Search results for: post-thaw semen quality

Authors: Abdelrasoul, Khalid Ahmed Elrabie

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The study was conducted on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to determine the effect of exercise on the sexual behavior and semen quality. Fourteen Sahiwal bulls were classified into two groups of seven each. Group-1, bulls were exercised by walking in a bull exerciser once a week one hour before semen collection, whereas bulls in group-2 were exercised daily. Sexual behavior and semen quality traits studied were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-2 showed significantly (p < 0.01) higher value in RT (sec), DMT (sec), TTTM (sec), ES, PS, ITS, LS, semen volume, semen color density and mass activity.

Keywords: exercise, Sahiwal bulls, semen quality, sexual behavior

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Authors: Pankaj K. Jha, Ajeet K. Jha, Pravin Mishra

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Goat raising is a popular livestock sub-commodity of mixed farming system in Nepal. Besides food and nutritional security, it has an important role in the economy of many peoples. Goat breeding through AI is commonly practiced worldwide. It is a very basic tool to speed up genetic improvement and increase productivity. For the goat genetic improvement program, the government of Nepal has imported some specialized exotic goat breeds and semen. Some progress has been made in the initiation of selective breeding within the local breeds and practice of AI with imported semen. Importance of AI in goats has drawn more attention among goat farmers. However, importing semen is not a permanent solution at national level; rather, it is more important to develop and establish its own frozen semen production technique. Semen quality and its relationship with fertility are said to be a major concern in animal production, hence accurate measurement of semen fertilizing potential is of great importance. The survivability of sperm cells depends on semen quality. Survivability of sperm cells is assessed through visual and microscopic evaluation of spermatozoal progressive motility and morphology. In Nepal, there is lack of scientific information on seminal attributes of buck semen, its dilution, cooling and freezing technique under management conditions of Nepal. Therefore, the objective of this review was to provide brief information about breeding system, semen production and artificial insemination in Nepalese goat.

Keywords: artificial insemination, goat, Nepal, semen

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Authors: Aftab Ali

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Cryopreservation of semen is one of the key factors for a successful breeding business along with other factors. To achieve high fertility in boar, one should know about spermatozoa response to different treatments proceeds during cryopreservation. The running project is highly focused on cryopreservation and its effects on sperm quality parameters in both boar and bull semen. Semen sample from A, B, C, and D, were subjected to different thawing conditions and were analyzed upon different treatments in the study. Parameters like sperm cell motility, viability, acrosome, DNA integrity, and phospholipase C zeta were detected by different established methods. Different techniques were used to assess different parameters. Motility was detected using computer assisted sperm analysis, phospholipase C zeta using luminometry while viability, acrosome integrity, and DNA integrity were analyzed using flow cytometry. Thawing conditions were noted to have an effect on sperm quality parameters with motility being the most critical parameter. The results further indicated that the most critical step during cryopreservation of boar semen is when sperm cells are subjected to freezing and thawing. The findings of the present study provide insight that; boar semen cryopreservation is still suboptimal in comparison to bull semen cryopreservation. Thus, there is a need to conduct more research to improve the fertilizing potential of cryopreserved boar semen.

Keywords: cryopreservation, computer assisted sperm, flow cytometry, luminometry

Procedia PDF Downloads 148

Authors: Khalid Ahmed Elrabie Abdelrasoul

Abstract:

The present study was carried out on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to assess the effect of cooling using mist cooling and fanning on Sahiwal bulls in the dry hot summer season. Fourteen Sahiwal bulls were divided into two groups of seven each. Sexual behavior and semen quality traits considered were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-1 was the control, whereas group-2 (treatment group) bulls were exposed to mist cooling and fanning (thrice a day 15 min each) in the dry hot summer season. Group-2 showed significantly (p < 0.01) higher value in DMT (sec), ES, PS, ITS, LS, semen volume (ml), semen color density, mass activity, initial motility, progressive motility and live sperm.

Keywords: mist cooling, Sahiwal bulls, semen quality, sexual behavior

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Authors: Ajeet K. Jha, Pankaj K. Jha, Pravin Mishra

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The study was conducted to evaluate the freezability of crossbred bulls’ semen at early age. Semen of three consecutive collections at 7 days interval from 12 crossbred bulls 17 was evaluated. The age at first collection was 15 to 20 months. Evaluation of semen was done soon after collection. Triladyl, Minitub, Germany was used as extender and was frozen using standard semen freezing protocol. Post-thaw sperm motility was evaluated. Morphology of paraformaldehyde fixed spermatozoa was evaluated under differential interference phase contrast microscopy and the viability of spermatozoa was evaluated by using stain SYBR-14 (1 mM/ml) and propidium iodide (2.41 mM/ml) under an epifluorescent microscopy. Scrotal circumference was correlated with all possible measures in all groups of crossbred bulls. Volume of semen, sperm concentration, total number of spermatozoa, initial sperm motility, post-thaw sperm motility, proportion of normal spermatozoa and proportion of live spermatozoa were compared among individual bull within and between two groups of crossbred bulls. A significant positive correlation was observed between scrotal circumference and volume of semen and between scrotal circumference and the total number of sperm production per ejaculate (r = 0.72, p < 0.04). Significant variation was observed in different semen parameters among individual bulls within the same group (p < 0.05) but no significant variation was found between two groups of crossbred bulls. Out of 12 bulls, semen freezability of 10 bulls was found satisfactory while semen of 2 bulls (Local × Friesian) was unsatisfactory. In conclusion, crossbred bulls aged 18 months having scrotal circumference > 30 cm produce freezable quality semen.

Keywords: Bangladesh, crossbred bull, scrotal circumference, semen freezability

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Authors: Gatot Ciptadi, Mudawamah, R. P. Putra, S. Wahjuningsih, A. M. Munazaroh

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The success rate of Artificial Insemination (AI) application, particularly in the field at the farmer level is highly dependent on the quality of the sperms one post thawing. The objective of this research was to determine the effect of freezing method (-1oC/ minute) using Mr. Frosty system with commercial diluents on the post-thawing quality of Boer goat semen. Method use is experimental design with the completely randomized design (CRD) with 4 treatments of commercial diluter percentage (v/v). Freezing semen was cryopreserved in 2 main final temperatures of –45 oC (Freezer) and –196 oC (liquid nitrogen). Result showed that different commercial diluter is influenced on viability motility and abnormalities of Boer semen. Pre-freezing qualities of viability, motilities and abnormalities was 88.67+4.16 %, 66.33 +1.53 % and 4.67+ 0.57 % respectively. Meanwhile, post-thawing qualities is considered as good as standard qualities at least more than 40 % (51.0+6.5%). The percentage of commercial diluents were influenced highly significant (P<0.01).The best diluents ration is 1:4 (v/v) for both final sperms stocked. However freezing sperm conserved in -196 oC is better than –45 oC (i.e. motility 39.3.94 % vs. 51.0 + 6.5 %). It was concluded that Mr. frosty system was considered as the feasible method for freezing semen in the reason for practical purposes.

Keywords: sperm quality, goat, viability, diluteR

Procedia PDF Downloads 259

Authors: M. M. Islam, S. Sharmin, M. Shah Newaz, N. S. Juyena, M. M. Rahman, P. K. Jha, F. Y. Bari

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The aim of the present study was to select the high quality ram by measuring the scrotal biometry which has an effect on semen parameters. Ten rams were selected in the present study. Eight ejaculates were collected from each ram using artificial vagina method. Scrotal circumference was measured before and after semen collection on weekly basis using the Scrotal tape. Bio-metries of scrotum (scrotal length and scrotal volume) were calculated. Semen was evaluated for macroscopic and microscopic characteristics. The average estimated scrotal circumference (cm) and scrotal volume (cm3) in 8 different age groups were 17.16±0.05 cm and 61.30±0.70 cm3, 17.17±0.62 cm and 63.67±4.49 cm3, 17.22±0.52 cm and 64.90±4.21 cm3, 17.72±0.37 cm and 67.10±4.20 cm3, 18.41±0.35cm and 69.52±4.12cm3, 18.45±0.36cm and 77.17±3.81 cm3, 18.55±0.41 cm and 78.72±4.90 cm3, 19.10±0.30 cm and 87.35±5.45 cm3 respectively. The body weight, scrotal circumference and scrotal volume increased with the progress of age (P < 0.05). Body weight of age group 381-410 days (13.62+1.48 kg) was significantly higher than group 169-200 days (10.17±0.05 kg) and 201-230 days (10.42±1.18 kg) (p < 0.05). Scrotal circumference (SC) of age group 381-410 days (19.10±0.30 cm) was significantly higher (p < 0.05) than other groups. In age group 381-410 days, scrotal volume (SCV) (87.35±5.45 cm3) was significantly higher than other first five groups (p < 0.05). Both scrotal circumference and scrotal volume development was positively correlated with the increasing of body weight (R2= 0.51). Semen volume increased accordingly with the increasing of ages, varied from 0.35±0.00 ml to 1.15+0.26 ml. Semen volume of age group 381-410 days (1.15±0.26 ml) was significantly higher than other age groups (p < 0.05) except age group 351-380 days (p > 0.05). Mass activity of different age groups varied from 2.75 (±0.35) to 4.25 (±0.29) ml in the scale of 1-5. Sperm concentration, progressive motility (%),progressively improved according to the increasing of ages, but significant changes in these parameters were seen when the animals reaches the age 291 days or more (p < 0.05). However, normal spermatozoa (%) improved significantly from the age of 261 days or more. Mass activity (mass) was positively correlated with sperm concentration (R2=0.568) and progressive motility (%) (R2=0.616). The relationships of semen volume with body weight and scrotal measurements and sperm concentration indicate that they are useful in evaluating rams for breeding soundness and genetic improvement for fertility in indigenous ram.

Keywords: breeding soundness, ram, semen quality, scrotal biometry

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Authors: Pankaj Kumar Jha, Saroj Sapkota, Dil Bahadur Gurung, Raju Kadel, Neena Amatya Gorkhali, Bhola Shankar Shrestha

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The study was conducted to evaluate the seminal attributes, effectiveness of cooling process and post-thawed semen quality of a Nepalese indigenous Khari buck. Thirty-two ejaculates, 16 from each buck were studied for seminal attributes of fresh semen: volume, color, mass activity, motility, viability, sperm concentration, and morphology. The pooled mean values for each seminal attributes were: volume 0.7±0.3 ml; colour 3.1±0.3 (milky white); mass activity 3.8±0.4 (rapid wave motion with formation of eddies at the end of waves to very rapid wave motion with distinct eddies formation); sperm motility 80.9±5.6%; sperm viability 94.6±2.0%; sperm concentration 2597.0±406.8x106/ml; abnormal acrosome, mid-piece and tail 10.7±1.8% and abnormal head 5±1.7%. For freezing semen, further 6 ejaculates from each buck were studied with Tris based egg yolk citrate extender. The pooled mean values of motility and viability of post diluted semen for 90 and 120 minutes each for cooling and glycerol equilibration were 73.8±4.8%, 88.1±2.6% and 69.2±6.0%, 85.0±1.7%, respectively. The pooled mean values of post thaw motility and viability with advancement of preservation time were: 0hour 49.0±4.6%, 81.2±1.9%; 2nd day 41±2.2%, 79±1%; 5th day 41±2.2%, 78.6±0.9% and 10th day 41±2.2%, 78.6±0.9%. We concluded from the above study that the seminal attributes and results of post-thaw semen quality were satisfactory and in accordance with other work in foreign countries, which indicated the feasibility of cryopreserving buck semen. For more validation, research with large number of bucks, different types of diluents and freezing trials by removing seminal plasma followed by pregnancy rate is recommended.

Keywords: cryopreservation, Nepalese indigenous Khari (Hill goat) buck, post-thaw semen quality, seminal attributes

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Authors: I. P. Ogbuewu, I. F. Etuk, V. U. Odoemelam, I. C. Okoli, M. U. Iloeje

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The effect of dietary ginger rhizome meal on libido and semen characteristics of post-pubertal rabbit bucks was investigated in an experiment that lasted for 12 weeks. Thirty-six post-pubertal bucks were randomly assigned to 4 dietary groups of 9 rabbits each in a completely randomized design. Four experimental diets were formulated to contain ginger rhizome meal at 0 g/kg feed (BT0), 5g/kg feed (BT5), 10 g/kg feed (BT10), and 15g/kg feed (BT15) were fed ad libitum to the experimental animals. Results revealed that semen colour changed from cream milky to milky. Data on semen pH and sperm concentration were similar (p>0.05) among the dietary groups. Semen volume for the bucks in BT0 (0.64 mL) and BT5 (0.60 mL) groups were significantly (p<0.05) higher than those in BT10 (0.44 mL) and BT15 (0.46 mL) groups. Total spermatozoa concentration value was significantly (p<0.05) higher in BT0 and BT5 groups than those in BT10 and BT15 groups. Sperm motility and percent live sperm declined (p<0.05) progressively among the treatment groups. Percent dead sperm were significantly (p<0.05) lower for bucks in BT0 group than in BT10 and BT15 groups. Reaction time had a dose-dependent increase; however, the observed difference was not significant (p>0.05). These results indicate that the inclusion of ginger rhizome meal at 5-15g per kg feed in ration for post-pubertal rabbit bucks could cause mild depressive effect on semen production and quality.

Keywords: rabbits, semen, libido, ginger

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Authors: E. A. Babiker, S. A. Makawi

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The influence of the season (autumn, winter, and summer) on semen production in Nubian and Saanen bucks was studied. Seven mature bucks (4 Nubian and 3 Saanen) were used in this study to prepare semen samples which were collected with an artificial vagina. The samples were extended in Tris-egg yolk-glycerol-glucose extender, frozen, and stored in liquid nitrogen at –196 0C for 48 hours. Straws were thawed in water at –37 0C for 15 seconds before sperm evaluation (post-thaw sperm motility). There was a significant seasonal variation in both semen quantity (volume, concentration, and the total number of spermatozoa per ejaculate) and quality (percentage of sperm motility, percentage of post-thaw sperm motility, and dead spermatozoa). Greater ejaculate volumes were observed during summer and autumn in comparison to winter. Higher values of sperms concentration were observed during autumn, while the lowest sperm concentration values were observed during summer. Higher values of sperm motility were observed during autumn in comparison to summer. Lower values of dead spermatozoa were recorded during autumn, while the highest percentages of dead spermatozoa were observed during summer for the two breeds of bucks. The influence of season on post-thaw sperm motility was significant. Semen frozen during autumn and winter had the highest values, while during summer, lower mean values were observed. The best semen was produced during autumn and winter, while during summer, poor semen quality was recorded.

Keywords: season, Nubian, Saanen, semen production, Sudan

Procedia PDF Downloads 112

Authors: A. M. Raseona, D. M. Barry, T. L. Nedambale

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Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technology. This study evaluated the effectiveness of different extenders to preserve Nguni bull semen stored at controlled room temperature 24 °C for three days, as an alternative to frozen-thawed semen straws used for artificial insemination. Semen samples were collected from two Nguni bulls using an electro-ejaculator and transported to the laboratory for evaluation. Pooled semen was aliquot into three extenders Triladyl, Ham’s F10 and M199 at a dilution ratio of 1:4 then stored at controlled room temperature 24 °C. Sperm motility was analysed after 0, 24, 48 and 72 hours. Morphology and viability were analysed after 72 hours. The study was replicated four times and data was analysed by analysis of variance (ANOVA). Triladyl showed higher viability percentage and consistent total motility for three days. Ham’s F10 showed higher progressive motility compared to the other extenders. There was no significant difference in viability between Ham’s F10 and M199. No significant difference was also observed in total abnormality between the two Nguni bulls. In conclusion, Nguni semen can be preserved in Triladyl or Ham’s F10 and M199 culture media stored at 24 °C and stay alive for three days. Triladyl proved to be the best extender showing high viability and consistency in total motility as compared to Ham’s F10 and M199.

Keywords: bull semen, artificial insemination, Triladyl, Ham’s F10, M199, viability

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Authors: K. A. El-Battawy, W. S. El-Nattat

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The aim of the present investigation was to evaluate the impact of methionine on the preservation, acrosomal integrity, DNA integrity and post thawing motility of extended goat semen. Semen samples were diluted with a Tris-based extender containing the additive methionine 1.5, 2.5 and 5mM then the diluted samples were kept in glass tubes and cooled from 37°C to 5°C in a cold cabinet, and maintained at 5°C. Sperm motility (SM%), alive sperm (AS%), sperm abnormalities (SA%) acrosomal integrity and DNA integrity were determined at 5°C for periods of 0,24, 48and 72 h of liquid storage. Furthermore, the influence of methionine on post-thawing motility was assessed. The results elaborated that the addition of methionine and L-tyrosine particularly 2.5mM of methionine significantly improved SM% and reduced dead sperm %. Furthermore, the addition of 2.5mM methionine improved post-thawing motility (43.75 ± 1.25% vs. 32.50 ± 3.23 in the control group). Moreover, the frequency of acrosomal defects was lower in treated groups than in control. In conclusion, the addition of methionine induced remarkable physiological effects on goat semen quality during conservation for 7-days-long period at 5°C and improved its freezability.

Keywords: methionine, acrosome, semen, cryopreservation

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Authors: Abdurzag Kerban

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Preservation of semen had a major impact on sheep genetic breeding. The aim of this study was to evaluate the viability of ram spermatozoa after freezing pellet using cold surfaces made from cattle fat and paraffin wax. A pool of three to four ejaculates were pooled from six rams within a period of ten weeks. Semen was diluted in egg yolk-Tris diluent and processed in 0.25 ml straw and 0.1 ml pellets. Motility was evaluated after dilution, before freezing and post-thawing at 0, 1, 2 and 3 hour incubation. Viability index, acrosome integrity and leakage of intracellular enzymes (aspartat aminotransferase and alkline phosphatase) were also evaluated. Spermatozoa exhibited highly significant percentages of motility at 0, 1, 2 and 3 hours incubation after thawing and viability index in 0.25 ml straw and 0.1 ml pellets on cattle fat plate as compared to ram spermatozoa frozen on paraffin wax. In conclusion, cattle fat plate could be used as the cold surface of choice for freezing ram semen in form of pellets. Such form of frozen semen could be used as efficiently as semen frozen in straws. This simple method is economical with little expensive equipment or supplies, and may provide an efficient technique to cryopreserve ram spermatozoa in developing countries.

Keywords: ram semen, freezing, straw, pellet

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Authors: Jamshid Ghiasi Ghalehkandi, Naser Maheri Sis, Yahya Ebrahimnezhad, Shahin Hassanpour

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The objective of present study was to examine the effects of fresh garlic juice on semen malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx) and total antioxidant status (TAS) in male rats. Fifty-four male rats (230-250 g) were allocated into 3 treatment groups (each include 3 groups and 6 replicate). Group 1 served as water control. In group 2, rats were gavaged with 60 mg/kg garlic juice. In group 3, rats were offered 120 mg/kg garlic juice. Animals received treatments orally and ad libitum access to chow pellets and fresh water. After 4 weeks, animals were killed, testes were taken out and semen samples were used to determine MDA, SOD, GPx and TAS activity. According to the results, garlic juice (120 mg/kg) significantly declined semen MDA activity compared to control group (P<0.05). These results suggest that presumably garlic juice protects semen oxidation in rat testes.

Keywords: garlic juice, chromium chloride, semen, rat

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Authors: Muhammad Zubair, Maqbool Ahmad, Al-Hafizah Shafia Tehseen Gul, Shujait Ali

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The use of vitamins has significant effects on the reproductive system of mammals. The present study was conducted to investigate the useful effects of vitamin E on reproductive functions of Teddy bucks. For this purpose, 8 adult Teddy bucks were randomly divided into two treatment groups viz; A (control) and B (vitamin E with dose of 200 mg/kg BW/day). These treatments continued for 12 weeks. Semen quality parameters (volume, motility, sperm morphology and sperm DNA integrity) of experimental bucks of each group was evaluated on weekly basis, while testicular measurements (length, scrotal circumference and weights) were recorded at 0 and 12th week of experiment. Serum concentrations of male sex hormones (testosterone, LH, FSH) and cortisol were recorded fortnightly. Similarly, body weights of bucks were also measured fortnightly until completion of the study. The data were subjected to two-way analysis of variance, followed by Duncan test for multiple mean comparisons. Supplementation of vitamin E improved significantly (P<0.05) the semen quality parameter, body weights, testicular measurements and serum levels of sex hormones. However, there was no effect on serum cortisol. It was concluded from the present study that dietary supplementation of vitamin E has beneficial effects on the semen and hormones in male reproductive system.

Keywords: hormones, semen, teddy bucks, testicular measurements

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Authors: N. Isnaini, S. Wahjuningsih

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Fig fruit is the fruit of a tropical plant with content of flavanoids, vitamins A, C, and E which are antioxidants that effectively prevent and neutralize free radicals. This study was conducted to evaluate the supplementation of fig fruit extract in a physiological NaCl-based diluent on sperm motility and viability of native chicken semen after cooling. Semen was collected from 4 male mature chocks using massage method. Fresh semen evaluated for colour, pH, volume, concentration, mass motility, individual motility, life sperm and sperm abnormality. Semen was diluted with physiological NaCl-based extender supplemented with different levels of fig fruit extract (0, 10, 20 and 30 %) v/v with the ratio of 1 semen: 4 diluter. Semen used had mass motility of 2+ and motility of 70%. Immediately after dilution semen was stored in 3-5 °C and sperm motility and viability percentage were observed at 0, 12 and 24 h. The obtained data were analyze with Analysis of Variant (ANOVA) and Least Significant Difference were determined. The experiment was designed using completely random design (4 treatments and 10 replications). The results showed that the level of fig fruit extract had very significant effect (P < 0,01) on sperm motility and viability percentage in 0, 12 and 24 h of cooling. It can be concluded that the best fig fruit extract level for resulting optimal sperm motility and viability was 10%.

Keywords: chock, antioxidant, fig fruit extract, sperm

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Authors: Rida Pervaiz, Bushra Allah Rakha, Muhammad Sajjad Ansari, Shamim Akhter, Kainat Waseem, Sumiyyah Zuha, Tooba Javed

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Ring-necked pheasant (Phasianus colchicus) belongs to order Galliformes and family Phasianidae. It has been recognized as the most hunted bird due to its attractive colorful appearance and meat. Loss of habitat and hunting pressure has caused population fluctuations in the native range. Under these circumstances, this species can be conserved by employing ex-situ in vitro conservation techniques. Captive breeding, in combination with semen cryobanking is the most appropriate option to conserve/propagate this species without deteriorating the genetic diversity. Cryopreservation protocols of adequate efficiency are necessary to establish semen cryobanking for a species. Therefore, present study was designed to devise an efficient extender for cryopreservation of ring-necked pheasant semen. For this purpose, a range of extenders (Beltsville Poultry, red fowl, Lake, EK, Tselutin Poultry and Chicken semen extenders) were evaluated for cryopreservation of ring-necked pheasant semen. Semen collected from 10 cocks, diluted in the Beltsville Poultry (BPSE), Red Fowl (RFE), Lake (LE), EK (EKE), Tselutin Poultry (TPE) and Chicken Semen (CSE) extenders and cryopreserved. Glycerol (10%) was added to semen at 4°C, equilibrated for 10 min, filled in 0.5 mL French straws, kept over liquid nitrogen vapors for 10 min, cryopreserved in LN2 and stored. Sperm motility (%), viability (%), live/dead ratio (%), plasma membrane (%) and DNA Integrity (%) were evaluated at post-dilution, post-cooling, post-equilibration and post-thawing stage of cryopreservation. Sperm motility (83.8 ± 3.1; 81.3 ± 3.8; 73.8 ± 2.4; 62.5 ± 1.4), viability (79.0 ± 1.7; 75.5 ± 1.6; 69.5 ± 2.3; 65.5 ± 2.4), live/dead ratio (80.5 ± 5.7; 77.3 ± 4.9; 76.0 ± 2.7; 68.3 ± 2.3), plasma membrane (74.5 ± 2.9; 73.8 ± 3.4; 71.3 ± 2.3; 75.0 ± 3.4) and DNA integrity (78.3 ± 1.7; 73.0 ± 1.2; 68.0 ± 2.0; 63.0 ± 2.5) at all four stages of cryopreservation were recorded higher (P < 0.05) in red fowl extender compared to all experimental extenders. It is concluded that red fowl extender is the best extender for cryopreservation of ring-necked pheasant semen and can be used in establishing cryobank for ex situ conservation.

Keywords: ring-necked pheasant; extenders; cryopreservation; semen quality; DNA integrity

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Authors: Lourdita Llanto

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This study described and assessed the implementation of artificial insemination (AI) for cattle and carabaos in the Bicol Region, Philippines: Albay, Sorsogon and Camarines Sur. Three hundred respondents were interviewed. Results were analyzed using frequency counts, means, percentages and chi-square test. Semen samples from different stations were analyzed for motility, viability and morphology. T-test was used in semen quality evaluation. Provincial AI coordinators (PAIC) were male, averaging 59 years old, married, had college education, served in government service for 34 years, but as PAIC for 5.7 years. All had other designations. Mean AI operation was 11.33 years with annual support from the local government unit of Php76,666.67. AI technicians were males, married, with college education, and trained on AI. Problems were on mobility; inadequate knowledge of farmers in animal raising and AI; and lack of liquid nitrogen and frozen semen supply. There was 2.95 municipalities and breedable cattle/carabaos of 3,091.25 per AI technician. Mean number of artificially inseminated animals per AI technician for 2011 was 28.57 heads for carabaos and 8.64 heads for cattle. There was very low participation rate among farmers. Carabaos were 6.52 years with parity 1.53. Cattle were 5.61 years, with parity of 1.51. Semen quality significantly (p ≤ 0.05) deteriorated in normal and live sperm with storage and handling at the provincial and field stations. Breed, AI technicians practices and AI operation significantly affected conception rate. Mean conception rate was 57.62%.

Keywords: artificial insemination, carabao, parity, mother tanks, frozen semen

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Authors: Eva Tvrdá, Marek Halenár, Hana Greifová, Alica Mackovich, Faridullah Hashim, Norbert Lukáč

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Oxidative stress associated with semen cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Curcumin (CUR), a natural phenol isolated from Curcuma longa Linn. has been presented as a possible supplement for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of CUR on selected oxidative stress parameters in cryopreserved bovine semen. 20 bovine ejaculates were split into two aliquots and diluted with a commercial semen extender containing CUR (50 μmol/L) or no supplement (control), cooled to 4 °C, frozen and kept in liquid nitrogen. Frozen straws were thawed in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, and reactive oxygen species (ROS) generation was quantified by using luminometry. Superoxide generation was evaluated with the NBT test, and LPO was assessed via the TBARS assay. CUR supplementation significantly (P<0.001) increased the spermatozoa motility and provided a significantly higher protection against ROS (P<0.001) or superoxide (P<0.01) overgeneration caused by semen freezing and thawing. Furthermore, CUR administration resulted in a significantly (P<0.01) lower LPO of the experimental semen samples. In conclusion, CUR exhibits significant ROS-scavenging activities which may prevent oxidative insults to cryopreserved spermatozoa and thus may enhance the post-thaw functional activity of male gametes.

Keywords: bulls, cryopreservation, curcumin, lipid peroxidation, reactive oxygen species, spermatozoa

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Authors: Khosro Ghazvinian, Reza Narenji Sani, Touba Khodaiean, Melika Moezifar

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Many previous studies have reported that eCG was effective for completing spermatogenesis. In mice, eCG increased testes weight. In addition, Oxytocin (OT) was important in sperm transition and sperm motility in domestic animals. Peripheral circulation of OT also, was increased during sex incitement and ejaculation The objective of this study was to investigate the effect of IM injection of eCG and OT on semen characteristics in Zel rams in out of breeding season. Eighteen 3-year-old Zel adult rams were randomly divided into five equal groups (control and four treatment groups). 0.9% NaCl (1 ml) was injected IM into each ram in the control group, whereas eCG was administered IM at a single dose of 400 IU and 600 IU to each ram in the two eCG treatment groups and OT was administered IM at a single dose of 5 IU and 10 IU to each ram in the other two OT treatment groups. Semen samples were taken by an electroejaculator from all rams 10 min after the IM injection of 0.9% NaCl, eCG, or OT. eCG did not alter semen volume, and OT did not alter sperm motility or abnormal sperm, in comparison to the control values. Mass activity, sperm motility and total sperm number increased significantly in eCG group compared to the control group; and semen volume, mass activity, total sperm number of the OT treatment groups increased significantly compared to the control group. Exogenous 600 IU eCG and 10 IU OT increase mass activity, total sperm number, lived sperm and sperm concentration in Zel rams.

Keywords: eCG, oxytocine, semen characteristics, Zel Ram, nonbreeding season

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Authors: Muhammad Naveed Ali

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A study was conducted to determine the effects on the severity of pneumonia due to Mannheimia haemolytica and its relation with the fertility of bucks. A total of 12 crossbred Boer bucks of 3 treatment groups of equal number (4 goats per group, aged 12-14 months) were selected in this study. Group A was intranasally inoculated live M. haemolytica 1 × 105 (cfu). Group B was first immunized subcutaneously M. haemolytica killed vaccine (2 ml) two week before intranasal inoculation of M. haemolytica 1 × 105 (cfu). Group C was treated with normal saline (PBS) as control. Electro-ejaculator was used for semen collection once per week whilst scrotal circumference was measured before and after challenge. The semen volume, sperm concentration, sperm motility, live/dead percentage and morphology were evaluated. From the semen evaluation, goats in Group A exhibited significant decrease in the semen volume, sperm concentration, motility and live/dead sperm compared with vaccinated group B. The scrotal circumference was significantly decreased in group A compared to B. There were non-significant differences in scrotal circumferences of group B and C. The results suggested that M. haemolytica infection has negative effects on the fertility of Boer bucks.

Keywords: Boer bucks, Mannheimia haemolytica, semen evaluation, vaccination

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Authors: Sri Rahayu, M. Dwi Susan, Aris Soewondo, W. M. Agung Pramana

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Semen is an organic fluid (seminal plasma) that contain spermatozoa. Proteins are one of the major seminal plasma components that modulate sperm functionality, influence sperm capacitation and maintaining the stability of the membrane. Semen freezing is a procedure to preserve sperm cells. The process causes decrease in sperm viability due to temperature shock and oxidation stress. Oxidation stress is a disturbance on phosphorylation that increases ROS concentration, and it produces lipid peroxide in spermatozoa membrane resulted in high MDA (malondialdehyde) concentration. The objective of this study was to examine the effect of freezing on protein and MDA profile of bovine sperm cell and seminal plasma after freezing. Protein and MDA of sperm cell and seminal plasma were isolated from 10 sample. Protein profiles was analyzed by SDS PAGE with separating gel 12,5 %. The concentration of MDA was measured by spectrophotometer. The results of the research indicated that freezing of semen cause lost of the seminal plasma proteins with molecular with 20, 10, and 9 kDa. In addition, the result research showed that protein of the sperm (26, 10, 9, 7, and 6 kDa) had been lost. There were difference MDA concentration of seminal plasma and sperm cell were increase after freezing. MDA concentration of seminal plasma before and after freezing were 2.2 and 2.4 nmol, respectively. MDA concentration of sperm cell before and after freezing were 1,5 and 1.8 nmol, respectively. In conclusion, there were differences protein profiles of spermatozoa before and after semen freezing and freezing cause increasing of the MDA concentration.

Keywords: MDA, semen freezing, SDS PAGE, protein profile

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Authors: Aravind Bagade Shankaranarayana, Parampalli Geetha, Pallavi Gupta

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Objective: This study intends to evaluate sexual dysfunction qualitatively and quantitatively in males suffering from primary obesity through a single centered, observational study. Design and Methods: Sexual function of 33 obese males from the outpatient department of the hospital was assessed using IIEF questionnaire and semen analysis and the results were assessed for statistical significance. Results: A varying degree of sexual dysfunction was observed in four out of five areas of sexual functioning viz. erectile function (p<0.02), orgasmic function (p<0.02), sexual desire (p<0.08) and overall satisfaction (p<0.000) in obese individuals. Statistically significant dysfunction was not observed in intercourse satisfaction. Semen analysis was normal in 19 individuals (63.3%) and abnormal in 11 individuals (36.7%), with statistically insignificant p value 0.144, suggesting mild to moderate variation in semen parameters. Conclusions: Varying degree of sexual dysfunction is present in obese males, suggesting that obesity has a possible role in reducing the quality of sexual functioning in males as indicated in the classical Ayurvedic literature.

Keywords: erectile dysfunction, krucchra vyavaya, obesity, sthoulya

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Authors: Abdurzag Kerban, Mostfa M. Abou-Ahmed, Abdelrof M. Ghallab, Mona H. Shaker

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Preservation of semen had a major impact on sheep genetic breeding. The aim of this study was to evaluate the effect of protected fat, probiotic and zinc-enriched diets on semen freezability. Twenty two Barki rams were randomly assigned into four groups; Group I (n=5) was fed the basal diet enriched with 3.7% of dry fat/kg concentration/day, Group II (n=5) was fed a basal diet-enriched with 10gm of probiotic /head/day, Group III (n=6) was fed on the basal diet enriched with 100 ppm of 10% zinc chelated with methionine/kg dry matter/day and Group IV (n=6) was served as control. A pool of three to four ejaculates were pooled from rams within a period of ten weeks. Semen was diluted in egg yolk-Tris diluent and processed in 0.25 ml straw. Motility was evaluated after dilution, before freezing and post-thawing at 0, 1, 2 and 3 hour incubation. Viability index, acrosome integrity and leakage of intracellular enzymes (Aspartat aminotransferase and Alkline phosphatase) were also evaluated. Spermatozoa exhibited highly significant (P<0.01) percentages of motility at 0, 1, 2, and 3 hours incubation after thawing, viability index and acrosome integrity in rams fed a diet enriched with protected fat and zinc groups as compared with probiotic and control groups. Also, the mean value of extracellular leakage of AST was significantly lower in fat and zinc group as compared with probiotic and control groups. In conclusion, semen freezability was improved in animals fed a diet fortified with fat and zinc with no significant improvement in animals fed the probiotic-enriched diet.

Keywords: Barki ram semen, freezing, straw, feed additives

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Authors: M. H. Moreira da Silva, L. Valadao, F. Moreira da Silva

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In the present study, the fertilizing capacity of bovine spermatozoa was evaluated before and after its cryopreservation. For this, the testicles of 100 bulls slaughtered on Terceira Island were dissected, the epididymal tails were separated, and semen was recovered by the flotation method and then evaluated by phase contrast microscopy and by flow cytometry. For phase contrast microscopy, a drop of semen was used to evaluate the percentage of motile spermatozoa (from 0 to 100%) and motility (from 0 to 5). After determining the concentration and the abnormal forms, semen was diluted to a final concentration of 50 x 106 spz/ml and evaluated by flow cytometer for membrane and acrosome integrity using the conjugation of fluorescent probes propidium iodide (PI) and Arachis hypogea agglutinin (FITC-PNA). Freezing was carried out in a programmable semen freezer, using 0.25 ml straws, in a total of 20 x 106 viable sperm per straw with glycerol as a cryoprotectant in a final concentration of 0.58 M. It was observed that, on average, a total of 7.25 ml of semen was collected from each bull. The viability and vitality rates were respectively 83.22 ± 7.52% and 3.8 ± 0.4 before freezing, decreasing to 58.81 ± 11.99% and 3.6 ± 0.6, respectively, after thawing. Regarding cytoplasmic droplets, it was observed that a high percentage of spermatozoa had medial cytoplasmic droplets (38.47%), with only 3.32% and 0.15% presenting proximal and distal cytoplasmic drops, respectively. By flow cytometry, it was observed that before freezing, the percentage of sperm with the damaged plasma membrane and intact acrosome was 3.61 ± 0.99%, increasing slightly to 4.21 ± 1.86% after cryopreservation (p<0.05). Regarding spermatozoa with damaged plasma membrane and acrosome, the percentage before freezing was 3.37±1.87%, increasing to 4.34 ±1.16% after thawing, and no significant differences were observed between these two values. For the percentage of sperm with the intact plasma membrane and damaged acrosome, this value was 2.04 ± 2.34% before freezing, decreasing to 0.89 ± 0.48% after thawing (p<0.05). The percentage of sperm with the intact plasma membrane and acrosome before freezing was 90.99±2.75%, with a slight decrease to 90.57±3.15% after thawing (p<0.05). From this study, it can be clearly concluded that, after the slaughtering of bulls, the spermatozoa can be recovered from the epididymis and cryopreserved, maintaining an excellent rate of sperm viability and quality after thawing.

Keywords: bovine semen, epididymis, cryopreservation, fertility assessment

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Authors: K. A. El-Battawy, E. Brannas

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Cryopreservation of sperm causes damages and adversely affected sperm motility and viability resulting in lower hatching rates. The aim of this study is to determine whether propolis has potential protective effect on cryopreservation and fertilization ability of spermatozoa of Salvelinusalpinus. The extenders were prepared by using simple glucose solution (0.3 M glucose) to which 10% Me2SO added with different levels of propolis (0.4, 0.8 and 1 mg/ ml) and 10% egg yolk (as a control without propolis). The pooled semen samples diluted at the ratio of 1:3 by the extenders were subjected to cryopreservation. The percentage and duration of motility and fertilization tests of cryopreserved sperm samples have been done immediately after thawing and compared with control and fresh semen. The extenders containing propolis showed higher percentage motility and motility duration than control group (P < 0.05). Especially the group II (0.8 mg/ ml propolis) and the group III (1 mg/ ml propolis) showed significant positive effects on both post thaw motility and hatching ability. In conclusion, this study confirms that the propolis is an appropriate cryoptrotective agent in fish semen and it maintained the integrity of the spermatozoa during the cryopreservation process.

Keywords: propolis, arctic charr, semen, cryopreservation

Procedia PDF Downloads 287

Authors: Ashraf Shebl, Bassam Garah, Radah Youssef

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Sexual assault is usually a hidden crime where the only witnesses are the victim and the assailant. For a variety of reasons, even the victim may be unable to provide a detailed account of the assault or the identity of the perpetrator. Often the case history deteriorates into one person’s word against another. With such limited initial information, the physical and biological evidence collected from the victim, from the crime scene, and from the suspect will play a pivotal role in the objective and scientific reconstruction of the events in question. The aim of work is to examine whether DNA profiles could be recovered from repeated washed clothes after contaminated by semen stains. Fresh semen about 1ml. ( <1 h old) taken from donor was deposited on four types of clothes (cotton, silk, polyester, and jeans). Then leave to dry in room temperature and washed by washing machine at temperature (30°C-60°C) and by hand washing. Some items of clothing were washed once, some twice and others three times. DNA could be extracted from some of these samples even after multiple washing. This study demonstrates that complete DNA profiles can be obtained from washed semen stains on different types of clothes, even after many repeated washing. These results indicated that clothes of the victims must be examined even if they were washed many times.

Keywords: sexual assault, DNA, persistence, clothes

Procedia PDF Downloads 198

Authors: Asaduzzaman Rimon, Pankaj Kumar Jha, Abdullah Al Mansur, Mohammad Mofizul Islam, Nasrin Sultana Juyena, Farida Yeasmin Bari

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This study was conducted to determine the effects of multi-species skim milk based extenders on sperm quality at 5ºC with the advancement of preservation time. Altogether forty ejaculates, 8 ejaculates for each of the 5 home-made semen extenders: cow skim milk (CSM), goat skim milk (GSM), sheep skim milk (SSM), buffalo skim milk (BSM) and commercial dried skim milk (CDSM) were examined for motility, plasma membrane integrity and normal morphology % of sperm at 0, 24, 48, 72, 96 and 120 hours, respectively. Sperm motility was significantly decreased (P < 0.05) with the increase of preservation time. There were no significant difference in motility % among CSM (84.0±1.4, 82.3±2.1), GSM (84.5±1.0, 82.5±0.6) and CDSM (85.0±80.3±1.3) extenders at 0 and 24 hours, respectively. However, the motility in GSM extender was significantly higher than BSM, SSM and CDSM extender at 48, 72, 96 and 120 hours. The plasma membrane integrity % at 0 hour had no significant difference among the extenders. But, the plasma membrane integrity % in GSM (84.3±0.9, 81.8±1.3, 78.0±2.2, 74.8±0.5, 72.0±1.4) and CSM (82.8±0.5, 80.8±1.0, 78.0±1.4, 73.5±1.7, 70.3±0.5) extenders were significantly higher than BSM (81.0±1.4, 76.3±2.5, 72.5±1.7, 63.8±2.5, 54.0±4.6), SSM (78.5±1.5, 75.0±1.6, 71.5±2.4, 64.3±1.7, 56.5±2.4) and CDSM extenders (78.3±2.4, 75.8±3.9, 72.5±3.3, 64.8±1.0, 60.5±3.3) at 24, 48, 72, 96 and 120 hours, respectively. The sperm morphology % had no significant difference at 0 hour among the extenders but were significantly higher in GSM (83.0±0.8, 81.3±1.5, 79.3±1.3, 73.0±2.2, 70.3±1.3) and CSM (81.5±1.7, 79.3±1.5, 75.8±1.5, 70.3±1.3, 66.3±1.5) than BSM (79.0±1.2, 75.0±1.4, 69.5±1.7, 64.5±3.1, 56.8±2.2), SSM (79.8±1.3, 76.8±2.1, 71.3±3.0, 66.0±2.7, 60.3±4.5) and CDSM (80.0±1.6, 77.0±2.2, 72.0±2.5, 66.3±2.5, 62.0±4.0) extenders at 24, 48, 72, 96 and 120 hours, respectively. The motility, plasma membrane integrity and normal morphology % of sperm had shown no significant difference between GSM and CSM but were found to be higher in GSM extenders. In the end, we concluded from the above study that the goat milk based extenders (GSM) had optimum sperm preserving quality. However, further studies are required to validate followed by fertility rate.

Keywords: chilled semen, indigenous ram, multi-species skim milk based extenders, preservation

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Authors: Meenu Joshi, Natalie Naidoo, Farzeen Kader

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Identification of body fluids is an essential step in forensic investigation to aid in crime reconstruction. Tissue-specific differentially methylated regions (tDMRs) of the human genome can be targeted to be used as biomarkers to differentiate between body fluids. The present study was undertaken to establish the methylation status of potential tDMRs in blood, semen, saliva, and vaginal fluid by using methylation-specific PCR (MSP) and bisulfite sequencing (BS). The methylation statuses of 3 potential tDMRS in genes ZNF282, PTPRS, and HPCAL1 were analysed in 10 samples of each body fluid. With MSP analysis, the ZNF282, and PTPRS1 tDMR displayed semen-specific hypomethylation while HPCAL1 tDMR showed saliva-specific hypomethylation. With quantitative analysis by BS, the ZNF282 tDMR showed statistically significant difference in overall methylation between semen and all other body fluids as well as at individual CpG sites (p < 0.05). To evaluate the effect of environmental conditions on the stability of methylation profiles of the ZNF282 tDMR, five samples of each body fluid were subjected to five different forensic simulated conditions (dry at room temperature, wet in an exsiccator, outside on the ground, sprayed with alcohol, and sprayed with bleach) for 50 days. Vaginal fluid showed highest DNA recovery under all conditions while semen had least DNA quantity. Under outside on the ground condition, all body fluids except semen showed a decrease in methylation level; however, a significant decrease in methylation level was observed for saliva. A statistical significant difference was observed for saliva and semen (p < 0.05) for outside on the ground condition. No differences in methylation level were observed for the ZNF282 tDMR under all conditions for vaginal fluid samples. Thus, in the present study ZNF282 tDMR has been identified as a novel and stable semen-specific hypomethylation marker.

Keywords: body fluids, bisulphite sequencing, forensics, tDMRs, MSP

Procedia PDF Downloads 163

Authors: Diana Peninger

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Over 10% of couples in the U.S. have infertility problems, with roughly 40% traceable to the male partner. Yet, little attention has been given to improving men’s contribution to the conception process. One solution that is showing promise in increasing conception rates for IVF and other assisted reproductive technology treatments is a first-of-its-kind semen collection that has been engineered to mitigate sperm damage caused by traditional collection methods. Patients are able to collect semen at home and deliver to clinics within 48 hours for use in fertility analysis and treatment, with less stress and improved specimen viability. This abstract will share these findings along with expert insight and tips to help attendees understand the key role sperm collection plays in addressing and treating reproductive issues, while helping to improve patient outcomes and success. Our research was to determine if male reproductive outcomes can be increased by improving sperm specimen health with a focus on technology. We utilized a redesigned semen collection cup (patented as the Device for Improved Semen Collection/DISC—U.S. Patent 6864046 – known commercially as a ProteX) that met a series of physiological parameters. Previous research demonstrated significant improvement in semen perimeters (motility forward, progression, viability, and longevity) and overall sperm biochemistry when the DISC is used for collection. Animal studies have also shown dramatic increases in pregnancy rates. Our current study compares samples collected in the DISC, next-generation DISC (DISCng), and a standard specimen cup (SSC), dry, with the 1 mL measured amount of media and media in excess ( 5mL). Both human and animal testing will be included. With sperm counts declining at alarming rates due to environmental, lifestyle, and other health factors, accurate evaluations of sperm health are critical to understanding reproductive health, origins, and treatments of infertility. An increase in the health of the sperm as measured by extensive semen parameter analysis and improved semen parameters stable for 48 hours, expanding the processing time from 1 hour to 48 hours were also demonstrated.

Keywords: reprodutive, sperm, male, infertility

Procedia PDF Downloads 129