10CASA EVALUATION OF SEXED AND NON-SEXED FROZEN BULL SEMEN

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>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<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.

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21 Effect of egg yolk extracted low-density lipoprotein on cryopreserved Nguni bull semen

Reproduction Fertility and Development ◽

10.1071/rdv31n1ab21 ◽

2019 ◽

Vol 31 (1) ◽

pp. 136

Author(s):

M. M. Tshabalala ◽

K. A. Nephawe ◽

M. L. Mphaphathi ◽

C. M. Pilane ◽

T. L. Nedambale

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Egg Yolk ◽

Semen Parameters ◽

Dna Integrity ◽

Low Density ◽

Bull Semen ◽

Frozen Semen ◽

Live Sperm

Egg yolk has been reported to have a beneficial effect on sperm quality following cryopreservation, and this led to its widespread use in semen extenders. However, egg yolk contains substances that inhibit respiration of sperm cells and diminish their motility rate. Moreover, it also contains low-density lipoproteins (LDL) that have a protective effect on sperm during the cryopreservation process. The objective of this study was improve cryopreservation of Nguni bull semen using egg yolk low-density protein. A total of 25 ejaculates were collected from 5 Nguni bulls aged 4 to 5 years using an electroejaculator during the natural breeding season. Collected raw semen samples were transported to the laboratory and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity before dilution. Semen was randomly diluted with a sodium citrate extender supplemented with 20% egg yolk (control) and with 6, 8, 10, and 12% LDL concentrations. The diluted semen sample groups were equilibrated for 4h at 5°C. Following equilibration, semen was loaded into 0.25-mL straws and frozen in a controlled-rate programmable freezer. The groups of semen straws were then plunged into LN and transferred into LN tanks (−196°C) for storage. The frozen semen straws per treatment group were thawed at 37°C and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity. Data were analysed with ANOVA using Stata V12 statistical software (StataCorp., College Station, TX), and treatment means were separated using Fisher’s protected t-test at the significant level of P<0.05. Sperm motility and membrane integrity were significantly higher (P<0.05) in frozen-thawed semen diluted with 8% LDL compared with the other concentrations. However, 6 and 8% LDL resulted in a significantly higher (P<0.05) live sperm, DNA, and acrosome integrity. Frozen-thawed semen diluted with 10 and 12% LDL resulted in the lowest percentages of sperm motility, live sperm, plasma membrane, acrosome, and DNA integrity following cryopreservation. In conclusion, extender containing 8% LDL resulted in improved Nguni bull semen parameters such as sperm motility, viability, plasma membrane, acrosome, and DNA integrity following cryopreservation. Further studies are required to determine the fertilizing capacity of semen diluted and frozen with LDL in vitro and in vivo.

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76 THE EFFECT OF GLYCEROL CONCENTRATIONS IN EGG-YOLK CITRATE EXTENDER ON THE QUALITY OF CRYOPRESERVED NGUNI BULL SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab76 ◽

2013 ◽

Vol 25 (1) ◽

pp. 185

Author(s):

M. M. Seshoka ◽

M. L. Mphaphathi ◽

F. V. Ramukhithi ◽

T. R. Netshirovha ◽

C. Hlungwani ◽

...

Keyword(s):

Sperm Motility ◽

Rural Areas ◽

Egg Yolk ◽

Positive Control ◽

Negative Control ◽

Glycerol Concentration ◽

Bull Semen ◽

Frozen Semen ◽

Thawing Process ◽

Motility Rate

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.

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62 EFFECTS OF VARYING GLUTATHIONE CONCENTRATIONS IN SEMEN EXTENDER ON THE QUALITY OF FROZEN–THAWED CANINE SPERM

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab62 ◽

2014 ◽

Vol 26 (1) ◽

pp. 145

Author(s):

K. Ogata ◽

B. Sarentonglaga ◽

M. Yamaguchi ◽

A. Sasaki ◽

Y. Kato ◽

...

Keyword(s):

Sperm Motility ◽

High Speed ◽

Pure Water ◽

Egg Yolk ◽

Motility Index ◽

Term Survival ◽

Genetic Traits ◽

Frozen Semen ◽

Semen Extender

Trans-cervical insemination (TCI) with cryopreserved semen offers a potentially effective approach for breeding canids with specific genetic traits, such as guide dogs for the blind. However, there are technical difficulties in canine sperm cryopreservation, such as the composition of semen extender. The aim of this study was to evaluate the effects of glutathione (GSH) as an antioxidant in the semen extender to improve the quality of frozen-thawed dog sperm. A Tris-egg yolk-citrate extender containing 15.7 mg mL–1 of TRIS, 8.8 mg mL–1 of citric acid, 14.1 mg mL–1 of lactose, 25.4 mg mL–1 of raffinose, 1% (vol/vol) antibiotics, and 20% (vol/vol) egg yolk in ultra-pure water was used as the base medium. Twelve ejaculates were collected from 7 dogs. Each ejaculate was divided into 2 to 5 aliquots and extended with base extender supplemented with 0, 2.5, 5, 7.5, and 10 mM GSH as first dilution. The extended semen was equilibrated for 3 h at 4°C. An equal volume of second extender was added to obtain a final concentration of 6.5% glycerol and sperm per milliliter. The sperm samples were loaded in straws and frozen at 6 cm above the surface of LN2 for 15 min in a styrene foam box and plunged into the LN2. The frozen semen was thawed for evaluation. The motility of sperm was estimated with a phase-contrast microscope and the motile patterns were classified into the following grades: progressively motile at a high speed (+++), progressively motile at a moderate and low speed (++), motile without progression (+), and immotile (–). Then, the sperm motility index (SMI) was determined from the following formula as described previously (Iritani et al., 1975), with some modifications: the percentage of (+++) sperm + the percentage of (++) sperm × 0.75 + the percentage of (+) sperm × 0.5. Sperm motility and the SMI were determined at 0, 1, 2, 3, 4, 12, and 24 h after thawing. Acrosome status was evaluated at 4 h after thawing. Lipid peroxidation (LP) levels at 0 and 12 h after thawing were used to examine the antioxidant ability of GSH. Trans-cervical insemination was carried out on 5 bitches to evaluate the fertility of GSH-treated sperm. The TCI were performed nonsurgically with a laparoscope and deposited 2 mL of semen through a catheter. Each bitch was inseminated 1 to 2 times during oestrus. Data were analysed using ANOVA with the Tukey-Kramer method. We found that the rate of (+++) sperm in the 5 mM GSH group was higher than that in the 0 mM group from 1 to 24 h after thawing (P < 0.05). The SMI was higher in the 5 and 7.5 mM GSH groups than in the 0 mM group (P < 0.05). There were no significant differences in the control and 2.5 and 10 mM GSH groups. Long-term survival was increased in the 5 mM GSH group. Acrosome integrity was higher in the GSH-treated group. The level of LP was lower in the GSH-treated groups at 0 h after thawing (P < 0.05). Trans-cervical insemination with the 5 mM GSH-treated semen resulted in the delivery of 5 pups from 2 bitches. These results indicate that the cryopreservation with 5 mM GSH can improve the motility, viability, and fertility of frozen-thawed canine sperm by its antioxidant effects on the sperm membrane.

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113 Effect of antioxidants on motility and fertility of liquid-stored bovine sperm

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab113 ◽

2020 ◽

Vol 32 (2) ◽

pp. 183

Author(s):

Y. Honkawa ◽

T. Fujikawa ◽

N. Miura ◽

C. Kubota

Keyword(s):

Embryo Development ◽

Sperm Motility ◽

Sperm Concentration ◽

Frozen Storage ◽

Egg Yolk ◽

P Value ◽

Motile Sperm ◽

Progressive Motility ◽

Bovine Sperm ◽

Liquid Storage

It is difficult to maintain sperm in liquid storage for a long time, compared with permanent frozen storage in liquid nitrogen. Antioxidants have been reported to improve the quality and fertility of liquid-stored semen. In this study, we investigated whether antioxidants can extend the motility and fertility of frozen-thawed sperm in liquid storage. Frozen-thawed semen from one Japanese black bull (one ejaculate) was diluted in Tris-citrate-fructose (TCF) diluent with 10% (v/v) egg yolk to a sperm concentration of 1×107 spermmL−1. The antioxidants β-mercaptoethanol (βMe) and glutathione (GSH) were added independently, at various concentrations (0.1, 0.5, 1, and 5mM) to sperm suspensions, and these preparations were compared with Control (no added antioxidant). Sperm suspensions were packaged in centrifuge tubes and placed at 17°C in air and monitored daily until sperm motility had stopped (up to 14 days). Sperm motility was analysed by the Sperm Motility Analysis System (SMAS; Ditect Co. Ltd), and the percentage of progressively motile sperm (straight-line velocity (VSL) of >25μm s−1; Grade A classified by WHO manual), compared with that recorded on Day 0 (100%), was determined each day. For evaluation of fertilizing ability, after incubation in liquid storage for 0, 3, 5, and 7 days, sperm were used for IVF with invitro-matured oocytes (30 oocytes per treatment, three replicates). Embryo development was recorded as the proportion of embryos that reached blastocyst by 8 days after IVF. Data for motility were analysed using one-way ANOVA with Tukey test, and embryo development using chi-squared test. A P-value<0.05 was considered statistically significant. At 7 days, the percentage of progressively motile sperm was significantly higher for 0.5, 1, and 5mM βMe than for Control (30.8%, 48.1%, and 50.3%, vs. 0%, respectively). Treatments with 1 and 5mM βMe maintained some sperm progressive motility for 14 days (9.5% and 14.5%). Treatment with GSH showed the same trend at 7 days (32.2%, 36.3%, and 13.7% for 0.5, 1, and 5mM, vs. 0% for Control); 1 and 5mM GSH maintained sperm progressive motility over 10 days (24.8% and 4.4%). In both antioxidant treatments, embryo development was achieved with sperm stored for up to 5 days (Day 0 vs. Day 5 for 0.1mM βMe: 17.6% vs. 13.8%; for 1.0mM GSH: 26.0% vs. 6.7%; for Control: 17.6% vs. 0%). In this study, antioxidants extended both motility and fertility of frozen-thawed bovine sperm in liquid storage. This result suggests the possibility of application to AI using liquid-stored bovine semen.

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The effect of freezing on cryosurvival of yak sperm

SAARC Journal of Agriculture ◽

10.3329/sja.v15i2.35166 ◽

2018 ◽

Vol 15 (2) ◽

pp. 215-218

Author(s):

S Deori

Keyword(s):

Liquid Nitrogen ◽

Sperm Motility ◽

Artificial Insemination ◽

Sperm Concentration ◽

Egg Yolk ◽

Mean Values ◽

Frozen Semen ◽

Sperm Abnormality ◽

Semen Characteristics ◽

Live Sperm

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)

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Mobile phones affect multiple sperm quality traits: a meta-analysis

F1000Research ◽

10.12688/f1000research.2-40.v1 ◽

2013 ◽

Vol 2 ◽

pp. 40 ◽

Cited By ~ 5

Author(s):

Madhukar Shivajirao Dama ◽

M Narayana Bhat

Keyword(s):

Mobile Phone ◽

Sperm Motility ◽

Sperm Quality ◽

Meta Analysis ◽

Sperm Concentration ◽

Reproductive Age ◽

Quality Traits ◽

Motile Sperm ◽

Mobile Phone Use ◽

Mobile Phone Radiation

As mobile phone usage is growing rapidly, there is a need for a comprehensive analysis of the literature to inform scientific debates about the adverse effects of mobile phone radiation on sperm quality traits. Therefore, we conducted a meta-analysis of the eligible published research studies on human males of reproductive age. Eleven studies were eligible for this analysis. Based on the meta-analysis, mobile phone use was significantly associated with deterioration in semen quality (Hedges’s g = -0.547; 95% CI: -0.713, -0.382; p < 0.001). The traits particularly affected adversely were sperm concentration, sperm morphology, sperm motility, proportion of non-progressive motile sperm (%), proportion of slow progressive motile sperm (%), and sperm viability. Direct exposure of spermatozoa to mobile phone radiation with in vitro study designs also significantly deteriorated the sperm quality (Hedges’s g = -2.233; 95% CI: -2.758, -1.708; p < 0.001), by reducing straight line velocity, fast progressive motility, Hypo-osmotic swelling (HOS) test score, major axis (µm), minor axis (µm), total sperm motility, perimeter (µm), area (µm2), average path velocity, curvilinear velocity, motile spermatozoa, and acrosome reacted spermatozoa (%). The strength of evidence for the different outcomes varied from very low to very high. The analysis shows that mobile phone use is possibly associated with a number of deleterious effects on the spermatozoa.

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258 THE EFFECT OF A PLANT PROTEIN COMPONENT OF MEDIA USED FOR BULL SPERM SEXING ON SPERM MEMBRANE STATUS

Reproduction Fertility and Development ◽

10.1071/rdv20n1ab258 ◽

2008 ◽

Vol 20 (1) ◽

pp. 209

Author(s):

M. Bochenek ◽

Z. Smorag

Keyword(s):

High Speed ◽

Egg Yolk ◽

Spongiform Encephalopathy ◽

Plant Proteins ◽

Total Cost ◽

Bull Semen ◽

Sexed Semen ◽

Sperm Membrane ◽

The Mean ◽

Bull Sperm

The aim of the work was to examine the effect of modified TALP medium (TALP/Pp, Animal Pharma B.V., Hengelo, The Netherlands)—used in the sperm sexing procedure—on bull sperm membrane status. The TALP was modified by replacement of bovine serum albumin (BSA) with a mixture of several plant proteins and soya lecithin (Pp). The Pp component was prepared using a high pressure homogenization process. The TALP/Pp had the same pH and osmotic pressure as the original TALP medium (TALP/BSA). The work was divided into 2 parts: (1) Nine ejaculates collected from 2 bulls (Holstein and Polish Red) were used. Immediately after collection, each ejaculate was split into 2 parts and diluted (1:2) with TALP/BSA or TALP/Pp. The sperm membrane status was examined after 3 days of storage at 15�C. (2) Fifteen ejaculates collected from 5 bulls (Holstein, Polish Red, and Simmental) were used. Each ejaculate was split into 2 parts: the first part was diluted with TALP/BSA, stained, incubated, and sexed according to the XY Inc. bull semen sexing procedure; the second part was diluted, stained, incubated, and collected after sexing into TALP/Pp with no egg yolk addition. In both groups no red food due was used to identify and exclude the dead spermatozoa from the sorted fractions. The sperm sexing procedure was performed with an SX MoFlo high-speed sorter at a speed of 3000–4000 cells/s. After collecting about 10 million spermatozoa, both fractions, X andY, were mixed, centrifuged at 700g for 15 min to concentrate the spermatozoa (20 million mL–1), and the sperm membranes examined. For sperm membrane examination, 'live/dead' samples were stained with SYBR-14/propidium iodide fluorochromes and analyzed by flow cytometry. The data from 20 000 spermatozoa were collected for each sample. The percentage of membrane-intact ('live') spermatozoa was taken for statistical analysis. The mean percentage of live spermatozoa stored for 3 days in TALP/BSA v. TALP/Pp was 25.7% (SD = 7.48) v. 28.58% (SD = 7.04), respectively (P < 0.01). The mean percentage of live spermatozoa in samples of sexed semen was 33.57% (SD = 18.97) for TALP/BSA and 38.51% (SD = 20.22) for TALP/Pp (P < 0.01). It can be concluded that Pp should be considered as a replacement for BSA in the TALP medium used for bull sperm sexing because (1) it results in significantly higher numbers of live spermatozoa after storage and/or sexing; (2) it eliminates a possible source of transmissible diseases (such as bovine spongiform encephalopathy); and (3) it decreases the total cost of the basic media used for the bull sperm sexing procedure.

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Web- and Artificial Intelligence–Based Image Recognition For Sperm Motility Analysis: Verification Study (Preprint)

10.2196/preprints.20031 ◽

2020 ◽

Author(s):

Vincent FS Tsai ◽

Bin Zhuang ◽

Yuan-Hung Pong ◽

Ju-Ton Hsieh ◽

Hong-Chiang Chang

Keyword(s):

Artificial Intelligence ◽

Sperm Motility ◽

Image Recognition ◽

Sperm Quality ◽

Sperm Concentration ◽

Human Sperm ◽

Recognition Algorithm ◽

Motile Sperm ◽

Home Testing ◽

System P

BACKGROUND Human sperm quality fluctuates over time. Therefore, it is crucial for couples preparing for natural pregnancy to monitor sperm motility. OBJECTIVE This study verified the performance of an artificial intelligence–based image recognition and cloud computing sperm motility testing system (Bemaner, Createcare) composed of microscope and microfluidic modules and designed to adapt to different types of smartphones. METHODS Sperm videos were captured and uploaded to the cloud with an app. Analysis of sperm motility was performed by an artificial intelligence–based image recognition algorithm then results were displayed. According to the number of motile sperm in the vision field, 47 (deidentified) videos of sperm were scored using 6 grades (0-5) by a male-fertility expert with 10 years of experience. Pearson product-moment correlation was calculated between the grades and the results (concentration of total sperm, concentration of motile sperm, and motility percentage) computed by the system. RESULTS Good correlation was demonstrated between the grades and results computed by the system for concentration of total sperm (r=0.65, P<.001), concentration of motile sperm (r=0.84, P<.001), and motility percentage (r=0.90, P<.001). CONCLUSIONS This smartphone-based sperm motility test (Bemaner) accurately measures motility-related parameters and could potentially be applied toward the following fields: male infertility detection, sperm quality test during preparation for pregnancy, and infertility treatment monitoring. With frequent at-home testing, more data can be collected to help make clinical decisions and to conduct epidemiological research.

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Semen Production Characteristics of Pasundan Bull at Different Body Weight

Jurnal Agripet ◽

10.17969/agripet.v21i1.15186 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Anggit Damaratri Lapoliwa ◽

Nurul Isnaini

Keyword(s):

Body Weight ◽

Sperm Motility ◽

Recovery Rate ◽

Sperm Concentration ◽

The Body ◽

Production Traits ◽

Motile Sperm ◽

Frozen Semen ◽

Semen Volume ◽

Production Characteristics

ABSTRACT. Pasundan cattle is currently proposed as a potential livestock to support national meat self-sufficiency program. However, the information regarding their reproductive performance is still very limited. This study was conducted to evaluate the semen production characteristics of Pasundan bull at different body weight. A total of 178 semen samples which were collected from one Pasundan bull were used in this study. The semen collection was done for 31 months and during this period, the body weight of Pasundan bull was classified into 4 categories, namely 400 kg, 400 to 500 kg, 500 to 600 kg, and ≥600 kg. The results showed that overall mean semen volume, semen pH, sperm concentration, total sperm, individual sperm motility, total motile sperm, post-thawing sperm motility, recovery rate of sperm motility, and frozen semen production were 5.89 ml/ejaculate, 6.69, 1.04 billion/ml, 6.04 billion/ejaculate, 55.75%, 3.40 billion/ejaculate, 40.91%, 58.20%, and 265.11 doses/ejaculate. The difference in body weight significantly affect semen volume (P=0.001), semen pH (P=0.001), sperm concentration (P=0.043), total sperm (P=0.002), individual sperm motility (P0.001), total motile sperm (P0.001), and frozen semen production (P=0.004). There was no significant effect (P0.05) of body weight on post-thawing sperm motility and recovery rate of sperm motility. In conclusion, the semen production traits of Pasundan bull are improved with the increase in body weight up to 500 to 600 kg and remain stable at the body weight of ≥600 kg. (Karakteristik produksi semen sapi Pasundan pada bobot badan yang berbeda) ABSTRAK. Sapi Pasundan saat ini diajukan sebagai salah satu ternak potensial untuk mendukung program swasembada daging nasional. Akan tetapi, informasi tentang penampilan reproduksinya masih sangat terbatas. Penelitian ini dilakukan untuk mengevaluasi karakteristik produksi semen sapi pejantan Pasundan pada bobot badan yang berbeda. Sebanyak 178 sampel semen yang dikoleksi dari 1 ekor sapi pejantan Pasundan digunakan pada penelitian ini. Penampungan semen dilakukan selama 31 bulan dan pada periode tersebut, bobot badan sapi pejantan Pasundan diklasifikasikan menjadi 4 kategori yaitu 400 kg, 400 to 500 kg, 500 to 600 kg, and ≥600 kg. Hasil penelitian menunjukkan bahwa rata-rata volume semen, pH semen, konsentrasi sperma, total sperma, motilitas individu sperma, total sperma motil, motilitas sperma post-thawing, nilai recovery rate, dan produksi semen beku adalah 5,89 ml/ejakulat, 6,69, 1,04 miliar/ml, 6,04 miliar/ejakulat, 55,75%, 3,40 miliar/ejakulat, 40,91%, 58,20%, dan 265,11 dosis/ejakulat. Perbedaan bobot badan memberikan pengaruh terhadap volume semen (P=0,001), pH semen (P=0,001), konsentrasi sperma (P=0,043), total sperma (P=0,002), motilitas individu sperma (P0,001), total sperma motil (P0,001), dan produksi semen beku (P=0,004). Di sisi lain, bobot badan tidak memberikan pengaruh (P0,05) terhadap motilitas sperma post-thawing dan nilai recovery rate. Kesimpulannya, karakteristik produksi semen sapi pejantan Pasundan meningkat seiring dengan peningkatan bobot badan hingga 500 sampai 600 kg dan tetap stabil hingga ≥600 kg.

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Comparative analysis of various step-dilution techniques on the quality of frozen Limousin bull semen

Veterinary World ◽

10.14202/vetworld.2020.2422-2428 ◽

2020 ◽

Vol 13 (11) ◽

pp. 2422-2428

Author(s):

Ani Atul Arif ◽

Tulus Maulana ◽

Ekayanti Mulyawati Kaiin ◽

Bambang Purwantara ◽

Raden Iis Arifiantini ◽

...

Keyword(s):

Sperm Motility ◽

Artificial Insemination ◽

Skim Milk ◽

Egg Yolk ◽

Dna Integrity ◽

Dilution Technique ◽

Bull Semen ◽

Frozen Semen ◽

One Step

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.

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