Multilocus Sequence Typing, Pulsed-Field Gel Electrophoresis, and fla Short Variable Region Typing of Clonal Complexes of Campylobacter jejuni Strains of Human, Bovine, and Poultry Origins in Luxembourg

ABSTRACT Campylobacter jejuni is the most common cause of bacterial gastroenteritis in Luxembourg, with a marked seasonal peak during summer. The majority of these infections are thought to be sporadic, and the relative contribution of potential sources and reservoirs is still poorly understood. We monitored human cases from June to September 2006 (n = 124) by molecular characterization of isolates with the aim of rapidly detecting temporally related cases. In addition, isolates from poultry meat (n = 36) and cattle cecal contents (n = 48) were genotyped for comparison and identification of common clusters between veterinary and human C. jejuni populations. A total of 208 isolates were typed by sequencing the fla short variable region, macrorestriction analysis resolved by pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing (MLST). We observed a high diversity of human strains during a given summer season. Poultry and human isolates had a higher diversity of sequence types than isolates of bovine origin, for which clonal complexes CC21 (41.6%) and CC61 (18.7%) were predominant. CC21 was also the most common complex found among human isolates (21.8%). The substantial concordance between PFGE and MLST results for this last group of strains suggests that they are clonally related. Our study indicates that while poultry remains an important source, cattle could be an underestimated reservoir of human C. jejuni cases. Transmission mechanisms of cattle-specific strains warrant further investigation.

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Distribution of Campylobacter jejuni isolates from Turkey Farms and Different Stages at Slaughter Using Pulsed-Field Gel Electrophoresis and flaA-Short Variable Region Sequencing

Zoonoses and Public Health ◽

10.1111/j.1863-2378.2010.01383.x ◽

2011 ◽

Vol 58 (6) ◽

pp. 388-398 ◽

Cited By ~ 8

Author(s):

P. Perko-Mäkelä ◽

T. Alter ◽

P. Isohanni ◽

S. Zimmermann ◽

U. Lyhs

Keyword(s):

Campylobacter Jejuni ◽

Gel Electrophoresis ◽

Variable Region ◽

Pulsed Field Gel Electrophoresis ◽

Pulsed Field ◽

Short Variable Region

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Tracing Campylobacter jejuni strains along the poultry meat production chain from farm to retail by pulsed-field gel electrophoresis, and the antimicrobial resistance of isolates

Food Microbiology ◽

10.1016/j.fm.2012.04.020 ◽

2012 ◽

Vol 32 (1) ◽

pp. 124-128 ◽

Cited By ~ 41

Author(s):

Beatriz Melero ◽

Pekka Juntunen ◽

Marja-Liisa Hänninen ◽

Isabel Jaime ◽

Jordi Rovira

Keyword(s):

Antimicrobial Resistance ◽

Campylobacter Jejuni ◽

Gel Electrophoresis ◽

Pulsed Field Gel Electrophoresis ◽

Poultry Meat ◽

Meat Production ◽

Production Chain ◽

Pulsed Field

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A pulsed-field gel electrophoresis study of the genetic diversity of Campylobacter jejuni and Campylobacter coli in poultry flocks in Slovenia

Acta Veterinaria Hungarica ◽

10.1556/avet.58.2010.1.2 ◽

2010 ◽

Vol 58 (1) ◽

pp. 19-28 ◽

Cited By ~ 5

Author(s):

Igor Gruntar ◽

Matjaž Ocepek ◽

Jana Avberšek ◽

Jasna Mićunović ◽

Mateja Pate

Keyword(s):

Genetic Diversity ◽

Campylobacter Jejuni ◽

Gel Electrophoresis ◽

Pulsed Field Gel Electrophoresis ◽

Poultry Meat ◽

Campylobacter Coli ◽

Enteric Infection ◽

Pulsed Field ◽

Industrialised Countries ◽

Small Clusters

Campylobacter jejuniandC. colihave recently become the most frequent cause of bacterial foodborne enteric infection in most industrialised countries. Consumption and handling of undercooked contaminated poultry meat was identified as an important risk factor for human campylobacteriosis. The aim of this study was to ascertain the genetic diversity ofC. jejuniandC. colistrains isolated from poultry in Slovenia. A total of 68 isolates (42C. jejuni, 26C. coli) from faeces (n = 48), meat (n = 15) and skin/carcasses (n = 5) of chicken (n = 60) and turkey samples (n = 5) were analysed by pulsed-field gel electrophoresis.SmaI macrorestriction discriminated betweenC. jejuniandC. coliisolates.C. jejuniisolates exhibited a higher degree of diversity compared toC. coliisolates. In theC. jejunigroup, a number of small clusters were apparent, whileC. colistrains formed less but larger clusters. AdditionalKpnI digestion of selected isolates resulted in poor subtyping. Strains with identical or very similar profiles were found on different farms, either in the same or different regions and time periods. Some of the clones indicated possible cross-contamination at slaughterhouses.

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Typing of Campylobacter jejuni Isolates from Dogs by Use of Multilocus Sequence Typing and Pulsed-Field Gel Electrophoresis

Journal of Clinical Microbiology ◽

10.1128/jcm.01046-09 ◽

2009 ◽

Vol 47 (11) ◽

pp. 3466-3471 ◽

Cited By ~ 22

Author(s):

B. N. Parsons ◽

A. J. Cody ◽

C. J. Porter ◽

J. H. Stavisky ◽

J. L. Smith ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Gel Electrophoresis ◽

Multilocus Sequence Typing ◽

Pulsed Field Gel Electrophoresis ◽

Pulsed Field

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Comparison of PCR Binary Typing (P-BIT), a New Approach to Epidemiological Subtyping of Campylobacter jejuni, with Serotyping, Pulsed-Field Gel Electrophoresis, and Multilocus Sequence Typing Methods

Applied and Environmental Microbiology ◽

10.1128/aem.02215-09 ◽

2009 ◽

Vol 76 (5) ◽

pp. 1533-1544 ◽

Cited By ~ 33

Author(s):

Angela J. Cornelius ◽

Brent Gilpin ◽

Philip Carter ◽

Carolyn Nicol ◽

Stephen L. W. On

Keyword(s):

Campylobacter Jejuni ◽

Molecular Typing ◽

Gel Electrophoresis ◽

Multilocus Sequence Typing ◽

Diversity Index ◽

Pulsed Field Gel Electrophoresis ◽

Campylobacter Coli ◽

Pulsed Field ◽

Typing Methods ◽

Campylobacter Lari

ABSTRACT To overcome some of the deficiencies with current molecular typing schema for Campylobacter spp., we developed a prototype PCR binary typing (P-BIT) approach. We investigated the distribution of 68 gene targets in 58 Campylobacter jejuni strains, one Campylobacter lari strain, and two Campylobacter coli strains for this purpose. Gene targets were selected on the basis of distribution in multiple genomes or plasmids, and known or putative status as an epidemicity factor. Strains were examined with Penner serotyping, pulsed-field gel electrophoresis (PFGE; using SmaI and KpnI enzymes), and multilocus sequence typing (MLST) approaches for comparison. P-BIT provided 100% typeability for strains and gave a diversity index of 98.5%, compared with 97.0% for SmaI PFGE, 99.4% for KpnI PFGE, 96.1% for MLST, and 92.8% for serotyping. Numerical analysis of the P-BIT data clearly distinguished strains of the three Campylobacter species examined and correlated somewhat with MLST clonal complex assignations and with previous classifications of “high” and “low” risk. We identified 18 gene targets that conferred the same level of discrimination as the 68 initially examined. We conclude that P-BIT is a useful approach for subtyping, offering advantages of speed, cost, and potential for strain risk ranking unavailable from current molecular typing schema for Campylobacter spp.

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Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulsed-Field Gel Electrophoresis, Multilocus Sequence Typing (MLST), and Repetitive Sequence Polymerase Chain Reaction (rep-PCR) Reveals Different Discriminatory Capabilities

Microbial Ecology ◽

10.1007/s00248-009-9571-3 ◽

2009 ◽

Vol 58 (4) ◽

pp. 843-855 ◽

Cited By ~ 23

Author(s):

Melissa K. Wilson ◽

Alison B. Lane ◽

Bibiana F. Law ◽

William G. Miller ◽

Lynn A. Joens ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Campylobacter Jejuni ◽

Repetitive Sequence ◽

Gel Electrophoresis ◽

Multilocus Sequence Typing ◽

Pulsed Field Gel Electrophoresis ◽

Chain Reaction ◽

Pulsed Field ◽

Pan Genome ◽

Polymerase Chain

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Phenotypic and Molecular Analysis of Penicillin-Nonsusceptible Streptococcus pneumoniae Isolates in Poland

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01072-06 ◽

2006 ◽

Vol 51 (1) ◽

pp. 40-47 ◽

Cited By ~ 13

Author(s):

Ewa Sadowy ◽

Radosław Izdebski ◽

Anna Skoczyńska ◽

Paweł Grzesiowski ◽

Marek Gniadkowski ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Molecular Analysis ◽

Gel Electrophoresis ◽

Multilocus Sequence Typing ◽

Susceptibility Testing ◽

Bacterial Pathogen ◽

Pulsed Field Gel Electrophoresis ◽

Pulsed Field ◽

Related Clone ◽

Sequence Types

ABSTRACT β-Lactams are the drugs of choice for the treatment of infections caused by the important bacterial pathogen Streptococcus pneumoniae. The recent growth of resistance of this organism to penicillin observed worldwide is of the highest concern. In this study, using 887 surveillance pneumococcal isolates recovered in Poland from 1998 to 2002, we observed the increase in penicillin nonsusceptibility from 8.7% to 20.3%. All of the 109 penicillin-nonsusceptible S. pneumoniae (PNSP) isolates identified, together with 22 archival PNSP isolates from 1995 to 1997, were subsequently analyzed by susceptibility testing, serotyping, profiling of pbp genes, pulsed-field gel electrophoresis, and multilocus sequence typing (MLST). Four predominant serotypes, serotypes 6B, 9V, 14, and 23F, characterized 85.5% of the isolates. MLST revealed the presence of 34 sequence types, 15 of which were novel types. Representatives of seven multiresistant international clones (Spain23F-1, Spain6B-2, Spain9V-3, Taiwan23F-15, Poland23F-16, Poland6B-20, and Sweden15A-25) or their closely related variants comprised the majority of the study isolates. The spread of Spain9V-3 and its related clone of serotype 14/ST143 has remarkably contributed to the recent increase in penicillin resistance in pneumococci in the country.

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