ALT IU |
38.70±7.86 |
37.90± 7.20 |
43.70± 14.44 |
46.50± 18.34 |
AP IU |
159.80±23.54 |
133.80± 17.78* |
133.30±24.23* |
149.30± 16.94 |
f土SD
n - number of test animals
* - statistically significant difference with p < 0.05; Dunnett's test;
Table 37. VL3. Main study. Results of blood serum enzymatic examination - females
Parameter |
GROUP |
|||
0 n=12 |
1 n=12 |
2 n=12 |
3 n=11 |
|
AST IU |
193.67±62.23 |
183.67± 81.79 |
125.75±28.44* |
179.64±57.77 |
ALT IU |
70.75± 39.81 |
86.25±47.83 |
52.00± 18.47 |
73.73±23.95 |
AP IU |
135.75±42.63 |
173.50±61.36 |
149.08±66.80 |
158.55±42.32 |
f土SD
n - number of test animals
* - statistically significant difference with p < 0.05; Dunnett's test;
Table 38. VL3. Main study. Results of blood serum enzymatic examination - (satellite groups)
Parameter |
GROUP (males) |
GROUP (females) |
|||
0 SAT n=10 |
3 SAT n=10 |
0 SAT n=10 |
3 SAT n=10 |
||
AST |
IU |
153.50± 37.19 |
107.50± 18.49* |
148.70± 33.72 |
133.00± 31.61 |
ALT |
IU |
42.40± 15.46 |
34.40± 8.75 |
29.40± 7.06 |
28.10± 7.71 |
AP |
IU |
103.30± 12.56 |
93.40± 10.94 |
51.00± 9.82 |
56.30± 15.64 |
f土SD;
n - number of test animals
* - statistically significant difference with p < 0.05; Student's t-test;
Table 39. VL3. Main study. Results of general urine examination - males
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
VOLUME mL |
14.40±6.80 |
12.00土4.95 |
16.00土9.90 |
17.40土10.33 |
SPECIFIC GRAVITY |
1.02土0.01 |
1.02土0.01 |
1.02土0.01 |
1.02土0.01 |
pH |
6.90土0.22 |
6.80土0.27 |
6.90土0.22 |
6.80土0.27 |
PROTEIN g/L |
0.30土0.41 |
0.26土0.09 |
0.18土0.16 |
0.10土0.12 |
GLUCOSE mmol/L |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
KETONE BODIES mmol/L |
0.30土0.67 |
0.10土0.22 |
0.30土0.27 |
0.20土0.27 |
BILIRUBINc |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
BLOOD Ery/pL |
0.00土0.00 |
0.00土0.00 |
2.00土4.47 |
0.00土0.00 |
UROBILINOGEN pmol/L |
3.20土0.00 |
3.20土0.00 |
3.20土0.00 |
3.20土0.00 |
LEUKOCYTES Leu/pL |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
x土SD
n - number of test animals C - qualitative test
Table 40. VL3. Main study. Results of general urine examination - females
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
VOLUME mL |
11.20± 3.90 |
14.80土3.35 |
13.60土4.34 |
15.20土3.03 |
SPECIFIC GRAVITY |
1.03土0.00 |
1.03土0.00 |
1.03土0.00 |
1.03土0.00 |
pH |
6.00土0.35 |
6.20土0.57 |
6.30土0.57 |
6.10土0.55 |
PROTEIN g/L |
0.20土0.14 |
0.04土0.05* |
0.06土0.05* |
0.04土0.05* |
GLUCOSE mmol/L |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
KETONE BODIES mmol/L |
0.20土0.27 |
0.30土0.27 |
0.20土0.27 |
0.00土0.00 |
BILIRUBINc |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
BLOOD Ery/pL |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
16.00土35.78 |
UROBILINOGEN pmol/L |
3.20土0.00 |
3.20土0.00 |
3.20土0.00 |
3.20土0.00 |
LEUKOCYTES Leu/pL |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
x ± SD;
n - number of test animals C - qualitative test
* - statistically significant difference with p < 0.05; Dunnett's test;
Table 41. VL3. Main study. Results of general urine examination - males (satellite groups)
Parameter |
GROUP (males) |
GROUP (females) |
|||
0 SAT n=5 |
3 SAT n=5 |
0 SAT n=5 |
3 SAT n=5 |
||
VOLUME |
mL |
21.60± 5.90 |
13.40±6.99 |
16.20±6.87 |
12.60土3.13 |
SPECIFIC GRAVITY |
1.02 ± 0.00 |
1.02 ±0.00 |
1.02土0.00 |
1.02土0.00 |
|
pH |
7.10 ± 0.22 |
7.10 ±0.22 |
6.80 ± 0.27 |
6.50土0.35 |
|
PROTEIN |
g/L |
0.08 ± 0.04 |
0.54 ± 0.43* |
0.00土0.00 |
0.00土0.00 |
GLUCOSE |
mmol/L |
0.00 ± 0.00 |
0.00 ±0.00 |
0.00土0.00 |
0.00土0.00 |
KETONE BODIES |
mmol/L |
1.30 ± 0.45 |
1.30 ±0.45 |
0.00土0.00 |
0.00土0.00 |
BILIRUBINc |
0.00 ± 0.00 |
0.00 ±0.00 |
0.00土0.00 |
0.00土0.00 |
|
BLOOD |
Ery/pL |
2.00 ±4.47 |
0.00 ±0.00 |
0.00土0.00 |
0.00土0.00 |
UROBILINOGEN |
pmol/L |
3.20 ± 0.00 |
3.20 ±0.00 |
3.20土0.00 |
3.20土0.00 |
LEUKOCYTES |
Leu/pL |
0.00 ± 0.00 |
6.00 ± 8.22 |
0.00土0.00 |
0.00土0.00 |
Table 42. VL3. Main study. Results of urine sediment examination - males
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
SQUAMOUS EPITHELIA |
1.20土0.45 |
1.00土0.00 |
1.40土0.55 |
1.20土0.45 |
COLUMNAR EPITHELIA |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
LEUKOCYTES |
1.00土0.00 |
1.00土0.00 |
1.00土0.00 |
1.00土0.00 |
ERYTHROCYTES |
1.20土0.45 |
1.20土0.45 |
1.20土0.45 |
1.40土0.55 |
BACTERIA |
3.60土0.55 |
4.00土0.71 |
4.20土0.84 |
3.80土0.45 |
CRYSTALS |
1.80土1.10 |
1.40土1.34 |
1.20土1.10 |
2.00土1.22 |
f± SD;
n - number of test animals
Scale of quantitative evaluation of urine sediment elements in single animals in group: 1-single/slide, 2 - single/HPF (HPF-per High Power Field), 3 - few/HPF, 4 - quite numerous/HPF, 5 - numerous/HPF, 6-very numerous/HPF, 7-plantifull/HPF
Table 43. VL3. Main study. Results of urine sediment examination - females
Parameter |
GROUP |
|||
0 n=5 |
1 n=5 |
2 n=5 |
3 n=5 |
|
SQUAMOUS EPITHELIA |
1.00土0.00 |
1.00土0.00 |
1.00土0.00 |
1.00土0.00 |
COLUMNAR EPITHELIA |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
0.00土0.00 |
LEUKOCYTES |
1.00土0.00 |
1.00土0.00 |
1.00土0.00 |
1.00土0.00 |
ERYTHROCYTES |
1.20土0.45 |
1.00土0.00 |
1.00土0.00 |
1.20土0.45 |
BACTERIA |
2.60土1.34 |
2.60土0.89 |
3.00土0.71 |
2.80土0.84 |
CRYSTALS |
0.60 ± 1.34 |
1.60土1.67 |
1.80土1.79 |
1.60土1.52 |
x± SD;
n - number of test animals
Scale of quantitative evaluation of urine sediment elements in single animals in group:
1-single/slide, 2 - single/HPF (HPF-per High Power Field), 3 - few/HPF, 4 - quite numerous/HPF, 5 - numerous/HPF, 6-very numerous/HPF, 7-plantifull/HPF
Table 44. VL3. Main study. Results of urine sediment examination - (satellite groups)
Parameter |
GROUP (males) |
GROUP (females) |
||
0 SAT n=5 |
3 SAT n=5 |
0 SAT n=5 |
3 SAT n=5 |
|
SQUAMOUS EPITHELIA |
1.20土0.45 |
1.20土0.45 |
1.00 ± 0.00 |
1.00 ±0.00 |
COLUMNAR EPITHELIA |
0.00土0.00 |
0.00土0.00 |
0.00 ± 0.00 |
0.00 ±0.00 |
LEUKOCYTES |
1.00土0.00 |
1.00土0.00 |
1.00 ± 0.00 |
1.00 ±0.00 |
ERYTHROCYTES |
1.20土0.45 |
1.20土0.45 |
1.20 ± 0.45 |
1.20 ±0.45 |
BACTERIA |
4.20土0.84 |
2.40土0.89* |
2.80 ± 1.30 |
3.40 ± 1.14 |
CRYSTALS |
0.60土1.34 |
2.60土0.89 |
1.00 ±2.24 |
0.00 ±0.00 |
x± SD;
n - number of test animals
Scale of quantitative evaluation of urine sediment elements in single animals in group:
1-single/slide, 2 - single/HPF (HPF-per High Power Field), 3 - few/HPF, 4 - quite numerous/HPF, 5 - numerous/HPF, 6-very numerous/HPF, 7-plantifull/HPF
Table 45. VL3. Main study. Results of hormonal examination - adult males
Parameter |
GROUP |
|||
0 n=10 |
1 n=10 |
2 n=10 |
3 n=10 |
|
TT4 |
4.88 ±0.48 |
5.13 ± 0.63 |
4.91 ± 0.43 |
4.94 ± 0.64 |
Table 46. VL3. Main study. Results of hormonal examination - adult males (satellite groups)
|
GROUP |
|
Parameter |
0 SAT |
3 SAT |
|
n=10 |
n=10 |
TT4 |
4.98 ± 0.64 |
5.56 ±0.77 |
Table 47. VL3. Main study. Gross lesions: euthanized animals
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||||||
Examined organ |
Type of change |
0 |
1 |
2 |
3 |
||||
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
||
Trachea |
filled with foamy liquid |
|
2 [14,21] |
|
2 [17,18] |
|
1 [13] |
|
|
|
ecchymosis of 1/3 organ area |
_ |
_ |
_ |
1 [17] |
_ |
_ |
_ |
_ |
Thymus |
petechiae |
_ |
1 [17] |
_ |
1 [18] |
_ |
1 [16] |
_ |
1 [22] |
|
decreased |
_ |
1 [17] |
- |
2 [16,17] |
_ |
1 [13] |
_ |
_ |
|
increased |
2 [6,10] |
1 [17] |
3 [3,6,9] |
5 [11,14,16,18, 22] |
- |
2 [13,17] |
2 [6,10] |
1 [11] |
|
bright |
_ |
1 [20] |
_ |
1 [18] |
_ |
1 [16] |
_ |
1 [22] |
Lungs |
bright foci above the surface |
_ |
3 [11,19,20] |
_ |
5 [12,16,17, 18,19] |
_ |
1 [20] |
_ |
1 [22] |
|
foci collapsed under the surface |
_ |
_ |
_ |
1 [20] |
_ |
_ |
_ |
_ |
|
red |
_ |
_ |
_ |
1 [22] |
_ |
_ |
_ |
_ |
Table 47 cont. VL3. Main study.Gross lesions: euthanized animals
Examined organ |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||||||
0 |
1 |
2 |
3 |
||||||
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
||
Lungs |
marbled |
2 [2,10] |
3 [14,18,21] |
_ |
2 [16,21] |
_ |
5 [13,15,19,21, 22] |
1 [10] |
4 [11,13,14, 18] |
dark red |
1 [6] |
_ |
3 [3,6,9] |
- |
- |
- |
_ |
1 [17] |
|
dark red foci |
1 [2] |
2 [17,20] |
_ |
1 [18] |
_ |
1 [16] |
_ |
_ |
|
dark red area |
1 [8] |
6 [12,14,15,16, 21,22] |
1 m |
2 [15,20] |
2 [3,4] |
2 [14,17] |
_ |
4 [11,12,14,15] |
|
Heart |
slightly increased |
1 [10] |
_ |
_ |
1 [18] |
_ |
1 [18] |
_ |
_ |
Cecum |
congestion |
_ |
_ |
- |
1 [15] |
- |
- |
_ |
_ |
Ovaries |
congestion |
_ |
_ |
- |
- |
- |
1 [16] |
_ |
_ |
[]-computer numbers of animals with gross changes n - number of tested animals
Table 47 cont. VL3. Main study.Gross lesions: euthanized animals
Examined organ |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||||||
0 |
1 |
2 |
3 |
||||||
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
||
Lungs |
marbled |
2 [2,10] |
3 [14,18,21] |
_ |
2 [16,21] |
_ |
5 [13,15,19,21, 22] |
1 [10] |
4 [11,13,14, 18] |
dark red |
1 [6] |
_ |
3 [3,6,9] |
- |
- |
- |
_ |
1 [17] |
|
dark red foci |
1 [2] |
2 [17,20] |
_ |
1 [18] |
_ |
1 [16] |
_ |
_ |
|
dark red area |
1 [8] |
6 [12,14,15,16, 21,22] |
1 m |
2 [15,20] |
2 [3,4] |
2 [14,17] |
_ |
4 [11,12,14,15] |
|
Heart |
slightly increased |
1 [10] |
_ |
_ |
1 [18] |
_ |
1 [18] |
_ |
_ |
Cecum |
congestion |
_ |
_ |
- |
1 [15] |
- |
- |
_ |
_ |
Ovaries |
congestion |
_ |
_ |
- |
- |
- |
1 [16] |
_ |
_ |
Table 47 cont. VL3. Main study.Gross lesions: euthanized animals
Examined organ |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||||||
0 |
1 |
2 |
3 |
||||||
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
males n=10 |
females n=12 |
||
Uterus |
congestion |
_ |
1 [17] |
- |
1 [16] |
- |
2 [13,16] |
_ |
1 [22] |
partially cyanosis |
_ |
_ |
_ |
_ |
_ |
1 [16] |
_ |
_ |
|
expanded |
_ |
- |
_ |
_ |
_ |
1 [16] |
_ |
1 [22] |
|
thickened wall |
_ |
_ |
_ |
_ |
_ |
_ |
_ |
1 [22] |
|
filled with clear liquid |
_ |
_ |
_ |
_ |
_ |
_ |
_ |
1 [22] |
|
cyanosis of the horns |
_ |
_ |
- |
1 [12] |
- |
3 [19,20,21] |
_ |
_ |
|
Small intestine |
congestion |
_ |
_ |
_ |
1 [15] |
_ |
_ |
_ |
_ |
Table 48.VL3. Main study. Gross lesions: euthanized animals (satellite groups)
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
|
|
0 SAT |
3 SAT |
||
Examined organ |
Type of change |
males n=10 |
females n=10 |
males n=10 |
females n=10 |
Trachea |
filled with foamy liquid |
1 [31] |
1 [48] |
3 [32,34,40] |
1 [47] |
Thymus |
petechiae |
3 [31,33,35] |
5 [41,45,48,49,50] |
5 [32,34,35,36,37] |
4 [41,43,48,49] |
decreased |
1 [37] |
1 [48] |
1 [37] |
_ |
|
|
bright red |
4 [31,33,34,39] |
1 [43] |
3 [33,35,38] |
_ |
|
bright red foci |
1 [33] |
1 [48] |
_ |
_ |
|
bright red foci above the surface |
2 [31,39] |
2 [43,50] |
3 [33,35,39] |
2 [41,48] |
Lungs |
dark red foci collapsed under the surface |
_ |
_ |
1 [33] |
_ |
|
increased |
2 [32,36] |
1 [48] |
2 [32,34] |
1 [47] |
|
marbled |
6 [32,35,36,37,39, 40] |
1 [46] |
7 [31,32,33,34,37, 39,40] |
1 [47] |
Table 48 cont. VL3. Main study.Gross lesions: euthanized animals (satellite groups)
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
|
|
0 SAT |
3 SAT |
||
Examined organ |
Type of change |
males n=10 |
females n=10 |
males n=10 |
females n=10 |
|
dark red |
_ |
_ |
1 [36] |
一 |
Lungs |
dark red foci |
3 [31,33,34] |
6 [44,45,47,48,49, 50] |
1 [38] |
8 [41,42,43,44,45, 46,48,49] |
|
dark red foci under the surface |
_ |
_ |
1 [39] |
一 |
|
doughy consistency |
_ |
_ |
一 |
1 [50] |
Heart |
increased |
4 [35,36,37,40] |
_ |
2 [34,37] |
一 |
congestion |
_ |
_ |
2 [36,38] |
一 |
|
Liver |
lesion- 7mm long, yellowish, grainy, on the visceral side of the left lobe |
_ |
1 [46] |
一 |
一 |
Spleen |
increased |
1 [35] |
一 |
一 |
一 |
Cecum |
congestion |
1 [36] |
_ |
一 |
一 |
Table 48 cont. VL3. Main study.Gross lesions: euthanized animals (satellite groups)
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
Examined organ |
Type of change |
0 SAT |
3 SAT |
||
males |
females |
males |
females |
||
|
|
n=10 |
n=10 |
n=10 |
n=10 |
Colon |
petechiae |
1 [35] |
一 |
一 |
一 |
Mandibular lymph nodes |
petechiae |
_ |
一 |
1 [34] |
2 [41,43] |
Kidneys |
increased |
2 [35,40] |
一 |
一 |
一 |
Ovaries |
dark red foci |
_ |
1 [45] |
一 |
1 [48] |
|
congestion |
_ |
1 [43] |
一 |
1 [48] |
Uterus |
slightly expanded |
一 |
4 [43,44,48,50] |
一 |
2 [47,49] |
|
filled with |
|
4 |
|
2 |
|
clear liquid |
_ |
[43,44,48,50] |
一 |
[47,49] |
Table 48 cont. VL3. Main study.Gross lesions: euthanized animals (satellite groups)
Examined organ |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||
0 SAT |
3SAT |
||||
males n=10 |
females n=10 |
males n=10 |
females n=10 |
||
lesion- yellowish,elastic, size 5x2mm in subcutaneous tissue of cervical area above the salivary gland |
_ |
_ |
1 [31] |
_ |
|
lesion - yellowish, 5 mm long located in fat tissue in left ovary area |
_ |
_ |
_ |
1 [41] |
|
lesion - yellowish, 5 mm long located in fat tissue in pelvis cavity inlet area |
_ |
_ |
_ |
1 [46] |
|
Cranial cavity filled with blood |
_ |
_ |
1 [37] |
_ |
|
Table 49. VL3. Main study. Absolute weights of internal organs [ mg ] - males.
|
Group |
|||||||||||
Examined organ |
0 n=10 |
1 n=10 |
2 n=10 |
3 n=10 |
||||||||
Brain |
2047.400 |
土 |
85.481 |
1999.700 |
土 |
134.953 |
2096.300 |
土 |
80.760 |
2073.200 |
土 |
73.295 |
Pituitary gland |
11.400 |
土 |
1.838 |
10.600 |
土 |
1.578 |
10.800 |
土 |
1.874 |
8.800 |
土 |
1.751* |
Thyroid |
24.800 |
土 |
5.770 |
23.700 |
土 |
3.683 |
25.800 |
土 |
3.084 |
19.900 |
土 |
4.999 |
Thymus |
327.000 |
土 |
50.859 |
299.500 |
土 |
45.887 |
283.800 |
土 |
40.881 |
268.500 |
土 |
66.304 |
Heart |
1124.200 |
土 |
302.274 |
1012.500 |
土 |
124.886 |
1064.500 |
土 |
96.709 |
980.300 |
土 |
97.803 |
Liver |
11540.600 |
土 |
1760.413 |
11315.800 |
土 |
970.273 |
11666.100 |
土 |
2188.889 |
11380.600 |
土 |
1101.959 |
Spleen |
731.100 |
土 |
94.090 |
647.100 |
土 |
77.229 |
655.900 |
土 |
66.534 |
611.400 |
土 |
84.733* |
Kidneys |
2606.200 |
土 |
250.519 |
2605.000 |
土 |
213.370 |
2591.100 |
土 |
339.513 |
2633.500 |
土 |
200.172 |
Adrenals |
83.700 |
土 |
10.636 |
79.700 |
土 |
10.489 |
78.600 |
土 |
14.416 |
75.800 |
土 |
12.515 |
Testicles |
3655.600 |
土 |
179.424 |
3575.400 |
土 |
226.663 |
3590.500 |
土 |
285.645 |
3677.700 |
土 |
233.694 |
Epidydimides |
1415.400 |
土 |
115.627 |
1386.800 |
土 |
117.212 |
1387.400 |
土 |
154.952 |
1414.200 |
土 |
187.585 |
Prostate with vesicular and coagulating glands |
2143.800 |
土 |
260.990 |
2244.000 |
土 |
235.569 |
2290.300 |
土 |
195.046 |
2363.500 |
土 |
408.443 |
Levator ani |
310.000 |
土 |
19.299 |
308.800 |
土 |
41.886 |
302.000 |
土 |
31.337 |
280.900 |
土 |
38.240 |
Bulbocavernosus muscle complex |
784.900 |
土 |
80.622 |
767.500 |
土 |
97.903 |
792.100 |
土 |
72.301 |
810.200 |
土 |
93.866 |
Cowper’s glans |
108.300 |
土 |
19.482 |
101.200 |
土 |
14.038 |
105.800 |
土 |
17.492 |
112.600 |
土 |
15.291 |
Glans penis |
90.900 |
土 |
14.708 |
92.400 |
土 |
8.140 |
99.700 |
土 |
13.200 |
95.300 |
土 |
12.996 |
Y ± SD
* - statistically significant difference at p < 0.05 - Dunnett's test n - number of tested animals
Table 50. VL3. Main study. Absolute weights of internal organs [ mg ] - females.
|
Group |
|||||||||||
Examined organ |
0 n=12 |
1 n=12 |
2 n=12 |
3 n=12 |
||||||||
|
||||||||||||
Brain |
1898.583 |
土 |
80.4357 |
1905.250 |
土 |
72.227 |
1909.667 |
土 |
72.089 |
1896.417 |
土 |
114.773 |
Pituitary gland |
15.417 |
土 |
1.831955 |
14.833 |
土 |
1.946 |
13.250 |
土 |
1.485* |
13.417 |
土 |
1.443* |
Thyroid |
17.917 |
|
4.521833 |
18.167 |
|
2.517 |
17.417 |
|
3.397 |
18.500 |
|
3.090 |
Thymus |
240.750 |
土 |
101.7771 |
184.917 |
土 |
53.294 |
220.333 |
土 |
71.970 |
211.500 |
土 |
80.636 |
Heart |
795.000 |
土 |
92.63222 |
813.417 |
土 |
89.118 |
793.750 |
土 |
65.840 |
841.417 |
土 |
89.465 |
Liver |
10809.250 |
土 |
1819.769 |
11455.500 |
土 |
1494.417 |
10139.333 |
土 |
1436.985 |
11069.250 |
土 |
1751.459 |
Spleen |
615.250 |
土 |
52.42505 |
621.417 |
土 |
95.102 |
631.250 |
土 |
80.462 |
622.750 |
土 |
108.084 |
Kidneys |
1868.667 |
土 |
203.5826 |
1885.083 |
土 |
162.555 |
1867.667 |
土 |
133.189 |
1847.583 |
土 |
190.692 |
Adrenals |
94.833 |
土 |
10.18763 |
94.000 |
土 |
13.253 |
91.667 |
土 |
11.781 |
97.917 |
土 |
9.605 |
Ovaries |
125.417 |
土 |
30.81457 |
116.667 |
土 |
16.472 |
126.417 |
土 |
33.489 |
123.167 |
土 |
14.721 |
Uterus with cervix |
547.750 |
土 |
215.9192 |
454.833 |
土 |
100.221 |
537.167 |
土 |
173.530 |
531.250 |
土 |
206.144 |
f ± SD
* - statistically significant difference at p < 0.05 - Dunnett's test n - number of tested animals
Table 51. VL3. Main study. Relative weights of internal organs [%] - males.
|
Group |
|||||||||||
Examined organ |
|
0 |
|
|
1 |
|
|
2 |
|
|
3 |
|
|
|
n=10 |
|
|
n=10 |
|
|
n=10 |
|
|
n=10 |
|
Brain |
0.496 |
土 |
0.043 |
0.491 |
土 |
0.054 |
0.511 |
土 |
0.049 |
0.514 |
土 |
0.046 |
Pituitary gland |
0.0006 |
土 |
0.0001 |
0.0005 |
土 |
0.0001 |
0.0005 |
土 |
0.0001 |
0.0004 |
土 |
0.0001* |
Thyroid |
0.006 |
土 |
0.001 |
0.006 |
土 |
0.001 |
0.006 |
土 |
0.001 |
0.005 |
土 |
0.001* |
Thymus |
0.079 |
土 |
0.009 |
0.073 |
土 |
0.011 |
0.069 |
土 |
0.010 |
0.066 |
土 |
0.016 |
Heart |
0.271 |
土 |
0.073 |
0.247 |
土 |
0.016 |
0.258 |
土 |
0.014 |
0.242 |
土 |
0.013 |
Liver |
2.765 |
土 |
0.216 |
2.770 |
土 |
0.225 |
2.811 |
土 |
0.326 |
2.806 |
土 |
0.166 |
Spleen |
0.176 |
土 |
0.014 |
0.158 |
土 |
0.015* |
0.159 |
土 |
0.016* |
0.150 |
土 |
0.014* |
Kidneys |
0.629 |
土 |
0.062 |
0.638 |
土 |
0.051 |
0.627 |
土 |
0.056 |
0.650 |
土 |
0.030 |
Adrenals |
0.020 |
土 |
0.002 |
0.020 |
土 |
0.003 |
0.019 |
土 |
0.003 |
0.019 |
土 |
0.002 |
Testicles |
0.884 |
土 |
0.070 |
0.877 |
土 |
0.079 |
0.873 |
土 |
0.088 |
0.910 |
土 |
0.074 |
Epidydimides |
0.342 |
土 |
0.026 |
0.340 |
土 |
0.028 |
0.336 |
土 |
0.024 |
0.349 |
土 |
0.037 |
Prostate with vesicular and |
0.518 |
土 |
0.063 |
0.551 |
土 |
0.069 |
0.556 |
土 |
0.048 |
0.582 |
土 |
0.088 |
coagulating glands |
|
|
|
|
|
|
|
|
|
|
|
|
Levator ani |
0.075 |
土 |
0.010 |
0.075 |
土 |
0.009 |
0.073 |
土 |
0.008 |
0.069 |
土 |
0.006 |
Bulbocavernosus muscle complex |
0.190 |
土 |
0.020 |
0.189 |
土 |
0.032 |
0.193 |
土 |
0.020 |
0.200 |
土 |
0.014 |
Cowper’s glans |
0.026 |
土 |
0.005 |
0.025 |
土 |
0.003 |
0.026 |
土 |
0.004 |
0.028 |
土 |
0.003 |
Glans penis |
0.022 |
土 |
0.005 |
0.023 |
土 |
0.003 |
0.024 |
土 |
0.003 |
0.023 |
土 |
0.002 |
f ± SD
* - statistically significant difference at p < 0.05 - Dunnett's test n - number of tested animals
Table 52. VL3. Main study. Relative weights of internal organs [%] - females.
Examined organ |
Group |
|||||||||||
0 n=12 |
1 n=12 |
2 n=12 |
3 n=12 |
|||||||||
Brain |
0.672 |
土 |
0.031 |
0.662 |
土 |
0.029 |
0.688 |
土 |
0.049 |
0.668 |
土 |
0.050 |
Pituitary gland |
0.005 |
土 |
0.001 |
0.005 |
土 |
0.001 |
0.005 |
土 |
0.001 |
0.005 |
土 |
0.001 |
Thyroid |
0.006 |
土 |
0.001 |
0.006 |
土 |
0.001 |
0.006 |
土 |
0.001 |
0.007 |
土 |
0.001 |
Thymus |
0.085 |
土 |
0.034 |
0.064 |
土 |
0.018 |
0.079 |
土 |
0.025 |
0.076 |
土 |
0.035 |
Heart |
0.281 |
土 |
0.025 |
0.282 |
土 |
0.023 |
0.285 |
土 |
0.012 |
0.296 |
土 |
0.030 |
Liver |
3.819 |
土 |
0.615 |
3.968 |
土 |
0.439 |
3.641 |
土 |
0.461 |
3.873 |
土 |
0.444 |
Spleen |
0.218 |
土 |
0.017 |
0.215 |
土 |
0.030 |
0.227 |
土 |
0.027 |
0.218 |
土 |
0.029 |
Kidneys |
0.660 |
土 |
0.055 |
0.654 |
土 |
0.048 |
0.673 |
土 |
0.061 |
0.650 |
土 |
0.060 |
Adrenals |
0.034 |
土 |
0.003 |
0.033 |
土 |
0.004 |
0.033 |
土 |
0.005 |
0.034 |
土 |
0.004 |
Ovaries |
0.044 |
土 |
0.012 |
0.041 |
土 |
0.006 |
0.046 |
土 |
0.012 |
0.043 |
土 |
0.005 |
Uterus with cervix |
0.193 |
土 |
0.071 |
0.158 |
土 |
0.037 |
0.194 |
土 |
0.065 |
0.190 |
土 |
0.090 |
Table 53. VL3. Main study. Absolute weights of internal organs [mg] - males (satellite groups).
|
Group |
|||||
Examined organ |
|
0 SAT |
|
|
3SAT |
|
|
|
n=10 |
|
|
n=10 |
|
Brain |
2020.600 |
土 |
108.063 |
2004.400 |
土 |
137.999 |
Pituitary gland |
9.300 |
土 |
1.494 |
9.900 |
土 |
1.969 |
Thyroid |
24.300 |
土 |
3.368 |
23.900 |
土 |
3.784 |
Thymus |
295.400 |
土 |
36.692 |
340.100 |
土 |
71.191 |
Heart |
1046.300 |
土 |
159.712 |
1096.100 |
土 |
164.658 |
Liver |
12122.900 |
土 |
2528.654 |
12430.000 |
土 |
1738.979 |
Spleen |
737.200 |
土 |
157.512 |
787.500 |
土 |
133.114 |
Kidneys |
2720.000 |
土 |
285.898 |
2842.400 |
土 |
236.942 |
Adrenals |
77.400 |
土 |
12.158 |
76.400 |
土 |
9.947 |
Testicles |
3576.700 |
土 |
245.092 |
3498.200 |
土 |
238.550 |
Epidydimides |
1493.700 |
土 |
116.091 |
1446.300 |
土 |
131.374 |
Prostate with vesicular and coagulating glands |
2407.000 |
土 |
307.769 |
2434.000 |
土 |
340.063 |
Levator ani |
303.000 |
土 |
49.169 |
294.900 |
土 |
32.959 |
Bulbocavernosus muscle complex |
767.200 |
土 |
106.542 |
787.300 |
土 |
75.918 |
Cowper’s glans |
112.300 |
土 |
19.097 |
112.500 |
土 |
30.064 |
Glans penis |
89.300 |
土 |
15.492 |
88.100 |
土 |
13.420 |
Table 54 VL3. Main study. Absolute weights of internal organs [mg] - females (satellite groups).
Examined organ |
Group |
|||||
0 SAT n=10 |
3SAT n=10 |
|||||
Brain |
1935.700 |
土 |
110.800 |
1943.300 |
土 |
88.000 |
Pituitary gland |
13.900 |
土 |
4.458 |
13.000 |
土 |
2.055 |
Thyroid |
12.700 |
土 |
3.129 |
12.800 |
土 |
3.615 |
Thymus |
299.100 |
土 |
125.152 |
334.700 |
土 |
87.987 |
Heart |
658.800 |
土 |
67.952 |
678.900 |
土 |
56.011 |
Liver |
6631.700 |
土 |
657.827 |
6540.500 |
土 |
824.436 |
Spleen |
497.800 |
土 |
23.976 |
512.800 |
土 |
56.233 |
Kidneys |
1648.500 |
土 |
160.506 |
1745.900 |
土 |
194.600 |
Adrenals |
81.200 |
土 |
7.885 |
87.900 |
土 |
8.736 |
Ovaries |
162.200 |
土 |
21.175 |
173.800 |
土 |
24.275 |
Uterus with cervix |
704.300 |
土 |
293.645 |
733.100 |
土 |
149.730 |
Table 55.VL3. Main study. Relative weights of internal organs [%] - males (satellite groups).
|
Group |
|||||
Examined organ |
|
0 SAT |
|
|
3SAT |
|
|
|
n=10 |
|
|
n=10 |
|
Brain |
0.478 |
土 |
0.064 |
0.461 |
土 |
0.047 |
Pituitary gland |
0.000 |
土 |
0.0001 |
0.000 |
土 |
0.000 |
Thyroid |
0.006 |
土 |
0.001 |
0.005 |
土 |
0.001 |
Thymus |
0.070 |
土 |
0.011 |
0.078 |
土 |
0.017 |
Heart |
0.244 |
土 |
0.022 |
0.250 |
土 |
0.026 |
Liver |
2.812 |
土 |
0.342 |
2.833 |
土 |
0.241 |
Spleen |
0.171 |
土 |
0.021 |
0.180 |
土 |
0.026 |
Kidneys |
0.636 |
土 |
0.025 |
0.650 |
土 |
0.032 |
Adrenals |
0.018 |
土 |
0.003 |
0.017 |
土 |
0.002 |
Testicles |
0.842 |
土 |
0.085 |
0.801 |
土 |
0.043 |
Epidydimides |
0.351 |
土 |
0.030 |
0.331 |
土 |
0.027 |
Prostate with vesicular and coagulating glands |
0.567 |
土 |
0.087 |
0.556 |
土 |
0.069 |
Levator ani |
0.071 |
土 |
0.008 |
0.068 |
土 |
0.007 |
Bulbocavernosus muscle complex |
0.180 |
土 |
0.022 |
0.181 |
土 |
0.021 |
Cowper’s glans |
0.026 |
土 |
0.005 |
0.026 |
土 |
0.007 |
Glans penis |
0.021 |
土 |
0.005 |
0.020 |
土 |
0.003 |
Table 56.VL3. Main study. Relative weights of internal organs [%] - females (satellite groups).
Examined organ |
Group |
|||||
0 SAT n=10 |
3SAT n=10 |
|||||
Brain |
0.745 |
土 |
0.048 |
0.751 |
土 |
0.051 |
Pituitary gland |
0.005 |
土 |
0.002 |
0.005 |
土 |
0.001 |
Thyroid |
0.005 |
土 |
0.001 |
0.005 |
土 |
0.001 |
Thymus |
0.114 |
土 |
0.043 |
0.128 |
土 |
0.032 |
Heart |
0.253 |
土 |
0.017 |
0.261 |
土 |
0.011 |
Liver |
2.548 |
土 |
0.188 |
2.509 |
土 |
0.162 |
Spleen |
0.192 |
土 |
0.016 |
0.198 |
土 |
0.023 |
Kidneys |
0.633 |
土 |
0.038 |
0.671 |
土 |
0.042* |
Adrenals |
0.031 |
土 |
0.003 |
0.034 |
土 |
0.003 |
Ovaries |
0.062 |
土 |
0.007 |
0.067 |
土 |
0.010 |
Uterus with cervix |
0.269 |
土 |
0.108 |
0.285 |
土 |
0.066 |
* - statistically significant difference at p < 0.05 - student's T-test n - number of tested animals
Table 57. VL3. Main study. Histopathological lesions: group 0, group 1,group 2, group 3.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||||||
Examined |
Type of change |
0f |
1m |
2m |
3f |
||||
organ / tissue |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
|
Pituitary |
microcyst |
2 [1,4] |
- |
- |
- |
- |
- |
- |
一 |
Thyroid |
congestion |
- |
- |
- |
- |
- |
- |
1 [9] |
一 |
parathyroid hypertrophy |
- |
1 P2] |
- |
- |
- |
- |
- |
一 |
|
|
congestion |
1 [2] |
- |
- |
1 [18] |
- |
- |
2 [3,9] |
一 |
Thymus |
erythrocytorrhagia |
- |
- |
- |
- |
- |
- |
1 [3] |
一 |
effusion |
- |
1 [22] |
- |
1 [17] |
|
|
- |
一 |
|
|
lymphocytic tissue depletion |
- |
1 [17] |
- |
- |
- |
1 [13] |
- |
一 |
|
erythrocytorrhagia |
3 [1,2,10] |
- |
- |
1 [18] |
- |
- |
1 [8] |
一 |
|
leukocyte infiltration |
1 [10] |
- |
- |
|
- |
- |
1 [10] |
|
Heart |
foci of fibrosis |
1 [10] |
- |
- |
- |
- |
- |
- |
一 |
|
foci of fatty degeneration |
1 [10] |
- |
- |
|
- |
- |
- |
一 |
|
left ventricular myocardium hypertrophy |
- |
- |
- |
1 [18] |
- |
1 [18] |
- |
一 |
Table 57 cont. VL3. Main study. Histopathological lesions: group 0, group 1,group 2, group 3.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||||||
Examined |
Type of change |
0f |
1m |
2m |
3f |
||||
organ / tissue |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
|
Heart |
left and right ventricular myocardium hypertrophy |
1 [10] |
- |
- |
- |
- |
- |
- |
一 |
|
congestion |
1 [6] |
- |
3 [3,6,9] |
2 [19,20] |
1 [2] |
2 [14,22] |
2 [1,5] |
2 [11,13] |
|
erythrocytorrhagia |
5 P，6,8,9,10] |
9 [12,13,14, 15,16,18, 20,21,22] |
4 [3,6,7,9] |
10 [11,14,15, 16,17,18, 19,20,21, 22] |
3 P,3,4] |
9 [13,14,15, 16,17,19, 20,21,22] |
6 [1,2,4,5,7, 10] |
8 [11,12,13, 14,15,17, 18,22] |
|
edema |
3 P，6,10] |
4 [13,14,18, 21] |
3 [3,6,9] |
2 [11,17] |
- |
2 [17,22] |
2 [5,10] |
1 [18] |
Lungs |
emphysema |
4 [2,4,8,10] |
9 [11,13,14, 15,17,19, 20,21,22] |
- |
10 [11,12,14, 15,16,18, 19,20,21, 22] |
1 [2] |
3 [13,14,21] |
5 [1,4,5,6,10] |
3 [11,18,22] |
|
foamy cell infiltration |
2 [7,8] |
7 [11,15,17, 18,20,21, 22] |
- |
3 [11,16,17] |
- |
4 [13,19,20, 21] |
1 [6] |
1 [18] |
|
lymphocyte infiltration |
1 [8] |
- |
- |
- |
- |
- |
1 [10] |
一 |
|
leukocyte infiltration |
1 [10] |
- |
- |
- |
- |
- |
- |
一 |
Table 57 cont. VL3. Main study. Histopathological lesions: group 0, group 1,group 2, group 3.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||||||
Examined |
Type of change |
0f |
1m |
2m |
3f |
||||
organ / tissue |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
|
|
congestion |
1 [10] |
2 [15,17] |
- |
- |
- |
- |
1 [9] |
一 |
|
effusion |
1 [10] |
- |
- |
- |
- |
- |
- |
一 |
Liver |
bile ducts proliferation |
1 [10] |
- |
- |
- |
- |
- |
- |
一 |
|
Browicz-Kupffer cell proliferation |
1 [10] |
- |
- |
- |
- |
- |
- |
一 |
|
hepatocyte fine- droplet fatty degeneration |
1 [3] |
- |
- |
- |
- |
- |
- |
一 |
Spleen |
congestion |
- |
1 [18] |
- |
- |
- |
- |
- |
一 |
depletion of the white pulp |
- |
2 [17,18] |
- |
- |
- |
- |
- |
一 |
|
Pancreas |
islets of Langerhans hypertrophy |
1 |
1 P1] |
- |
- |
- |
- |
3 [4,6,8] |
一 |
ducts proliferation |
- |
1 P1] |
- |
- |
- |
- |
- |
一 |
|
Jejunum |
lymphatic follicle reactive hyperplasia |
1 [5] |
- |
- |
- |
- |
- |
- |
一 |
Ileum |
lymphocyte infiltration |
- |
1 [11] |
- |
- |
- |
- |
- |
一 |
Table 57 cont. VL3. Main study. Histopathological lesions: group 0, group 1,group 2, group 3.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||||||
Examined |
Type of change |
0f |
1m |
2m |
3f |
||||
organ / tissue |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
|
Cecum |
lymphatic follicle reactive hyperplasia |
1 [7] |
- |
- |
- |
- |
- |
- |
1 [14] |
Colon |
lymphatic follicle reactive hyperplasia |
1 [4] |
- |
- |
- |
- |
- |
2 P,4] |
1 [14] |
|
erythrocytorrhagia |
1 [8] |
1 [15] |
- |
- |
- |
- |
1 [2] |
一 |
|
glomeruli congestion |
6 [1,2,3,5a 7] |
4 [12,17,20, 22] |
- |
- |
- |
- |
10 [1,2,3,4,5,6,7 8,9,10] |
6 [13,14,15, 17,20,22] |
|
lymphocyte infiltration |
- |
2 [12,21] |
- |
- |
- |
- |
1 [8] |
一 |
Kidneys |
glomeruli hypertrophy |
9 [1,2,3,5,6,7,8 ,9,10] |
7 [12,15,17, 19,20,21, 22] |
- |
- |
- |
- |
9 [1,3,4,5,6,7,8 9,10] |
9 [12,13,14, 15,17,19, 20,21,22] |
glomeruli atrophy |
4 [1,8,9,10] |
5 [15,17,18, 20,21] |
- |
- |
- |
- |
10 [1,2,3,4,5,6,7 8,9,10] |
8 [12,13,14, 15,19,20, 21,22] |
|
|
Bowman's capsule extension |
1 [9] |
2 [18,21] |
- |
- |
- |
- |
6 [2,3,5,6,8,9] |
一 |
|
retention microcysts |
- |
2 [12,15] |
- |
- |
- |
- |
1 [8] |
2 [14,21] |
|
hyaline casts |
- |
1 [12] |
- |
- |
- |
- |
- |
一 |
Table 57 cont. VL3. Main study. Histopathological lesions: group 0, group 1,group 2, group 3.
Examined organ / tissue |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||||||
0f |
1m |
2m |
3f |
||||||
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
||
Adrenals |
left adrenal gland atrophy with medulla microcyst; right adrenal gland hyperplasia |
- |
- |
- |
- |
- |
- |
- |
1 [11] |
Urinary bladder |
lymphocyte infiltration |
- |
- |
- |
- |
- |
- |
5 [1,3,5,6,7] |
一 |
Prostate |
congestion |
2 [1,9] |
|
- |
- |
- |
- |
2 [2,8] |
|
edema |
2 [1,9] |
- |
- |
- |
- |
- |
- |
一 |
|
lymphocyte infiltration |
5 [1,3,7,9,10] |
- |
3 P,4,9] |
- |
6 [2,4,5,7,8,9] |
- |
6 [2,3,5,8,9, 10] |
一 |
|
Uterus |
congestion |
- |
- |
- |
1 [16] |
- |
2 [16,20] |
- |
一 |
effusion |
- |
- |
- |
2 [12,16] |
- |
2 [20,21] |
- |
一 |
|
fibroid |
- |
- |
- |
2 [12,16] |
- |
1 [20] |
- |
一 |
|
Mandibular lymph nodes |
congestion |
1 [6] |
- |
- |
- |
- |
- |
- |
一 |
hyperplasia |
1 [6] |
- |
- |
- |
- |
- |
- |
一 |
|
lymphocytic tissue depletion |
- |
1 [17] |
- |
- |
- |
- |
- |
一 |
|
Table 57 cont. VL3. Main study. Histopathological lesions: group 0, group 1,group 2, group 3.
Examined organ / tissue |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||||||
0f |
1m |
2m |
3f |
||||||
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
Males n=10 |
Females n=12 |
||
Mesenteric lymph nodes |
congestion |
- |
- |
- |
- |
- |
- |
1 [6] |
一 |
hyperplasia |
1 [6] |
- |
- |
- |
- |
- |
1 [6] |
一 |
|
lymphocytic tissue depletion |
- |
1 [17] |
- |
- |
- |
- |
- |
一 |
|
[]-computer numbers of animals in which the histopathological lesions were found n - number of tested animals f - full histopathology
m - histopathology of thyroid, testicles, ovaries with oviducts, epididymides, prostate, seminal vesicles, coagulating glands and gross lesions were conducted
Table 58. VL3. Main study. Histopathological lesions: group 0 SAT, group 3 SAT.
Examined organ / tissue |
Type of change |
GROUP / sex / number of animals / computer numbers of animals |
|||
0 SATf |
3 SATf |
||||
Males n=10 |
Females n=10 |
Males n=10 |
Females n=10 |
||
Pituitary |
hypertrophy |
- |
1 [44] |
- |
- |
microcyst |
2 [35,40] |
- |
- |
- |
|
Thymus |
congestion |
2 [37,38] |
- |
2 [36,38] |
1 [49] |
effusion |
1 [31] |
3 [45,48,49] |
2 [34,36] |
4 [42,43,48,49] |
|
Heart |
congestion |
1 [36] |
- |
3 [36,37,38] |
- |
erythrocytorrhagia |
1 [36] |
- |
4 [36,37,38,40] |
1 [46] |
|
foci of fibrosis |
2 [36,38] |
- |
3 [34,37,40] |
- |
|
left ventricular myocardium hypertrophy |
3 [36,37,40] |
- |
2 [34,37] |
- |
|
Table 58 cont. VL3. Main study. Histopathological lesions: group 0 SAT, group 3 SAT.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
Examined |
Type of change |
0 SATf |
3 SATf |
||
organ / tissue |
Males n=10 |
Females n=10 |
Males n=10 |
Females n=10 |
|
|
congestion |
4 [32,35,39,40] |
- |
4 [34,36,37,40] |
1 [47] |
|
erythrocytorrhagia |
8 [31,32,33,34,35, 37,39,40] |
9 [41,43,44,45,46, 47,48,49,50] |
9 [31,32,33,34,36, 37,38,39,40] |
9 [41,42,43,44,45, 46,47,48,49] |
|
edema |
3 [31,32,33] |
- |
4 [32,34,37,40] |
1 [47] |
Lungs |
emphysema |
5 [31,32,36,37,39] |
4 [43,44,48,50] |
4 [31,33,35,40] |
4 [46,48,49,50] |
|
foamy cell infiltration |
- |
1 [50] |
1 [36] |
1 [41] |
|
lymphocyte infiltration |
1 [40] |
- |
- |
- |
|
inflammatory infiltration |
1 [35] |
- |
- |
- |
|
foci of fibrosis |
1 [40] |
- |
- |
- |
|
congestion |
1 [35] |
- |
1 [37] |
- |
Liver |
effusion |
- |
- |
1 [37] |
- |
|
Browicz-Kupffer cell proliferation |
1 [34] |
1 [46] |
- |
- |
|
focus of fibrosis |
- |
1 [46] |
- |
- |
Table 58 cont. VL3. Main study. Histopathological lesions: group 0 SAT, group 3 SAT.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
Examined |
Type of change |
0 SATf |
3 SATf |
||
organ / tissue |
Males n=10 |
Females n=10 |
Males n=10 |
Females n=10 |
|
Spleen |
congestion |
1 [37] |
- |
1 [37] |
- |
hemosiderin deposits |
- |
- |
1 [35] |
2 [47,48] |
|
|
islets of Langerhans congestion |
2 [35,37] |
- |
2 [35,37] |
2 [45,48] |
Pancreas |
islets of Langerhans hypertrophy |
7 [33,34,35,36,37, 38,401 |
2 [41,50] |
3 [31,35,37] |
3 [42,45,48] |
lymphocyte infiltration |
2 [35,36] |
1 [41] |
- |
- |
|
|
foci of atrophy of lobules with fibrosis |
- |
- |
1 [35] |
- |
|
ducts proliferation |
- |
- |
1 [37] |
- |
Jejunum |
lymphatic follicle reactive hyperplasia |
- |
1 [43] |
- |
- |
Cecum |
lymphatic follicle reactive hyperplasia |
1 [34] |
- |
2 [34,36] |
3 [41,42,45] |
Colon |
lymphatic follicle reactive hyperplasia |
- |
1 [50] |
- |
- |
lymphocyte infiltration |
- |
1 [48] |
- |
1 [50] |
|
Table 58 cont. VL3. Main study. Histopathological lesions: group 0 SAT, group 3 SAT.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
Examined |
Type of change |
0 SATf |
3 SATf |
||
organ / tissue |
Males n=10 |
Females n=10 |
Males n=10 |
Females n=10 |
|
|
glomeruli congestion |
5 [35,36,37,38,40] |
4 [41,45,47,50] |
5 [32,33,34,35,37] |
6 [43,44,45,47,48, 50] |
Kidneys |
glomeruli hypertrophy |
6 [32,35,36,37,38, 40] |
4 [41,45,47,50] |
6 [32,33,34,35,37, 39] |
5 [44,45,47,48,50] |
glomeruli atrophy |
3 [32,37,38] |
2 [45,47] |
3 [34,35,39] |
3 [44,48,50] |
|
|
Bowman's capsule extension |
1 [32] |
- |
1 [39] |
- |
|
retention microcysts |
1 [37] |
2 [45,47] |
- |
- |
Urinary bladder |
lymphocyte infiltration |
- |
- |
1 [34] |
- |
Prostate |
congestion |
1 [38] |
- |
- |
- |
lymphocyte infiltration |
3 [32,34,38] |
- |
2 [35,40] |
- |
|
Ovaries |
congestion |
- |
1 [43] |
- |
- |
Table 58 cont. VL3. Main study. Histopathological lesions: group 0 SAT, group 3 SAT.
|
|
GROUP / sex / number of animals / computer numbers of animals |
|||
Examined |
Type of change |
0 SATf |
3 SATf |
||
organ / tissue |
Males n=10 |
Females n=10 |
Males n=10 |
Females n=10 |
|
|
congestion |
- |
- |
1 [34] |
1 [41] |
Mandibular |
erythrocytorrhagia |
- |
- |
- |
1 [41] |
lymph nodes |
effusion |
- |
- |
1 [34] |
1 [43] |
|
lymphocytic tissue reactive hyperplasia |
- |
1 [42] |
- |
- |
Nodule (gross lesion isolated from the subcutaneous tissue of cervical area above the salivary gland) |
lipoma |
- |
- |
1 [31] |
- |
Nodule (gross lesion isolated from the fat tissue of the left ovary area) |
lipoma |
- |
- |
- |
1 [41] |
Nodule (gross lesion isolated from the fat tissue of the pelvis cavity inlet area) |
lipoma |
- |
- |
- |
1 [46] |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- 250 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
- Quality of whole database:
- The study has a Klimisch score of 1.
- Organ:
- bladder
- kidney
Repeated dose toxicity: inhalation - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: inhalation - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
Based on the available information, the substance is not classified for repeated dose toxicity (STOT RE) in accordance with CLP Regulation (EU) No. 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
