scholarly journals Genotypic Analysis of Enterococci Isolated from Fecal-Polluted Water from Different Sources by Pulsed-Field Gel Electrophoresis (PFGE) for Application to Microbial Source Tracking

2011 ◽  
Vol 26 (2) ◽  
pp. 181-183 ◽  
Author(s):  
Takashi Furukawa ◽  
Hironori Takahashi ◽  
Terutoyo Yoshida ◽  
Yoshihiro Suzuki
Keyword(s):  
Gel Electrophoresis ◽  
Microbial Source Tracking ◽  
Pulsed Field Gel Electrophoresis ◽  
Source Tracking ◽  
Polluted Water ◽  
Pulsed Field ◽  
Genotypic Analysis ◽  
Different Sources

Listeria monocytogenes From Farm to Fork in a Brazilian Pork Production Chain

2020 ◽  
Vol 83 (3) ◽  
pp. 485-490 ◽  
Author(s):  
DANILO A. L. SILVA ◽  
CLARISSE V. BOTELHO ◽  
BRUNA T. F. MARTINS ◽  
RAFAELA M. TAVARES ◽  
ANDERSON C. CAMARGO ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Pulsed Field Gel Electrophoresis ◽  
Control Measures ◽  
Production Chain ◽  
Pork Production ◽  
Pulsed Field ◽  
Genetic Profiles ◽  
Different Sources

ABSTRACT Listeria monocytogenes contamination was assessed in different steps of a pork production chain. Ten lots of pigs were sampled at termination barns, at slaughter (after bleeding, after buckling, after evisceration, and after final washing), at processing (knives, deboning tables, and employees' hands), and of end products (ribs, shoulder, ham, and sausage). All samples (n = 670) were subjected to L. monocytogenes detection, and the obtained isolates (n = 18, identified as Listeria spp.) were characterized by their biochemical characteristics, serogroups, virulence genes, pulsed-field gel electrophoresis profiles, antibiotic resistances (ampicillin, penicillin, gentamicin, and sulfamethoxazole-trimethoprim), and adhesion abilities. The results revealed the low occurrence of Listeria spp. in the evaluated pork production chain. However, four tested sausage samples (40%) were positive for Listeria spp., with L. monocytogenes identified in two (20%) of these samples. Ten isolates were identified as L. monocytogenes (eight from serogroup 1/2a or 3a and two from serogroup 4b, 4d, or 4e): all isolates were also positive for the virulence-related genes hlyA, iap, plcA, actA, inlA, inlB, inlC, and inlJ and susceptible to the tested antibiotics. One sausage sample was contaminated by both serogroups 1/2a or 3a and 4b, 4d, or 4e. Isolates from serogroup 1/2a or 3a obtained during visits 5 and 6 presented distinct genetic profiles by pulsed-field gel electrophoresis, indicating that contamination may come from different sources. The adhesion potential exhibited by Listeria spp. isolates (n = 18) ranged from weak (serogroup 4b, 4d, or 4e) to moderate (L. innocua and L. monocytogenes serogroup 1/2a or 3a). Despite the low occurrence of L. monocytogenes, pathogenic serogroups were detected in sausages, demanding control measures by the industry. HIGHLIGHTS

scholarly journals Molecular Typing by Pulsed-Field Gel Electrophoresis of Spanish Animal and Human Listeria monocytogenes Isolates

2001 ◽  
Vol 67 (12) ◽  
pp. 5840-5843 ◽  
Author(s):  
A. I. Vela ◽  
J. F. Fernandez-Garayzabal ◽  
J. A. Vazquez ◽  
M. V. Latre ◽  
M. M. Blanco ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Samples ◽  
Pulsed Field ◽  
Clinical Strains ◽  
Great Genetic Diversity ◽  
Different Sources ◽  
Maize Stalks

ABSTRACT A total of 153 strains of Listeria monocytogenesisolated from different sources (72 from sheep, 12 from cattle, 18 from feedstuffs, and 51 from humans) in Spain from 1989 to 2000 were characterized by pulsed-field gel electrophoresis. The strains ofL. monocytogenes displayed 55 pulsotypes. The 84 animal, 51 human, and 18 feedstuff strains displayed 31, 29, and 7 different pulsotypes, respectively, indicating a great genetic diversity among the Spanish L. monocytogenes isolates studied. L. monocytogenes isolates from clinical samples and feedstuffs consumed by the diseased animals were analyzed in 21 flocks. In most cases, clinical strains from different animals of the same flock had identical pulsotypes, confirming the existence of a listeriosis outbreak. L. monocytogenes strains with pulsotypes identical to those of clinical strains were isolated from silage, potatoes, and maize stalks. This is the first study wherein potatoes and maize stalks are epidemiologically linked with clinical listeriosis.

scholarly journals Multiple-Locus Variable-Number Tandem Repeat Analysis ofVibrio choleraein Comparison with Pulsed Field Gel Electrophoresis and Virulotyping

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Cindy Shuan Ju Teh ◽  
Kek Heng Chua ◽  
Kwai Lin Thong
Keyword(s):  
Tandem Repeat ◽  
Gel Electrophoresis ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Multiple Locus ◽  
Repeat Analysis ◽  
Geographical Regions ◽  
Different Sources

Molecular analysis of MalaysianVibrio choleraewas carried out using a multiple-locus variable-number tandem repeat analysis (MLVA) assay based on 7 loci ofV. cholerae. The discriminatory ability of the assay was compared with pulsed-field gel electrophoresis (PFGE) using 43 MalaysianV. choleraeisolated from various sources. In addition, the virulotypes of the strains were determined. Based on MLVA, 38 allelic profiles were obtained (F=0.63) while PFGE generated 35 pulsotypes (F=0.71). Simpson's index of diversity for different VNTR loci ranged from 0.59 to 0.92. The combined loci increased the discriminatory index to 0.99 which was comparable with PFGE (D=0.99). Most of the environmental non-O1/non-O139 strains harboredrtxA,rstR,toxR, andhlyA only, and the virulotype of this serogroup was significantly different (P<.01) from clinical/environmental O1 and environmental O139 strains. In conclusion, the MLVA assay developed in this study was a useful genotyping tool with comparable discriminatory power with PFGE. In addition, the combination of the two approaches can further distinguish the strains from different sources and geographical regions of isolation.

scholarly journals Epidemiological Study of a Food-Borne Outbreak of EnterotoxigenicEscherichia coli O25:NM by Pulsed-Field Gel Electrophoresis and Randomly Amplified Polymorphic DNA Analysis

1998 ◽  
Vol 36 (3) ◽  
pp. 652-656 ◽  
Author(s):  
Toshihiro Mitsuda ◽  
Tetsunori Muto ◽  
Mikiko Yamada ◽  
Nobuyoshi Kobayashi ◽  
Masanori Toba ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Dna Analysis ◽  
Pulsed Field Gel Electrophoresis ◽  
Causal Agent ◽  
Randomly Amplified Polymorphic Dna ◽  
Pulsed Field ◽  
Food Borne ◽  
Length Polymorphisms ◽  
Genotypic Analysis ◽  
Epidemiological Characteristics

This study investigated the applicability of molecular epidemiological techniques to the identification of the causal agent of an outbreak of diarrhea caused by ingestion of food contaminated with enterotoxigenic Escherichia coli (ETEC). The outbreak occurred at four elementary schools in July 1996 and affected more than 800 people. Illness was most strongly associated with eating tuna paste (relative risk, 1.79; 95% confidence interval = 1.16 to 2.79; P = 0.0001). To evaluate the epidemiological characteristics of the pathogen, the DNAs from numerous isolated ETEC strains were subjected to randomly amplified polymorphic DNA analysis, pulsed-field gel electrophoresis of nuclease S1-treated plasmid DNA, and analysis of genomic DNA restriction fragment length polymorphisms. All ETEC isolates were of the O25:NM (nonmotile) serotype, which carries a heat-stable enterotoxin Ib gene. Genotypic analysis demonstrated that the strains isolated from the patients at all four schools were identical. The isolates of ETEC O25:NM obtained from the tuna paste that had been served for lunch at these schools were genetically indistinguishable from those isolated from the patients. Results suggest that this outbreak was food borne. The molecular biology-based epidemiological techniques used in this study were useful in characterizing the causal agent in this food-borne epidemic.

scholarly journals Genotyping of ESBL Producing UropathogenicEscherichia coliin West of Iran

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Parviz Mohajeri ◽  
Gita Darfarin ◽  
Abbas Farahani
Keyword(s):  
Gel Electrophoresis ◽  
Bacterial Infections ◽  
Pulsed Field Gel Electrophoresis ◽  
Molecular Fingerprinting ◽  
Pulsed Field ◽  
Predominant Type ◽  
Single Clone ◽  
Clonal Distribution ◽  
Sources Of Infection ◽  
Different Sources

Background and Objective.Urinary tract infection (UTI) is one of the most common bacterial infections in the world. Molecular fingerprinting of UTI isolates such as pulsed-Field Gel Electrophoresis using for Clonal distribution and determine of predominant type. The aim of the study was to determine genotyping of ESBL producing UPECs.Material and Methods.200 UPEC isolates from outpatients with UTI were obtained. Antimicrobial susceptibility and interpretation were performed by disk diffusion. Virulence factors for UPECs were screened by using PCR. UPECs were analyzed by Pulsed-Field Gel Electrophoresis and images analyzed by Phoretix1DPro software.Results.A total of 200 isolates of UPECs, 24.5% (n=49) of isolates, were positive for ESBL production. Resistance ranged from 0% for amikacin and imipenem to over 93.9% for carbenicillin and ampicillin. Frequencies of haemagglutination, haemolysin, and hydrophobicity were 51%, 18.3%, and 14.28%, respectively. A total of 10 different genotypes were obtained, which include nine common clones and one single clone.Conclusion.We confirmed the prevalence of virulence phenotyping especially Haemagglutination among UPEC strains and that it can also contribute to virulence in these strains. Large diversity in genotypes was observed in the isolates that could be indicative of different sources of infection in community acquired.

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