Report, BSL BIOSERVICE Study No. 112936 Version: Final
1.
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Contents page
1. Contents ....................................................................................................................................... 2 2. List of Tables and Figures .......................................................................................................... 4 2.1.
List of Tables ................................................................................................................................................... 4
3. Preface .......................................................................................................................................... 6 3.1. 3.2. 3.3. 3.4.
Abbreviations .................................................................................................................................................. 6 General ........................................................................................................................................................... 8 Project Staff .................................................................................................................................................... 8 Schedule .......................................................................................................................................................... 8
4. Project Staff Signatures ............................................................................................................ 10 5. Quality Assurance ..................................................................................................................... 11 5.1. 5.2.
Guidelines ..................................................................................................................................................... 11 Archiving ....................................................................................................................................................... 11
6. Summary .................................................................................................................................... 12 6.1. 6.2.
Summary Results ........................................................................................................................................... 12 Conclusion .................................................................................................................................................... 12
7. Introduction ............................................................................................................................... 13 7.1. 7.2.
Justification for the Selection of the Test System .......................................................................................... 13 Justification for the Selection of the Test Method ......................................................................................... 13
8. Materials and Methods ............................................................................................................. 14 8.1. 8.2. 8.3. 8.4. 8.5. 8.5.1. 8.6. 8.7. 8.8. 8.9. 8.10. 8.11. 8.12. 8.13. 8.14. 8.15. 8.16.
Characterisation of the Test Item .................................................................................................................. 14 Characterisation of the Vehicle .................................................................................................................... 14 Preparation of the Test Item Formulations ................................................................................................... 14 Dose Formulation Analysis ........................................................................................................................... 14 Test System .................................................................................................................................................... 14 Housing and Feeding Conditions ............................................................................................................ 15 Number and Sex of Animals .......................................................................................................................... 15 Preparation of the Animal............................................................................................................................. 15 Administration of Doses ................................................................................................................................ 15 Dosage .......................................................................................................................................................... 16 Body Weight and Food Consumption ........................................................................................................... 16 Clinical Observation ..................................................................................................................................... 16 Haematology ................................................................................................................................................. 16 Clinical Biochemistry.................................................................................................................................... 17 Pathology ...................................................................................................................................................... 17 Organ Weight ................................................................................................................................................ 18 Histopathology .............................................................................................................................................. 18
9. Results ........................................................................................................................................ 19 9.1. 9.2. 9.3. 9.4. 9.5. 9.6. 9.7. 9.8.
Mortality ....................................................................................................................................................... 19 Clinical Observation ..................................................................................................................................... 19 Body Weight and Body Weight Change ........................................................................................................ 19 Food Consumption ........................................................................................................................................ 19 Haematology ................................................................................................................................................. 19 Clinical Biochemistry.................................................................................................................................... 19 Gross Pathology............................................................................................................................................ 20 Organ Weight ................................................................................................................................................ 20
Report, BSL BIOSERVICE Study No. 112936 Version: Final
9.9.
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Histopathology .............................................................................................................................................. 20
10. Conclusion ................................................................................................................................. 21 11. Distribution of the Report ........................................................................................................ 22 12. References .................................................................................................................................. 23 12.1. 12.2.
Internal BSL BIOSERVICE SOPs ................................................................................................................. 23 Literature and Guidelines ............................................................................................................................. 23
13. Appendix .................................................................................................................................... 24 13.1. 13.2.
Appendix 1: Summary, Mean and Individual Tables .................................................................................... 24 Appendix 2: Histopathology Report .............................................................................................................. 74
Report, BSL BIOSERVICE Study No. 112936 Version: Final
2.
List of Tables and Figures
2.1.
List of Tables
page 4 of 110
page Table 1: Group and Animal Identification
16
Table 2: Haematology
17
Table 3: Clinical Biochemistry
17
Table 4: Organs to be weighed at Necropsy
18
Table 5: Mortality Table - Summary
24
Table 6: Clinical Observations - Males - Summary
25
Table 7: Clinical Observations - Females - Summary
25
Table 8: Mean Body Weight (g) - Males
26
Table 9: Mean Body Weight (g) - Females
26
Table 10: Mean Body Weight Gain (g/week) - Males
27
Table 11: Mean Body Weight Gain (g/week) - Females
27
Table 12: Mean Food Consumption (g/day) - Males
28
Table 13: Mean Food Consumption (g/day) - Females
28
Table 14: Mean Haematology - Males
29
Table 15: Mean Haematology - Females
29
Table 16: Mean Clinical Biochemistry - Males
30
Table 17: Mean Clinical Biochemistry - Females
31
Table 18: Macroscopic Findings - Males - Summary
32
Table 19: Macroscopic Findings - Females - Summary
32
Table 20: Absolute Mean Organ Weights (g) - Males
33
Table 21: Absolute Mean Organ Weights (g) - Females
35
Table 22: Relative Mean Organ Weights to Brain Weight (%) - Males
37
Table 23: Relative Mean Organ Weights to Brain Weight (%) - Females
39
Table 24: Relative Mean Organ Weights to Body Weight (%) - Males
41
Table 25: Relative Mean Organ Weights to Body Weight (%) - Females
43
Table 26: Individual Clinical Findings - Males
45
Table 27: Individual Clinical Findings - Females
47
Table 28: Individual Body Weight (g) - Males
49
Table 29: Individual Body Weight (g) - Females
50
Table 30: Individual Body Weight Gain (g/week) - Males
51
Table 31: Individual Body Weight Gain (g/week) - Females
52
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Table 32: Individual Food Consumption (g/day) - Males
53
Table 33: Individual Food Consumption (g/day) - Females
54
Table 34: Individual Haematology - Males
54
Table 35: Individual Haematology - Females
55
Table 36: Individual Clinical Biochemistry - Males
56
Table 37: Individual Clinical Biochemistry - Females
57
Table 38: Individual Macroscopic Findings - Males
58
Table 39: Individual Macroscopic Findings - Females
60
Table 40: Absolute Individual Organ Weights (g) - Males
62
Table 41: Absolute Individual Organ Weights (g) - Females
64
Table 42: Relative Individual Organ Weights to Brain Weight (%) - Males
66
Table 43: Relative Individual Organ Weights to Brain Weight (%) - Females
68
Table 44: Relative Individual Organ Weights to Body Weight (%) - Males
70
Table 45: Relative Individual Organ Weights to Body Weight (%) - Females
72
Report, BSL BIOSERVICE Study No. 112936 Version: Final
3.
Preface
3.1.
Abbreviations ALAT
alanine aminotransferase
Alb
albumin
ANOVA
Analysis of Variance
AP
alkaline phosphatase
Art.
Artikel (article)
ASAT
aspartate-aminotransferase
BGBl.
Bundesgesetzblatt (Federal Law Gazette)
bw
body weight
C
control
CFR
Code of Federal Regulations
Che
cholinesterase
Chol
cholesterol
Crea
creatinine
Dipl.-Biol.
Diplom Biologe (Biology Diploma)
e.g.
exempli gratia (for example)
EC
European Commission
EDTA
ethylen diamine tertraacetic acid
Ery
erythrocytes
GLP
Good Laboratory Practice
GmbH
Gesellschaft mit beschr채nkter Haftung (company with limited liability)
H&E
hematoxylin & eosin
Hb
haemoglobin
Hct
haematocrit
HD
high dose
IVC
individually ventilated cages
K
potassium
LD
low dose
Leu
leukocytes
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Report, BSL BIOSERVICE Study No. 112936 Version: Final
Lym
lymphocytes
MD
medium dose
Na
sodium
Nr.
Nummer (number)
OECD
Organisation for Economic Cooperation and Development
PLT
platelet count
QA
Quality Assurance
QAU
Quality Assurance Unit
RBC
red blood cell count
SOP
Standard Operating Procedures
SPF
specific-pathogen free
TP
total protein
WBC
white blood cells
WI
Wistar
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Report, BSL BIOSERVICE Study No. 112936 Version: Final
3.2.
3.3.
General Sponsor:
Vires5 bvba Bredabaan 926 2990 Wuustwezel Belgium
Study Monitor:
Mr Remco Schade
Test Facility:
BSL BIOSERVICE Scientific Laboratories GmbH BehringstraĂ&#x;e 6/8 82152 Planegg Germany
BSL BIOSERVICE Study No.:
112936
Test Item:
Vires5
Title:
14-Day Dose Range Finding Oral Toxicity Study in Wistar Rats with Vires5
Project Staff Study Director:
Dr. Philip Allingham
Management:
Dr. Wolfram Riedel Dr. Angela Lutterbach
Head of Quality Assurance Unit:
3.4.
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Dipl.-Biol. Uwe Hamann
Test Site 1: (Histopathology)
KALEIDIS- Consultancy in Histopathology 6 rue du Gers 68300 Saint-Louis France
Principal Investigator:
Dr. Gabriele Pohlmeyer-Esch
Test Site 2: (Tissue Processing)
Propath UK Ltd Willow Court, Netherwood Road Hereford HR2 6JU Great Britain
Principal Investigator:
Mrs Elizabeth Richards
Schedule Arrival of the Test Item: Date of Draft Study Plan: Date of Final Study Plan: Start of Experiment: End of Experiment: Start of Delegated Phase (Histopathology): End of Delegated Phase (Histopathology):
20 June 2011 28 June 2011 29 June 2011 06 July 2011 21 July 2011 25 August 2011 25 August 2011
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Date of Draft Phase Report (Histopathology): Date of Final Phase Report (Histopathology): Date of Draft Report (BSL): Date of Final Report (BSL):
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30 August 2011 22 September 2011 29 August 2011 26 September 2011
Report, BSL BIOSERVICE Study No. 112936 Version: Final
5.
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Quality Assurance This study was performed in conformity with internal quality assurance regulations, on the basis of GLP regulations, but was not audited by the quality assurance unit. Therefore, it does not have a GLP status. The test facility BSL BIOSERVICE Scientific Laboratories GmbH is certified according to the Principles of Good Laboratory Practice and accredited according to 90/385/EWG [1], 93/42/EWG [2] and DIN EN ISO/IEC 17025:2000 [3].
5.1.
Guidelines This study followed the procedures indicated by internal BSL BIOSERVICE SOPs and the following internationally accepted guidelines and recommendations: First Addendum to OECD Guidelines for Testing of Chemicals, Section 4, No. 407, “Repeated Dose 28-Day Oral Toxicity Study in Rodents”, adopted 03 October, 2008 [4]. Commission Directive 2001/59/EC 06 August 2001 [5] Commission Regulation (EC) No. 440/2008, L 142, Annex Part B, May 30, 2008 [6] “OECD Series on principles of Good Laboratory Practice and compliance monitoring” Document No 13 ENV/JM/MONO (2002) 9 [7]
5.2.
Archiving All original data generated during the conduct of the study (raw data, copy of report) will be stored in the scientific archives of BSL BIOSERVICE Scientific Laboratories GmbH for 12 years after issue of the report. The remaining test item will be discarded three months after the release of the report.
Report, BSL BIOSERVICE Study No. 112936 Version: Final
6.
Summary
6.1.
Summary Results
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The aim of this study was to assess possible health hazards of the test item Vires5 which could arise from repeated exposure via oral administration to rats over a period of 14 days and also to provide a basis for the selection of dose levels for a 28- day repeated dose oral toxicity study. The test item was administered orally in graduated doses to 3 groups of male and female Wistar rats, Crl: WI(Han) (Full Barrier) by oral gavage using a gavaging canula. Each group comprised 3 males and 3 females. One group received the vehicle water which served as control. The application volume was 5 mL/kg body weight. The animals were treated with the test item 7 days per week for a period of 14 days. For each animal the individual dosing volume was calculated on the basis of the body weight most recently measured. All surviving animals were sacrificed on day 15 and were subjected to a detailed gross necropsy. All animals found dead, moribund or intercurrently sacrificed were subjected to a gross necropsy. The wet weight of the organs was taken from all terminally sacrificed animals as soon as possible. Haematological and clinical biochemistry examinations were made on blood samples obtained from animals. A histopathological evaluation was carried out on all animals of all groups which were sacrificed at the end of the treatment period. There was no test item-related mortality and no effect of Vires5 on the health condition of the animals was noted. No findings were recorded for body weight development, food consumption, haematology, and clinical biochemistry parameters analyzed. Macroscopic findings were incidental and not related to the test item. Slight differences in spleen and thymus weight are not assumed to be test item related. No histopathological findings were noted that could be related to Vires5. 6.2.
Conclusion On the basis of the present study, a 14-day Repeated dose oral toxicity study with Vires5 in male and female rats, with dose levels of 30, 300 and 1000 mg/kg/day the following conclusions can be made. None of the dose levels used in this study was associated with any sign of toxicity. Therefore the limit dose of 1000 mg/kg bw can be recommended to be used in the high dose group of higher tier studies (e.g. 28-day oral repeated dose toxicity study).
Report, BSL BIOSERVICE Study No. 112936 Version: Final
7.
Introduction
7.1.
Justification for the Selection of the Test System
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This test is performed on the rat. Although several mammalian species may be used, the rat is the preferred rodent species. This study provides information on the possible health hazards which could arise from repeated exposure via oral administration to rats over a period of 14 days. As a dose range finding study, it should provide a basis for the selection of dose levels to be used in a 28-day repeated dose oral (gavage) toxicity study. The test item is administered daily in graduated doses to 3 groups of test animals, one dose level per group for a treatment period of 14 days. During the treatment period, the animals are observed each day for signs of toxicity. Animals which die or are humanely sacrificed in a condition of impending or predictable death are subjected to necropsy. At the conclusion of the test, the surviving animals are sacrificed and subjected to necropsy. 7.2.
Justification for the Selection of the Test Method No validated in vitro method is available for assessing systemic toxicity after repeated exposure.
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8.
Materials and Methods
8.1.
Characterisation of the Test Item The test item and the information concerning the test item were provided by the sponsor. All data related to the test item are the responsibility of the sponsor and have not been verified by the test facility.
8.2.
Name:
Vires5
Product:
Enhanced Redox Water
Batch No.:
23105
Physical State at RT:
Liquid
Density:
1.002 kg/l (4°C)
Colour:
Colourless
Date of production:
31 March 2011
Expiry Date:
No expiry date is available
Storage Conditions:
2-8°C
Safety Precautions:
Eye protection was recommended
Characterisation of the Vehicle The test item was suspended in aqua ad injectionem (sterile water). The test item formulation was prepared freshly on each administration day before the administration procedure.
8.3.
Preparation of the Test Item Formulations The test item was weighed into a tared plastic vial on a suitable precision balance and the vehicle (aqua ad injectionem) was added to give the appropriate final concentration of the test item. Afterwards, the test sample was thoroughly mixed. The vehicle was selected as suggested by the sponsor and on the basis of the test item’s characteristics.
8.4.
Dose Formulation Analysis A dose formulation analysis was not performed in this study.
8.5.
Test System Species/strain:
healthy Wistar rats, Crl: WI(Han) (Full Barrier)
Source:
Charles River, 97633 Sulzfeld, Germany
Sex:
male and female; the female animals were non-pregnant and nulliparous.
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Age at the first administration:
8-9 weeks old
Body weight at the beginning of the study:
males: 232 - 250 g; females: 158 - 181 g
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the German Act on Animal Welfare [8] the animals are bred for experimental purposes. 8.5.1. Housing and Feeding Conditions
8.6.
-
Full barrier in an air-conditioned room
-
Temperature: 22 ± 3 °C
-
Relative humidity: 55 ± 10%
-
Artificial light, sequence being 12 hours light, 12 hours dark
-
Air change: 10 x / hour
-
Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1018)
-
Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
-
The animals were housed individually in IVC cages, type III H, polysulphone cages on Altromin saw fiber bedding (lot no. 040311)
-
Certificates of food, water and bedding are filed at BSL BIOSERVICE.
-
Adequate acclimatisation period (at least five days)
Number and Sex of Animals 12 males and 12 female animals were used for the study. It included 1 control (C) and 3 dose groups (LD, MD, HD), each group containing 3 males and 3 females.
8.7.
Preparation of the Animal Prior to the start of the treatment period a detailed clinical observation outside the home cage was made. Animals showing pathological signs before the first administration were excluded from the study. Supplementary animals from the same delivery were provided in exchange. Before the first administration all animals used for the study were weighed and assigned to the experimental groups with achieving a most homogenous variation in body weight throughout the groups of males and females.
8.8.
Administration of Doses The animals were treated with the test item or vehicle on 7 days per week for a period of 14 days. The test item formulation or vehicle was administered at a single dose to the animals by oral gavage. For each animal the individual dosing volume was calculated on the basis of the body weight most recently measured.
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8.9.
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Dosage In consultation with the sponsor the following doses (Table 1) were selected for the 3 dose groups (LD, MD, HD) and 1 control group (C): Table 1: Group and Animal Identification
Group No.
Group Name
Animal No.
Dose [mg/kg bw] M
F
1
C
0
1-3
13-15
2
LD
30
4-6
16-18
3
MD
300
7-9
19-21
4
HD
1000
10-12
22-24
C = control, LD = low dose, MD = medium dose, HD = high dose
8.10. Body Weight and Food Consumption The body weight was recorded once before assignment to the experimental groups and on study days 1, 4, 8, 11 and 14 during the treatment period as well as on the day of necropsy. Food consumption was measured on study days 1, 8 and 14 for each animal. 8.11. Clinical Observation Animals were observed for clinical signs during the entire treatment period of 14 days. General clinical observations were made at least once a day, approximately at the same time each day and considering the peak period of anticipated effects after dosing. The health condition of the animals was recorded. At least once daily all animals were observed for morbidity and mortality. 8.12. Haematology To investigate major toxic effects on blood cells, the following haematological examinations (Table 2) were made. After overnight fasting of the animals, blood from the abdominal aorta was collected in EDTA-coated tubes prior to or as part of the sacrifice of the animals.
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Table 2: Haematology Parameter
Instrument
Units
haematocrit value (HCT)
ADVIA®120 (Siemens)
%
haemoglobin content (Hb)
ADVIA®120 (Siemens)
g/dL
red blood cell count (RBC)
ADVIA®120 (Siemens)
1012/L
platelet count (PLT)
ADVIA®120 (Siemens)
109/L
white blood cells (WBC)
ADVIA®120 (Siemens)
109/L
8.13. Clinical Biochemistry To investigate major toxic effects, the following clinical biochemistry examinations (Table 3) were made. After overnight fasting of the animals, blood from the abdominal aorta was collected in serum separator tubes prior to or as part of the sacrifice of the animals. Table 3: Clinical Biochemistry Parameter
Instrument
Units
alanine aminotransferase (ALAT)
Synchron Cx 5 Beckman
U/L
aspartate-aminotransferase (ASAT)
Synchron Cx 5 Beckman
U/L
alkaline phosphatase (AP)
Synchron Cx 5 Beckman
U/L
creatinine (Crea)
Synchron Cx 5 Beckman
µmol/L
total protein (TP)
Synchron Cx 5 Beckman
g/L
albumin (Alb)
Synchron Cx 5 Beckman
g/L
urea
Synchron Cx 5 Beckman
mmol/L
total cholesterol (Chol)
Synchron Cx 5 Beckman
mmol/L
sodium (Na)
Synchron Cx 5 Beckman
mmol/L
potassium (K)
Synchron Cx 5 Beckman
mmol/L
8.14. Pathology On study day 15, all surviving animals of the study were subjected to a detailed gross necropsy which included careful examination of the external surface of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents. All macroscopic findings were recorded and organs showing gross abnormalities were preserved in neutral buffered formalin.
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All animals found moribund and/or intercurrently sacrificed were subjected to a gross necropsy. 8.15. Organ Weight The wet weight of the following organs (Table 4) was taken from all sacrificed animals as soon as possible. Paired organs were weighed separately. Table 4: Organs to be weighed at Necropsy Tissue/Organ
Tissue/Organ
liver
ovaries
kidneys
uterus with cervix (females)
adrenals
thymus
testes
spleen
epididymides
heart
Prostate and seminal vesicles with coagulating gland (as a whole)
brain
8.16. Histopathology A histopathological evaluation of stomach as well as small (duodenum, jejunum, ileum) and large (caecum, colon) intestine (after the preparation of paraffin sections and haematoxylin-eosin staining) was carried out on all animals of all groups. The histological processing of tissues to microscope slides was performed at the GLPcertified contract laboratory Propath UK Ltd, Willow Court, Netherwood Road, Hereford HR2 6JU, Great Britain (test site for tissue processing). The histopathological evaluation were performed at the GLP-certified contract laboratory KALEIDIS – Consultancy in Histopathology (test site for histopathology), 6 rue du Gers, 68300 Saint-Louis, France. Blocking, embedding, cutting, H&E staining and scientific slide evaluation were performed according to the corresponding SOP´s of the test sites. The principal histopathological investigator provided the histopathology results to the study director by e-mail and sent a pathology phase report to the study director upon the completion of the study.
Report, BSL BIOSERVICE Study No. 112936 Version: Final
9.
Results
9.1.
Mortality
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For a summary see Table 5. There was no test item related mortality in this study. 9.2.
Clinical Observation For a summary see Table 6; for a detailed description of the individual findings see Tables 26 and 27. On the day of necropsy piloerection was noted in 2 female animals of the high dose group. This was, however, not associated with any other pathological signs. None of the male animals of this group was affected. Thus this slight finding is not assumed to be related to the test item. With the exception of transient nasal discharge in one animal of the mid dose group, which is not assumed to be test item related, there were no clinical findings in this study
9.3.
Body Weight and Body Weight Change For group means see Tables 8, 9, 10 and 11; for individual data see Tables 28, 29, 30 and 31. Vires5 did not have any effect on body weight development in this study. Throughout the treatment period body weights of male and female animals of all groups were in the normal range of variation for this strain and age. There were no considerable differences in body weight between any of the dose groups and the control group.
9.4.
Food Consumption For group means see Tables 12 and 13; for individual data see Tables 32 and 33. Vires5 did not affect food consumption in this study. There were no biologically relevant differences in food intake between any of the dose groups and the control group.
9.5.
Haematology For group means see Tables 14 and 15; for individual data see Tables 34 and 35. All haematological parameters analyzed in this study were in the normal range of biological variation for this strain. There were no considerable differences, neither in red cell parameters (RBC, haematocrit, haemoglobine) nor in white blood cells or platelets when comparing dose groups to control group.
9.6.
Clinical Biochemistry For group means see Tables 16 and 17; for individual data see Tables 36 and 37.
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All clinical pathology parameters analyzed at the end of the treatment period were within the normal range of variation for this strain. There were no considerable differences between dose groups and control group. 9.7.
Gross Pathology For a summary see Tables 18 and 19; for a detailed description of the individual findings see Tables 38 and 39. After 14 days of oral treatment with Vires5 only one or few mild findings were noted that are assumed to be incidental and not related to the test item. One male animal of the low dose group had a discolored red stomach, which was not associated with any histopathological findings in this organ. On the epididymides of two male animals (one from the low dose group and one from the high dose group), yellow foci were found – a common background finding in this strain. One male animal of the high dose group had blood on the surface of the brain and 2 male animals (one of the medium dose group and one of the high dose group) had a cyst on the adrenals. These single mild findings are not assumed to be related to the test item. One female control animal of the low dose group had a thickened mucosa in the stomach, which was not associated with any histopathological findings in this organ.
9.8.
Organ Weight For group means see Tables 20 to 25; for individual data see Tables 40 to 45. The absolute and relative spleen weight of the spleens of the male animals of the high dose group was slightly lower than in controls. As this slight difference in spleen weight was not observed in female animals and due to the low number of animals, this is not assumed to be related to the test item. A tendency towards a lower thymus weight was also observed in Vire5 treated male animals of this study, however, not dose-dependently. Due to the slight degree, the low number of animals and the fact that female animals were not affected, a relation to the test item is doubtful. Besides, there were no considerable differences in organ weights of Vires5 treated animals when compared to controls.
9.9.
Histopathology Histopathological findings noted in the organs evaluated were very few and none of them was considered to be test item-related.
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10. Conclusion On the basis of the present study, a 14-day Repeated dose oral toxicity study with Vires5 in male and female rats, with dose levels of 30, 300 and 1000 mg/kg/day the following conclusions can be made. None of the dose levels used in this study was associated with any sign of toxicity. Therefore the limit dose of 1000 mg/kg bw can be recommended to be used in the high dose group of higher tier studies (e.g. 28-day oral repeated dose toxicity study).
Report, BSL BIOSERVICE Study No. 112936 Version: Final
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11. Distribution of the Report 1 original (paper):
Sponsor
1 copy (paper):
BSL BIOSERVICE
1 copy (electronic):
Sponsor
Report, BSL BIOSERVICE Study No. 112936 Version: Final
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12. References 12.1. Internal BSL BIOSERVICE SOPs Standard Operating Procedures (SOP), No. 11-4-3
12.2. Literature and Guidelines [1]
Richtlinie 90/385/EWG des Rates vom 20. Juni 1990 zur Angleichung der Rechtsvorschriften der Mitgliedstaaten über aktive implantierbare medizinische Geräte, Amtsblatt Nr. L 189 vom 20/07/1990 S. 0017 - 0036
[2]
Richtlinie 93/42/EWG des Rates vom 14. Juni 1993 über Medizinprodukte, Amtsblatt Nr. L 169 vom 12/07/1993 S. 0001 - 0043
[3]
DIN EN ISO/IEC 17025: 2005: Allgemeine Anforderungen an die Kompetenz von Prüf- und Kalibrierlaboratorien, DIN Deutsches Institut für Normung e.V., Berlin, August 2005
[4]
First Addendum to OECD Guidelines for Testing of Chemicals, Section 4, No. 407, “Repeated Dose 28-Day Oral Toxicity Study in Rodents”, adopted 03 October, 2008
[5]
Commission Directive 2001/59/EC adapting to technical progress for the 28th time Council Directive 67/548/EC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances, 06 August 2001 (Official Journal of the European Communities no. L 225/1, 21 August 2001)
[6]
Commission Regulation (EC) No 440/2008, L 142, Annex Part B of 30 May 2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH)
[7]
OECD Series on principles of Good Laboratory Practice and compliance monitoring Document No 13 ENV/JM/MONO (2002) 9
[8]
German Animal Welfare Act, Art. 9.2, No. 7 (Deutsches Tierschutzgesetz, 24. Juli 1972 (BGBl. I S. 1277), Inkrafttreten der letzten Änderung: 22. Juli 2009, (Art. 2 ÄndG vom 15. Juli 2009)
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13. Appendix 13.1. Appendix 1: Summary, Mean and Individual Tables Table 5:
Mortality Table - Summary Animals
Dose Group
Male
Female
C
0/3
0/3
LD
0/3
0/3
MD
0/3
0/3
HD
0/3
0/3
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Table 6:
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Clinical Observations - Males - Summary
Clinical Finding
C
LD
MD
HD
Total number of animals examined
3
3
3
3
Nasal discharge
‐
1
‐
‐
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Table 7:
Clinical Observations - Females - Summary
Clinical Finding
C
LD
MD
HD
Total number of animals examined
3
3
3
3
Slight piloerection
‐
‐
‐
2
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Table 8:
Mean Body Weight (g) - Males Study Day
Group
C
LD
MD
HD
page 26 of 110
1
4
8
11
14
Mean
247.33
269.33
289.37
298.23
308.97
SD
5.03
2.31
4.47
3.82
8.98
N
3
3
3
3
3
Mean
252.33
269.00
295.03
304.40
314.43
SD
4.51
7.00
7.77
6.63
9.21
N
3
3
3
3
3
%
102.02
99.88
101.96
102.07
101.77
Mean
245.33
267.00
286.57
296.27
306.23
SD
3.79
6.08
4.81
8.66
8.52
N
3
3
3
3
3
%
99.19
99.13
99.03
99.34
99.12
Mean
249.00
266.00
281.80
290.57
297.27
SD
6.24
7.21
6.43
4.95
5.22
N
3
3
3
3
3
%
100.67
98.76
97.39
97.43
96.21
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Table 9:
Mean Body Weight (g) - Females Study Day
Group
C
LD
MD
HD
1
4
8
11
14
Mean
169.57
174.33
181.00
178.47
185.20
SD
4.53
5.69
3.00
3.82
4.53
N
3
3
3
3
3
Mean
166.63
172.33
178.33
183.47
185.40
SD
5.15
13.58
13.58
11.38
10.72
N
3
3
3
3
3
%
98.27
98.85
98.53
102.80
100.11
Mean
165.13
172.33
175.67
176.20
180.30
SD
4.25
7.64
5.51
5.78
4.71
N
3
3
3
3
3
%
97.39
98.85
97.05
98.73
97.35
Mean
171.37
178.33
182.33
182.87
186.97
SD
1.14
3.51
2.52
6.45
7.07
N
3
3
3
3
3
%
101.06
102.29
100.74
102.47
100.95
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 27 of 110
Table 10: Mean Body Weight Gain (g/week) - Males Group C
LD
MD
HD
Day 1-4
Day 4-8
Day 8-11 Day 11-14 Day 1-14
Mean
22.00
20.03
8.87
10.73
61.63
SD
3.46
3.10
4.86
5.26
8.22
N
3
3
3
3
3
Mean
16.67
26.03
9.37
10.03
62.10
SD
3.51
1.47
6.05
3.76
7.46
N
3
3
3
3
3
Mean
21.67
19.57
9.70
9.97
60.90
SD
2.31
1.60
4.46
2.03
5.38
N
3
3
3
3
3
Mean
17.00
15.80
8.77
6.70
48.27
SD
7.00
1.04
2.25
1.30
5.75
N
3
3
3
3
3
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Table 11: Mean Body Weight Gain (g/week) - Females Group C
LD
MD
HD
Day 1-4
Day 4-8
Day 8-11 Day 11-14 Day 1-14
Mean
4.77
6.67
-2.53
6.73
15.63
SD
2.77
5.69
5.19
6.63
4.02
N
3
3
3
3
3
Mean
5.70
6.00
5.13
1.93
18.77
SD
8.61
0.00
2.80
0.76
5.58
N
3
3
3
3
3
Mean
7.20
3.33
0.53
4.10
15.17
SD
3.64
2.31
0.93
4.09
3.66
N
3
3
3
3
3
Mean
6.97
4.00
0.53
4.10
15.60
SD
3.89
1.00
3.99
2.29
6.77
N
3
3
3
3
3
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 28 of 110
Table 12: Mean Food Consumption (g/day) - Males Group C
LD
MD
HD
Day 1-7
Day 7-14
Day 1-14
Mean
22.90
26.22
24.56
SD
1.12
1.63
1.37
N
3
3
3
Mean
23.68
27.15
25.41
SD
0.38
0.88
0.45
N
3
3
3
%
103.39
103.54
103.47
Mean
24.35
26.41
25.38
SD
1.43
2.39
1.91
N
3
3
3
%
106.32
100.74
103.34
Mean
22.55
25.23
23.89
SD
1.88
1.32
1.60
N
3
3
3
%
98.48
96.22
97.28
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Table 13: Mean Food Consumption (g/day) - Females Group C
LD
MD
HD
Day 1-8
Day 8-14
Day 1-14
Mean
13.57
15.20
14.38
SD
0.31
1.09
0.61
N
3
3
3
Mean
14.00
15.39
14.69
SD
1.06
1.30
1.18
N
3
3
3
%
103.12
101.25
102.14
Mean
13.48
15.38
14.43
SD
1.01
2.05
1.53
N
3
3
3
%
99.33
101.19
100.31
Mean
13.94
15.90
14.92
SD
0.30
0.62
0.45
N
3
3
3
%
102.74
104.61
103.72
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 29 of 110
Table 14: Mean Haematology - Males RBC
Group
C
LD
MD
HD
HB
HCT
PLT
WBC
9
10 /L
g/dL
%
10 /L
10 9/L
Mean
8.57
16.05
47.25
1095.50
6.45
SD
0.07
0.35
0.49
3.54
2.50
N
2
2
2
2
2
Mean
8.20
15.43
45.53
1098.00
7.02
SD
0.23
0.47
2.11
48.07
2.59
N
3
3
3
3
3
Mean
8.38
15.93
46.70
948.00
4.78
SD
0.33
0.32
1.25
139.34
0.14
Units
12
N
3
3
3
3
3
Mean
9.30
16.75
49.80
1087.00
6.95
SD
0.02
0.35
1.41
39.60
0.15
N
2
2
2
2
2
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Table 15: Mean Haematology - Females RBC
Group
C
LD
MD
HD
HB
HCT
PLT
WBC
9
10 /L
g/dL
%
10 /L
10 9/L
Mean
8.02
14.67
43.57
915.33
3.15
SD
0.69
0.42
1.88
92.65
0.09
N
3
3
3
3
3
Mean
8.70
15.70
46.85
1042.50
2.54
SD
0.23
0.42
0.07
98.29
0.07
N
2
2
2
2
2
Mean
8.30
15.07
44.33
888.33
3.47
SD
0.16
0.47
0.55
48.95
0.35
N
3
3
3
3
3
Mean
8.35
15.07
45.33
903.00
3.14
SD
0.62
0.60
1.60
59.10
0.15
N
3
3
3
3
3
Units
12
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 30 of 110
Table 16: Mean Clinical Biochemistry - Males Group
C
LD
MD
HD
ASAT
ALAT
AP
Crea
TP
Alb
Urea
Chol
Na
K
U/L
U/L
U/L
Âľmol/L
g/L
g/L
mmol/L
mmol/L
mmol/L
mmol/L
Mean
35.30
20.93
247.5
31.96
60.10
34.43
8.18
0.96
150.0
4.04
SD
3.67
5.56
65.8
3.01
3.15
2.15
0.51
0.11
2.0
0.64
N
3
3
2
3
3
3
3
3
3
3
Mean
29.33
19.70
187.7
27.87
58.50
32.03
7.96
0.77
151.0
4.39
SD
4.30
1.31
40.5
1.71
1.21
0.84
0.37
0.03
0.0
0.41
N
3
3
3
3
3
3
3
3
3
3
Mean
30.40
20.53
188.0
28.16
58.13
33.43
7.56
0.85
151.0
3.77
SD
3.67
2.15
45.9
2.24
0.57
0.15
0.61
0.25
3.6
0.21
Units
N
3
3
3
3
3
3
3
3
3
3
Mean
36.10
23.35
182.5
29.99
59.70
33.80
8.23
0.81
153.5
4.47
SD
1.27
1.20
20.5
1.41
0.28
0.00
0.09
0.17
0.7
0.30
N
2
2
2
2
2
2
2
2
2
2
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 31 of 110
Table 17: Mean Clinical Biochemistry - Females Group
C
LD
MD
HD
ASAT
ALAT
AP
Crea
TP
Alb
Urea
Chol
Na
K
U/L
U/L
U/L
Âľmol/L
g/L
g/L
mmol/L
mmol/L
mmol/L
mmol/L
Mean
44.23
13.93
67.33
33.52
51.67
32.67
7.18
0.57
144.33
3.74
SD
3.50
1.59
21.13
1.68
1.40
1.15
0.17
0.05
0.58
0.56
N
3
3
3
3
3
3
3
3
3
3
Mean
37.97
14.50
68.00
31.77
53.53
33.70
6.19
0.53
145.67
3.91
SD
2.38
1.00
3.00
9.71
2.55
1.30
0.55
0.08
1.53
0.60
N
3
3
3
3
3
3
3
3
3
3
Mean
31.07
14.17
76.00
25.32
54.47
33.93
7.31
0.56
144.67
3.40
SD
1.10
1.98
21.00
3.86
2.40
0.76
1.24
0.06
2.52
0.35
Units
N
3
3
3
3
3
3
3
3
3
3
Mean
37.87
14.57
73.67
34.02
55.07
33.43
8.10
0.59
145.00
3.49
SD
3.60
0.51
20.26
5.27
0.61
1.63
0.37
0.08
2.65
0.25
N
3
3
3
3
3
3
3
3
3
3
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 32 of 110
Table 18: Macroscopic Findings - Males - Summary Day of sacrifice: 15 Finding
C
LD
MD
HD
Total number of animals examined
3
3
3
3
Stomach
discolored red
‐
1
‐
‐
Epididymides
yellow foci
‐
1
‐
1
Brain
Blood at surface
‐
‐
‐
1
Adrenals
Cyst
‐
‐
1
1
Organ
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Table 19: Macroscopic Findings - Females - Summary Day of sacrifice: 15 Finding
C
LD
MD
HD
Total number of animals examined
3
3
3
3
Stomach
Mucosa thickened
1
‐
‐
‐
Lung
discolored dark
‐
‐
‐
‐
Duodenum
discolored red
‐
‐
‐
‐
Organ
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 33 of 110
Table 20: Absolute Mean Organ Weights (g) - Males
Body weight
Brain
Mean
287
2.046
8.653
SD
4.9
0.103
N
3
Mean SD
Group
C
LD
MD
HD
Liver
Kidneys
Heart
Adrenals
Spleen
left
right
total
left
right
total
0.909
1.135
1.143
2.278
0.0376
0.0364
0.0740
0.738
0.074
0.047
0.014
0.017
0.031
0.0036
0.0040
0.0074
0.082
3
3
3
3
3
3
3
3
3
3
293
1.933
8.754
1.042
1.097
1.156
2.253
0.0383
0.0369
0.0752
0.703
7.1
0.111
0.208
0.063
0.062
0.076
0.136
0.0028
0.0045
0.0065
0.030
N
3
3
3
3
3
3
3
3
3
3
3
%
101.9
94.5
101.2
114.7
96.6
101.1
98.9
102.0
101.3
101.6
95.1
Mean
285
2.024
8.523
1.028
1.183
1.247
2.431
0.0374
0.0319
0.0693
0.743
SD
10.1
0.071
0.728
0.080
0.110
0.116
0.218
0.0043
0.0049
0.0082
0.068
N
3
3
3
3
3
3
3
3
3
3
3
%
99.1
99.0
98.5
113.2
104.2
109.1
106.7
99.5
87.6
93.7
100.6
Mean
280
2.021
10.226
0.927
1.184
1.163
2.348
0.0367
0.0366
0.0733
0.620
SD
4.0
0.107
2.682
0.036
0.084
0.031
0.111
0.0047
0.0049
0.0094
0.027
N
3
3
3
3
3
3
3
3
3
3
3
%
97.6
98.8
118.2
102.0
104.3
101.8
103.0
97.6
100.5
99.1
83.9
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 34 of 110
Table 20: Absolute Mean Organ Weights (g) - Males (Continued) Group
C
LD
MD
HD
Testes
Thymus
Epididymides
Prostate #
left
right
total
left
right
total
Mean
0.636
1.719
1.708
3.427
0.450
0.477
0.927
1.747
SD
0.109
0.058
0.065
0.108
0.015
0.017
0.032
0.119
N
3
3
3
3
3
3
3
3
Mean
0.545
1.716
1.675
3.390
0.439
0.439
0.878
1.539
SD
0.090
0.197
0.096
0.247
0.036
0.012
0.044
0.039
N
3
3
3
3
3
3
3
3
%
85.7
99.8
98.0
98.9
97.7
92.0
94.8
88.1
Mean
0.560
1.709
1.720
3.429
0.449
0.451
0.901
1.532
SD
0.057
0.130
0.123
0.252
0.043
0.039
0.081
0.051
N
3
3
3
3
3
3
3
3
%
88.0
99.4
100.7
100.1
99.9
94.6
97.2
87.7
Mean
0.541
1.636
1.674
3.310
0.469
0.484
0.953
1.866
SD
0.047
0.057
0.019
0.063
0.049
0.043
0.067
0.145
N
3
3
3
3
3
3
3
3
%
85.0
95.2
98.0
96.6
104.3
101.5
102.8
106.8
# = including coagulating glands
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 35 of 110
Table 21: Absolute Mean Organ Weights (g) - Females Body weight
Brain
Mean
173
1.781
4.920
SD
1.5
0.026
N
3
3
Mean
171
SD
6.2
Group
C
LD
MD
HD
Liver
Adrenals
Kidney
Heart left
right
total
left
right
total
0.612
0.726
0.732
1.457
0.0330
0.0373
0.0703
0.212
0.022
0.043
0.052
0.095
0.0030
0.0064
0.0085
3
3
3
3
3
3
3
3
1.815
5.410
0.573
0.701
0.752
1.453
0.0393
0.0361
0.0754
0.026
1.237
0.064
0.016
0.024
0.010
0.0040
0.0022
0.0056
N
3
3
3
3
3
3
3
3
3
3
%
99.0
101.9
110.0
93.6
96.6
102.8
99.7
119.1
96.9
107.3
Mean
170
1.809
4.886
0.613
0.698
0.763
1.461
0.0405
0.0376
0.0781
SD
5.1
0.022
0.627
0.020
0.137
0.113
0.250
0.0076
0.0055
0.0130
N
3
3
3
3
3
3
3
3
3
3
%
98.6
101.6
99.3
100.2
96.3
104.2
100.3
122.6
100.9
111.1
Mean
174
1.771
5.259
0.637
0.761
0.773
1.533
0.0442
0.0387
0.0829
SD
5.3
0.097
0.215
0.068
0.047
0.021
0.061
0.0055
0.0029
0.0052
N
3
3
3
3
3
3
3
3
3
3
%
100.8
99.4
106.9
104.1
104.9
105.6
105.2
134.0
103.8
118.0
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 36 of 110
Table 21: Absolute Mean Organ Weights (g) - Females (Continued) Group
C
LD
MD
HD
Spleen
Ovaries
Thymus
Uterus #
left
right
total
Mean
0.466
0.327
0.0524
0.0487
0.1012
0.509
SD
0.033
0.023
0.0074
0.0101
0.0175
0.105
N
3
3
3
3
3
3
Mean
0.436
0.332
0.0452
0.0418
0.0870
0.668
SD
0.058
0.056
0.0085
0.0023
0.0073
0.269
N
3
3
3
3
3
3
%
93.4
101.6
86.3
85.7
86.0
131.3
Mean
0.466
0.326
0.0459
0.0500
0.0959
0.602
SD
0.046
0.066
0.0107
0.0047
0.0135
0.369
N
3
3
3
3
3
3
%
99.9
99.7
87.5
102.5
94.8
118.2
Mean
0.474
0.339
0.0424
0.0511
0.0935
0.551
SD
0.048
0.051
0.0060
0.0022
0.0079
0.156
N
3
3
3
3
3
3
%
101.7
103.6
80.9
104.8
92.4
108.2
# = including cervix and oviduct
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 37 of 110
Table 22: Relative Mean Organ Weights to Brain Weight (%) - Males Kidneys Group
C
LD
MD
HD
Liver
Heart
Mean
423.81
SD
Adrenals Spleen
left
right
total
left
right
total
44.45
55.60
55.99
111.60
1.837
1.778
3.614
36.29
25.38
2.18
3.34
3.43
6.77
0.128
0.113
0.218
5.93
N
3
3
3
3
3
3
3
3
3
Mean
453.57
53.92
56.91
59.94
116.85
1.985
1.917
3.902
36.45
SD
16.77
0.68
5.24
5.04
10.20
0.121
0.301
0.413
2.95
N
3
3
3
3
3
3
3
3
3
%
107.0
121.3
102.3
107.0
104.7
108.1
107.9
108.0
100.4
Mean
421.61
50.93
58.55
61.64
120.19
1.851
1.576
3.427
36.71
SD
41.82
5.85
6.35
5.63
11.64
0.246
0.221
0.408
3.58
N
3
3
3
3
3
3
3
3
3
%
99.5
114.6
105.3
110.1
107.7
100.8
88.7
94.8
101.2
Mean
511.40
45.95
58.58
57.63
116.21
1.828
1.823
3.651
30.77
SD
162.20
2.48
1.78
1.58
1.67
0.336
0.332
0.660
3.02
N
3
3
3
3
3
3
3
3
3
%
120.7
103.4
105.4
102.9
104.1
99.5
102.6
101.0
84.8
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 38 of 110
Table 22: Relative Mean Organ Weights to Brain Weight (%) - Males (Continued) Group
C
LD
MD
HD
Thymus
Testes
Epididymides
Prostate #
left
right
total
left
right
total
Mean
31.28
84.25
83.70
167.95
22.01
23.33
45.34
85.56
SD
6.50
7.21
6.33
13.25
0.46
0.63
1.08
7.64
N
3
3
3
3
3
3
3
3
Mean
28.14
88.99
86.66
175.65
22.84
22.79
45.63
79.76
SD
3.39
11.32
1.67
12.94
3.05
1.97
4.90
2.73
N
3
3
3
3
3
3
3
3
%
90
106
104
105
104
98
101
93
Mean
27.63
84.44
85.00
169.44
22.24
22.36
44.60
75.76
SD
2.26
6.38
5.60
11.95
2.65
2.67
5.24
3.92
N
3
3
3
3
3
3
3
3
%
88
100
102
101
101
96
98
89
Mean
26.73
81.15
83.01
164.16
23.18
24.02
47.20
92.33
SD
0.98
5.84
5.43
10.93
1.40
2.73
3.01
4.74
N
3
3
3
3
3
3
3
3
%
85
96
99
98
105
103
104
108
# = including coagulating glands
Asterisks indicate statistically significant differences to control group C, with* p<0.05, ** p<0.01 and *** p<0.001 C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose; * = including coagulating glands
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 39 of 110
Table 23: Relative Mean Organ Weights to Brain Weight (%) - Females Group
C
LD
MD
HD
Liver
Heart
Mean
276.10
SD
8.10
N
Kidney
Adrenals
left
right
total
left
right
total
34.37
40.73
41.09
81.82
1.854
2.096
3.949
1.65
2.52
3.10
5.62
0.186
0.391
0.531
3
3
3
3
3
3
3
3
Mean
298.81
31.57
38.63
41.44
80.07
2.167
1.991
4.158
SD
72.86
3.27
0.87
1.42
0.98
0.243
0.148
0.366
N
3
3
3
3
3
3
3
3
%
108.2
91.8
94.8
100.9
97.9
116.9
95.0
105.3
Mean
270.28
33.90
38.62
42.19
80.81
2.240
2.080
4.320
SD
37.46
0.83
7.77
6.53
14.26
0.445
0.327
0.768
N
3
3
3
3
3
3
3
3
%
97.9
98.6
94.8
102.7
98.8
120.8
99.3
109.4
Mean
297.45
36.14
43.11
43.70
86.81
2.496
2.195
4.691
SD
17.34
5.34
4.55
2.34
6.89
0.236
0.282
0.364
N
3
3
3
3
3
3
3
3
%
107.7
105.1
105.8
106.4
106.1
134.6
104.8
118.8
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 40 of 110
Table 23: Relative Mean Organ Weights to Brain Weight (%) - Females (Continued) Spleen
Thymus
Mean
26.18
SD
1.72
N
Group
C
LD
MD
HD
Ovaries
Uterus #
left
right
total
18.37
2.946
2.738
5.685
28.53
1.56
0.442
0.586
1.021
5.45
3
3
3
3
3
3
Mean
23.97
18.29
2.497
2.301
4.798
36.85
SD
2.84
2.85
0.501
0.098
0.458
14.84
N
3
3
3
3
3
3
%
91.6
99.6
84.8
84.0
84.4
129.2
Mean
25.77
18.04
2.540
2.764
5.304
33.33
SD
2.77
3.80
0.615
0.289
0.801
20.61
N
3
3
3
3
3
3
%
98.4
98.2
86.2
100.9
93.3
116.8
Mean
26.94
19.26
2.400
2.892
5.292
31.28
SD
4.14
3.82
0.356
0.252
0.559
9.69
N
3
3
3
3
3
3
%
102.9
104.9
81.5
105.6
93.1
109.6
# = including cervix and oviduct
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 41 of 110
Table 24: Relative Mean Organ Weights to Body Weight (%) - Males Group
C
LD
MD
HD
Brain
Liver
Kidneys
Heart
Adrenals
Spleen
left
right
total
left
right
total
Mean
0.712
3.012
0.316
0.395
0.398
0.793
0.0131
0.0127
0.0257
0.257
SD
0.031
0.063
0.019
0.012
0.013
0.024
0.0010
0.0012
0.0022
0.032
N
3
3
3
3
3
3
3
3
3
3
Mean
0.661
2.993
0.356
0.375
0.396
0.771
0.013
0.013
0.0257
0.240
SD
0.047
0.133
0.028
0.029
0.035
0.064
0.00
0.00
0.0028
0.015
N
3
3
3
3
3
3
3
3
3
3
%
92.8
99.4
113
95
99
97
100
100
100
93
Mean
0.712
2.991
0.361
0.415
0.438
0.853
0.0131
0.0112
0.0243
0.261
SD
0.048
0.177
0.020
0.027
0.035
0.059
0.0011
0.0017
0.0024
0.031
N
3
3
3
3
3
3
3
3
3
3
%
100.0
99.3
114
105
110
108
100
89
95
102
Mean
0.721
3.641
0.331
0.423
0.415
0.838
0.0131
0.0131
0.0261
0.221
SD
0.044
0.908
0.011
0.031
0.016
0.044
0.0015
0.0016
0.0030
0.008
N
3
3
3
3
3
3
3
3
3
3
%
101.3
120.9
104.5
106.9
104.3
105.6
100.1
103.0
101.5
85.9
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 42 of 110
Table 24: Relative Mean Organ Weights to Body Weight (%) - Males (Continued) Group
C
LD
MD
HD
Testes
Thymus
Epididymides
Prostate #
left
right
total
left
right
total
Mean
0.222
0.598
0.595
1.193
0.157
0.166
0.323
0.608
SD
0.041
0.028
0.033
0.057
0.005
0.006
0.012
0.034
N
3
3
3
3
3
3
3
3
Mean
0.187
0.588
0.573
1.160
0.150
0.150
0.300
0.526
SD
0.035
0.080
0.044
0.113
0.014
0.004
0.017
0.023
N
3
3
3
3
3
3
3
3
%
84.2
98.2
96.3
97.2
96.0
90.4
93.1
86.6
Mean
0.197
0.600
0.605
1.205
0.158
0.158
0.316
0.538
SD
0.028
0.039
0.041
0.080
0.010
0.009
0.017
0.013
N
3
3
3
3
3
3
3
3
%
88.9
100.3
101.6
101.0
100.6
95.4
97.9
88.6
Mean
0.193
0.583
0.597
1.181
0.168
0.173
0.340
0.666
SD
0.019
0.013
0.008
0.005
0.0192
0.017
0.029
0.061
N
3
3
3
3
3
3
3
3
%
87.0
97.5
100.4
98.9
107.0
104.1
105.5
109.6
# = including seminal vesicles and coagulating glands
Asterisks indicate statistically significant differences to control group C, with* p<0.05, ** p<0.01 and *** p<0.001 C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose; * = including seminal vesicles and coagulating glands
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 43 of 110
Table 25: Relative Mean Organ Weights to Body Weight (%) - Females Group
C
LD
MD
HD
Brain
Liver
Adrenals
Kidney
Heart left
right
total
left
right
total
Mean
1.032
2.849
0.355
0.420
0.424
0.844
0.0191
0.0216
0.0407
SD
0.013
0.106
0.016
0.028
0.034
0.062
0.0017
0.0038
0.0050
N
3
3
3
3
3
3
3
3
3
Mean
1.062
3.170
0.335
0.410
0.440
0.850
0.0230
0.0211
0.0442
SD
0.042
0.759
0.028
0.024
0.002
0.026
0.0029
0.0014
0.0042
N
3
3
3
3
3
3
3
3
3
%
102.9
111.3
94.4
97.7
103.7
100.7
120.6
97.8
108.5
Mean
1.063
2.864
0.361
0.409
0.447
0.856
0.0237
0.0220
0.0458
SD
0.038
0.301
0.022
0.068
0.053
0.120
0.00
0.00
0.01
N
3
3
3
3
3
3
3
3
3
%
103.0
100.5
101.7
97.2
105.4
101.3
124.2
102.1
112.4
Mean
1.018
3.026
0.367
0.438
0.444
0.882
0.0255
0.0223
0.0477
SD
0.059
0.205
0.050
0.039
0.024
0.061
0.0036
0.0021
0.0043
N
3
3
3
3
3
3
3
3
3
%
98.7
106.2
103.6
104.2
104.8
104.5
133.3
103.1
117.3
Asterisks indicate statistically significant differences to control group C, with* p<0.05, ** p<0.01 and *** p<0.001 C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 44 of 110
Table 25: Relative Mean Organ Weights to Body Weight (%) - Females (Continued) Spleen
Group
C
LD
MD
HD
Ovaries
Thymus
Uterus #
left
right
total
Mean
0.270
0.189
0.0304
0.0282
0.0586
0.295
SD
0.020
0.015
0.0042
0.0057
0.0098
0.059
N
3
3
3
3
3
3
Mean
0.255
0.194
0.0264
0.0244
0.0509
0.395
SD
0.038
0.031
0.0047
0.0012
0.0035
0.169
N
3
3
3
3
3
3
%
94.4
102.5
87.0
86.6
86.8
134.0
Mean
0.274
0.192
0.0268
0.0293
0.0562
0.356
SD
0.032
0.042
0.0055
0.0029
0.0067
0.227
N
3
3
3
3
3
3
%
101.4
101.2
88.4
104.1
96.0
120.9
Mean
0.273
0.195
0.0245
0.0294
0.0539
0.319
SD
0.029
0.029
0.0041
0.0021
0.0061
0.098
N
3
3
3
3
3
3
%
101.0
102.8
80.6
104.2
92.0
108.1
# = including cervix and oviduct
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Table 26: Individual Clinical Findings - Males Dose group: C Animal No.
1 No clinical findings observed throughout the whole observation period
Animal No.
2 No clinical findings observed throughout the whole observation period
Animal No.
3 No clinical findings observed throughout the whole observation period
Dose group: LD Animal No.
4 No clinical findings observed throughout the whole observation period
Animal No.
5 No clinical findings observed throughout the whole observation period
Animal No.
6 No clinical findings observed throughout the whole observation period
page 45 of 110
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 46 of 110
Dose group: MD Animal No.
7 No clinical findings observed throughout the whole observation period
Animal No.
8 No clinical findings observed throughout the whole observation period
Animal No.
9 Clinical finding Nasal discharge
Days 10
There were no further clinical findings on any other study day for this animal
Dose group: Animal No.
HD 10 No clinical findings observed throughout the whole observation period
Animal No. 11 No clinical findings observed throughout the whole observation period
Animal No. 12 No clinical findings observed throughout the whole observation period
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Table 27: Individual Clinical Findings - Females Dose group: C Animal No.
13 No clinical findings observed throughout the whole observation period
Animal No.
14 No clinical findings observed throughout the whole observation period
Animal No.
15 No clinical findings observed throughout the whole observation period
Dose group: LD Animal No.
16 No clinical findings observed throughout the whole observation period
Animal No.
17 No clinical findings observed throughout the whole observation period
Animal No.
18 No clinical findings observed throughout the whole observation period
page 47 of 110
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 48 of 110
Dose group: MD Animal No.
19 No clinical findings observed throughout the whole observation period
Animal No.
20 No clinical findings observed throughout the whole observation period
Animal No.
21 No clinical findings observed throughout the whole observation period
Dose group: HD Animal No.
Animal No.
22 No clinical findings observed throughout the whole observation period
23 Clinical finding slight piloerection
Days 15
There were no further clinical findings on any other study day for this animal
Animal No.
24 Clinical finding Slight piloerection
Days 15
There were no further clinical findings on any other study day for this animal
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 49 of 110
Table 28: Individual Body Weight (g) - Males Group
C
LD
MD
HD
Animal No.
Study Day 1
4
8
11
14
1
242
268
285
296
301
2
248
268
290
303
319
3
252
272
293
297
307
4
252
272
297
300
306
5
257
274
302
312
324
6
248
261
287
301
314
7
247
270
288
302
314
8
241
260
281
286
297
9
248
271
291
301
308
10
247
272
287
295
301
11
244
258
275
286
291
12
256
268
284
291
299
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 50 of 110
Table 29: Individual Body Weight (g) - Females Group
C
LD
MD
HD
Animal No.
Study Day 1
4
8
11
14
13
168
176
178
181
180
14
166
168
181
174
186
15
175
179
184
180
189
16
165
165
171
179
182
17
163
164
170
175
177
18
172
188
194
196
198
19
165
174
176
176
176
20
161
164
170
171
179
21
170
179
181
182
185
22
172
175
180
178
184
23
170
178
182
181
182
24
172
182
185
190
195
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 51 of 110
Table 30: Individual Body Weight Gain (g/week) - Males Group C
LD
MD
HD
Animal No. Day 1-4
Day 4-8
Day 8-11
Day 11-14
Day 1-14
17
11
6
59
1
26
2
20
22
12
16
71
3
20
21
3
10
55
4
20
25
3
6
54
5
17
28
10
12
67
6
13
26
15
13
66
7
23
18
14
12
67
8
19
21
5
11
56
9
23
20
10
8
60
10
25
15
9
6
54
11
14
17
11
6
47
12
12
16
7
8
43
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 52 of 110
Table 31: Individual Body Weight Gain (g/week) - Females Group C
LD
MD
HD
Animal No. Day 1-4
Day 4-8
Day 11-14
Day 1-14
3
-1
12
Day 8-11
13
8
2
14
2
13
-7
12
20
15
4
5
-4
9
15
16
0
6
8
3
17
17
2
6
5
2
15
18
16
6
2
1
25
19
9
2
-1
1
11
20
3
6
1
9
18
21
10
2
1
3
16
22
3
5
-2
6
11
23
8
4
-1
2
12
24
10
3
5
5
23
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 53 of 110
Table 32: Individual Food Consumption (g/day) - Males Group
C
LD
MD
HD
Animal No.
Study Day Day 1-7
Day 7-14
Day 1-14
1
22.17
24.87
23.52
2
24.19
28.03
26.11
3
22.34
25.76
24.05
4
24.10
26.73
25.41
5
23.56
28.16
25.86
6
23.37
26.56
24.96
7
25.81
28.81
27.31
8
22.96
24.04
23.50
9
24.27
26.39
25.33
10
24.49
26.60
25.54
11
20.73
23.96
22.34
12
22.44
25.13
23.79
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 54 of 110
Table 33: Individual Food Consumption (g/day) - Females Group
C
LD
MD
HD
Animal No.
Study Day Day 1-8
Day 8-14
Day 1-14
13
13.53
13.94
13.74
14
13.29
15.67
14.48
15
13.90
15.97
14.94
16
14.17
15.50
14.84
17
12.86
14.03
13.44
18
14.96
16.63
15.79
19
12.44
13.19
12.81
20
13.54
15.69
14.61
21
14.46
17.26
15.86
22
13.70
15.24
14.47
23
14.27
16.49
15.38
24
13.86
15.96
14.91
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Table 34: Individual Haematology - Males Group
C
Animal No.
RBC
HB
HCT
PLT
WBC
9
10 /L
g/dL
%
10 /L
109/L
1
8.52
15.8
46.9
1093
8.22
2
8.62
16.3
47.6
1098
4.68
4
8.46
15.8
46.8
1077
5.60
5
8.09
14.9
43.1
1064
5.44
6
8.05
15.6
46.7
1153
10.01
7
8.25
15.7
46.6
1105
4.63
8
8.13
15.8
45.5
839
4.90
9
8.75
16.3
48.0
900
4.80
11
9.28
16.5
48.8
1059
6.84
12
9.31
17.0
50.8
1115
7.05
Units
12
3 LD
MD
10 HD
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 55 of 110
Table 35: Individual Haematology - Females Group
C
Animal No.
RBC
HB
HCT
PLT
WBC
9
10 /L
g/dL
%
10 /L
10 9/L
13
8.36
14.8
44.6
811
3.26
14
7.22
14.2
41.4
947
3.12
15
8.47
15.0
44.7
988
3.08
17
8.54
15.4
46.8
1112
2.59
18
8.86
16.0
46.9
973
2.49
19
8.48
15.6
44.9
869
3.15
20
8.19
14.7
44.3
852
3.42
21
8.23
14.9
43.8
944
3.84
22
8.60
15.0
45.4
864
3.21
23
7.64
14.5
43.7
874
2.97
24
8.81
15.7
46.9
971
3.24
Units
12
16 LD
MD
HD
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 56 of 110
Table 36: Individual Clinical Biochemistry - Males Group
C
LD
MD
Animal No.
ASAT
ALAT
AP
Crea
TP
ALBB
Urea
Chol
Na
K
Units
U/L
U/L
U/L
Âľmol/L
g/L
g/L
mmol/L
mmol/L
mmol/L
mmol/L
1
34.5
19.0
294
30.9
63.2
35.2
7.83
1.08
152
4.22
2
39.3
27.2
n
35.4
60.2
36.1
7.95
0.87
150
3.33
3
32.1
16.6
201
29.7
56.9
32.0
8.76
0.94
148
4.56
4
24.4
19.5
186
29.2
58.3
31.5
8.38
0.78
151
4.65
5
31.3
18.5
229
28.4
57.4
31.6
7.85
0.74
151
4.59
6
32.3
21.1
148
26.0
59.8
33.0
7.66
0.80
151
3.92
7
34.6
18.7
216
26.3
58.6
33.3
7.90
0.67
152
4.00
8
28.8
20.0
135
30.7
58.3
33.4
7.92
0.75
147
3.74
9
27.8
22.9
213
27.5
57.5
33.6
6.85
1.13
154
3.58
11
35.2
24.2
197
29.0
59.5
33.8
8.29
0.69
153
4.25
12
37.0
22.5
168
31.0
59.9
33.8
8.16
0.93
154
4.68
10 HD
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 57 of 110
Table 37: Individual Clinical Biochemistry - Females Group
C
LD
MD
HD
Animal No.
ASAT
ALAT
AP
Crea
TP
ALBB
Urea
Chol
Na
K
U/L
U/L
U/L
Âľmol/L
g/L
g/L
mmol/L
mmol/L
mmol/L
mmol/L
13
44.1
14.9
43
31.6
51.6
32.7
7.12
0.52
144
4.39
14
47.8
14.8
78
34.6
53.1
33.8
7.38
0.58
144
3.38
15
40.8
12.1
81
34.4
50.3
31.5
7.05
0.62
145
3.45
16
36.8
13.5
65
43.0
56.1
34.4
6.65
0.61
144
4.60
17
36.4
15.5
68
26.9
53.5
34.5
6.34
0.46
147
3.59
18
40.7
14.5
71
25.5
51.0
32.2
5.58
0.53
146
3.53
19
30.2
13.8
52
25.2
51.7
33.1
8.73
0.62
142
3.10
20
30.7
12.4
91
29.3
56.0
34.1
6.43
0.54
147
3.31
21
32.3
16.3
85
21.5
55.7
34.6
6.76
0.51
145
3.79
22
41.4
14.7
90
40.0
54.4
31.6
7.96
0.60
143
3.31
23
34.2
15.0
51
31.9
55.6
34.7
8.52
0.66
144
3.39
24
38.0
14.0
80
30.1
55.2
34.0
7.81
0.51
148
3.77
Units
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 58 of 110
Table 38: Individual Macroscopic Findings - Males Group:
C
Animal No.
1 No abnormalities were detected
Animal No.
2 No abnormalities were detected
Animal No.
3 No abnormalities were detected
Group:
LD
Animal No.
4 No abnormalities were detected
Animal No.
5 No abnormalities were detected
Animal No.
6 Organ
Finding
Epididymis
yellow foci
Stomach
discoloured dark
In the remaining organs/tissues no abnormalities were detected
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Group:
MD
Animal No.
7
page 59 of 110
No abnormalities were detected Animal No.
8 No abnormalities were detected
Animal No.
9
No abnormalities were detected Group:
HD
Animal No.
10 Organ Brain
Animal No.
Finding Blood at surface
11 Organ Epididymis
Animal No.
Finding yellow foci
12 Organ Adrenal
Finding Cyst
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 60 of 110
Table 39: Individual Macroscopic Findings - Females Group:
C
Animal No.
13 No abnormalities were detected
Animal No.
14 No abnormalities were detected
Animal No.
15 Organ Stomach
Group:
LD
Animal No.
16
Finding Mucosa thickened
No abnormalities were detected Animal No.
17 No abnormalities were detected
Animal No.
18 No abnormalities were detected
C = Control, LD = Low Dose, MD = Medium Dose, HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
Group:
MD
Animal No.
19 No abnormalities were detected
Animal No.
20 No abnormalities were detected
Animal No.
21 No abnormalities were detected
Group:
HD
Animal No.
22 No abnormalities were detected
Animal No.
23 No abnormalities were detected
Animal No.
24 No abnormalities were detected
page 61 of 110
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 62 of 110
Table 40: Absolute Individual Organ Weights (g) - Males Group
C
LD
MD
HD
Animal No.
Body weight
Brain
Liver
Heart
1
285
1.929
8.74
2
293
2.121
3
284
4 5
Kidneys
Adrenals
Spleen
left
right
total
left
right
total
0.87
1.14
1.15
2.30
0.035
0.032
0.067
0.83
8.62
0.89
1.12
1.12
2.24
0.042
0.040
0.082
0.66
2.087
8.60
0.96
1.14
1.15
2.30
0.036
0.037
0.073
0.73
285
2.022
8.97
1.10
1.13
1.23
2.36
0.042
0.039
0.081
0.72
299
1.968
8.74
1.05
1.03
1.07
2.10
0.036
0.032
0.068
0.67
6
294
1.808
8.55
0.98
1.13
1.17
2.30
0.037
0.040
0.077
0.72
7
290
2.043
9.25
1.01
1.29
1.38
2.67
0.041
0.037
0.079
0.67
8
273
2.085
7.80
0.96
1.07
1.18
2.25
0.033
0.031
0.063
0.80
9
291
1.945
8.51
1.12
1.19
1.19
2.38
0.038
0.028
0.066
0.76
10
284
1.906
13.31
0.93
1.12
1.13
2.24
0.042
0.042
0.084
0.65
11
276
2.038
8.49
0.89
1.16
1.17
2.33
0.035
0.033
0.067
0.61
12
281
2.118
8.87
0.96
1.28
1.19
2.47
0.033
0.035
0.069
0.60
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 63 of 110
Table 40: Absolute Individual Organ Weights (g) - Males (Continued) Group
C
LD
MD
HD
Animal No. Thymus
Testes
Epididymides
left
right
total
left
right
total
Prostate #
1
0.71
1.78
1.73
3.51
0.43
0.46
0.89
1.78
2
0.51
1.67
1.63
3.31
0.46
0.48
0.94
1.84
3
0.69
1.70
1.76
3.46
0.46
0.49
0.95
1.61
4
0.65
1.88
1.77
3.66
0.44
0.43
0.87
1.58
5
0.51
1.50
1.67
3.17
0.40
0.44
0.84
1.54
6
0.48
1.77
1.58
3.35
0.48
0.45
0.93
1.50
7
0.52
1.86
1.86
3.72
0.48
0.46
0.94
1.59
8
0.63
1.63
1.67
3.30
0.40
0.41
0.81
1.49
9
0.53
1.64
1.63
3.27
0.47
0.48
0.95
1.52
10
0.49
1.67
1.69
3.37
0.41
0.48
0.89
1.71
11
0.55
1.57
1.67
3.24
0.50
0.53
1.03
1.99
12
0.58
1.66
1.66
3.32
0.50
0.44
0.94
1.90
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 64 of 110
Table 41: Absolute Individual Organ Weights (g) - Females Group
C
LD
MD
HD
Animal No.
Body weight
Brain
Liver
Heart
13
171
1.759
4.70
14
173
1.811
5.12
15
174
1.775
16
169
17 18
Kidney
Adrenals
left
right
total
left
right
total
0.64
0.76
0.78
1.54
0.032
0.043
0.075
0.60
0.74
0.74
1.48
0.030
0.030
0.060
4.95
0.60
0.68
0.67
1.35
0.036
0.039
0.075
1.786
6.84
0.51
0.70
0.74
1.44
0.043
0.038
0.082
166
1.837
4.62
0.57
0.72
0.73
1.45
0.039
0.034
0.073
178
1.821
4.77
0.64
0.69
0.78
1.46
0.036
0.036
0.071
19
169
1.834
4.33
0.63
0.68
0.73
1.41
0.032
0.032
0.065
20
166
1.801
4.77
0.62
0.57
0.67
1.24
0.042
0.037
0.079
21
176
1.793
5.56
0.59
0.84
0.89
1.73
0.047
0.043
0.091
22
172
1.868
5.45
0.63
0.76
0.79
1.55
0.050
0.037
0.087
23
170
1.675
5.31
0.71
0.81
0.78
1.59
0.043
0.042
0.085
24
180
1.770
5.02
0.57
0.72
0.75
1.47
0.040
0.038
0.077
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 65 of 110
Table 41: Absolute Individual Organ Weights (g) - Females (Continued) Group
C
LD
MD
HD
Animal No. Spleen
Thymus
Ovaries left
right
total
Uterus #
13
0.48
0.35
0.052
0.046
0.097
0.41
14
0.49
0.31
0.045
0.041
0.086
0.61
15
0.43
0.32
0.060
0.060
0.120
0.51
16
0.38
0.27
0.053
0.039
0.092
0.79
17
0.50
0.36
0.036
0.043
0.079
0.86
18
0.43
0.37
0.047
0.044
0.091
0.36
19
0.43
0.26
0.039
0.045
0.083
0.35
20
0.52
0.40
0.041
0.053
0.094
1.03
21
0.46
0.32
0.058
0.052
0.110
0.43
22
0.42
0.28
0.047
0.051
0.098
0.59
23
0.52
0.38
0.045
0.053
0.099
0.68
24
0.48
0.36
0.036
0.049
0.084
0.38
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 66 of 110
Table 42: Relative Individual Organ Weights to Brain Weight (%) - Males Kidneys Group
C
LD
MD
HD
Animal No.
Liver
Heart
1
452.95
2
Adrenals Spleen
left
right
total
left
right
total
45.31
59.29
59.71
119.00
1.825
1.669
3.494
42.80
406.50
41.97
52.76
52.95
105.71
1.971
1.895
3.866
31.20
3
411.99
46.06
54.76
55.32
110.08
1.715
1.768
3.483
34.88
4
443.51
54.53
56.12
60.61
116.73
2.053
1.929
3.981
35.70
5
444.28
53.18
52.11
54.60
106.71
1.845
1.611
3.456
33.94
6
472.93
54.06
62.49
64.62
127.11
2.057
2.212
4.269
39.69
7
453.02
49.28
63.13
67.60
130.72
2.017
1.831
3.847
32.63
8
374.14
46.08
51.30
56.40
107.70
1.569
1.463
3.032
38.20
9
437.69
57.42
61.23
60.93
122.16
1.969
1.434
3.403
39.32
10
698.69
48.63
58.58
59.20
117.78
2.209
2.204
4.413
34.08
11
416.54
43.73
56.81
57.64
114.45
1.698
1.595
3.292
30.05
12
418.97
45.48
60.36
56.04
116.41
1.577
1.671
3.248
28.17
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 67 of 110
Table 42: Relative Individual Organ Weights to Brain Weight (%) - Males (Continued) Group
C
LD
MD
HD
Animal No.
Thymus
1 2
Testes
Epididymides
Prostate #
left
right
total
left
right
total
36.74
92.43
89.70
182.13
22.42
23.81
46.23
92.42
24.10
78.80
77.08
155.88
21.51
22.62
44.13
86.94
3
33.00
81.54
84.31
165.86
22.09
23.57
45.65
77.33
4
32.04
93.11
87.72
180.82
21.71
21.18
42.89
77.94
5
26.02
76.19
84.74
160.92
20.52
22.20
42.72
78.43
6
26.35
97.68
87.52
185.20
26.30
24.99
51.29
82.90
7
25.50
90.98
91.10
182.08
23.53
22.64
46.17
77.65
8
30.00
78.23
80.10
158.33
19.20
19.55
38.75
71.25
9
27.40
84.10
83.82
167.92
24.00
24.88
48.88
78.37
10
25.60
87.83
88.88
176.71
21.67
25.23
46.90
89.63
11
27.23
77.07
82.02
159.09
24.42
25.94
50.36
97.80
12
27.37
78.54
78.14
156.69
23.46
20.89
44.35
89.55
#= including seminal vesicles and coagulating gland
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose; * = including coagulating glands
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 68 of 110
Table 43: Relative Individual Organ Weights to Brain Weight (%) - Females Kidney Group
C
LD
MD
HD
Animal No.
Liver
Heart
13
267.04
14
Adrenals
left
right
total
left
right
total
36.25
43.19
44.18
87.37
1.842
2.422
4.265
282.67
33.12
40.86
41.09
81.96
1.674
1.663
3.336
15
278.58
33.75
38.15
37.99
76.14
2.045
2.203
4.247
16
382.72
28.75
39.21
41.64
80.85
2.430
2.150
4.580
17
251.61
30.80
39.04
39.93
78.98
2.123
1.856
3.979
18
262.10
35.15
37.64
42.76
80.39
1.950
1.966
3.916
19
235.85
34.25
37.14
39.90
77.04
1.756
1.761
3.517
20
264.82
34.49
31.70
37.11
68.80
2.332
2.066
4.398
21
310.17
32.94
47.02
49.55
96.58
2.632
2.414
5.046
22
291.46
33.95
40.47
42.31
82.78
2.687
1.959
4.646
23
316.98
42.23
48.36
46.41
94.77
2.568
2.508
5.076
24
283.89
32.24
40.49
42.40
82.88
2.232
2.119
4.352
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 69 of 110
Table 43: Relative Individual Organ Weights to Brain Weight (%) - Females Group
C
LD
MD
HD
Animal No.
Spleen
Thymus
13
27.14
14
27.20
15
Ovaries
Uterus #
left
right
total
20.08
2.951
2.587
5.538
23.03
17.02
2.502
2.242
4.745
33.92
24.19
18.01
3.386
3.386
6.771
28.64
16
21.29
15.04
2.950
2.195
5.145
44.19
17
26.95
19.46
1.960
2.319
4.279
46.59
18
23.69
20.36
2.581
2.389
4.970
19.78
19
23.17
14.42
2.110
2.432
4.542
18.88
20
28.68
21.99
2.266
2.965
5.231
56.94
21
25.45
17.71
3.245
2.894
6.139
24.18
22
22.60
15.06
2.494
2.730
5.224
31.78
23
30.86
22.51
2.699
3.183
5.882
40.70
24
27.37
20.21
2.006
2.763
4.770
21.35
# including Cervix and Oviduct
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
Report, BSL BIOSERVICE Study No. 112936 Version: Final
page 70 of 110
Table 44: Relative Individual Organ Weights to Body Weight (%) - Males Kidneys Group
C
LD
MD
HD
Animal No.
Brain
Liver
Heart
1
0.677
3.066
2
0.724
3
Adrenals Spleen
left
right
total
left
right
total
0.307
0.401
0.404
0.805
0.0124
0.0113
0.0236
0.290
2.942
0.304
0.382
0.383
0.765
0.0143
0.0137
0.0280
0.226
0.735
3.028
0.339
0.402
0.407
0.809
0.0126
0.0130
0.0256
0.256
4
0.709
3.146
0.387
0.398
0.430
0.828
0.0146
0.0137
0.0282
0.253
5
0.658
2.924
0.350
0.343
0.359
0.702
0.0121
0.0106
0.0227
0.223
6
0.615
2.909
0.332
0.384
0.397
0.782
0.0127
0.0136
0.0263
0.244
7
0.704
3.191
0.347
0.445
0.476
0.921
0.0142
0.0129
0.0271
0.230
8
0.764
2.857
0.352
0.392
0.431
0.822
0.0120
0.0112
0.0232
0.292
9
0.668
2.926
0.384
0.409
0.407
0.817
0.0132
0.0096
0.0227
0.263
10
0.671
4.688
0.326
0.393
0.397
0.790
0.0148
0.0148
0.0296
0.229
11
0.738
3.076
0.323
0.419
0.426
0.845
0.0125
0.0118
0.0243
0.222
12
0.754
3.158
0.343
0.455
0.422
0.877
0.0119
0.0126
0.0245
0.212
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
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Table 44: Relative Individual Organ Weights to Body Weight (%) - Males (Continued) Group
C
LD
MD
HD
Animal No.
Thymus
1 2
Testes
Epididymides
Prostate #
left
right
total
left
right
total
0.249
0.626
0.607
1.233
0.152
0.161
0.313
0.626
0.174
0.570
0.558
1.128
0.156
0.164
0.319
0.629
3
0.243
0.599
0.620
1.219
0.162
0.173
0.336
0.568
4
0.227
0.661
0.622
1.283
0.154
0.150
0.304
0.553
5
0.171
0.501
0.558
1.059
0.135
0.146
0.281
0.516
6
0.162
0.601
0.538
1.139
0.162
0.154
0.315
0.510
7
0.180
0.641
0.642
1.283
0.166
0.160
0.325
0.547
8
0.229
0.597
0.612
1.209
0.147
0.149
0.296
0.544
9
0.183
0.562
0.560
1.123
0.160
0.166
0.327
0.524
10
0.172
0.589
0.596
1.186
0.145
0.169
0.315
0.601
11
0.201
0.569
0.606
1.175
0.180
0.192
0.372
0.722
12
0.206
0.592
0.589
1.181
0.177
0.157
0.334
0.675
# = including seminal vesicles and coagulating gland
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose; * = including seminal vesicles and coagulating gland
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Table 45: Relative Individual Organ Weights to Body Weight (%) - Females Adrenals
Kidney Group
C
LD
MD
HD
Animal No.
Brain
Liver
Heart
13
1.028
2.746
14
1.047
15
1.020
16
left
right
total
left
right
total
0.373
0.444
0.454
0.899
0.0189
0.0249
0.0439
2.958
0.347
0.428
0.430
0.858
0.0175
0.0174
0.0349
2.842
0.344
0.389
0.388
0.777
0.0209
0.0225
0.0433
1.057
4.045
0.304
0.414
0.440
0.854
0.0257
0.0227
0.0484
17
1.107
2.784
0.341
0.432
0.442
0.874
0.0235
0.0205
0.0440
18
1.023
2.681
0.360
0.385
0.437
0.822
0.0199
0.0201
0.0401
19
1.085
2.559
0.372
0.403
0.433
0.836
0.0191
0.0191
0.0382
20
1.085
2.873
0.374
0.344
0.403
0.746
0.0253
0.0224
0.0477
21
1.019
3.161
0.336
0.479
0.505
0.984
0.0268
0.0246
0.0514
22
1.086
3.166
0.369
0.440
0.460
0.899
0.0292
0.0213
0.0505
23
0.985
3.122
0.416
0.476
0.457
0.934
0.0253
0.0247
0.0500
24
0.983
2.791
0.317
0.398
0.417
0.815
0.0219
0.0208
0.0428
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
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Table 45: Relative Individual Organ Weights to Body Weight (%) - Females (Continued) Group
C
LD
MD
HD
Animal No.
Spleen
Thymus
13
0.279
14
0.285
15
Ovaries
Uterus #
left
right
total
0.206
0.0304
0.0266
0.0570
0.237
0.178
0.0262
0.0235
0.0497
0.355
0.247
0.184
0.0345
0.0345
0.0691
0.292
16
0.225
0.159
0.0312
0.0232
0.0544
0.467
17
0.298
0.215
0.0217
0.0257
0.0473
0.516
18
0.242
0.208
0.0264
0.0244
0.0508
0.202
19
0.251
0.156
0.0229
0.0264
0.0493
0.205
20
0.311
0.239
0.0246
0.0322
0.0567
0.618
21
0.259
0.180
0.0331
0.0295
0.0626
0.246
22
0.246
0.164
0.0271
0.0297
0.0567
0.345
23
0.304
0.222
0.0266
0.0314
0.0579
0.401
24
0.269
0.199
0.0197
0.0272
0.0469
0.210
C = Control; LD = Low Dose; MD = Medium Dose; HD = High Dose
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13.2. Appendix 2: Histopathology Report
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