Tierarztl Prax Ausg G Grosstiere Nutztiere 2019; 47(04): 256-262
DOI: 10.1055/a-0884-8553
Kasuistik
© Georg Thieme Verlag KG Stuttgart · New York

Geburt eines Fohlens nach Besamung mit tiefgefrorenen Nebenhodenspermien und zeitgleicher intrauteriner Verabreichung von homologem Seminalplasma

Birth of a foal following insemination with frozen-thawed epididymal sperm and simultaneous intrauterine delivery of homologous seminal plasma
Stefanie Neuhauser
Pferdezentrum Bad Saarow, Abteilung für Reproduktionsmedizin beim Pferd, Fachbereich Veterinärmedizin, Freie Universität Berlin
,
Patricia Gösele
Pferdezentrum Bad Saarow, Abteilung für Reproduktionsmedizin beim Pferd, Fachbereich Veterinärmedizin, Freie Universität Berlin
,
Johannes Handler
Pferdezentrum Bad Saarow, Abteilung für Reproduktionsmedizin beim Pferd, Fachbereich Veterinärmedizin, Freie Universität Berlin
› Author Affiliations
Further Information

Publication History

10 August 2018

22 November 2018

Publication Date:
21 August 2019 (online)

Zusammenfassung

Die Tiefgefrierkonservierung von Nebenhodenspermien stellt eine gute Möglichkeit zur Erhaltung der Genreserve wertvoller Vatertiere dar, wenn ein plötzlicher Unfall eine herkömmliche Samengewinnung unmöglich macht. Im vorliegenden Fall wird von der Geburt eines gesunden Fohlens berichtet, das aus der Besamung mit Nebenhodenspermien entstand. Die Hoden und Nebenhoden wurden in Allgemeinanästhesie entfernt und gekühlt über Nacht in das Labor verbracht. Durch retrograde Spülung der Nebenhodenschwänze konnten 17,79 × 109 Spermien konserviert werden. Eine fruchtbare Stute wurde 1 Jahr später mit tiefgefrorenen Nebenhodenspermien tief intrauterin besamt. Im 1. Zyklus wurde die Stute nicht tragend. Im 2. Zyklus wurde homologes Seminalplasma zusammen mit den Nebenhodenspermien in den Uterus verbracht und 14 Tage nach der Ovulation konnte eine Trächtigkeit festgestellt werden. Nach einer Graviditätsdauer von 334 Tagen wurde ein gesundes Hengstfohlen geboren. Die Vorbereitung von Hoden und Nebenhoden nach einem Unfall oder Todesfall für die weitere Bearbeitung im Labor und die einzelnen Arbeitsschritte für die Aufbereitung von Nebenhodenspermien werden beschrieben.

Abstract

Cryopreservation of epididymal sperm allows final preservation of the gene reserve from valuable sires in case of unexpected injury terminating the breeding career. This case report describes the birth of a healthy foal following insemination with frozen-thawed epididymal sperm. The testes and epididymides were removed under general anaesthesia and sent cooled to the laboratory overnight. The cauda epididymidis was dissected and 17.79 × 109 sperm were harvested by a retrograde flush technique. A fertile mare was inseminated 1 year later with frozen-thawed epididymal sperm. Sperm were deposited into the tip of the uterine horn ipsilateral to the ovulation site. The mare did not become pregnant after the 1st cycle. In the 2nd breeding cycle, additional homologous seminal plasma was delivered into the uterus at the time of insemination and the mare was diagnosed as pregnant 14 days post ovulation. A healthy colt was born after 334 days of gestation. The method for preparing gonads for transportation to an appropriate laboratory is described for veterinarians and the different steps of semen collection and preservation are presented.

 
  • Literatur

  • 1 Al-Essawe EM, Wallgren M, Wulf M. et al. Seminal plasma influences the fertilizing potential of cryopreserved stallion sperm. Theriogenology 2018; 115: 99-107
  • 2 Alvarenga MA, Papa FO, Landim-Alvarenga FC. et al. Amides as cryoprotectants for freezing stallion semen: a review. Anim Reprod Sci 2005; 89: 105-113
  • 3 Amann RP, Pickett BW. Principles of cryopreservation and a review of cryopreservation of stallion spermatozoa. J Equine Vet Sci 1987; 7: 145-173
  • 4 Barker CAV, Gandier JCC. Pregnancy in a mare resulting from frozen epididymal spermatozoa. Can J Comp Med 1957; 21: 47-51
  • 5 Barrier-Battut I, Bonnet C, Giraudo A. et al. Removal of seminal plasma enhances membrane stability on fresh and cooled stallion spermatozoa. Reprod Dom Anim 2013; 48: 64-71
  • 6 Batista M, Vilar J, Rosario I. et al. Influence of different anaesthetic protocols over the sperm quality on the fresh, chilled (4° C) and frozen-thawed epididymal sperm samples in domestic dogs. Reprod Dom Anim 2016; 51: 758-765
  • 7 Bruemmer JE. Collection and freezing of epididymal stallion sperm. Vet Clin Equine 2006; 22: 677-682
  • 8 De Oliveira RA, Budik S, Aurich C. Influence of partial or total replacement of glycerol by alternative cryoprotectants in Ghent freezing extender on post-thaw sperm quality in stallions. Reprod Dom Anim 2017; 52: 715-721
  • 9 Fernández-Santos MR, Soler AJ, Ramón M. et al. Effect of post-mortem time on post-thaw characteristics of Spanish ibex (Capra pyrenaica) spermatozoa. Anim Reprod Sci 2011; 129: 56-66
  • 10 Gloria A, Carluccio A, Petrizzi L. et al. Characteristics of frozen epididymal spermatozoa from stallions that died 12 to 36 hours after colic surgery. Theriogenology 2016; 85: 345-350
  • 11 Guimarães T, Lopes G, Ferreira P. et al. Characteristics of stallion epididymal spermatozoa at collection and effect of two refrigeration protocols on the quality of the frozen/thawed sperm cells. Anim Reprod Sci 2012; 136: 85-89
  • 12 Handler J, Neuhauser S. Influence of seminal plasma on pregnancy rates of frozen-thawed epididymal sperm. Reprod Dom Anim 2014; 49 (Suppl. 03) 71 (Abstract)
  • 13 Hayden SS, Blanchard TL, Brinsko SP. et al. Pregnancy rates in mares inseminated with 0.5 or 1 million sperm using hysteroscopic or transrectally guided deep-horn insemination techniques. Theriogenology 2012; 78: 914-920
  • 14 Heise A, Kähn W, Volkmann DH. et al. Influence of seminal plasma on fertility of fresh and frozen-thawed stallion epididymal spermatozoa. Anim Reprod Sci 2010; 118: 48-53
  • 15 Heise A, Thompson PN, Gerber D. Influence of seminal plasma on fresh and post-thaw parameters of stallion epididymal spermatozoa. Anim Reprod Sci 2011; 123: 192-201
  • 16 Jiménez E, Pérez-Marín CC, Millán Y. et al. Influence of anaesthetic drugs on the epididymal sperm quality in domestic cats. Anim Reprod Sci 2011; 123: 265-269
  • 17 Katila T. Post-mating inflammatory responses of the uterus. Reprod Dom Anim 2012; 47: 31-41
  • 18 Klug E, Sieme H, Peters E. Hygienegrundlagen zum Einsatz von Hengsten in der instrumentellen Samenübertragung. Tierärztl Prax Ausg G 1998; 26: 218-224
  • 19 Macías García B, Ortega Ferrusola C, Aparicio IM. et al. Toxicity of glycerol for the stallion spermatozoa: effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential. Theriogenology 2012; 77: 1280-1289
  • 20 Monteiro GA, Guasti PN, Rocha AS. et al. Effect of storage time and temperature of equine epididymis on the viability, motion parameters, and freezability of epididymal sperm. J Equine Vet Sci 2013; 33: 169-173
  • 21 Monteiro GA, Papa FO, Zahn FS. et al. Cryopreservation and fertility of ejaculated and epididymal stallion sperm. Anim Reprod Sci 2011; 127: 197-201
  • 22 Moore AI, Squires EL, Graham JK. Effect of seminal plasma on the cryopreservation of equine spermatozoa. Theriogenology 2005; 63: 2372-2381
  • 23 Morrell JM, Pihl J, Dalin A-M. et al. Restoration of seminal plasma to stallion spermatozoa selected by colloid centrifugation increases sperm progressive motility but is detrimental to chromatin integrity. Theriogenology 2012; 78: 345-352
  • 24 Morris L, Tiplady C, Allen WR. The in vivo fertility of cauda epididymal spermatozoa in the horse. Theriogenology 2002; 58: 643-646
  • 25 Morris LHA, Tiplady C, Allen WR. Pregnancy rates in mares after a single fixed time hysteroscopic insemination of low numbers of frozen-thawed spermatozoa onto the uterotubal junction. Equine Vet J 2003; 35: 197-201
  • 26 Neuhauser S, Dörfel S, Handler J. Dose-dependent effects of homologous seminal plasma on motility and kinematic characteristics of post-thaw stallion epididymal spermatozoa. Andrology 2015; 3: 536-543
  • 27 Neuhauser S, Gösele P, Handler J. The effect of four different semen extenders on the motility of stallion epididymal sperm. J Equine Vet Sci 2018; 62: 8-12
  • 28 Neuhauser S, Rheinfeld S, Handler J. Motility of fresh and frozen-thawed stallion sperm from three segments of the epididymal cauda and the effect of post-thaw seminal plasma addition on motility. J Equine Vet Sci 2013; 33: 942-949
  • 29 Olaciregui M, Gil L, Montón A. et al. Cryopreservation of epididymal stallion sperm. Cryobiology 2014; 68: 91-95
  • 30 Papa FO, Melo CM, Fioratti EG. et al. Freezing of stallion epididymal sperm. Anim Reprod Sci 2008; 107: 293-301
  • 31 Rigby S, Derczo S, Brinsko S. et al. Oviductal sperm numbers following proximal uterine horn or uterine body insemination. 46th Proc AAEP 2000; 332-334
  • 32 Schulman ML, Gerber D, Nurton J. et al. Effects of halothane anaesthesia on the cryopreservation of epididymal spermatozoa in pony stallions. Equine Vet J 2003; 35: 93-95
  • 33 Slott VL, Linder RE, Dyer CJ. Method of euthanasia does not affect sperm motility in the laboratory rat. Reprod Toxicol 1994; 8: 371-374
  • 34 Smits K, Hoogewijs M, Woelders H. et al. Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids. Reprod Dom Anim 2012; 47 (Suppl. 04) 239-248
  • 35 Sostaric E, Kraan H, Stout TAE. Role of seminal plasma in the attainment of fertilizing capacity by stallion epididymal sperm. Anim Reprod Sci 2010; 121 S: S184-185
  • 36 Squires EL, Keith SL, Graham JK. Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology 2004; 62: 1056-1065
  • 37 Stout TAE, Morris LHA, Li X. et al. The effect of seminal plasma on the motility and cryopreservability of horse epididymal sperm. Havemeyer Foundation Monograph Series 2000; 1: 5-6
  • 38 Töpfer-Petersen E, Ekhlasi-Hundrieser M, Kirchhoff C. et al. The role of stallion seminal proteins in fertilisation. Anim Reprod Sci 2005; 89: 159-170
  • 39 Troedsson MHT, Alghamdi AS, Mattisen J. Equine seminal plasma protects the fertility of spermatozoa in an inflamed uterine environment. Theriogenology 2002; 58: 453-456
  • 40 Vieira LA, Gadea J, García-Vázquez FA. et al. Equine spermatozoa stored in the epididymis for up to 96 h at 4° C can be successfully cryopreserved and maintain their fertilization capacity. Anim Reprod Sci 2013; 136: 280-288
  • 41 Watson PF. Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod Fertil Dev 1995; 7: 871-891