• Title/Summary/Keyword: VapA

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Effect of Optixcell and Triladyl extenders on frozen-thawed sperm motilities and calving rates following artificial insemination in Hanwoo

  • Kang, Sung-Sik;Lee, Myung-Suk;Kim, Ui-Hyung;Lee, Seok-Dong;Yang, Byoung-Chul;Yang, Boh-Suk;Cho, Sang-Rae
    • Korean Journal of Agricultural Science
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    • v.46 no.1
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    • pp.195-204
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    • 2019
  • In this study, we examined the effect of a liposome-based extender (Optixcell) and a tris-citric egg-yolk extender (Triladyl) on the frozen-thawed spermatozoa characteristics and the calving rate. The percentages for the total motility of the frozen-thawed spermatozoa were similar in the Optixcell and Triladyl groups. However, among the motile spermatozoa with a straight line velocity (VSL) ${\geq}25{\mu}m/sec$, the curvilinear velocity (VCL, ${\mu}m/sec$), VSL (${\mu}m/sec$), average path velocity (VAP, ${\mu}m/sec$), amplitude of lateral head displacement (ALH, ${\mu}m$), beat cross frequency (BCF, Hz), and plasma membrane integrity of the frozen-thawed spermatozoa for the Optixcell group were significantly higher than those for the Triladyl group. Furthermore, the calving rate in the Optixcell group (79.0%) was higher than that of the Triladyl group (62.8%). However, the acrosomal membrane integrity of the frozen-thawed spermatozoa in the Optixcell and Triladyl groups was not significantly different. These results indicate that semen freezing with Optixcell improved the motility and plasma membrane integrity of frozen-thawed spermatozoa and the calving rate of Hanwoo cows (native Korean cattle). In conclusion, our results suggest that semen freezing with the liposome-based extender Optixcell is more efficient than with the tris-citric egg-yolk extender Triladyl for improved offspring production.

Disease monitoring of cultured rainbow trout and coho salmon in Gangwon province in 2021 (2021년 강원도 양식 무지개송어 및 은연어 비법정전염병 모니터링)

  • Soo-ji, Woo;Seung Hoon, Lee;So-Sun, Kim;Soon-Gyu, Byun;Joon-Young, Song;Seong Don, Hwang
    • Journal of fish pathology
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    • v.35 no.2
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    • pp.215-223
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    • 2022
  • Disease including parasite, bacteria and virus cause serious mortality to salmonid fish in the aquaculture. In this study, we investigated the current disease status of the rainbow trout (Oncorhynchus mykiss) and coho salmon (Oncorhynchus kisutch) in Yanayang, Pyeongchang, Jeongseon and Yeongwol of Gangwon province in 2021 and performed molecular characterization of those pathogen. For parasites, Ichthyophthirius multifiliis was observed at 2 farms. For bacteria, we identified Aeromonas sobria from kidney of rainbow trout using phylogenetic analysis of gyrB gene. A. salmonicida were isolated from necrosis site of gill cover and fin in coho salmon and necrotic lesion of fin in rainbow trout. Phylogenetic analysis using vap gene indicated that A. salmonicida isolated in this study were clustered with previously reported A. salmonicida subsp. salmonicida isolates. For virus, JRt-Nagano type of infectious haematopoietic necrosis virus was detected in rainbow trout, but infectious pancreatic necrosis virus and Oncorhynchus masou virus were not detected. These results provide useful information for the prevention of disease spread and transmission when cultivating new species such as Atlantic salmon in Korea.

Analysis of Semen Parameters in α1,3-Galactosyltransferase-/- Boars

  • Hwang, In-Sul;Lee, Seung-Chan;Kim, Sung Woo;Kwon, Dae-Jin;Park, Mi-Ryung;Yang, Hyeon;Oh, Keon Bong;Ock, Sun-A;Woo, Jae-Seok;Im, Gi-Sun;Hwang, Seongsoo
    • Journal of Embryo Transfer
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    • v.32 no.2
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    • pp.53-58
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    • 2017
  • It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous ${\alpha}1,3$-Galactosyltransferase knock-out ($GalT^{-/-}$) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 $GalT^{-/-}$ boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.

Association Study Analysis of Phospholipase C Zeta (PLCz) Gene Polymorphism (g.158T>C) for Duroc Boar Post-Thawed Semen Motility and Kinematic Characteristics (PLCz 유전자의 유전적 다형성(g.158T>C)과 두록 동결정액의 운동학적 특성과의 연관성 분석)

  • Sa, Soo-Jin;Lee, Mi-Jin;Kim, Ki-Hyun;Woo, Jae-Seok;Ko, Jun-Ho;Kim, Young-Ju;Cho, Eun-Seok
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.137-142
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    • 2015
  • Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.158T>C) of phospholipase C zeta (PLCz) gene reported to be significant association with MOT. This study was conducted to evaluate the PLCz gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.158 T>C SNP of PLCz was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.0001, p=0.0002, p<0.0001 and p<0.0001, respectively). Therefore, we suggest that the intron region of the porcine PLCz, may be used as a molecular marker for Duroc boar post-thawed semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

Association with Post-Thawed Semen Motility and Kinematic Characteristics of g.35756 T>C on Estrogen Receptor 1 (ESR1) Gene in Duroc Pigs (두록의 동결정액의 운동학적 특성과 ESR1 유전자의 SNP(g.35756T>C)와 연관성 분석)

  • Cho, Eun-Seok;Kim, Ki-Hyun;Woo, Jae-Seok;Lee, Mi-Jin;Ko, Jun-Ho;Kim, Young-Ju;Sa, Soo-Jin
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.143-147
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    • 2015
  • Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen Post-thawed in boar. Recently, polymorphism (g. 35756 T>C) of Estrogen Receptor 1 (ESR1) gene reported to be significant association with MOT. This study was conducted to evaluate the ESR1 gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.35756 T>C SNP of ESR1 was significantly associated with frozen semen motility and kinematic characteristics. The g.35756 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.001). The SNP was also significantly associated with ALH (P<0.05). Therefore, we suggest that the g. 35756 T>C polymorphism in the intron 1 region of the porcine ESR1 gene could potentially be applied in frozen semen programs to improve MOT trait, but only after validation in other populations.

Primary Culture of Chicken Tracheal Epithelial Cells and Study on Those Characters for Recombinant Virus Infection (닭 기도 상피세포의 1차 배양과 유전자 재조합 바이러스의 감염 특성 연구)

  • Park, Mi Na;Jang, Hyun-Jun;Keum, Dae Ho;Choi, Jin Ae;Yoo, Jae Gyu;Byun, Sung June;Park, Jong Ju;Ji, Ju Young;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Korean Journal of Poultry Science
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    • v.40 no.4
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    • pp.299-304
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    • 2013
  • Tracheal epithelial cells (TECs) are an important tool for studies of viral respiratory diseases. Primary TECs have been cultured from human, mouse and hamster. It is also necessary to diagnose viral respiratory disease and reveal infection mechanisms in chicken. In this study, we isolated tracheal epithelial layers from tracheal of 20-day-old chicks and cultured primary TECs from the isolated layers. Ciliated cells which were a typical morphology of TECs were observed in cultured primary TECs and maintained until cell passage 5 (15 to 20 days). When we analyzed expression patterns of epithelial marker genes (retinoic acid responder, FGF-binding protein, virus activating protease (VAP) in TECs compared to immortalized chicken embryonic fibroblast cell line (DF-1), all the marker genes are highly expressed in TECs than in DF-1. When TECs were cultured with 0.1 and 1 MOI of ND virus (rNDV-GFP strain) to test the susceptibility of TECs for ND virus, 12.6% and 48.2% of the incubated TECs were infected respectively. In addition, when DF-1 was incubated with 1 MOI of ND virus, the virus infection rate of DF-1 was three times lower than the virus infection rate of TECs. These data could contribute to study infection mechanisms of viral respiratory diseases and control them in chicken.

Isolation of High Yielding Alkaline Protease Mutants of Vibrio metschnikovii Strain RH530 and Detergency Properties of Enzyme

  • Chung, So-Sun;Shin, Yong-Uk;Kim, Hee-Jin;Jin, Ghee-Hong;Rho, Hyune-Mo;Lee, Hyune-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.349-354
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    • 2000
  • Abstract A facultative alkalophilic gram-negative Vibrio metschnikovii strain RH530, isolated from the wastewater, produced several alkaline proteases (VAP) including six alkaline serine proteases and a metalloprotease. From this strain, high yielding YAP mutants were isolated by NTG treatment. The isolated mutant KS1 showed nine times more activity than the wild-type after optimization of the culture media. The production was regulated by catabolite repression when glucose was added to the medium. The effects of several organic nitrogen sources on the production of the YAP were investigated to avoid catabolite repression. The combination of 4% wheat gluten meal (WGM), 1.5% cotton seed flour (eSF), and 5% soybean meal (SBM) resulted in the best production when supplemented with 1% NaCl. The YAP showed a resistance to surfactants such as $sodium-{\alpha}-olefin$ sulfonate (AOS), polyoxy ethylene oxide (POE), and sodium dodecyl sulfate (SDS), yet not to linear alkylbenzene sulfonate (LAS). However, the activity of the YAP was restored completely when incubated with LAS in the presence of POE or $Na_2SO_4$. The YAP was stable in a liquid laundry detergent containing 6.6% SLES (sodium lauryl ether sulfate), 6.6% LAS, 19.8% POE, and stabilizing agents for more than two weeks at $40^{\circ}C$, but the stability was sharply decreased even after 1 day when incubated at $60^{\circ}C$. A washing performance test with the YAP exhibited it to be a good washing power by showing 51 % and 60% activity at $25^{\circ}C{\;}and{\;}40^{\circ}C$, respectively, thereby indicating that the YAP also has a good detergency at a low temperature. All the results suggest that the YAP produced from the mutant strain KSI has suitable properties for use in laundry detergents.rgents.

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Association study analysis of CD9 as candidate gene for Duroc pig sperm motility and kinematic characteristics (두록 정자 운동학적 특성과 후보유전자 CD9 유전자와의 연관성 분석)

  • Jeong, Yong-dae;Jeong, Jin-Young;Kim, Ki-Hyun;Cho, Eun-Seok;Yu, Dong-Jo;Choi, Jung-Woo;Jang, Hyun-Jun;Park, Sungk-won;Sa, Soo-Jin;Woo, Jae-Seok
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.281-285
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    • 2016
  • Cluster-of-differentiation antigen 9 (CD9) gene expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as a candidate gene to investigate Duroc boar semen motility and kinematic characteristics. This study was performed to investigatetheir association with semen motility and kinematic characteristics. DNA samples from 96 Duroc pigs with records of sperm motility and kinematic characteristics [Total motile spermatozoa (MOT, $82.27{\pm}5.58$), Curvilinear velocity(VCL, $68.37{\pm}14.58$), Straight-line velocity(VSL, $29.06{\pm}6.58$), the ratio between VSL and VCL(LIN, $47.36{\pm}8.42$), Amplitude of Lateral Head displacement(ALH, $2.88{\pm}0.70$)] were used in present study. A single nucleotide polymorphism (g.358A>T) in intron 6 was associated with MOT, VCL, VAP and ALH in Duroc population (p<0.05). Therefore, we suggest that the porcine CD9 may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not clear yet. These results will improve the understanding of the functions of the CD9 in spermatogenesis within the reproductive tracts, and will shed light on CD9 as a candidate gene in the selection of good sperm quality boars.

Effect of Herbal Extracts Mixtures on Antioxidant System in Chronic Enthanol-treated Rats

  • Kim, Mok-Kyung;Won, Eun-Kyung;Choung, Se-Young
    • Biomolecules & Therapeutics
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    • v.14 no.4
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    • pp.226-234
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    • 2006
  • Disturbance of antioxidant system is very common in chronic alcoholics and herbal or natural products with antioxidant activity have been used for its treatment. This study was to investigate the effect of Vitis vinifera extract(V), Schisandra chinensis extract(S), Taraxacum officinale extract(T), Gardenia jasminoides extract(G), Angelica acutiloba extract(A) and Paeonia japonica extract(P), and their combinations on the antioxidant and ethanol oxidation system. Male Sprague-Dawley rats were subjected to Lieber-DeCarli ethanol liquid diet(ED) and were then given different herbal extract mixtures for 6 weeks including VST(V 100+S 150+T 150mg/kg/day), VSG(V 100+S 150+G 150mg/kg/day), VTG(V 100+T 150+G 150mg/kg/day), and VAP(V 100+A 150+P 150mg/kg/day). When the activity of alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) were compared between ED only group and herbal extracts treatment group, the differences were statistically significant. Phase I and II(glutathione-S-transferase, phenol sulfatransferase) enzyme activities were found to be significantly higher in the VAT treatment group compared to the ED group. Herbal extracts not only repressed the ethanol-induced elevation of malondialdehyde level, but also protected against ethanol-induced decrease in glutathione content, glutathione reductase, glutathione peroxidase, catalase and superoxide dismutase activities. The administration of the herbal extracts was found to be effective in eliminating lipid-peroxides induced by long-term consumption of alcohol by activating various enzyme systems and physiological active compound formation system. After a chronic consumption of alcohol, Angelica Radix protected the liver via activating the ethanol-metabolism enzyme system, and Paeoniae Radix via activating the ethanol-metabolism enzyme and the phase I, II-metabolism enzyme system. Taraxaci Herba was also effective in liver protection via activating the ethanol-metabolism enzyme system and the phase I, II-metabolism enzyme system, Gardeniae Fructus via activating the phase II-metabolism enzyme system and the anti-oxidation system enzyme, and Schisandra Fructus and a grapestone via activating the anti-oxidation system. Our data suggest that these herbal extracts may be useful as a health functional food or new drug candidate for fatty liver and hepatotoxicity induced by chronic alcohol consumption.

Motional kinematics of Frozen-thawed Korean native cattle semen use of computer aided semen analysis(CASA) system (컴퓨터 정액자동분석에 의한 동결융해 한우 정액의 운동특성 연구)

  • Lee, Kang-nam;Lee, Byeong-chun;Kim, Jung-tae;Park, Jong-im;Shin, Tae-young;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.898-908
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    • 1998
  • The aim of this experiments were to assess the time-interval change of motional characteristics in frozen-thawed semen of Korean native cattle (KNC) by using computer aided semen analysis (CASA) technology. Twenty-six KNC frozen semen straws were obtained from Korean KNC improvement department, livestock improvement main division, national livestock cooperatives federation in Korea. Specimens were allowed to thaw at $37^{\circ}C$ for 30 sec in water bath. Semen analysis was performed on semen image analysis system (SIAS, Medical supply, Korea) adjusted to the gate settings and used the semen droplet ($5{\mu}l$) placed on Makler counting chamber (Sefi medical instrument, Israel) prewarmed at $37^{\circ}C$. The same person used the same micropipette to fill the Makler counting chamber. A total of 150 or more of sperms were analysed in each specimen by a single trained person by scanning at least 5 to 10 fields. The measurement parameters in SIAS were as follows ; frame rate = 30 frames per sec, image capture = 1 sec, minimum motile speed = $10{\mu}m/s$, maximum countable sperm number = 400. Statistical analysis was done by Student t-test with use of the Sigma plot program on a IBM personal computer. The dancemean(DNM) and hyperactivated sperm(HYP) of frozen-thawed KNC semen kinematics were significantly decreased(p < 0.05) after 10 min of incubation at $37^{\circ}C$ water bath. But, wobble(WOB) of same sample semen was significantly increased(p < 0.05) after 10 min of incubation and significantly decrease(p < 0.05) after 60 min of same incubation. And, after 30 mim of incubation, significantly differences were found most of motion kinematics, motifity(MOT), curvilinear velocity(VCL), straight line velocity(VSL), average path velocity(VAP), amplitude of lateral head displacement(ALH), beat cross frequency(BCF), mean angular displacement(MAD), dance(DNC), on same sample semen. The DNM of KNC semen sample was variable kinematics after 30 min of incubation. Also, the linearity(LIN) and straightness(STR) was significantly decreased(p < 0.05) from 60 min of incubation. In conclusion, the AI within 30 min after thawing of frozen semen can be an effective method for obtaining high fertility rate in KNC reproductive program.

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