Determining the DNA Fingerprinting Profiles of Salmonella spp. Isolated from Raw Poultry Meats and Human Clinical Samples from the Same Geographic Area Using Pulsed Field Gel Electrophoresis.

2021 ◽  
Author(s):  
Eyob Mazengia ◽  
John Scott Meschke ◽  
Kay Greeson ◽  
Shaohua Zhao ◽  
Jason Abbott ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Sampling Period ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Isolates ◽  
Pulsed Field ◽  
Poultry Products ◽  
Poultry Processing ◽  
One Year ◽  
Raw Poultry ◽  
Pfge Profiles

Foods of animal origin, such as poultry, eggs, and pork, are recognized sources of infection with Salmonella , but determining the proportion of foodborne infections resulting from different food sources has been challenging. In the current study, 141 Salmonella strains isolated from poultry products (n=1322) purchased over a period of one year from retail stores across Seattle, Washington, were subtyped by Pulsed Field Gel Electrophoresis (PFGE) using restriction enzyme Xba I. The objectives of the study were: I) to analyze the longitudinal distribution of Salmonella PFGE profiles throughout the sampling period and their clonality within and between different poultry processing establishments; II) to determine the association between PFGE profiles of Salmonella isolated from locally distributed poultry products to those of clinical isolates submitted to the Washington State Department of Health Laboratories (WA-DOH); and III) to compare the PFGE profiles of Salmonella isolated from the National Antimicrobial Resistance Monitoring System (NARMS) retail meats program. Over the sampling period of one year, multiple indistinguishable PFGE patterns were found across multiple poultry processing establishments. There were 12 out of 30 unique PFGE profiles of Salmonella isolated from locally purchased poultry products that were indistinguishable from PFGE profiles of Salmonella from clinical isolates submitted to the WA-DOH. When the PFGE profiles from the poultry samples were compared to those found in NARMS’ database, there were eight indistinguishable PFGE matches isolated from chicken breasts, ground turkey, and ground beef from multiple states. While this study found some association between PFGE profiles from raw poultry products and those of clinical isolates from the same geographical area, it does not prove that all of those clinical isolates were infections that resulted from the consumption or handling of poultry.

scholarly journals Use of Pulsed-Field Gel Electrophoresis To Characterize the Heterogeneity and Clonality of Salmonella Isolates Obtained from the Carcasses and Feces of Swine at Slaughter

2003 ◽  
Vol 69 (7) ◽  
pp. 4177-4182 ◽  
Author(s):  
Laura Wonderling ◽  
Rachel Pearce ◽  
F. Morgan Wallace ◽  
Jeffrey E. Call ◽  
Ingrid Feder ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Sampling Period ◽  
Pulsed Field Gel Electrophoresis ◽  
Processing Plant ◽  
Pulsed Field ◽  
Software Analysis ◽  
Plant Environment ◽  
Swine Feces ◽  
Matched Samples ◽  
Pfge Profiles

ABSTRACT Salmonella enterica isolates were recovered from swine at a collaborating processing plant over a 2-month period in the spring of 2000. In the present study, molecular subtyping by pulsed-field gel electrophoresis (PFGE) was performed on the 581 confirmed Salmonella isolates from the 84 Salmonella-positive samples obtained from the previous study. A total of 32 different PFGE pulsotypes were observed visually, and a BioNumerics software analysis clustered those pulsotypes into 12 PFGE groups. The B, F, and G groups predominated throughout the sampling period and were isolated from 39, 22, and 13% of the swine, respectively. In addition, multiple isolates were obtained from 67 of the 84 Salmonella-positive samples, and subtyping revealed multiple PFGE profiles in 35 of these 67 (62%) samples. Both carcass and fecal isolates of Salmonella were recovered from 13 swine, resulting in “matched” samples. Molecular typing of the 252 isolates recovered from the matched samples revealed that 7 (54%) of the 13 carcasses were contaminated with Salmonella pulsotypes that were not isolated from the feces of the same animal. Conversely, from 6 of the 13 (46%) matched animals, Salmonella clonal types were isolated from the feces that were not isolated from the carcass of the same animal. These data establish that each lot of swine introduces new contaminants into the plant environment and that swine feces from one animal can contaminate many carcasses. In addition, these results indicate that the examination of multiple Salmonella isolates from positive samples is necessary to determine the variety of potential contaminants of swine carcasses during slaughter and processing.

Isolation and Pulsed-Field Gel Electrophoresis Typing of Listeria monocytogenes from Modified Atmosphere Packaged Fresh-Cut Vegetables Collected in Ireland

2006 ◽  
Vol 69 (10) ◽  
pp. 2524-2528 ◽  
Author(s):  
GILLIAN A. FRANCIS ◽  
DAVID O'BEIRNE
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Fruits And Vegetables ◽  
Pulsed Field Gel Electrophoresis ◽  
Leafy Vegetables ◽  
Modified Atmosphere ◽  
Pulsed Field ◽  
Fruit Samples ◽  
Fresh Cut ◽  
Pfge Profiles

The incidence of Listeria monocytogenes in modified atmosphere packaged fresh-cut fruits and vegetables from chill cabinets of a supermarket in Ireland was investigated over a 2-year period. Overall, 9.58% of fresh-cut produce was contaminated with Listeria spp. Various species of Listeria were isolated from samples, including L. monocytogenes, L. seeligeri, L. innocua, L. welshimeri, and L. ivanovii. No fruit samples contained detectable L. monocytogenes. Overall, a total of 21 L. monocytogenes isolates (2.9% of samples) were recovered from a range of products, including dry coleslaw mix (80% shredded cabbage and 20% shredded carrot), bean sprouts, and leafy vegetables such iceberg, romaine, and radicchio lettuce and mixed salad leaves (curly endive, escarole, and radicchio leaves). Dry coleslaw mix appeared to have the highest incidence of Listeria contamination (20%) compared with other products. Listeria contamination was more frequent (P < 0.05) during the summer and autumn months than during the winter and spring months. The 21 L. monocytogenes isolates were subsequently subtyped by genomic macrorestriction techniques using ApaI with pulsed-field gel electrophoresis (PFGE). PFGE of digested DNA produced bands of 79 to 518 kb. Four PFGE profiles were identified, and approximately 50% of the isolates were associated with profile 1. This study indicates that fresh-cut vegetables packaged under a modified atmosphere can support growth of numerous species of Listeria, including L. monocytogenes.

scholarly journals Molecular typing of Escherichia coli O157[ratio ]H7 (H−) isolates from cattle in Japan

1999 ◽  
Vol 122 (2) ◽  
pp. 337-341 ◽  
Author(s):  
M. AKIBA ◽  
T. MASUDA ◽  
T. SAMESHIMA ◽  
K. KATSUDA ◽  
M. NAKAZAWA
Keyword(s):  
Escherichia Coli ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Outbreak Strain ◽  
Pulsed Field ◽  
E Coli ◽  
Biological Methods ◽  
Pfge Profiles

A total of 77 Escherichia coli O157[ratio ]H7 (H−) isolates from cattle in Japan were investigated by molecular biological methods. Most of these isolates (43 isolates) possessed the stx2 gene, but not stx1. Fifteen bacteriophage types and 50 pulsed-field gel electrophoresis (PFGE) profiles were observed. One isolate was indistinguishable from the human outbreak strain by these methods. This indicates that cattle must be considered as a possible source of human E. coli O157[ratio ]H7 infection in Japan.

scholarly journals Molecular epidemiology of nalidixic acid-resistant campylobacter isolates from humans and poultry by pulsed-field gel electrophoresis and flagellin gene analysis

2002 ◽  
Vol 129 (1) ◽  
pp. 227-231 ◽  
Author(s):  
T. L. WU ◽  
L. H. SU ◽  
J. H. CHIA ◽  
T. M. KAO ◽  
C. H. CHIU ◽  
...  
Keyword(s):  
Nalidixic Acid ◽  
Gel Electrophoresis ◽  
Antimicrobial Agents ◽  
Fragment Length Polymorphism ◽  
Pulsed Field Gel Electrophoresis ◽  
Flagellin Gene ◽  
Pulsed Field ◽  
Poultry Products ◽  
Typing Methods ◽  
Human Infections

To investigate the potential of poultry products as the source of human infections associated with quinolone-resistant campylobacters, 140 human and 75 poultry isolates of nalidixic acid-resistant campylobacters were collected between 1996 and 1998, and analysed by two molecular typing methods. By the analysis of restriction fragment length polymorphism of the flagellin gene, 33 distinct patterns were obtained, with 18 of which shared by both human (89%) and poultry (93%) isolates. By the pulsed-field gel electrophoresis of SmaI-restricted macrofragments, 105 different profiles were obtained, and 11 were found in both human (40%) and poultry (23%) isolates. When the two typing methods were combined, 112 unique genotypes were obtained, 11 of which were shared by both populations, including 53 (38%) human isolates and 14 (19%) poultry isolates. Although domestic poultry products are still important sources of the quinolone-resistant campylobacter infections in humans, there are other factors that might contribute to these increasing infections simultaneously. A more stringent policy in the use of antimicrobial agents in food animals can no longer be ignored.

scholarly journals Intermediately virulent Rhodococcus equi isolates from pigs in Slovenia: discovery of new plasmid types and assessment of genetic diversity by pulsed-field gel electrophoresis

2009 ◽  
Vol 54 (No. 3) ◽  
pp. 111-117 ◽  
Author(s):  
M. Pate ◽  
M. Ocepek ◽  
I. Zdovc ◽  
C. Minato ◽  
Y. Ohtsu ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Gel Electrophoresis ◽  
Detailed Comparison ◽  
Rhodococcus Equi ◽  
Pulsed Field Gel Electrophoresis ◽  
High Genetic Diversity ◽  
Pulsed Field ◽  
Large Plasmids ◽  
Multiple Strains ◽  
Pfge Profiles

The presence of large plasmids in 30 <I>Rhodococcus equi</I> strains isolated from pig lymph nodes with granulomatous changes was investigated. Plasmid DNAs were isolated and digested with the restriction endonucleases <I>Bam</I>HI, <I>Eco</I>RI, <I>Eco</I>T22I and <I>Hind</I>III for detailed comparison and estimation of plasmid sizes. A total of nine isolates were identified as intermediately virulent (VapB-positive), harbouring large plasmids of type 5 (<I>n</I> = 5) and four new variants that we tentatively designated as type 19 (<I>n</I> = 1), 20 (<I>n</I> = 1), 21 (<I>n</I> = 1) and 24 (<I>n</I> = 1). All isolates were subjected to genotyping with pulsed-field gel electrophoresis (PFGE). High genetic diversity was observed: 21 distinct genotypes were detected; five were found in multiple isolates and the others were unique. Isolates of the same plasmid type exhibited different PFGE profiles and vice versa. In a few cases, multiple strains from certain farms were analysed, the majority of which exhibited diverse PFGE profiles. Our findings demonstrate the presence of a wide variety of <I>R. equi</I> strains even in small confined environments such as farms. This is the first molecular epidemiology study of intermediately virulent <I>R. equi</I> isolates from Slovenian pigs.

Comparison of Danish Isolates of Salmonella entericaSerovar Enteritidis PT9a and PT11 from Hedgehogs (Erinaceus europaeus) and Humans by Plasmid Profiling and Pulsed-Field Gel Electrophoresis

2000 ◽  
Vol 38 (10) ◽  
pp. 3631-3635 ◽  
Author(s):  
B. Nauerby ◽  
K. Pedersen ◽  
H. H. Dietz ◽  
M. Madsen
Keyword(s):  
Gel Electrophoresis ◽  
Fragment Length Polymorphism ◽  
Phage Type ◽  
Pulsed Field Gel Electrophoresis ◽  
Clonal Lineage ◽  
S1 Nuclease ◽  
Pulsed Field ◽  
Plasmid Profiling ◽  
Susceptibility Patterns ◽  
Pfge Profiles

During the years 1994 to 1998, 10 strains of Salmonella enterica serovar Enteritidis phage type 11 (PT11) and 6 PT9a strains were isolated from Danish hedgehogs, together with 7 strains that did not yield phage susceptibility patterns conforming with any known phage type (routine dilution no conformity [RDNC]). From 1995 to 1998, five Danish patients were reported infected with serovar Enteritidis PT11 and two with PT9a. All serovar Enteritidis PT11, PT9a, and RDNC isolates from hedgehogs and humans were analyzed by pulsed-field gel electrophoresis (PFGE), plasmid profiling, and restriction fragment length polymorphism (RFLP) of plasmids. By use of S1 nuclease and HindIII, the PT11 and PT9a isolates had identical plasmid profiles and RFLP patterns, which differed from the RDNC profiles. The PFGE profiles were identical for all serovar Enteritidis PT11 and PT9a strains from hedgehogs, four of five human strains of serovar Enteritidis PT11, and two human strains of serovar Enteritidis PT9a, irrespective of restriction enzyme, whereas the last human strain deviated slightly when NotI was used but not when XbaI or SpeI was used. The results indicate that serovar Enteritidis PT9a and PT11 are closely related and that PT11 and PT9a from Danish hedgehogs and humans belong to the same clonal lineage.

scholarly journals Pulsed-Field Gel Electrophoresis Profile Changes Resulting from Spontaneous Chromosomal Deletions in Enterohemorrhagic Escherichia coli O157:H7 during Passage in Cattle

2009 ◽  
Vol 75 (17) ◽  
pp. 5719-5726 ◽  
Author(s):  
Noriyo Yoshii ◽  
Yoshitoshi Ogura ◽  
Tetsuya Hayashi ◽  
Takashi Ajiro ◽  
Toshiya Sameshima ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Enterohemorrhagic Escherichia Coli ◽  
Sequencing Analysis ◽  
Pulsed Field ◽  
Chromosomal Deletions ◽  
Profile Changes ◽  
Dna Sequencing Analysis ◽  
Pfge Profiles

ABSTRACT A total of 905 enterohemorrhagic Escherichia coli (EHEC) O157:H7 isolates that were recovered from experimentally infected cattle, in addition to the inoculated strain, were analyzed by pulsed-field gel electrophoresis (PFGE). Twelve PFGE profiles other than that of the inoculated strain were observed. We successfully identified five distinct chromosomal deletions that affected the PFGE profiles using whole-genome PCR scanning and DNA sequencing analysis. The changes in PFGE profiles of EHEC O157:H7 isolates after passage through the intestinal tract of cattle were partially generated by deletion of chromosomal regions.

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