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Typing of Extended-Spectrum ${\beta}-Lactamase$ of Escherichia coli and Klebsiella pneumoniae Isolated from Rivers in Busan, Korea (부산지역 하천에서 분리된 장내세균 Escherichia coli와 Klebsiella pneumoniae의 광범위 베타 락탐 분해효소 (Extended-Spectrum ${\beta}-Lactamase$)에 대한 유형별 분류)

  • Lee, Hun-Ku;Kim, Hye-Jin;Kim, Gun-Do
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.116-123
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    • 2007
  • The purpose of this study was typing the plasmid mediated extended spectrum ${\beta}-lactamases$ produced by enteric bacteria isolated from rivers in Pusan. Six strains of Eschericha coli and fifteen strains of Klebsiella pneumoniae transferred their plasmid mediated extended spectrum ${\beta}-lactamase$ genes to the recipient strain Eschericha coli J53 $Azid^{R}$. The plasmid mediated extended spectrum ${\beta}-lactamase$ genes were sequenced directly after PCR and the types were determined by the BCM Search Launcher and GenBank nucleotid database. Determined types of the plasmid mediated extended spectrum ${\beta}-lactamases$ were TEM-52 and SHV-12. TEM-52 was isolated from both Escherichia coli and Klebsiella pneumoniae. However SHV-12 was isolated from Klebsiella pneumoniae only. The results indicated that the plasmid mediated extended spectrum ${\beta}-lactamase$ producing bacteria spreded over the area of clinical to the nature in Korea.

Survival of Escherichia coli O157:H7 and Salmonella ser. typhimurium in Fermented Milk Products (발효유제품내에서 Escherichia coli O157:H7과 Salmonella ser. typhimurium의 생존)

  • 김현욱;안영태;임정현;강호진;장영호
    • Journal of Food Hygiene and Safety
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    • v.12 no.3
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    • pp.175-180
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    • 1997
  • Escherichia coli O157:H7 and Salmonella ser. typhimurium are pathogens involved in food poisoning in numerous countries. This study aimed to obtain knowleges on the survival of E. coli O157:H7 KSC 109 and S. ser. typhimurium ATCC 14028 in fermentedmilk products which were on sale in Suwon Yakult supplier. To the final concentration of 103~104 cfu/$m\ell$ of E. coli O157:H7 KSC 109 or S. wer. typhimurium ATCC 14028 in the fermented milks, Metchnikoff, Ace, Yakult, Mastoni and Super 100 were inoculated with these pathogens and then were stored at 4$^{\circ}C$ and viable cells of these pathogens were periodically counted. The results showed that the survival of two pathogens differed in the different types of fermented milks tested. Number of suriviving E. coli O157:H7 KSC 109 and S. ser. typimurium ATCC 14028 cells (initial inoculum, 103~104 cfu/$m\ell$) were decreased to 101, 102 cfu/$m\ell$ in Ace after 100 hours, and were decreased gradually to 101 cfu/$m\ell$ in Yakult after 250 hours. In the other fermented milks, viable cells of E. coli O157:H7 KSC 109 was not drastically decreased but those of S. ser. typhimurium ATCC 14028 was decreased gradually to 102 (Mastoni), and to 101 cfu/$m\ell$ (Super 100) after 250 hours. It appeared that S. ser. typhimurium ATCC 14028 was more susceptible than E. coli O157:H7 KSC 109 at low pH. Vibale cells of E. coli O157:H7 KSC 109 was not drastically decreased in most of fermented milks tested except Ace and Yakult, but in general, S. ser. typhimurium ATCC 14028 was drastically decreased in most of the fermented milks. The major ingibition factor against these pathogens in the fermented milks during storage at 4$^{\circ}C$ appeared to be the acidity and the metabolites produced by the starters bacteria used in fermented milk products.

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Evaluating Commercial Spray Applications of Lactic Acid, Hot Water, and Acidified Sodium Chlorite for the Reduction of Escherichia coli on Beef Carcasses

  • Kang, Dong-Hyun;Lee, Sun-Young
    • Food Quality and Culture
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    • v.2 no.1
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    • pp.55-60
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    • 2008
  • This study examined the effects of lactic acid spray, hot water spray, or their combined treatment, as well as the effects of acidified sodium chlorite (ASC), for the decontamination of Escherichia coli on beef carcass surfaces using a commercial intervention system. With this system, the effects of 2 or 4% lactic acid (v/v), hot water ($89{\pm}1^{\circ}C$), or their combined treatment, were examined in terms of reducing inoculated E. coli. ASC (266 ppm), which was adjusted to pH 2.5 using acetic acid or citric acid, was applied using a hand-held spray system. When the beef carcasses were treated with 2 or 4% lactic acid for 10.4 s, less than 1 log reductions of inoculated E. coli were observed. A hot water spray treatment for 9.8 s resulted in a 2.1 log reduction of inoculated E. coli. However, when the hot water was followed with either 2 or 4% lactic acid, no difference in E. coli reduction was found between the hot water alone or the combined treatment with lactic acid. When ASC was adjusted to pH 2.5 with acetic acid and citric acid, 3.8 and 4.1 log reductions of E. coli were observed, respectively. Overall, the lactic acid spray treatment was least effective, and the ASC treatment was most effective, for the E. coli decontamination of beef carcasses. Therefore, these data suggest that ASC would be a more effective intervention against E. coli than most of the methods currently being used. However, more research is required to evaluate the effects of ASC on other organisms, as well as to identify application methods that will not affect meat quality.

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Application of Probabilistic Model to Calculate Probabilities of Escherichia coli O157:H7 Growth on Polyethylene Cutting Board

  • Lee, Joo-Yeon;Suk, Hee-Jin;Lee, Hee-Young;Lee, Soo-Min;Yoon, Yo-Han
    • Food Science of Animal Resources
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    • v.32 no.1
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    • pp.62-67
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    • 2012
  • This study calculated kinetic parameters of Escherichia coli O157:H7 and developed a probabilistic model to estimate growth probabilities of E. coli O157:H7 on polyethylene cutting boards as a function of temperature and time. The surfaces of polyethylene coupons ($3{\times}5$ cm) were inoculated with E. coli O157:H7 NCCP11142 at 4 Log $CFU/cm^2$. The coupons were stored at 13 to $35^{\circ}C$ for 12 h, and cell counts of E. coli O157:H7 were enumerated on McConkey II with sorbitol agar every 2 h. Kinetic parameters (maximum specific growth rate, Log $CFU/cm^2/h$; lag phase duration, h; lower asymptote, Log $CFU/cm^2$; upper asymptote, Log $CFU/cm^2$) were calculated with the modified Gompertz model. Of 56 combinations (temperature${\times}$time), the combinations that had ${\geq}$0.5 Log $CFU/cm^2$ of bacterial growth were designated with the value of 1, and the combinations that had increases of <0.5 Log $CFU/cm^2$ were given the value 0. These growth response data were fitted to the logistic regression to develop the model predicting probabilities of E. coli O157:H7 growth. Specific growth rate and growth data showed that E. coli O157:H7 cells were grown at $28-35^{\circ}C$, but there were no obvious growth of the pathogen below $25^{\circ}C$. Moreover, the developed probabilistic model showed acceptable performance to calculate growth probability of E. coli O157:H7. Therefore, the results should be useful in determining upper limits of working temperature and time, inhibiting E. coli O157:H7 growth on polyethylene cutting board.

Free Living Amoeba-Bacteria Interactions: Analysis of Escherichia coli Interactions with Nonpathogenic or Pathogenic Free Living Amoeba

  • Jung, Suk-Yul
    • Biomedical Science Letters
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    • v.17 no.1
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    • pp.7-12
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    • 2011
  • Free-living amoebae ingest several kinds of bacteria. In other words, the bacteria can survive within free-living amoeba. To determine how Escherichia coli K1 isolate causing neonatal encephalitis and non-pathogenic K12 interact with free-living amoebae, e.g., Acanthamoeba castellanii (T1), A. astronyxis (T7), Naegleria fowleri, association, invasion and survival assays were performed. To understand pathogenicity of free-living amoebae, in vitro cytotoxicity assay were performed using murine macrophages. T1 destroyed macrophages about 64% but T7 did very few target cells. On the other hand, N. fowleri which needed other growth conditions rather than Acanthamoeba destroyed more than T1 as shown by lactate dehydrogenase (LDH) release assay. In association assays for E. coli binding to amoebae, the T7 exhibited significantly higher association with E. coli, compared with the T1 isolates (P<0.01). Interestingly, N. fowleri exhibited similar percentages of association as T1. Once E. coli bacteria attach or associate with free-living amoeba, they can penetrate into the amoebae. In invasion assays, the K1 (0.67%) within T1 was observed compared with K12 (0%). E. coli K1 and K12 exhibited high association with N. fowleri and bacterial CFU. To determine the fate of E. coli in long-term survival within free-living amoebae, intracellular survival assays were performed by incubating E. coli with free-living amoebae in PBS for 24 h. Intracellular E. coli K1 within T1 (2.5%) and T7 (1.8%) were recovered and grown, while K12 were not found. N. fowleri was not invaded and here it was not recovered.

Detection of Escherichia coli O157:H7 Using Combined Procedure of Immunomagnetic Separation and Test Strip Liposome Immunoassay

  • Kim, Myung-Hee;Oh, Se-Jong;Durst, Richard-A.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.509-516
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    • 2003
  • A model system for the immnunochemical detection of Escherichia coli O157:H7 using a combined immunomagnetic separation (IMS) and test-strip liposome immunoassay (LIA) procedure was developed. Immunomagnetic beads coated with anti-E. coli O157 IgG antibodies were used to separate the E. coli O157 (including the H7 serotype) from culture. Immunoliposomes, whose surface was conjugated to goat anti-E. coli O157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used as a detection label. The test strip, onto which antibodies to goat IgG were immobilized, was the immunosensor capturing immunoliposomes that did not bind to E. coli O157:H7 on the immunomagnetic bead-E. coli O157:H7 complexes. In experiments, pure cell culture suspensions of $10^5 E.$ coli O157:H7 organisms per ml produced a measurable signal inhibition, whereas a weak yet detectable signal inhibition occurred with $10^3CFU/ml$. The inhibition signals increased, when the incubation time for IMS was extended to 90 min and higher IgG-tag density (0.4mol%) was used on the liposomes. With 0.2 and 0.4mol% IgG-tagged liposomes, the IMS-LIA procedure showed more improved signal inhibitions than those of a direct (no IMS) LIA. The combined assay, which measures the instantaneous signal from immunoliposomes, can be completed within 90 min, making it significantly faster than conventional plating methods and enzyme-linked immunosorbent assay (ELISA). Accordingly, it is quite feasible to use the combined immunoassay format of IMS and dye-loaded immunoliposomes for the detection of E. coli O157:H7.

The Influence of Lycii fructus Extracts on the Growth and Physiology of Microorganism (구기자(Lycii fructus) 추출물이 미생물 생육에 미치는 영향)

  • 주인선;성창근;오만진;김찬조
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.625-631
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    • 1997
  • This study was peformed to investigate the effects of Lycii fructus extracts on the growth and physiology of S. cerevisae D7l, L. casei KCTC 3165, and E. coli DH5. When Lycii fructus was added into solid culture media, the growth of S. cerevisea D7l and L. casei KCTC 3165 was increased, whereas that of E. coli DH5 was somewhat decreased. The growth of S. cerevisiea D7l and L. casei KCTC 3165 were much promoted in the culture media including methanol extract of 1.0% and 1.5%, respectively. E. coli DH5 was changed its morphology not only in 10% Lycii fructus juice but also 1.0% methanol and chloroform extract of Lycii fructus. Its size in those growth condition was eight times longer than that of the normal E. coli DH5. It was elucidated that elongation phenomenan of E. coli DH5 was also appeared by adding 0.135% of Na$^{+}$, $K^{+}$, and the mixture of Na$^{+}$ and $K^{+}$.TEX> +/.

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Effects of dietary lysozyme supplementation on growth performance, nutrient digestibility, intestinal microbiota, and blood profiles of weanling pigs challenged with Escherichia coli

  • Park, Jae Hong;Sureshkumar, Shanmugam;Kim, In Ho
    • Journal of Animal Science and Technology
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    • v.63 no.3
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    • pp.501-509
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    • 2021
  • The aim of this was evaluate the efficacy of lysozyme on growth performance, nutrient digestibility, excreta microflora population, and blood profiles of weanling pigs under Escherichia coli (E. coli) challenge. A total of 30 piglets weaned at 25 days, 7.46 kg body weight, were assigned to three dietary treatments, composed of five replications, two piglets per replication, for 7 days. The dietary treatment groups were negative control (NC; without antibiotics and lysozyme), positive control (PC; NC + antibiotics), lysozyme (NC + 0.1% lysozyme). All piglets were challenged orally with 6 ml suspension, containing E. coli K88 (2 × 109 CFU/mL). Dietary supplementation with lysozyme and PC resulted in no significant differences in average daily gain and gain to feed efficiency. Weanling pigs fed with E. coli challenge with lysozyme and PC treatments had significantly enhanced nutrient retentions of dry matter and energy (p < 0.05); however, there was a tendency to increase nitrogen digestibility. Furthermore, dietary inclusion of lysozyme and antibiotics treatment groups had a beneficial effect on excreta, ileal, and cecal of the fecal microbial population as decreased E. coli (p < 0.05) counts, without effects on lactobacillus counts. A significant effect were observed on a white blood cells, epinephrine and cortisol concentrations were reduced in piglets fed diets containing E. coli challenge with lysozyme and antibiotics supplementation comparison with the NC group. Therefore, the present data indicate that lysozyme in diet could ameliorate the experimental stress response induced by E. coli in piglets by decreasing intestinal E. coli, white blood cells and stress hormones and improving nutrient digestibility.

An Epidemiologic Study on Hosts and Pathogens of Urinary Tract Infection in Urban Children of Korea (2012-2017)

  • Yoo, Yeong Myong;Park, Byeong Sub;Lee, Shin Young;Park, Kyu Jung;Jung, Hyun Joo;Pai, Ki Soo
    • Childhood Kidney Diseases
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    • v.23 no.1
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    • pp.29-35
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    • 2019
  • Purpose: We aimed to determine characteristics of host, causative organisms, and antibiotic susceptibility of bacteria in pediatric patients with UTI living in metropolitan area of Korea. Methods: Retrospective investigation was done for the causative organisms of UTI in 683 pediatric cases treated at Ajou University Hospital from 2012 to 2017. Patients were classified into Escherichia coli and non-E. coli group, where E. coli group was subdivided into ESBL(+) and ESBL(-) groups based on whether the bacteria could produce extended spectrum beta-lactamase (ESBL). Antibiotic susceptibility of the causative organism was also determined. Results: A total of 683 UTIs occurred in 550 patients, of which 463 (67.8%) were first-time infection and 87 (32.2%) were recurrent ones (2-7 recurrences, 2.52 average), and 64.9% were male and 35.1% were female. The most common causative organism was E. coli (77.2%) and ESBL(+) E. coli was found in 126 cases. The susceptibility of E. coli to 3rd or 4th generation cephalosporin was relatively higher than that to ampicillin or amoxicillin/clavulanic acid. ESBL(+) E. coli showed higher resistance rate to 3rd or 4th generation cephalosporin than ESBL(-) E. coli. Conclusion: New treatment guideline should be considered due to the incidence of ESBL(+) E. coli increased up to one quarter of UTI cases.

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.