Percentage Of Live Spermatozoa Research Articles

Canine reproductive ultrasound examination for predicting future sperm quality.

The reproductive potential of male animals is commonly evaluated using a breeding soundness examination incorporating B-mode ultrasound examination of the testes and recently Doppler ultrasound examination of the testicular arteries. These techniques may detect testicular normality or pathology, and while some measured parameters are associated with semen quality at the time of ultrasound examination, few studies have investigated the relationship with future semen quality. We hypothesized that B-mode and Doppler ultrasound measurements would correlate with future semen quality. Within two studies, we investigated the relationship between ultrasound measured testicular volume, testicular echogenicity, testicular homogeneity, subjective assessment of the testicular parenchyma, testicular artery resistance index, and pulsatility index with subsequent semen quality. Fifty-five normal fertile dogs of which 29 had stable semen quality and 26 had a subsequent decline in semen quality were examined during a 6-month period commencing 62days after the ultrasound examination. Statistical analysis showed that no ultrasound parameters were predictive of future total sperm output or percentage live normal sperm. However, mean testicular echogenicity was positively related to future sperm motility (t=2.202, p=.039). We conclude that quantitative ultrasound assessment of the appearance of the testicular parenchyma has potential for the evaluation of future semen quality in dogs.

Prophylactic effect of Nigella sativa against lead acetate induced changes in spermiogram, reproductive hormones and gonadal histology of rats.

Aim:This study was designed to evaluate the prophylactic effect of Nigella sativa (NS) treatment on toxic effects induced by lead acetate (LA) on the reproductive hormones, spermiogram and gonadal histology of rats.Materials and Methods:A total of 20 Sprague-Dawley rats were divided into four groups of five rats each. Group 1 (negative control [NC]) was the NC and was given distilled water, Group 2 served as the positive control (PC) and was administered 10 mg/kg/day of LA per overall survival (OS), Group 3 (T1) was administered 200 mg/kg/daily of NS per OS for a month, and Group 4 (T2) was pretreated with 200 mg/kg/daily of NS per OS for 1 month, followed by 10 mg/kg/daily of LA alone per OS for another. The rats were euthanized at the end of the experimental period for collection of blood and the right caudal epididymis and testis. Serum was used for determination of reproductive hormones by using radioimmunoassay kits. The epididymal segment was cut and homogenized in phosphate-buffered saline, and the homogenate was used for determination of the spermiogram parameters such as sperm concentration, sperm viability, percentage of live sperm, motility and abnormality. Both the epididymis and testis were fixed in 10% buffered formalin for histological processing.Results:The sperm concentration, general, and individual motilities were higher (p<0.05) in the NC and T1 animals, while the T2 had intermediate and the PC had lower (p<0.05) values of each parameter. The percentage sperm viability was higher (p<0.05) in the T1 and lower (p<0.05) in the PC group. However, percentage abnormality was lower in T1, comparable in NC and T2, and higher (p<0.05) in PC. Spermatogenic cell population and epididymal sperm reserve (ESR) were optimal in control and pretreated animals, while the PC had lower spermatids and ESR. The concentration of estradiol (EST) was lower (p<0.05) in the PC and T2, while leuteinizing hormone (LH) concentration was lower (p<0.05) in the PC, and comparable (p>0.05) between control and T2. The concentration of follicle-stimulating hormone (FSH) was comparable (p>0.05) in all groups, while testosterone (TS) hormone concentration was lower (p<0.05) in the PC and higher in the control and T1 groups.Conclusion:This study showed the preventive effects of NS administration against alterations in reproductive hormnes, sperm parameters and gonadal histology caused by LA in rats.

Determination of male-specific gene expression of White Lamphun cattle semen treated by monoclonal antibodies

In the present study, male-specific monoclonal antibodies, P1C2B8 and P1C2C9, were used for sperm sexing via agglutination and cytotoxic reactions in White Lamphun cattle semen no. 862, no. 863 and no. KL. Live sperm percentage was used to find out an optimum dilution of antibody in agglutination and cytotoxic reactions and multiplex real time polymerase chain reaction was used to determine male-specific gene expression in each optimum dilution. In agglutination treatment, the results showed that all cattle had the same optimum antibody dilution while in cytotoxicity treatment, each cattle had different optimum antibody dilution. The results from multiplex real time polymerase chain reaction also showed that in both agglutination and cytotoxicity treatments, male- specific gene expression in semen samples was lower in cattle no. 862, no. 863 and no. KL when compared to control treatment. From these results, it was implied that antibodies P1C2B8 and P1C2C9 could alter the sperm sex ratio by reducing Y sperm in White Lamphun cattle semen.

Influence of different anaesthetic protocols over the sperm quality on the fresh, chilled (4°C) and frozen-thawed epididymal sperm samples in domestic dogs.

This study assessed the influence of three different anaesthetic protocols on semen quality obtained from the epididymis. Sixty male dogs undergoing to routine sterilization were assigned to three anaesthetic protocols: thiopental group (TG, n=20), propofol group (PG, n=20) and ketamine-dexmedetomidine group (KDG, n=20). Immediately after orchidectomy, the cauda epididymides and vas deferent ducts were isolated and then a retrograde flushing was performed to collect spermatozoa. In experiment 1, after the initial evaluation of the semen (sperm concentration, sperm motility and the percentages of live spermatozoa, abnormal spermatozoa and acrosome membrane integrity), semen samples were diluted in Tris-glucose-egg yolk extender and chilled for 48hr, and the sperm motility was assessed at 6, 24 and 48hr. In experiment 2, semen samples were diluted in Tris-glucose-egg yolk extender and chilled for 24hr, and then samples were frozen in two extenders with different glycerol concentrations, to reach a final concentration of 50-100×10(6) spermatozoaml(-1) , 20% egg yolk, 0.5% Equex and 4% and 5% glycerol, respectively. Mean values of total sperm concentration, sperm viability and the percentages of intact acrosome and abnormal spermatozoa were not significantly different between experimental groups, and therefore, the anaesthetic protocols assessed did not affect sperm parameters mentioned above. However, our study confirmed a detrimental effect of the use of thiopental (TG) over the total sperm motility (p<0.05) and progressive sperm motility (p<0.05) of the fresh and chilled epididymal sperm samples. The anaesthetic protocols including the application of propofol or ketamine-dexmedetomidine can be used to recover sperm in domestic canids without significant changes in sperm quality compared when semen is collected routinely and these techniques could be applicable to endangered wild canids.

Caspase 3 Activity and Lipoperoxidative Status in Raw Semen Predict the Outcome of Cryopreservation of Stallion Spermatozoa.

Stallion-to-stallion variability in the quality of cryopreserved ejaculates postthaw affects the commercial acceptability of frozen semen and thus is a major constraint for the equine industry. In recent years, the molecular mechanisms associated with sperm damage during cryopreservation have become better understood. Identification of the freezability of the ejaculates before the freezing process is initiated will have a major impact on the equine industry. We studied three markers of oxidative stress in sperm, including 8-iso-PGF2alpha, 8-OH guanosine, and 4-hydroxynonenal (4-HNE); the presence of active caspase 3; and their changes after sperm cryopreservation. Although 4-HNE levels increased after cryopreservation (from 7% to 33%, P < 0.001), 8OH-guanosine and 8-ISO-PGF2alpha levels decreased after cryopreservation (from 130 to 35 arbitrary fluorescence units, P < 0.01, and from 1280 to 1233, P < 0.01, respectively). Postthaw sperm quality was classified as poor, average, or good using the 25th and 75th percentiles of all assays of sperm quality studied (motility, velocity, membrane functionality, and thiol content) as thresholds. Using these values, a sperm postthaw quality index was proposed. Receiver operating characteristic curves and the Youden J statistic were used to investigate the value of the measured parameters in fresh sperm as predictors of potential freezability. Using these techniques, we identified markers of bad freezers (percentages of caspase 3-positive dead sperm [area under the curve (AUC) = 0.820, P < 0.05] and percentages of caspase 3- and 4-HNE-positive sperm [AUC = 0.872, P < 0.05]) and good freezers (percentages of caspase 3-negative live sperm [AUC = 0.815, P < 0.05], percentages of live sperm with high thiol content [AUC = 0.907, P < 0.01], and percentages of 8-ISO-PGF2alpha-positive sperm [AUC = 0.900, P < 0.01]. Moreover, we described for the first time the presence of 8-ISO-PGF2alpha in stallion spermatozoa and revealed the importance of considering different markers of oxidative stress.

Effect of supplementation of feed with Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls.

BackgroundThe current study was designed to investigate the effect of supplementation of Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls.MethodsIn this study, 12 adult healthy bulls kept at the Semen Production Unit, Qadirabad district Sahiwal, were used. These bulls were divided into three equal groups, A, B and C. Group A was kept as control, while in groups B and C supplementation of feed was provided by using flaxseed oil @125 ml/day and 250 ml/day,respectively for 12 weeks. Two ejaculates per animal were collected at 0 day then 5th, 6th, 7th, 8th, 9th, 10th, 11th and 12th week of treatment. In this way a total 216 samples were taken, and each semen sample was evaluated for color, volume, mass activity, percent motility, sperm cell concentration per ml, percentage of live sperm, and plasma membrane integrity. Libido of bulls was also evaluated before every collection.ResultsAnalysis of data revealed that these parameters were significantly (P < 0.01) increased in flax oil treated animals as compared to control.ConclusionIt was concluded from the present study that flax seed oil has beneficial effects on reproductive health of buffalo bull.

Toxicological assessment of Diplazium esculentum on the reproductive functions of male Swiss albino mouse

Context: Diplazium esculentum, a commonly consumed seasonal vegetable, has been reported to have some pathological effects in some animals. But, its effect on the male reproductive function has not yet been studied. Objective: To investigate the effects of boiled D. esculentum (BDE), the form which human consumes, on male reproductive functions of Swiss albino mice. Materials and methods: Male (120 in no.) and female (80 in no.) Swiss albino mice (6–8 weeks of age) were fed orally with 80, 160 and 320 mg/kg bw of BDE within a span of 180 d. After the treatment, body weight, absolute- and relative-testis weight, relative-weight of other organs, their biochemical parameters, hypo-osmotic swelling test (HOST) of spermatozoa, testis histology and fertility and fecundity tests were performed to justify the toxic effects of D. esculentum on male reproductive functions. Results: Significant dose- and time-dependent decreases were observed in body weight, absolute- and relative-testis weight, relative-weights of other organs and their biochemical parameters, percentage of live spermatozoa and percentage of fertility and fecundity in BDE fed mice. Significant decreases were observed in diameter, perimeter and area of the seminiferous tubules of mice treated for 180 d. The percentage of empty seminiferous tubules was increased significantly in BDE treated mice when compared to the controls. Discussion and conclusion: These results suggest that the intake of D. esculentum, even after cooking, may induce infertility by altering the male reproductive function, and therefore, should be evaluated further as a potential antifertility agent.

Improvement of sexual behavior and semen quality by therapeutic approach and zinc supplementation on Karan Fries

Sub-fertility is one of the major concerns in crossbred bulls as 42.98% bulls are getting culled due to sub- fertility problems therefore, a study was planned on 18 Karan Fries bulls producing poor quality semen or not mounted. They were randomly grouped into 3 treatment groups of 6 animals each and 2 normal bulls were kept as control. Sexual behavior and semen quality were studied during pre-therapy (1 month), therapy (90 days) and post- therapy (90 days) period. Data were analyzed using ANOVA. During therapy period, first group treated with 10 g of herbal product orally, second group with 0.0082 mg of GnRH intramuscularly at 10 days interval and third group with 80 ppm of zinc for 90 days for each group. The results depicted that herbal and GnRH treatment significantly reduced the reaction time, whereas herbal, GnRH treatment and zinc supplementation significantly improved the libido score and reduced per cent of non-mounts during the therapy and post-therapy period. The volume of semen and level of testosterone improved in all treatments groups. Herbal, GnRH treatment and zinc supplementation increased the average number of good and medium quality ejaculates/month/bull and maximum efficiency achieved during the treatment period. There was increase in percent live sperm and decrease in abnormality in herbal treated and zinc supplemented group. Therefore, it can be concluded that GnRH therapy, herbal treatment and zinc supplementation will be effective to overcome the sub-fertility problem in crossbred bulls.

Addition of antioxidants improves quality of ram spermatozoa during preservation at 4 °C

The aim of this study was to investigate the effects of different antioxidants on the quality of ram spermatozoa. For this purpose, ejaculates were collected from six adult healthy rams and extended with Tris egg yolk fructose extender without (Control) or with different antioxidants like ascorbic acid (4.5mg/ml), butylated hydroxytoluene (BHT, 1mM) and taurine (25mM) and stored up to 72h at 4°C. The quality of spermatozoa on the basis of percentage of sperm motility, live sperm, morphological abnormalities, intact acrosome and hypoosmotic swelling test (HOST) reacted spermatozoa was evaluated at 0, 24, 48 and 72h of preservation at 4°C. The percent sperm motility was significantly (P<0.01) higher in BHT and taurine and significantly (P<0.01) lower in ascorbic acid than the control at 72h of preservation. The percent live sperm did not differ significantly (P>0.05) amongst the extenders containing different antioxidants and the control. The morphological abnormalities were significantly (P<0.01) lower in BHT and taurine than ascorbic acid and control during preservation up to 72h at 4°C. The percent HOST reacted spermatozoa was significantly (P<0.01) higher for BHT and significantly lower (P<0.01) for ascorbic acid than control. The percent intact acrosome was significantly (P<0.01) higher in all the extenders containing antioxidants than control. In conclusion, BHT and Taurine were found to improve the quality of ejaculated ram spermatozoa during preservation for 72h at 4°C.

Effect of dietary fish oil supplementation on ram semen freeze ability and fertility using soybean lecithin– and egg yolk–based extenders

Ram semen cryopreservation is not efficient for artificial insemination in commercial herds. Beneficial effects of dietary fish oil have been evaluated for cryopreservation of ram semen in soybean lecithin (SL) and egg yolk (EY)–based extenders. A factorial study (two diets × two extenders) was used to analyze the effects of two diets supplemented with fish oil (n-3 fatty acid) or palm oil (saturated fatty acids; [SFAs]) to freeze ram semen in two extenders containing SL or EY. Motility characteristics, membrane integrity, abnormal morphology, mitochondria activity, acrosome integrity, apoptotic status, and fertilizing ability were assessed after freeze-thawing. Although diet had significant (P ≤ 0.05) effects on the quality parameters of frozen-thawed sperm, effects of extenders on these traits were not significant (P > 0.05). The higher significant (P ≤ 0.05) percentage of total motility and progressive motility were observed in n-3/SL (44.83 ± 1.56 and 28.33 ± 1.4) and n-3/EY (43.33 ± 1.56 and 28.50 ± 1.4) than SFA/SL (32.16 ± 1.56 and 14.00 ± 1.4) and SFA/EY (31.66 ± 1.56 and 12.66 ± 1.4) groups. Moreover, n-3/SL and n-3/EY produced the higher significant (P ≤ 0.05) percentage of membrane integrity of sperm (39.83 ± 1.4 and 37.33 ± 1.4) than SFA/SL and SFA/EY (29.83 ± 1.4 and 28.5 ± 1.4). For viability results, the higher significant percentage of live sperm was observed in n-3/SL and n-3/EY (43.16 ± 1.38 and 45.66 ± 1.38) than SFA/SL and SFA/EY (28.66 ± 1.38 and 27.5 ± 1.38). For fertility trials, n-3–based diets (n-3/SL and n-3/EY) improved significantly (P ≤ 0.05) pregnancy rate (44% and 46%), parturition rate (42% and 42%), and lambing rate (46% and 44%) compared with the SFA-based diets (SFA/SL and SFA/EY). No interaction effects have been found between diets and extenders (P > 0.05). It seems that dietary fish oil can improve the semen performance after freezing–thawing process and artificial insemination aside from type of extenders.

Fertility and flow cytometry study of frozen-thawed sperm in cryopreservation medium supplemented with soybean lecithin

Semen cryopreservation can provide genetic resources for a large number of females from a small number of superior males. Optimization of cryopreservation media to achieve the highest quality of post-thaw semen is crucial. Soybean lecithin has evaluated as a plant-based cryoprotectant for substitution of egg yolk in ram semen extender. Flow cytometric and fertility assessments were applied following cryopreservation procedure in two experimental groups (SL group: extender containing 1% w/v soybean lecithin and EY group: extender containing 20% v/v egg yolk). The higher percentage of live sperm and the lower percentage of dead sperm were obtained in SL (47.66 ± 1.38, 52.33 ± 1.69, respectively) extender compared to EY (41.16 ± 1.38, 58.83 ± 1.69). For motion characteristics, plasma membrane integrity, acrosome integrity and mitochondria activity, no significant difference was observed between SL and EY extenders. In artificial insemination experiment, there was no significant difference in pregnancy rate, lambing rate and twining rate between SL and EY extenders. It can be concluded that SL extender can be an efficient alternative extender to preserve ram sperm during cryopreservation procedure without adverse effects.

KUALITAS SPERMATOZOA ASAL CAPUT, CORPUS, DAN CAUDA EPIDIDIMIS PADA KERBAU RAWA (Bubalus bubalis carabenensis)

The purpose of this research was evaluate the quality of spermatozoa concentration in the caput, corpus and cauda of the swamp buffalo epididymis (Bubalus bubalis carabanensis). Method of this research was to exploration to 13 epididymides of eight swamp buffaloes were obtained from Banjar and Banjarmasin slaughterhouses,evaluated the quality of spermatozoa in caput, corpus and cauda of epididymis. Quality of collected-spermatozoa including spermatozoa motility, percentage of live spermatozoa, spermatozoa concentration and percentage of abnormality. Result of this study showed that mean of each of caput spermatozoa motility, percentage of live spermatozoa, spermatozoa concentration and percentage of abnormality; 0%, 45.43% (31.87–72%), 189,62 x106 (40–480 x106) and 56.16 %(44.34–66.53%), corpus ;2.77% (1–9%), 58.73% (45.14 –76%), 152.31 x106 (45 – 345x106), and 47.61 %(23.92 – 60.15%), cauda;53.46% (20 – 70%), 74.32 % (56.68 – 83%), 1,459.62 x106 (825 – 2,340x106), and 34.60%(15.89 –50.04%). In conclusion, spermatozoaofcaudaepididymis could be used in artificial insemination program.Keywords: Spermatozoa, epididymis, swamp buffalo.

NT-proCNP as a new indicator of asthenozoospermia

PurposeC-type natriuretic peptide (CNP) is a proinflammatory peptide. The highest concentration of CNP is found in male reproductive organs. This study aims to analyze the role of N-terminal C-type natriuretic propeptide (NT-proCNP) as a new indicator of asthenozoospermia. Material/methodsSemen was collected after 3–5 days of sexual abstinence from 86 men. The participants were between 25 and 38 years old, 51 of which had asthenozoospermia and 35 of which had normal sperm motility. Semen was analyzed for the concentrations of NT-proCNP, spermatozoa, percentages of live sperm, and sperm exhibiting a specific type of movement. The laboratory tests and analyses were performed using accepted methods and under appropriate conditions. ResultsA significant difference in the concentration and motility of spermatozoa was observed between the focus and control groups. The concentration of spermatozoa in the focus group was significantly lower than in the control group (median: 38.5 vs. 69.8mln/ml [p=0.016] respectively). The progressive motility of spermatozoa demonstrated a significantly lower performance in the focus group than in the control group (median: 10.4% vs. 45% respectively). The concentration of NT-proCNP was significantly higher in the focus group (median: 29.1 vs. 17.9pmol/l; p<0.001). The Area under the Receiver Operating Characteristic (AUROC) curve for the concentration of NT-proCNP – as an indicator of asthenozoospermia – was 0.733. Participants with a concentration of NT-proCNP higher than 28.8pmol/l had asthenozoospermia with 52.9% sensitivity and 94.3% specificity. ConclusionsNT-proCNP – an indicator of inflammatory reaction – should be evaluated as an indicator of asthenozoospermia.

Effect of LDL Addition Into Selected Bull Sperm Diluters on Resistance of Spermatozoa Against Cold Shock

The aim of work was to determine the effect of LDL cholesterol addition into selected diluters on the resistance of spermatozoa against cold shock and on their short-term survivability during cold test. The hypothesis was that the addition of LDL cholesterol will positively affects sperm resistance to cold shock and ensures a higher survivability of spermatozoa during short-term cold survival test. Four bulls of different breeds and ages, from the same sire insemination center were used. A total of eight semen collections were processed. Each ejaculate was divided into 6 portions (3 controls and 3 samples). Three commercially produced diluters, AndroMed®, Bioxcell®, and Triladyl® were used, each in standard and LDL enriched variants. In the case of AndroMed® or Bioxcell®, 6% of LDL was simply added. In Triladyl®, 10% of LDL replaced the standard egg yolk component. Spermatozoa resistance to cold shock was evaluated by the percentage of live sperm using Eosin-Nigrosine staining. The results showed the influence of bull individuality as an important factor. It is possible to recommend Bioxcell® with addition of LDL cholesterol in 6% concentration, which survivability was 69.17% at the beginning of the test, and 52.94% after 2 hours of incubation.

Effect of washing on the post-thaw quality of cryopreserved ram epididymal spermatozoa

Aim:The aim of the study was to evaluate the effect of washing on the post-thaw quality of ram cauda epididymal spermatozoa (P1: Unwashed, P2: Washed).Materials and Methods:Fresh testicles of adult healthy slaughtered rams were collected and transported to the laboratory in an ice chest, where they were weighed, and cauda epididymides were separated. These cauda epididymides were used for recovery of spermatozoa in tris-citric acid fructose buffer by incision method. Spermatozoa samples showing ≥70% progressive motility were pooled and processed further. The mean values (±standard error) of various parameters such as the percentage of sperm motility, live sperm, intact acrosome, and hypo-osmotic swelling test (HOST) reacted spermatozoa were recorded.Results:In this experiment, the percent sperm motility, live spermatozoa, and intact acrosome both at pre-freeze and post-thaw were higher (p>0.05) in P1 than P2. However, the post-thaw percent HOST reacted spermatozoa was slightly higher (p>0.05) for P2 than P1.Conclusion:Washing of cauda epididymal spermatozoa has no significant adverse effect on the quality during cryopreservation. Therefore, this processing method can be applied wherever necessary before the extension of the recovered spermatozoa sample in different ram extenders.

Evaluation of Cauda Epididymal Semen Quality of Crossbred Bulls in the Tropics

The objective of the study was to evaluate the quality of semen retrieved from cauda epididymis of crossbred bulls in the tropics. Testes from one hundred and twenty six recently slaughtered cross bred bulls in the tropics were used for the analyais. The total and progressive motility percentage obtained for epididymal semen were 49.17±9.26 per cent and 27.5±9.11 per cent respectively. The mean concentration obtained for epididymal semen was 37,175x106 ±7612x106 per ml. The mean percentage of live and dead sperms was 84.5±8.02 per cent and 15.5±8.02 per cent respectively. The mean percentage of normal spermatozoa, spermatozoa with abnormal heads, abnormal tails, spermatozoa with a proximal protoplasmic droplet and distal protoplasmic droplet were 35.67±2.30, 3.17±1.58, 2.33±0.61, 11.67±4.01 and 47.17±3.17 per cent respectively. Int J Appl Sci Biotechnol, Vol 4(1): 130-132

Metabolic activity of sperm cells: correlation with sperm cell concentration, viability and motility in the rabbit

The resazurin reduction test (RRT) is a useful technique to assess the metabolic rate of sperm cells. RRT depends on the ability of metabolically active cells to reduce the non-fluorescent dye resazurin to the fluorescent resorufin. The aim of this study was to develop a vital fluorometric method to evaluate metabolic activity of rabbit sperm cells. Twenty-five rabbit males were included in the study. Viability and morphology, motility and metabolic activity were evaluated using an eosin-nigrosin staining, a computer-assisted semen analysis (CASA) and the RRT, respectively. Spearman rank correlation analysis was used to determine the correlation between RRT and semen parameters. After evaluation, a concentration of 10 × 106 sperm cells/ml was selected for further experiments with RRT. No significant correlation was found between the RRT results and the motility parameters. However, after RRT a significant positive correlation between relative fluorescence units and the percentage of alive spermatozoa (r = 0.62; P = 0.001) and a negative one with the percentage of sperm cells with acrosomic abnormalities (r = -0.45; P < 0.05) were detected. The vital assessment of metabolic rate of sperm cells by RRT could provide more information about semen quality than other routine semen analysis, correlating with sperm viability and acrosome status information.

The characterisation and cryopreservation of Venda chicken semen

Abstract Objective To characterize Venda cocks semen, find a suitable short-term diluted semen storage temperature, find a suitable cryopreservation cryoprotectant and to investigate cryoprotectant toxicity. Methods Semen was collected from six Venda cocks and evaluated macroscopically for semen volume, pH and sperm concentration. Microscopic sperm characteristics examined included were total motility (rapid, medium and slow) progressive and non-progressive motility. Velocity characteristics included curvilinear and straight-line velocity, average path velocity, linearity, straightness, wobble, amplitude of lateral head displacement and beat cross frequency. Results Results showed that the average semen volume was 0.3 ± 0.1 mL, the pH 6.9 ± 0.4 and the sperm concentration (6.8 ± 79.8) × 109/mL. A positive correlation was observed between body weight and semen volume (r = 0.38). Similarly a significant difference between the initial sperm total motility (TM%) of 87.5 ± 8.6 and samples stored for 24 h at 5 °C (55.0 ± 8.0) and 25 °C (30.6 ± 6.1) was recorded. The percentage live and normal sperm was 87.0% and 93.5% (P Conclusions In conclusion, the Venda cock semen was subsequently found to have a higher TM% when stored in vitro at 5 °C. DMSO and EG were found to be suitable for the cryopreservation of Venda cock semen.

Practical use of percoll density gradient centrifugation on sperm sex determination in commercial dairy farm in Thailand

Our experiment demonstrated an extension of Percoll® Density Gradient Centrifugation to use as a selection tool for sperm sexing developed for dairy farmer. We develop a simple protocol for bovine sperm sexing and used for artificial insemination (AI) and in vitro fertilization (IVF) for embryo transfer (ET). The quality of sperm after centrifugation in each layer were significantly different (P &lt;0.01). There was no significant difference between percentage of live sperm after centrifugation and fresh sperm before centrifugation (P &gt;0.05). The phenotype and swimming capacity of sperms in each layer were significantly different (P &lt; 0.01). It was found that the most appropriate sperm in the layer after centrifugation and after quinacrine staining was between 65-70% Percoll®. X-bearing spermatozoa at this layer was 60.75% and motility was 95.86±0.46%. After insemination, the pregnancy ratio in the experiment was 40% which is similar to the percentage of normal AI. The number of female offspring in the experiment was 71.4% in selected sperm vs 50% in non-selected sperm AI. In the In Vitro fertilization experiment, sperm from our 7 layer Percoll® gradient concentration can fertilize and produce blastocyst embryo successfully. The simple 7 layer Percoll® gradient concentration protocol is appropriate for dairy farmer to do as a pre-AI step to increase their female offspring in their herd.

Boar and season effects on some parameters of semen fertilizing potential

In order to determine the more accurate fertile potential of sperm, it seems that the conventional parameters of boar semen quality (the ejaculate volume, sperm concentra?tion, progressive motility, percentage of live sperm and of those with intact acrosomal mor?phology) are insufficient. Since recently, there have been numerous studies proving that protein concentration in sperm plasma has high positive correlation with boar fertile po?tential. The research objective was to determine the effect of boars as well as the season on the variation of protein content in the sperm plasma. For the research there were used spermal fractions of 2 boars with high (V-boar) and 2 boars with low (N-boar) protein con?tent in spermal plasma. The ejaculates of boars were taken once a week, for a month, during one year (4 ? 12 = 48 ejaculates per boar). For protein analysis in the spermal plas?ma, the samples were prepared by centrifugation. The ejaculate volume, protein concen?tration and progressive motility varied considerably (p &lt; 0.05 or p &lt; 0.01) among the boars as well as in one and the same boar. The variations of the same parameters were deter?mined also during both warm and cold season. On the other hand, protein concentration was rather constant, and in V-boars (ranged from 4 to 4.5%) while in N-boars (ranged from 2.3 to 2.6%). The season did not significantly affect (p &gt; 0.01) protein content in sperm plasma (V-boars: 4 to 4.5% in warm and cold season; N-boars: 2.3 do 2.6% in warm and 2.3 to 2.5% in cold season). The obtained results showed that measurement of protein con?tent in boar sperm plasma could be a useful method for their ranking, based on fertile po?tential of fresh semen.