• 대한임상검사과학회지
• /
• 제38권1호
• /
• pp.26-33
• /
• 2006

Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

• 대한수의학회지
• /
• 제37권4호
• /
• pp.779-784
• /
• 1997

A comprehensive study of 132 Escherichia coli isolates from 150 piglets with colibacillosis included detection of heat-labile enterotoxin, heat-stable enterotoxin, and identification of K88 (F4), K99 (F5), 987P (F6), and F41. Four pili were examined by haemagglutination and slide agglutination test. Heat-labile(LT) and heat-stable(ST) enterotoxin was determined by reverse passive latex agglutination and precipitation test, respectively. Among 132 E coli isolates, 26 had K88 (19.7%), 16 had K99 (12.1%), 3 had 987P (2.3%), and 2 had F41 (1.5%). Three had K88 and K99 (2.3%), 3 had K88 and 987P (2.3%), 2 had K99 and 987P (1.5%), 5 had K99 and F41 (3.8%), and 8 E coli strains had K88, K99 and F41 (6.1%) simultaneously. Among 132 E coli isolates, 5 produced LT only (3.8%), 55 produced heat-stable toxin ST only (41.7%), and 4 produced both LT and ST (3.0%). Three major pathotypes accounted for 27.9% of E coli isolates: $K99^+$ (8.3%), $K88^+ST^+$ (9%) and $K88^+$ (10.6%). Results of this study indicated that piliated enterotoxin-producing E coli was prevalent and was associated with diarrhea in preweaning piglets.

• Journal of Microbiology and Biotechnology
• /
• 제21권2호
• /
• pp.204-211
• /
• 2011

To yield high concentrations of protein expressed by genetically modified Escherichia coli, it is important that the bacterial strains are cultivated to high cell density in industrial bioprocesses. Since the expressed target protein is mostly accumulated inside the E. coli cells, the cellular product formation can be directly correlated to the bacterial biomass concentration. The typical way to determine this concentration is to sample offline. Such manual sampling, however, wastes time and is not efficient for acquiring direct feedback to control a fedbatch fermentation. An E. coli K12-derived strain was cultivated to high cell density in a pressurized stirred bioreactor on a pilot scale, by detecting biomass concentration online using a capacitance probe. This E. coli strain was grown in pure minimal medium using two carbon sources (glucose and glycerol). By applying exponential feeding profiles corresponding to a constant specific growth rate, the E. coli culture grew under carbon-limited conditions to minimize overflow metabolites. A high linearity was found between capacitance and biomass concentration, whereby up to 85 g/L dry cell weight was measured. To validate the viability of the culture, the oxygen transfer rate (OTR) was determined online, yielding maximum values of 0.69 mol/l/h and 0.98mol/l/h by using glucose and glycerol as carbon sources, respectively. Consequently, online monitoring of biomass using a capacitance probe provides direct and fast information about the viable E. coli biomass generated under aerobic fermentation conditions at elevated headspace pressures.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권2호
• /
• pp.278-282
• /
• 2004

Compost is used mainly as an organic fertilizer, but it is also used as bedding material for cattle. Dairy cattle have been identified as a main reservoir of pathogenic Escherichia coli O157:H7. Further, E. coli is regarded as an environmental pathogen that causes bovine clinical mastitis. Hence, its growth in compost spread or compost bedding should be avoided. Physical and chemical conditions, available nutrients and microflora in compost change greatly during the composting process. Since pathogen growth in compost seems to be related to these changes, we assessed the possibility of E. coli growth in compost samples collected at 0, 7, 13, 22, 41, 190 and 360 d. Cattle waste composts with and without added tofu residue were collected from static piles and immediately air-dried. Compost samples were inoculated with a pure culture of E. coli, the moisture content was adjusted to 50%, and the samples were incubated for 5 d at $30^{\circ}C$. The numbers of E. coli in compost before and after incubation were determined by direct plating on Chromocult coliform agar. Almost all compost samples supported E. coli growth. Samples collected during or immediately after the thermophilic phase (day 7) showed the highest growth. Growth in samples more than 13 d old were not significantly different from those of aged compost samples. The addition of tofu residue gave a higher growth than its absence in younger samples collected prior to 13 d. To minimize the risk of environmental mastitis, the use of compost in the initial stage of the process is better avoided.

• 환경위생공학
• /
• 제19권4호
• /
• pp.69-75
• /
• 2004

The inhibitory effect of the food processing agent on growth of Escherichia coli O157:H7, Salmonella Enteritidis, and Listeria monocytogenes was performed with hydrogen peroxide and lactic acid, and combination of hydrogen peroxide and lactic acid. The minimun inhibitory concentration (MIC) of hydrogen peroxide in E coli O157:H7 was 100 ppm at pH 5.0, 6.0, 6.5 and 7.0, while in Listeria monocytogenes 25 ppm at PH 5.5, 6.0 and 50 ppm at PH 6.5, 75ppm at pH 7.0. MIC of lactic acid in E coli O157:H7 was 2500 ppm at pH 5.0, 6.0, 6.5 and 7.0. MIC of lactic acid in S. Enteritidis was 1250 ppm at pH 5.0, 2500 ppm at pH 5.5, 6.0, 5.5 and 7.0, while in L monocytogenes 625 ppm at pH 5.5 and 125 ppm at pH 6.0, 6.5 and 7.0. MIC of combined hydrogen Peroxide and lactic acid in E. coli O157:H7, S. Enteritidis, and L. monocytogenes was 75 ppm of hydrogen peroxide with 2500 ppm of lactic acid at pH 6.5. The correlations between MICs of hydrogen peroxide and lactic acid in E. coli O157:H7, S. Enteritidis and L. monocytogene were obtained through the coefficient of $determination(R^2)$. $R^2$ value were 0.9994, 0.9935 and 0.9283, respectively. The inhibitory effect of hydrogen peroxide and lactic acid in E. coli O157:H7, S. Enteritidis and L. monocytogenes could be confirmed from the result of this experiment. Therefore, it was expected that the food process would increase or maintain by using lactic acid together with hydrogen peroxide.

• Journal of Microbiology and Biotechnology
• /
• 제23권6호
• /
• pp.826-832
• /
• 2013

We investigated the transgalactosylation reaction of chlorphenesin (CPN) using ${\beta}$-galactosidase (${\beta}$-gal)-containing Escherichia coli (E. coli) cells, in which galactose from lactose was transferred to CPN. The optimal CPN concentration for CPN galactoside (CPN-G) synthesis was observed at 40 mM under the conditions that lactose and ${\beta}$-gal (as E. coli cells) were 400 g/l and 4.8 U/ml, respectively, and the pH and temperature were 7.0 and $40^{\circ}C$, respectively. The time-course profile of CPN-G synthesis under these optimal conditions showed that CPN-G synthesis from 40 mM CPN reached a maximum of about 27 mM at 12 h. This value corresponded to an about 67% conversion of CPN to CPN-G, which was 4.47-5.36-fold higher than values in previous reports. In addition, we demonstrated by thin-layer chromatography to detect the sugar moiety that galactose was mainly transferred from lactose to CPN. Liquid chromatography-mass spectrometry revealed that CPN-G and CPN-GG (CPN galactoside, which accepted two galactose molecules) were definitively identified as the synthesized products using ${\beta}$-gal-containing E. coli cells. In particular, because we did not use purified ${\beta}$-gal, our ${\beta}$-gal-containing E. coli cells might be practical and cost-effective for enzymatically synthesizing CPN-G. It is expected that the use of ${\beta}$-gal-containing E. coli will be extended to galactose derivatization of other drugs to improve their functionality.

• 한국미생물·생명공학회지
• /
• 제15권6호
• /
• pp.396-401
• /
• 1987

$\beta$-Galactosidase 유전자 조작된 두 대장균 숙주-벡터 시스템의 단백질 생산가 배양특성을 비교하였다. Tac 프로모터를 갖는 Escherichia coli JM109/ pTBG10 균주가 pL 프로모터를 갖는 E. coli MH 3000(pRKcI857)/ pASl(lacz) 균주보다도 단백질 생산성에 있어서 훨씬 우수하였으며 초기 및 중간 대수 증식기에서의 발현 유도가 단백질 생산에 있어서 유리함을 관찰하였다. 또한 유전자 발현시기에 있어서는 산소요구량이 매우 높았으나 pH를 일정하게 조절함에 의해 산소요구량을 어느 정도 낮출 수 있었고 생산성도 향상됨을 관찰할 수 있었다. 고생산성 균주(E. coli JM109/pTBG10)의 경우 플라스미드 함유 균체의 플라스미드 상실 균체에 대한 비증식속도의 비 $\mu$ + /$\mu$- 가 저생산성 균주보다도 낮았으며 이것은 유전자발현에 따른 단백질 합성부담이 매우 큰 때문으로 판단되었다. 또 플라스미드 안정성을 30세대까지 추적하였는바 유전자 발현 조건하에서 두 균주 모두 25세대 후 10% 정도의 안정성을 보였으나 E. coli JM109/pTBG10 균주의 안정성이 상대적으로 우수하다고 판단되었다.

• Journal of Microbiology and Biotechnology
• /
• 제26권2호
• /
• pp.270-276
• /
• 2016

Extended-spectrum β-lactamases (ESBLs), particularly those of the CTX-M types, are the predominant resistance determinants of Escherichia coli that are rapidly spreading worldwide. To determine CTX-M types, E. coli isolates were collected from retail chickens (n = 390) and environmental samples from chicken farms (n = 32) and slaughterhouses (n = 67) in Korea. Fifteen strains harboring bla_CTX-M genes were isolated from 358 E. coli isolates. The most common CTX-M type was eight of CTX-M-15, followed by six of CTX-M-1 and one of CTX-M-14. The bla_CTX-M genes were identified in the isolates from retail chickens (n = 9), followed by feces, water pipes, floors, and walls. Conjugations confirmed the transferability of the plasmids carrying bla_CTX-M genes to the recipient E. coli J53 strain. Furthermore, eight addiction systems carried by the replicons in CTX-M types were confirmed. The dominant system was identified as ccdAB, vagCD, and pndAC in donor strains and transconjugants. The clonal relationship between the two strains carrying bla_CTX-M genes indicates that E. coli may transmit from the farm to retail chickens, suggesting a possible public health risk. Our findings demonstrate that the detection of CTX-M types in E. coli isolates is important for tracking ESBL production in animals, and suggest linkage of multiple addiction systems in plasmids bearing bla_CTX-M genes.

• 한국토양비료학회지
• /
• 제44권3호
• /
• pp.368-370
• /
• 2011

가축분뇨 액비인 SCB 액비를 논토양 조건의 포트에 시용하였을 때 자외선에 의해 분변성 대장균의 사멸효과가 높은 것으로 나타났다. 따라서 본 연구결과로 볼 때 가축 분뇨의 퇴 액비화시 유해 미생물 제거에 광의 이용 가능성을 확인할 수 있었다. 그러나, 여러 종류의 가축분뇨 퇴 액비를 직접 농경지에 시용하였을 때 외부 환경의 건전성 유지와 환경 위해성을 관리하기 위해서는 향후 농경지 시용에 대한 현장 실험이 보완되어야 할 것이다.

• Journal of Veterinary Science
• /
• 제23권1호
• /
• pp.9.1-9.11
• /
• 2022

Background: Escherichia coli, which causes subclinical or clinical mastitis in cattle, is responsible for transmitting antimicrobial resistance via human consumption of raw milk or raw milk products. Objectives: The objective of this study was to investigate the molecular characteristics of 183 E. coli from bulk tank milk of five different dairy factories in Korea. Methods: The molecular characteristics of E. coli such as serogroup, virulence, antimicrobial resistance, and integron genes were detected using polymerase chain reaction and antimicrobial susceptibility were tested using the disk diffusion test. Results: In the distribution of phylogenetic groups, group D was the most prevalent (59.6%) and followed by group B1 (25.1%). The most predominant serogroup was O173 (15.3%), and a total of 46 different serotypes were detected. The virulence gene found most often was fimH (73.2%), and stx1, fimH, incC, fyuA, and iutA genes were significantly higher in isolates of phylogenetic group B1 compared to phylogenetic groups A, B2, and D (p < 0.05). Among 64 E. coli isolates that showed resistance to at least one antimicrobial, the highest resistance rate was observed for tetracyclines (37.5%). All 18 integron-positive E. coli carried the integron class I (int1) gene, and three different gene cassette arrangements, dfrA12+aadA2 (2 isolates), aac(6')-Ib3+aac(6')-Ib-cr+aadA4 (2 isolates), and dfrA17+aadA5 (1 isolate) were detected. Conclusions: These data suggest that the E. coli from bulk tank milk can be an indicator for dissemination of antimicrobial resistance and virulence factors via cross-contamination.