• Title/Summary/Keyword: streptomycin-resistance

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Expression of Clostridium thermocellum Endoglucanase Gene in Lactobacillus bulgaricus and Lactobacillus plantarum and in vitro Survival Characteristics of the Transformed Lactobacilli (Lactobacillus bulgaricus와 Lactobacillus plantarum 균주에서 Clostridium thermocellum 유래 endoglucanase의 발현과 발현 유산균의 in vitro 생존 특성)

  • Cho, J.S.;Kang, S.H.;Lee, H.G.;Lee, H.J.;Woo, J.H.;Moon, Y.S.;Yang, C.J.;Choi, Y.J.
    • Journal of Animal Science and Technology
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    • v.45 no.4
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    • pp.659-666
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    • 2003
  • Endoglucanase A from Clostridium thermocellum which is resistant to pancreatic proteinase was selected out of numbers cellulases then were expressed in lactobacilli. Recombinant lactobacilli expression vector, pSD1, harboring the endoglucanase gene from C. thermocellum under the control of its own promoter, was constructed. Both L. bulgaricus and L. plantarum were electrotransformed with pSD1. The endoglucanase activities of 0.120 and 0.144 U/ml were found in culture media of L. bulgaricus and L. plantarum containing pSD1, respectively. In vitro survival characteristics of the transformed lactobacilli were tested. Both L. bulgaricus and L. plantarum showed a similar resistance to low pH 3. Moreover, L. plantarum was bile-salt resistant in the presence of 0.3 and 1% oxgall. L. bulgaricus and L. plantarum showed a rather homogenous resistant pattern against the tested antibiotics. Both of the strains were resistant to amikacin, gentamicin, streptomycin, kanamycin, and colistin.

Physiological Characteristics and GABA Production of Lactobacillus plantarum K74 isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum K74의 GABA 생산 및 생리적 특성)

  • Park, Sun-Young;Shim, Hye-Young;Kim, Kee-Sung;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.143-152
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    • 2013
  • Aminobutyric acid (GABA), a major inhibitory neurotransmitter in the central nervous system of animals, has several physiological effects including anti-hypertensive, diuretic, tranquilizing, and anti-stress properties, in humans. The purpose of this study was to investigate Lactobacillus plantarum K74, which was isolated from kimchi and selected as a strain with a high ability to produce GABA, to develop a new starter culture for fermented milk production. L. plantarum K74 produced $134.52{\mu}g/mL$ GABA in MRS broth containing 1% MSG, $212.27{\mu}g/mL$ GABA in MRS broth containing 2% MSG, and $234.63{\mu}g/mL$ GABA in MRS broth containing 3% MSG. The optimum growth temperature of L. plantarum K74 was $34^{\circ}C$, reaching a pH of 4.4 after 18 hours of growth. L. plantarum K74 was most sensitive to novobiocin out of 16 different antibiotics tested, and was most resistant to kanamycin and polymyxin B. L. plantarum K74 did not produce ${\beta}$-glucuronidase, a carcinogenic enzyme, and was comparatively tolerant to bile juice and low pH. Furthermore, it displayed resistance to Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus at rates of 54.9%, 46.3%, and 0.7%, respectively.

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Prevalent Characteristics of Fowl Typhoid and Antibiotic Susceptibility of Salmonella gallinarum (닭 티푸스의 발생상황과 Salmonella gallinarum의 항균제 감수성)

  • 박노찬;도재철;조광현;장성준;권헌일;박덕상
    • Korean Journal of Veterinary Service
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    • v.18 no.2
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    • pp.113-123
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    • 1995
  • The present study was conducted to investigate the prevalent characteristics of Fowl Typhoid and antibiotic susceptibility of Salmonella gallinarum isolated from 56 infective or dead chickens of 20 egg laying farms in Kyung Buk province during the period from August to December 1994. 1. Among 416, 000 chickens of 92 flocks in 20 egg laying farms, 17, 360 chickens of 31 flocks were died of Fowl Typhoid. 2. Salmonella gallinarum was isolated from 56 chickens in liver and spleen, and then blood of infective chickens was positive to Pullorum antigen. 3, In the survey of gross lesion of 56 chickens, 43 chickens(76.8%) were swelled at liver, 39(69.6%) were swelled at spleen, 12(21.4%) were changed with bronze, 3(5.4% ) were hemorrhagic in peritoneal cavity. 4. In transmission pattern, 4 farms were outbreaked the entrance of chicken house at first, but the others were outbreaked at various place. They were transmitted at right and left directions in flock. 5. 2 farms confirmed at the early stage of infection were eradicated by removing infective chickens and administrating antibiotics, but 18 farms at chronic stage were not. 6. The biochemical properties of 112 Salmonella gallinarum from chickens were generally identical to those of the referance, but H$_2$S was not productive, cellobiose was fermentive. 7 Minimum inhibitory concentration(MIC) of 20 isolates was performed by using 21 antibiotics, MICs of Amikacin(Ak), Gentamicin(Gm), Kanamycin(Km), and Tetracycline (Tc) were below 1.6 ug/ml, Ampicillin(Am), Furazolidone(Fu) and Neomycin(Nm) were below 3.1 ug/ml, Cephalothin(Ce), Cefazoline(Cf) and Chloramphenicol(Cm) were below 6.3 ug/ml, Nalidixic acid(Na), Polymyxin(Po) and Rifampicin(Rf) were below 12.5 ug/ml, Penicillin (Pm) was below 25 ug/ml, Colistin(Co) and Streptomycin(Sm) were below 50 ug/ml, Sulfamerazine(Sr) and Sulfamethazine (St) were below 200 ug/ml, Lincomycin(Lm) and Spiramycin(Sp) were below 400 ug/ml, Bacitracin(Ba) was below 800 ug/ml. 8. Among the 20 isolates, all(100%) of those were sensitive to Ak, Am, Ce, Cf, Cm, Fu, Gm, Km, Na, Nm, Po, Rf, Sr, St and Tc, but 6 isolates(30%) were resistent to Co, 20(100% ) to Ba, Lm, Pm, Sm, and Sp. The drug resistance patterns were simple which 6 strains were BaCoLmPmSmSp type, and 14 were BaLmPmSmSp type.

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Genetic characterization of Shigella flexneri isolated from the diarrheic patients in Seoul region (서울지역 설사환자로 부터 분리된 Shigella flexneri의 성상과 유전적 특성)

  • Seung, Hyun-Jung;Kim, Moo-Sang;Oh, Young-Hee;Choi, Byung-Hyun;Chae, Hee-Sun;Chu, Jiaqi;Jun, Moo-Hyung
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.337-345
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    • 2006
  • The shigellae are common etiological agents of bacillary dysentery in humans and primates. During four years from 2002 to 2005, 22 strains of Shigella spp. were isolated from the diarrheic patients in Seoul region. All of them were identified as S. flexneri by biochemical tests and serotyping. The prevalence of serotypes were variable by year, but the major serotypes were 2a and 3a. In an antimicrobial susceptibility test, all of the isolates were resistant to streptomycin and tetracycline, and susceptible to amikacin, kanamycin, cefoxitin, and gentamicin. All of the isolates showed the multi-resistant patterns over 3 drugs. By analysis of the plasmid profile the isolates were classified into 7 groups (P1~P7). Serotypes 2a and 2b were distributed to P1, P2, P3, and P4. Serotype 3a was differentiated to P5 and serotype 3b, to P6 and serotype 4a, to P7. PCR results showed that all isolates were positive for two virulence genes, ipaH and ial, but none of the strains had stx gene. The set1A and set1B genes were detected from 12 isolates (54.5%) that belonged to serotype 2a and 2b. The sen gene was detected from 19 isolates (86.4%). The 22 isolates showed 12 to 17 DNA fragments in the sizes ranging from 20.5 kb to 1135 kb, resulting in 13 patterns by the PFGE with Not I digestion. The PFGE patterns of the isolates showed the close relation with the serotypes, but no relations with year of isolation and antimicrobial resistance.

The Pathogenic Role of Enterobacteria(Salmonella, Shigella and E. coli spp) Isolated in Korea (한국에서 분리된 장내세균(Salmonella, Shigella, E. coli 균속)의 병원적 역할에 관한 연구)

  • Jung, Tae-Hwoa;Lee, Yun-Tai;Lee, Myung-Won;Lee, Bok-Kwon;Kim, Ki-Sang
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.73-95
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    • 1986
  • A total of 5,462 isolates suspicious of Salmonella, Shigella and E. coli which were isolated during 1983 to 1985 by 12 City Hygine Laboratories and General Hospital Laboratories were received and identified at the National Salmonella Center, Seoul, Korea. The result of identification of these strains were summarized as follows: 1. It was confirmed that the total organisms broke down into 2,014 strains of Salmonella 1,294 of which were S. typhi, 887 strains of Shigella and 2,561 strains of E. coli. 2. For seasonal distribution of enteric pathogens, July was the month with the highest out breaks of salmonellosis, May was the month of Shigellosis, and April was of the highest month it in the case of E. coli. 3. Salmonella typhi with the highest incidence of isolation was shown to belong to various phage types, especially with the strains detected in Seoul. M1 type was widely distributed all over the country, but the majority was E1 type in 1983. 4. For age distribution of patients, the 20-29 age group had the highest incidence of salmonellosis whileas the 1 to 9 age group had the highest incidence of Shigellosis. 5. For sexuly distribution of Salmonella and Shigella infections seemed to be relatively higher in the female than in the male. However, E. coli. had no relationship to both sex. 6. The antibiotic sensitivity patterns of S. typhi cultures showed a tendancy to be resistant to colistin, gentamycin, neomycin, tetracycline and streptomycin. 7. The isolates of S. paratyphi-A, S. typhimurium and S. enteritidis seemed to have a tendency of multiple drug resistance. 8. 93.9 percent of 1,568 E. coli strains showed negative reactions to the antisera of enteropathogenic E. coli and 15.6 percent of them produced a heat-labile enterotoxin, but positive reaction to the antisera was 6.1 percent and 11.6 percent of them producled the enterotoxin.

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Identification of bacterial agents causing mastitis in dairy cattle and observation of residual changes of sulfadimethoxine in serum and milk of the cattle after administration of sulfadimethoxine sodium (유방염 우에서 원인균분리 및 sulfadimethoxine sodium 투여 후 혈청 및 유즙내 잔류량 추이)

  • 조민희;도재철;송희종;정종식
    • Korean Journal of Veterinary Service
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    • v.24 no.1
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    • pp.31-41
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    • 2001
  • This study was carried out to identify causative agents from california mastitis test(CMT) positive mastitic milk, and to examine the antimicrobial susceptibility of 50 heads in Seongju and Chilgok area of Gyeongbuk province. Sulfadimethoxine sodium(SMS) was intramuscularly administered once to four mastitis Infected dairy cattle at the rate of 10mg/kg body weight. After injection of SMS, the depletion rate of serum and milk from the cattle were periodically measured for five days. The CMT positive number from 50 heads showed 46% and that of 200 quarters was 47(23.5%). From 39 quarters of 47 heads 39 different microorganisms were identified. These organisms were classified into 12 species : Staphylococcus aureus 8(20.5%), Sta hemolyticus 6(15.4%), Streptococcus bovis 4(10.3%), Sta hyicus 3(7.7%), Sta epidemidis. Sta xylosus, Sta sciuri 2(5.1%), Str agalactiae 2(5.1%), Escherichia coli(10.3%), three Enterobacter cloacae(7.7%), two Ent aerogenes(5.1%) and one Salmonella spp(2.6%). As the results of antibiotic susceptibility test, gentamicin(Gm, 11 species 27 strains, 69.5%), cephalotin(Cf, 9 species 24 strains, 61.5%), sulfamethoxazole(Stx, 8 species 14 strains, 43.6%), tetracycline(Tc, 8 species 14 strains, 35.9%), and streptomycin(Sm), lincomycin(Lm), cefoperazon(Cp) and penicillin(Pc) have a highly resistance(7.7% ~5.1%). However, carbenicillin (Cb), amikacin(Ah) have no susceptible for all drugs. The mean residual concentration of SMS in serum according to the time lapsed were showed 33.964 $\pm$ 4.435ppm at the 4 hours after intramuscularly injection(AII). It was significantly(p<.05) decreased to 6.596 $\pm$ 3.402, 0.217 $\pm$ 0.119 and 0.005 $\pm$ 0.004ppm at the 1st, 3rd and 5th day AII. The mean concentration of SMS in milk was significantly(p<.05) decreased from 0.920 $\pm$ 0.42ppm to 0.084 $\pm$ 0.016ppm between 8 hours and 1 day AII. As the results of this experiments, sulfadimethoxine was residued at the level of no less than 0.01ppm in milk on the 2nd day AII. Thus, this results would be able to be used the basic index for prevention of sulfonamides residue in milk after treatment of dairy mastitis.

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A Epizootiological Study of Salmonella Infection on Piggery : II. A Study on Drug Resistance and R Plasmids in Salmonella (양돈장(養豚場)에 있어서 Salmonella 감염증(感染症)의 역학적(疫學的)인 연구(硏究) : II. Salmonella 속균(屬菌)의 약제내성(藥劑耐性) 및 전달성(傳達性) R plasmid)

  • Choi, Won-pil;Lee, Hi-suk;Yeo, Sang-geon;Lee, Hun-jun;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.26 no.2
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    • pp.229-235
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    • 1986
  • 1984년 5월부터 1985년 5월까지 대구(大邱), 경북(慶北), 경남(慶南) 및 충남지역(忠南地域) 7개(個) 양돈장(養豚場)의 자돈(仔豚) 및 성돈(成豚)의 분변(糞便) 및 양돈장(養豚場)의 흙, 하수(下水), 사료(飼料), 추비(推肥), 쥐 등 7,440예(例)와 대구시(大邱市) 도축장(屠畜場)의 도축돈(屠畜豚) 장간막임파절(腸間膜淋巴節) 및 직장내용물(直腸內容物) 555예(例)로부터 분리(分離)한 319주(株)의 Salmonelia속균(屬菌)을 대상으로 항균제(抗菌劑)에 대한 내성(耐性) 및 전달성(傳達性) R plasmid의 분포상황(分布狀況)을 조사(詞査)하였던 바 그 결과(結果)는 다음과 같다. 1. 공시균(供試菌) 319주(株) 중(中) 250주(株)(78.4%)가 ampicillin(An), chloramphenicol(Cm), gentamicin(Gm), kanamycin(Km), nalidixic acid(Na), rifampicin(Rf), streptomycin(Sm), sulfadimethoxine(Su), 또는 tetracycline(Tc)에 내성(耐性)을 나타내었으며, 약제별(藥劑別)로는 Su(74.9%), Sm(53.0%) 및 Tc(28.5%)에 높은 내성(耐性)이 인정(認定)되었다. 2. 내성균(耐性菌) 250주(株)의 전달성(傳達性) R plasmid 보유율(保有率)은 51.2%(128주(株))였으며, 약제별(藥劑別)로는 Am경우 100%, Tc 92. 3% 및 Cm 75.0% 순으로 보유율(保有率)이 높았다. 3. 내성균(耐性菌) 250주(株)의 내성양상(耐性樣相)은 SmSu(91주(株)), Su(59주(株)) 및 TcSmSu(50주(株))내성형(耐性型)이 대부분이었고 R plasmid 전달후(傳達後)의 내성양상(耐性樣相)은 TcSmSu(40주(株)) 및 TcSu(28주(株)) 내성형(耐性型)이 많았다. 4. 양돈장별(養豚場別) 내성균(耐性菌) 출현빈도(出現頻度)는 48.0~93.6%로 다양(多樣)하였고, 전달성(傳達性) R plasmid 보유율(保有率)은 0~77.8%로 내성균(耐性菌) 출현빈도(出現頻度)와 일치(一致)되지 않았다. 5. 공시균(供試菌) 319주(株) 중(中) 각각(各各) 2주(株)는 Rf 및 Gm에 대해 내성(耐性)을 나타내었고, 내성균(耐性菌) 250주(株) 중(中) 73.2%(183주(株))가 다제내성(多劑耐性)이었으므로 Salmonella의 다제내성화(多劑耐性化) 경향(傾向)이 있었다. 6. 내성균(耐性菌) 250주(株) 중(中) R plasmid 전달후(傳達後) 5주(株)는 TcAmCmSmSu내성형(耐性型), 1주(株)는 TcAmKmSmSu내성형(耐性型)임이 확인되었다.

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Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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