• Asian-Australasian Journal of Animal Sciences
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• 제5권3호
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• pp.449-454
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the incidence of the morphologically abnormal spermatozoa. Semen was collected twice a week by A. V. over one-year period. The percentage of total abnormal spermatozoa was $14.1{\pm}0.5$. Ejaculates collected during hot summer season had significantly higher incidence of abnormal spermatozoa than those collected during winter time. Warm spring had moderate semen abnormality. In addition to its effect on the total number of morphologically abnormal spermatozoa, season affected significantly the primary as well as the secondary types of abnormalities. The differences between incidence of primary and secondary types of abnormalities were not significant for all seasons and seasons pooled together. The ratio between the total forms of abnormality in the head, mid-piece and tail of spermatozoa was as 1:1.5:1. Head and mid-piece had defected more during summer compared with both winter and spring. There was no variation in tail abnormalities due to season. The significant effect of season on head was observed by large, pyriform, free and detached heads, while that on mid-piece was by swollen, coiled mid-pieces and protoplasmic droplets.

• Reproductive and Developmental Biology
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• 제29권4호
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• pp.235-239
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• 2005

This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

• 한국동물생명공학회지
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• 제36권2호
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• pp.82-90
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• 2021

In the present study, we examined if deep uterine artificial insemination (DUAI) can improve the pregnancy rate of artificial insemination (AI) using epididymal spermatozoa (ES) in Hanwoo cattle. The estrus cycles of 88 Hanwoo cows were synchronized, and 17 cows were artificially inseminated using the DUAI method with ES, 20 cows were artificially inseminated via the uterine body (BUAI) method with ES, and as a control, 51 cows were inseminated by using the BUAI method with ejaculated spermatozoa from 1 proven bull after frozen thawing. The pregnancy rate of the DUAI method (58.8%) was higher than that of the BUAI method (25.0%, p = 0.0498). The motility of ES was examined immediately after thawing and after 3 and 6 h of incubation. The rapid progressive sperm motility of the control group was significantly higher than that of the ES group immediately after thawing and after 3 and 6 h of incubation (p < 0.05). The straight line velocity and average path velocity of the ES group after 6 h of incubation were significantly lower than those in the control group (p < 0.05). The linearity and amplitude of lateral head of ES were lower than those at 6 h (p < 0.05). The flagellar beat cross frequency and hyperactivation of ES were lower than the control spermatozoa immediately after thawing and at 3 h (p < 0.05). These motility parameters suggested that ES had a low motility and fertilization ability compared to the control spermatozoa. After frozen-thawing and 3 h of incubation, the percentage of live spermatozoa with intact acrosomes in the ES was significantly lower than that in ejaculated spermatozoa (p < 0.05). Our findings suggested that the DUAI method can overcome the low pregnancy rate of ES, despite the low motility, viability, and fertilization ability of ES.

• Asian-Australasian Journal of Animal Sciences
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• 제5권3호
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• pp.439-447
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the biophysical characteristics of semen. Semen was collected twice a week by AV (artificial vagina) over one-year period. The analyses revealed that all the basic seminal traits studied were differed significantly due to season, except the ejaculate volume and consistency and the percentage of swollen spermatozoa in a hypo-osmotic fructose-citrate solution. Ejaculates collected during hot summer season had significantly lower sperm motility, concentration and total counts, and higher percentage of dead spermatozoa than those collected during winter time. Warm spring had moderate semen quality. The temperature-humidity index was calculated and it was associated (p < 0.01) negatively with the ejaculate pH, sperm concentration and total counts, and positively with the % of dead sperms. Ejaculate volume, percentage of swollen spermatozoa, individual motilities did not correlate significantly with the change in temperature-humidity index values. The total live, motile spermatozoa per ejaculate during both the winter and spring seasons showed significant increase of about 37% and 32% respectively over the summer season. Also, rectal temperatures of the bulls were elevated during the hot summer season, while the values of blood hemoglobin and packed-cell volume were decreased.

• Asian-Australasian Journal of Animal Sciences
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• 제13권7호
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• pp.901-905
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• 2000

An experiment was conducted to investigate the effect of caffeine, cAMP and cattle seminal plasma on preservation of semen at ultra low temperature ($-196{^{\circ}C}$). Each semen sample was divided into four parts equal in volume and sperm concentration; three were treated with caffeine, or cAMP, or cattle seminal plasma (CSP) and the fourth was kept as control. Sperm motility, abnormal spermatozoa, live-dead count and acrosomal damage were studied at different stages of freeze preservation viz.; just after dilution, at $5{^{\circ}C}$, at glycerolisation, before freezing, just after freezing, 24 hours of storage, and one week of storage. Sperm motility (58.39, 61.33, 52.00 and 50.39 per cent), non-eosinophilic spermatozoa (72.55, 69.98, 63.31 and 67.64 per cent), abnormal spermatozoa (5.71, 4.98, 8.04 and 5.66 per cent) and acrosomal damage (13.28, 13.33, 14.80 and 14.65 per cent) were observed in cAMP, caffeine, cattle seminal plasma and control, respectively, at every stage of freeze preservation. From this study it could be concluded that freezability of buffalo semen can be improved through the addition of caffeine followed by cAMP and cattle seminal plasma.

• Asian-Australasian Journal of Animal Sciences
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• 제23권7호
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• pp.873-879
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• 2010

We investigated the effect of various concentrations of taurine during in vitro fertilization (IVF) on the embryonic development up to the blastocyst stage of bovine oocytes fertilized with three different Japanese Black bulls (Bull A, B and C). In vitro matured oocytes were fertilized with various concentrations of taurine (0, 1, 10, 50 and 100 mM) in the presence of 2.5 or 5.0 mM caffeine plus $25{\mu}g$/ml heparin (CH) for 6 hr or $100{\mu}g$/ml heparin (H) for $24{\pm}2$ h. After IVF, the cleavage rates from the 2 to 16 cell stage determined at 3 days and the development rates up to the blastocyst stage determined at 7-8 days from the onset of IVF were assessed. Although the cleavage rates for the taurine concentration groups were not significantly increased in any of the three bulls in the CH groups, the development rates up to the blastocyst stage of the 50 mM taurine group of Bulls A and B, and of the 1 to 50 mM groups of Bull C were increased (p<0.05) compared to those of the control (0 mM taurine) groups. On the other hand, none of the bulls in the H groups showed any significant increase either in the cleavage rates or blastocyst formation rates in any taurine concentrations groups compared with those of the control groups. These results indicate that the addition of 50 mM taurine to a fertilization medium containing caffeine and heparin may stimulate embryonic development up to the blastocyst stage when fertilized with different bull semen.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.295-300
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• 2011

The objective of this study was to evaluate efficiency of methyl-beta-cyclodextrin (MBCD) in the sperm preservation of bull. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 1, 5, 10 and 20 mM MBCD before freezing process. Analysis of viability in frozen-thawed sperm was estimated by SYBR14/PI double stain, hypoosmotic swelling test(HOST) and acrosome damage with FITC-PNA, and mitochondria activation with Rhodamin123 by flow-cytometry. The sperm viability was significantly in 0 mM and 5 mM concentrations of MBCD than other groups (p<0.05). However, the HOST was significantly lower at 20 mM concentration of MBCD than other concentrations (p<0.05). In addition, acrosome damage and mitochondria activation rates were significantly lower at 20 mM concentration of MBCD than other groups (p<0.05). In conclusion, the viability of sperm was not significantly different among concentrations of MBCD 0, 5 and 10 mM, but MBCD 20 mM was significantly lower than other groups. In addition, as concentrations of MBCD was high, HOST, acrosome damage and mitochondria activation rates had a negative effect in bull sperm.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1411-1420
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• 2020

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.

• Animal Bioscience
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• 제36권2호
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• pp.209-217
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• 2023

Objective: The conservation of Bali bulls, the Indonesian native breed of cattle, is crucial for cattle breeding in Indonesia. To guarantee the spread of Bali bulls through artificial insemination the quality of the frozen semen must be high. To this end, adding an extender material to semen that increases spermatozoa's survival during cryopreservation is important. Green tea extract (GTE) can be used as cryoprotectant because its high antioxidant activity can help avoid reactive oxygen species formation. Methods: Semen of five Bali bulls from the National Artificial Insemination Center at Singosari, Indonesia was collected routinely twice a week. First, fresh semen inspection was performed to determine the feasibility of using Bali bulls as animal samples. The extender used in this study was Tris-based egg yolk. The samples were divided into four treatments: T0, no GTE added to the extender; T1, 0.05 mg GTE plus 100 mL extender; T2, 0.10 mg GTE plus 100 mL extender; and T3, 0.15 mg GTE plus 100 mL extender. The semen freezing process was conducted according to standard procedures and sperm quality parameters, i.e., sperm motility, viability, abnormalities, and membrane integrity observed pre-freezing and post-thawing. Results: There were significant differences in total motility, progressive motility, viability, and integrity membrane of Bali bull sperm at both pre-freezing and post-thawing after adding GTE into the extender. In contrast, there were no differences in abnormalities among treatments. Conclusion: Adding GTE at a 0.15 mg into 100 mL Tris-based egg yolk extender can improve the quality of cryopreserved Bali bull sperm.

• 한국가축번식학회지
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• 제12권2호
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• pp.112-119
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• 1988

These studies were conducted to investigate the effects of culture temperature and time on the in vitro maturation and semen type and media on the in vitro fertilization of bovine follicular oocytes, and to asses in vitro fertilization rate of oocytes cultured by extraffollicular method following fertilization in vitro, or transfer into the pseudopregnant rabbit oviduct or uterus. The bovine oocytes recovered from follicles were cultrued for 18 hrs or 72hrs at 38$^{\circ}C$ with 5% CO2 in moist air. Flesh-diluted(2 folds) and frozen-thawed semen in 0.5ml straw from a fertile bull were used. In order to obtain capacitation of spermatozoa were treated with bovine follicular fluids(BFF) and Inophore A(IA). The results obtained were summarized as follows: 1. The oocytes were classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 62.0%, 12.0%, 17.2%, 5.9% and 3.0% of the total oocytes harvested, respectively. 2. The oocytes matured to metaphase II were significantly increased between 24-48hrs of incubation and at 37-39$^{\circ}C$ with 5% CO2 in moist air. 3. The in vitro fertilization rate following transferred into rabbit oviduct or uterus with bull semen and in vitro matured oocytes were higher ligation than non-ligation of oviduct or uterus. 4. The in vitro fertilization rate of oocytes matured in vitro were higher neat than frozen semen and treatment of IA than BFF on the capacitation of spermatozoa. 5. The effects of semen types and media on in vitro fertilization of oocytes matured in vitro were higher fertilization rate of neat than friozen semen, and media was not significant.