Tris-egg Yolk Research Articles

Fertilitas Semen Beku dalam Tris Kuning Telur dan Skim yang Diberi Omega-3 pada Sapi Simmental dengan Ransum Berimbuhan Seng dan Selenium Minimal

This study aims to examine the quality of frozen semen in Tris egg yolk (TEY) extender and skimmed milk extender with or witout omega-3. A total of 18 Simmental bulls belong to Lembang Artificial Insemination Centre were divided into three groups. Each was fed with standard feed (R1), standard feeds supplemented with minimal Se and Zn (R2) and standard feed supplemented with maximal of Se and Zn concentration. Semen were collected using an artificial vagina and were evaluated macro- and microscopically. The semen then were divided into four tubes and each diluted with Skimmed, SkimmedOmega 3, TEY or TEY-O. The semen was then packed into a 0.25 ml straw and equilibrated at 5 oC for 4 hours, then frozen above liquid nitrogen vapor, and stored in liquid nitrogen container (-196 oC). The qualities of frozen semen were evaluated on the motility, individual score, viability and integrity of the plasma membrane of sperms. Sperm motility of bulls fed with standard feed (R1) in TEY extender and R3 in TEY and TEY-Omega-3 extender were higher (p <0.05) compared to the other combinations. No difference was found on the individual score. The viability of sperms in bulls fed with standard feed in SkimmedOmega-3 extender was higher than the other treatments and the highest sperm plasma membrane integrity was demonstrated by sperm in bull feeding with R2 in TEY extender.

Kualitas Semen Segar Sapi Bali Umur Produktif dan Non-produktif serta Penentuan Konsentrasi Krioprotektan dalam Pengencer Tris Kuning Telur

The aim of this study was to compare the quality of Bali bull fresh semen in productive group (below 10 years old) with over productive group and to evaluate the frozen semen quality of bali bull after cryopreservation in Tris egg yolk extender using Dimethyl formamide (DMF) and glycerol in three different concentrations (5%, 6%, and 7%). Semen was collected using artificial vagina. Semen then evaluated macro and microscopically. Macroscopic evaluation conducted visually and microscopic evaluation with the aid of Computer Assisted Sperm Analysis CASA. Immediatelly after evaluation, semen were divided into six aliquots and diluted with Tris egg yolk (TEY) glycerol 5% (TEYG5), TEY-glycerol 6% (TEYG6), TEY glycerol 7% (TEYG7), TEY-DMF 5% (TKTD5), TEY-DMF 6% (TEYD6) and TEY-DMF 7% (TKTD7). Results of experiment showed that there was no difference (P>0.05) among the all parameters, except for semen volume. Semen volume of productive group was higher than over productive age group. No difference was found in post thawing motility of the spermatozoa in TEY extender with 5% glycerol, and TEY with 5%, 6% and 7% DMF. The higher recovery rate of bali bull spermatozoa demonstrated by spermatozoa in TEY extender with glycerol 5% and DMF 6% (63.33±2.40 dan 68.67±2.33). It was concluded that bali bull semen quality did not differ between productive and over productive ages and post thawing semen quality of bali bull in TEY with 5-7% DMF or 5% glycerol was not also different.

Comparison of sperm adenosine triphosphate content, motility and fertility of immobilized and conventionally cryopreserved Norwegian Red bull semen

Estrus detection and timing of AI remains a challenge in cattle breeding. Prolonging spermatozoa lifespan after AI, making sperm cells available over an extended period, could make timing of AI relative to ovulation less crucial and improve fertility. Immobilization of sperm cells by the patented SpermVital technology in an alginate gel will provide a gradual release of spermatozoa after AI. The first aim of this study was to examine fertility, measured as non-return rate after 56 days (NR56), of SpermVital (SV) processed semen with reduced sperm cell number per dose compared to earlier studies, and compare with conventionally processed semen in Biladyl, a proprietary version of the egg yolk Tris semen extender. The second aim was to examine in vitro sperm quality post-thaw and after thermal stress. The third aim was to examine potential correlations between in vitro sperm parameters and NR56. Ejaculates from 16 Norwegian Red young bulls were split in three, processed and cryopreserved as Biladyl semen (B15; 15 million spermatozoa/dose) or by SpermVital technology (SV25; 25 million spermatozoa/dose or SV15; 15 million spermatozoa/dose). 1400 semen doses were produced per bull and distributed throughout Norway for a blinded field trial. Fertility was recorded as NR56 after first AI (N = 7155). Two ejaculates from each bull were randomly selected for in vitro experiments. B15 and SV15 semen samples were analyzed for motility by computer-assisted sperm analysis, viability and acrosome integrity by flow cytometry and ATP content by bioluminescence assay, post-thaw and after thermal stress. The AI trial detected no differences in NR56; least square means being 75.5% (B15), 75.6% (SV25) and 74.8% (SV15) (p > 0.05). There were no differences in total motility and progressive motility post-thaw, however, after three hours incubation at 38 °C, SV sperm motility and progressivity were higher for SV15 than for B15 spermatozoa (p < 0.05). The percentage of acrosome intact live sperm cells was higher for SV15 than B15 spermatozoa at all timepoints analyzed (0 h, 3 h, 24 h, p < 0.05). B15 semen showed a higher ATP level than SV15 at 0 h (p < 0.05), while SV15 sperm cells had higher ATP levels after 3 and 24 h (p < 0.05). No association was detected between in vitro sperm parameters and NR56. In conclusion, SV15, SV25 and B15 semen yielded equal fertility after AI. However, there were differences in sperm quality, as SV15 spermatozoa displayed higher motility, viability and ATP levels after thermal stress than B15 spermatozoa (p < 0.05).

Additional Freeze Drying Fig Fruit (Ficus Carica L) Filtrate into Tris Egg Yolk Extender and Its Effect on Sperm Membrane Integrity and Acrosome of Kacang Buck

A study was designed to determine Kacang buck sperm membrane integrity and acrosome reaction as to the effect of different concentration of freeze drying fig fruit filtrate in tris egg yolk based extender. This study used 5 proven fertility Kacang goats aged 3-4 years, maintained by Faculty of Animal Sciences, Mataram University, Indonesia. Semen was collected by artificial vagina at every five days. The collected semen was divided into four aliquots in accordance to the treatments extender such as Control (0 gr), T1 (0.02 gr), T2 (0.04 gr) and T3 (0.06 gr) freeze drying fig fruit filtrate in tris egg yolk based extender (gr/v), respectively. Plasma membrane integrity and intact acrosome after re-concentration and preserved at 5ºC were assessed visually at 0 and every 24 hours for 5 consecutive days. The statistical significance of the result was evaluated by a one way ANOVA for completely randomized design analysis of variance. Data were presented as Mean±SD. Results suggest that the mean percentages of sperm membrane integrity in T0, T1,T2 and T3 at 96 h post extended and preserve at 5ºC were 34.3±5.3, 40.6±4.7, 44.8±5.4 and 42.1±5,1, respectively. The mean percentages of sperm acrosome intact were 16.4±4.8, 18.5±1.9, 21.6±3.1 and 19.6±2.8, respectively. The results of the study suggested that additional 0.04gr freeze drying fig fruit filtrate into tris egg yolk based extender have a significant preservation effect on both spermatozoa membrane integrity and acrosome intact of kacang buck.

Supplementation of Nigella sativa Oil and Honey Prolong the Survival Rate of Fresh and Post-Thawed Goat Sperms

Some chemical compositions of polyphenol antioxidants found in Nigella sativa and honey have potential to protect sperm membrane from lipid peroxidation. Therefore, we study the effect of in vitro supplementation of Nigella sativa oil and honey on sperm parameters; motility, progressive score, and abnormalities in fresh and post-thawed goat semen. Pooled semen from five sexually matured males Jermasia goats between aged 2 to 3 years old, with average live weight of 35.9±5.7 kg and average body condition score (BCS) of 1.7±0.45 were used in this study. The semen were added with tris-egg yolk extender and combination of supplements (Nigella sativa oil or/and honey), and divided into groups; Control (no supplementation), Treatment 1 (0.5% v/v Nigella sativa), Treatment 2 (2% v/v honey), and Treatment 3 (0.5% v/v Nigella sativa and 2% v/v honey). All parameters of fresh semen were immediately assessed after semen collection and parameter of post-thawed semen were assessed after 48 hours (2 days) of semen cryopreservation. The fresh sperm parameters were observed at 0, 0.5, 1, 1.5, and 2 hours after semen collection. Results showed that the motility of fresh semen at 1.5 and 2.0 hours after collection treated with Nigella sativa oil (Group 1; 73.8% and 72.0% respectively) and honey (Group 2; 73.3% and 72.0%; respectively) were 20% (1.5 h) and 8% (2.0 h) lower than Controls (93.0% and 79.8% of motility) (P<0.05). The progressive score and sperm abnormality were not significant between groups. While, the motility of post-thawed semen was higher (P<0.05) in treatment 3 (60.33±10.08 of motility; supplemented with Nigella sativa and honey) compare to Controls (24.33±8.17 of motility) at 0 and 0.5 hour after thawing. Thus, we suggested that Nigella sativa oil and honey show a potential as a supplement to goat’s extender with the ability to protect sperm membrane from damage due to oxidative stress and could prevent ice crystal formation during cryopreservation.

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Anti-apoptotic effects of minocycline on ram epididymal spermatozoa exposed to oxidative stress

Cryopreservation causes major damage to sperm cells and thereby, increased apoptosis has been documented as physiologically programmed cells death. The goal of current study was to evaluate the anti-apoptotic effects of minocycline compared to sericin added to diluents on frozen-thawed spermatozoa. Epididymal spermatozoa isolated from 50 pairs testes with motility >80% and total morphological abnormalities <10% were pooled, divided into 9 aliquots and used for cryopreservation. The semen samples were re-suspended with basic Tris egg yolk diluent containing minocycline (5 and 10 mg) and tert-butyl hydro peroxide (10 and 20 μm) and sericin (0.5 gr). Total sperm motility and progressive motility were evaluated by Computed Assisted System Analysis (CASA). Acrosome and plasma integrity, viability, hypo osmotic swelling test, malondialdehyde concentration, DNA fragmentation test, mitochondrial activity, apoptotic features, caspase activity and level of H2O2 and O2 were assessed as completed characteristics of frozen-thawed spermatozoa. The results indicate that 10 mg minocycline added to extender resulted in significant (P < 0.05) enhancement of total motility, progressive motility and viability of post-thawing spermatozoa. The results of plasma membrane demonstrate significantly (P < 0.05) higher value of both minocycline concentrations with 10 μm tert-butyl hydro peroxide. In regard to caspase activity, diluents supplemented with 5 and 10 mg minocycline improved significantly (P < 0.05) the rate of viable spermatozoa. The results of malondialdehyde concentration show diluents supplemented with 10 mg minocycline and 0.5 gr sericin were significantly (P < 0.05) lower than other extenders. The ratio of sperm chromatin dispersion stained spermatozoa illustrated the higher rate in extenders containing 10 mg minocycline plus 10 μm tert-butyl hydro peroxide, 5 mg minocycline plus 10 μm tert-butyl hydro peroxide and 5 and 10 mg minocycline were significantly (P < 0.05) than other treatments. Moreover, the results of mitochondrial activity show significantly (P < 0.05) an improvement by supplementation of 5 and 10 mg minocycline. Regarding the apoptosis occurrence and intracellular ROS, the results demonstrate significantly (P < 0.05) higher live post thawed spermatozoa and lowest value for diluents containing 10 and 5 mg, respectively. In conclusion, the results indicate that addition of 10 mg minocycline seems to reduce the apoptotic marker during cryopreservation of epididymal ram spermatozoa.

Effect of Propolis Ethanolic Extract Supplementation to Ram Semen Extenders on Sperm Characteristics, Lipid Peroxidation and some Enzymatic Activities in Seminal Plasma in Chilled Semen

This study designed to examine effects of adding different concentrations of propolis ethanolic extract (PEE) as antioxidant to ram semen extenders; tris egg yolk (TEY) or tris-soybean lecithin (TSBL) on sperm characteristics, lipid peroxidation and enzymatic activities in seminal plasma of ram semen preserved at 5°C for 48 hours. Semen was collected from five adult Rahmani rams using an artificial vagina twice/week for 5 weeks. Pooled semen was extended with TEY or TSBL supplemented with different concentrations from PEE (0, 0.1, 0.5 and 1.0 mg/ml). Polyphenols and flavonoid compounds in propolis ethanolic extract were identified. In each treatment, percentage of progressive motility, livability, and abnormality were determined. Also, percentages of sperm membrane integrity or chromatin damage were determined. Concentrations of total antioxidant (TAO) and malondialdehyde (MDA), and activity of acid phosphatase (ACP), lactate dehydrogenase (LDH), and asprtate (AST) and alanine (ALT) transaminase, were determined. Results showed that the most effective polyphenolic compounds in the prolopolis ethanolic extract was the chlorogenic acids, catechein, protocatchuic and pyrogallol (120.623, 53.006, 23.907 and 11.048 mg/100g extract, respectively). Luteo.6-arbinose 8-glucose was the predominant identified flavonoid component in propolis extract (5268.78ppm), followed by acacetin, hisperidin, apig.6-rhamnose8- glucose, apig.6-glucose8-rhamnose, kamp.3-(2-p-comaroyl) glucose and luteo.6-glucose 8-arbinose with concentrations )647.53, 561.09, 366.47, 150.46, 144.51 and 109.42 ppm, respectively. The effect of propolis ethanolic extract supplementation to ram semen extenders was significant (P<0.05) only on the overall percentages of sperm progressive motility and sperm chromatin damage while, sperm livability, abnormality and membrane integrity were not affected significantly (P<0.05). Concentration of TAC was numerically (P= 0.82) higher in TEY extender with different concentrations of PEE than TSBL. However, MDA was lower (P<0.05) in free TEY, followed by TSBL with PEE. Activity of LDH, AST and ALT tended to be lower in seminal plasma of semen extended with TSBL than TEY. These results showed beneficial effects of soybean lecithin and propolis ethanolic extract in tris based extender on progressive motility and chromatin integrity of spermatozoa up to 48 h of preservation at 5°C. Also, lipid peroxidation and enzyme activity were lower in TSBL than TEY. In addition, soybean lecithin can effectively replace egg yolk as a protective additive with propolis ethanolic extract for preservation extender without any detrimental effects on post-chilling semen quality in rams.

Effect of reduced glutathione supplementation on cryopreservation induced sperm cryoinjuries in Murrah bull semen

The experiment was conducted to study cryopreservation induced sperm cryoinjuries and the protective effect of reduced Glutathione supplementation in Murrah bull semen. A total of 20 semen ejaculates were split into two parts after initial examination and were extended in glycerolated egg yolk TRIS diluter (Control group) and glycerolated egg yolk TRIS diluter + 0.5 mM reduced Glutathione (Treatment Group). The diluted semen samples were loaded into 0.25 ml French mini straw and sealing of straws were done. Thereafter, semen straws were kept for equilibration for 4 h at 5 °C and semen was frozen using a standard cryopreservation protocol in automatic biological freezer. Post-thaw analysis was performed after 24 h of semen storage in liquid nitrogen. Fresh and post-thaw sperm assessments included sperm motility, viability (SYBR-14/PI assay), mitochondrial function (JC-1/PI assay) and plasma membrane integrity (HOST). Cryopreservation of semen in liquid nitrogen induced a marked reduction in post-thaw sperm motility, viability, mitochondrial function and plasma membrane integrity and increased moribund type of sperm (SYBR-14/PI assay) in control group as compared to reduced glutathione treated group. There were significant (P < 0.05) cryo injuries in frozen-thawed spermatozoa following cryopreservation in buffalo bull semen. The supplementation of reduced glutathione in treatment group exhibited significantly (P < 0.05) lower cryoinjuries during cryopreservation and semen storage in liquid nitrogen. From the study it was concluded that, spermatozoa from Murrah bulls are susceptible to injuries due to cryopreservation in liquid nitrogen, but these cryo induced damage can be protected significantly (P < 0.05) by the use of reduced Glutathione.

Cryoprotective role of organic Zn and Cu supplementation in goats (Capra hircus) diet

The current study focused on cryopreservation and assessment of characters of post-thaw semen of indigenous Osmanabadi bucks maintained with standard diet, supplemented with different concentrations of organic zinc (Zn), copper (Cu) or in combination, for a period of 180 days. The different doses of organic Zn and Cu were fed per kg DM basis, Zn groups (low: Zn20, medium: Zn40 and high: Zn60), Cu groups: (low: Cu12.5, medium: Cu25 and high: Cu37.5) and combination of Zn + Cu groups (low: Zn20 + Cu12.5, medium: Zn40 + Cu25 and high: Zn60 + Cu37.5) respectively. The control group bucks were maintained mainly on the basal diet without any additional mineral supplementation. Two hundred and forty (240) semen samples were collected from 40 bucks aged 11 months, through electro ejaculator method, processed and analysed for sperm quality parameters both at pre freeze and post-thaw stage. The semen samples were diluted in Tris egg yolk extender, cooled and equilibrated for 4 h at 5 °C, cryopreserved using programmable freezer (PLANER Kryo 360–1.7) and stored at −196 °C. The organic trace minerals (Zn, Cu and Zn + Cu) protected the spermatozoa against the cryoinjury and maintained higher post-thaw semen parameters except in high Zn group. Additional feeding of organic Cu and Zn to bucks had a protective role and resulted in higher sperm liveability, plasma membrane and acrosome integrities, motility and velocity and reduced oxidative stress in supplemented goats (P < 0.05).

Improved quality of frozen boer goat semen with the addition of sweet orange essential oil on tris yolk and gentamicin extender

This research aimed to determine the extent of frozen semen quality Boer Goat by essential oils of sweet orange peel in tris yolk and gentamicin extender. Research has been conducted at the Laboratory Loka Penelitian Kambing Potong Sei Putih, Deli Serdang, North Sumatra in February 2017. This study used a completely randomized design with 4 treatments and 5 replications. Treatments are 0.25; 0.5; 0.75 and 1% essential oils as additional diluent. The parameters were measured percentage Motility, membrane integrity, acrosome integrity and viability Boer Goat frozen semen. The results showed that the addition of essential oils as diluent semen was significant (P <0.01) in the percentage motility, Viability, membrane integrity and acrosome integrity Boer Goat frozen semen. Motility, membrane integrity, acrosome integrity and viability was significantly higher in all treated groups than the control group. The best results of all treatments In the study was the addition of essential oil as much as 1%.

Semen Quality of Post-Thawed Local Ram’s in Tris-Egg Yolk Extender with Different Glutathione Level

The aims of this research were to find out the effect of glutathione level on semen quality of local ram and to find out the glutathione level that provide the best quality of local ram semen. This research use Completely Randomized Design (CRD) with five treatments of glutathione level (0 mM, 4 mM, 5 mM, 6 mM and 7mM) and was repeated for 10 times. Data were analyzed using analysis of varians (ANOVA) and differences between treatment was analyzed using Duncan test. The parameter were semen quality consist of motility, intact plasma membrane (IPM), abnormality, and recovery rate. Result of this research showed that glutathione level significantly (p<0.05) affect on motility, IPM, abnormality and recovery rate. Duncan test result showed that recovery rate from treatment of 5 mM glutathione level have significantly higher than other treatment, whereas for motility, IPM and abnormality were not significant different with 4 mM, and 6 mM. It is concluded that semen quality of local ram was affected by glutathione level, and the level of 5 mM in egg yolk tris extender provide the best quality of local ram semen.

Effect of Varying Osmolarity of Tris Extender on Seminal Attributes of Buffalo during Refrigeration

The aim of the present study was designed to investigate the effect of varying osmolarity of tris egg yolk citrate glycerol extender on seminal attributes of buffalo at various hours of refrigeration at 5 oC. Twenty four ejaculates having mass motility ≥ 3+ from 4 bulls (6 ejaculate from each bull) were collected. Each ejaculate was divided into four group’s viz., Group I, Group II, Group III and Group IV, diluted with extenders with osmolarities of 240, 260, 280 and 300, milliosmol/kg, respectively, upto 80×106 sperm/ml. After, dilution, semen samples were filled in French mini straws and kept at 5°C and evaluated at 0, 24, 48, and 72, hours for various seminal attributes such as individual motility, viability, acrosomal integrity, and hypo-osmotic swelling test. No significant difference in individual motility at 0 hour was observed among all the groups. However, at 24, 48 and 72 hours, the individual motility was significantly (P

Alpha-zearalenol negatively influences ram sperm parameters during liquid storage.

This study was aimed to investigate the effects of 17 𝛽-estradiol (E2) and 𝛼-zearalenol (α-ZOL) on motility parameters, plasma membrane integrity, levels of produced nitric oxide (NO) and total antioxidant capacity of Ghezel ram sperm during the liquid storage at 4 ˚C, for various periods of time. Semen samples were collected from four rams and diluted with Tris–egg yolk extender and supplemented with E2 (100 µmol) or different concentrations of α-ZOL (100 pmol, 100 nmol and 100 µmol) at a final concentration of 200 × 106 sperm per mL. We failed to show any significant effect of E2 at 100 µmol concentration on ram’s sperm parameters while α-ZOL resulted in a significant decrease of plasma membrane integrity at 100 µmol concentration (55.40% for α-ZOL vs 62.20% for control) after 96 hr incubation. Alpha-ZOL had decreasing effect on sperm motility parameters including curvilinear velocity and average path velocity at 100 µmol concentration after 96 hr storage. Although remarkable reduction of total antioxidant capacity at high concentration of α-ZOL and long incubation time was found, however no significant changes were recorded in NO level during storage time. It was concluded that the detrimental effect of α-ZOL on ram sperm might be attributed to its induced oxidative stress and damage to the plasma membrane.

Effects of apple and orange juices on quality of refrigerated goat semen

This study investigated the effects of apple and orange juices on quality of refrigerated spermatozoa of goat bucks. Semen samples from WAD goat bucks were diluted with Tris-egg yolk extenders each supplemented with apple and orange juices at 0, 2.5, 5, 7.5 and 10/100 ml of diluents. The diluted semen samples were assessed for sperm viability and malondialdehyde (MDA) concentration after in vitro storage for 240 hours at 5oC. The ability to maintain sperm motility was higher in the extenders with 7.5% orange juice followed by 10% apple juice compared to other treatments (P&lt;0.05). The extenders supplemented with 2.5%, 5% and 7.5% apple juice, and 5% orange juice had higher intact acrosome compared to other treatments and the control (P&lt;0.05). The 10% orange juice had higher percentage membrane integrity compared to other treatments. Consistent and reduced (P&lt;0.05) MDA levels were observed in the extenders supplemented with fruit juices and lower MDA was observed in the extenders supplemented with 10% apple juice compared to other treatments and the control (P&lt;0.05). The findings reveal that additions of the fruit juices to semen extenders to maintain the viability of refrigerated spermatozoa were best at concentrations of 10 ml/100 ml of apple juice and 7.5 ml/100 ml of orange juice.

A two-step dilution tris-egg yolk extender containing Equex STM significantly improves sperm cryopreservation in the African wild dog (Lycaon pictus)

Conservation management of endangered African wild dogs (AWD; Lycaon pictus) can benefit greatly from development of sperm freezing and artificial insemination. Previous freezing attempts yielded nearly 0% motile sperm within 2 h of thawing. In this study, two canine freezing protocols were tested: Protocol 1: a one-step dilution in TRIS-20% egg yolk containing 8% glycerol; and Protocol 2: a two-step dilution in TRIS-20% egg yolk containing a final extender concentration of 5% glycerol and 0.5% Equex STM, coupled with a TRIS-citrate-fructose thawing solution. Semen was collected by electroejaculation from n = 24 AWDs, of which eight ejaculates of sufficient quality (four good quality with initial sperm motility of 75.0 ± 4.4% and four poor quality; showing rapid decrease in sperm motility to 3.3 ± 3.3% prior to freezing) were frozen. For good quality samples, motility and sperm motility index persisted for up to 8 h for Protocol 2, and was higher between 2 and 6 h after thawing with a decrease from 4 h of incubation. Motility dropped to nearly 0% after 2 h incubation for Protocol 1. Viability was higher for Protocol 2 throughout the 8 h of incubation, with a decrease after 6 h, compared to 4 h for Protocol 1. Acrosome integrity was higher for Protocol 2 throughout post-thaw incubation, with a decrease after 2 h for both protocols. Protocols did not differ in normal sperm morphology or DNA integrity. Poor quality samples yielded similar results, except for acrosome integrity, which declined for Protocol 2. In conclusion, a two-step dilution in TRIS-egg yolk-glycerol extender containing Equex STM yields significantly improved post-thaw quality and longevity of AWD spermatozoa, making it suitable for sperm banking and artificial insemination initiatives.

Evaluation of Solvent Toxicity of Plant Extract with Antiviral Action in Refrigerated Goat Semen

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods &amp; Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at 7°C and evaluated up until four h after DMSO addition. Individual progressive motility (MIP), sperm vigor (V), percentage of spermatozoa reactive to hypoosmotic test (HO) and morphologically normal (NOR) were evaluated. IPM, vigor and NOR remained within normal standards for the caprine species in all treatments test. Percentage results of spermatozoa reactive to hypoosmotic was higher in Tris yolk with values ranging between 34.66% to 46.33%. Sperm vigor was positively correlated (r = 0.85) with IPM in the MEM diluted pool of sperm. In Tris yolk, vigor and hypoosmotic test correlated moderately (r = 0.63, r = 0.54, respectively) with IPM. Tris yolk medium added DMSO presented the highest percentage of reactivity to hypoosmotic test in all treatments when compared to MEM added DMSO.Discussion: The fact that DMSO is easily homogenized in water, ethylic alcohol and most organic solvents favors its use in diluting natural extracts. These components are a possible source of products that inactivate caprine arthritis encephalitis virus in sperm, which is the key to promoting the safe use of genetic material of infected breeders, in addition to commercial use of germplasm. In this study, there was no interference of DMSO in the analyzed parameters when added in a maximum concentration of 2.5% to MEM and Tris yolk, which is in accordance with standard values for goats. In addition, Tris yolk may promote greater protection to the membrane of sperm cells, which was demonstrated by hypoosmotic test. This medium could be ideal to be used in new methodologies that incorporate DMSO. In conclusion, DMSO added to dilution media Tris yolk and MEM did not interfere with the quality of refrigerated caprine sperm, which maintained viability. These results indicate that this substance did not present harmful effects to the genetic material, promoting the use as solvent of extracts from plant compounds with potential anti-viral effect. The information in this study may aid new research performed in this area.

Impact of Eurycoma longifolia extract on DNA integrity, lipid peroxidation, and functional parameters in chilled and cryopreserved bull sperm

This study aims to assess the effect of Eurycoma longifolia aqueous extract on chilled and cryopreserved quality of bull sperm. Semen samples were obtained from four Simmental–Brangus. Each sample was divided into two fractions: the first fraction was used for chilling the semen, and the second fraction was used for the freezing process. Both fractions were extended with Tris–egg yolk extender supplemented with 0.0, 0.25, 0.5, 1.0, 2.5, 5.0, and 7.5 mg/ml Eurycoma longifolia aqueous extract. The diluted chilled fraction was chilled at 5 °C for 6 days, whereas the frozen–thawed fraction was frozen in liquid nitrogen. Data revealed that 1 mg/ml E. longifolia aqueous extract yielded significantly (p < .05) higher sperm motility, morphology, viability, and sperm membrane integrity compared with the control group and other treated groups in chilled semen evaluation. For cryopreserved sperm, a significant difference (p < .05) in sperm motility, viability, sperm membrane integrity, DNA integrity, and lipid peroxidation was observed between 5 mg/ml E. longifolia aqueous extract and other treated and control groups. However, no significant difference in the percentage of sperm exhibiting normal sperm morphology was observed among the groups. In conclusion, the addition of 0.25 and 1 mg/ml E. langifolia extract to chilled semen and 5 mg/ml E. longifolia aqueous extract to cryopreserved sperm into Tris–egg yolk extender helps in maintaining superior quality of bull spermatozoa during chilling and freezing.

An endeavor to improve longevity of cryopreserved equine sperm

Exposure of sperm cells to the oxidative stress pending hypothermic storage of semen has been suggested to be responsible, in part, for the decline of their motility and fertility. This study was conducted to evaluate the in-vitro effects of antioxidants (AOs) and / or caffeine on longevity of cryopreserved stallion spermatozoa. Aliquots from the gel-free fraction of semen ejaculates (n=12), collected from 5 Arabian stallions (9-18 years old) of unknown sperm freezability, were mixed 1:1 with a Tris-egg yolk extender (TEYE), centrifuged at 500 χ g for 5 min and sperm cells were frozen in the form of 0.25-ml concentrated pellets after 2-step addition of TEYE supplemented with or without AOs (0.50 mg /ml Na pyruvate, 1 mg / ml Na thiosulfate, 5 mg / ml bovine serum albumin, 0.15 mg / ml zinc chloride and 0.50 mg / ml ferulic acid). The final pre-freeze concentrations of glycerol and sperm cells were 5% and 562-924 χ IO6 / ml, respectively. Frozen pellets from non-AOs and AOs-treated sperm were thawed in a Tris-citric acidglucose solution (40°C) containing 0, 0.49, 0.97 or 1.94 mg / ml caffeine and incubated (140-230 χ IO6 sperm / ml) at 30°C for 3 h. Sperm progressive motility (%) was assessed after centrifugation, before freezing and after 0, 1, 2 and 3 h of thawing. The results revealed significant (P&lt;0.05) effects of sperm treatments only on post-thaw motility. Neither AOs alone nor caffeine alone could significantly ameliorate the maintenance of sperm motility. AOs plus 0.97 or 1.94 mg / ml caffeine were the superior supplements in improving the longevity of stallion spermatozoa.

KUALITAS SEMEN CAIR SAPI SIMENTAL MENGGUNAKAN LARUTAN ISOTONIS KOMERSIAL PADA KONSENTRASI DAN LAMA PENYIMPANAN BERBEDA

Artificial insemination (AI) is a technique to insert the sperm or semen of bull thawed and processed first into the genital tract of female methods and a special tool called insemination gun, efforts to optimize the management of the cement in order to obtain the quality of cement is optimal to do with the selection of diluent cement. One alternative materials cement diluent is an isotonic solution Commercial (LIK). This study aimed to examine the interaction between the concentration and duration of storage of diluent solution isotonic different to the quality of spermatozoa cow simental. This research was conducted at the Regional Technical Implementation Unit of Artificial Insemination Centres (UPTD BIB) Tuah Sakato, Payakumbuh. In April 2016. This study used a randomized block design (RAK) factorial design to Factor A (Concentration LIK) ie A1 = 100 ml Tris Egg Yolk, A2 = (60 ml) Tris Egg Yolk + (40 ml) LIK, A3 = (55 ml) Tris Egg Yolk + (45 ml) LIK, A4 = (50 ml) Tris Egg Yolk + (50 ml) LIK, and factor B (retention) is B1 = 0 day, B2 = 1, B3 = 2 day, days, B4 = 3 days. Each treatment consists of three replicates. Parameters measured were motility, percentage of survival, abrnormalitas, and MPU. These results indicate that an isotonic solution can be used as an alternative Commercial diluent with the best results on the percentage LIK 40 ml, but the effectiveness of the efficiency of storage use was only on the first day (24 hours).