Relationship between sperm quality traits and field-fertility of porcine semen

An investigation involving seven boars, active in artificial insemination, and 1,350 multiparous sows was conducted at a private farm and aimed at examining the relationship between sperm quality traits and boar fertility in terms of farrowing rate and litter size. This experiment was done for 6 months. The semen samples were evaluated for subjective sperm motility and concentration. Ejaculates with at least 1 × 10⁸ sperm/mL and 70% sperm progressive motility were extended with a commercial medium to 30 × 10⁶ sperm/mL and used for artificial insemination (AI). AI dose was 100 mL semen containing 3 × 10⁹ spermatozoa. Aliquots of diluted semen were assessed for live morphologically normal spermatozoa (LMNS, eosin-nigrosin stain exclusion assay) and sperm chromatin instability (SCI, acridine orange assay). Farrowing rates according to different boar sperm varied (p < 0.001) from 59.3 to 88.92%. The mean values of LMNS (47.2~76.5%) and SCI (0.16~4.67%) differed significantly among boars. LMNS (r = 0.79, p < 0.05) and SCI (r = -0.90, p < 0.02) accounted for 62.2 and 81.7% of the variability in farrowing rates, respectively. After the combination of sperm traits, the relationship between percentage of LMNS with stable chromatin structure and farrowing rate was significant (r = 0.86, p < 0.05). The number of live piglets per parturition was not significantly correlated with sperm quality attributes. In conclusion, boar fertility after AI with freshly diluted semen can be predicted based on the evaluation of sperm morphology and chromatin integrity.     

Low Dose Insemination in the Sow – A Review

Artificial insemination (AI) in pigs has been established for about four decades but ejaculates are still used insufficiently. Higher demand of semen for AI and new techniques that involve low sperm concentration require the optimization of insemination protocols. Based on the knowledge of the physiology of sperm transportation and events in the female genital tract prior to fertilization, new strategies are under development to minimize sperm losses. One goal is to deposit the semen into the uterine horn rather than into the proximal cervix. It was shown that the minimal number of spermatozoa necessary for surgical AI at the utero‐tubal junction (UTJ) were at least 1 × 10⁶ diluted in 0.5 ml of a special extender. Artificial insemination into the distal part of the uterine horn required about 1 × 10⁷ million sperm in 20 ml of extender. Meanwhile, first insemination devices for non‐surgical intra‐uterine AI are commercially available. Using similar sperm concentrations as for surgical AI, non‐surgical uterine insemination did not differ significantly from control inseminations in terms of pregnancy rate and litter size. With respect to the fertilizing capacities of their ejaculates, boars have to be selected more strictly for sperm quality parameters as most of the compensatory effects of sperm cells disappear in maximally extended semen samples.     

Birth of puppies after intrauterine and intratubal insemination with frozen-thawed canine semen

The present study was performed to assess the fertility of frozen-thawed dog semen prepared by freezing with 6% glycerol and thawing at 70℃ for 8 sec, and to evaluate the least number of post-thaw spermatozoa necessary to achieve pregnancy by intrauterine or intratubal artificial insemination. It was found that the pregnancy rate of intrauterine artificial insemination was 100% using 6% glycerol buffer and thawing at 70℃ for 8 sec with 5 × 10⁷ spermatozoa. Even though the pregnancy rate (80%) and the whelping rate (24.5%) in the 5 × 10⁶ spermatozoa inseminated group were lower than those of the 5 × 10⁷ spermatozoa group, conception was confirmed with 5 × 10⁶ spermatozoa. Although the pregnancy rate of intratubal insemination was low (20%) with 4 × 10⁶ spermatozoa, this study is the first report to show the pregnancy rate of intratubal insemination with frozen-thawed ejaculated canine semen. In order to improve the pregnancy rate with intratubal insemination of canine spermatozoa, it is necessary to investigate the optimal insemination site of the uterine tube, the appropriate number of sperm, and the direct effect of buffer on oocytes.     

Bestimmung der osmotischen Resistenz von Eberspermien und deren Beziehungen zur Konservierungsfähigkeit von Samenproben

Contents Determination of the osmotic resistance of boar spermatozoa and the relations to sperm freezing and long storage The osmotic behaviour of fresh boar spermatozoa (482 ejaculates from 34 A.I. boars) was examined in a standard medium but with different osmotic pressures ranging from 150 mosm/kg till 500 mosm/kg. The percentage of spermatozoa with normal acrosomal ridge (NAR) was determined after incubation of 0.2 ml fresh semen in 3 ml of the osmotic test solutions at 39° C for 15 and 120 min. In the non-isotonic media % NAR decreased and could be quantified depending to incubation time and to the different osmolalities; NAR decreased strongest at 150 or 500 mosm/kg and after 120 min incubation time. Differences in % NAR were large between ejaculates and between boars but low within boars and statistically significant correlations had been found to % NAR and motility in froren/thawed semen samples or stored samples. Still higher correlations were found by calculation of an osmotic resistance test-value (ORT-value) which is defined as follows: ORT = 1/2 % NAR in isotonic medium/(after 15 min) +% NAR in 150 or 500 mosm/kg/(after 15 or 120 min) ORT-values can be calculated for hypotonic (150 mosm/kg) as well as for hypertonic (500 mosm/kg) test solutions and for 15 and 120 minutes incubation time. The hypotonic ORT-value for 120 min. is the most accurate; the values varied from 34 till 83 between ejaculates and from 44 till 76 between boars with a low variability within boars. Grouping of boars by their ORT-values in “good”, “moderate” and “poor” corresponded to a similar grouping of these boars by their semen quality (% NAR and motility) in preserved semen samples. Also a grouping of single ejaculates by their ORT-values corresponded to a similar qualitative classification of these ejaculates. ORT seems to become a good and practicable assessment test for the selection of boars suitable dor deep freezing or long storage of their spermatozoa. On the other hand preservation techniques too can be examined by ORT. But more investigations with this test must be done especially its relation to the fertilization capacity of semen samples with different ORT-values.     

Sperm localization in the oviducts of artificially inseminated dairy cattle

Eight animals, 3 heifers and 5 primiparous cows, were artificially inseminated by intrauterine deposition of frozen-thawed semen. The insemination dose comprised 20×10⁶ or 200 × 10⁶ spermatozoa, frozen in French mini straws. Four animals were inseminated at fixed time interval (72 or 84 h) after cloprostenol injection. The remaining 4 animals were inseminated in spontaneous oestrus. Slaughter took place 2 or 12 h after insemination. After fixation the oviducts were cut into segments, which were serial-sectioned and stained. Six sections per segment were examined under the microscope for sperm recovery.The number of spermatozoa recovered from the oviducts varied considerably among animals. Recovery was poor (less than 50 spermatozoa) in 4 animals. Recovery was low when insemination took place in induced oestrus and with the lower sperm number (20×10⁶). In animals in which more than 50 spermatozoa were found the distribution varied both between animals and between oviducts within the same animal. Overall, more spermatozoa were found in the lower (UTJ, isthmus and AIJ) than in the upper (ampulla) parts of the oviducts. In 3 out of 4 animals more spermatozoa were recovered from the left than from the right oviduct. Only in 1 animal were the majority of spermatozoa found in the oviduct ipsilateral to the follicle-bearing ovary.     

Efficiency of three boar sperm enrichment techniques

The objective of the study was to investigate the efficiency of three enrichment methods to separate boar spermatozoa. Twenty-four ejaculates from 12 boars (2 ejaculates/boar) were extended (30 × 10⁶ spermatozoa/mL) in commercial Beltsville Thawing Solution. Each semen sample was processed with glass wool column (GW) and glass beads (GB) filtration and with the single-layer centrifugation (SLC) technique. Semen samples before (control; C) and after treatment were evaluated for sperm CASA motility/kinetics and concentration, viability, morphology and chromatin integrity. Data were analysed with mixed models. The concentration of total and motile spermatozoa was significantly decreased after treatment in groups GW and SLC, but not in group GB. Group GW showed increased values of WOB compared with both groups C and GB. Group GB showed greater values of rapid movement spermatozoa and lower values of slow movement spermatozoa compared with group C. In group SLC, higher values of VSL, LIN and STR were observed compared with group C. In conclusion, all techniques under examination enhanced various CASA variables. Based on our results, the GB method is a promising alternative separation technique for boar sperm and deserves further research regarding swine in vitro fertilization.     

Fertility of deep frozen boar spermatozoa at various intervals between insemination and induced ovulation: influence of boars and thawing diluents.

The fertilizing ability of deep frozen boar spermatozoa was assessed after artificial insemination at various intervals after human chorionic gonadotropin (HCG) induced ovulation in gilts. Boar seminal plasma and OLEP were used as thawing diluent agents. The fertilization rates were similar when insemination was performed 2 or 6 hours before the time of estimated ovulation (40 hours post-HCG administration). However, fertility rates markedly declined when insemination was performed earlier than 6 hours before expected ovulation. In inseminations performed 2 hours before ovulation, the fertility rate was higher with specimens treated with boar seminal plasma diluent than with OLEP. As the difference between the time of insemination and that of ovulation increased, the difference in fertilizing ability of spermatozoa from the 2 boars used also increased. The results show that spermatozoa which have a low resistance to freezing-thawing also have a diminished fertile life in the female genital tract.     

Colloid centrifugation of boar semen

Colloid centrifugation of boar semen has been reported sporadically for at least the last two decades, beginning with density gradient centrifugation (DGC) and progressing more recently to single layer centrifugation (SLC). Single layer centrifugation through a species-specific colloid has been shown to be effective in selecting the best spermatozoa (spermatozoa with good motility and normal morphology) from boar sperm samples. The method is easier to use and less time-consuming than DGC and has been scaled-up to allow whole ejaculates from other species, e.g. stallions, to be processed in a practical manner. The SLC technique is described, and various scale-up versions are presented. The potential applications for SLC in boar semen preservation are as follows: to improve sperm quality in artificial insemination (AI) doses for 'problem' boars; to increase the shelf-life of normal stored sperm samples, either by processing the fresh semen before preparing AI doses or by processing the stored semen dose to extract the best spermatozoa; to remove pathogens (viruses, bacteria), thus improving biosecurity of semen doses and potentially reducing the use of antibiotics; to improve cryosurvival by removing dead and dying spermatozoa prior to cryopreservation; to select spermatozoa for in vitro fertilization. These applications are discussed and practical examples are provided. Finally, a few thoughts about the economic value of the technique to the boar semen industry are presented.     

Selenium Status and GSH-Px Activity in Semen and Blood of Boars at Different Ages Used for Artificial Insemination

This study was performed to determine the relationship between selenium (Se) content and Se-dependent glutathione peroxidase (GSH-Px) activity in blood and semen, and semen quality of boars at different age used in an artificial insemination (AI) station. Routine macroscopic and microscopic analyses of semen quality were accompanied by measurement of Se content and GSH-Px activity in blood and semen. The Se concentration in blood plasma, seminal fluid and spermatozoa was measured by fluorometric method, the GSH-Px activity by a method based on NADPH-coupled reaction. A total of 155 ejaculates and 58 blood samples were investigated. The results of this study showed that there was no direct relationship between the Se content and GSH-Px activity in blood plasma and semen, and semen quality of sexual matured boars. The mechanisms controlling Se content and GSH-Px activity in blood and semen seem to be independent. The age of boars as a differentiating factor for Se content and GSH-Px activity in blood and semen is possible. It is concluded that a determination of Se status and/or GSH-Px activity in organism before Se supplementation is indicated.     

Distribution of spermatozoa in the genital tract of artificially inseminated heifers

Eight heifers were artificially inseminated in the uterine body with 160×10⁶ spermatozoa frozen in French mini-straws. The heifers were slaughtered 2 (n = 4) or 12 (n = 4) h after insemination and spermatozoa were recovered by flushing defined segments of the reproductive tract. The efficiency of the method was checked in different ways. There was a slight underestimation of the number of recovered spermatozoa. This underestimation was randomly distributed among heifers and genital tract segments.The total number of spermatozoa recovered was higher at 2 than at 12 h (14.6 vs 0.6 % of the total number inseminated). Most spermatozoa were found in the vagina both at 2 and 12 h after insemination and in greater number at 2 h. In uterus there was a slight decline in the number of spermatozoa recovered at 2 versus 12 h after insemination. The number of spermatozoa recovered from the oviducts were similar at 2 (89.6 × 10³) and 12 h (71.5 × 10³) after insemination. At 2 h spermatozoa were found in all parts of the oviduct with the majority located in the utero tubal junction, whereas at 12 h the most were recovered from isthmus.More spermatozoa were recovered from the left than from the right side of the tract in 6 of the 8 heifers. Only in 1 heifer were the majority of spermatozoa found in the oviduct ipsilateral to the follicle bearing ovary.     

Freezability of equine semen after glass beads column separation

REASONS FOR PERFORMING STUDY: The success rate of artificial insemination following the freezing of stallion semen is limited; therefore, improving the stallion semen quality after the freezing and thawing process is a necessary objective. OBJECTIVES: To investigate the influence of glass bead column separation on the freezability of stallion semen. HYPOTHESIS: Glass beads in a column separator remove damaged and dead spermatozoa in the ejaculate during centrifugation. METHODS: In total, 50 ejaculates from 6 Lipizzaner stallions were studied. Each ejaculate was divided into 2 parts, one half processed following standard procedure and the second half used for the column separation procedure. After freezing, semen quality was evaluated using standard tests for motility, morphology and viability of semen. RESULTS: Motility and progressive motility of the column-separated (CS) semen were significantly higher (P < 0.001) before freezing and immediately, 24 and 48 h after thawing. A significant increase (P < 0.001) in the percentage of hypoosmotic positive spermatozoa was observed in CS samples. The percentage of total morphological changes in the separated samples before and after freezing was significantly lower (P < 0.001) compared with samples prepared using the standard procedure. A substantial decrease (P < 0.001) was found in the percentage of spermatozoa with damaged acrosomes. However, the percentage of spermatozoa with coiled tails was increased in the separated samples (P < 0.001). CONCLUSIONS: Column separation before freezing has a positive effect on the quality of thawed equine semen. POTENTIAL RELEVANCE: The quality of CS frozen/thawed samples indicates their potential use for increasing insemination success in mares.     

Porcine Field Fertility with Two Different Insemination Doses and the Effect of Sperm Morphology

In swine artificial insemination, several dose regimens are applied, ranging from 1.5 x 10(9) to 6.0 x 10(9) spermatozoa per intra-cervical insemination dose. A lower sperm dose is more profitable for artificial insemination centres and offers a more effective use of superior boars. To evaluate fertility, 50 boars were used for a total of 10 773 homospermic first inseminations at a dose of 2 billion spermatozoa. In addition, 96 boars were used at a dose of 3 billion spermatozoa for 34 789 homospermic first inseminations. Fertility was determined by a 60-day non-return rate (NR%) of first inseminations. Litter size was registered by total number of piglets born separately in primiparous and multiparous farrowings. On average, a sow was inseminated 1.5 times. A significant decrease was observed in all three fertility parameters (NR%, litter size of both primiparous and multiparous farrowings) with a dose of 2 billion spermatozoa compared with a dose of 3 billion spermatozoa. The NR% was 75.8% and 84.0% (p < 0.001), the mean litter size of primiparous farrowings 10.1 and 10.7 (p < 0.001) and the mean litter size of multiparous farrowings 11.7 and 12.1 (p < 0.001) for 2 and 3 billion spermatozoa/dose, respectively. The proportion of normal spermatozoa in the sperm morphology analysis correlated significantly with NR% in both insemination regimens: p < 0.001, r = 0.604 and p < 0.05, r = 0.223 for 2 and 3 billion spermatozoa/dose, respectively. These results confirm that quantity can at least partly compensate for poor sperm quality. When the boars with <70% normal spermatozoa in the morphology evaluation were excluded from the data there were no correlation between the sperm morphology and NR%. However, the difference between the NR% and litter size remained statistically significant (p < 0.001) in favour for the bigger insemination dose. In conclusion, a decrease in sperm dose from 3 to 2 billion spermatozoa on commercial farms will severely decrease prolificacy at least under field conditions, where a sow is inseminated an average of 1.5 times/heat, and the semen is typically used within 3 days after collection. We recommend that under commercial circumstances the homospermic semen doses contain no <3 billion spermatozoa/dose.     

The effect of photoperiod on selected parameters of boar semen

The objective of the present research was to determine the effect of increasing and decreasing natural photoperiods on selected parameters of boar ejaculates. The study material consisted of 17 boars: six Polish Large White (PLW) breed, five Polish Landrace (PL) breed, and six Duroc×Pietrain (D×P) crossbreed, all aged between 8 and 12 months at the beginning of the research. Analyses were conducted on 612 ejaculates, which were collected in two experimental periods: an increasing photoperiod (IP) (January–June) and a decreasing photoperiod (DP) (July–December). A statistically proven impact of photoperiod on the volume of semen was observed in all the studied breeds (P=0.004). During the decreasing photoperiod the mean volume of semen was 261.16±75.20 ml and this was almost 17 ml higher than that for the increasing period. For boars involved in the experiment, day length also had a significant impact on the total number of motile spermatozoa (P=0.037). In the increasing photoperiod the mean number was 3.26×10⁹ lower. A decreasing photoperiod has a positive affect on both boars and the parameters of the collected ejaculates, which were observed in the higher number of insemination doses per ejaculate. Between the different breeds, the reactions of boars to photoperiod differed and the most significant influence of photoperiod on semen parameters was noted among D×P breed boars. Least susceptible to changes in day length were PLW breed boars.     

Does a Boar's Season of Birth Determine Semen Parameters and Reproductive Performance?

This article studies the effect of a boar's birth season and breed on semen parameters and its further reproductive performance. Research material consisted of 72 boars from three breeds (24 Polish Large White PLW, 24 Polish Landrace PL, 24 Duroc × Pietrain D×P). During the whole period of the study, selected semen parameters were analysed: semen volume, spermatozoa concentration, total number of spermatozoa, total number of motile spermatozoa, number of insemination doses and also reproductive indicators: farrowing rate, total born litter size, total number of piglets born live and still, and average piglet weight. Boars born in the winter and summer months demonstrated the highest spermatozoa concentrations (383.25 and 392.37 × 10⁶/ml), total number of spermatozoa (91.75 and 93.21 × 10⁹), total number of motile spermatozoa (76.10 and 77.99 × 10⁹) and number of insemination doses (24.53 and 24.89; p ≤0.01). Statistically lower values for these parameters were observed for boars born in the spring and especially in autumn (p ≤0.01). The significant impact of birth season on farrowing rate (p ≤ 0.05) and average piglet weight (p ≤ 0.05) was confirmed for PLW boars. For the PL breed, only the total number of piglets born live was proven to be significantly affected (p ≤ 0.05). No impact of birth season was shown on semen quality or reproductive performance for D×P boars. In our study, we showed that the birth season of a boar had a more impact on the level of semen parameters, and less on the reproductive performance indicators. The results indicated that both the quality of semen and reproductive performance varied in terms of the study factors, as well as between individual breeds of boars involved in the experiment.     

Factors influencing boar sperm cryosurvival

Optimal sperm cryopreservation is a prerequisite for the sustainable commercial application of frozen-thawed boar semen for AI. Three experiments were performed to identify factors influencing variability of postthaw sperm survival among 464 boar ejaculates. Sperm-rich ejaculate fractions were cryopre-served using a standard freezing-thawing procedure for 0.5-mL plastic straws and computer-controlled freezing equipment. Postthaw sperm motility (assessed with a computer-assisted semen analysis system) and viability (simultaneously probed by flow cytometry analysis after triple-fluorescent stain), evaluated 30 and 150 min postthaw, were used to estimate the success of cryopreservation. In the first experiment, 168 unselected ejaculates (1 ejaculate/boar), from boars of 6 breeds with a wide age range (8 to 48 mo), were cryopreserved over a 12-mo period to evaluate the predictive value of boar (breed and age), semen collection, transport variables (season of ejaculate collection, interval between collections, and ejaculate temperature exposure), initial semen traits, and sperm quality before freezing on sperm survival after freezing-thawing. In Exp. 2, 4 ejaculates from each of 29 boars, preselected according to their initial semen traits and sperm quality before freezing, were collected and frozen over a 6-mo period to evaluate the influence of interboar and intraboar ejaculate variability in the survival of sperm after cryopreservation. In Exp. 3, 12 ejaculates preselected as for Exp. 2, from each of 15 boars with known good sperm cryosurvival, were collected and frozen over a 12-mo period to estimate the sustainability of sperm cryosurvival between ejaculates over time. Boar and semen collection and transport variables were not predictive of sperm cryosurvival among ejaculates. Initial semen traits and sperm quality variables observed before freezing explained 23.2 and 10.9%, respectively, of the variation in postthaw sperm motility and viability. However, more that 70% of total variance observed in postthaw sperm quality variables among ejaculates was explained by boar. This indicates that boar is the most important (P < 0.001) factor explaining the variability among ejaculates in sperm cryosurvival, with most (14 of the 15 boars in Exp. 3) showing consistent (P > 0.05) sperm cryosurvival over time.     

Influence of Boar Seminal Plasma on the Distribution of Spermatozoa in the Genital Tract of Gilts

The main purpose of the present study was to investigate whether boar seminal plasma affects the transport of spermatozoa in the genital tract of oestrous pigs or not, with special reference to the sperm transport into the oviducts. Altogether 17 gilts were used in three experiments.Experiment I. In nine gilts one uterine horn was injected surgically with 10¹⁰ spermatozoa suspended in seminal plasma and the other uterine horn with 10¹⁰ spermatozoa suspended in TESNaK-glucose buffer solution. The sperm deposition was performed under general anaesthesia. The gilts were slaughtered 1–2 or 4–6 h after insemination. The genital tract was removed and the numbers of spermatozoa determined in oviducts and in uterine horns.Experiment II. The insemination doses were prepared exactly as in Experiment I. Approx. 24 h before insemination Polyvinylchloride cannulas were inserted into the uterine lumen of the horns, drawn via the midventral incision at linea alba subcutaneously to cutaneous incisions ventral to the vulva opening. One cannula was placed in each uterine horn. At standing heat the insemination doses were slowly injected through the cannulas. The gilts were slaughtered 1 h after insemination and the numbers of spermatozoa within the genital tract were counted.Experiment III. In three gilts under general anaesthesia the uterine horns were ligated 10 cm from the uterotubal junction. The semen doses (containing 2 × 10⁹ spermatozoa), prepared as in Experiment I, were deposited into the uterine horns anterior to the ligatures through a cannula. The gilts were slaughtered 1 h after insemination, and the numbers of spermatozoa within the oviducts and ligated part of the uterine horns were counted.In all three experiments more spermatozoa were, on average, recovered in the oviducts connected to uterine horns inseminated with spermatozoa suspended in seminal plasma. In Experiments I andII this was the case for 10 of 14 gilts and in Experiment III for all the three gilts. It is therefore suggested that boar seminal plasma pro¬motes sperm transport into the oviduct of oestrous pigs. The back¬ground mechanism for this is discussed.     

Effect of collection frequency on quantitative and qualitative characteristics of pigeon (Columba livia) semen

The aim was to estimate the optimal frequency of semen collection from pigeons in relation to ejaculate volume, sperm concentration, total spermatozoa in ejaculate and percentage of live morphologically normal cells. The study was carried out on 455 ejaculates collected from two groups of pigeons, each of 10 males (group I: meat-type breed; group II: fancy pigeon). The birds were selected and kept individually in cages under a natural photoperiod. A two-person technique was used for semen collection (lumbo-sacral and cloacal region massage). Semen was collected once, twice or three times per week. Colour, consistency and volume of ejaculates were evaluated macroscopically immediately after collection. Sperm concentration and total number of cells in the ejaculate were estimated after dilution with Ringer's solution. A live-dead stain technique (nigrosin-eosin) was used to determine the percentage of live and normal spermatozoa. Semen collected 3x/week was of high quality. The average volume of a single ejaculate was small (21 microl in group I and 19 microl in group II), but sperm concentration was high--1.58 x 10(9)/ml and 1.96 x 10(9)/ml, respectively. The mean number of spermatozoa per ejaculate was 30.48 x 10(6) in group I and 39.49 x 10(6) in group II. An increased percentage of live and normal spermatozoa in semen collected more frequently was also observed. Collecting pigeon semen 3x/week provides spermatozoa in larger amounts and of better quality than less frequent collections (1x/week or 2x/week) and is recommended for obtaining more insemination doses.     

Fertility of Mares Inseminated With Frozen-Thawed Semen Processed by Single Layer Centrifugation Through a Colloid

The aim of this study was to determine whether there was an increase in pregnancy rates when frozen-thawed stallion semen was processed by single layer centrifugation (SLC) through a colloid before insemination. In addition, changes in semen parameters, including motility, were determined before and after SLC. Twenty light-horse mares (aged 3-16 years) and one Thoroughbred stallion (aged 16 years) having average fertility with fresh and cooled semen (>50% per cycle) and displaying a postthaw motility of >35% were used. Control mares were inseminated using 4- × 0.5-mL straws (200 × 10⁶/mL) of frozen-thawed semen. Treatment mares were inseminated with 4 × 0.5 mL of frozen-thawed semen after processing by SLC. Pregnancy rates were compared using Fisher exact test, and continuous parameters were evaluated by a Student t test. The pregnancy rates at day 14 were not different for the mares inseminated with control versus SLC-processed semen, despite the difference in sperm number (171 × 10⁶ ± 21, 59 × 10⁶ ± 25 progressively motile sperm). After frozen-thawed semen was processed by SLC, the percentage progressively motile sperm improved (P < .05), and SLC processing resulted in a 21.8% recovery of spermatozoa. In summary, centrifugation of frozen-thawed semen through a single layer of colloid increased the percentage of motile spermatozoa, but did not improve pregnancy rates after deep horn insemination.     

Distribution of Live and Dead Spermatozoa in the Genital Tract of Gilts at Different Times After Insemination

Twenty-four gilts were inseminated pair-wise with live or dead spermatozoa from the same ejaculate. The insemination dose was 100 ml undiluted semen containing, on average, 19×10⁹ spermatozoa. The gilts were slaughtered 1, 2, 6 and 12 h after insemination. The numbers of spermatozoa were counted in the uterus, uterotubal junction and in four equally long segments of the oviduct, called I–IV, with a haemocytometer. IV was adjacent to the uterotubal junction. The numbers of spermatozoa recovered in the uterus diminished significantly during the first 12 h after insemination. From gilts inseminated with live spermatozoa more spermatozoa were recovered in the uterotubal junction than from gilts inseminated with dead spermatozoa. Two h after insemination spermatozoa were recovered in all oviducts. Significantly more live than dead spermatozoa were recovered in Segments III and IV of the oviduct, regardless of time. In gilts inseminated with live spermatozoa the sperm count in Segment I varied significantly with time, being hiigest 2 h after insemination. At 6 and 12 h there were no distinct differences in the distribution of live spermatozoa between the various oviduct segments. The numbers of spermatozoa recovered in the oviduct were at these times apparently related to the sperm depots in the uterotubal junction.     

Influence of boars on the relationship between fertility and post thawing sperm quality of deep frozen boar spermatozoa.

The possibility of selecting boars for deep freezing of spermatozoa was evaluated by tests of frozen-thawed spermatozoa. 20 of 31 ejaculates were used for artificial insemination of 37 gilts. Boar seminal plasma and OLEP were used as thawing diluents. The thermoresistance test and the extracellular concentration of aspartate aminotransferase (ASAT) were used as indicators of fertility. Spermatozoa from 4 boars were utilized. 1 of the boars appeared to be of superior fertility, while the spermatozoa from another was infertile. The latter animal had been used for fresh artificial insemination trials, and had shown higher pregnancy rates than the average. Samples from this boar exhibited the lowest degree of motility after 3 hours storage at 37 degrees C, the greatest relative decrease of motility during the thermoresistance test, the greatest release of ASAT after thawing by OLEP, and greatest relative release of ASAT. These variations were statistically significant (p less than .05). The results show that there is a possibility of detecting boars with spermatozoa with potential low freezability.