Prevalence and characterization of Campylobacter jejuni from chicken meat sold in French retail outlets
Introduction
Campylobacter jejuni is the major cause of bacterial gastroenteritis in developed countries. Over 214,000 human cases were reported in 2012 in the European Union (EFSA and ECDC, 2014). Broiler chickens are generally recognized as the major source of contamination and the handling, preparation and consumption of broiler meat may account for 20 to 30% of human campylobacteriosis cases (EFSA, 2010b).
Campylobacteriosis is usually a self-limiting disease lasting only a few days. Treatment with antimicrobials is not usually required except in severe cases or immunocompromised patients. Macrolides, such as erythromycin are now generally considered to be the optimal drug for treating campylobacteriosis, with fluoroquinolones as an alternative therapy (Allos, 2001). However, the prevalence of antimicrobial resistant Campylobacter strains in humans is increasing especially resistance to fluoroquinolones and to a lesser extent, macrolides (Cody et al., 2010, EFSA (European Food Safety Authority) and ECDC (European Center for Disease Prevention and Control), 2011). Multiple resistance patterns to several classes of antibiotics are also emerging. Trends in microbial resistance have shown a clear link between use of antibiotics in the poultry industry and resistant isolates of Campylobacter in humans (Moore et al., 2006). These resistant zoonotic bacteria are of special concern since they might compromise the effective treatment of infections in humans.
C. jejuni is also a recognized antecedent cause of post-infectious neuropathies such as Guillain–Barré syndrome (GBS) or Miller–Fischer syndrome (MFS) (Nachamkin et al., 1998). Among the numerous factors involved in the development of such autoimmune diseases, molecular mimicry between the lipooligosaccharide (LOS) structures present on the cell surface of C. jejuni and gangliosides in peripheral nerves play a crucial role (Ang et al., 2004, Ang et al., 2002, Godschalk et al., 2004).
Nineteen distinct classes of LOS biosynthesis loci have been described based on major genetic differences, gene content and organization (Parker et al., 2008). Five of these classes (A, B, C, M and R) harbor sialyltransferase genes involved in incorporating sialic acids into the LOS (Gilbert et al., 2002, Parker et al., 2008, Parker et al., 2005). Sialylation of the LOS core has been associated with ganglioside mimicry and it has been shown that in GBS or MFS related to Campylobacter strains, the majority of strains express the LOS gene cluster A or B (Godschalk et al., 2004, Islam et al., 2009, Koga et al., 2006).
Moreover, C. jejuni expressing sialylated LOS (classes A, B and C) appear to be more invasive for intestinal cells in vitro than those expressing non-sialylated LOS (classes D and E) (Habib et al., 2009a, Louwen et al., 2008). Sialylation of the LOS was also associated with increased severity of enteric disease by Mortensen et al. (2009) but these results were not recently supported by Ellström et al. (2013).
Molecular typing is an important tool for elucidating the diversity and transmission routes of Campylobacter isolates contaminating the food chain. Various molecular typing methods are currently available for studying the population structure of Campylobacter (Wassenaar and Newell, 2000). These genotypic methods have shown that Campylobacter has marked genomic diversity and a slightly clonal population structure, in which the different lineages and the relatedness between isolates cannot be easily determined, particularly within the framework of long-term epidemiological studies. Based on a concept similar to multilocus enzyme electrophoresis, multilocus sequence typing (MLST) exploits the relative conservation in sequence of certain genes in which variations are more likely to be selectively neutral because of their housekeeping functions (Maiden et al., 1998). This approach was successfully developed to study Campylobacter populations (Dingle et al., 2001) and is now recognized as the gold standard typing method for this bacterial genus (Sheppard et al., 2009).
In France, little is known about the prevalence of pathogenic Campylobacter in chicken and their molecular characteristics. No population-based surveys have been conducted to investigate the molecular epidemiology of C. jejuni in chicken meat at points close to human consumption. In this study, a survey was carried out to investigate the prevalence of Campylobacter in the chicken meat supply chain. C. jejuni isolates were further characterized as in France, more than 80% of campylobacteriosis are due to this species. Thus MLST was used to determine the diversity and clonal relationships among the 175 C. jejuni isolates collected. In addition, we correlated the LOS classes assigned by PCR and antimicrobial resistance of the isolates with the genotypes assigned by MLST.
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Sampling of the monitoring plan
The monitoring plan concerned three types of chicken meat products: 120 carcasses, 121 chicken legs and 120 chicken fillets collected in retail outlets from April to December 2009. The plan was carried out in geographic areas representing the most significant consumption patterns in France (Anonymous, 2010). Chicken meat products were analyzed at the ANSES Laboratory of Ploufragan–Plouzané which is the French National Reference Laboratory for Campylobacter.
Bacteriological analysis
Enumeration and sample enrichment were
Prevalence and enumeration of Campylobacter on French chicken meat products
Campylobacter was detected on 76% (275/361) of the chicken products. As shown in Table 2, products with skin were significantly (p = 0.00) more heavily contaminated than products without skin, with a prevalence and an enumeration mean of Campylobacter of 90% (108/120) and 1.90 log10 CFU/g respectively on the carcasses, 85% (103/121) and 1.72 log10 CFU/g on the legs and finally 53% (64/120) and 0.82 log10 CFU/g on the fillets. The species found in the isolates collected from positive samples were
Discussion
Our study reports the prevalence and the population structure of C. jejuni in raw chicken meat retailed in France. Chicken meat products were collected in retail outlets from the ten most populated French departments, and are representative of chicken consumption in France. The results obtained therefore represent a realistic picture of the C. jejuni population in retail chicken meat available to consumers following processing. The prevalence of Campylobacter in French retail outlet is high,
Acknowledgments
We would like to thank the Direction Générale de l'Alimentation and the Conseil Général des Côtes d'Armor for financial support.
References (47)
- et al.
Ciprofloxacin-resistant campylobacteriosis in the UK
Lancet
(2010) - et al.
Characterization of clinical Campylobacter jejuni isolates with special emphasis on lipooligosaccharide locus class, putative virulence factors and host response
Int. J. Med. Microbiol.
(2013) - et al.
Severe diarrhoea caused by highly ciprofloxacin-susceptible Campylobacter isolates
Clin. Microbiol. Infect.
(2009) - et al.
The genetic bases for the variation in the lipo-oligosaccharide of the mucosal pathogen, Campylobacter jejuni. Biosynthesis of sialylated ganglioside mimics in the core oligosaccharide
J. Biol. Chem.
(2002) - et al.
Prevalence of potentially neuropathic Campylobacter jejuni strains on commercial broiler chicken products
Int. J. Food Microbiol.
(2011) - et al.
The epidemiology of antibiotic resistance in Campylobacter
Microbes Infect.
(2006) - et al.
Sialylation of Campylobacter jejuni lipo-oligosaccharides is associated with severe gastro-enteritis and reactive arthritis
Microbes Infect.
(2009) - et al.
Re-analysis of the risks attributed to ciprofloxacin-resistant Campylobacter jejuni infections
Int. J. Antimicrob. Agents
(2007) Campylobacter jejuni infections: update on emerging issues and trends
Clin. Infect. Dis.
(2001)- et al.
Ganglioside mimicry of Campylobacter jejuni lipopolysaccharides determines antiganglioside specificity in rabbits
Infect. Immun.
(2002)
The Guillain–Barre syndrome: a true case of molecular mimicry
Trends Immunol.
Microbiology of Food and Animal Feeding Stuffs — Horizontal Method for Detection and Enumeration of Campylobacter spp.
Résultats du plan de surveillance 2009 de la contamination par Salmonella et Campylobacter des viandes fraîches de poulet au stade de la distribution
Note de service DGAl/SDSSA/N2010-8211
A longitudinal 6-year study of the molecular epidemiology of clinical Campylobacter isolates in Oxfordshire, United Kingdom
J. Clin. Microbiol.
Decreasing trend of overlapping multilocus sequence types between human and chicken Campylobacter jejuni isolates over a decade in Finland
Appl. Environ. Microbiol.
Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli
Lett. Appl. Microbiol.
Antimicrobial resistance in Campylobacter strains isolated from French broilers before and after antimicrobial growth promoter bans
J. Antimicrob. Chemother.
Multilocus sequence typing system for Campylobacter jejuni
J. Clin. Microbiol.
Molecular characterization of Campylobacter jejuni clones: a basis for epidemiologic investigation
Emerg. Infect. Dis.
Analysis of the baseline survey on the prevalence of Campylobacter in broiler batches and of Campylobacter and Salmonella on broiler carcasses, in the EU, 2008. Part A: Campylobacter and Salmonella prevalence estimates
EFSA J.
EFSA Panel on Biological Hazards (BIOHAZ); scientific opinion on quantification of the risk posed by broiler meat to human campylobacteriosis in the EU
EFSA J.
The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in the European Union in 2009
EFSA J.
The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2010
EFSA J.
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These authors contributed equally to this paper.