Boar semen cryopreservation: State of the art, and international trade vision.

Biosecurity is a major concern in the global pig production. The separation in time of semen collection, processing and insemination in the pig farm is a few days for chilled semen but it can be indefinite when using cryopreserved semen. Field fertility results of boar cryopreserved semen are close to chilled semen, which makes it a valuable resource for the establishment of semen genebanks, long-distance semen trade, and the implementation of other technologies such as the sex-sorted semen. But cryopreserved semen is far from being routine in pig farms. The most recent research efforts to facilitate its implementation include the use of additives before freezing, or in the thawing extender. Long-term preserved semen trade is a biosecurity challenge. To harmonize international trade of germplasm, the World Organization of Animal Health (WOAH) established a regulatory framework for all member countries. The present paper aims to review the latest advances of boar semen cryopreservation with special focus on the benefits of its inclusion as a routine tool in the pig industry. We also review recently reported field fertility results of cryopreserved semen, its international trade compared to chilled semen, and the regulatory framework involved. Boar cryopreserved semen is a valuable tool to control biosecurity risk, implement other technologies, and facilitate international trade. Research already demonstrated good field fertility results, but it still represents less than 0.1 % of the international trade. As boar cryopreserved semen gets closer to implementation, the correspondent authorities are reviewing the trade rules.

Low density Porcicoll separates spermatozoa from bacteria and retains sperm quality.

Antibiotics are added to semen extenders to control the growth of bacteria contaminating semen during collection but may contribute to the development of antibiotic resistance. An alternative would be physical separation of spermatozoa from bacteria. The objective of the present study was to evaluate two low densities of Porcicoll for removal of bacteria, and for their effect on sperm recovery and sperm quality. Semen was collected from boars at a commercial station. Aliquots of 8 extended ejaculates were subjected to colloid centrifugation through 20% Porcicoll (P20) and 30% Porcicoll (P30) in 500 mL tubes and then stored at 17 °C. Microbiological examination and sperm quality evaluation (computer assisted sperm analysis and flow cytometry) were carried out on controls and all colloid-selected samples immediately after preparation and again after storage for 3 and 7 days. The microorganisms found were mainly bacteria from the environment, gut or skin. There was a considerable reduction or complete removal of some bacteria by both colloids. Recovery rates were 86% for P20 and 81% for P30. Sperm quality was not adversely affected by colloid centrifugation on day 0, and thereafter showed a more gradual deterioration in colloid centrifuged samples than in controls, possibly due to lower bacterial contamination. There were no differences in sperm quality between the two colloid treatments. Thus, these results show that contaminating bacteria in semen can be controlled by centrifugation through low density colloids.

Altrenogest treatment before weaning improves litter size in sows.

The study aimed to assess whether altrenogest treatment, fed before weaning (from -8 to -2 days), could improve fertility of sows showing reproductive seasonality. Ninety sows (50 in winter-spring [WS] and 40 in summer-autumn [SA]) were randomly selected and assigned to control (C; 27 in WS and 20 in SA) or altrenogest treatment (A; 23 in WS and 20 in SA) groups. The diameter and number of ovarian follicles were transrectally scanned at the onset of oestrus. Oestrus was evaluated twice daily from weaning to day 8 post-weaning. Sows in oestrus were post-cervically inseminated at 0 and 24 hr after the onset of oestrus with liquid stored semen (1.5 × 109 sperm/doses), and farrowing rates (FR) and total piglets born (LS) were recorded. More (p < .01) sows showed no signs of oestrus within 8 days after weaning in SA (30%) than in WS (2%), without differences between A and C groups. The diameter (cm) of the follicles at the onset of oestrus was larger in A than in C sows (0.76 ± 0.01 vs 0.73 ± 0.01; p < .01), irrespective of the season. No differences in the number of follicles were found. FR did not differ between seasons and groups, being always above 85%. LS was larger (p < .01) in A (14.00 ± 0.46) than C (12.27 ± 0.44) sows, irrespective of the season. In conclusion, a short-term altrenogest treatment at the end of lactation improves the total number of piglets born from weaned sows, probably by promoting a better and more homogeneous follicular development at the start of oestrus.

Will AI in pigs become more efficient?

AI is commercially applied worldwide to breed pigs, yielding fertility outcomes similar to those of natural mating. However, it is not fully efficient, as only liquid-stored semen is used, with a single boar inseminating about 2000 sows yearly. The use of liquid semen, moreover, constrains international trade and slows genetic improvement. Research efforts, reviewed hereby, are underway to reverse this inefficient scenario. Special attention is paid to studies intended to decrease the number of sperm used per pregnant sow, facilitating the practical use of sexed frozen-thawed semen in swine commercial insemination programs.

Boar Differences In Artificial Insemination Outcomes: Can They Be Minimized?

In Western countries, where pig breeding and production are intensive, there is a documented variability in fertility between farms with boar-related parameters only accounting to 6% of this total variation of in vivo fertility. Such low boar effect could be a result of the rigorous control of sires and ejaculates yielding AI-doses exerted by the highly specialized AI-centres that monopolize the market. However, some subfertile boars pass through these rigorous controls and consequently reach the AI-programmes. Here, we discuss why testing young boars for chromosomal defects, sperm nuclear chromatin integrity and in vitro fertilizing ability can be discriminative and economically sound for removing these less fertile boars. Alongside, we discuss why boars differ in the ability of their sperm to tolerate cryopreservation or sex sorting.

Use of frozen-thawed semen aggravates the summer-autumn infertility of artificially inseminated weaned sows in the Mediterranean region.

Improvement of farrowing rate (FR) and litter size (LS) of sows that are AI with frozen-thawed (FT) semen can hardly be reached without identification of the factors behind the high variability seen among trials. Three experiments using weaned (4-d wean-to-estrus interval) multiparous (parity 2 to 7) sows were conducted to evaluate the effect of period of the year on FR and LS of FT-inseminated sows in southern Spain. Sows were grouped into 2 periods of the year: winter-spring (November to April; WS) and summer-autumn (May to October; SA). Ovarian status was monitored by transrectal ultrasonography to record how long before or after ovulation AI was performed (pre-, peri-, or postovulatory AI) and to determine the onset of estrus-to-ovulation interval (EOI). Inseminations were performed using deep intrauterine AI with 1.5 x 10(9) FT sperm per dose. The first experiment was designed to determine the influence of the period of the year on FR and LS of FT semen. Sows (116 in WS and 100 in SA) were AI at 33 and 39 h after the onset of estrus. The period of the year influenced the FR and LS (P < 0.01). Farrowing rate and LS were least in SA (P < 0.05). This pattern of annual variation was similar to that shown by sows on the same farm currently undergoing AI with liquid semen (cervical AI at 12 and 36 h after the onset of estrus with 3 x 10(9) sperm per dose). However, the FR reduction in SA respect to WS was more substantial in sows artificially inseminated with FT (77.6 vs. 50%, P < 0.001) than those artificially inseminated with liquid semen (83.9 vs. 71.8%, P < 0.05). More pre- and less periovulatory AI were performed in SA sows than in WS sows (P 48 h) in SA than in WS (P 0.05). In the Spanish Mediterranean area, a longer EOI during SA negatively influenced the FR and LS of weaned sows after AI. This effect was particularly evident when FT semen was used. These findings were not ameliorated by hormonal induction of ovulation.

Strategies to improve the fertility of frozen-thawed boar semen for artificial insemination.

Although cryopreservation of boar semen for artificial insemination (AI) was developed 35 years ago, cryopreservation conditions and AI strategies are still considered sub-optimal. AI with excessive numbers of frozen-thawed sperm (5-6 x 10(9) cells), still does not achieve fertility levels similar to AI using liquid semen because of reduced sperm survival. Frozen-thawed (FT) spermatozoa have therefore not been the preferred option for commercial breeding programmes. However, substantial progress has been made regarding boar sperm cryopreservation. Adjustment of cooling and re-warming rates to biophysical properties of boar spermatozoa, new sperm package systems and the achievement of accurately consistent freezing of large numbers of samples using programmable freezers have contributed to post-thaw survival rates above 50%, a threshold similar to that used for bull AI-semen. Moreover, these post-thaw sperm survival rates are consistent within a large population of boars selected for sperm freezability potential, as occurs with AI-bull sires. When such post-thaw boar semen is deposited intra-utero, acceptable fertility (in terms of farrowing rates and litter size) is obtained. Currently, the most effective application of FT-semen for AI is achieved using deep uterine-AI (DUI) which allows placement of a minimal semen dose (in volume 0.5 to 10 mL and sperm number 0.5 to 1 x 10(9) total spermatozoa) into the anterior 1/3 of one uterine horn, with levels of fertility close to AI with liquid semen. However, owing to their shorter life span, FT-boar spermatozoa require an AI-to-ovulation interval not longer than 4-6 h, making peri-ovulatory AI a pre-requisite to obtain the highest possible fertility. Spontaneous ovulation most often occurs when two-thirds of oestrus has passed. Estimation of the duration of oestrus, taking into account the weaning-to-oestrus interval, is helpful when establishing appropriate AI-schedules. However, as the length of oestrus varies within and between farms, different AI strategies should be established a priori. The development of bio-sensors for spontaneous ovulation will widen the use of AI with frozen-thawed frozen semen.

Apatite nucleation on silica surface: a zeta potential approach.

Zeta potential measurements on pure silica, prepared by the sol-gel method from tetraethoxysilane under acidic conditions, are reported in different suspensions. Water suspensions and suspensions containing calcium or phosphate ions with and without NaCl were tested. zeta potential measurements were carried out as a function of the pH and ion concentration. Also, calcium and phosphate adsorption on silica was determined experimentally. The results of zeta potential and adsorption measurements suggest that both calcium and phosphate ions can be adsorbed on the silica surface; however, calcium adsorption is stronger than phosphate adsorption. When calcium and sodium ions are present in the suspension, calcium adsorption decreases. It seems that certain sites on the silica surface are specific for calcium adsorption.