<div style="text-align: justify;">Study was conducted to compare the semen quality and fertility of liquid stored semen for three days and frozen-thawed semen in the north-eastern region of India. For liquid semen, the semen ejaculates were extended in Beltsville Thawing Solution (BTS) extender and preserved at 17&deg;C for three days. For cryopreservation, semen was diluted Lactose-egg yolk-glycerol extender and frozen in straw using programmable freezer with freezing rate of 40&deg;C/min from -6 to -140&deg;C. The preserved evaluated for sperm motility, viability, plasma membrane integrity and fertility. The results revealed that the liquid stored semen has maintained the sperm motility and viability up to day 3 without significant reduction. Similarly the plasma membrane integrity did not differ significantly up to day 2, but it was significantly (P&lt;0.05) reduced on days 3 in liquid stored semen. After freezing and thawing, the mean sperm motility, viability and plasma membrane integrity were 58.25 &plusmn; 2.96%, 64.75 &plusmn; 2.47% and 47.06 &plusmn; 2.02%, respectively. These parameters were significantly (P&lt;0.01) lower as compared to the liquid stored semen from day 0 to day 3. After insemination with liquid semen, the farrowing rate was 77.7%, 80.76%, 73.07% and 69.8%, respectively from day 0, day1, day 2 and day 3. The pregnancy rate, farrowing rate and litter size did not differ significantly among different days of liquid storage. These parameters were significantly (P&lt;0.01) lower in frozen semen as compare to that of liquid stored semen. The study concluded that the liquid semen stored up to three days is more efficient than frozen-thawed semen in terms of preserving sperm quality and fertility.</div>

<div style="text-align: justify;">Semen quality and fertility of liquid stored and frozen-thawed semen in crossbred pigs of North-Eastern India</div>

<p style="text-align: justify;">The present experiment was conducted to evaluate the effect of two different beats {100 and 150 BPM (Beat Per Minute)} of musical sound signals {intensity below 85 dB(decibel)} on reproductive performance of Hariana bulls maintained at semen biology lab of DDD (District Dairy Demonstration) Farm within the premises of Instructional Livestock Farm Complex (ILFC). The present experiment was carried out for an aggregate time period of three and half months (within autumn season) and was accomplished in three phases. Same four Hariana bulls were used as experimental animals in three different phases of experiments to avoid individual effect of bulls as well as to overcome the limitations of limited numbers of bulls. During the first (control) phase of experiment the bulls were not exposed to any additional source of sound (T1) except the normal environmental sound in the semen collection area. In second and third phase, bulls were exposed to a musical instrumental sound signal of 100 (T2) and 150 (T3) BPM with intensity below 85 dB. &nbsp;Each phase of the experiment was done for a period of one month (four weeks) one after another in continuation but the third phase was started after a gap of three weeks from the second in order to cancel out the persistent effect of sound exposure to bulls during second phase of investigation. The effect of exposure of sound signals on volume (6.58&plusmn;0.29) ml, concentration (1018.72&plusmn;49.92) millions/ml, mass (3.98&plusmn;0.07) and progressive motility (72.97&plusmn;0.80%), head (1.93&plusmn;0.21%), middle piece (0.63&plusmn;0.15%), and tail (2.56&plusmn;0.28%) of sperm and HOST (79.22&plusmn;0.69) was highly significantly better (P&lt;0.01), with an exposure of a sound signal of 150 BPM with intensity below 85 dB.&nbsp;</p>


<p style="text-align: justify;">Effect of exposure of sound signals in semen collection area&nbsp;on quantity and quality of semen in Hariana bulls</p>


<div style="text-align: justify;">In cattle, sex selection has a significant economic impact when it improves herd capacity of milk or meat production and is commonly used for the production of calves of the desired gender. &nbsp;New separation techniques which present both better accuracy and low costs are necessary. Density gradient centrifugation might be an approach to sexing spermatozoa because of the additional DNA content and volume of X-bearing sperm head. The present study was designed to apply the modified percoll discontinuous density gradient for X-sperm enrichment in cross bred bovine semen. We attempted to amplify the level of X-sperm enrichment by using double centrifugation in percoll gradient. In addition, effect of percoll density gradient on fertility of semen was also evaluated at three stages; before centrifugation (stage I), after 1st centrifugation (stage II) and after 2nd centrifugation (stage III) respectively and ultrasonography was carried out to confirm the sex of fetus at 60-90 days post AI. Semen was collected from two cross bred bovine bulls and 34 cows were inseminated with liquid X-sperm enriched semen after double centrifugation in percoll gradient. 22 cows were inseminated with non-sexed fresh liquid semen (control). Results showed that the volume, progressive motility, live sperms, concentration and HOST reactive sperms significantly (P&lt;0.05) decreased at stage II and stage III compared to stage I. However, no significant (P&gt;0.05) change was observed in pH and mass motility of semen at all three stages. There was non-significant effect on per cent spermatozoa with intact acrosomal membrane and fully damaged acrosomal membrane at all three stages however, partially damaged acrosomal membrane sperms were significantly (P&lt;0.05) decreased at Stage II and Stage III compared to Stage I. The conception rate significantly (P&lt;0.05) decreased with X-enriched semen (41.17%) compared to non-sexed liquid semen (59.09%). The female sex ratio of calves significantly (P&lt;0.05) increased to 66.66% with X- sperm enriched semen compared to the male percentage of 33.33% with sexed semen and female percentage of 46.15% with non-sexed semen.</div>

<div style="text-align: justify;">X-sperm Enrichment of Bovine Semen by Percoll Density Gradient Method and Its Effect on Semen Quality, Sex Ratio and Conception Rate</div>

<div><strong>Background: </strong>Zinc is found in very high concentration in semen and male reproductive tract and acts as a hormone receptor modulator for testosterone production in Leydig cells. Thus, it plays a very important role in male reproduction. Moreover, Indian soils are very deficient in zinc as per earlier reports and recent studies have shown that supplementation of zinc has improved qualitative as well as the quantitative aspects of semen quality in bulls. So, a study has been planned to study the effect of organic zinc supplementation on semen production and quality in kankrej bulls.<br />
<br />
<strong>Methods:</strong> A total of 8 healthy Kankrej bulls under semen collection were selected for the study. Basal diet (concentrate mixture, green and dry fodder) was fed during pre-supplementation and zinc propionate @ 100 ppm/bull was supplemented to the basal diet during post-supplementation for a period of 60 days to complete one spermatogenic cycle. A total of 128 ejaculates (64 pre-supplementation and 64 post-supplementation) were collected.&nbsp;<br />
<br />
<strong>Result: </strong>Mean semen volume was non-significant (P&gt;0.05) between the treatment groups. Mean sperm concentration was 1325.68&plusmn;80.08 and 2162.36&plusmn;112.0 million/ml pre- and post-supplementation, respectively. Feeding of zinc propionate significantly (P&lt;0.01) increased semen sperm concentration. The average mass motility of sperms was significantly (P&lt;0.01) higher during post-supplementation (3.92%) than the pre-supplementation (3.78%). The overall live sperms were significantly (P&lt;0.01) higher during post-supplementation (73.33&plusmn;0.91%) than the pre-supplementation (63.53&plusmn;1.03%). It may be concluded that supplementation of 100 ppm of zinc propionate in the diet of Kankrej bulls improved semen quality in terms of quantitative and qualitative characteristics of semen.</div>


<p>Effect of Organic Zinc Supplementation on Semen Production and Quality in Kankrej Bulls</p>


<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

<div><strong>Background:</strong> Quality determination of frozen semen before using for Artificial Insemination (AI) in cattle is of paramount importance. The success of cattle AI programs is largely depends on the use of good quality semen and thus assessment of semen quality parameters before AI will improve the conception rate and reduces the loss associated with repeated breeding. Among the various semen quality indicators, sperm motility is one of the important parameter and it should be evaluated immediately after semen collection (before freezing) and 24 hrs after freezing as per minimum standard protocol in frozen semen production. Though the motility is a major determinant of spermatozoa ability to reach the site of fertilization and it is affected by cryopreservation and subsequent poor cold storage facilities, motility is not frequently evaluated under field conditions. In the current scenario, many of our government AI sub-stations in remote villages are not equipped with microscope facility for semen quality evaluation before AI.&nbsp;<br />
<br />
<strong>Methods:</strong> In this study, we used foldscope, an origami-based paper microscope which give optical quality similar to conventional microscopes (magnification of 140X and 2-micron resolution), to evaluate the frozen semen motility at field level. In this study, the motility of spermatozoa was observed under foldscope and concluded that foldscope is very useful to asses the motility befor AI and improve the fertility in dairy animals.&nbsp;<br />
<br />
<strong>Result: </strong>We revealed that, utility of foldscope to assess the motility of spermatozoa was evaluated under field conditions and observed that 46% of sperms were motile. Further, evalaution of sperm motility before AI is not very common practice due to lack of microscope in many AI centres. Therefore, foldscope will be very useful at field conditions to avoid AI using poor quality semen and subsequent loss associated with repeat breeding problems.</div>


<p>Assessment of Frozen Semen Quality through Foldscope Microscopy- A Novel Application of Frugal Science to Reduce the Infertility Rate</p>


<p>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.</p>


<p>Effect of different extenders on quality of frozen-thawed boar semen</p>


<div>An investigation was conducted to detect single nucleotide polymorphism in Inhibin alpha gene (INHA) and to analyse the association between the observed polymorphisms with semen quality traits in Murrah buffalo bulls. Genomic DNA was isolated from blood samples of 103 bulls, the amplified PCR products were further subjected to polyacrylamide gel electrophoresis. Band patterns were characterised by the number of bands, mobility shifts and scored manually. Representative PCR products giving unique SSCP patterns were custom sequenced using automated ABI DNA Sequencer to confirm the mobility shift in each pattern. Sequence data were analysed using DNA Baser and Bio edit software Clustal omega multiple alignments for detecting single nucleotide polymorphisms in comparison to observed sequence with the Bubalus sbubalis INHA gene reference sequence. INHA gene, exon1 was monomorphic, while exon 2 was found to be polymorphic. For simple interpretation of SSCP patterns, INHA exon 2 was divided into two fragments. The first fragment of INHA exon 2 (INHA exon 2 FR1) revealed a transition at the C267119T position and a transversion at the G267033T position in the coding region in comparison to Bubalus bubalis reference sequence. The second fragment of INHA exon 2 (INHA exon 2 FR2) exhibited one transition at T266510C position in comparison to the Bubalus bubalis reference sequence. The SSCP variants of the first fragment of INHA exon2 had a significant effect on semen volume per ejaculate (P&lt;0.05) and sperm concentration (P&lt;0.01). The SSCP variants of the second fragment of INHA exon 2 had a significant effect on sperm concentration (P&lt;0.01) and functional membrane integrity (P&lt;0.05). The observed association between SSCP variants in INHA gene with semen volume per ejaculate, sperm concentration and functional membrane integrity suggests the possibility of using these genetic variants in INHA gene as prognostic markers for selecting Murrah bulls with high semen quality traits.</div>


<div>Polymorphism in inhibin alpha gene and its association with semen quality traits in Murrah bulls</div>


<div style="text-align: justify;">Bull fertility is an important factor for genetic improvement; therefore prediction of fertility an early age of the bulls by using genetic markers is a goal for livestock breeding. Zinc finger protein280BY linked gene is predominantly expressed in testis and has a major role in spermatogenesis. An investigation was carried out to detect the genetic variants in the male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffaloes. Polymerase Chain Reaction-Single Strand Conformation Polymorphism technique (PCR-SSCP) and DNA sequencing methods revealed three different SSCP band patterns (AA, BB, CC); one transition (C10375T) &nbsp;and an indel (insertion of T at 10460-10461). The associations of the genetic variants with semen volume, sperm concentration, Hypo Osmotic Swelling Test (HOST) in fresh and frozen semen were observed. AA and CC genotype bulls had high volume (P&pound;0.05) of semen per ejaculation than BB genotype bulls. BB genotype bulls had high sperm concentration (106/mL) per ejaculate in comparison to AA and CC genotype bulls (P&pound;0.05). AA and BB genotype bulls had a higher per cent of HOST (P&pound;0.05) reacted sperms of fresh semen and per cent acrosomal integrity of frozen semen in comparison to CC genotype bulls. The observed association between SSCP variants in MSY region of ZNF280BY gene with semen quality parameters indicated the possibilities of using ZNF280BY as a candidate gene for identification of markers for semen quality traits in Murrah buffaloes.</div>

<div style="text-align: justify;">Genetic variants in male specific region (MSY) of <em>ZNF280BY</em> gene and their association with semen quality traits in Murrah buffalo bulls</div>

<p>Semen quality and fertility of Bama miniature pig and the effect of season on these parameters were determined in this study. Results were as follows: the overall mean ejaculate volume, 55.71 &plusmn; 15.21 ml; sperm concentration, 0.66 &plusmn; 0.22 &times; 108/ml; sperm motility, 65.39% &plusmn; 16.45%; abnormal sperm percentage, 12.16% &plusmn; 7.70%; farrowing rate, 76.44%; litter size, 7.74 &plusmn; 2.46. It showed that the semen quality and fertility of Bama miniature pigs were lower than the normal level of the larger commercial and local breeds. In terms of the effect of season, we noted that the semen quality and fertility of Bama miniature pigs were not worst in summer. This may indicate that Bama miniature pig is a relative heat-tolerant&nbsp;breed in comparison to larger commercial and local breeds. These findings in this study could provide reference material for improve management and husbandry in practice and for using Bama miniature pigs as animal model especially in male fertility studies.</p>


Semen quality and fertility of liquid stored and frozen-thawed semen in crossbred pigs of North-Eastern India

Effect of exposure of sound signals in semen collection area on quantity and quality of semen in Hariana bulls

X-sperm Enrichment of Bovine Semen by Percoll Density Gradient Method and Its Effect on Semen Quality, Sex Ratio and Conception Rate

Effect of Organic Zinc Supplementation on Semen Production and Quality in Kankrej Bulls

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

Assessment of Frozen Semen Quality through Foldscope Microscopy- A Novel Application of Frugal Science to Reduce the Infertility Rate

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

Polymorphism in inhibin alpha gene and its association with semen quality traits in Murrah bulls

Genetic variants in male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffalo bulls

Semen quality and fertility of Bama miniature pig (Sus scrofa domestica)