Molecular Subtyping and Source Attribution of Campylobacter Isolated from Food Animals

2016 ◽  
Vol 79 (11) ◽  
pp. 1891-1897 ◽  
Author(s):  
GREGORY H. TYSON ◽  
HEATHER P. TATE ◽  
JASON ABBOTT ◽  
THU-THUY TRAN ◽  
CLAUDINE KABERA ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Poultry Meat ◽  
Food Sources ◽  
Clinical Samples ◽  
Source Attribution ◽  
Control And Prevention ◽  
Retail Meat ◽  
Human Campylobacteriosis ◽  
Campylobacter Spp

ABSTRACT Campylobacter spp. commonly cause gastrointestinal illness in humans. Poultry meats have long been considered the predominant source of these infections, but few in-depth Campylobacter source attribution studies have been completed. We analyzed more than 1,300 Campylobacter isolates recovered from a number of animal and food sources, including dairy and beef cattle, pigs, poultry, and retail poultry meat, and compared them with Campylobacter isolates recovered from human clinical samples. Each isolate was subtyped using pulsed-field gel electrophoresis (PFGE) with SmaI and queried against the Centers for Disease Control and Prevention PulseNet database to identify human isolates with indistinguishable patterns. Half (49.5%) of the PFGE patterns from poultry animal and retail meat isolates were indistinguishable from patterns of at least one human isolate. Among the isolates from beef and dairy cows, 56.6 and 65.0%, respectively, of their PFGE patterns were indistinguishable from those of human isolates. Only a small portion of the PFGE patterns of Campylobacter isolated from pigs (9.5%) were found to have PFGE patterns in common with human isolates. These data imply that cattle may be larger contributors to Campylobacter infections than previously recognized and help further our understanding of potential sources of human campylobacteriosis.

Low Potential Virulence Associated with Mutations in the inlA and prfA Genes in Listeria monocytogenes Isolated from Raw Retail Poultry Meat

2013 ◽  
Vol 76 (1) ◽  
pp. 129-132 ◽  
Author(s):  
VICTORIA LÓPEZ ◽  
JAIME NAVAS ◽  
JOAQUÍN V. MARTÍNEZ-SUÁREZ
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Pulsed Field Gel Electrophoresis ◽  
Poultry Meat ◽  
Pathogenic Potential ◽  
Molecular Serotyping ◽  
Potential Source ◽  
Raw Foods ◽  
Over Time

Packaged raw foods can represent a potential source of Listeria monocytogenes contamination when opened at home, and listeriosis is associated with the consumption of undercooked raw foods. The aim of this study was to characterize a group of L. monocytogenes strains isolated from 56 packages of raw chicken meat from a single brand in order to determine the diversity of the strains that dominate in a particular food over time, as well as their pathogenic potential. Forty (71%) samples were found to be positive for L. monocytogenes, and three isolates per sample were subjected to PCR molecular serotyping. Subtyping of 45 isolates from different manufacturing dates (n = 40) or different molecular serotype within the same sample (n = 5) identified 11 different L. monocytogenes subtypes as defined by pulsed-field gel electrophoresis and sequencing of virulence genes actA and inlA. Two of the subtypes accounted for 51% of the isolates. About 40% of isolates (three subtypes) were found to potentially present attenuated virulence because of the presence of mutations in the prfA and inlA genes.

scholarly journals Heterogeneity of Salmonella isolates obtained from various sources in Russia 2010–2019

2020 ◽  
Vol 25 (1) ◽  
pp. 26-34
Author(s):  
Sofia Sh. Rozhnova ◽  
Konstantin V. Kuleshov ◽  
Anastasia S. Pavlova ◽  
Anna N. Guseva ◽  
Tatiana A. Kozhakhmetova ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Regional Differences ◽  
Significant Proportion ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Samples ◽  
Pork Meat ◽  
Salmonella Spp ◽  
The Russian Federation ◽  
Meat Samples ◽  
Transmission Factors

Aim: the goal of the study was to evaluate the heterogeneity of the Salmonella enterica subsp. enterica strains isolated from clinical specimens and various environmental sources in the Russian Federation during the period 20112019. Materials and methods. The data of 3076 non-typhoid isolates of Salmonella obtained from sporadic and outbreak cases of salmonellosis (n = 2518), food and water samples (n = 558) were used. These isolates were serotyped according to the KaufmanWhite scheme and genotyped by Pulsed-Field Gel Electrophoresis (PFGE) using XbaI and BlnI restriction endonucleases according to a standard PFGE-protocol developed by PulseNet International Network. Results. The studied Salmonella isolates were differentiated into 73 serotypes and 601 PFGE types. A comparative analysis of isolates from various sources made it possible to identify subtypes that differed significantly in their prevalence in humans and potential transmission factors (sources). A significant proportion of chicken, turkey, and pork meat samples contained PFGE-subtypes which did not occur in clinical samples. Regional differences in the heterogeneity of the Salmonella spp. were also identified. Conclusions. Genetic heterogeneity of the Salmonella population from humans and other sources shows significant variability of virulent properties and indicates the necessity of differentiated assessment of their epidemiological potential.

scholarly journals Detection of the Philadelphia chromosome in acute lymphoblastic leukemia by pulsed-field gel electrophoresis

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 1101-1107 ◽  
Author(s):  
AL Hooberman ◽  
CM Rubin ◽  
KP Barton ◽  
CA Westbrook
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Gel Electrophoresis ◽  
Chromosomal Abnormalities ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
Pulsed Field Gel Electrophoresis ◽  
Myelogenous Leukemia ◽  
Clinical Samples ◽  
Pulsed Field ◽  
Translocation Breakpoints

Abstract The Philadelphia (Ph1) chromosome is an acquired abnormality in the malignant cells of 10% to 25% of patients with acute lymphoblastic leukemia (ALL). Unlike chronic myelogenous leukemia (CML), where the molecular detection of the Ph1 chromosome is relatively straightforward using conventional Southern hybridization analysis, the detection of the Ph1 chromosome in ALL is complicated by the existence of several molecular subtypes, and the fact that translocation breakpoints are dispersed over a large genomic area. To circumvent these difficulties, we investigated pulsed-field gel electrophoresis (PFGE) to determine if this method could be used directly on clinical samples to detect the Ph1 chromosome in ALL. We report that, in a study of seven patients with Ph1-positive ALL, we could easily detect the Ph1 using only a single PFGE analysis, regardless of the Ph1 subtype, and we could confirm that the translocations occur either within or very near the BCR gene in all seven. We conclude that PFGE is a useful technique for the detection of the Ph1 in ALL, which ultimately may find wide applicability in the detection of other chromosomal abnormalities in other malignancies.

Evaluation of Logistic Processing To Reduce Cross-Contamination of Commercial Broiler Carcasses with Campylobacter spp.

2007 ◽  
Vol 70 (11) ◽  
pp. 2549-2554 ◽  
Author(s):  
LAKSHMI-PRASANNA POTTURI-VENKATA ◽  
STEFFEN BACKERT ◽  
SERGIO L. VIEIRA ◽  
OMAR A. OYARZABAL
Keyword(s):  
Gel Electrophoresis ◽  
Food Production ◽  
Pulsed Field Gel Electrophoresis ◽  
Cross Contamination ◽  
Large Problem ◽  
Pulsed Field ◽  
Before And After ◽  
Commercial Broiler ◽  
Simple Logistic ◽  
Campylobacter Spp

Cross-contamination of broiler carcasses with Campylobacter is a large problem in food production. Here, we investigated whether the contamination of broilers carcasses from Campylobacter-negative flocks can be avoided by logistic scheduling during processing. For this purpose, fecal samples were collected from several commercial broiler flocks and enumerated for Campylobacter spp. Based on enumeration results, flocks were categorized as Campylobacter negative or Campylobacter positive. The schedule of processing included the testing of Campylobacter-positive flocks before or after the testing of Campylobacter-negative flocks. During processing, flocks were also sampled for Campylobacter spp. before and after chilling. Campylobacter strains were identified with multiplex PCR and analyzed for relatedness with pulsed-field gel electrophoresis. Our results show that Campylobacter-negative flocks were indeed contaminated with Campylobacter strains originating from previously processed Campylobacter-positive flocks. Campylobacter isolates collected from carcasses originating from different farms processed on the same day showed similar pulsed-field gel electrophoresis patterns, confirming cross-contamination. These findings suggest that a simple logistic processing schedule can preserve the Campylobacter-negative status of broiler carcasses and result in products with enhanced food safety.

Sources and Spread of Thermophilic Campylobacter spp. during Partial Depopulation of Broiler Chicken Flocks

2008 ◽  
Vol 71 (2) ◽  
pp. 264-270 ◽  
Author(s):  
V. M. ALLEN ◽  
H. WEAVER ◽  
A. M. RIDLEY ◽  
J. A. HARRIS ◽  
M. SHARMA ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Broiler Chicken ◽  
Pulsed Field Gel Electrophoresis ◽  
The United Kingdom ◽  
Thermophilic Campylobacter ◽  
Broiler Flock ◽  
Commercial Broiler ◽  
Cleaning And Disinfection ◽  
Thinning Process ◽  
Campylobacter Spp

The practice of partial depopulation or thinning (early removal of a portion of birds from a commercial broiler flock) is a reported risk factor for Campylobacter colonization of residual birds because of the difficulty in maintaining biosecurity during the thinning process. The effect of this practice was studied in detail for 51 target flocks, each at a different growing farm belonging to one of seven major poultry companies throughout the United Kingdom. On 21 of these farms, the target flock was already colonized by Campylobacter, and at slaughter all cecal samples examined were positive, with a mean of 8 log CFU/g. An additional 27 flocks became positive within 2 to 6 days of the start of thinning and had similarly high levels of cecal carriage at slaughter. Just before the thinning process, Campylobacter was isolated frequently from the farm driveways, transport vehicles, equipment, and personnel. Strains from seven farms on which flocks became colonized after thinning were examined by pulsed-field gel electrophoresis typing. An association was found between strains occurring at specific sampling sites and those isolated subsequently from the thinned flocks. The results indicated that particular strains had spread from one farm to another when the farms were jointly owned by the same company and employed the same bird-catching teams and/or vehicles. These results highlight the need for better hygiene control in relation to catching equipment and personnel and more effective cleaning and disinfection of vehicles and bird-transport crates.

Efficacy of Mini VIDAS for the Detection of Campylobacter spp. from Retail Broiler Meat Enriched in Bolton Broth, with or without the Supplementation of Blood

2009 ◽  
Vol 72 (11) ◽  
pp. 2428-2432 ◽  
Author(s):  
LIN LIU ◽  
SYEDA K. HUSSAIN ◽  
ROBERT S. MILLER ◽  
OMAR A. OYARZABAL
Keyword(s):  
Gel Electrophoresis ◽  
Agar Plate ◽  
Meat Products ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Analysis ◽  
False Negatives ◽  
Broiler Meat ◽  
Automated Immunoassay ◽  
Campylobacter Spp ◽  
Pfge Profiles

The goals of this study were to evaluate the efficacy of the mini VIDAS automated immunoassay chemistry system to detect Campylobacter spp. from retail broiler meat enriched in Bolton broth supplemented with lysed blood (B+B) or without blood (B–B), and to detect positive samples at 24 versus 48 h after enrichment. Retail broiler meat was enriched and tested for Campylobacter spp. with the mini VIDAS and with an agar plate. Isolates were speciated with a multiplex PCR and typed with pulsed-field gel electrophoresis (PFGE) to evaluate relatedness of isolates collected from subsamples enriched in B+B or B–B. The number of Campylobacter-positive samples by mini VIDAS was similar (P > 0.05) to the results found with traditional plating media for naturally contaminated broiler meat, regardless of whether the comparison was made between B+B and B–B, or among different meat products (breast, tenders, and thighs). More positive samples were found at 48 h of enrichment than at 24 h of enrichment (P < 0.05). A Campylobacter jejuni:Campylobacter coli ratio of 4:1 was found in this study. Most of the isolates from both subsamples (B+B and B–B) were similar or identical by PFGE analysis, except for a few samples in which the PFGE profiles of the isolates from the subsamples were different. Mini VIDAS allowed for the detection of Campylobacter spp. within 48 h after enrichment. However, the sensitivity is similar to plate media, and retail broiler samples need to be enriched for 48 h to avoid false negatives.

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