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Prevalence of chloramphenicol-resistant gene in Escherichia coli from water sources in aquaculture farms and rivers of Kuching, Northwestern Borneo

  • Leong, Sui Sien;Lihan, Samuel;Toh, Seng Chiew
    • Fisheries and Aquatic Sciences
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    • v.25 no.4
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    • pp.202-213
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    • 2022
  • Antibiotic resistant Escherichia coli cases are increasing high especially in Southeast Asia. Illegal use of the antibiotic in the aquaculture farming may become the culprit of the outbreak and spread into environmental source. A study was conducted to: 1) detect the chloramphenicol (CAL)-resistant gene in E. coli isolated from three aquaculture farms and six rivers of northwestern Borneo and 2) investigate the correlation between cat gene with five common antibiotics used. Isolation of E. coli was done on Eosin methylene blue agar and characterized using indole, methyl red, Voges-Proskauer, citrate tests. E. coli isolates were subsequently tested for their susceptibility to five antibiotics commonly used in aqua-farming. The CAL-resistant E. coli were further analyzed for the presence of resistant genes (cat I, cat II, cat III, cat IV) using multiplex polymerase chain reaction. 42 bacterial colonies were isolated from a total of 80 individual water samples, 34 of which were identified as E. coli. Result showed 85.3% of the E. coli isolates were resistant to amoxicillin, 35.3% were resistant to tetracycline, 29.4% were resistant to CAL, 17.6% were resistant to nitrofurantoin and 8.8% were resistant to nalidixic acid. All of the 10 CAL resistant E. coli isolateswere detected with cat II genes; five isolates detected with cat IV genes; three isolates detected with cat III genes; and another two detected with cat I genes. Pearson correlation coefficient shows highly significant relationship between resistance pattern of CAL with amoxicillin; and CAL with tetracycline. Our findings provide the supplementary information of the CAL resistance gene distribution, thereby improving our understanding of the potential risk of antibiotic resistance underlying within this microbial ecosystem.

Prevalence of CTX-M-type Extended-Spectrum $\beta$-Lactamases Producing Escherichia coli and Klebsieilla pneumoniae Isolates in General Hospitals in 2005 (임상에서 분리된 CTX-M형 Extended-Spectrum $\beta$-Lactamases를 생산하는 Escherichia coli와 Klebsiella pneumoniae의 유행)

  • Kim, Yun-Tae;Kim, Tae-Un
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.342-351
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    • 2006
  • The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.

Multiplication of Escherichia coli DH5α::gfp on Strawberry Fruit Surface (딸기과실 표면에서 Eschercia coli DH5α::gfp 증식)

  • Yun, Hyejeong;Park, Kyeonghun;Ryu, Kyoung Yul;Yun, Jong-Chul;Kim, Byung Seok
    • Food Science and Preservation
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    • v.20 no.2
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    • pp.250-256
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    • 2013
  • To verify the multiplication of microorganisms on the surface of strawberries, the fate of E. coli $DH5{\alpha}::gfp$ at different temperatures, times and strawberry extract concentrations were measured. The population of E. coli $DH5{\alpha}::gfp$ rapidly increased by 7.36~7.78 log CFU/g at $25{\sim}30^{\circ}C$ for 24 hr and slowly increased by 6.49~8.49 log CFU/g at $10{\sim}20^{\circ}C$ for 48 hr. However, E. coli $DH5{\alpha}::gfp$ did not grow at $10{\sim}15^{\circ}C$ on the surface of the strawberries, regardless of the contact times with the bacterial suspension. E. coli $DH5{\alpha}::gfp$ reached 1.52~3.26 log CFU/g at $20^{\circ}C$ as the contact frequency increased from two to six times. The contact frequencies did not significantly differ. In the case of the six-time contact on the surface of the strawberry at 25 and $30^{\circ}C$, the E. coli $DH5{\alpha}::gfp$ increased by 5.17 and 5.01 log CFU/g. The effects of the strawberry extracts on the growth of E. coli $DH5{\alpha}::gfp$ showed that sterilization and non-sterilization do not affect the growth of microorganisms for 96 hr. In the minimal broth, the growth of E. coli $DH5{\alpha}::gfp$ increased by 1 log CFU/g for 96 hr. In less than 50 percent of the strawberry extracts, the growth rate of E. coli $DH5{\alpha}::gfp$ was higher than in the control and increased by 4 and 5 log CFU/g at 50 and 25 percent of strawberry extracts, respectively. Therefore, E. coli $DH5{\alpha}::gfp$ can multiply and survive on the surface of strawberries when it comes into contact with the fruit extract.

Virulence Profile and Antimicrobial Resistance of Escherichia coli from Flies Captured from Agricultural Environment (농업환경에 서식하는 파리에서 분리된 E. coli의 병원성 유전자 및 항생제 내성 조사)

  • Yun, Bohyun;Jang, Youn Jung;Kim, Yeon Rok;Kim, Hwang-Yong;Kim, Won-Il;Han, Sanghyun;Kim, Se-Ri;Ryu, Jae-Gee;Kim, Hyun Ju
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.147-153
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    • 2017
  • The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.

Optimization of Electro-UV-Ultrasonic Complex Process for E. coli Disinfection using Box-Behnken Experiment (Box-Behnken법을 이용한 E. coli 소독에서 전기-UV-초음파 복합 공정의 최적화)

  • Kim, Dong-Seog;Park, Young-Seek
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.3
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    • pp.149-156
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    • 2011
  • This experimental design and response surface methodology (RSM) have been applied to the investigation of the electro-UV-ultrasonic complex process for the disinfection of E. coli in the water. The disinfection reactions of electro-UV-ultrasonic process were mathematically described as a function of parameters power of electrolysis ($X_1$), UV ($X_2$), and ultrasonic process ($X_3$) being modeled by use of the Box-Behnken technique, which was used for fitting 2nd order response surface model. The application of RSM yielded the following regression equation, which is empirical relationship between the residual E. coli number (Ln CFU) in water and test variables in coded unit: residual E. coli number (Ln CFU) = 23.69 - 3.75 Electrolysis - 0.67 UV - 0.26 Ultrasonic - 0.16 Electrolysis UV + 0.05 Electrolysis Ultrasonic + 0.27 $Electrolysis^2$ + 0.14 $UV^2$ - 0.01 $Ultrasonic^2$). The model predictions agreed well with the experimentally observed result ($R^2$ = 0.983). Graphical 2D contour and 3D response surface plots were used to locate the optimum range. The estimated ridge of maximum response and optimal conditions for residual E. coli number (Ln CFU) using 'numerical optimization' of Design-Expert software were 1.47 Ln CFU/L and 6.94 W of electrolysis, 6.72 W of UV and 14.23 W of ultrasonic process. This study clearly showed that response surface methodology was one of the suitable methods to optimize the operating conditions and minimize the residual E. coli number of the complex disinfection.

Chaperon Effects of Campylobacter jejuni groEL Genes Products in Escherichia coli (Campylobacter jejuni의 groEL 유전자 산물의 대장균에서의 Chaperon효과)

  • Lim, Chae-Il;Kim, Chi-Kyung;Lee, Jae-Kil
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.47-52
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    • 1994
  • The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

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Antibacterial Activity of Garlic Extract against Escherichia coli (마늘즙액의 대장균 생육 저해 작용)

  • Kim, Youn-Soon;Park, Kyung-Suk;Kyung, Kyu-Hang;Shim, Sun-Taek;Kim, Hyun-Ku
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.730-735
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    • 1996
  • Antibacterial action of garlic extract against Escherichia coli was investigated. When the survival of E. cloi in tryptic soy broth (TSB) containing 50% garlic extract was compared with those of Lactobacillus plantarum, Leuconostoc mesenteroides and Staphylococcus aureus, E. coli was the most sensitive to garlic antibacterial action. When E. coli was inoculated into TSB with different concentrations of garlic extract, viable cell number decreased continuously during the test period even at 1% garlic extract. When E. coli was inoculated into pH-adjusted TSB containing 0.5% garlic extract, viable cell number of E. coli decreased continuously at initial pH of 5.2 and 6.2, while it decreased initially but increased to $8.0{\times}10^{7}\;CFU/ml at 48 hr at pH 7.2. With larger initial populations $(10^{6}\;CFU/ml), E. coli grew without apparent inhibition, while with smaller initial populations $(<10^{5}\;CFU/ml), viable cell number decreased initially but later increased. Thiol compounds like cysteine and glutathione, with free SH group (s), helped E. coli to grow or survive better in TSB with inhibitory level (5%) of garlic extract. The possibility of eliminating E. coli by using garlic extract from foods like kimchi of which garlic is one of regular ingredients is suggested.

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Relationship Between Drug Resistance and Citrate Utilizing Ability of Citrate-Utilizing Variants of Escherichia Coli from Cattle (Citrate이용대장균(利用大腸菌) 변이주(變異株)의 약제내성(藥劑耐性)과 Citrate이용성(利用性)과의 관계(關係))

  • Lee, Hun Jun;Kim, Kyo Joon;Choi, Won Pil
    • Korean Journal of Agricultural Science
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    • v.10 no.2
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    • pp.200-205
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    • 1983
  • Transfer experiment of drug resistance and citrate utilizing ability were performed to show the relationship between drug resistance and citrate utilizing ability of 76 Citrate-Utilizing variants of Escherichia coli(Cit $^+E$. coli) isolated from cattle. The results obtained were summarized as follows; 1. Of seventy-six Cit $^+E$. coli, 36(47.4%) strains transfered their drug resistance to the E. coli ML 1410 by mating. 2. Tetracycline(TC) resistance determinants and citrate utilizing ability were transmitted in the recipient strains selected for TC, but it can be seen the segregation of streptomycin (SM) resistance determinants and citrate utilizing character in 9 recipient strains selected for SM. 3. Of seventeen TC resistance Cit $^+E$. coli, 10 strains transfered citrate utilizing character, alone. 4. The transmission of the ability to utilize citrate on Simmons citrate agar at $37^{\circ}C$, was demonstrated 52(68.4%) out of the 76 Cit $^+E$. coli.

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Effect of Organic Acids Derived from Black Liquor on Growth of Selected Escherichia coli MG 1655 (흑액 유래 유기산의 Escherichia coli MG 1655 성장에 미치는 영향 탐색)

  • Moon, Joon-Kwan;Um, Byung-Hwan
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.6
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    • pp.758-767
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    • 2014
  • In this study, we selected an Escherichia coli strain (E. coli MG 1655) metabolizing arabinose derived from acid hydrolyzed black liquor as a carbon source and investigated effect of organic acids (acetic acid, formic acid, and lactic acid) presented in black liquor on growth of the E. coli MG 1655. We measured growth of E. coli MG 1655 under various concentration of each and combined three kinds of organic acids. The E. coli MG 1655 shows tolerance to acetic acid, lactic acid and formic acid at these concentrations ($1.0g/{\ell}$ acetic acid, $1.2g/{\ell}$ lactic acid and $0.8g/{\ell}$ formic acid, respectively), but displays some growth retardation over $1.5g/{\ell}$ acetic acid, lactic acid $2.0g/{\ell}$, and formic acid $1.2g/{\ell}$, respectively. In addition, formic acid was shown to be a critical factor affecting growth of the E. coli MG 1655 in the presence of three kinds of organic acids. These results indicate that the inhibitors should be removed at least $1.0g/{\ell}$ of acetic acid, $1.2g/{\ell}$ of lactic acid, $0.8g/{\ell}$ of formic acid for normal cell growth required for high yield fermentation. In addition, there is a need to construct recombinant strains that may be resistant to the same or higher organic acids concentration (> $1.2g/{\ell}$) in the growth.

Molecular Characteristics of Extended-Spectrum β-Lactamase Genes in Clinical Isolates of Escherichia coli and Klebsiella pneumoniae (임상검체에서 분리된 Escherichia coli와 Klebsiella pneumoniae의 Extended-Spectrum β-Lactamase 유전자형 및 분자유전학적 특성)

  • Chung, Kyung-Suk
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.1
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    • pp.26-33
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    • 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.

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