• Title/Summary/Keyword: motile cells

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Isolation of Bacillus amyloliquefaciens ATC6 Producing Acidic Cellulase (산성 Cellulase를 분비하는 Bacillus amyloliquefaciens ATC6의 분리)

  • Lee, Se-Hyung;Chae, Jong-Pyo;Kim, Min-Jeong;Kang, Dae-Kyung
    • Journal of Animal Science and Technology
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    • v.52 no.1
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    • pp.65-70
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    • 2010
  • A bacterium producing acidic cellulase was isolated from pig feces. The isolate, ATC6 strain, was found to be Gram-positive, non-motile, catalase-positive, and spore-forming stain. Under an electron microscope, the cells were observed to be rod-shaped. The isolate was identified as Bacillus amyloliquefaciens ATC6 on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. Optimum pH and temperature for the cellulase activity of the culture supernatant of B. amyloliquefaciens ATC6 were found to be pH 4.5 and $55^{\circ}C$, respectively. More than 80% of its maximum activity was maintained at pH 4.0. The cellulase activity was maintained at temperatures ranging from 35 to $55^{\circ}C$ after 2 h incubation at pH 4.5, whereas it's activity decreased rapidly at $65^{\circ}C$.

Studies on the Effects of the Capacitation Methods of Spermatozoas on in-vitro Fertilization and Cleavage Rate of Bovine Follicular Oocytes (수정능획득 처리법이 소 난포란의 체외수정 및 분할율에 미치는 영향에 관한 연구)

  • 김상근;한성욱;한방근
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.125-132
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    • 1991
  • The studies on the carried out to investigate the effects of capacitation method of spermatozoa on the in vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and FCS for 24~48hrs in an incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18hrs with motile capacitated sperm by preincubation of mKRB, treatment of HIS(high strength ion), Ca-IA(Inophore A), BFF(bovine follicular fluids) and heparin. The results obtained in these experiments were summarized as follows : 1. The in vitro fertilizatin and cleavage rate offollicular oocytes fertilized with capacitated spermatozoas in BO solution by preincubation of mKRB, treatment of HIS, Ca-IA, BFF and heparin method were 53.1%, 33.9%, 50.8%, 48.1%, 58.8% and 28.1%, 17.7%, 26.2%, 22.8%, 32.8%, respectively. And the fertilization and cleavage rate of heparin method was of highest of all. 2. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solutin by both caffeine, BSA and heparin methods were 65.8%, 70.3% and 40.8%, 47.3%, respectively, and those rates were higher treatment of heparin+BSA, heparin+caffeine than treatment of heparin. 3. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoa in BO solution with heparin concentrations of 2, 5, 10, 20, 40$\mu\textrm{g}$/ml were 50.0%, 54.7%, 58.1%, 51.7% and 27.9%, 32.8%, 37.1%, 30.0%, respectively. And the fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution with 10$\mu\textrm{g}$/ml of heparin was the highest of all. 4. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with caffeine concentraton of 10, 20, 30, 40$\mu\textrm{g}$/ml were 71.4%, 74.3% and 70.6%, 70.0% and 45.7%, 47.3%, 44.1%, 41.4%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with caffeine and heparin together(70.3~74.3%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%). 5. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with BSA concentration of 5, 10, 20, 30$\mu\textrm{g}$/ml were 63.6%, 62.9%, 66.7%, 60.3% and 44.1%, 43.5%, 48.5%, 42.7%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with BSA and heparin together(60.3~66.73%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%).

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Production of Porcine Embryos in Different Culture Medium

  • Lee, S. Y.;Park, Y. H.;Park, C. K.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.67-67
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    • 2003
  • Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

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Acinetobacter pullorum sp. nov., Isolated from Chicken Meat

  • Elnar, Arxel G.;Kim, Min-Gon;Lee, Ju-Eun;Han, Rae-Hee;Yoon, Sung-Hee;Lee, Gi-Yong;Yang, Soo-Jin;Kim, Geun-Bae
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.526-532
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    • 2020
  • A bacterial strain, designated B301T and isolated from raw chicken meat obtained from a local market in Korea, was characterized and identified using a polyphasic taxonomic approach. Cells were gram-negative, non-motile, obligate-aerobic coccobacilli that were catalase-positive and oxidase-negative. The optimum growth conditions were 30℃, pH 7.0, and 0% NaCl in tryptic soy broth. Colonies were round, convex, smooth, and cream-colored on tryptic soy agar. Strain B301T has a genome size of 3,102,684 bp, with 2,840 protein-coding genes and 102 RNA genes. The 16S rRNA gene analysis revealed that strain B301T belongs to the genus Acinetobacter and shares highest sequence similarity (97.12%) with A. celticus ANC 4603T and A. sichuanensis WCHAc060041T. The average nucleotide identity and digital DNA-DNA hybridization values for closely related species were below the cutoff values for species delineation (95-96% and 70%, respectively). The DNA G+C content of strain B301T was 37.0%. The major respiratory quinone was Q-9, and the cellular fatty acids were primarily summed feature 3 (C16:1 ω6c/C16:1 ω7c), C16:0, and C18:1 ω9c. The major polar lipids were phosphatidylethanolamine, diphosphatidyl-glycerol, phosphatidylglycerol, and phosphatidyl-serine. The antimicrobial resistance profile of strain B301T revealed the absence of antibiotic-resistance genes. Susceptibility to a wide range of antimicrobials, including imipenem, minocycline, ampicillin, and tetracycline, was also observed. The results of the phenotypic, chemotaxonomic, and phylogenetic analyses indicate that strain B301T represents a novel species of the genus Acinetobacter, for which the name Acinetobacter pullorum sp. nov. is proposed. The type strain is B301T (=KACC 21653T = JCM 33942T).

The Marine Environment and Dinoflagellates Cysts in the Southwestern Sea of Korea (한국남서해역의 해양환경과 와편모조류 시스트 분포 특성)

  • Park, Jong-Sick;Yoon, Yang-Ho;Noh, Il-Hyeon;Soh, Ho-Young;Shin, Hyeon-Ho
    • ALGAE
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    • v.23 no.2
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    • pp.135-140
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    • 2008
  • A field survey for dinoflagellate cysts was carried out from May 2000 to November 2002 for the Southwest Sea of Korea. A total dinoflagellate cysts identified were 33 species, which belonged to 17 genera, 31 species, and 2 unidentified species. A cysts density were 16-1,501 cysts-gdry$^{-1}$. The dominant species of dinoflagellate cysts in the Southwestern Sea of Korea were Spiniferites bulloideus and Scrippsiella trochoidea, which are autotrophic species. To investigate the environmental characteristics of the Southwestern Sea of Korea using the dinoflagellate cysts, a principal component analysis (PCA) was conducted using the data collected from a total of 51 stations. From the score distribution map by the PCA, the Southwestern Sea of Korea was largely divided into three regions according to the first primary component and the second primary component. In other words, Group 1 was the western sea area of Mokpo and Jindo, Group 2 was the outer sea area of the South Sea, and Group 3 was the coastal areas of the South Sea around the Archipelago. It was found that this division of sea area was influenced by effects of the sea environment of the coastal areas of Korea. The coastal areas of Mokpo and Jindo that belong to Group 1 were affected by the cold Yellow Sea water. The outer sea area of the central parts of the South Sea that belong to Group 2, which is the boundary between the Southern coastal water of Korea and the Tsushima warm water, was subject to the formation of temperature fronts throughout the year, while Group 3 was affected by the coastal waters of Korea. It was also found that this division was in close relationship with the distribution of sediment facies in the bottom layer. From the above results, the environmental factors that influence the cyst distribution in he Southwestern Sea of Korea were found to include the eutrophication status of the sea area, the physical characteristics of the sea environment such as the flow of sea current and fronts, the sediment facies in the bottom layer, and the appearance volume of motile cells.

Studies on Preservation of Germ Cells in Hanwoo I. Effects of Frozen Thawed Viability and Characteristics in Bovine Semen (한우의 생식세포 보존에 관한 연구 I. 한우 정액의 일반성상 및 동결후 생존성에 미치는 영향)

  • 이명식;박정준;전기준;정영훈;우제석;박수봉;임석기;연성흠;손동수
    • Journal of Embryo Transfer
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    • v.18 no.2
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    • pp.143-149
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    • 2003
  • This study was carried out to cryopreserve and to investigate characteristics of semen in Hanwoo. Semen was obtained from bulls selected by Daekwanryeong Branch station. Semen was collected each morning of the experiment, placed in water jacketed tubes at 37$^{\circ}C$, and trans-ported to the research laboratory within 10 minutes. Semen was extended with Egg yolk-glycerol extender to contain 50${\times}$10$^{6}$ sperm/ml. Semen was cooled over a 6h period in water jacketed tubes from about 25 to 5$^{\circ}C$, Egg yolk-glycerol extender was added in one step at 5$^{\circ}C$. Semen was aspirated into 0.5ml straws, which were sealed with powder. Egg yolk-glycerol extender, which is used in Hanwoo sperm frozen and stored, semen from 13 Hanwoo bulls collected, the postthawed percentages of motile sperm were 65.7%. In semen characteristics of Hanwoo bulls, number of bulls volume are 5.7 ml and total cell count are 975${\times}$10$^{6}$ m1 ejaculate.

Isolation and Characteristics of Acetic Acid Bacteria for Persimmon Vinegar Fermentation (감식초 발효용 초산균의 분리 및 특성)

  • Park, Mi-Hwa;Lee, Jeong-Ok;Lee, Ju-Young;Yu, Seon-Joo;Ko, Yu-Jin;Kim, Yeong-Hoon;Ryu, Chung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1251-1257
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    • 2005
  • This study was carried out to investigate characteristics of acetic acid bacteria for persimmon vinegar fermen-tation. Acetic acid bacteria were isolated from statically fermented vinegar. Microscopically the cells appeared as non-motile and non-flagellated, preferentially occuring in pairs. Isolated strains were able to grow in the presence of acetic acid, ethanol, and glucose. Four of them produced 2-ketogluconic acid but did not produce 5-ketogluconic acid. The four strains isolated from statically fermented vinegar were considered to be grouped into Acetobacter. The strains were inoculated into persimmon juice for persimmon vinegar fermentation. They produced acetic acid in the range of 5.25$\∼$5.68$\%$.

Effects of Breeds, Insemination Time, Breeding Season, Sperm Concentration on Reproductive Performance of Sows Inseminated by Liquid Boar Semen (액상정액을 이용한 인공수정시 품종, 계절, 인공수정 횟수 및 정자농도가 번식성적에 미치는 영향)

  • Kim, In-Cheul;Park, Chang-Sik;Lee, Kyu-Seung;Seo, Kil-Woong;Han, Sung-Wook
    • Korean Journal of Agricultural Science
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    • v.28 no.2
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    • pp.85-91
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    • 2001
  • This study was carried out to investigate the effects of liquid boar semen on reproductive performance in swine artificial insemination. Many factors, which were breeds, time of insemination, breeding season, sperm per dose etc, have been tried to improve reproductive efficiency. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. This experiment was carried out from 1995 to 2000. There were no differences in the fertility results compared with 3 breeds (Landrace, Yorkshire and Duroc), frequencies of artificial insemination (double and triple) per estrus cycle and different seasons by using liquid boar semen. There were no significant differences in conception rate, farrowing rate and litter size using 4 trials of 3.0, 2.5, 2.0 and $1.5{\times}10^9/80ml$ in liquid boar semen with 70% of motile sperm cells. We confirmed that the sperm number per dose of $1.5{\times}10^9/80ml$ could be used for commercial artificial insemination.

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Summary on the Dinoflagellate Cyst Assemblages of Modern Sediments from Korean Coastal Waters and Adjoining Sea (한국연안해역 와편모조류 시스트 연구에 관한 고찰)

  • Yoon, Yang Ho;Shin, Hyeon Ho
    • Korean Journal of Environmental Biology
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    • v.31 no.4
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    • pp.243-274
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    • 2013
  • To investigate the research state and characteristics of dinoflagellate cyst along the Korean coastal waters, this work analyzed 45 studies conducted in the Korean coasts and basin, the south of Jeju, East China Sea including some part of the Yellow Sea. It was found that the research on dinoflagellate cyst began in the later half of the 1980s in Korea, and that the research areas focused on eutrophied regions where a red tide occurred or on the seas where coastal development occurred. In other words, no research on the East Sea was found. In terms of research contents, there were various studies on analysis of cyst assemblage, tracking of the changes in marine environment like process of eutrophication, roles as a seed population in occurrence of a red tide, creation of cyst by use of sediment trip and tracking of its changes, morphological changes by the change of pH concentration in the hypoxia zone of eutrophied region, germination of a specific species, and other international-level studies. Species composition and cell density also varied in the Korean coastal waters. However, much research on dinoflagellate cyst in Korea is not systematic, and is fragmentary and individual and hencesuffers from great limitations. The research results so far have focused only on specific regions, and sporadic research had been conducted by specific research groups. Dinoflagellate cyst of surface sediments represents the accumulation index of pelagic environment in the sea and is a useful index to understand the marine environment efficiently. In conclusion, Korea requires organized human resources and collaborative research on dinoflagellate cyst which in turn should be considered as a component of marine ecosystem and as an essential aspect of marine biology.

Studies on the Effects of the Follicle Size, Hormone Supplementation, Semen Type and Capacitation Method on In Vitro Maturation and Fertilization Rate of Bovine Follicular Oocytes (난포의 크기, 호르몬의 첨가, 정액의 형태 및 수정능획득 방법 등이 소 난포란의 체외성숙 및 체외수정율에 미치는 영향에 관한 연구)

  • 김상근;이만휘;이봉구;박항균
    • Korean Journal of Animal Reproduction
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    • v.14 no.4
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    • pp.237-244
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    • 1990
  • These studies were carried out to investigate the effects of the follicles size, hormone supplementation, semen types and capacitation methods on in vitro maturation and fertilization of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean Native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, ECS, BFF and MCC for 24~48hrs. in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 18~20hrs. with motile capacitated sperm in the TCF(Tyroide calcium-free) solution containing 200$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The oocytes classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 61.4%, 12.1%, 19.2%, 4.2% and 3.0% of the total oocytes recovered, respectively. The maturation and fertilization rate of the A, B, C class follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS were 89.1%, 78.0%, 52.6% and 78.1%, 66.1%, 33.3%, respectively. 2. The average number of the follicular oocytes recovered from follicles size, 1~2mm, 3~5mm and above 5mm in dimeter were 67, 98 and 63, respectively. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium were 56.7%, 82.5%, 46.0% and 44.8%, 71.4%, 28.6%, respectively. 3. The maturation and fertilization rate of follicular oocytes, cultured in the TCM-199 medium supplemented with 5%, 10%, 15%, 20% FCS and hCG, HCG, $\beta$-estradiol were 76.0%~82.3% and 26.2%~70.0%, and those values were higher the supplementation of the hormone than the non-supplementation. 4. The fertilization and cleavage rate of the follicular oocytes, inseminated with spermatozoas of epididymis cauda, neat and frozen semen were 63.3%, 73.3%, 70.0% and 32.7%, 37.8%, 38.3, respectively. 5. The fertilization and cleavage rate of follicular oocytes, fertilized with capacitated spermatozoas by heparin, BFF and HIS methods were 70.0%, 53.8%, 34.2% and 38.3%, 23.1%, 17.1%, respectively. And the fertilization and cleavage rate were higher method of heparin than other methods.r methods.

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