Insemination with frozen dog semen based on a new insemination technique.
ABSTRACT Inhalt: (Insemination mit Gefriersperma beim Hund mit Hilfe einer neuen Besamungstechnik).In diesem Versuch sind 11 Hündinnen mit tiefgefrorenem Sperma besamt worden.Die spermienreiche Ejakulatsfraktion wurde unmittelbar nach der Samenentnahme mit einer Tris-Fructose-Zitronensüure-Lösung, die 8 Vol. % Glyzerin und 20 Vol. % Eidotter enthielt, etwa 1: 4 verdünnt. Wührend einer 3stündigen Equilibrierungszeit wurde der Samen auf + 5 °C heruntergekühlt, and in PVC-Röhrchen im Stickstoffdampf eingefroren.Die Inseminationsdosen, die ungefähr 150 × 106 Spermien enthielten und 3 Wochen bis 1 1/2 Jahre gelagert waren, wurden unmittelbar nach dem Auftauen für 6,5 Sek. im Wasserbad von + 75 °C, durch den Cervicalkanal intrauterin deponiert. Es wurde meistens zweimal mit etwa 48stündigen Intervallen inseminiert. Von den 11 Hündinnen konzipierten 10 und brachten von ein bis sieben Junge. Die höchste Zahl der lebendigen Welpen in einem Wurf war sechs.Contents:Insemination with frozen dog semen was performed in a small trial including 11 bitches. The semen was diluted about 1: 4 with Tris-fructose-citric acid extender containing 8 % (v/v) glycerol and 20 % (v/v) egg yolk, equilibrated for 3 hrs and frozen in P.V.C.-straws by use of N2-vapour. The insemination doses being stored in liquid N2 from 3 weeks to 1 1/2 year and containing about 150 × 106 spermätozoa, was deposited in the uterus via the cervical canal immediately after thawing at 75 °C for 6,5 sec. By this procedure conception was obtained in 10 of the 11 bitches. The litter size ranged from one to seven puppies, the highest number of living puppies being six.
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ABSTRACT: Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.Asian Australasian Journal of Animal Sciences 06/2014; 27(6):791-6. · 0.64 Impact Factor
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ABSTRACT: The dog has been used in medical research as a model for humans, but the lack of appreciation of the differences especially in reproductive pattern and hormonal effects and sensitivity has led to some classic misconceptions for instance regarding the tumorigenic effects of progestagens on mammary glands. In contrast, the dog has proved to be a very useful model in studies on human prostatic functions. The domestic dog is also more and more used as a model in research aiming at the preservation of the many species of wild canids that are threatened by extinction; projects popularly referred to as "The Frozen Zoo". During the last 10-15 years our knowledge of canine reproduction has made major progress. This chapter aims at summarising the basic reproductive physiology of the dog, including the latest discoveries within this field, and also to give an update on the applications of new reproductive technology in this species. Reproductive endocrinology The reproductive events, both in the male and the female dog, are orchestrated from the hypothalamus which in response to some as yet partly unknown stimuli produces and releases the gonadotrophin-releasing hormone (GnRH), which, in turn, influences the pituitary gland to secrete follicle stimulating hormone (FSH) and luteinizing hormone (LH). These gonadotrophic hormones induce ovarian follicular development and ovulation in the bitch and testicular development, androgen production and spermatogenesis in the male. The hypothalamic-pituitary-gonadal axis is regulated via intricate feedback mechanisms (Fig 1a, b) whereby the gonadal hormones having reached a certain concentration via negative feedback downregulate further release of GnRH, and thus FSH and LH. Reproduction in the Male The reproductive organs of the male dog consist of the testicles with epididymides and the vas deferens, the prostate gland, the urethra and the penis. The testicle contains the seminiferous tubules, producing spermatozoa, and the interstitium with Leydig cells which produce steroids, particularly testosterone, in the sexually mature individual. The epididymis consists of a single long duct in which the spermatozoa during passage undergo maturational changes and obtain the capacity for motility. The distal part of the epididymis, the cauda, is the storage site for the matured spermatozoa. Prostatic fluid constitutes the major portion of the ejaculate, and contains several enzymes, cholesterol and lactate. The penis consists of a pelvic part, and01/2001;
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ABSTRACT: The objective of this retrospective clinical study was to compare pregnancy rates obtained after the use of endoscopic-assisted transcervical catheterization (EIU) or laparotomy (SIU) for insemination of frozen-thawed dog semen. Healthy bitches from various breeds were inseminated with semen from multiple donors processed by different freezing centers. Data from 118 inseminations (78 EIU and 40 SIU) performed between 2009 and 2011 were analyzed. Insemination timing was based on vaginal cytology, serum progesterone concentrations, and vaginoscopy. A ureterorenoscope and a CH-5 Transcervical insemination catheter were used for EIU; 28 of the bitches in this group were inseminated twice with the second insemination less than 12 hours after the first. The numbers of live morphologically normal sperm (LMNS) were determined to characterize insemination doses. Overall, pregnancy rate was greater (P < 0.05) in the EIU group (65%) than in the SIU group (45%). Pregnancy rates were greater (P ≤ 0.06) when more than 100 × 10(6) LMNS were inseminated regardless of insemination method; the greatest pregnancy rate was observed in the EIU group when this insemination dose was used (38/49; 78%). There was no significant difference in pregnancy rate whether one (69%) or two inseminations (64%) were performed in the EIU group. Complications in the SIU group included anesthetic-induced bradycardia during surgery, significant postsurgery pain, seroma formation over the abdominal incision, and delayed wound healing. No complications were noted during or after insemination in the EIU group. In conclusion, these results support the use of EIU as a noninvasive alternative to laparotomy for insemination of frozen-thawed dog semen. In addition, use of more than 100 × 10(6) LMNS is also recommended for insemination.Theriogenology 06/2014; · 2.08 Impact Factor