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Antibiotic Resistant Microbial Contamination (Enterobacter cloacae) Derived from Egg Yolk and Frozen Semen Extender in Porcine In Vitro Fertilized Embryos

  • Kwak, Seong-Song;Jeong, Se-Heon;Jang, Seung-Hoon;Jeon, Yu-Byeol;Nam, Young-Hee;Biswas, Dibyendu;Lee, Wan-Kyu;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.267-272
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    • 2010
  • The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin) and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium, frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer's instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin, cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin, kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.

Characterization of Bacillus thuringiensis Seven Isolates from Soil (토양에서 분리한 Bacillus thuringiensis 7균주의 특성)

  • 이형환;주대걸;강승철;임헌길
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.377-383
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    • 1992
  • Seven strains of Bacillus thuringiensis were isolated from soil in Korea and characterized. The isolates were named HL-8, 10, 12, 13, 14, 15 and 16 which produced parasporal crystals and endospores in their cells. The biochemical characteristics of the seven isolates were only minor different in specific chracteristics to the known serotypes of Bacillus thuringiensis. The number of the plasmid DNA elements from the isolates were studied. The computerized molecular weights of the six plasmid elements in the HL-8 and HL-lO strains were from 3.01 to 15.1 Md, four plasmid elements in the HL-12 were from 5.4 to 21.9 Md, four plasmid elements in HL-13 were from 5.1 to 20 Md, three plasmid elements in HL-15 were from 3.4 to 11.3 Md and three plasmid elements in the HL-16 were from 2.4 to 20.1 Md. The seven isolates showed resistances to ampicillin, bacitracin, cephalothin, methicillin and penicillin G. The strains of HL-8, HL-lO, HL12, HL-14, HL-15 and HL-16 showed lethalities against Culex pipiens 3rd instar larvae. The HL8 and 14 strains showed 100% lethality to the larvae within 48 hours. HL-13 strain did not have toxicity against the larvae.

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Characterization of Microbial Pathogen Bacillus thuringiensis Isolates from Soil Against Mosquito and Silkworm Larvae (II) (토양에서 분리한 살충성 Bacillus thuringiensis의 모기와 누에 유충에 대한 독성효과 (II))

  • Lee, Hyung-Hoan;Yoo, Bo-Rim;Kim, Young-Joo;Won, Nam-Hi;Kim, Hak-Chun
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.17-21
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    • 1993
  • Eight strains of Bacillus thuringiensis were isolated from soil in Korea and characterized. The isolates were named HL-24, HL-25, HL-33, HL-34, HL-35, HL-38, HL-39, HL-40. Strains HL-24 and HL-25 produced irregular parasporal crystals, HL-33 and HL-35 produced bipyramidal crystals, and others were round form in their cells. The biochemical characteristics of the eight isolates were only minor different in specific characteristics to the known serotypes of Bacillus thuringiensis. The HL-25, HL-33 and HL-34 strains showed resistances to cephalothin, colistin and penicillin G, and HL-39 and HL-40 strains were resistant to penicilin G. The strains of HL-24, HL-25, HL-33 and HL-34 were toxic to Bombyx mori lavae and HL-24, HL-25, HL-38, HL-39 and HL-40 strains killed Culex pipiens 3rd instar larvae. The HL-24 and H25 strains showed lethal activity against two kinds of the larvae, however lethality against mosquito larvae was low.

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Isolation and Antimicrobial Susceptibility of Campylobacter jejuni from Diarrhea Patients (설사환자에서 Campylobacter jejuni의 분리 및 항생제 감수성 특성)

  • Park, Eun-Hee;Kim, Joung-A;Choi, Seung-Hwa;Bin, Jae-Hun;Cheigh, Hong-Sik;Suk, Dong-Hee;Lee, Su-Chul;Kim, Young-Hui
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.811-815
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    • 2007
  • ln this study we isolated 27 isolates of Campylobacter jejuni from stool samples of 882 diarrheal patients. The seasonal distribution of patients was highest at July (11.7%). All the isolates of C. jejuni hydrolyzing sodium hippurate were serotyped on basis of heat-stable antigens, and identified with the use of passive hemagglutination assay. A total of 59.3% among 27 C. jejuni isolates were identified into 6 different serotypes, which serotype HS2, HSl/44, and HS2l were dominant. Antibiotics resistant rates of C. jejuni isolates were shown to be 100%, 63.0%, 51.9%, 37.0%, 33.3%, 25.9% and 7.4% to cephalothin, trimethoprim- sulfamethoxazole, tetracycline, ciprofloxacin, ampicillin, gentamycin and clindamycin, respectively. All isolates were sensitive to the erythromycin and imipenem.

Studies on properties and pathogenicity of Erysipelothrix rhusiopathiae isolated from pigs in Korea (국내(國內) 돼지에서 분리(分離)한 Erysipelothrix rhusiopathiae의 성장(性狀) 및 병원성(病原性) 관한 연구)

  • Yeh, Jae-gil;Seok, Ho-bong
    • Korean Journal of Veterinary Research
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    • v.31 no.2
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    • pp.201-208
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    • 1991
  • In studies on Erysipelothrix rhusiopathiae infection of pigs, a series of investigations were conducted for isolation, biochemical properties, pathogenicity, serotyping and susceptibility to antimicrobial agents. The results could be summarized as follows: 1. A total of 14 isolates of E rhusiopathiae were made from an acute form of swine erysipelas and the incidence was high during the hot summer season. 2. The biochemical properties of the 14 isolates were identical to the reference strain of E rhusiopathiae. 3. Pathogenicity of the isolated strain(89368) were ascertained that $LD_{50}$ for mice was $1.8{\times}10 $ cfu and also typical urticarious lesions and acute septicemia for pigs were induced by intradermal and intravenous inoculation with isolates, respectively. 4. All isolates were serotyped by the agarose gel slide double-diffusion system and proved to be serotype 1. 5. In susceptibility test to antimicrobial agents, 14 isolates of E rhusiopathiae were highly sensitive to ampicillin, baytril, cephalothin, chloramphenicol, penicillin and tetracycline.

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Bacteriological Study of Erysipelothrix rhusiopathiae Isolated from the Pigs in Youngnam Area (영남지방 돼지에서 분리한 Erysipelothrix rhusiopathiae의 세균학)

  • 백영숙;이진술;김영은;김봉환
    • Korean Journal of Veterinary Service
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    • v.14 no.1
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    • pp.49-61
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    • 1991
  • The present study was conducted to investigate biochemical, serologic, and pathogenic characteristic of E. rhusiopathiae isolated from the cases of acute septicemic swine erysipelas in Youngnam provinces during the period from June 1988 to September 1990. The majority of biochemical and cultural properties of E. rhusiopathiae isolated from pigs affected with acute erysipelas were identical to those of the standard strain employed. All of the 45 isolates were serotype la. All isolates were highly susceptible to penicillin G, lincomycin, cephalothin, ampicillin, erythromycin (MIC : 0.025-0.78IU or ${\mu}g$ / ml ), and moderately susceptible to oleandomycin, oxytetracycline, chloramphenicol (MIC : 0.78-25${\mu}g$ / ml ). Kanamycin and sulfadimethoxine showed no activity against the isolates(MIC : >400${\mu}g$ / ml ). The MICs of dihydrostreptomycin presented two distribution peaks ; of 45 strains, 5(11.1%) were resistant to dihydrostreptomycin(MIC : 400${\mu}g$ / ml ). All of 5 selected isolates were pathogenic for mite and $LD_{50}$ was $3.7{\times}10^3$viable cells. Mice immunized subcutaneously with live vaccine did not die after challenge to virulent isolates of E. rhusiopathiae.

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Distribution and Antibiotic Resistance of Vibrio vulnificus isolated in Incheon Coastal Area (인천지역 해양환경에서 분리된 비브리오패혈증군의 분포 및 항생제 내성 양성)

  • Hwang, Kyoung-Wha;Oh, Bo-Young;Gong, Young-Woo;Lee, Jae-Mann;Go, Jong-Myoung;Kim, Yong-Hee
    • Journal of environmental and Sanitary engineering
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    • v.22 no.4
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    • pp.55-63
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    • 2007
  • We investigated the distribution and drug susceptibility test of V. vulnificus isolated from environmental sources in Incheon. In this survey, total 4,302 samples were obtained from different sites of the Incheon coastal area during the periods from march 2004 to November 2006. Among the 4,302 samples, 310 strains were isolated. The isolation rates of Vibrio vulnificus from fish, shellfish, estuarine water and sediment were 6.7%, 4.7%, 12.4% and 23.2%, respectively. The highest isolation rate was 14.6% in September and in spite of low temperature the isolation rate was 5.1% in November. The highest rate of antibiotic resistance was observed against streptomycin(15.3%), cefazolin(8.5%), cephalothin(8.3%), amikacin(8.3%), cefoxitin(6.7%) and nalidixic acid(6.7%). Seventeen percent of isolates were observed to be resistant to two or more of the antibiotics tested.

Identification and Antimicrobial Susceptibility of Microorganisms Isolated from Bovine Mastitic Milk (젖소의 유방염 원인균 분리 및 약제 감수성 검사)

  • Kang, Hee-Jung;Kim, Ik-Chun;Kim, Jin-Hoe;Son, Won-Geun;Lee, Du-Sik
    • Korean Journal of Veterinary Research
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    • v.41 no.4
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    • pp.511-521
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    • 2001
  • Microorganisms were isolated and identified from bovine 296 quarters which showed positive reaction by California Mastitis Test (CMT) in 40 farms of Jeju from September 1999 to June 2000. The organisms associated with the mastitis of bovine were 11 different bacterial species in this study. Which of them, Staphylococcus aureus was the most predominant species as 152 (51.4%) isolates. Other identified species included 49 (16.5%) coliform, 47 (15.8%) Streptococcus dysgalactiae, 15 (5.1%) Bacillus spp., 8 (2.7%) Staphylococcus epidermidis, 6 (2.1%) Streptococcus agalactiae, 5 (1.7%) Enterococcus faecalis, 5 (1.7%) Corynebacterium spp., 3 (1.0%) Streptococcus uberis, 1 (0.3%) Pseudomonas aeruginosa and 1 (0.3%) Pasteurella haemolytica. Almost of all the islolated beacterial species showed high sensitivity against kanamycin (98.6%), cephalothin (98.0%), streptomycin (94.9%), gentamicin (94.6%), ampicillin (92.2%) and polymyxin B (90.2%). On the contrary, they showed resistance against penicillin (47.0%), tetracycline (37.2%), cefazolin (26.0%), bacitracin (22.6%) and erythromycin (19.9%). Eighty-one isolates were not resistant to any antibiotics and 215 drug resistant isolates showed 89 different drug resistance patterns from single to nine multiple antibiotics resistance patterns.

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Isolation of vibrio species from cultured flounders (paralichthys olivaceus) with uicers and ascites in the southern coast of Korea during the winter season (동절기 한국 남해안의 궤양증 및 복수증 양식 넙치로부터 vibrio종의 분리)

  • Lee, Hun-Ku;Kim, Hee-Je;Kim, Il
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.319-328
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    • 1991
  • this study was performed for the purpose of surveying the distribution and characteristics of the Vibrio species in the diseased farm flounders (Paralichthys olivaceus) during the winter season in Korea. Samples were collected along the southern coast in the Sinam, Keoje Island, and Teosu areas from November '90 to Feburary '91. Nine species of Vibrio were identified as V. tubiashii (85 strains), V. damsela (78 strains), V. anguillarum (21 strains), V. compbelli (9 strains), V. fluvialis (9 strain), V. costicola (1 strain), V. alginolyticus (1 strain), V. gazogenes (1 strain), V. marinus (1 strain), and unidentified 3 groups of bacteria (154 strains) which may have related with V. tubiashii were isolated. Eleven kinds of discs (BBL Co.) were used for antibiotic susceptibility test and 116 strains were selected for the experiment. Most bacteria were powerfully inhibited their growth by 4 antibiotics such as cephalothin, chloramphenicol, colistin, and tetracycline, but they were poorly inhibited by 4 antibiotics such as carbenicillin, kanamycin, penicillin G, and streptomycin.

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A Case Report of Proventriculotomy for Impaction of Proventriculus in Ostrich (타조에서 전위절개술을 통한 매복의 치유예)

  • 김남수;이종일
    • Journal of Veterinary Clinics
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    • v.16 no.2
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    • pp.469-473
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    • 1999
  • A 120 kg, 3-year-old, male ostrich (Struthio camelus) was submitted to the veterinary teaching hospital, Chonbuk national university, for chronic weight loss, anorexia, small hard fecal pellets, and lethargy. Illness was first noticed approximately 2 weeks before submission. Although the bird was able to stand, he was extremely weak and cachectic. Blood examination indicated that the bird was dehydration (PCV: 55%). Fluoroscopic and lateral abdominal radio-graphic view indicated impaction of distended proventriculus with bigger rocks than normal. Anesthesia was induced with ketamine (10 mg/kg, IV)-xylazine (1 mg/kg, IV) combination and maintained with ketamine (10 mg/kg, IV) at intervals of 8 to 15 minutes during the entire procedure. After induction of general anesthesia, the bird was positioned in dorsal recumbency. The skin was incised approximately 15cm caudal to the caudal margin of the sternum and extended 20 to 25 cm caudally. After the proventriculotomy was peformed, the impacted material that weighed 3kg contained rocks in 4 to 6cm diameter was removed. The incision of proventriculus was closed in 2 layers. Cephalothin sodium (30 mg/kg, IV, q12hrs) and dexamethazone (2 mg/kg, IM, q24hrs) was administered before and after 5 days of the operation. The bird was eating well and defecating normal feces after 3 weeks of the surgery

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