Preservation and shipment of chilled and cryopreserved dog semen

The transport and artificial insemination of chilled (4 degrees C) and cryopreserved (-196 degrees C) dog semen have gained increasing interest worldwide and have become very popular among dog breeders. Whereas cryopreservation of dog sperm is a complicated and time consuming procedure, which is almost exclusively performed at universities, the chilling of dog semen can be handled by veterinarians in their private practices, provided that the basic knowledge of chilling and diluting semen is acquired. Immediately after sperm collection, the quality of the fresh sample is evaluated and recorded before diluting in an appropriate extender. Subsequently, the diluted semen is gradually chilled to 4 degrees C. It can be stored at 4 degrees C for several days or transported in a thermos flask, a styrofoam box or a Minitube neopore box. Cryopreserved dog sperm is mostly transported in a dry-shipper container. The rules and legislation for the shipment of chilled and frozen dog semen are rather complicated. They differ between almost every country and may change over time. To comply with all the administrative procedures, it is necessary to plan the transport of semen well in advance

Successful cryopreservation of African wild dog (Lycaon pictus) spermatozoa: towards developing the frozen zoo

Sperm freezing and artificial insemination can aid species management and conservation of the African wild dog (Lycaon pictus). Freezing attempts have previously been unsuccessful with sperm motility dropping to nearly 0% within 2 h of thawing. We examined the quality of wild dog spermatozoa subjected to 2 routine canine cryopreservation protocols: 1) 1-step dilution in TRIS-egg yolk extender containing 8% glycerol and 20% egg yolk; and 2) 2-step dilution in TRIS-egg yolk extender to a final concentration of 5% glycerol, 20% egg yolk and 0.5% Equex STM. Protocol 2 showed a significantly higher post-thaw viability, acrosome integrity and longevity of spermatozoa with motility present for up to 8 h after thawing; making it suitable for sperm banking and downstream use in species management by artificial insemination for the first time

Social dominance does not affect semen quality in African wild dogs (Lycaon pictus)

Sperm banking and artificial insemination could benefit conservation of endangered African wild dogs (AWD). However, it is not clear whether their strict dominance hierarchy causes subfertility in subdominant males that typically do not breed. Our study investigated the effect of dominance on male reproductive parameters including: faecal glucocorticoids (fGCMs) and androgens (fAMs), testis and prostate volume, preputial gland size, semen collection success, and the number, motility, morphology, viability, acrosome integrity (PSA-FITC), and DNA integrity (TUNEL) of spermatozoa collected by electroejaculation. Samples were obtained from n=12 captive AWDs (4 US packs) in the pre-breeding season and n=28 captive AWDs (n=11 from 4 US packs; n=17 from 3 Namibian packs) in the breeding season. Male hierarchy was clearly determined by behavioural observations in all but 1 Namibian pack. Data were grouped by dominance status and means compared by ANOVA or t-test. P≤0.05 was significant. In the pre-breeding season, there was no significant difference in body weight, fGCMs, fAMs, or prostate and testis volume between dominance groups. Semen was successfully collected from all alphas but only half the subdominants; with urine contamination negatively associated with dominance. Sperm quality was low (17.3 ± 10.2% total motility, 12.8 ± 8.5% progressive motility, 27.4 ± 11.5 x 106 ejaculated spermatozoa, 40.6 ± 9.8% normal morphology, 63.1 ± 5.1% viability, 72.6 ± 5.2% acrosome integrity) with no difference observed in any parameter except progressive motility and normal sperm morphology; which were significantly lower in subdominants (27.7 ± 16.8% vs. 0.0 ± 0.0% and 59.8 ± 13.0% vs. 21.4 ± 5.7%). From pre-breeding to breeding season, testis and prostate volume increased significantly; particularly in beta and gamma males respectively. Prostate volume was higher in alpha than beta males (16.0 ± 6.4 cm3 vs. 5.7 ± 1.4 cm3), but testis volume, body weight, fAMs and fGCMs did not differ between dominance groups (12.0 ± 0.9 cm3, 28.5 ± 0.8 kg, 0.51 ± 0.07 µg/g dry weight - DW, 30.6 ± 2.3 ng/g DW). Semen was successfully collected from 75% of males; with reduced urine contamination. Collection success, urine contamination and preputial gland size were not associated with dominance. Sperm quality improved with significantly greater number, viability, and total motility. However, sperm quality did not differ between dominance groups (47.4 ± 6.7% total motility, 30.5 ± 5.8% progressive motility, 32.3 ± 9.2 x 106 ejaculated spermatozoa, 50.9 ± 5.2% normal morphology, 74.4 ± 4.2% viability, 85.6 ± 3.0% acrosome integrity, 99.7 ± 0.1% DNA integrity). In conclusion, subdominant males are at higher risk of urine contamination and have lower sperm motility and normal morphology when semen is collected in the pre-breeding season. However, their semen is of similar quality to dominant males in the breeding season, indicating that reproductive suppression of subdominant males is only behavioural. Thus, AWD males of all social ranks in the breeding season are suitable candidates for sperm banking