• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.385-393
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• 2016

Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50℃, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.

• Korean Journal of Microbiology
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• v.40 no.1
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• pp.1-6
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• 2004

Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

• Animal Bioscience
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• v.35 no.1
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• pp.138-146
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• 2022

Objective: The objective of the current study was to investigate the influences of conventional (CO) and deep litter (DE) systems on antimicrobial resistance in fecal Escherichia coli (E. coli). Methods: A cross-sectional study was carried out to detect antimicrobial resistance to E. coli in swine fecal samples in CO and DE systems located in western and northeastern Thailand. Individual rectal swab samples were taken only from healthy pigs. A total of 215 individual and healthy pigs were randomly selected for isolation and antimicrobial susceptibility test of E. coli by the disc diffusion method. The test panel included amoxicillin (AMX), colistin, doxycycline (DOX), enrofloxacin, gentamicin (GEN), kanamycin, neomycin (NEO), and trimethoprim-sulfamethoxazole (SXT). Results: There were significant (p<0.05) lower resistance levels for GEN, NEO, and SXT in the DE farms compared to those in the CO farms. There was a lower number of antimicrobial resistance agents (p<0.001) in the DE farms compared to those in the CO farms. This result was consistent with those in western (p<0.01) and northeastern (p<0.01) Thailand. Overall, antibiograms of AMX-SXT and AMX-DOX-SXT were found in the CO (19.09% and 20.91%, respectively) and the DE (16.19% and 24.76%, respectively) farms. No antimicrobial resistance (5.71%) was found and AMX (13.33%) resistant pigs in the DE farms, whereas the pattern of AMX-GEN-SXT (6.36%) and AMX-DOX-GEN-SXT (11.82%) resistant pigs was found in the CO farms. Conclusion: The DE system for pig farming was superior to conventional pig farming by lowering the resistance level of fecal E. coli to GEN, NEO, and SXT, with decreasing the number of antimicrobial resistance agents and inducing a small proportion of pigs to be free from antimicrobial resistance.

• IMMUNE NETWORK
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• v.2 no.4
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• pp.233-241
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• 2002

Background: Monoclonal antibodies (mAb) of rodent origin are produced with ease by hybridoma fusion technique, and have been successfully used as therapeutic reagents for humans after humanization by genetic engineering. However, utilization of these antibodies for therapeutic purpose has been limited by the fact that they act as immunogens in human body causing undesired side effects. So far, there have been several attempts to produce human mAbs for effective in vivo diagnostic or therapeutic reagents including the use of humanized xenomouse that is generated by mating knockout mice which lost Ig heavy and light chain genes by homologous recombination and transgenic mice having both human Ig heavy and light gene loci in their genome. Methods: Genomic DNA fragments of mouse Ig heavy and light chain were obtained from a mouse brain ${\lambda}$ genomic library by PCR screening and cloned into a targeting vector with ultimate goal of generating Ig knockout mouse. Results: Through PCR screening of the genomic library, three heavy chain and three light chain Ig gene fragments were identified, and restriction map of one of the heavy chain gene fragments was determined. Then heavy chain Ig gene fragments were subcloned into a targeting vector. The resulting construct was introduced into embryonic stem cells. Antibiotic selection of transfected cells is under the progress. Conclusion: Generation of xenomouse is particularly important in medical biotechnology. However, this goal is not easily achieved due to the technical difficulties as well as huge financial expenses. Although we are in the early stage of a long-term project, our results, at least, partially contribute the successful generation of humanized xenomouse in Korea.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.137-146
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• 1997

About Clostridium perfringens causing clinically necrotic enteritis or isolated from the intestinal contens of healthy chicken, We examined the usefulness of a rapid identification method by gas-liquid chromatography as well as the types of toxins. For this study, there were used 169 chickens including 116 broilers, 27 layers and 26 breeders which collected from 9 healty flock and 21 diseased flock showing necrotic enteritis. Among them, Cl perfringens was isolated from 30 chickens(17.8%) including 7 breeders(26.9%), 5 layers(18.5%) and 18 broilers(15.5%). Isolation of Cl perfringens was mainly from ceca (100%) and followed by small intestines(70.0%) and livers(16.7%), respectively. Average concentration of the pathogen in intestinal contents was $10^{3.8}CFU/g$ in cases occuring necrotic enteritis and on the contrary $10^{3.8}CFU/g$ in healthy cases. All isolates tested showed the same characterstics in biochemical tests compared to those in standard strain. Analysis of gas-liquid chromatography to volatile fatty acids produced by Cl perfringens in PYG broth showed the typical peaks of acetic and butyric acids compatible with the standard chromogram and was confirmed as a effective and reliable tool for rapid identification of the bacteria. Toxin types of 30 strains were mostly classified in A type(26 isolates) and the rest in C type(2 isolates) and unidentifed type(2 isolates). All the isolates were highly susceptible to amphicillin, amoxicillin and cephalothin.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.5
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• pp.478-483
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• 2011

An antimicrobial material was purified from the acidified whole body extract of the jellyfish Nemopilema nomurai by using C18 reversed phase and cation-exchange high performance liquid chromatography (HPLC). Whole body extract and the purified compound (JAP-1) showed potent antimicrobial activities against a wide range of microorganisms including Escherichia coli D31, Bacillus subtilis, Streptococcus iniae and Candida albicans, without significant hemolytic activity. Treatment of JAP-1 with trypsin completely abolished all antibacterial activity against Bacillus subtilis, suggesting that JAP-1 is likely to be a proteinaceous antibiotic. The molecular weight of JAP-1 was determined to be 680.10 Da by MALDI-TOF mass spectroscopy.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.381-385
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• 2011

Methicillin-resistant Staphylococcus aureus (MRSA) is one of major pathogen causing hospital infection and several diseases such as purulent infection, bacteremia. The isolation ratio of MRSA is gradually increased up to 80% in the hospital, which makes a limitation for treatment of antibiotics because the isolated MRSA show resistance to methicillin as well as other antibiotics. This study proposes that mecA detecting methods which are not commonly used because of cost in the hospital is a more accurate method than Susceptibility Testing to detect a MRSA. We compared Staphylococcus aureus ATCC 29213 as a negative control and 20 MRSA strains isolated from patients by these two methods. We amplified mecA gene by polymerase chain reaction (PCR) and confirmed the PCR products by sequencing. All of the MRSA showed oxacillin and cefoxitin resistance whereas 85% (16/19) of the strains had mecA wildtype. These results suggest that some of the MRSA are mecA mutants therefore mecA genotyping reinforces the MRSA detection by antibiotic susceptibility test.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.255-260
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• 1994

원시생식세포(primordial germ cell; PGC)는 성성숙 이후에 기능을 갖는 생식세포의 근원이 되는 세포로서, 다능성을 갖고 있는 것으로 알려져 있다. 그러므로 chimera 및 유전자 변환동물 생산을 위해 널리 사용되어 온 배아주(embrynic stem; ES)세포를 대신할 다른 세포계라고 생각되어져 많은 연구가 진행되고 있다. 본 실험은 체외배양을 통하여 원시생식세포의 증식과 확립을 위해 배양조건을 구명하고, 또한 성장인자의 효과를 검증하기 위하여 실시되었다. 원시생식세포는 12.5일째의 ICR 생쥐태아의 원시생식선 융기조직으로부터 추출하였으며, DMEM + 20% FCS + nucleosides + antibiotics로 조성된 sDMEM 배양액을 사용하여 mitomycin C로 전처리한 되먹임세포단층(feeder layer)위에서 체외배양하였다. bFGF 및 LIF를 20, 40ng/ml농도로 각각 또는 함께 첨가하여 성장인자의 효과를 검토하였다. 원시생식세포는 성에 따라 유의적인 colony 형성율을 보였고(♂:1.9 colonies / genital ridge, ♀:1.3 colonies / genital ridge), bFGF 및 LIF의 첨가 및 첨가농도에 따라서도 유의성 있는 결과를 보였다(0.3~1.9 colonies / genital riege). 그러나 3회 이상 계대배양을 할 경우, 원시생식세포의 colony를 4% prarformaldehyde로 20분간 고정한 후, tris-maleate buffer(pH 9.0)로 10분간 3회 세정하였다. Fast Red로 염색을 실시한 결과, 대부분의 colony가 염색반응을 보여 다능성을 갖는 원시생식세포의 colony임이 입증되었다. 그러나 대부분의 colony가 3회 이상의 계대배양시 생종율이 급격히 떨어지는 것을 감안하면, 또 다른 미지의 성장인자나 보다 적절한 배양조건이 요구된다고 생각된다.

• Korean Journal of Veterinary Service
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• v.26 no.2
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• pp.121-128
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• 2003

This study was carried out to identify and investigate antimicrobial susceptibility for Mannheimuia haemolytical which is responsible for shipping fever. Samples were collected from nasal and lung of 100 adult healthy cattle which are slaughtered in Samsung meat corporation located in Incheon metropolitan city. lung lesion index have been investigated within 0-5 range according to Shewen and Willkie(Can J Vet Res 52:30-36, 1988). Eighty-seven of 100 cattle were under normal condition with 0-1 ranges. A total of 129 strains were collected from blood and tryptic soy agar. Among these strains, 100 strains were identified with Staphylococcus, Streptococcus and enterobacteria containing E coli. Biochemical and fermentation assay of arabinose, trehalose, xylose, mannose, mannitol, lactose and salicin were tested to identify with Mannheimia sp. for 7 strains shown haemolytic activity on blood agar. Five strains were identified with Mannheimia haemolytica and 2 strains were untyped. In seasonal survey, Mannheimia sp recovered from fall to winter(5 of 7) have been highly isolated rather than those from spring to summer(2 of 7). Mannheimiz haemolytica were susceptible to antibacterials tested in this study but more resistant to oxytetracycline and streptomycin.

• Korean Journal of Veterinary Service
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• v.21 no.4
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• pp.451-465
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• 1998

The present study was carried out to investigate the prevalence of brucellosis in Kyungbuk area for the 3 years from 1966 to 1998. Collective milk samples were routinely screened to detect positive farms by using the milk ring test(MRT), and serum agglutination test was performed to detect sero-positive individuals in the MRT positive farms. Attempt were made to isolate the causative organismas from slaughtered sero-positive reactors and some biochemical and polymerase chain reation characters of the isolates were also made to identify the organisms. Seroprevalence to brucellosis in peoples who are close contact with infected dairy herds was also investigated. Brucellosis of dairy cattle was rare before 1997, but has been broken more frequently since early 1998. By the MRT for dairy herds, positive rate was gradually increased every year : 0.6% in 1996, 1.5% in 1997, 3.9% in 1998. Among 262 MRT-positive herds, only 21 herds(8.0%) showed positive brucellosis in serological test. The isolation rates of Brucella sp from tested materials were 51.2% in supramammary glands, 39.5% in milks, and 50.0% in pulmonary Iymphnode, respectively. Isolated strain and biotype were Brucella(B) arbortus biotype 1 in 26 heads, and were B suis biotype 1 in 2 heads. Isolated strain and vaccine strain were very similar in their colony morphology and staining. In drug susceptibility, isolated stains(B abortus) and vaccine strain(B abortus RB-51) were sensitive to ampicillin, gentamycin, kanamycin, neomycin, penicillin, streptomycin, and to tetracycline, but resistant to erythromycin. In the PCR, field strains reacted to BA and IS711 primers, and vaccine strain reacted to BA, IS711, and RB5l primers. In the plate agglutination test of 96 sera of human contacted with animals, serum antibody titer detected 1 : 100 in one person, 1 : 200 in one, and below 1 : 25 in the others.