Aus dem Institut für Tierschutz, Tierverhalten und Versuchstierkunde des Fachbereichs Veterinärmedizin der Freien Universität Berlin Refinement strategies in breeding and keeping of laboratory mice Inaugural-Dissertation zur Erlangung des Grades eines Doktors der Veterinärmedizin an der Freien Universität Berlin vorgelegt von Charlotte Sophie Leidinger Tierärztin aus Homburg Berlin 2017 Journal-Nr.: 4016 Gedruckt mit Genehmigung des Fachbereichs Veterinärmedizin der Freien Universität Berlin Dekan: Univ.-Prof. Dr. Jürgen Zentek Erster Gutachter: Univ.-Prof. Dr. Christa Thöne-Reineke Zweiter Gutachter: Prof. Dr. Ulrich Förstermann Dritter Gutachter: PD Dr. Bettina Bert Deskriptoren (nach CAB-Thesaurus): Mus musculus; mice; laboratory mammals; animal welfare; breeding; training of animals; positive reinforcement (MeSH); handling; anthrozoology Tag der Promotion: 15.11.2017 Bibliografische Information der Deutschen Nationalbibliothek Die Deutsche Nationalbibliothek verzeichnet diese Publikation in der Deutschen Nationalbibliografie; detaillierte bibliografische Daten sind im Internet über <http://dnb.ddb.de> abrufbar. ISBN: 978-3-86387-872-6 Zugl.: Berlin, Freie Univ., Diss., 2017 Dissertation, Freie Universität Berlin D 188 Dieses Werk ist urheberrechtlich geschützt. Alle Rechte, auch die der Übersetzung, des Nachdruckes und der Vervielfältigung des Buches, oder Teilen daraus, vorbehalten. Kein Teil des Werkes darf ohne schriftliche Genehmigung des Verlages in irgendeiner Form reproduziert oder unter Verwendung elektronischer Systeme verarbeitet, vervielfältigt oder verbreitet werden. Die Wiedergabe von Gebrauchsnamen, Warenbezeichnungen, usw. in diesem Werk berechtigt auch ohne besondere Kennzeichnung nicht zu der Annahme, dass solche Namen im Sinne der Warenzeichen- und Markenschutz-Gesetzgebung als frei zu betrachten wären und daher von jedermann benutzt werden dürfen. This document is protected by copyright law. No part of this document may be reproduced in any form by any means without prior written authorization of the publisher. Alle Rechte vorbehalten | all rights reserved Mensch und Buch Verlag 2017 Choriner Str. 85 - 10119 Berlin [email protected] – www.menschundbuch.de Dedicated to The Foundling Fox CONTENT 1 INTRODUCTION ...................................................................................................................... 1 1.1 THE ECOLOGY OF WILD AND LABORATORY MUS MUSCULUS ....................................... 2 1.1.1 BEHAVIORAL CHARACTERISTICS OF MUS MUSCULUS .......................................... 3 1.1.2 ENVIRONMENTAL FACTORS FACED BY THE WILD MUS MUSCULUS .................... 3 1.1.3 ENVIRONMENTAL FACTORS IN KEEPING OF THE LABORATORY MOUSE ........... 4 1.1.4 REPRODUCTION AND BREEDING OF MUS MUSCULUS .......................................... 7 1.2 ANIMAL TRAINING ............................................................................................................... 11 1.2.1 THE HISTORY OF ANIMAL TRAINING ........................................................................ 11 1.2.2 LEGAL REGULATION OF LABORATORY ANIMAL TRAINING .................................. 11 1.2.3 METHODS OF ANIMAL TRAINING .............................................................................. 12 1.2.4 EFFECT OF TRAINING ON ANIMAL WELFARE ......................................................... 13 1.2.5 CLICKER TRAINING – A SPECIAL FORM OF POSITIVE REINFORCEMENT TRAINING ...................................................................................................................... 15 1.3 OVERALL AIM OF THE STUDY ............................................................................................ 15 1.3.1 PARTICULAR AIM OF THE BREEDING STUDY ......................................................... 16 1.3.2 PARTICULAR AIM OF THE CLICKER TRAINING STUDY .......................................... 16 2 MATERIAL AND METHODS ................................................................................................. 17 2.1 ETHICS STATEMENT ........................................................................................................... 17 2.2 MATERIAL ............................................................................................................................. 17 2.2.3 ANIMALS AND HOUSING ............................................................................................ 18 2.3 EXPERIMENTAL DESIGN .................................................................................................... 19 2.3.1 TREATMENT PROTOCOL FOR THE BREEDING STUDY ......................................... 19 2.3.2 TREATMENT PROTOCOL FOR THE CLICKER TRAINING STUDY .......................... 20 2.3.3 BEHAVIOR TESTS........................................................................................................ 27 2.4 STATISTICAL ANALYSIS ..................................................................................................... 29 3 RESULTS .............................................................................................................................. 31 3.1 ANALYSIS OF THE BREEDING STUDY .............................................................................. 31 3.3 ANALYSIS OF THE CLICKER TRAINING STUDY ................................................................ 38 4 DISCUSSION ......................................................................................................................... 43 4.1 DISCUSSION OF THE BREEDING STUDY .......................................................................... 43 4.2 DISCUSSION OF THE CLICKER TRAINING STUDY ........................................................... 46 5 CONCLUSION ....................................................................................................................... 49 I 6 SUMMARY – REFINEMENT STRATEGIES IN BREEDING AND KEEPING OF LABORATORY MICE ............................................................................................................ 50 7 ZUSAMMENFASSUNG – REFINEMENTSTRATEGIEN IN DER ZUCHT UND HALTUNG VON LABORMÄUSEN .......................................................................................................... 52 8 REFERENCES ....................................................................................................................... 55 9 LIST OF OWN PUBLICATIONS ........................................................................................... 67 10 ACKNOWLEDGMENTS ........................................................................................................ 69 11 DECLARATION OF ORGINALITY ........................................................................................ 70 II LIST OF FIGURES FIGURE 1 C57BL/6JRJ MOUSE INTERACTING WITH NEST BUILDING MATERIAL ..................................................... 5 FIGURE 2 C57BL/6JRJ DAM WITH 5‐DAY‐OLD LITTER ............................................................................................. 8 FIGURE 3 TIMELINE OF THE BREEDING STUDY ..................................................................................................... 19 FIGURE 4 EXAMPLE OF A PETRI DISH WITH DIFFERENT FOOD REWARDS (LEIDINGER ET AL. 2017A) .................. 21 FIGURE 5 TIMELINE OF THE CLICKER TRAINING STUDY ........................................................................................ 21 FIGURE 6 “POSITION OF MOUSE TUNNEL” (LEIDINGER ET AL. 2017A) ................................................................ 23 FIGURE 7 "PRESENTING THE REWARD" (LEIDINGER ET AL. 2017A) ...................................................................... 24 FIGURE 8 “TRAINING SUCCESS” (LEIDINGER ET AL. 2017A) .................................................................................. 25 FIGURE 9 “REWARDING” (LEIDINGER ET AL. 2017A) ............................................................................................ 25 FIGURE 10 "ALTERNATING TARGET STICK POSITIONS IN THE SECOND WEEK OF TRAINING" (LEIDINGER ET AL. 2017A) .................................................................................................................. 26 FIGURE 11 "TRAINING SUCCESS" (LEIDINGER ET AL. 2017A) ................................................................................ 26 FIGURE 12 SETUP OF THE MORRIS WATER MAZE IN THE MBU ............................................................................ 27 FIGURE 13 LITTER SIZE AND PUP LOSS ON P1.. ..................................................................................................... 31 FIGURE 14 LITTER SIZE AND PUP LOSS ON P24. .................................................................................................... 32 FIGURE 15 PROPORTION OF LITTERS THAT WERE LOST COMPLETELY IN RELATION TO LITTERS WHERE ONLY SINGLE MICE DIED. .................................................................................................................... 32 FIGURE 16 PUP WEIGHT ON P1. ........................................................................................................................... 33 FIGURE 17 PUP WEIGHT ON P24. ......................................................................................................................... 34 FIGURE 18 VISUAL EXAMINATIONS ON P1 ........................................................................................................... 35 FIGURE 19 VISUAL EXAMINATIONS ON P7. .......................................................................................................... 35 FIGURE