Factors affecting fertility in frozen-thawed boar sperm

Frozen-thawed boar sperm holds the potential to have an impact on the future of the swine industry. Utilization of this technology could improve a swine producer’s ability to access top-tier genetics from around the world, to improve efficiency, profitability, and the quality of product to meet consumer demands. Effective application of frozen-thawed sperm can help reduce the potential risk associated with devastating economic loss due to the spread of disease. Frozen storage of boar sperm also provides a safeguard in the event of disease outbreaks, as genetic material from paternal lines can be preserved and banked for repopulation purposes. Historically these benefits have been masked by reduction in fertility measures such as litter size. The reduced fertility results from the damage sustained by the sperm cell during cryopreservation. However, increased understanding of this damage has lead to improved cryopreservation methods, ultimately increasing post-thaw viability and fertility. Enhancements in breeding technology have also resulted in a better understanding of the AI methods required to achieve acceptable farrowing rates and litter size. Fertility following AI with frozen-thawed sperm is approaching that of liquid stored sperm, and producers may soon reap the benefits of this technology. This thesis will outline the current swine industry, opportunities for utilizing frozen-thawed sperm, the main components of sperm, why they are susceptible to damage, and current freezing and breeding practices. Objective 1 was to develop a cryopreservation protocol for our lab that resulted in consistent post-thaw motility ( ≥ 40%) that would eventually be used by Illinois boar studs for domestic and international sale of frozen sperm. Evaluation with both manual microscopy and CASA methods were conducted to verify quality. A preliminary breeding trial was then conducted to test the fertility of sperm frozen with this method. There were 41 ejaculates from 23 boars used for freezing. Sperm were frozen at 1.4x109 sperm/mL, averaging 55.61.1% (meanSE) motility, following thaw. The samples assessed were not different (P>0.05) in motility when compared with manual or CASA systems, and results were most reliable at a 1:40 sperm dilution. In the preliminary breeding trial, gilts (n=14) were inseminated with either a single (n=10) or double (n=4) AI using 1, 2, or 4x109 motile, frozen-thawed sperm. Overall, the resulting pregnancy rates averaged 71.4% and numbers of normal fetuses per litter averaged 15.51.3 per litter. A feasibility study for freezing cost per ejaculate was estimated at $275/ejaculate or $11/dose of frozen-thawed semen at standard doses of 5x109 total frozen-thawed sperm. This cost estimate did not include genetic value, fixed equipment costs, depreciation, or variable lab space fees. Objective 2 focused on the proper methods for breeding with frozen-thawed boar sperm to achieve fertility. Our hypothesis was that increased numbers of inseminations and increased numbers of motile frozen-thawed sperm would improve pregnancy rate and litter size. Results showed acceptable fertility at high sperm numbers, but also the optimal method for insemination with the lowest dose tested. Gilts (n=111) responded to synchronization methods and were bred with 1, 2, or 4x109 motile frozen-thawed sperm from six boars using a single AI at 32 h, or a double AI, with the first AI at 24 and 32 h following estrus. Ultrasound was conducted at 12 h intervals to estimate the time of ovulation. On day 32 of gestation, overall pregnancy rate (73%) and number of normal fetuses per litter (10.80.5) across all treatments did not differ, and were not affected by number of motile sperm, or the interaction of number of motile sperm and number of inseminations. However, the number of inseminations tended to affect (P=0.14) the number of normal fetuses. Litter size increased with a double AI compared to single AI. Multiple inseminations helped to allow insemination to occur close to ovulation in response to variation in the time of ovulation. Both pregnancy rate and number of normal fetuses were greater when the time of the AI at 32 h occurred closer to the estimated time of ovulation (P<0.05). In addition, other factors such as presence of an abnormal ovary at day 30 decreased (P<0.001) pregnancy rate, while boar affected number of normal fetuses (P<0.01). Analysis of our data using a fertility index revealed doses of 2x109 motile sperm with multiple AI can achieve acceptable fertility with use of less sperm, when compared to AI using 4x109 motile sperm. The methods described here will investigate the potential for improved fertility when using frozen-thawed sperm, while accounting for variation in time of ovulation.

A pilot study on the productivity and behavior of the gestating sow housed in a flex stall

Animal welfare is a controversial topic in modern animal agriculture, partly because it generates interest from both the scientific community and the general public. The housing of gestating sows, particularly individual housing, is one of the most critical concerns in farm animal welfare. We hypothesize that the physical size of the standard gestation stall may limit movement and evoke demands and challenges on the sow to affect the physiological and psychological well-being of the individually housed sow. Thus, improvements in the design of the individual gestation stall system that allow more freedom to move, such as increasing stall width or designing a stall that could accommodate the changing size of the pregnant sow, may improve sow welfare. The objective of this pilot study was to evaluate the effects of a width adjustable stall (FLEX) on productivity and behavior of dry sows. The experiment consisted of 3 replications (block 1, n=4 sows; block 2, n=4 sows; block 3, n=8 sows), and multi-parious sows were allotted to either a FLEX stall or standard gestation stall for 1 gestation period. Sow mid-girth (top of the back to bottom of the udder) was measured 5-6 times throughout gestation to determine the best time points for FLEX stall width expansions. FLEX stall width was adjusted according to mid-girth measurements, and expanded to achieve an additional 2 cm of space between the bottom of the sow’s udder and floor of the stall so that sows could lie in full lateral recumbency without touching the sides of the stall. Productivity data recorded included: sow body weight (BW) and BW gain, number of piglets born and born alive, proportions of piglets stillborn, mummified, lost between birth and weaning, and weaned, and litter and mean piglet birth BW, weaning BW, and average BW gain from birth-to-weaning. Lesions were recorded on d 21 and d 111 of gestation. Sub-pilot behavior data were observed and registered for replicate 1 sows using continuous video-records for the l2 hour lights on period (period 1, 0600-1000; period 2, 1000-1400; period 3, 1400-1800) prior FLEX stall adjustment and 12 hour lights on period post adjustment on d 21, 22, 23, 43, 44, 45, 93, 94, 95. A randomized complete block design with a 2 × 2 factorial arrangement for treatments was used to analyze sow productivity and performance traits. Data were analyzed using the Mixed Models procedure of SAS. A preliminary analysis of data means and numerical trends was used to analyze sow behavior measurements. Sows housed in a FLEX stall had more (P < 0.05) total born and a tendency for more piglets born alive (P = 0.06) than sows housed in a standard stall. Sow body weight also tended to be higher (P = 0.06) for sows housed in a FLEX stall compared to sows housed in a standard stall. There were numerical trends for mean durations of sit, lay, lay (OUT), and eat behaviors to be greater for sows housed in a FLEX stall compared with sows housed in a standard stall. The mean duration of lay (IN) behavior tended to be numerically less for sows housed in a FLEX stall compared with sows housed in a standard stall. There were numerical trends for the mean durations of stand and drink behaviors to be greater for sows housed in a standard stall compared with sows housed in a FLEX stall. The mean frequencies of postural changes and mean durations of oral-nasal-facial and sham-chew behaviors were numerically similar between types of gestation stall. Mean durations and numerical trends indicate that time of day influenced all of the behaviors assessed in this study. The results of this pilot study indicate that the adjustable FLEX stall may affect sow productivity and behavior differently than the standard gestation stall, and thus potentially improve sow well-being. Future research should continue to compare the new FLEX stall design to current housing systems in use and examine physiological traits and immune status in addition to behavioral and productivity traits to assess the effects that this housing system has on the overall welfare of the gestating sow.