• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.298-304
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• 2018

The single-vessel multienzyme UDP-${\alpha}$-$\text\tiny{D}$-glucose recycling system was coupled with a forward glucosylation reaction to produce novel glucose moiety-conjugated derivatives of different tetracycline antibiotic analogs. Among five tetracycline analogs used for the reaction, four molecules (chlorotetracycline, doxytetracycline, meclotetracycline, and minotetracycline) were accepted by a glycosyltransferase enzyme, YjiC, from Bacillus licheniformis to produce glucoside derivatives. However, the enzyme was unable to conjugate sugar units to rolitetracycline. All glucosides of tetracycline derivatives were characterized by ultraviolet absorbance maxima, ultra-pressure liquid chromatography coupled with photodiode array, and high-resolution quadruple time-of-flight electrospray mass spectrometry analyses. These synthesized glucosides are novel tetracycline derivatives.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.175-179
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• 2007

본 연구는 0.5ml straw를 이용한 정자 동결융해 시 응해 온도가 동결정자의 성상에 미치는 영향을 파악하고 미니 돼지의 동결정액에 최적화된 적정 응해 조건을 찾기 위하여 실시하였다. 정액동결 straw를 37, 50 및 $70^{\circ}C$에서 각각 5초, 10초 및 45초간 융해하여 생존율(SYBR-14/PI staining), 정자원형질막기능검사 (Hypoosmotic Swelling Test) 및 첨체반응율(CTC : chlorotetracycline staining)을 검사 한 결과, $70^{\circ}C$에서 5초간 융해한 정자의 생존율과 CTC 결과가 $37^{\circ}C$와 $50^{\circ}C$에서 10초 및 45초간 융해한 처리구보다 유의적(p<0.05)으로 높은 생존율과 낮은 비율의 첨체 반응율을 얻었다. 따라서 미니돼지의 동결 정액은 고온에서 단시간 융해를 하는 것이 정자의 성상에 유리한 것으로 사료된다.

• 한국수정란이식학회지
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• 제26권1호
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• pp.71-78
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• 2011

This study was undertaken to find out the effect of cholesterol and serum albumin on sperm ability and lipid peroxidation levels period to the liquid storage of miniature pig sperm. Ejaculated semen from miniature pigs was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle, and extended with Modena solution {with and without BSA, methyl-beta-cyclodextrin (-cholesterol) and cholesterol loaded cyclodextrin (+cholesterol)}. Each semen was assessed for viability (SYBR-14/PI staining) and acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining at 1, 3, 5, 7 and 10 days of storage. At for the effects of cholesterol and serum albumin on lipid peroxidation, semen were incubated with $H_2O_2$ ($10\;{\mu}M$), and lipid peroxidation level were measured by flow cytometry using the lipid peroxidation reporter probe $C_{11}-BODIPY^{581/591}$. The result, lipid peroxidation level in sperm added with cholesterol were lower in $10\;{\mu}M$ $H_2O_2$ compared to the added sperm with serum albumin. Also, added cholesterol to sperm had significant (p<0.05) higher viability when storage for 7 and 10 days and lower when 10 days of storage percentage of acrosome-reacted sperm (AR pattern) in acrosome state as say result compared to other treated groups. In conclusion, role of cholesterol during lipid storage in miniature pig spermatozoa was protected boar spermatozoa from lipid peroxidation prior to lipid storage. Addition serum albumin during lipid storage in sperm may be induce sperm membrane damage by lipid peroxidation. Therefore, addition of cholesterol to miniature pig sperm will be lead to extension of liquid storage periods.

• Asian-Australasian Journal of Animal Sciences
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• 제25권8호
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• pp.1153-1158
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• 2012

The objective of this study was to evaluate the antioxidant capacity of Lactobacillus plantarum ZLP001 and its effects on growth performance and antioxidant status in weaning piglets. The survival in hydrogen peroxide and free radical-scavenging activity of Lactobacillus plantarum ZLP001 were analysed in vitro. The Lactobacillus plantarum ZLP001 showed high viability in 1.0 mmol/L hydrogen peroxide and high scavenging ability against hydroxyl, superoxide anion, and DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals which was dose dependent. Ninety-six weaning piglets were selected ($7.45{\pm}0.79kg$) and divided into three groups comprising of negative control without any supplementation, treatment group with supplemented $6.8{\times}10^7$ Lactobacillus plantarum ZLP001 CFU/g of diet, and positive control with antibiotic treatment (chlorotetracycline, 80 mg/kg diet). The results showed that Lactobacillus plantarum ZLP001 supplementation enhanced feed conversion rates in piglets compared with control (p<0.05). Supplementation of Lactobacillus plantarum ZLP001 increased the concentration of superoxide dismutase (p<0.05), glutathione peroxidase (p<0.01) and catalase in serum (p<0.10), while decreased the concentration of malondialdehyde (p<0.05). The present study implies that the strain Lactobacillus plantarum ZLP001 had high antioxidant ability and its supplementation improved the growth performance and antioxidant status of weaning piglets, so it can be considered useful to alleviate oxidative stress and increase productive performance of pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.351-358
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• 2007

Carbohydrate-protein interactions are known to be important in gamete interactions. Several evidence indicated that a fucose-containing sulfated polysaccharide fucoidan was potential inhibitor of fertilization in vitro and thus fucose seemed to be part of the recognition signal of gamete interaction in mammals. In recent investigation we found that ${\alpha}$-L-fucosidase activity was present in boar spermatozoa and it was related to sperm binding to and penetration into zona pellucida (ZP) in vitro. The objective of this study was to determine the effects of monosaccharide L-fucose and polysaccharide fucoidan on sperm ${\alpha}$-L-fucosidase activity and relation to sperm-oocyte interaction in pig. Results indicated that the activity of sperm ${\alpha}$-L-fucosidase was largely inhibited (62%) when sperm suspension was treated with monosaccharide L-fucose. It also significantly inhibited the number of sperm binding to ZP (32%) and penetration into zona-intact oocytes (72%), but did not inhibit penetration into zona-free oocytes when fertilization medium contained L-fucose. The chlorotetracycline (CTC) assessment showed that L-fucose did not affect induction of sperm capacitation and acrosome reaction. In contrast, the activity of sperm ${\alpha}$-L-fucosidase was not inhibited when sperm suspension was treated with polysaccharide fucoidan but sperm-ZP binding was greatly inhibited (85%) and completely blocked sperm penetration into zona-intact or zona-free oocytes. The CTC assessment showed that fucoidan increased the F pattern and decreased the AR pattern sperm. These results suggested that the different inhibitory mechanisms were present between monosaccharide L-fucose and polysaccharide fucoidan on sperm-oocyte interaction, the inhibition effect of ${\alpha}$-L-fucose on sperm binding and penetrating into ZP caused sperm ${\alpha}$-L-fucosidase inhibited by ${\alpha}$-L-fucose.

• Reproductive and Developmental Biology
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• 제29권4호
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• pp.247-252
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• 2005

The purpose of this study was to assess sperm quality during in vitro storage of miniature-pig semen in order to determine which extender should be used and how extender can be diluted for in vitro storage of miniature-pig semen. Freshly ejaculated miniature-pig's semen was diluted with same volumes of Beltsville Thawing Solution (BTS), Androhep, Modena, Mulberry III and modified-Modena extenders. Sperm quality was evaluated by examining viability, motility, abnormality, acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining. Sperm motility decreased with storage period prolonged and differences among BTS, Androhep, Modena and Mulberry III were apparent On Day 1, approximately 80% of the sperm were motile, but motility decreased to $40\%$ at Day 7. During the 7 days of storage, sperm survival in modified-Modena B extender was higher than another extenders. However, it was not differ significantly among other extenders. The percentage of F and B patterns were not differ significantly among the extenders. However, F pattern in modified-Modena B extender was slightly higher until 3 days of storage than that of Modena extender, modified-Modena A extender and modified-Modena C extender. The percentage of AR patterns in modified-Modena B extender was slightly lower, but did not differ significantly among other extenders. The results of present study suggest that modified-Modena B was effective as new extender for in vitro storage of miniature-pig semen.

• 한국수산과학회지
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• 제38권6호
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• pp.372-378
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• 2005

An analysis method of tetracyclines far fish and shellfish products containing large amount of low molecular materials and pigments was established. The recovery of the established analysis method for four tetracycline samples was $72-100\%$ and higher than other methods reported. Especially, proposed sample treatment protocol was shown to be effective for the removal of low molecular materials and pigments that tend to interfere with accurate analysis. The detection limit of oxytetracycline (OTC) and tetracycline (TC) from the sample was 0.02 ppm, and the detection limit of chlortetracycline (CTC) and doxycycline (DC) from the sample was 0.1ppm, To examine the efficiency of the established method and identify tetracycline usage in fish farms, tetracycline group antibiotics in the flounder being cultured was monitored. The improved method can be used for fish and shellfish products effectively and all surveyed fish farms have used tetracycline all the year round. The proposed method was adopted as official method for fishery products by Korean Food and Drug Administration in 2003 and it is being used by regulatory authority as National Fishery Products Quality Inspection Service.

• 한국가금학회지
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• 제31권4호
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• pp.245-253
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• 2004

본 연구는 육계사료 내 rPST-yeast culture의 첨가 급여가 육계의 성장성적, 도체 특성 및 골격발달 등에 미치는 영향을 규명하기 위해 수행하였다. 2일령의 Ross 육용종 수평아리 460수를 공시하여 항생제, rPST-yeast culture 및 yeast culture가 첨가되지 않은 대조구 사료와 대조구 사료에 항생제(chlorotetracycline)만을 $0.1\%$ 수준으로 첨가한 실험사료(T1), rPST-yeast culture를 각각 $0.1\%$ 및 $0.2\%$ 수준으로 첨가한 실험사료(T2 및 T3) 및 yeast culture만을 $0.2\%$수준으로 첨가한 실험사료(T4)를 각각 6주간 급여하였다. 사료섭취량과 증체량은 주 단위로 조사하였다. 실험 5주차 종료 시에 각 처리구별로 10수씩 선발하여 도살하였고, 간, 비장, 복강지방 및 가슴근육의 상대적 중량과 가슴근육의 화학적 조성을 측정하였다. 혈액 내 GOT와 GPT 수치, 총 콜레스테롤, Ca 및 P을 측정하였으며, 경골의 중량, 파쇄 강도, 회분 함량, Ca및 P 함량을 측정하였다. 증체량은 대조구에 비하여 rPST-yeast culture를 $0.1\%$ 첨가한 실험구에서 증가하는 경향을 보였으며, 사료섭취 량 및 사료요구율은 처리구간 큰 차이가 없었다. 비장 및 복강 지방의 상대적 중량은 실험구간 유의한 차이가 관찰되지 않았으나, 간과 가슴근육의 상대적 중량은 처리간에 유의한 차이가 관찰되었는데,특히 T2처리구에서 가슴근육의 상대적 중량이 대조구에 비해 유의하게 증가하는 결과가 관찰되었다(p<0.05). 가슴 근육 내 조단백질 및 조지방 함량에 처리구간에 큰 차이는 없었으나, 수분 함량은 대조구에 비해 rPST-yeast culture와 SC yeast culture 첨가 급여구에서 유의하게 증가하였다(P<0.05). 혈청 내 GOT 수치는 변화가 없었으나, GPT수치는 rPST-yeast culture 첨가 급여 후에 유의하게 감소하는 결과가 얻어졌다(P<0.05). 혈중 총 콜레스테롤, Ca 및 P 농도는 처리간에 큰 차이를 보이지 않았다. 경골중량, 파쇄강도 및 화학적 성분 조성에서도 처리간에 큰 차이는 나타나지 않았다. 본 연구의 결과로부터 사료 내 rPST-yeast culture의 첨가급여에 의해 육계에서 성장성적이 향상될 가능성이 제시되었고, 특히 가슴근육이 증가와 같은 긍정적인 효과를 기대할 수 있을 것으로 사료되었다.

• Journal of Animal Science and Technology
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• 제44권3호
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• pp.289-296
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• 2002

본 시험은 yeast culture제제들(Saccharomyces cerevisiae, Pichia pastoris)의 첨가가 육계의 성장, 장내 미생물균총 및 혈청 IgG농도에 미치는 영향을 측정하기 위하여 기초사료(control)에 chlorotetracycline 100ppm(CTC), Saccharomyces cerevisiae 0.3%(YC-SC), Pichia pastoris 0.3% (YC-PP), 정제된 Pichia pastoris culture 0.1% (RPPC-0.1) 및 0.3%(RPPC-0.3)를 각각 첨가하였다. Ross 종 육계 1000수(암․수 각각 500수)를 20개의 floor pen에 50수(암 수 각 25수)씩 수용하여 control, CTC, YC-SC와 RPPC-0.1구는 3반복으로 YC-PP와 RPPC-0.3구는 4반복으로 배치하였다. 증체량은 처리간에 유의한 차이가 없었으나 대조구나 항생제첨가구에 비해 효모배양물의 첨가구(YC-SC, RPPC-0.1 및 RPPC- 0.3)에서 증체량이 약 4-5% 개선되는 효과가 있었다. 항생제와 효모배양물은 첨가한 구가 대조구에 비해 사료효율이 개선되며 폐사율이 감소하는 경향이 있었다. 영양소 이용율은 yeast culture 첨가에 의해 유의한 영향을 받지 않았다. 소장하부 내용물의 Lactobacilli의 수는 효모배양물의 첨가구(YC-SC, RPPC-0.1 및 RPPC-0.3)에서 대조구보다 다소 높은 경향을 보였고, Cl. perfringens의 수는 CTC와 정제된 Pichia pastoris culture의 첨가에 의해 뚜렷이 저하되었다. E. coli의 수는 정제된 Pichia pastoris 구들이 낮았는데 특히 RPPC-0.3가 대조구보다 유의하게(P$\prec$0.02) 낮았다. 혈청 IgG 농도는 처간에 유의한 차이는 없었으나 효모배양물의 첨가구들에서 control구나 항생제 첨구에서보다 높은 경향을 나타내었다. 결론적으로 본시험에 사용된 yeast culture제제는 생산성, 소장내 미생물 균총과 혈청 IgG에 있어서 유의한 차이는 없었으나 항생제(CTC)와 같거나 그 이상의 개선효과를 나타내었다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.385-390
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• 2011

The purpose of this study was undertaken to evaluate of cryopreservation efficiency in ${\alpha}$ 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. After thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.