Boar Fertility Research Articles

Logistic Regression Analysis Factors Affecting Sperm Motility and Abnormal Sperm Morphology in Boars.

Simple SummaryReduced sperm motility and morphological abnormalities have significant negative impacts on conception rates in sows and are important indexes of semen. The identification of factors that influence sperm motility and morphology will improve boar fertility in commercial herds. On the basis of analysis of 5042 ejaculates from 385 boars using a logistic regression model, we found that serum Cu excess, serum Fe deficiency, and Pb presence in seminal plasma were risk factors for poor semen quality in boars. More importantly, the presence of seminal plasma Pb had more serious effect on the probability of abnormal sperm morphology than serum Cu excess and serum Fe deficiency. In addition, Yorkshire and Landrace boars had higher sperm motility and lower abnormal sperm morphology than Duroc boars. The difference in serum and seminal plasma elements among boars with different semen qualities may become a guide for regulating these elements used in boar diet. The highly predictive values of serum Cu, Fe, and seminal plasma Pb could be used in the future as an additional tool in semen quality evaluation.Logistic regression models, including variables of boar breed, age, serum, and seminal plasma elements, were used to identify the influencing factors of sperm motility and morphology in this study. Sperm motility degree was classified as grade 0: ≤85% and grade 1: >85%. Abnormal sperm morphology was classified as grade 0: ≤10%, grade 1: 10–20%, and grade 2: >20%. Element concentration of 385 boars was detected by inductively coupled plasma mass spectrometry. Results showed that boars with serum Cu ≥ 2.5 mg/L had lower sperm motility (odds ratio (OR): 0.496; 95% confidence interval (CI): 0.285–0.864) and higher abnormal sperm morphology (OR: 2.003; 95% CI: 1.189–3.376) than those with serum Cu ≤ 2.0 mg/L. Boars with serum Fe ≥ 1.5 mg/L had lower abnormal sperm morphology than those with serum Fe ≤ 1.0 mg/L (OR: 0.463; 95% CI: 0.255–0.842). The presence of Pb in seminal plasma increased abnormal sperm morphology. The probability of abnormal sperm morphology >20% from boars with seminal plasma Pb increased with a range of 5.78–15.30% than that from boars without seminal plasma Pb among three breeds. In conclusion, serum Cu excess, serum Fe deficiency, and seminal plasma Pb are risk factors for poor semen quality in boars.

Porcine seminal protein-I and II mRNA expression in boar spermatozoa is significantly correlated with fertility

In recent years, genomic and proteomic biomarkers have been identified for the diagnosis of male fertility to overcome the limitations of conventional semen analysis. Owing to the limited genes available so far, the single gene approach is commonly adopted for analyzing the phenotype of interest. However, the single-gene approach is less effective than multiple-gene strategies for diagnosing a desirable phenotype. Herein, we investigate the ability of two fertility-related genomic markers (porcine seminal protein (PSP)-I and PSP-II) in spermatozoa to predict boar litter size in addition to conventional semen parameters. First, we examined different semen parameters (motility, motion kinematics, and capacitation status) and gene expression in high- and low-litter size boar spermatozoa. Then, we evaluated the correlation of these parameters with the fertility of 21 Yorkshire boars. Finally, we investigated the efficacy of single/combined markers to predict male fertility using a comprehensive statistical model. Our result showed that there were no significant differences in sperm motility, motion kinematics, or capacitation status, however, the mRNA expression of PSP-I and PSP-II in spermatozoa was significantly different in high- and low-litter size boars. In the individual screening test, the expression of both genes was negatively correlated with boar fertility (r = 0-0.578 and −0.456, respectively), whereas only hyperactivation (HYP) showed a positive correlation (r = 0.444) among the tested semen parameters. As single markers, PSP-I and PSP-II have a better diagnostic power to predict boar fertility, regardless of HYP, in quality assessment analyses. In addition, when these markers were combined, the positive predictive value, negative predictive value, and overall test effectiveness for fertility detection were improved. Surprisingly, when PSP-I and PSP-II were considered together, the deviation of the predicted average litter size between high- and low-litter size boars was 1.77. Based on the findings, we suggest that the use of genomic markers in spermatozoa rather than commonly analyzed semen parameters may be more accurate for evaluating male fertility. Moreover, using a combination of markers could increase the overall accuracy of (in)fertility predictions, and thus, could be considered for field application.

Cryopreservation Differentially Alters the Proteome of Epididymal and Ejaculated Pig Spermatozoa.

Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein–protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.

Boar seminal plasma proteins and their relevance to reproductive technologies

Seminal plasma proteins participate in a number of events important for fertilization and the establishment of pregnancy. As a result, attempts have been made to use them to enhance reproductive performance associated with several swine reproductive technologies. Inclusion of seminal plasma into cryopreservation and sex-sorting protocols improved sperm viability and membrane integrity and suppressed capacitation-like changes which are considered to be major challenges associated with these techniques. Unfortunately, it has yet to be shown that these improvements consistently increase in vivo fertility. In contrast, pre-breeding administration of seminal plasma in conjunction with conventional breeding regimens improved farrowing rates and numbers of pigs born alive on commercial farms that already had very good reproductive performance. The best way to capture these beneficial effects in A.I. programs currently is being investigated. Finally, three seminal plasma proteins appear to have reasonable correlations with fertility in boars that normally produce sperm with excellent motility and morphology. They hold potential for development of prospective male fertility tests. However, there is some evidence that indicates consideration of the complete profile of a boar’s seminal plasma proteins may be more appropriate for this purpose as opposed to concentrating on individual ones independently. Preliminary results from a field study indicate that farrowing rate and litter sizes are superior in boars with high levels of two seminal plasma proteins associated with fertility compared with their counterparts in which only one of these is elevated. All of these technologies will benefit from continued research efforts devoted to the additional characterization of proteins in seminal plasma and elucidation of their biological effects on swine reproductive physiology.

Sperm solute carrier family 9 regulator 1 is correlated with boar fertility

Predicting male fertility is extremely important for artificial insemination and profitable farm management. Conventional semen assessment together with computer-assisted sperm analysis is widely used to predict male fertility under field conditions. However, the clinical validation and sensitivity of these methods remain unclear. Therefore, a new approach is needed to predict male fertility. Here, we investigated the use of a transcriptomic marker (solute carrier family 9, subfamily A, member 3, regulator 1; SLC9A3R1) together with sperm motility parameters and capacitation status to predict fertility/infertility in boars at the commercial level. Our data showed that among motility parameters and the capacitation status, hyperactivation (HYP) differed between high- and low-litter size boars. HYP showed a significant positive correlation (R = 0.468) with boar litter size. Simultaneously, the expression of SLC9A3R1, a gene important in sperm ion channel regulation, was significantly negatively correlated (R = −0.523) with boar litter size. Quality assessment revealed that both HYP and SLC9A3R1 showed considerable sensitivity (71.43 vs. 100%), specificity (100 vs. 71.43%), and overall accuracy (90%) for predicting male fertility. Interestingly, the potential of SLC9A3R1 expression to increase the average piglet number per breeding was higher (0.7 piglets) than that of HYP (0.5 piglets). Thus, measuring SLC9A3R1 expression in spermatozoa may be a more accurate marker for evaluating male fertility/infertility than conventionally used motility parameters and capacitation status.

157 Dietary L-arginine supplementation affects boar seminal plasma proteome

There are numerous benefit claims for the supplementation of l-arginine in animal production systems. Its positive effect on fertility has been widely characterised in females of various species; however, the effects on the male reproductive system remain debatable with still unfolding molecular mechanisms. Here we investigated the effects of a dietary l-arginine supplementation on boar semen production outcomes, sperm characteristics, and seminal plasma proteome. Nine mature (20 months of age) Nebraska Index Line boars were individually housed and randomly assigned to a soybean meal-based diet containing 0.77% standard l-arginine (control, n=4) or 1.77% high l-arginine (n=5). Boar semen was collected weekly during the 6-week dietary arginine trial. Semen production outcomes (volume and total spermatozoa) and sperm motion (total and progressive motility) and morphology characteristics were subsequently analysed using a computer-assisted sperm analyzer (CEROS II). On Week 6, the seminal plasma of each boar was obtained by centrifugation at 4°C for proteome analysis. The clarified seminal plasma was precipitated; the purified proteins were resuspended in an appropriate buffer and loaded (300μg) onto an immobilized pH gradient strip (isoelectric point 3-10) for the first-dimension electrophoresis. The second-dimension was subsequently performed (4-20% gradient gel), and all gels were stained with Coomassie R250 dye. The PDQuest software (Bio-Rad, Hercules, CA, USA) was used to detect all significantly differentially expressed protein spots (Student’s t-test with P<0.05). Two-way ANOVA repeated measurements (SPSS Statistics, Chicago, IL, USA) was used to analyse dependent (semen outputs and sperm characteristics) and independent (week) variables. A difference was significant at P<0.05. During the l-arginine-consumption trial, semen volumes and total spermatozoa were not significantly affected (P>0.05). Similarly, various sperm motility (total and progressive motility) and velocity characteristics were not affected (P>0.05). By the end of the feed trial, the average (mean±standard error of the mean) semen volume (389±40mL), the total spermatozoa per ejaculate (39±8×109), the total (82±2%) and progressive (62±3%) sperm motility, the sperm velocity (e.g. average path: 98±7μm s−1), and sperm morphology (e.g. distal droplet: 5±1%) parameters of the control group were not significantly different from the group supplemented with high l-arginine: 393±35mL, 47±4×109, 82±2%, 61±3%, 92±8μm s−1, and 5±0%, respectively (P>0.05). In contrast, the proteome profiles of seminal plasma harvested on the sixth week of diet revealed various changes, characterised by the significantly increased expression of 8 protein spots in seminal plasma samples derived from arginine-fed boars (P<0.05). These results indicate that dietary supplementation of l-arginine does not affect the semen outputs, neither the sperm motility characteristics nor their morphology. However, the proteome profile of the seminal plasma was changed by the presence of l-arginine. The findings may have significant implications for boar fertility, and the identification of these proteins is ongoing. This work was supported by USDA-Agricultural Research Service Biophotonics Initiative #58-6402-3-018.

The Proteome of Pig Spermatozoa Is Remodeled During Ejaculation.

Proteins are essential for sperm function, including their fertilizing capacity. Pig spermatozoa, emitted in well-defined ejaculate fractions, vary in their functionality, which could be related to different sperm protein composition. This study aimed (i) to update the porcine sperm proteome and (ii) to identify proteins differentially expressed in mature spermatozoa from cauda epididymis and those delivered in separate ejaculate fractions. Ejaculates from nine mature and fertile boars were manually collected in three separate portions: the first 10 ml of the sperm-rich ejaculate fraction (SRF), the rest of the SRF and the post-SRF. The contents of cauda epididymides of the boars were collected post-mortem by retrograde duct perfusion, generating four different semen sources for each boar. Following centrifugation, the resulting pellets of each semen source were initially pooled and later split to generate two technical replicates per source. The resulting eight sperm samples (two per semen source) were subjected to iTRAQ-based 2D-LC-MS/MS for protein identification and quantification. A total of 1,723 proteins were identified (974 of Sus scrofa taxonomy) and 1,602 of them were also quantified (960 of Sus scrofa taxonomy). After an ANOVA test, 32 Sus scrofa proteins showed quantitative differences (p < 0.01) among semen sources, which was particularly relevant for sperm functionality in the post-SRF. The present study showed that the proteome of boar spermatozoa is remodeled during ejaculation involving proteins clearly implicated in sperm function. The findings provide valuable groundwork for further studies focused on identifying protein biomarkers of sperm fertility.

Effect of Breed on Frequency of Morphological Defects in Boar Spermatozoa

The aim of this study was to assess an effect of breed on frequency of morphological changes in boar spermatozoa. A total of 1080 semen samples were used in this study. The samples were collected from January to December 2017 from 90 boars of the Duroc, Czech Large White and Czech Landrace breeds. Sperm cell abnormalities were evaluated microscopically from dyed smears of native semen on glass slides. Defects of spermatozoa were divided according to their impact on fertility of boars into categories of “major defects” (with a great negative effect on fertility) and “minor defects” (with a mild effect on fertility). The overall frequency of spermatozoa with morphological defects ranged from 11.62 % in Czech Landrace to 20.96 % in Duroc. Statistically significant differences were found in the frequencies of both major and minor defects among the breeds. The results of this study document that there are great inter‑breed differences in the occurrence of morphologically defective spermatozoa that significantly affect fertility of boars.

Relationship between sperm motility characteristics and ATP concentrations, and association with fertility in two different pig breeds

Boar fertility has a major impact on overall pig reproductive efficiency. Using accurate and objective in vitro sperm variables for predicting in vivo fertility from a single ejaculate, however, is challenging. Motility is the most widely used indicator of sperm quality, and a computer assisted sperm analysis (CASA) system is now available for objective assessment of sperm motility characteristics. In this study sperm motility characteristics and semen ATP concentrations were investigated and the effect of both were evaluated on total number of piglets born (TNB) when Norwegian Landrace (NL) and Norwegian Duroc (ND) boar semen was used for AI. In addition, breed differences for semen storage capacity were investigated. The results from CASA analysis indicated there were differences between NL and ND sperm motility variables. The percentage of motile sperm cells decreased in both NL (P = 0.01) and ND (P < 0.0001) during storage. A large proportion of sperm cells with a hyperactive motility pattern were detected in ND semen on the day of collection, with no significant changes as a result of storage. Inconsistent with this finding, there was greater degree of hyper-activation in sperm motility pattern for NL because of semen storage. There was a significant decrease in semen ATP concentration during storage (P < 0.0001) in both breeds. The linearity of sperm movement at the day of collection and the wobble after storage influenced TNB in NL, while the percentage of motile cells, curvilinear velocity and lateral head amplitude on the day of semen collection and linearity after storage influenced TNB in ND.

PIG PRODUCTIVITY UNDER PURE BREEDING AND CROSSBREEDING

For the research the groups were formed by method of analogues based on origin, age, physiological state, live weight. Studying sperm productivity, the number of ejaculates obtained during the year, average volume, concentration, and the total number of spermatozoa in the ejaculate were taken into account and conducted according to conventional techniques using a digital photometer “SDM-30” produced by "Minitüb”. Indicators of economical useful traits of the test animals were calculated according to primary zoo-technical accounting by standard methods of biometric analysis. To determine breeding value of animals by the BLUP method, software developed at the Institute of Pig Breeding "System determining the breeding value of pigs" was used. In carrying out the work, population genetics and mathematical methods were applied for studying of patterns of variability, repeatability, assessment of the genotype of animals.&#x0D; Reproductive quality of the sows was assessed by multiparity (total number of piglets at farrowing), prolificacy (piglet’s weight at birth, kg), milk ability (total weight of litter at 21 days, kg) piglet’s weight at weaning and survival of offspring to weaning. Survival of piglets, homogeneity and uniformity of litter were determined by modified M. D. Berezovsky – D. V. Lomako index and V. P. Kovalenko homogeneity index. Growth and development were controlled by change of individual live weight via weighing. Average daily gain was determined based on the initial and final live weight and number of days between weightings.&#x0D; Dynamics of indicators of growth and development, maternal qualities of sows and sperm productivity and fertility of boars at the purebred animal of initial breeds and live weight and age at 100 kg in the hybrids of first generation were studied. The youngsters obtained at crossbreeding Large White breed of domestic and foreign selection with Landrace had the best indicators of growth and development than the purebred counterparts in all age periods. Higher live weight at weaning at age of 60 days was typical for combination ♀LWE x ♂LE and ♀LE x ♂LWE; it is quite natural, because they had lower average number of piglets at weaning. For maturing volumes of back of the carcass and a well-developed layer of fat should be adjusted to Landrace sows and Large White boars. Fattening of commercial hybrids obtained by this scheme under Ukrainian conditions is advisable to check out at animal’s live weight of 115 ... 120 kg.&#x0D; Significant correlations were proven on the basis of: BLUP index – back fat thickness at the level of the 6-7 thoracic vertebra – -0,221 ± 0,0938 (tr = 2,35), BLUP index – back fat thickness on sacrum – -0,298 ± 0,0898 (tr = 3,31), BLUP index – back fat thickness at the midpoint of the back between the withers and sacrum – -0,239 ± 0,0929 (tr = 2,57), BLUP index – body length – -0,338 ± 0,0873 (tr = 3, 86), BLUP index – multiparity – -0,294 ± 0,0900 (tr = 3,26), BLUP index – total weight of the litter at the date of weaning – -0,233 ± 0,0932 (tr = 2,49). Keeping the selection process towards selection and combination of parental pairs by BLUP method is one of the ways to increase the productive capacity of the animals.&#x0D; According to the research it was found that asymmetry of testes sizes was observed together with individual peculiarities of testes form at the same breeding boars. Testes have physiological asymmetry. Ratio of areas of the left testes to right was Sl/Sr = 1.04, and volumes respectively Vl/Vr = 1.20, on average, left testes by their areas were 2.7% more than the right, and by volumes – 14.6% respectively.&#x0D; Comparing the live weight of boars with weight of their testes does not have a logical pattern. The average weight of testes was 0.14% of the live weight of breeding boars. Total volume of ejaculate on a group of boars was 355,3 ± 16,9 ml. The concentration of sperm in the ejaculate was 64,2 ± 4,6 billion, activity – 8,7 ± 0,2 points. The studies found that boars with greater weight of testes produce sperm with more sperm concentration and that's why more sperms in the ejaculate were obtained from them.&#x0D; Combination of Landrace and Large White breeds is apposite for obtaining precocious pigs with well-developed layer of fat at the 6-7 thoracic vertebra and high level of prolificacy (1,8-1,9 kg).

New In-Depth Analytical Approach of the Porcine Seminal Plasma Proteome Reveals Potential Fertility Biomarkers.

A complete characterization of the proteome of seminal plasma (SP) is an essential step to understand how SP influences sperm function and fertility after artificial insemination (AI). The purpose of this study was to identify which among characterized proteins in boar SP were differently expressed among AI boars with significantly different fertility outcomes. A total of 872 SP proteins, 390 of them belonging specifically to Sus Scrofa taxonomy, were identified (Experiment 1) by using a novel proteomic approach that combined size exclusion chromatography and solid-phase extraction as prefractionation steps prior to Nano LC-ESI-MS/MS analysis. The SP proteomes of 26 boars showing significant differences in farrowing rate (n = 13) and litter size (n = 13) after the AI of 10 526 sows were further analyzed (Experiment 2). A total of 679 SP proteins were then quantified by the SWATH approach, where the penalized linear regression LASSO revealed differentially expressed SP proteins for farrowing rate (FURIN, AKR1B1, UBA1, PIN1, SPAM1, BLMH, SMPDL3A, KRT17, KRT10, TTC23, and AGT) and litter size (PN-1, THBS1, DSC1, and CAT). This study extended our knowledge of the SP proteome and revealed some SP proteins as potential biomarkers of fertility in AI boars.

RNA sequencing reveals candidate genes and polymorphisms related to sperm DNA integrity in testis tissue from boars

BackgroundSperm DNA is protected against fragmentation by a high degree of chromatin packaging. It has been demonstrated that proper chromatin packaging is important for boar fertility outcome. However, little is known about the molecular mechanisms underlying differences in sperm DNA fragmentation. Knowledge of sequence variation influencing this sperm parameter could be beneficial in selecting the best artificial insemination (AI) boars for commercial production. The aim of this study was to identify genes differentially expressed in testis tissue of Norwegian Landrace and Duroc boars, with high and low sperm DNA fragmentation index (DFI), using transcriptome sequencing.ResultsAltogether, 308 and 374 genes were found to display significant differences in expression level between high and low DFI in Landrace and Duroc boars, respectively. Of these genes, 71 were differentially expressed in both breeds. Gene ontology analysis revealed that significant terms in common for the two breeds included extracellular matrix, extracellular region and calcium ion binding. Moreover, different metabolic processes were enriched in Landrace and Duroc, whereas immune response terms were common in Landrace only. Variant detection identified putative polymorphisms in some of the differentially expressed genes. Validation showed that predicted high impact variants in RAMP2, GIMAP6 and three uncharacterized genes are particularly interesting for sperm DNA fragmentation in boars.ConclusionsWe identified differentially expressed genes between groups of boars with high and low sperm DFI, and functional annotation of these genes point towards important biochemical pathways. Moreover, variant detection identified putative polymorphisms in the differentially expressed genes. Our results provide valuable insights into the molecular network underlying DFI in pigs.

Nanotechnology-based selection of boar spermatozoa: growth development and health assessments of produced offspring

Abstract The heterogeneous population of spermatozoa within the semen ejaculate influences the sire fertility. The current design of magnetic nanoparticle conjugates allows for selective targeting and removal of non-viable spermatozoa within the semen ejaculate. However, the safe application of this process, termed as nanoselection or nanopurification in previous studies, in food animal production and toxicity concerns has yet to be explored. Here, we assessed the fertility potential of nanoselected boar spermatozoa and the subsequent post-natal growth and health characteristics of resulting offspring. Semen doses were harvested from three fertile boars (n=4 doses per boar) and split in two groups (2 doses/boar). Six semen doses (2 per boar)) were subjected to the targeted depletion of acrosome membrane damaged and apoptotic spermatozoa (nanoselected). Meanwhile, the remaining semen doses (n=6) were maintained in the shipping Styrofoam box (control). The motility characteristics of both control and nanoselected spermatozoa were evaluated before and after nanoselection, followed by their use for double inseminations of six estrus synchronized gilts (2 doses/boar/gilt; 3 gilts /control or nanoselected). In comparison to the controls, the computer-assisted sperm analyzer (Hamilton-Thorne) revealed greater motion characteristics of nanoselected spermatozoa, with a significantly higher proportion of progressive spermatozoa and straightness (P 0.05). Various developmental and health parameters of produced offspring such as hepatic cytochrome P450 enzyme activities, blood glucose and immunoglobulin G concentrations, hematocrit, and white blood cell proportions were similar between and across all pigs (control and nanoselected). In addition, reproductive tracts of females born from nanoselected spermatozoa showed no indication of impaired fertility potential, although a significant shortened uterine horn length was measured (56.3±2.6 cm vs. 64.4±2.2 cm in the control group, P=0.04). In conclusion, findings revealed no obvious perturbations of sperm function following nanoselection, while post-natal growth, development, or health data of derived offspring suggest absence of inflicted sub-lethal toxicities attributable to sperm nanoselection. This study supports the safe use of the proposed nanotechnology-based selection for effective semen handling in terminal line swine production systems.

Scrotal insulation and sperm production in the boar.

Seasonal infertility is a limiting factor in boar fertility, and is increasingly important as climate changes. Spermatogenesis in the boar produces 256 spermatozoa per type A1 spermatogonium, but the process is inefficient such that only 10-30% of these potential spermatozoa are actually produced. Heat further impacts spermatogenesis by reducing the number of specific germ cells produced while increasing the fraction of abnormal sperm. Early studies used whole-animal exposure to simulate seasonal exposure to heat under production settings, but this approach is associated with many confounding factors that make assessment of the mechanisms of heat-induced damage to spermatogenesis difficult. Scrotal insulation provides a better model to investigate the mechanisms and potential mitigation strategies of heat-induce damage. For example, scrotal insulation helped identify a link between short-term heat stress and damage to meiotic germ cells. This outcome is likely due to changes in the integrity of the blood-testis barrier, which induce apoptosis, autophagy and DNA damage in the germ cells. Further understanding how heat damages spermatogenesis, and whether or not this can be repaired, are crucial to mitigating heat effects on boars in production settings.

Active paraoxonase 1 is synthesised throughout the internal boar genital organs.

The paraoxonase type 1 (PON1) is an enzyme with antioxidant properties recently identified in the seminal plasma (SP) of several species, including the porcine. The aims of the present study were to (1) describe the immunohistochemical localisation of PON1 in the genital organs of fertile boars and (2) evaluate the relationship among PON1 activity and high-density lipoprotein cholesterol (HDL-C) concentration in fluids of the boar genital organs. Immunohistochemical analysis demonstrated that PON1 was present in testis (specifically in Leydig cells, blood vessels, spermatogonia and elongated spermatids), epididymis (specifically in the cytoplasm of the principal epithelial cells, luminal secretion and in the surrounding smooth muscle) and the lining epithelia of the accessory sexual glands (cytoplasmic location in the prostate and membranous in the seminal vesicle and bulbourethral glands). The Western blotting analysis confirmed the presence of PON1 in all boar genital organs, showing in all of them a band of 51 kDa and an extra band of 45 kDa only in seminal vesicles. PON1 showed higher activity levels in epididymal fluid than those in SP of the entire ejaculate or of specific ejaculate portions. A highly positive relationship between PON1 activity and HDL-C concentration was found in all genital fluids. In sum, all boar genital organs contributing to sperm-accompanying fluid/s were able to express PON1, whose activity in these genital fluids is highly dependent on the variable HDL-C concentration present.

Prediction of male fertility using capacitation‐associated proteins in spermatozoa

Infertility and subfertility account for huge economic losses in the animal industry; indeed, 50% of animal breeding failure is associated with male infertility. Approximately 70% of cattle and 90% of pig livestock are currently produced by artificial insemination. Therefore, breeding-male selection is extremely important for the genetic benefits of progeny. Although conventional semen analysis provides an initial measure of male fertility, its clinical value is questionable. Proteomics approaches recently identified candidate protein markers in spermatozoa for evaluating male fertility. Fertility-related proteins in capacitated boar spermatozoa were shown to predict boar fertility more precisely then those detected in ejaculated spermatozoa, which motivated the development of more accurate and sensitive tools for the assessment of male fertility in relation to sperm function and fertilization. Although protein markers in spermatozoa are capable of discriminating fertile and infertile males, clinical trials are required to validate their predictive utility. This review outlines recent findings regarding the capacitation-related proteome of spermatozoa, and discusses how these proteins may be utilized to better understand the fertility of domestic animals.

Fertilitas Spermatozoa Babi Landrace dalam Pengencer Modifikasi Zorlesco dengan Susu Kacang Kedelai

&lt;p&gt;A research has been done to study the effectiveness of Zorlesco extender and modified Zorlesco extender with soymilk to maintain sperm fertility of landrace boar preserved at 18˚C. Semen was collected twice a week by hand massage from 2 heads of ± 2.5 years old landrace boar. A Completely Randomized Design with 4 treatments and 5 replications was used to organized the experiment. The fresh evaluated semen were extended with zorlesco P(0), zorlesco + hand made soymilk P(1), zorlesco + MetabolisTM soymilk P(2), and zorlesco + MelileaTM soymilk P(3). Extended semen were preserved at 180C. Analysis of variance and Duncan test were used to analyzed data of motility and viability of sperm. The result showed that the modification of diluent Zorlesco added with 5% soymilk could not defend by sperm motility and viability spermatozoa compared with Zorlesco for 42 hours storage (P&amp;lt;0.05).&lt;br /&gt;Keyword: Soymilk, modified zorlesco, sperm motility, sperm viability, landrace boar&lt;/p&gt;

Fertilitas Spermatozoa Babi Landrace dalam Pengencer Modifikasi Zorlesco dengan Susu Kacang Kedelai

&lt;p&gt;A research has been done to study the effectiveness of Zorlesco extender and modified Zorlesco extender with soymilk to maintain sperm fertility of landrace boar preserved at 18˚C. Semen was collected twice a week by hand massage from 2 heads of ± 2.5 years old landrace boar. A Completely Randomized Design with 4 treatments and 5 replications was used to organized the experiment. The fresh evaluated semen were extended with zorlesco P(0), zorlesco + hand made soymilk P(1), zorlesco + MetabolisTM soymilk P(2), and zorlesco + MelileaTM soymilk P(3). Extended semen were preserved at 180C. Analysis of variance and Duncan test were used to analyzed data of motility and viability of sperm. The result showed that the modification of diluent Zorlesco added with 5% soymilk could not defend by sperm motility and viability spermatozoa compared with Zorlesco for 42 hours storage (P&amp;lt;0.05).&lt;br /&gt;Keyword: Soymilk, modified zorlesco, sperm motility, sperm viability, landrace boar&lt;/p&gt;

Fertility of boars - what is important to know

The most important part in reproductive management is the control of boar fertility. A common division of fertility traits is on the: in vitro (sperm traits) and in vivo (return rate, farrowing rate and litter size traits) fertility. In many studies were found differences between breed in the both groups of fertility traits. Variability of sperm traits of boars during the reproductive exploitation is influenced by various genetic (boar, breed) and paragenetic factors/effects (age, season, intensity of use). Good libido is desirable characteristics in boars, but the knowlegde of the correlation of libido and boar fertility traits are limited. Also, there is no standardised procedure or methods for the estimation of libido of the boars. The permanent ranking of boars according to the reproductive efficiency should be performing. Good reproductive management implies the timely identification of boars with the low fertility (or close to the average).

24 RELATIONSHIP BETWEEN BOAR SPERM TRAITS AND FERTILITY RATE FOLLOWING ARTIFICIAL INSEMINATION UNDER SMALLHOLDER PRODUCTION SYSTEMS

The prediction of sperm fertility has a great economic importance to the pig breeding industry. The objective of the study was to determine the relationship between boar sperm quality and fertility following artificial insemination (AI) under smallholder production systems. A total of 18 ejaculates were collected from 3 breeding boars using a hand-gloved technique. Aliquots of diluted semen were assessed for sperm motility using a computer aided sperm analysis before AI. Sperm viability was evaluated using Synthetic Binding CD-14 (SYBR-14+)/propidium iodide (PI–), whereas sperm morphology was evaluated using Eosin Nigrosin staining. Fluorescent microscope was used at 100× magnification to count 200 sperm per slide. The semen was extended with Beltsville Thawing Solution and contained 3 × 109 sperm/dose. A total of 73 multiparous sows were inseminated twice. Fertility was measured by conception rate, farrowing rate, litter size and number of piglets born alive following AI. Sperm quality and fertility data were analysed using one-way ANOVA. Spearman’s rank correlation was used to determine the relationship between sperm quality and fertility traits. The mean values for total sperm motility ranged from 93.5 to 96.8%. Progressive and rapid sperm motility differed significantly (P &lt; 0.05) among the boars. However, no significant differences were found for sperm velocity traits. The mean values for morphologically normal sperm ranged from 47.8 to 60.9% and live sperm ranged from 71.8 to 77.2%, but did not differ significantly among the boars (P &gt; 0.05). Conception rate from different boars varied (P &lt; 0.05) from 63.6 to 93.3%. Of all fertility traits studied, conception rate was significantly related to total sperm motility rate (r = 0.34, P &lt; 0.0029), progressive motility (r = 0.29, P &lt; 0.0141) and rapid motility (r = 0.34, P &lt; 0.0032), although relatively low. There was a low positive relationship between morphologically normal sperm and fertility traits (P &gt; 0.05). In conclusion, total, progressive, and rapid sperm motility rate were the only sperm traits significantly related to conception rate. Conversely, litter size and number born alive were not correlated with sperm motility, viability, or morphology traits.