Fertility results of artificial inseminations performed with liquid boar semen stored in X-cell vs BTS extender.

The objective of the present field study was to compare the fertility results for boar semen diluted in X-cell stored up to 4-5 days before artificial insemination (AI) with semen diluted in Beltsville thawing solution (BTS) used for AI following 2-3 days of storage (where the first day being the collection day). A total number of 2601 double inseminations in Norwegian herds were included in this two-trial study. All the boars used in the study were mature cross-bred Norwegian Landrace x Duroc (LD), which were routinely used for AI in Norway. The inseminated gilts and sows were Norwegian Landrace x Yorkshire (LY). The AI doses contained 2.5 billion spermatozoa, and consisted of a mixture of semen from three, occasionally four, boars (i.e. heterospermic semen). Fertility was measured in terms of the likelihood of farrowing and subsequent litter size. The fertility of the semen in both of the extenders was satisfactory and no significant differences were found either in semen stored 4-5 days in X-cell compared with 2-3 days in BTS or in semen stored 2-3 days in X-cell compared with 2-3 days in BTS. The storage capability findings for the long-term extender X-cell could significantly simplify the practical issues of semen production and the distribution of AI doses containing 2.5 billion spermatozoa. However, in pig production systems where all semen is used within 2-3 days, the short-term extender BTS is as good as the more expensive extender X-cell.

Within and between breed differences in freezing tolerance and plasma membrane fatty acid composition of boar sperm.

The response of sperm to cryopreservation and the fertility of frozen-thawed semen varies between species. Besides species differences in sperm physiology, structure and biochemistry, factors such as sperm transport and female reproductive tract anatomy will affect fertility of frozen-thawed semen. Therefore, studying differences in sperm cryotolerance between breeds and individuals instead of between species may reveal sources of variability in sperm cryotolerance. In the present study, the effect of cooling, re-warming and freezing and thawing on plasma membrane and acrosome integrity of sperm within and between Norwegian Landrace and Duroc breeds was studied. Furthermore, the relation between post-thaw survival rate and fatty acid composition of the sperm plasma membranes was investigated. Flow cytometry assessments of plasma membrane and acrosome integrity revealed no significant differences between breeds; however there were significant male-to-male variations within breeds in post-thaw percentages of live sperm (plasma membrane intact). The most abundant fatty acids in the plasma membranes from both breeds were palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1, n-9), docosapentaenoic acid (22:5, n-6) and docosahexaenoic acid (22:6, n-3). The ratio of sigma operator 22:5, n-6 and 22:6, n-3/ sigma operator all other membrane fatty acids was significantly related to survival rate (plasma membrane integrity) of sperm for both Norwegian Landrace (correlation coefficient (r(s)) = 0.64, P < 0.05) and Duroc (r(s) = 0.67, P < 0.05) boars. In conclusion, male-to-male differences in sperm survival rate after freezing and thawing may be partly related to the amount of long-chain polyunsaturated fatty acids in the sperm plasma membranes.

A retrospective study on effects of storage time of liquid boar semen on reproductive performance in Norwegian swine.

The objective of this retrospective field study was to determine the effects of storing, for up to 62 h, heterospermic and homospermic semen in the short-term extender Beltsville thawing solution (BTS), on reproductive performance in Norwegian swine of four different breed combinations. The study was based on fertility records after single or double inseminations with semen collected at an AI station in Norway from January 1998 to June 2001. Increasing the duration of storage of homospermic semen, but not heterospermic semen, from an interval of 4-14 h to an interval of 52-62 h, was associated with a 0.5 piglet reduction in litter size. There were differences in reproductive performance among breed combinations that appeared to be associated with duration of semen storage. In conclusion, prolonged semen storage decreased reproductive performance; the extent varied among breeds and was prevented by the use of heterospermic versus homospermic semen.

Effects of in vitro storage time and semen-extender on membrane quality of boar sperm assessed by flow cytometry.

The Norwegian AI company Norsvin has used the short-term semen-extender BTS to extend and store boar semen since the late 1980s. Fertility results have been consistent when extended semen has been used for AI within 3 days after collection, however, from a production and economic point of view it is preferable that semen stored for up to 5 days can be used. The aim of this study was to compare membrane quality of sperm stored in BTS for 3 days with sperm stored in the long-term semen-extenders Androstar, Mulberry III and X-cell for 5 days. Using a split-sample design, plasma membrane- and acrosome-integrity were assessed flow cytometrically by use of Yo-Pro-1 and PNA-FITC, and fluidity and phospholipid asymmetry of the membrane were assessed by use of MC540 and Annexin V-FITC. Due to observed sperm fragmentation in Androstar after Day 1, the data for Androstar were excluded from the analyses. After 5 days of storage, the membrane quality of X-cell-stored sperm was not statistically different from that of sperm stored in BTS for 3 days, while membrane quality of sperm stored in Mulberry III was statistically better on Day 5 compared to BTS on Day 3. In conclusion, Mulberry III and X-cell preserve sperm quality, as well as that of BTS on Day 3, for up to 5 days after collection.

Reproductive performance of gilts and sows in temperate versus subarctic and arctic zones of Norway.

The reproductive performance of gilts and sows from two regions in Norway was investigated in a retrospective analysis of data from the litter recording system. In the Northern region (North; between 65 degrees N and 71 degrees N), there are extreme shifts in natural photoperiod between winter and summer. In the Southern region (South; between 59 degrees N and 60 degrees 30'N), photoperiodic changes are less dramatic. Gilts were 8 days older at first mating or insemination in the North than in the South (P<0.01). A significantly lower proportion of sows in the North were mated or inseminated within 5 days post-weaning than in the South, a difference present both among primiparous and multiparous sows (P<0.01). Overall farrowing rate in the North was lower than in the South, but litter size (total number born) among those pigs that farrowed was larger. After correction for year, month, breed and age at first service, there were still lower odds of farrowing for gilts in North than in South. Neither for primiparous nor multiparous sows were regional differences in farrowing probability significant when year, month, breed and weaning to service interval were included in the model. Gilts and primiparous sows had a lower probability of farrowing following insemination during summer or autumn months, but service month was not significantly related to the farrowing probability of multiparous sows. For gilts, litter size was positively related to age at first service. For sows, litter size was lowest at weaning to service intervals between 6 and 10 days. Total numbers of piglets born per litter were estimated to be 0.36, 0.38 and 0.55 larger in the North than in the South (differences in least square means; gilts, primiparous sows and multiparous sows, respectively) (P<0.01). Litter size was lower after service during natural long photoperiod than during the rest of the year.

The importance of farrowing to service interval in sows served during lactation or after shorter lactation than 28 days.

In a retrospective study, based on data from the national litter recording system, farrowing rate and litter size of sows served (inseminated or mated) during the lactation period (n = 574) or after a lactation period shorter than 28 days (n = 14,219) were analysed. The results were compared with the corresponding figures for sows with lactation length between 28 and 35 days and weaning to first service interval of 4 or 5 days (reference group; n = 41,741). The farrowing rate of the reference group was 80.9% and subsequent litter size was 13.7 total piglets born. Among sows served prior to weaning, the farrowing rates and litter sizes were significantly lower for those served earlier than 22 days post-farrowing compared to those served later (P < 0.05). Shorter lactations than 28 days and service within 10 days post-weaning led to lower farrowing rates than in the reference group (P < 0.01). Significant differences were seen after different lactation lengths. After correction for weaning to service interval, preceding litter size weaned, parity, breed and the interaction between parity and breed, litter size was significantly and positively associated with the preceding lactation length. The study shows that service within the first 3 weeks post-farrowing results in reduced reproductive performance.

Oestrone sulphate measurements for the prediction of small or large litters in pigs.

Serum from 88 pregnant sows and gilts was sampled 24 and 28 days after their first insemination or mating day. The oestrone sulphate (E1S) concentration in the samples was assessed with a commercially available radioimmunoassay kit modified for use with swine serum. The first aim was to test whether it was possible to predict litters of total number <10 piglets at term. The second aim was to compare the use of day 24 or day 28 samples, or of both, in this prediction. Day 24 E1S levels were positively correlated with litter size at term (R2 = 0.26; p <0.001). E1S levels on day 28 were correlated with the levels on day 24 in the same animals but could not be used for prediction of large or small litters. The odds ratio for a small litter size was 0.16 (p <0.01). This means that odds for a litter size <10 piglets decreased by 84% when E1S levels increased by 1.0 ng/ml.

A preliminary study on the effect of dietary supplementation with cod liver oil on the polyunsaturated fatty acid composition of boar semen.

Eight mature Norwegian Landrace boars, of proven fertility and in routine semen production for AI, were fed individually with the same basic diet for 9 weeks. One group of 4 animals served as the control, the remaining 4 boars received a daily supplement of 75 ml cod liver oil (CLO-group). Fifteen consecutive semen samples were collected from each boar. The fatty acid composition of the semen was determined, and the content of the 15 most numerous fatty acids with a chain length longer than 12 carbon atoms was followed over time. In both groups, the proportion of 16:1n-7 decreased significantly, while 16:0 and 22:6n-3 (DHA) increased. By the end of the experiment, DHA had tended to increase and 22:5n-6 to decrease to a greater extent in the CLO-group. A significant difference between the groups was seen for one n-6 PUFA (22:4n-6), which remained unchanged in the control group but decreased in the CLO-group. No change was seen in docosapentaenoic acid (22:5n-3) and eicosapentaenoic acid (20:5n-3) was not found in any sample. These results indicate that CLO supplementation affects the fatty acid composition of boar semen. There were no significant differences in the non-return rates (4-25 days) between the two groups before, during or after the experiment.

In vivo fertilizing capacity of deep frozen boar semen packaged in plastic bags and maxi-straws.

Pooled ejaculates from six fertile boars were frozen under controlled conditions in Teflon FEP-film plastic bags (5 ml) and maxi-straws (2.5 ml) using 3% glycerol as cryoprotectant. The percentages of both post-thaw motility and normal apical ridges were significantly higher (P less than 0.001) for the bags (54.5 and 75%) than for the maxi-straws (40.1 and 59.4%) respectively. For evaluation of the in vivo fertilizing capacity of the frozen-thawed spermatozoa, 26 gilts were inseminated once 24 h after the first observation of standing reflex in their second oestrus, with 5 ml of semen (containing 5 billion spermatozoa) reconstituted in 80 ml of BTS from either bags or maxi-straws. Ova were recovered from the oviducts/uteri 2-4 days following insemination and examined for cleavage and sperm binding to the zona pellucida (ZP). Significantly higher rates (P less than 0.02) of fertilized ova were found in the bag-inseminated (75%) than in maxi-straw inseminated gilts (63%); and similarly their ova had significantly more spermatozoa in the ZP, irrespective of whether they were fertilized or nonfertilized. This study confirmed that the plastic bags are suitable and may be used for packaging single insemination doses of deep frozen boar semen for routine A.I. work.

Effect of melatonin implants on reproduction in the male silver fox (Vulpes vulpes).

In June 1987, when the testes were fully regressed, 5 males were given s.c. implants of 40 mg melatonin. The same treatment was repeated in August and October 1987. Five males served as controls. Plasma concentrations of melatonin increased significantly in treated males and were still elevated at the end of the study in April 1988. The changes in testicular volume and blood plasma concentrations of testosterone in response to GnRH indicated that melatonin administration promoted testicular development. However, testicular regression was observed earlier in treated than control animals, perhaps because of refractoriness to melatonin or to a down-regulation of melatonin receptors. Semen was collected and frozen in November 1987, 2 months ahead of the natural breeding season, from the melatonin-treated males, and 10 blue fox vixens were inseminated the following breeding season: 9 vixens conceived, and the average litter size was 7.6 +/- 0.5. The results demonstrate that melatonin treatment initiated during exposure to naturally long days (a) promotes testicular development in a way similar to an artificial short photoperiod and (b) may induce a refractory condition after an extended period of treatment.

Photoperiodic regulation of reproduction in the male silver fox (Vulpes vulpes).

Six silver fox males were exposed to short days (6L:18D) from February, when the testes were fully developed, until June 1986 (Group 6L). Eight males maintained in natural daylight served as controls (Group N). Histological sections from the testes of 2 males in Group 6L killed in June indicated full spermatogenic activity. Three blue fox vixens inseminated the following year with semen collected and frozen in June from 3 males in Group 6L failed to produce litters. One possible explanation for the reproductive failures could have been that the high environmental temperatures in June influenced semen quality. There was no significant difference (P greater than 0.05) in LH release in response to GnRH stimulation in June, but testosterone response to LH release was significantly higher (P less than 0.01) in animals subjected to a restricted photoperiod, demonstrating that testicular testosterone production was maintained longer than in control animals. Two males in Group 6L were retained in 6L:18D from June until December 1986 and then exposed to natural daylight until the end of the study in May 1987 (Group 6L:6L:N). These males started to shed their winter coat and showed clinical signs of testicular regression in December, i.e. after approximately 11 months exposure to 6L:18D. The 2 remaining males in Group 6L were moved to cages with natural daylight in June 1986, where they were kept until the end of the experiment (Group 6L: N:N). These males displayed testicular regression soon after the change in photoperiod but maintained their capacity for testicular redevelopment during the following breeding season.(ABSTRACT TRUNCATED AT 250 WORDS)

Electron microscopical studies of membrane injuries in blue fox spermatozoa subjected to the process of freezing and thawing.

Disintegration of blue fox sperm membranes is studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In unfrozen spermatozoa studied by SEM, the plasmalemma and the acrosome appeared to be intact, except for a few cases of disruption of the former structure at the anterior part of the head. In semen frozen in 0.5-ml plastic straws by use of N2 vapor after dilution with Tris-fructose-citric acid with 8 vol % glycerol and 20 vol % egg yolk and thawed at 70 degrees C for 8 sec, the spermatozoa displayed different degrees of membrane damage. These alterations could be classified into three main categories of which the first included only minor changes in the plasmalemma, but vesiculation and disintegration of the outer part of the acrosomal membrane. In the second category (also the most frequent one) the outer part of the acrosomal membrane was extensively vesiculated, and the plasmalemma was discharged proximal to the equatorial segment. Extensive loss of plasmalemma and complete absence of the outer part of the acrosomal membrane characterized the last category of membrane damage. The functional implications of the three categories of membrane alterations are discussed.

Preliminary studies of the effects of bromocriptine on testicular regression and the spring moult in a seasonal breeder, the male blue fox (Alopex lagopus).

Bromocriptine administration in the form of slow-release injections to male blue foxes during March-May abolished the normal spring rise in plasma prolactin concentrations seen in May and June. The spring moult was prevented and the treated animals retained a winter coat of varied quality and maturity until the end of the study in August. Plasma testosterone concentrations fell normally from March until August. Testicular regression was, however, delayed, although there were individual variations in response. Estimation by DNA flow cytometry in early July of the relative numbers of haploid, diploid and tetraploid cells in the testis showed that, in the treated animals, 74-80% of the cells were haploid (maturing germinal cells), 4-6% tetraploid (mainly primary spermatocytes) and the rest diploid cells (somatic cells and the remaining germinal cell types). In the control males, however, no haploid cells were detected and the majority of cells were diploid (93-99%). At castration in August, histological examination revealed various stages of testicular regression in the treated and control animals.