Sex and PRNP genotype determination in preimplantation caprine embryos.

The objective of this study was to test the accuracy of genotype diagnosis after whole amplification of DNA extracted from biopsies obtained by trimming goat embryos and to evaluate the viability of biopsied embryos after vitrification/warming and transfer. Whole genome amplification (WGA) was performed using Multiple Displacement Amplification (MDA). Sex and prion protein (PRNP) genotypes were determined. Sex diagnosis was carried out by PCR amplification of ZFX/ZFY and Y chromosome-specific sequences. Prion protein genotype determination was performed on codons 142, 154, 211, 222 and 240. Embryos were collected at day 7 after oestrus and biopsied either immediately after collection (blastocysts and expanded blastocysts) or after 24 h of in vitro culture (compacted morulae). Biopsied embryos were frozen by vitrification. Vitrified whole embryos were kept as control. DNA of biopsies was extracted and amplified using MDA. Sex diagnosis was efficient for 97.4% of biopsies and PRNP genotyping was determined in 78.7% of biopsies. After embryo transfer, no significant difference was observed in kidding rate between biopsied and vitrified control embryos, whereas embryo survival rate was different between biopsied and whole vitrified embryos (p = 0.032). At birth, 100% of diagnosed sex and 98.2% of predetermined codons were correct. Offspring PRNP profiles were in agreement with parental genotype. Whole genome amplification with MDA kit coupled with sex diagnosis and PRNP genotype predetermination are very accurate techniques to genotype goat embryos before transfer. These novel results allow us to plan selection of scrapie-resistant genotypes and kid sex before transfer of cryopreserved embryo.

Efficacy of two types of vaginal sponges to control onset of oestrus, time of preovulatory LH peak and kidding rate in goats inseminated with variable numbers of spermatozoa.

In small ruminants, progestagen-impregnated vaginal devices (sponges) are useful tools to manage reproduction irrespective of season and to the application of timed artificial insemination (AI). A novel progestagen releasing vaginal-controlled release device (Chronogest CR), loaded with less (20mg) cronolone using proprietary procedures, was developed and its efficacy (synchronising ability, fertility and prolificacy following sponge removal) evaluated versus the existing Chronogest sponge containing 45 mg of cronolone in goats. Females (n=199) were maintained in field conditions and inseminated with graded amounts of spermatozoa at two stages of the year (breeding and non-breeding seasons). The use of the new Chronogest CR sponge was associated with an earlier initiation of the LH surge (28.7h versus 30.8h following sponge removal, P<0.01). A similar degree of synchronisation of the LH surge was obtained with both types of sponges. In both treatment groups, a longer time interval between sponge removal and the LH surge was noted in females with high milk production. Fertility and prolificacy were high and unaffected by the type of sponge used or the amount of spermatozoa inseminated. It is concluded that the new Chronogest CR sponge allows a reduction of the progestagen load from 45 to 20mg without detrimental effects on synchronisation, fertility and prolificacy.

Four years of induction/synchronization of estrus in dairy goats: effect on the evolution of eCG binding rate in relation with the parameters of reproduction.

Ninety-eight Alpine goats of two herds were followed over 4 years in a program of annual artificial insemination after estrus induction/synchronization, including progestagen administration (vaginal sponge) followed by prostaglandin analog and equine chorionic gonadotrophin (eCG) 48 h before sponge removal. Goats were sampled every 4 hours from the 16th to the 56th following sponge removal, for determination of LH surge and tested for estrus by the presence of a buck. Seven days after AI, endoscopic examination of the ovaries was performed to determine the number of corpus lutea. Pregnancy diagnosis was performed at day 21-22 post AI by determination of plasma progesterone and at day 40-45 by ultrasonography. Parturition, number and sex of kids were recorded. All the goats were sampled before and after each treatment, for anti-eCG antibodies screening. Statistical analysis of the results clearly established a significant effect of the treatments on anti-eCG antibodies. Time of estrus and LH surge were significantly different between herd. The antibodies significantly delayed the time of coming out of estrus as well as the time of LH surge. Two antagonistic effects were evidenced: first, the delayed of time of estrus and time of LH surge in relation with the immune reaction to eCG; secondly, the ahead of time of estrus and time of LH surge during the years of treatment, identical to both herd. The antibodies negatively influenced the percentage of ovulating females as well as kidding rate. Finally, no effect of antibodies on prolificacy was found.