<div>Background: The beta-defensin family antigens present on sperm surface suggestively play pivotal role in sperm fertility by aiding in various steps of sperm maturation, motility, capacitation, immune recognition and sperm-oocyte interaction in female tract. It is imperative to explore genetic polymorphisms to build a better understanding of individual variation in male fertility. The experiment was designed to identify DNA polymorphism in Sperm Associated Antigen 11B (SPAG11B) gene and to analyze association between genetic variants with sperm quality traits in buffalo bulls (Murrah) in ICAR-National Dairy Research Institute, Karnal.&nbsp; 
 
Methods: Genomic DNA was extracted from hundred and thirty Murrah bulls. A 395 base pair region covering partial Intron 2, Exon 3 and Partial Intron 3 of bovine SPAG11 gene was amplified and genotyped using PCR-RFLP method. The PCR amplified product was purified, sequenced and further ClustalW analysis was done to align edited sequence with reported Bos taurus sequence (AC_000158.1). Gene and genotype frequencies, effective number of alleles, level of heterozygosity and polymorphic information content of various genotypes were estimated by standard procedure. Seminal parameters (Post thaw sperm motility, sperm viability, acrosomal integrity, HOST and abnormality) were estimated and statistical analysis was carried out.&nbsp; 
 
Result: A novel SNP (G&gt;A substitution) at 2266 base of the SPAG11B gene was identified by sequencing. The fragment of SPAG11 gene containing 395 base pairs was amplified using PCR and digested with restriction enzyme i.e. MunI which showed three distinct genotypes viz., AA (266 bp and 107 bp fragment), AG (307 bp, 266 bp and 107 bp) and GG (373 bp fragment). Least squares means of seminal parameters for the SNP was estimated and compared after correction for non-genetic factors. Between genotypes, significant differences were found only for acrosomal integrity and was highest in AG (74.22&plusmn;0.72) followed by AA (72.6&plusmn;0.89) and GG (71.12&plusmn;0.97), respectively. The identified novel SNP of SPAG11 gene showed significant association with acrosome integrity with genotype AG being superior to other genotypes. However, an association cannot be established with other seminal parameters with this SNP, further studies are required in order to validate the impact of g.2266G&gt;A on sperm quality traits in a large population.</div>

SNP Identification in Sperm Associated Antigen 11B Gene and Its Association with Sperm Quality Traits in Murrah Bulls

Background: This study investigated the effects of Requimod (R848), a TLR7/8 activator on sperm quality, energy metabolism and the separation efficiency of X- and Y-chromosome-bearing sperm in frozen-thawed bull semen. 
&nbsp; 
Methods: Bull semen samples were divided into six groups: A control group and five experimental groups treated with different concentrations of R848 (0.05, 0.5, 1, 2 and 4 &mu;mol/L). Following incubation at 37oC for 60 min by upstream methods, sperm quality parameters were assessed for upper- and lower-layer sperm. Further evaluations included acrosomal integrity, plasma membrane integrity, mitochondrial activity and ATP content. 
&nbsp; 
Result: The results demonstrated that treatment with 0.5 &mu;mol/L R848 significantly reduced sperm quality parameters in lower-layer X-sperm. This concentration also inhibited mitochondrial activity and ATP content in X-sperm. However, no significant differences in acrosomal or plasma membrane integrity were observed between the control and experimental groups. Additionally, the R848 treatment had no significant effects on sperm quality parameters, acrosomal integrity, or plasma membrane integrity of the upper-layer Y-sperm compared to the control group. Flow cytometric analysis of separated sperm indicated that 76.13% of the sperm in the upper layer were Y-sperm, while 61.78% of the sperm in the lower layer were X-sperm. These findings suggest that 0.5 &mu;mol/LR 848 effectively enhances the separation of X- and Y-sperm in frozen-thawed bull semen.

Effects of TLR7/8 Activator on Sperm Quality, Energy Metabolism and the Separation Efficiency of X- and Y-Chromosome-bearing Sperm in Frozen-thawed Bull Semen

<div style="text-align: justify;">This study aimed to investigate the influence of negative pressure on boar semen quality during liquid preservation at 17&deg;C. Semen samples from ten large white boars were collected and pooled, divided into four equal parts, and diluted with Modena containing 0.4% (w/v) of bovine serum albumin. The semen samples were placed in a closed container with valve, and a negative pressure was applied for 2&ndash;5 minutes using a vacuum pump with a barometer. The control group had no treatment, the P200 group was treated at 200 mbar, the P400 group at 400 mbar, and the P800 group at 800 mbar. During liquid preservation, sperm motility, total antioxidant capacity, and semen H2O2 content were analyzed every 24 h. The effective survival time of boar semen during preservation was evaluated. The results indicated that a suitable negative pressure decreased the effects on reactive oxygen species on boar sperm quality during liquid preservation compared with the control group. Among all the groups, the 400 mbar negative pressure group had the highest sperm motility, total antioxidant capacity, and the percentage of spermatozoa with high mitochondrial membrane potential. The P400 group also had semen H2O2 content than the other groups. A suitable negative pressure improves sperm quality by reducing oxidative stress and the respiratory metabolism of sperm, and the optimum negative pressure is 400 mbar.</div>

Effects of negative pressure on boar semen quality during liquid storage at 17&deg;C

The objective of this study was to investigate the effect of using different extenders viz. Androhep, Safecell Plus and SUS during cryopreservation on quality of frozen-thawed boar semen. Semen volume, sperm motility, sperm concentration, percentage of morphologically abnormal spermatozoa, total number of spermatozoa per ejaculate and activity of the enzyme aspartate aminotransferase (AST) were assessed in fresh semen collected from 39 fertile AI boars. Semen from each boar was divided into three portions and diluted 1:1.5 in extender Androhep, Safecell Plus and SUS and keep at 17&deg;C for 15-h holding time before cryopreservation. Then sperm was cryopreserved. Straws were thawed in a water bath at 38&deg;C for 40s and post-thaw sperm motility with AST activity was assessed. &nbsp;Significant difference in post-thaw sperm motility was found between extender Androhep and Safecell Plus (P&lt;0.05). AST activity did not differ significantly between tested extenders (P&gt;0.05). In conclusion, the results of the study indicate that using Safecell Plus extender during holding period before cryopreservation significantly affected post-thaw sperm motility.

Effect of different extenders on quality of frozen-thawed boar semen

<div style="text-align: justify;">The present study was aimed to investigate the effects of negative pressure applied before storage on the quality and fertilization ability of boar semen. Boar semen samples were collected and pooled, and diluted with Modena solution containing 0.4% (w/v) of bovine serum albumin. Negative pressure was applied for 2&ndash;5 min using a vacuum pump with a barometer. The pressure applied were 0 (Control), -0.02 MPa (P200), -0.04 MPa (P400), and -0.08 MPa (P800). The sperm motility, acrosome integrity and sperm fertilizing ability were evaluated. Application of &ndash;0.04 MPa improved the sperm motility, acrosome integrity and fertilizing ability, compared with the other groups. The sperm motility and acrosome integrity decreased with increasing storage time in vitro. After 5 days, the sperm motility and acrosome integrity of the P400 group were all higher than those of the other groups (P &lt; 0.05). The cleavage rate (64.5% &plusmn; 2.4%) and blastocyst development rate (33.9% &plusmn; 2.8%) for semen stored for 7 days were similar to those of fresh semen. In conclusion, application of &ndash;0.04 MPa before liquid storage at 17&deg;C can improve the quality and fertilization ability of boar semen.</div>

<div style="text-align: justify;">Effects of negative pressure applied before storage (liquid storage, 17&deg;C) on boar semen quality and fertilization ability</div>

<div>Background: Fertility traits in pigs made a restricted progress through traditional selection. Applying marker assisted selection could improve these traits. The aim of the study was to investigate the effects of candidate genes Estrogen Receptor (ESR), Follicle Stimulating Hormone Beta (FSHB) and Prolactin Receptor (PRLR) on sperm quality traits of Landrace and Yorkshire boars under tropical conditions in Northern Vietnam. 
 
Methods: A total of 6306 ejaculates from 140 boars (92 Landrace and 48 Yorkshire) were collected to estimate sperm ejaculate volume (VOL), spermatozoon motility (MO), sperm concentration (CO) and total number of spermatozoon in ejaculate (NT). Ear tissue samples were collected for genotype identification of SNP present in ESR, FSHB and PRLR genes using PCR-RFLP technique. A mixed model was used to test the effect of different genetic and non-genetic factors on the sperm quality traits. 
 
Result: The genotype frequencies of ESR, FSHB and PRLR were in Hardy-Weinberg equilibrium for each breed. Age of boars and month of the year had significant effect on all the sperm quality traits (P&lt;0.01) while effect of breed was found to be non-significant on all the sperm quality trait. FSHB gene significantly (P&lt;0.05) affected VOL, MO and CO. Boars with BB genotype showed positive effect on VOL but negative effect on MO and CO in comparison with AA genotype boars. ESR gene showed only effect on VOL while PRLR affected only MO. Boars with AA and AB genotypes of ESR gene had a significantly higher VOL than those with BB genotype (P&lt;0.05). For PRLR gen, AB genotype was associated with higher MO than AA genotype (P&lt;0.05). These results suggest a possibility of using the existing polymorphisms in ESR, FSHB and PRLR genes to improve some sperm traits of Landrace and Yorkshire boars.</div>

<div>Effect of ESR, FSHB and PRLR Genes on Sperm Traits of Landrace and Yorkshire Boars in the Tropical Environmental Conditions of Vietnam</div>

Background: Aloe vera is a perennial herbaceous plant that resembles like a cactus that thrives in arid, warm climates. It has been known and used for centuries for its health, beauty, medicinal and skin care properties. Though it is used for treatment of variety of ailments and improves the body&rsquo;s physiology but continued treatment of fresh extract Aloe vera lowered sperm concentration, motility andpercentage viability, resulting in higher sperm abnormalities. The study was carried out to investigate the effect of different percentage of dried Aloe vera leaf powder on sperm quality of crossbred boar (75% Yorkshire x 25% Zovawk) with GEPS extender. 
&nbsp; 
Methods: The study was conducted on three mature, Large White Yorkshire (LWY) and Zovawk (75% LWY and 25% Zovawk) crossbred boars. A total of 45 ejaculates, 15 from each boar were taken for the study. Each ejaculate was combined 1:1 with extender (GEPS) and divided into three aliquots, each of which was diluted with 0%, 5% and10% Aloe vera powder (PH 7.4) and stored for 48 hours at 17&deg;C in a BOD incubator. At 0, 6, 12, 24 and 48 hours after preservation, the stored semen sample was tested for progressive motile sperm by traditional method, live sperm by Eosin-Nigrosin staining, acrosome integrity by Giemsa staining andsperm membrane integrity by HOST test, total protein, sperm membrane protein, cholesterol, ALT and AST level. 
&nbsp; 
Result: The per cent sperm motility, live sperm, acrosomal integrity and plasma membrane integrity of boar spermatozoa was decreased while preserving the semen sample by using GEPS extenders along with 5% and 10% of dried Aloe vera leaf powder at 17&deg;C for 48 hours. The level of total protein, ALT and AST in seminal plasma was increased while preserving the semen sample by using GEPS extenders along with 5% and 10% of dried Aloe vera leaf powder at 17&deg;C for 48 hours. The level of sperm membrane protein and cholesterol of boar spermatozoa decreased while preserving the semen sample by using GEPS extenders along with 5% and 10% of dried Aloe vera leaf powder at 17&deg;C for 48 hours. The total per cent of sperm abnormalities was increased while preserving the semen sample by using GEPS extenders along with 5% and 10% of dried Aloe vera leaf powder at 17&deg;C for 48 hours. In conclusion, the dried Aloe vera leaf powder @ 5% and 10% level having detrimental effect on semen quality of crossbred (75% Large White Yorkshire and 25% Zovawk) boar while preserving with ZEPS extender at 17&deg;C. The quality of boar semen was deteriorated chronologically at 0, 6, 12, 24 and 48 hours preservation with GEPS extender along with 5% and 10% of dried Aloe vera leaf powder at 17&deg;C. 
&nbsp;

Effect of Different Percentage of Dried Aloe Vera Leaf Powder on Quality of Boar Semen

Background: Sperm quality plays an important role in the animal industry. The study of the sperm quality of Congjiang Xiang pigs is limited and the Rh123/PI double staining has not been used to detect the sperm quality of boars. Thus, the study aimed to evaluate the sperm quality of the Congjiang Xiang pigs and the feasibility of the flow cytometry (FCM) method in assessing sperm quality in pigs. 
 
Methods: Semen samples were evaluated for routine indicators of semen quality and using FCM with single staining and double staining to analyze the integrity of membrane and acrosome, the mitochondrial function. The sperm quality of the Congjiang Xiang pigs was compared with the Large White pigs. 
 
Result: The semen of the Congjiang Xiang pigs had a lower sperm concentration than that of the Large White pigs (P&lt;0.01). There were no significant differences in sperm vitality, sperm motility and abnormality between the two breeds (P&gt;0.05). Single sperm staining revealed a lower sperm acrosomal integrity in the semen of the Congjiang Xiang pigs than that of the Large White pigs (P&lt;0.01). No significant difference between the two breeds in membrane integrity and mitochondrial function was observed (P&gt;0.05). Rh123/PI double staining demonstrated that the rate of live sperm with a normal mitochondrial function of the Congjiang Xiang pigs was significantly higher than that of the Large White pigs (P&lt;0.05). The rate of necrotic sperms in the semen of the Congjiang Xiang pigs was significantly lower than that in the semen of the Large White pigs (P&lt;0.05). Correlation analysis revealed a positive association between the rates of live sperm in the semen of the congjiang xiang pigs and their sperm motility (P=0.051). Our results suggest that Rh123/PI double staining with flow cytometry can provide more reliable information to assess the boar sperm quality.

Evaluation of the Congjiang Xiang Pig&rsquo;s Sperm Quality by Flow Cytometry

Background: Moringa leaf extract (MLE) is one of the natural ingredients that can improve the quality of preserved semen by preventing sperm lipid oxidation and increase the sperm storage time. The aim of the present study is to evaluate the effect of Moringa oleifera leaves extract on boar semen quality during short-term preservation at liquid state. 
&nbsp; 
Methods: Four mature healthy Large White Yorkshire boars of about 1.5-2 years age group having good reproductive health was selected for the study. Ten ejaculates were collected from each boar. Each ejaculate was divided into 4 aliquots (equal volume and sperm concentration) and Moringa oleifera leaves extract&nbsp;(MLE) @ 0 &micro;g/ml (control), 100 &micro;g/ml, 200 &micro;g/ml and 300 &micro;g/ml w/v was mixed in each aliquot after extension of semen samples with beltsville thaw solution (BTS) extender. The semen was evaluated for initial motility, percent live sperm, sperm abnormalities, sperm membrane integrity, acrosomal integrity and level of lipid peroxidation. Ten gilts/sows were artificially inseminated in each group with the liquid preserved semen to record the conception rates and litter size. 
&nbsp; 
Result: The percent sperm motility, live sperm, plasma membrane integrity and acrosomal integrity of boar spermatozoa were decreased significantly (P&lt;0.01) while preserving the semen sample by using BTS extenders along with 100 &micro;g/ml,200 &micro;g/ml and 300 &micro;g/ml Moringa oleifera leaves extract at 18&deg;C for 48 hours. The mean percent of motile sperm, live sperm, intact plasma membrane, intact acrosome among the groups i.e., 100 &micro;g/ml, 200 &micro;g/ml and 300 &micro;g/ml Moringa oleifera leaves extract were increased significantly (P&lt;0.01) as compared to control at 24 and 48 hours of preservation. The mean percentage of sperm abnormalities and lipid peroxidation among the groups i.e., 100 &micro;g/ml, 200 &micro;g/ml and 300 &micro;g/ml Moringa oleifera leaves extract were increased significantly (P&lt;0.05) at 0, 24 and 48 (P&lt;0.01) at hours of preservation as compared to control group. The supplementation of Moringa oleifera leaves extract in BTS extender at concentration of 100 &micro;g/ml, 200 &micro;g/ml and 300 &micro;g/ml significantly improved the Large White Yorkshire boar&rsquo;s semen quality during short term preservation at 18&deg;C as compared to control. The conception rate and average litter size at birth among the groups were increased significantly (P&lt;0.01) as compared to control (C). The significantly (P&lt;0.01) higher conception rate and litter size at birth was recorded in T3 group i.e. 300 &micro;g/ml Moringa oleifera leaves extract supplemented semen. In conclusion, the concentration of 300 &micro;g/ml Moringa oleifera leaves extract was found to be better among other concentrations. 
&nbsp;

Effect of Moringa oleifera Leaves Extract on Quality of Preserved Boar Semen

<div>Background: Artificial Insemination in pigs is much easier than other domestic species but the training of boar for the collection of semen is a big constraint. Certain boars are not acquainted with the fixed steel dummy sow and the surrounding environment. Hence, the dummy sows brought in the boar&rsquo;s pen. The steel dummy sow in comparison to wooden is too heavy and found difficult to bring in a boar&rsquo;s pen. Moreover, the wooden dummy sows are locally made and it becomes cost-effective too. The present study was designed to assess the libido and semen quality of boars by using locally made low-cost portable wooden dummy sows. 
 
Methods: The study was conducted on 16 numbers of Large White Yorkshire (LWY) and Mizoram local Zovawk crossbred boars at the age of 10-12 months. The semen collection was done by using a homemade portable &ldquo;wooden dummy sow&rdquo; and steel made &ldquo;fixed dummy sow.&rdquo; The training was done every morning before feeding and semen was collected twice a week by using the gloved hand method when the boars mounted over the dummy sow by holding protruded and erected corkscrew end of the penis firmly. During collection, libido characteristics of the boars were recorded and semen quality was determined.&nbsp;&nbsp; 
 
Result: The duration of training taken by the boars for semen collection was significantly (P&lt;0.01) less in portable wooden dummies. The sexual behaviour and other behavioural traits were better when boars were trained on a portable wooden dummy. The mean semen volume, sperm mass activity, pH and sperm concentration of LWY crossbred boar was not significantly&nbsp;different between fixed steel and portable wooden dummy. The percentage of initial sperm motility and intact acrosome of boar spermatozoa was significantly (P&lt;0.05) higher in portable wooden dummy. In&nbsp; conclusion, the homemade portable &ldquo;wooden dummy sow&rdquo; was found to be very effective for the training and collection of semen from breedingboars.</div>

SNP Identification in Sperm Associated Antigen 11B Gene and Its Association with Sperm Quality Traits in Murrah Bulls

Effects of TLR7/8 Activator on Sperm Quality, Energy Metabolism and the Separation Efficiency of X- and Y-Chromosome-bearing Sperm in Frozen-thawed Bull Semen

Effects of negative pressure on boar semen quality during liquid storage at 17°C

Effect of different extenders on quality of frozen-thawed boar semen

Effects of negative pressure applied before storage (liquid storage, 17°C) on boar semen quality and fertilization ability

Effect of ESR, FSHB and PRLR Genes on Sperm Traits of Landrace and Yorkshire Boars in the Tropical Environmental Conditions of Vietnam

Effect of Different Percentage of Dried Aloe Vera Leaf Powder on Quality of Boar Semen

Evaluation of the Congjiang Xiang Pig’s Sperm Quality by Flow Cytometry

Effect of Moringa oleifera Leaves Extract on Quality of Preserved Boar Semen

Assessment of Libido and Semen Quality of Boar by using Low-cost Portable Wooden Dummy Sow