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Report, BSL BIOSERVICE Study No. 112936 Version: Final

1.

page 2 of 110

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

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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.


Report, BSL BIOSERVICE Study No. 112936 Version: Final

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.


Report, BSL BIOSERVICE Study No. 112936 Version: Final

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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.


Report, BSL BIOSERVICE Study No. 112936 Version: Final

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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:

page 25 of 110

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

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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

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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


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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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Oral Toxiteit final_OECD_112936_V11_00