B. Leboeuf

Production and storage of goat semen for artificial insemination.

Environmental influences on reproduction and semen production in the buck, the problem of interaction between seminal plasma and egg yolk or milk constituents in diluent, liquid storage and processing of semen for freezing are discussed. A review is given on the use of frozen-thawed semen for artificial insemination (AI) in spontaneous and induced oestrus and factors influencing the fertility.

Synchronization of estrus in goats: the relationship between time of occurrence of estrus and fertility following artificial insemination.

The fertility rate for goats following artificial insemination (AI) is usually analyzed according to herd or treatment groups. However, these general information are insufficient to allow identification of specific factors which affect this individual reproductive performance. In the present experiment 640 dairy goats were used to analyze to what extent the interval from sponge removal to estrus affects the results of AI, performed at a predetermined time following sponge removal. Estrus occurred in 98.1% of experimental animals between 24 and 72 hours after sponge removal. The fertility rate was lower for goats that came into estrus later than 30 hours after sponge removal (33.3%, n = 108 than for goats that exhibited estrus earlier (65.0%, n = 520; P<0.001). The occurrence of late estrus is not age dependent, but it increases with the number of treatments that an individual animal has previously received. These results show that the low fertility rate observed in some herds after synchronization of estrus and AI may be related to the high proportion of goats with a late occurrence of estrus, and this phenomenon increases in animals that are treated repeatedly.

Artificial insemination of dairy goats in France

Abstract In France, goat Artificial Insemination (AI) plays an important role in the milk intensive production system to control reproduction and, in conjunction with progeny testing, to improve milk production. About 60 000 goats were inseminated in 1996, 95% of them before the breeding season with deep frozen semen after induction of oestrus and ovulation by hormonal treatment. The efficiency of this method was about 65% of kidding rate. In 1996, 290 000 lactating goats were officially recorded. The selection scheme, aimed at improving protein yield and protein content, benefits from the sampling program for young AI buck. AI allows higher selection intensities for male selection, adequate genetic evaluations through genetically connected herds, and monitoring and selection of bucks for major genes like the αs1-casein gene. In order to further develop AI, a cooperative action between the partners of the dairy goat industry (Research, Agricultural extension and Farmer organizations) has operated since 1992 with the aim of reducing the variability of fertility rates. This program includes improvement of the control of reproduction by hormonal treatments, AI conditions, embryonic mortality and pseudopregnancy.

Synchronization of estrus in goats: The relationship between eCG binding in plasma, time of occurrence of estrus and fertility following artificial insemination

Abstract Radioimmunoassay (RIA) was used to measure plasma eCG binding in dairy goats (n = 524) at the beginning of a progestagen/eCG treatment and 25 d after eCG administration. The eCG binding was not dependent on the age of the females but increased with the number of treatments they had previously received (3.4 % ± 4.8, n = 47 vs 9.6 % ± 13.2, n = 249; mean ± SD; P

Successful direct transfer of vitrified sheep embryos

The use of a simple cryopreservation method, adapted to direct transfer of thawed embryos may help to reduce the costs of embryo transfer in sheep and increase the use of this technique genetic improvement of this species. Two experiments were made to test a vitrification method that is easy to apply in field conditions. All embryos were collected at Day 7 of the estrous cycle of FSH-stimulated donor ewes and were assessed morphologically, washed in modified PBS and incubated for 5 min in 10% glycerol, for 5 min in 10% glycerol and 20% ethylene glycol and were transferred into the vitrification solution (25% glycerol and 25% ethylene glycol). All solutions were based on mPBS. Embryos were loaded in straws (1 cm central part, the remaining parts being filled with 0.8 M galactose in mPBS) and plunged into liquid N2 within 30 sec of contact with the vitrification solution. The straws were thawed (10 sec at 20 degrees C) and the embryos were either transferred directly or after 5 min of incubation in the content of the straw (followed by washing in PBS) into the uterus of a recipient ewe. In Trial 1, the pregnancy rates at term (72 vs. 72%) as well as the embryo survival rates (60 vs 50% respectively) were not different between fresh (n = 48 embryos) and vitrified (n = 50) embryos. In a second trial no difference was observed between vitrified embryos transferred after in vitro removal of the cryoprotectant (n = 86 embryos) or directly after thawing (n = 72) both in terms of lambing rate (67 vs. 75%, respectively) and embryo survival rate (lambs born/embryos transferred; 49 vs. 53%). This method of sheep embryo cryopreservation provided high pregnancy and embryo survival, even after direct transfer of the embryos.

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.

Effect of native phosphocaseinate on the in vitro preservation of fresh semen

The fertilization capacity of goat sperm stored in milk extenders is approximately 12-24h. Long-term storage of goat sperm (up to 3 days) is desirable as it would confer greater flexibility to breeding farms. The aim of this study was to evaluate in vitro motility parameters of buck spermatozoa for up to 7 days of storage using skim milk or chemically defined extender supplemented with native phosphocaseinate (NPPC). Four experiments were conducted to determine optimum temperature (4 or 15 degrees C) and storage conditions (aerobic versus anaerobic), the effect of seminal plasma on sperm survival, the optimal concentration of NPPC and the effect of beta lactoglobulin (BL). Both skim milk and NPPC were found to be more efficient for preserving goat sperm at 4 degrees C than at 15 degrees C (P<0.01). Furthermore, when sperm was stored at 4 degrees C, no detrimental effects of seminal plasma were observed. Our results showed that motility parameters can be maintained with success until Day 4. However, NPPC-based extenders extend the in vitro survival to 7 days of storage. The optimal concentration of NPPC for the preservation of sperm cells for 4 days of storage was 81g/l and for 7 days of storage was 81 and 54g/l. No effect of the supplementation of the NPPC extender with BL was found.

A new method for controlling the precise time of occurrence of the preovulatory gonadotropin surge in superovulated goats

In goats treated to induce superovulation, insemination at a predetermined time after the end of progestagen treatment leads to a low fertilization rate. To solve this problem we developed a new treatment based on the control of the occurrence of the endogenous LH peak with a GnRH antagonist (Antarelix). The first experiment was designed to determine the dose of LH required to mimic a spontaneous LH preovulatory discharge; the injection of 3 mg, i.v. of pLH induced a peak of the same amplitude and duration as the spontaneous peak. Subsequently, in the second experiment, we compared 2 doses of Antarelix (0.5 and 1 mg, sc) administered 12 h after sponge removal (9 goats/treatment group). The dose of 0.5 mg was selected for further experiments because it was effective in the inhibition of the endogenous LH peak and had no detrimental effect on the quality of embryos. In the final experiment, 48 goats received the new treatment and were inseminated (intrauterine) only once 16 h after LH injection; 41 were flushed and produced 5.3 +/- 4.5 (m +/- SD) transferable embryos. The developmental stage and the number of cells/embryo were within the range that has been reported for embryos produced with conventional treatments. In conclusion, with the described method, it is possible to inseminate goats at a predetermined time without decreasing the number of transferable embryos. This technique will encourage the development of embryo transfer within genetic programs, and it will be a valuable tool for the production of zygotes for gene transfer.

Genetic and non-genetic parameters of several characteristics of production and semen quality in young bucks

The objective of this study was to evaluate genetic and non-genetic factors influencing characteristics of young buck semen production using a multivariate model that takes into account the longitudinal structure of data. Data were collected from 1989 to 2002 at two French A.I. centres. The data corresponded to 13151 and 9206 ejaculates of 758 Alpine and 535 Saanen bucks respectively, collected at the beginning of the first breeding season (September-December). The semen volume, the total number of spermatozoa, the concentration, the motility score of spermatozoa after freezing and the percentage of motile spermatozoa after freezing were registered for each ejaculate. Within-breed heritabilities and repeatabilities were estimated using a multivariate animal model using a power spatial covariance structure for environmental effect. For all characteristics and the two breeds, the main source of variation was the year-month interaction that interacted with the centre. We observed a decrease in years of motility score after freezing. Age and frequency of collection had a significant effect on semen volume and number of spermatozoa for both breeds, and on concentration of spermatozoa for the Alpine breed. No effect of these factors was found on the characteristics observed after freezing. Heritabilities for concentration, number of spermatozoa, semen volume, motility score after freezing and percentage of motile spermatozoa after freezing per ejaculate were respectively, 0.32, 0.15, 0.25, 0.12 and 0.05 for the Saanen breed and 0.34, 0.25, 0.29, 0.17 and 0.03 for the Alpine breed. Genetic correlations between volume and number of spermatozoa were respectively, 0.74 for the Alpine breed and 0.86 for the Saanen breed. Further study is required to compare the semen characteristics of young bucks with their mature production.

The value of the percentage of motile sperm in predicting a significant portion of the fertility variation of frozen-thawed buck semen

Prediction of the future fertility of a given ejaculate with a simple laboratory test is still considered a real issue in domestic mammal breeding. This study showed that a subjective assessment of the percentage of motile spermatozoa, measured 120 min after thawing (mob120), can predict a significant part (∼50%) of the variation of the future fertility of buck ejaculates. The predictive model was calculated using a calibration data set composed of 40 ejaculates from four Alpine and six Saanen bucks. A fertility trial using split ejaculates was conducted in order to estimate ejaculate fertility. Taken into account were the herd within breed factor and the year, month, and inseminator factors. On average, one ejaculate was used to inseminate two females per herd in 10 different herds. This calibration set allowed us to choose the mob120 variable among a set of laboratory tests: mitochondrial activity, acrosomal status, membrane integrity, osmotic resistance test assessed by flow cytometry, velocity and motion characteristics assessed by computer-assisted sperm analysis, visually assessed percentage of motile, and motility score measured 5 and 120 min after thawing. For the calibration step, the best model used the logarithm of mob120 and gave a correlation coefficient of 0.71 between the field fertility and the predicted fertility and a standard error of 0.17. We tested this model on 3 different validation data sets adding up to 95 ejaculates that were all different from those of the calibration data set. The correlation coefficients between field fertility and predicted fertility were always significant and the bias corrected standard error ranged from 0.15 to 0.18 on these validation data sets. A Monte Carlo simulation showed that about 20% of the fertility variation remained to be explained.