Tris-egg Yolk Research Articles

Effect of Green Tea Extract on Spermatozoa Quality of Peranakan Ongole Bull On Frozen Storage

Low-temperature storage can often result in spermatozoa damage. It occurs due to Reactive Oxygen Species, leading to changes in the lipid composition of the membrane. Antioxidant compounds are needed to prevent ROS. Green tea extract (Camelia sinensis) can be used as an antioxidant agent. This study aimed to determine the effect of green tea extract in tris egg yolk on spermatozoa quality of Ongole Peranakan (PO) cattle. This study used a completely randomized design (CRD) with five extract treatments (0%, 1%, 2%, 3%,4%) and four repetitions. The parameters used were motility, viability, and membrane integrity. The data were analyzed with ANOVA and Duncan's test. Based on the results of the ANOVA test, the addition of green tea extract has a significant effect on spermatozoa quality of PO cattle in frozen storage (P0.005). The best results were obtained by the addition of 3% extract that was able to maintain the quality of post-freezing spermatozoa with average motility, viability, and membrane integrity of 47.44±0.166, 55.14±0.449, and 53.72±0.599. The addition of 3% green tea extract into tris egg yolk can maintain motility, viability, and membrane integrity in spermatozoa of PO cattle inside frozen storage, by adding green tea extract to diluent, spermatozoa can be stored for a long time at low temperatures making it easier for people to carry out artificial insemination.

Short-term preservation of canine sperm-binding ability and other metrics using the INRA-96 in comparison to Tris-egg yolk extender.

The use of assisted reproductive techniques, such as chilled semen, contributes to the maintenance and genetic improvement of canine breeding. The INRA-96 extender is a commercially available, chemically defined medium that was initially developed for the preservation of equine semen and exhibits preservation potential in the canine species. This research aims to evaluate the INRA-96 extender as an alternative for the short-term preservation of canine semen in terms of sperm quality parameters such as motility and kinetic parameters, integrity and functionality of the plasma membrane in fresh and chilled-rewarmed samples, as well as the sperm-binding ability using the perivitelline membrane of the chicken egg as an indicator of the fertilizing capacity of the preserved semen. A total of 18 ejaculates from 9 French bulldogs (two ejaculates per dog) were collected and divided into two aliquots that were diluted in Tris-egg yolk 20% (control) or INRA-96 to a final concentration of 100 × 106 sperm/mL. Samples were refrigerated in a biological incubator at 5°C and evaluated at 0, 24 and 48 h time points. Comparing the two treatments after 48 h of refrigeration, both extenders showed similar values (p < .5) for the majority of kinetic parameters, with the INRA-96 group promoting a total motility of 88.1 ± 2.9%. In addition, the morphology, integrity and functionality of the plasma membrane were preserved above 70% in this group. Dilution with INRA-96 also provided a significantly higher amount of sperm bound (256.2 ± 21.1) to the perivitelline membrane of the egg yolk compared to the sperm-binding rates (p < .05) achieved at the use of Tris-egg yolk (215.2 ± 21 bound spermatozoa) at 48 h. Our study proved similar functional properties of dog sperm cells treated with INRA-96 in comparison to commonly used home-made Tris-based extender during short-time storage.

Ability of Melon (Cucumis melo. L) Fruit Juices Based Tris Diluent on The Quality of Frozen Spermatozoa of Bali Cattle After Thawing

The high potassium content in melon plays a role in maintaining the normal function of the muscles, heart, nervous system, and potassium is also the main regulator of blood pressure. The aim of this study was to evaluate the effect of using melon juice (Cucumis melo L.) as a diluent on the quality of spermatozoa and semen of Bali cattle both before and after freezing. The Banyumulek Artificial Insemination Center produces sperm used in Bali cattle breeding. The technique used was a completely randomized design (CRD) with 4 treatments and 15 replications. The first treatment was 100% egg yolk tris buffer without the addition of melon juice (P0), the second treatment was 80% buffer without the addition of 20% melon juice (P1), the third treatment was 60% buffer with the addition of 40% melon juice (P2). The fourth treatment was 40% buffer with the addition of 60% melon juice (P3). The outcomes showed that the worth of flawless plasma film and mortality upsides of ox-like semen after weakening and subsequent to defrosting were not essentially unique. Be that as it may, not altogether disparate in that frame of mind of motility, level of reasonability, level of ordinariness and post-defrosting anomalies. According to the findings of this study, melon juice can be used as an alternative extender for fresh Bali cattle sperm and preserve the quality of spermatozoa.

The Effect of Alpinia Purpurata K. Schum Rhizome Extract Supplementation in Tris-Egg Yolk Extender on the Quality of Cryopreserved Bali Bull Sperm

The Effect of Alpinia Purpurata K. Schum Rhizome Extract Supplementation in Tris-Egg Yolk Extender on the Quality of Cryopreserved Bali Bull Sperm

Semen Cryopreservation Quality and Sperm Kinematics of Saanen Goats Using Different Diluents

The success of artificial insemination (AI) in small ruminants, especially goats, depends on the quality of frozen semen. Therefore, the current study aimed to determine the quality of various diluents, including tris-egg yolk, AndroMed®, and OviXcell®, on semen quality. The fresh semen samples from three male Saanen goats aged 1.5-2 years were collected and the mean individual motility of samples was recorded at 70%. The cryopreservation quality of the semen was evaluated based on motility, viability, abnormality, and total sperm motility (TSM) indexes. The present laboratory experiment was performed with 3 treatments and 10 repetitions. The treatments in this study were T0 (tris-egg yolk), T1 (AndroMed®), and T2 (OviXcell®). The results showed no significant difference in the parameters of motility, viability, abnormality, and TSM among the treatment group. The kinematic parameters’ average path length, velocity curved linear, and linearity showed a significant difference in all treatment groups. However, there were no significant differences among the three groups in terms of motility, progressiveness, distance curved line, distance straight line, average velocity path, velocity straight line, straightness, amplitude lateral head, beat cross frequency, and wobble kinematic parameters. Motility was higher in T2 than in T0 and T1, viability was higher in T1 than in T0 and T2, and abnormality was lower in T1 than in T0 and T2. In conclusion, the use of various diluents, such as tris-egg yolk, AndroMed®, and OviXcell®, can maintain the quality of frozen spermatozoa for over 24 hours, including motility, viability, abnormality, and TSM. Kinematic parameters obtained using CASA IVOS II can provide relevant information for various parameters using these diluents.

BOER BUCK SPERM QUALITY IN EXTENDER SUPPLEMETED WITH NON-ENZYMATIC ANTIOXIDANTS AND STORAGE AT COLD TEMPERATURE

The aim of this study was to determine the durability of standard extender tris egg yolk supplemented with non-enzymatic antioxidants from guava and fig fruit filtrate on the motility and viability of Boer buck sperm. Semen was collected using an artificial vagina every four days for ten replications. The treatment extenders were Tris+2.5% egg yolk (P0) and P0+1.487 mg vitamin A and P0+3.958 mg vitamin C per ml/100 ml (v/v) hereinafter referred as P1 and P2, respectively. The experimental design was a completely randomized 3 times 10 a factorial pattern. The variables assessed were individual motility, progessive motility and viability of sperm evaluated every day for 4 consecutive days. Result indicated that, 4 days after extended and stored at 5°C, the average individual motility, progessive motility and sperm viability were 45.2±7.94; 33.0±10.2 and 49.6±7.69 for P1, and 45.4±6.14; 30.4±8.38 and 50.5±5.88 for P2. The data on the three indicators of sperm quality between P1 and P2 did not show a significant difference (P0.05). While for P0 30.6±5.85; 24.6±7.30 and 40.7±5.77, respectively, and were significantly lower (P 0.05) than P0 or P1. It was concluded that semen extended using tris-egg yolk extender supplemented with vitamin A or C could still be used for Artificial Insemination until day 4 after extended and storage at 5°C. It is suggested that research needs to be done to produce a ready to use, simple preaparation, cheap and long lasting instant extender.

In vivo Fertility Assessment of Frozen Thawed Pantja Buck Semen

Background: Pantja is a medium sized, dual purpose Tarai goat breed of Uttarakhand and adjacent area of Uttar Pradesh. Due to the lack of pure breeding buck and uncontrolled cross breeding, population of this breed is reducing continuously. Therefore, for preservation and dissemination of this pure breed, cryopreservation of semen and artificial insemination by using frozen thawed semen can play a crucial role. Methods: Semen samples were collected from four sexually matured Pantja bucks and after preliminary evaluation extended in tris egg yolk based extender. Post thaw semen was examined for microscopical and certain biochemical evaluation. A total 12 does were inseminated and pregnancy diagnosis was conducted at 45 days onwards by ultrasonography. Result: Post thaw seminal attributes were significantly (P£0.01) reduced in comparision to freshly diluted semen. Total six does (50.00%) were pregnant and five (83.33%) of them gave birth to healthy kids. Among these five, three does (60.00%) gave birth to single and two does (40.00%) gave birth to twins.

Effect of clarified tris egg yolk citrate diluter and seminal plasma removal on plasma membrane integrity in cryopreserved Surti buck semen

Effect of clarified tris egg yolk citrate diluter and seminal plasma removal on plasma membrane integrity in cryopreserved Surti buck semen

The effectiveness of Adding Different Sucrose in Tris-Egg Yolk Diluent on the Sperm Fertility of Sexing Swamp Buffalo (Bubalus bubalis)

Spermatozoa sexing requires a diluent that can protect and provide an optimal environment to maintain the quality of spermatozoa. This study targets to determine the effectiveness of adding sucrose to tris-egg yolk diluent on the sperm quality of the swamp buffaloes produced by sexing with the egg albumin method. Semen was collected once a week for 12 consecutive weeks with an artificial vagina from three male buffaloes at the Indonesian Center for Animal Research and Development (ICARD) in April - June 2021. Semen was separated by sedimentation using 10% and 30% egg albumins. Semen from sexing was divided into four diluent treatments, namely 20% tris-egg yolk (TEY), TEY+0% sucrose, TEY+0.2% sucrose, TEY+0.3% sucrose, and TEY+0.4% sucrose. Parameters measured included: motility, viability, intact plasm membrane, and intact acrosome cap of buffalo sperm. The results showed the motility of X sperm: 43.75-47.5%, Y sperm: 40-45%, the viability of X sperm: 78.25-79.75% and Y sperm: 77.25-79.75%, intact plasm membrane of X sperm: 70.5-71.5% and Y sperm: 70.5-71%, and viable acrosome-intact for X sperm: 79-80% and Y sperm: 78.75-79.5%. The addition of sucrose up to 0.4% into tris-egg yolk diluent was proven can maintain motility, viability, and integrity of cell membrane and acrosome of spermatozoa of water buffalo that was sexed with egg albumin.

The influence of L-proline and fulvic acid on oxidative stress and semen quality of buffalo bull semen following cryopreservation.

This study investigates the effects of cryopreservation and supplementation of Azeri water buffalo's semen with proline (Lp) and fulvic acid (FA). Therefore, this study aimed to assess motility parameters, sperm viability, oxidative stress parameters, and DNA damage to detect the optimum concentrations of Lp and FA for buffalo semen cryopreservation. Thirty semen samples of three buffalo bulls were diluted in Tris-egg yolk extender and divided into 12 equal groups including control (C), Lp-10, Lp-20, Lp-40, Lp-60, Lp-80 (containing 10, 20, 40, 60, 80mM L-proline, respectively), FA-0.2, FA-0.5, FA-0.8, FA-1.1, FA-1.4 and FA-1.7 (containing 0.2%, 0.5%, 0.8%, 1.1%, 1.4% and 1.7% fulvic acid, respectively). The velocity parameters, TM and PM were improved by FA-1.7, FA-1.4, Lp-40 and Lp-60 groups compared to the C group but no significant difference was found regarding the amplitude of lateral head displacement and straightness compared to the control groups. The percentage of sperm viability and PMF were increased by FA-1.7, FA-1.4, FA-1.1, Lp-40 and Lp-60 groups compared to C group, while in terms of sperm DNA damage FA-1.7, FA-1.4, FA-1.1, Lp-10, Lp-20, Lp-40 and Lp-60 groups showed better results compared to C group. The results also showed that FA-1.7, FA-1.4, FA-1.1, Lp-20, Lp-40 and Lp-60 groups could improve TAC, SOD, GSH and decrease MDA levels. Also, FA-1.7, FA-1.4, Lp-20 and Lp-40 groups could improve GPx levels but just FA-1.7, and Lp-40 groups could improve CAT levels compared to C group. Thus, it can be concluded that L-proline and fulvic acid supplementations can improve the quality parameters of post-thawed buffalo bull semen.

Aqueous Extract ofMoringa OleiferaLam. Leaves Added to Freezing Extenders Damages Goat Sperm Membranes

BACKGROUND:Semen cryopreservation is a biotechnology used frequently in animal production; however, there are some obstacles, such as those caused by high levels of reactive oxygen species (ROS).Moringa oleifera(MO) is known as a potent source of antioxidants and might be an important adjuvant .OBJECTIVE:The objective of this study was to determine the effect of different concentrations of MO extract supplementation on goat semen cryopreservation efficiency.MATERIALS AND METHODS:Ejaculates (n=6) from four goat breeders were pooled and diluted in skimmed milk (SM) or Tris-egg yolk (TEY)-based extenders and supplemented with different concentrations of MO extract (0, 1, 2 and 5 mg/mL). After the freeze-thaw cycle, sperm kinetics and viability were assessed.RESULTS:With the SM extender, straightness, wobble and plasma membrane integrity were lower than in the control group (ρ &lt; 0.05). With the TEY extender, wobble was lower in with 5 mg/mL MO extract than in the control group (ρ &lt; 0.05). As regards sperm ultrastructure, evaluated by SEM, the MO extract, regardless of the diluent used, damaged the membrane of sperm cells in a dose-dependent manner.CONCLUSION:The addition of aqueous extract of MO leaves in both diluents at all concentrations tested affects the parameters of sperm progressivity and damages the plasma membrane in a dose-dependent manner.

Buffalo anti-PDC-109 antibodies improve the semen quality profiles and in-vitro zona binding index and minimize the cryoinjury of sperm in cryopreserved buffalo semen

Objective: To optimize the concentration of PDC-109 protein in semen for higher cryopreservability and fertility by sequestration or neutralization of PDC-109 by its antibodies (anti-PDC-109 antibodies) in bubaline species. Methods: PDC-109 protein was purified by applying two-step chromatography procedures. Purified protein was injected in rabbits to raise antibodies. These raised anti-PDC-109 antibodies were used in neutralization or sequestration of PDC-109 in in-vitro model. Ejaculates were collected from buffaloes and splited for four groups. Group 1 received egg yolk Tris glycerol extender, without anti-PDC-109 antibodies, while group 2 to 4 received anti-PDC-109 antibodies 266 μg/mL, 80 μg/mL, and 26 μg/mL in Tris-fructosecitrate buffer, respectively. Semen quality parameters viz., forward progressive motility, viability, total morphological abnormality, acrosomal integrity, plasma membrane integrity, cryoinjury and in-vitro zona binding index were evaluated. Results: Semen quality parameters of neat semen were within the normal range of bubaline species. Sperm motility, livability, acrosomal integrity, plasma membrane integrity, and cholesterol content of sperm were decreased and total sperm abnormality was increased significantly in post-thaw semen compared to those in pre-freeze and fresh semen (P&lt;0.05). Semen in group 2 had higher sperm motility, livability, acrosomal integrity, plasma membrane integrity, and cholesterol content of sperm and lower total sperm abnormality significantly compared to those in group 1, 3 and 4 at pre-freeze and post-thaw stages (P&lt;0.05). Conclusions: Sequestration or neutralization of PDC-109 by its antibodies significantly improves pre-freeze, and post-thaw semen quality parameters and in-vitro zona binding index with simultaneously reducing cryoinjury or cryodamage in the sperm of bubaline species.

Effect of Addition of Green Tea Extract (Camellia sinensis) in Egg Yolk Tris Diluter on Spermatozoa Quality in Bali Cattle (Bos sondaicus) After Freezing

This study aimed to determine the effect of adding 96% ethanol extract of green tea (Camellia sinensis) in egg yolk tris diluter on the quality of spermatozoa of Bali cattle (Bos sondaicus) after freezing. The samples were divided into four different treatment groups, which contained semen sample + 13% glycerol + each egg yolk tris diluter + green tea extract (P0) 0 mg/100 ml, (P1) 0,05 mg/100 ml, ( P2) 0,10 mg/100 ml, and (P3) 0,15 mg/100 ml. The variables observed included motility, viability, and spermatozoa abnormalities. Data were analyzed using OneWay ANOVA and continued with Duncan's test (p&lt;0,05). The results showed that the P3 group had motility of 49,00 ± 2,15, viability of 61,92 ± 1,70, and abnormality of 4,24 ± 2,16 significantly compared to the other groups. In conclusion, the P3 group with the addition of 0,15 mg/100 ml of green tea could increase the motility, viability, and abnormal spermatozoa of Bali cattle.

Effect of Vitamin D3 on some Antioxidant Parameters in Chilled Semen in Awassi Ram.

To sustain the viability of the sperm of farm animals, the sperm is chilled. However, reactive oxygen species (ROS) may damage it, resulting in oxidative stress and decreased sperm viability. This study aimed to assess the various concentrations of vitamin D3 as antioxidants in the chilled sperm of Awassi. This study was performed on 23 ejaculates from three Awassi rams. The samples were combined, diluted with Tris-egg yolk extender (1:10), and then divided into aliquots. Aliquots were treated with three vitamin D3 concentrations (T1=0.02, T2=0.004, and T3=0.002 g/ml) and one control without the addition of vitamin D3. The experimental and control groups were chilled to reach 5 ºC. Following treatment, the samples were centrifuged at 2,000 RPM for 20 min at 0 and 72 h after the treatment. Until evaluation, the seminal plasm was stored in a freezer at 20 ºC. In this study, the antioxidant activity of vitamin D3 was evaluated using malondialdehyde (MDA), ROS, total antioxidant capacity (TAC), superoxide dismutase (SOD), and catalase (CAT). The SAS software was used to analyze variance on repeated measures with a single factor. The results indicated that the TAC and SOD were considerably higher in T1, compared to that in T0, T1, and T2. In addition, CAT was considerably higher in T2 than in T0, T1, and T3. However, ROS and MDA did not differ significantly among the experimental groups. Despite the absence of a statistically significant difference among experimental groups, MDA decreased quantitatively on T1, relative to other experimental groups. In conclusion, a deficiency in vitamin D3 has a potential antioxidant capability, introducing a novel method for extending sperm storage.

Studi Literatur Efektivitas Penggunaan Berbagai Jenis Pengencer yang Ditambahkan Antioksidan Terhadap Kualitas Semen Kambing (Capra aegagrus hircus)

Goats are ruminants that have considerable potential. The livestock population data in the province of East Nusa Tenggara (NTT) has continued to increase from 2004 to 2018. The Artificial Insemination (AI) program has a function to improve genetic quality and livestock production. Extender is a mixture of various materials that can maintain the quality of spermatozoa in animal semen. Antioxidants to prevent the emergence of oxidant compounds in excess. The purpose of this literature study was to determine which semen extender has effectiveness in improving the quality of goat semen and the active substance that acts as an antioxidant as one of the substances that can improve the quality of the extender. The process of this literature study is carried out by searching and collecting various kinds of existing literature using Google Scholar, Google Patent, Pubmed then all existing references are compiled with the Mendeley application. Reference sources obtained are scientific journals, scientific articles/publications, e-books and national and international seminar proceedings by searching for "keywords" the effectiveness of using various types of extenders added with antioxidants on the quality of goat semen collected with a total of approximately 85 references. The results obtained from the literature study carried out are on the effectiveness of the semen extender, namely the more effective commercial extender, namely Andromed®, then the natural extender, namely guava leaves, while the compound extender is TGC+CLC (tris citrate glucose + Cholesterol Loaded). Cyclodextrin). Antioxidant active substances, both commercial and natural, that can increase the quality of goat semen, namely commercial antioxidants are Glutathione + 6 mM, while natural antioxidants are TKT (Egg Yolk Tris) + 4% Carrot Extract. The temperature and storage time of goat semen, the optimal temperature used for the equilibration time temperature is 5℃, at the cryopreservation temperature at -196℃, then at the optimal storage

The Effect of Lycopene Addition on the Semen Quality of Saanen Goats Stored at 5°C for 24 hours

In order to increase the livestock population, a good reproductive process was needed. Lycopene which belongs to carotenoid group has a dark yellow pigment. This study aimed to determine the effect of adding lycopene on the semen quality of Saanen goats. The research design used 4 treatments, P0 (egg yolk tris diluent) as control), P1 (egg yolk tris diluent + 2% lycopene), P2 (egg yolk tris diluent + 4% lycopene), and P3 (yolk tris diluent). egg + 6% lycopene). The quality of semen was observed after 24 hours of storage at 5℃. Macroscopic data were analyzed descreptively and the microscopic one was analyzed using a completely randomized design (CRD). The results showed that the addition of lycopene in egg yolk tris diluent had a significant (P&lt;0.05) effect on the semen quality of Saanen goats. The results showed that the motility of spermatozoa in each treatment group (P0, P1, P2, and P3) was 60.1 ± 5.7%, 65.2 ± 4.3%, 62.4 ± 5.2%, and 60.6 ± 4.7%, respectively, viability were 65.3 ± 4.8, 73.5 ± 4.2, 70.4 ± 3.8, and 66.1 ± 5.1, respectively, abnormality were 17 ± 1.8%, 16 ± 1.8%, 18 ± 1.9%, and 16 ± 1.7%, respectively, and membrane integrity were 63 ± 3.3%, 70 ± 3.6 %, 67 ± 3.2%, and 62 ± 2.9%, respectively. Based on the results, it could be concluded that the lycopene addition improved the semen quality of Saanen goats compared to controls.

Antioxidant additive melatonin in tris-based egg yolk extender improves post-thaw sperm attributes in Hariana bull

In the study, melatonin, a known antioxidant pineal peptide was used as an additive in the tris-egg yolk glycerol-based semen extender in Hariana bull semen and post-thaw sperm characters were evaluated. In the study, Group I was a control without melatonin; and Group II, III, and IV were having 0.5 mM, 1 mM, and 2 mM melatonin/80 × 106 spermatozoa, respectively were treatment groups. Thirty-two semen ejaculates from 4 Hariana bulls were processed for freezing and post-thaw sperm characteristics were evaluated. Sperm motility, velocity, the viability with intact membrane, and total antioxidant capacity were markedly (P < 0.05) improved in Group IV compared to all other groups. The lipid peroxidation and total protein carbonylation were substantially (P < 0.05) decreased in Group IV compared to all other groups. The population of cryocapacitated, acrosome-reacted, and apoptotic-like spermatozoa were significantly (P < 0.05) decreased in Group IV. Further, the relative band intensity of 74 kDa protein and percent of spermatozoa showing positive immune reactivity to tyrosine-phosphorylated proteins was decreased in Group IV. The progesterone-receptor ligand binding, in vitro capacitation response, and Vanguard distance were markedly (P < 0.05) improved in Group IV. In summary- Group IV having 2 mM melatonin was found to be optimal in providing cryoprotective effects to Hariana bull spermatozoa after freezing-thawing and can be suitably used as a semen additive during semen cryopreservation.

COMPARISON BETWEEN RAPID AND SLOW CRYOPRESERVATION PROTOCOLS FOR RAM SEMEN

Information regarding adequate freezing protocols of ram semen considering freezing distance and time during cryopreservation has not been adequately reported. Therefore, this study aimed to compare two freezing protocols for Najdi ram's semen. In the rapid freezing protocol, the straws were frozen at 5 cm over the surface of liquid nitrogen for 15 minutes. While in the slow freezing protocol, the straws were frozen at 8 cm over the surface of liquid nitrogen for 20 minutes. The semen was collected from five rams and extended with tris egg yolk glycerol cryodiluent. The extended semen was chilled slowly to 50C within two hours and equilibrated for two hours before being frozen on the liquid nitrogen vapor and cryopreserved at -1960C. There was no significant (P &gt;0.005) effect of the freezing protocol on the sperm’s total motility, plasma membrane integrity, DNA integrity, and abnormalities. However, the vitality, fast progressive motility, straight-line velocity, average pathway velocity, linearity, and wobble were significantly higher in rapid freezing than in slow freezing protocol. In conclusion, cryopreservation of ram semen using rapid freezing (5 cm for 15 mins) protocol was better than slow freezing (8 cm for 20 mins) protocol regarding post-thawing semen quality. Key words: freezing protocol; cryopreservation; egg yolk; ram; semen

Comparative Studies on the Effect of Tris -Egg Yolk and Soya Milk Based Semen Extender on various Semen Parameters for Effective Long Term Cryopreservation

Aim: To analyze the most effective Novel semen extender for long time cryopreservation among Tris egg yolk extender and Soya milk extender. Materials and Methods: The sample size was calculated using clinCalc.com with an alpha error-threshold at 0.05, 95 % confidence interval, G power at 80 %, and enrolment ratio 1. The study was repeated 14 times (N=14) and was measured using statistical Independent Samples Test. The study contained a total sample size of 28 with 2 different groups. Tris egg yolk and soya milk based novel semen extenders were added to the collected semen samples and few drops of the sample was placed on a makler counting chamber which was used for analyzing the concentration, sperm motility and sperm count of the semen samples. Results: The Tris egg yolk based semen extenders were better than the soya milk based semen extenders. Estimated sperm motility and sperm count were found to be P&gt;0.05, revealing statistical insignificance. Conclusion: The sperm parameters were preserved well in Tris egg yolk based semen extender when compared with soya milk based semen extender with respect to sperm count and sperm motility.

Determination of Alternative Diluents for Preservation of Duroc Boar Semen Through Modification of Various Semen Diluents

The quality of porcine semen deteriorates very rapidly during the semen preservation process. The aim of this study is to find alternative diluents for Duroc semen preservation by changing different semen diluents. Two Duroc pigs, ± 3 years old, were trained twice weekly for semen collection. Immediately after collection, sperm were examined macroscopically and microscopically. Semen with sperm motility ≥ 70%, sperm concentration ≥200 x 106 cells/ml, and abnormality ≤20%) was diluted with six diluents, namely: Beltsville Thawing Solution® (BTS), Durasperm, Citrate Egg Yolk (CEY), Tris Egg Yolk (TEY), CEY + Olive Oil (CEYO) and TEY + Olive Oil (TEYO) in the ratio of one part semen and four parts diluent. The liquid sperm was stored at 18-20 oC and examined every eight hours for 80 hours. The result shows that sperm motility, viability and plasma membrane integrity of Duroc semen with CEYO and TEYO are comparable to BTS. No significant difference was found in sperm abnormalities. The results suggest that citrate egg yolk and Tris egg yolk in combination with olive oil are suitable as alternative diluents for the preservation of porcine sperm. Further studies are needed to determine the most suitable olive oil concentration for the preservation of porcine sperm of different breeds.