scholarly journals Pengaruh Perbedaan Konsentrasi Gliserol dalam Susu Skim Kuning Telur untuk Proses Penyimpanan Sperma Beku terhadap Motilitas dan Viabilitas Spermatozoa Ikan Patin (Pangasius pangasius) [Effect Of Different Glycerol Concentration In Skim Milk And Egg Yolk Diluter For Storage Process Of Frozen Semen On Sperm Motility And Viability Of Catfish (Pangasius pangasius)]

2019 ◽  
Vol 6 (1) ◽  
pp. 1
Author(s):  
Bagus Rizki Novianto, Sudarno, Endang Dewi Masithah

Abstract Needs of catfish seed always increase every years, especially for farming activities. Fulfillment demand of seed in large quntities and continuing are the main obstacle in the production process of catfish. This can be overcome by the process of cryopreservation. This process requires diluents and cryoprotectants to maintain the fertility of spermatozoa. Skim milk and egg yolks diluents has been used on fish because it was applied to carp and showing a good result of sperm motility and fertilization rate. Glycerol as cryoprotectant was reported effective to be used for European Catfish than the other kind cryoprotectant. This study aimed to determine the effect of different concentrations of glycerol in the diluent skim milk and egg yolk on the motility and viability of catfish (Pangasius pangasius) spermatozoa after freezing. This research method is the experiment with completely randomized design (CRD) as the experimental design. The treatment used is different glycerol concentrations, P1 (11%), P2 (13%), P3 (15%), and P4 (17%) and each of the treatment was repeated 5 times. The main parameters measured were motility (%) and viability (%) of spermatozoa. Supporting parameters include odor, color, pH, viscosity, sperm concentration and catfish (Pangasius pangasius). Analysis of the data using Analysis Of Variance (ANOVA) and to determine the best treatment performed Duncan's Multiple Range Test with 95% confidance interval. The results showed that the effect of the concentration of glycerol in the diluent skim milk and egg yolk was not significantly (P> 0.05) on the motility and viability of catfish (Pangasius pangasius) spermatozoa after freezing. Further testing needs to be done on the level of fertility of frozen sperm, as well as the growth rate of seeds produced from sperm frozen catfish (Pangasius pangasius) products

2013 ◽  
Vol 25 (1) ◽  
pp. 185
Author(s):  
M. M. Seshoka ◽  
M. L. Mphaphathi ◽  
F. V. Ramukhithi ◽  
T. R. Netshirovha ◽  
C. Hlungwani ◽  
...  

There are bull shortages in South African poor rural areas. Artificial-insemination technology could play a significant role on breeding emerging farmer’s cattle. The objective of this study was to compare glycerol concentrations (0, 4, 8, or 12%) during freezing of Nguni bull semen to conduct AI in different villages. Semen was collected by electro ejaculator from 2 Nguni bulls of known and proven fertility. Collected semen samples were kept in a thermo flask (37°C) and transported to the laboratories within 10 min after collections. Semen samples were pooled and evaluated by Sperm Class Analyser® and allocated randomly per treatment. Semen was then diluted (1 : 2 v:v) with egg-yolk citrate extender supplemented with either 0% (negative control), 4, 8, and 12% of glycerol concentration or AndroMed® (positive control). Semen samples were equilibrated for 4 h at 5°C. After equilibration period, samples were loaded into 0.5-mL straws and placed horizontally into the controlled rate (–5, –8, –10, –12, –15, –25, –35°C min–1) from 5°C until target temperature of –80°C is reached. The frozen semen straws were stored in a liquid nitrogen tank (–196°C) until thawing. Treatment means were separated using Fisher’s protected t-test least, and data are presented as mean ± SD. There was a significant differences (P < 0.05) between raw total sperm motility (83.3 ± 19.3) and frozen–thawed sperm with either 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), or 12% (61.5 ± 4.7) of glycerol and on AndroMed® (27.7 ± 17.8) group. Regardless of the glycerol concentrations used, the freezing-thawing process reduced (P < 0.05) the Nguni total sperm motility rate compared to uncryopreserved sperm (83.3 ± 19.3). In conclusion, egg-yolk citrate extender supplemented with 12% glycerol yielded a better (P < 0.05) total sperm motility rate (61.5 ± 4.7) as compared with the 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), and AndroMed® (27.7 ± 17.8) group. Further studies are required to test other levels of glycerol concentrations (>12%) on freezing Nguni semen and conducting AI.


2018 ◽  
Vol 15 (2) ◽  
pp. 215-218
Author(s):  
S Deori

A study was carried out to study the effect of freezing on cryosurvival of yak semen. Artificial insemination in yak is still in infancy. Semen cryopreservation and use of artificial insemination can be applied in yak husbandry for conservation and rapid multiplication of superior germplasm. Semen was collected from four adult yak bulls using artificial vagina method managed under uniform conditions. A total of 40 ejaculates comprising of 10 ejaculates each bull were collected following twice a week schedule and evaluated for fresh semen characteristics. The fresh yak semen characteristics viz. ejaculate volume (ml), mass activity (0-4), initial sperm motility (%), sperm concentration (x 106/ml), live sperm (%), sperm abnormality (%) and intact acrosome (%) were 3.10 ± 0.18, 3.53 ± 0.96, 83.89 ± 2.87, 1180.22 ± 42.32, 77.63 ± 4.23, 8.45 ± 3.33 and 93.61 ± 3.78 respectively. The ejaculates were diluted (1:10) with Tris extender consisting of 6.4 ml glycerol and 20 ml of fresh egg yolk. Straws were equilibrated at 5°C for 4 hours followed by exposure to liquid nitrogen vapour for 10 minutes and finally transferred to liquid nitrogen container for storage. The cryosurvival rate was studied after 7 days of storage in liquid nitrogen. The frozen semen was thawed in warm water (37°C) for 30 seconds for evaluation. Mean values of postthaw sperm motility (%), live sperm (%) and intact acrosome (%) in yaks were 55.67 ± 4.67, 65.62 ± 3.23 and 89.26 ± 3.67 respectively. In conclusion, yak semen has a better cryosurvival while freezing in tris extender with 6.4 per cent glycerol and 20 per cent egg yolk following an equilibration period of 4h.SAARC J. Agri., 15(2): 215-218 (2017)


2004 ◽  
Vol 16 (2) ◽  
pp. 127 ◽  
Author(s):  
G. Brogliatti ◽  
G. Barreiro ◽  
G. Larraburu ◽  
A. Laborde

Flow citometry cell sorting has been proven successfully to separate X and Y sperm; however, the technology is still too stressfull for the viability of the sorted semen. The objective of this study was to evaluate nonsexed and sexed frozen sperm motility characteristics using a CASA technology. Ejaculates from 4 different bulls (3 Holstein and 1 Angus) were collected, and processed as split non-sexed and sexed semen samples using Tris egg yolk extenders. X and Y sperm were separated using a high-speed sorter (SX Moflo). Cryopreservation was done at the same time under appropiate conditions using a programmed cryochamber. Thawing procedure was done at 37°C for 30s and a sample of each straw was placed in the evaluation chamber. The experiment was repeated twice and two chambers with 30 observations each were analyzed each time. Mean and standard deviation of each characteristic were calculated, compared and analyzed statistically. The sperm concentration was determined by means of a burker counting chamber. Sperm quality was determined at 0h after thawing, and later at 1h, 2h and 3h after incubation in a glass tube at 30°C. The following sperm motility parameters were determined with the Hamilton Thorne (HTM-ceros 12.1) on at least 1000 spermatozoa: velocity average path (VAP), velocity straight line (VSL), amplitude lateral head (ALH), beat cross frequency (BCF), straightness (STR), linearity (LIN), and percentage of progressively motile spermatozoa (PMS). Linearity of nonsexed spermatozoa was 53±3.5, 47±0.8, 43±7.8 and 42±4.5 for the 0h and the 3 test incubation times and 49.5±3.7, 51.2±3.7, 43.3±7.8 and 44.5±7.6, respectively, for sexed semen. There were no significant differences (P&gt;0.05) in the progressive velocity, track speed and linearity between sexed and nonsexed semen. The percentage of static cells was 33%, 30%, 47% and 50% for the 0h and the 3 test incubation periods; however, the percentage of static cells for the sexed semen was 53%, 71%, 77% and 82%, respectively. Results from the analysis indicate a significant increase (P&lt;0.01) in the number and the percentage of static cells with time. The lateral amplitude of sperm motility for nonsexed semen was 5.9±0.5, 6.8±0.8, 6.0±0.4 and 5.1±0.7, and for sexed semen 6.6±0.7, 6.8±0.4, 6.4±0.4 and 5.5±1.7, respectively. The percentage of progressively motile sperm was significantly different at 0 time 49.7±4.9 and 23.1±4.9 for nonsexed and sexed semen respectively. After 3 hours of incubation the percentage of progressively motile sperm was 38.7±10.2 and 3.7±3.2 for nonsexed and sexed semen, respectively. In conclusion, sexed frozen semen seems to have characteristics similar to those of normal nonsexed semen. However, a significant decrease in the percentage of progressively motile cells could affect pregnancy rates. More research needs to be done to detect differences between bulls and cryoprotectans.Research supported by Centro Genetico Bovino de EOLIA sa Argentina.


2020 ◽  
Vol 13 (11) ◽  
pp. 2422-2428
Author(s):  
Ani Atul Arif ◽  
Tulus Maulana ◽  
Ekayanti Mulyawati Kaiin ◽  
Bambang Purwantara ◽  
Raden Iis Arifiantini ◽  
...  

Background and Aim: Indonesia has two National Artificial Insemination centers and 17 Regional Artificial Insemination Centers. The frozen semen production techniques differed between the centers, including the type of diluent and semen dilution technique. The aim of the research was to compare the quality of frozen Limousin bull semen diluted using different techniques. Materials and Methods: Semen was collected from three sexually mature Limousin bulls using an artificial vagina. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen that had >70% motile sperm and <20% sperm abnormality was divided into three tubes and diluted with skim milk-egg yolk (SMEY) using three different dilution techniques: One-step dilution (100% SMEY with 8% glycerol) at room temperature ([RT] 20°C until 25°C) two-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol after 1 h stored at 5°C); and three-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol added twice at 1 h and 1.5 h after being stored at 5°C). The diluted semen was loaded into 0.25 mL mini straws, equilibrated, and frozen using a freezing machine. Sperm motility, viability, membranes, DNA integrity, and concentrations of malondialdehyde (MDA) and aspartate aminotransferase (AST) enzymes were evaluated after thawing. Results: The results showed that there were no significant differences in sperm motility and DNA integrity between dilutions (p>0.05). However, sperm viability and membrane intactness of one-step dilutions were higher than those of three-step dilutions. The concentrations of MDA and AST enzymes of sperm in one-step dilutions were lower than those of three-step dilutions (p<0.05). Conclusion: It was concluded that the one-step-dilution technique was better than three-step dilution for cryopreservation of Limousin bull semen.


2021 ◽  
Vol 3 (3) ◽  
pp. p12
Author(s):  
Cut Intan Novita ◽  
Faqihuddin Nasution ◽  
Eka Meutia Sari

The process of semen freezing causes an increase in free radicals concentration which can damage spermatozoa. The addition of natural ingredients in semen diluent is expected to solve this challenges. One of the natural ingredients that can be used is jamblang (Syzygium cumini) leaves. The objective of the current study was to investigate the quality of spermatozoa in Aceh cattle which was added with jamblang leaves extract in skim milk- egg yolk extender during pre-freezing and post-thawing. This study applied Completely Randomized Design (CRD) with 5 treatments and 5 replications. The treatments consisted of J0 = skim milk-egg yolk; J1 = skim milk-egg yolk + jamblang leaves extract 0.2%; J2 = skim milk-egg yolk + jamblang leaves extract 0.4%; J3 = skim milk-egg yolk + jamblang leaves extract 0.6%; and J4 = skim milk-egg yolk + jamblang leaves extract 0.8%. The parameters observed in this study were the percentage of motility and viability of frozen semen of Aceh cattle. The data obtained were analyzed using Analysis of Variance (ANOVA) and if differences were found, then it would be continued with Duncan's Multiple Distance test. The results showed that the addition of jamblang leaves extract in egg yolk skim milk significantly affected the percentage of motility during pre-freezing and post-thawing, significantly affected spermatozoa viability during pre-freezing and significantly affected the spermatozoa viability during post-thawing. J3 treatment (jamblang leaves extract 0.6 gram/100 ml) it should be higher than the other treatment, where the percentage of motility at pre-freezing and post-thawing were 55.48% and 52.71%, respectively, and the percentage of viability during pre-freezing and post-thawing were 56.59% and 53.94%, respectively. It was concluded that the addition of jamblang leaves extract in the skim milk-egg yolk extender affected the percentage of spermatozoa motility and viability of Aceh cattle during pre-freezing and post-thawing.


2016 ◽  
Vol 1 (2) ◽  
pp. 6-13
Author(s):  
J. Junaedi ◽  
Raden Iis Arifiantini ◽  
Cece Sumantri ◽  
Asep Gunawan

On the freezing semen chicken cryoprotectants required to overcome the damage of spermatozoa due to cold shock. This study aims to get the best concentrations of cryoprotectants glycerol concentration of 5%, 7% and 9% in freezing Sentul chicken semen. The semen used in this study came from three chickens Sentul and be repeated nine times. Semen was collected by  messase methods for three times a week. Semen was evaluated macroscopic and microscopic. Furthermore spermatozoa diluted with egg yolk and the addition of three concentrations of cryoprotectants glycerol (5%, 7% and 9%). Semen diluted 0:25 ml is packed into straw. Then equilibrated at a temperature of 5°C for two hours. After equilibration to evaluate the motility and viability of spermatozoa. Furthermore, frozen in liquid nitrogen vapor for 10 minutes. Frozen semen is then stored in liquid nitrogen containers with temperature -196°C. After 24 hours, semen is thawed at 37°C for 30 seconds. The results showed that the percentage of sperm motility and viability of frozen semen cock Sentul using glycerol cryoprotectants 5% better P (<0.05) compared with the use of glycerol 7% and 9%. The use of glycerol 5% at this stage of equilibration and storage can reduce the damage of spermatozoa in the semen of chicken Sentul. Neither glycerol 5% could increase recovery rate after thawing


2021 ◽  
Vol 19 (2) ◽  
pp. 237-244
Author(s):  
Nguyen Huu Duc ◽  
Pham Thu Giang ◽  
Tran Thi Binh Nguyen ◽  
Nguyen Thi Mai ◽  
Bui Dai Phong

The objective of this study was to determine the semen cryopreservation capacity of BBB bulls in Hanoi-Vietnam. Research conducted on the fresh semen collected from 05 BBB bulls. Results showed that semen color was normal (milky white, ivory white, ivory yellow), semen volume ranged from 6.35 mL to 7.48 mL (P <0.05), initial motility of semen ranged from 80.53% to 82.92% (P <0.05), sperm concentration in semen  ranged from 1.02 x 109 sperms/ml to 1.12 x 109 sperms/mL (P <0.05), abnormal sperm ratio ranged from 6.45% to 8.12% (P <0.05), alive sperm ratio ranged from 76.34% to 82.97% (P <0.05), sperm motility after thawing from straw semen ranged from 71.33% to 75.92% (P<0.05). In conclusion, successfully semen collection from 05 breeding BBB bulls at Hanoi Cattle Breeding Joint Stock Company, semen samples had normal color and good quantity and quality, suitable for production of frozen semen; and semen cryopreservation of straws of the 05 bull BBB semen mentioned at -196oC, sperm motility after freezing-thawing reached the economic and technical norms of 675/2014 of the Ministry of Agriculture and Rural Development.


2015 ◽  
Vol 27 (1) ◽  
pp. 126
Author(s):  
H. S. Martins ◽  
M. F. Brito ◽  
I. B. M. Sampaio ◽  
R. Stahlberg ◽  
M. R. Souza ◽  
...  

During cryopreservation, the sperm were submitted to an increased generation of reactive oxygen species. Furthermore, because of the large portion of seminal plasma removal, there is a decrease of sperm antioxidant protection. Addition of antioxidants proteins found in seminal plasma, such as lactoferrin (Lf) and catalase (Cat), to the freezing semen extenders could protect the sperm during cryopreservation. Lactoferrin is a transferrin, which prevents the hydroxyl radicals generation, and Cat plays an antioxidant role. The aim of this study was to determine the effects of Lf and Cat supplementation to the INRA 82 freezing extender (Battelier et al. 1997) on sperm motility parameters and membrane functionality of stallion frozen semen. Semen from 6 stallions was collected with an artificial vagina, diluted with Kenney extender (1 : 1), and centrifuged (500 × g, 10 min). The supernatant was discarded, and sperm number per milliliter was calculated. Semen was resuspended with 3 extenders to 100 × 106 sperm mL–1. The treatments were distributed in (F1) control, INRA 82 freezing extender (Battelier et al. 1997), (F2) F1+ 500 μg mL–1 lactoferrin, and F3) F1 + 200 IU mL–1 catalase. Semen samples were packaged in 0.5-mL straws and cooled to 5°C (0.27°C min–1). For semen freezing, the straws were laid over the LN vapor for 20 min and plunged into the LN. The straws were thawed at 37°C for 30 s. Motility parameters of frozen semen were determined using a computer sperm cell analysis, and sperm membrane functionality was assessed by the hyposmotic swelling test (Lagares et al. 1998). The data were analysed using Friedman test using stallion as a block. A probability of P < 0.05 was considered significant. There was no significant difference between the percentage of total sperm motility (median, minimum-maximum value; F1: 29.9, 11.0–82.7; F2: 49.8, 7.7–55.2; F3: 39.8, 5.7–92) and progressive sperm motility (F1: 7.1, 3.2–23.3; F2: 13.4, 2.6-22.4; F3: 15.6, 1.1–29.6), and functional sperm membrane (F1: 26.7, 14.7–56.2; F2: 50.5, 15.7–61.7; F3: 46.6, 13.8–50.9) with regard to the treatment. However, the velocity parameters: velocity average path (F1: 29.3, 22.1–33.80; F2: 34.6, 24.8–44; F3: 35.7, 18.2–42.6), velocity curvilinear (F1: 36.9, 30.5–45.1; F2: 42.5, 34.7–51; F3: 44.6, 25.5–50.9), and velocity straight line (F1: 23.4, 17–3.60; F2: 28.9, 18.8–38.2; F3: 26.6, 13.6–37.2) in the treatment with Lf (F2) were higher compared with the control (F1; P < 0.05). These results corroborate with studies reporting the lack of positive effect on equine sperm motility when antioxidants were added to skim milk-based extenders. Although the addition of Lf or Cat to skim milk-based extenders did not improve the motility sperm characteristics and sperm membrane functionality, more studies about the positive effect of Lf on the velocity parameters are necessary. Lactoferrin could then play an important role on the oxidative metabolism, which provides energy to the sperm movement.Acknowledgments to the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil, for the financial support.


1972 ◽  
Vol 52 (1) ◽  
pp. 65-72 ◽  
Author(s):  
L. M. SANFORD ◽  
G. J. KING ◽  
J. W. MACPHERSON

Boar and bull spermatozoa were diluted in a skim milk–egg yolk–glucose extender containing 0, 7.5, or 15% glycerol (v/v) and incubated aerobically for 6 hr at 37 C. Other partially diluted boar semen samples were cooled to 5 C. Glycerol was added to a final concentration of 0, 7.5, and 15%. Samples were frozen to −79 C, rewarmed, and incubated for 3 hr at 37 C. The presence of glycerol in the extender depressed (P < 0.01) the oxygen uptake by nonfrozen boar and bull spermatozoa during the 6-hr incubation period. The reduction of oxygen uptake by semen samples increased as the level of glycerol in the extender increased. There was a corresponding decrease (P < 0.01) in the number of motile cells at the conclusion of the incubation period. Glycerol appeared to have more of a detrimental effect on boar spermatozoan oxygen uptake. The rate of oxygen uptake by boar semen samples postfreezing was extremely depressed, suggesting that spermatozoa surviving the freezing process metabolize at a much lower rate than normal. Active progressive motility of most of the surviving boar spermatozoa ceased within 1–2 hr of incubation under the in vitro conditions of this experiment.


2020 ◽  
Vol 7 (2) ◽  
pp. 96
Author(s):  
Rudi Irvanto ◽  
Hardijanto Hardijanto ◽  
Widya Paramita ◽  
Suherni Susilowati ◽  
Tita Damayanti L ◽  
...  

Quality of spermatozoa motility and viability from rejected limousin bull semen diluted with skim milk egg yolk sitrat added with various levels of glucose. Glucose level used were 0%, 0,5%, 1,5%, 2,5%, and 3,5%. Writer was using on four years old Limousine bull. Bull semen used in this research was bull rejected semen with bellow 70% motility. Semen observation was done at 0 hour, 24 hours and 48 hours. Research design used in this study was completely randomized design with faktorial pattern with 5 replicates. Highest result in motility this research was showed at 24 hours with 30% value in glucose 2,5% treatment and 48 hours with 10% value. The lowest result showed in glucose 0% treatment at 24 hours and 0% at 48 hours. Highest result in viability showed on glucose 2,5% treatment with 62,6% value at 24 hours and at 48 hours with 53,4% value. Lowest result in viability showed on glucose 0% treatment with 44,2% value at 24 hours and 31,4% value at 48 hours.


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