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: Developmental competence and quality of in vitro produced embryos has been demonstrated to be lower than in vivo derived embryos. This study aimed specifically to determine the effects of in vitro culture of feline embryos using various culture densities on developmental competence and expression of stress- and apoptotic-related genes in terms of heat shock protein 70 (HSP70) and apoptotic-related (BAX and BCL-2) gene expressions. In experiment 1, we characterized the inducible form of a feline-specific HSP70 mRNA sequence, as it has not been previously reported. The primers for feline HSP70 mRNA were synthesized and tested on heat-treated cat fibroblasts. In experiment 2, feline embryos were cultured at different culture densities (embryo:culture volume; 1:1.25, 1:5 and 1:20). The developmental competence was determined along with HSP70, BAX and BCL-2 transcript abundances using quantitative RT-PCR. In vivo derived embryos were used as a control group. A partial cat HSP70 mRNA sequence (190 bp) was characterized and exhibited high nucleotide identity (93 to 96%) with other species. Cleaved embryos cultured at high density (1:1.25) developed to blastocysts at a lower rate than those generated from lower densities. Irrespective of the culture densities used, in vitro cultured blastocysts showed increased levels of HSP70 and BAX transcripts compared with in vivo counterparts. Blastocysts derived from the highest culture density (1:1.25) showed higher levels of upregulation of HSP70 and BAX transcripts than those cultured at lower culture densities (1:5 and 1:20). In conclusion, increased levels of pro-apoptotic (BAX) and stress-response (HSP70) transcripts correlated with developmental incompetence of embryos cultured at high embryonic density, indicating that stress accumulated during in vitro embryo culture affected the fate for embryo development and quality.Journal of Reproduction and Development 01/2013; · 1.76 Impact Factor
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ABSTRACT: (1) To report whelping rates and litter sizes following coeliotomy-assisted intrauterine inseminations (CAII) performed commercially and (2) to identify factors that may influence these outcomes. Retrospective single cohort observational study. All oestrous cycles in bitches that presented to the study hospital for CAII between 1 January 2005 and 31 December 2010 were included. One insemination was performed per oestrus. Whelping and litter size following CAII were recorded. Potential determinants of these outcomes were assessed. Of 238 inseminations performed, 174 (73.1%) resulted in whelping. The known litter size ranged from 1 to 16 pups (mean ± SD 6.12 ± 3.12 pups). From univariable analyses, progressive motility percentage was the only variable significantly associated with odds of whelping (P = 0.020); bitch parity and weight were associated with litter size when adjusted for each other (P = 0.035 and 0.003, respectively). Inseminations performed with >200 × 10(6) progressively motile sperm were more likely to result in whelping relative to inseminations with >100-200 × 10(6) progressively motile sperm (odds ratio 3.61, 95% confidence interval 0.84-15.5, P = 0.084), and, in a separate model, relative to >75-125 × 10(6) progressively motile sperm (odds ratio 6.09, 95% confidence interval 1.41-26.36, P = 0.016). Whelping rates and litter sizes were similar to other case studies and the experimental reports of CAII. Progressive motility percentage affects the odds of whelping, and litter size is affected by both the weight and parity of the bitch. Importantly, these findings provide some evidence that whelping rates with CAII are not maximised unless more than 200 × 10(6) progressively motile sperm are inseminated.Australian Veterinary Journal 08/2012; 90(8):283-90. · 0.92 Impact Factor
- 02/2012; , ISBN: 978-953-51-0031-7