Search results for: semen freezing

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: 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: 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

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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: 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: 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: 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: 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: 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

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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: Thanh Phuong Doan, Narueporn Sutanthavibul

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The influence of freezing cycle on influenza haemagglutinin (HA) conformational stability was investigated in terms of freezing rates and formulation compositions. The results showed that appropriate HA conformation could be evaluated using circular dichroism (CD) spectroscopy with HA concentration of greater than 0.09 mg/ml. The intermediate freezing rate of approximately 1.0oC/min preserved the original HA conformation better than at slow freezing rate (0.5oC/min) and rapid freezing rate (2.6oC/min). The changes in CD spectra of the secondary HA structure were more pronounced than those of the tertiary HA structure during the evaluation. Additionally, the formulations, which resulted in the highest conformational stability were found to have sucrose present in the composition. As opposed to when only glycine was used, the stability of HA conformation was poor.

Keywords: freezing, haemagglutinin, influenza, circular dichroism

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Authors: Leila Ben Haj Said, Sihem Bellagha, Karim Allaf

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The present study deals with the dehydrofreezing assisted by instant controlled pressure drop (DIC) treatment of apple fruits. Samples previously dehydrated until different water contents (200, 100, and 30% dry basis (db)) and DIC treated were frozen at two different freezing velocities (V+ and V-), depending on the thermal resistance established between the freezing airflow and the sample surface. The effects of sample water content (W) and freezing velocity (V) on freezing curves and characteristics, exudate water (EW) and texture variation were examined. Lower sample water content implied higher freezing rates, lower initial freezing points (IFP), lower practical freezing time (PFT), and lower specific freezing time (SFT). EW (expressed in g exudate water/100 g water in the product) of 200% and 100% db apple samples was approximately 3%, at low freezing velocity (V-). Whereas, it was lower than 0.5% for apple samples with 30% db water content. Moreover, the impact of freezing velocity on EW was significant and very important only for high water content samples. For samples whose water content was lower than 100% db, firmness (maximum puncture force) was as higher as the water content was lower, without any insignificant impact of freezing velocity.

Keywords: dehydrofreezing, instant controlled pressure drop DIC, freezing time, texture

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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: 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: 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: Hamid Reza Khodaei, Behnaz Mahdavi, Alireza Banitaba

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Nitric oxide (NO) plays an important role in all sexual activities of animals. It is made in body from NO syntheses enzyme and L-arginin molecule. NO can make band with sulfur-iron complexes and due to production of steroid sexual hormones related to enzymes which have this complex, NO can change the activity of these enzymes. NO affects many cells including endothelial cells of veins, macrophages and mast cells. These cells are found in testis leydig cells and therefore are important source of NO in testis tissue. Minimizing damages to sperm at the time of sperm freezing and thawing is really important. The goal of this study was to determine the function of NO before freezing and its effects on quality and viability of sperms after thawing and incubation. 4 Holstein bulls were selected from the age of 4, and artificial insemination was done for 3 weeks (2 times a week). Treatments were 0, 10, 50 and 100 nm of sodium nitroprusside (SNP). Data analysis was performed by SAS98 program. Also, mean comparison was done using Duncan's multiple ranges test (P<0.05). Concentrations used were found to increase motility and viability of spermatozoa at 1, 2 and 3 hours after thawing significantly (P<0.05) but there was no significant difference at zero time. SNP levels reduced the amount of lipid peroxidation in sperm membrane, increased acrosome health and improved samples membranes especially in 50 and 100 nm treatments. According to results, adding SNP to semen diluents increases motility and viability of spermatozoa. Also, it reduces lipid peroxidation in sperm membrane and improves sperm function.

Keywords: sperm motility, nitric oxide, lipid peroxidation, spermatozoa

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

Abstract:

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: Leila Karshenas, Hamid Reza Khodaei, Behnaz Mahdavi

Abstract:

We know that nitric oxide (NO) plays an important role in all sexual activities of animals. It is made in body from NO synthase enzyme and L-arginin molecule. NO can bound with sulfur-iron complexes and because production of steroid sexual hormones is related to enzymes which have this complex, NO can change the activity of these enzymes. NO affects many cells including endothelial cells of veins, macrophages and mast cells. These cells are found in testis leydig cells and therefore are important source of NO in testis tissue. Minimizing damages to sperm at the time of sperm freezing and thawing is really important. The goal of this study was to determine the function of NO before freezing and its effects on quality and viability of sperms after thawing and incubation. 4 Holstein bulls were selected from the age of 4, and artificial insemination was done for 3 weeks (2 times a week). Treatments were 0, 10, 50 and 100 nm of sodium nitroprusside (SNP). Data analysis was performed by SAS98 program. Also, mean comparison was done using Duncan's multiple ranges test (P<0.05). Concentrations used was found to increase motility and viability of spermatozoa at 1, 2 and 3 hours after thawing significantly (P<0.05), but there was no significant difference at zero time. SNP levels reduced the amount of lipid peroxidation in sperm membrane, increased acrosome health and improved sample membranes especially in 50 and 100 nm treatments. According to results, adding SNP to semen diluents increases motility and viability of spermatozoa. Also, it reduces lipid peroxidation in sperm membrane and improves sperm function.

Keywords: sperm motility, nitric oxide, lipid peroxidation, spermatozoa

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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: 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: 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

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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

Procedia PDF Downloads 528

Authors: Martyna Ekiert, Andrzej Mlyniec

Abstract:

Tendons are the biological structures, which primary function is to transfer force generated by muscles to the bones. Unfortunately, damages of tendons are also one of the most common injuries of the human musculoskeletal system. For the most severe cases of tendon rupture, such as the tear of calcaneus tendon or anterior cruciate ligament of the knee, a surgical procedure is the only possible way of full recovery. Tendons used as biological grafts are usually subjected to the process of deep freezing and subsequent thawing. This, in particular for multiple freezing/thawing cycles, may result in changes of tendon internal structure causing deterioration of mechanical properties of the tissue. Therefore, studies on the influence of freezing on tendons biomechanics, including internal water content in soft tissue, seems to be greatly needed. An experimental study of the influence of freezing on mechanical properties of the tendon was performed on fascicles samples dissected form bovine flexor tendons. The preparation procedure was performed with the presence of 0.9% saline solution in order to prevent an excessive tissue drying. All prepared samples were subjected to the different number of freezing/thawing cycles. For freezing part of the protocol we used -80°C temperature while for slow thawing we used fridge temperature (4°C) combined with equalizing temperatures in the standard state (25°C). After final thawing, the mechanical properties of each sample was examined using cyclic loading test. Our results may contribute for better understanding of negative effects of soft tissues freezing, resulting from abnormal thermal expansion of water. This also may help to determine the limit of freezing/thawing cycles disqualifying tissue for surgical purposes and thus help optimize tissues storage conditions.

Keywords: freezing, soft tissue, tendon, bovine fascicles

Procedia PDF Downloads 177

Authors: A. Şahin Zaimoglu

Abstract:

A number of studies have been conducted recently to investigate the influence of randomly oriented fibers on some engineering properties of cohesive and cohesionless soils. However, few studies have been carried out on freezing-thawing behavior of fine-grained soils modified with discrete fiber inclusions and additive materials. This experimental study was performed to investigate the effect of randomly distributed polypropylene fibers (PP) and some additive materials [e.g.., borogypsum (BG), fly ash (FA) and cement (C)] on freezing-thawing durability (mass losses) of a fine-grained soil for 6,12 and 18 cycles. The Taguchi method was applied to the experiments and a standard L9 orthogonal array (OA) with four factors and three levels were chosen. A series of freezing-thawing tests were conducted on each specimen. 0-20 % BG, 0-20 % FA, 0-0.25 % PP and 0-3 % of C by total dry weight of mixture were used in the preparation of specimens. Experimental results showed that the most effective materials for the freezing-thawing durability (mass losses) of the samples were borogypsum and fly ash. The values of mass losses for 6, 12 and 18 cycles in optimum conditions were 16.1%, 5.1% and 3.6%, respectively.

Keywords: freezing-thawing, additive materials, reinforced soil, optimization

Procedia PDF Downloads 269

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

Abstract:

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

Procedia PDF Downloads 109

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

Abstract:

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

Procedia PDF Downloads 333

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

Abstract:

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

Procedia PDF Downloads 367

Authors: Muhammad Naveed Ali

Abstract:

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

Procedia PDF Downloads 119

Authors: Azam Fazelpour, Saeed R. Dehghani, Vlastimil Masek, Yuri S. Muzychka

Abstract:

The study uses an image analysis technique employing thermal imaging to measure the percentage of areas with various temperatures on a freezing surface. An image segmentation method using threshold values is applied to a sequence of image recording the freezing process. The phenomenon is transient and temperatures vary fast to reach the freezing point and complete the freezing process. Freezing salt water is subjected to the salt rejection that makes the freezing point dynamic and dependent on the salinity at the phase interface. For a specific area of freezing, nucleation starts from one side and end to another side, which causes a dynamic and transient temperature in that area. Thermal cameras are able to reveal a difference in temperature due to their sensitivity to infrared radiance. Using Experimental setup, a video is recorded by a thermal camera to monitor radiance and temperatures during the freezing process. Image processing techniques are applied to all frames to detect and classify temperatures on the surface. Image processing segmentation method is used to find contours with same temperatures on the icing surface. Each segment is obtained using the temperature range appeared in the image and correspond pixel values in the image. Using the contours extracted from image and camera parameters, stratified areas with different temperatures are calculated. To observe temperature contours on the icing surface using the thermal camera, the salt water sample is dropped on a cold surface with the temperature of -20°C. A thermal video is recorded for 2 minutes to observe the temperature field. Examining the results obtained by the method and the experimental observations verifies the accuracy and applicability of the method.

Keywords: ice contour boundary, image processing, image segmentation, salt ice, thermal image

Procedia PDF Downloads 282

Authors: Farnaz Amidi Fazli

Abstract:

After long period of human experience about food processing from raw eating to canning of food in the last century now it is time to use novel technologies which are sometimes completely different from common technologies. It is possible to decontaminate food without using heat or the foods are stored without using cold chain. Pulsed electric field (PEF) processing is a non-thermal method of food preservation that uses short bursts of electricity, PEF can be used for processing liquid and semi-liquid food products. PEF processing offers high quality fresh-like liquid foods with excellent flavor, nutritional value, and shelf-life. High pressure processing (HPP) technology has the potential to fulfill both consumer and scientific requirements. The use of HPP for over 50 years has found applications in non-food industries. For food applications, ‘high pressure’ can be generally considered to be up to 600 MPa for most food products. After years, freezing has its high potential to food preservation due to new and quick freezing methods. Foods which are prepared by this technology have more acceptability and high quality comparing with old fashion slow freezing. Thus, quick freezing has further been adopted as a widespread commercial method for long-term preservation of perishable foods which improved both the health and convenience of everyone in the industrialised countries. Above parameters are achieved by Fluidised-bed freezing systems, freezing by immersion and Hydrofluidisation on the other hand new thawing methods like high-pressure, microwave, ohmic, and acoustic thawing have a key role in quality and adaptability of final product.

Keywords: quick freezing, thawing, high pressure, pulse electric, hydrofluidisation

Procedia PDF Downloads 290