Szent István University Postgraduate School of Veterinary Science Sexual endocrine diagnostic and treatment methods in the domestic ferret ( Mustela putorius furo ) Ph.D. dissertation Written by: Dr. Angella Proháczik Supervisor: Prof. Gyula Huszenicza 2009 1 Szent István Egyetem Állatorvos-tudományi Doktori Iskola Témavezető: ..................................... Dr. Huszenicza Gyula egyetemi tanár, DSc Szent István Egyetem Állatorvostudományi Kar Szülészeti és Szaporodásbiológiai Tanszék és Klinika Témabizottsági tagok: ..................................... Dr. Hornung Erzsébet int. vez. egyetemi docens, CSc Szent István Egyetem Állatorvos-tudományi Kar Biológiai Intézet ..................................... Dr. Rudas Péter tv. egyetemi tanár, DSc Szent István Egyetem Állatorvos-tudományi Kar Élettani és Biokémiai Tanszék Az értekezés a Dr. Huszenicza Gyula témavezető elnökletével Budapesten 2009. augusztus 28. án tartott munkahelyi vita nyomán nyerte el végleges formáját. Készült 8 példányban. Ez a ___. sz. példány. …………………………………. dr. Proháczik Angella 2 List of content ABBREVIATIONS AND ACRONYMS...................................................................................................5 SUMMARY.................................................................................................................................................6 ÖSSZEFOGLALÁS ...................................................................................................................................8 1. INTRODUCTION AND THE AIMS OF THE STUDIES.........................................................10 2. REVIEW OF LITERATURE ......................................................................................................12 2.1. TAXONOMY , DOMESTICATION AND CURRENT STATUS OF FERRETS........................................12 2.2. REPRODUCTION IN THE DOMESTIC FERRET .............................................................................15 2.2.1. Influence of photoperiod, puberty and breeding season.................................................15 2.2.2. Anoestrus, oestrus cycle and ovulation...........................................................................18 2.3. HYPEROESTROGENISM IN THE DOMESTIC FERRET ..................................................................24 2.3.1. Prolonged oestrus...........................................................................................................25 2.3.2. Ovarian remnant syndrome ............................................................................................28 2.3.3. Nodular hyperplasia of adrenocortex.............................................................................30 2.3.4. Unusual cause of the elevated sexual steroid levels in neutered ferrets.........................35 2.4. CORPUS LUTEUM FUNCTION AND MAINTENANCE OF PREGNANCY ..........................................37 2.5. PSEUDOPREGNANCY .................................................................................................................40 2.6. GESTATION , PARTURITION , LACTATION AND RE -BREEDING ...................................................41 2.6.1. Gestation.........................................................................................................................41 2.6.2. Pregnancy toxemia .........................................................................................................42 2.6.3. Parturition ......................................................................................................................43 2.6.4. Lactation and interaction between milk production and ovarian function.....................44 2.6.5. Re-breeding after nursing, abortion or early weaning...................................................46 2.7. LUTEINISATION WITHOUT OVULATION ....................................................................................47 2.8. CURRENT POSSIBILITIES FOR CONTROLLING REPRODUCTION IN FERRETS ............................49 2.8.1. Progestins .......................................................................................................................49 2.8.2. hCG and GnRH ..............................................................................................................51 2.8.3. Long-acting GnRH agonists ...........................................................................................51 2.8.4. Spaying ...........................................................................................................................54 2.9. FECAL P4 MONITORING IN MUSTELIDS .....................................................................................55 3. MATERIALS AND METHODS..................................................................................................57 3.1. COLLECTION , PREPARATION , AND STORAGE OF SAMPLES ..........................................................57 3.2. OESTRUS DETECTION AND MATING SYSTEM ................................................................................58 3.3. OPERATIVE INTERVENTIONS IN GENITALS (E XP . 1A, 2B, 4 AND 5) ...............................................58 3.4. LABORATORY PROCEDURES ..........................................................................................................58 3.4.1. Fecal steroid extraction and P 4-met determination (Exp. 1a, 1b, 1c, 2a, 2b and 3) .............58 3.4.2. Measurements of P 4 in plasma (Exp. 1a)...............................................................................59 3.4.3. Assaying E 2 in plasma (Exp. 4) .............................................................................................60 3.4.4. Measurements of other hormones and metabolites in plasma (Exp. 5).................................60 3.4.5. Histology of surgically removed ovaries (Exp. 2b) ...............................................................61 3.5. STATISTICS ....................................................................................................................................61 4. STUDIES........................................................................................................................................63 4.1. BIOLOGICAL VALIDATION OF FECAL P4-MET ELISA IN FERRETS (E XP . 1A, 1B AND 1C) 63 4.1.1. Design and results.................................................................................................................63 4.1.2. Discussion and conclusions...................................................................................................64 4.2. FECAL P4-MET PATTERN AND REPRODUCTIVE FEATURES OF JILLS KEPT UNDER KENNEL CONDITIONS (E XP . 2A AND 2B).............................................................................................................65 4.2.1. Materials and methods ..........................................................................................................66 4.2.2. Results ...................................................................................................................................68 4.2.2.1. Fecal P 4-met pattern and oestrus in lactating and weaned jills (Exp 2a and 2b) ..............68 4.2.2.2. Ovarian histology in lactating and non-lactating postpartum jills (Exp. 2b).....................71 4.2.2.3 Further experiences produced by the continuous monitoring of individual P 4-met pattern throughout the breeding season ......................................................................................................74 4.2.3. Discussion and conclusions...................................................................................................76 4.3. COMPARATIVE STUDY ON SUPPRESSION OF OVARIAN ACTIVITY IN FERRETS (E XP . 3) .....79 3 4.3.1. Material and methods............................................................................................................79 4.3.1.1. Animals...............................................................................................................................79 4.3.1.2. Design and treatments........................................................................................................80 4.3.1.3. Assessment of reproductive and clinical response .............................................................80 4.3.1.4. Statistics and evaluation of the results ...............................................................................81 4.3.2. Results ...................................................................................................................................81 4.3.3. Discussion and conclusions...................................................................................................88 4.4. EXPERIENCE WITH A NON -TRADITIONAL TREATMENT OF HYPEROESTROGENISM IN FERRETS (E XP . 4) ..................................................................................................................................91 4.4.1. Material and methods............................................................................................................91 4.4.1.1. Animals and design.............................................................................................................91 4.4.1.2. Handling of blood samples. Statistical procedures ............................................................94 4.4.2. Results ...................................................................................................................................94 4.4.3. Discussion and conclusions...................................................................................................95