The viability ofCampylobacter jenuni on refrigerated chicken drumsticks

Radappertized chicken drumsticks were experimentally contaminated with suspensions ofCampylobacter jejuni in two trials. Qualitative analysis on drumsticks with an initial level of contamination of 4.8×10³ CFU/cm² showed that viability was retained for at least 10 days of storage at either 9° or –12°C.In a second quantitative trial, the level of contamination declined from 9.9×10² CFU/cm² to 4.5×10¹ CFU/cm² after 7 days at –20°C. Thereafter,C. jejuni persisted at levels ranging from 1.8×10¹ to 0.2×10¹ CFU/cm² through the 26th week of storage. Drumsticks held at 4°C showed a significant decline in count from 9.9×10² CFU/cm² to 1.8×10² CFU/cm² on day 7.It is concluded that the viability ofC. jejuni on chicken parts is maintained under both refrigerated and freezing conditions which approximate commercial storage. This is of significance to the meat industry and consumers.     

Campylobacter jejuni contamination of eggs

Contamination of commercial table eggs with a fecal suspension containing 4.4×10⁶ CFU/gCampylobacter jejuni resulted in shell penetration in 3/70 eggs and recovery of the organism from homogenized egg contents in 1/70 eggs. Viability ofC. jejuni on the shell surface was retained for only 16 hours, attributed to desiccation of the fecal suspension. A field survey of three commercial laying farms and their associated egg-packing plants showed that hens demonstrated to be fecal shedders ofC. jejuni (12% to 62% incidence) did not produce infected eggs. The organism could not be detected in the environment of the packing plant, including grading machinery and effluent.     

The efficacy of selected probiotic and prebiotic combinations in reducing Campylobacter colonization in broiler chickens

Campylobacter is one of the leading causes of foodborne illness worldwide and is associated with consumption of poultry products. One potential strategy to reduce Campylobacter colonization in poultry is the use of probiotics. In an attempt to reduce Campylobacter in poultry, we isolated 117 GRAS (generally recognized as safe) bacterial isolates from the ceca of healthy birds and evaluated their efficacy against Campylobacter, using a soft agar overlay technique. Twenty-six isolates (Bacillus and Lactobacillus spp.) with greatest ability to inhibit Campylobacter in vitro, were tested in birds. In these studies, bacterial isolates were administered orally on day-of-hatch and, chicks were challenged with a four-strain mixture of wild type C. jejuni (∼10⁶ CFU/mL) on day 7. Cecal samples were collected on day 14 for Campylobacter enumeration. Out of 26 isolates, 3 isolates demonstrated a reduction in Campylobacter counts (1 to 2 logs). In an attempt to improve the efficacy of these 3 isolates, follow-up studies were conducted with various doses of 2 prebiotics (fructooligosaccharide [FOS] or mannan oligosaccharide [MOS]) in the feed. Campylobacter counts were not reduced for any of the FOS treatments (0.125%, 0.25%, or 0.5%) or the MOS treatments (0.04%, 0.08%, or 0.16%), but were reduced for the combination of isolate 3 with 0.04% MOS in feed. The results of this study indicate that the selection and administration of bacterial isolates in combination with selected prebiotics may reduce enteric Campylobacter counts in preharvest poultry.     

Combined Campylobacter jejuni and Campylobacter coli Rapid Testing and Molecular Epidemiology in Conventional Broiler Flocks

Campylobacter spp. are important causes of bacterial zoonosis, most often transmitted by contaminated poultry meat. From an epidemiological and risk assessment perspective, further knowledge should be obtained on Campylobacter prevalence and genotype distribution in primary production. Consequently, 15 Austrian broiler flocks were surveyed in summer for their thermophilic Campylobacter spp. contamination status. Chicken droppings, dust and drinking water samples were collected from each flock at three separate sampling periods. Isolates were confirmed by PCR and subtyped. We also compared three alternative methods (culture‐based enrichment in Bolton broth, culture‐independent real‐time PCR and a lateral‐flow test) for their applicability in chicken droppings. Twelve flocks were found to be positive for thermophilic Campylobacter spp. during the entire sampling period. Seven flocks (46.6%) were contaminated with both, C. jejuni and C. coli, five flocks harboured solely one species. We observed to a majority flock‐specific C. jejuni and C. coli genotypes, which dominated the respective flock. Flocks within a distance <2 km shared the same C. jejuni genotypes indicating a cross‐contamination event via the environment or personnel vectors. Multilocus sequence typing (MLST) of C. jejuni revealed that the majority of isolates were assigned to globally distributed clonal complexes or had a strong link to the human interface (CC ST‐446 and ST4373). The combination of techniques poses an advantage over risk assessment studies based on cultures alone, as, in the case of Campylobacter, occurrence of a high variety of genotypes might be present among a broiler flock. We suggest applying the lateral‐flow test under field conditions to identify ‘high‐shedding’ broiler flocks at the farm level. Consequently, poultry farmers and veterinarians could improve hygiene measurements and direct sanitation activities, especially during the thinning period. Ultimately, real‐time PCR could be applied to quantify Campylobacter spp. directly from chicken droppings and avoid non‐interpretable results achieved by culture‐dependent methods.     

Carry-over of thermophilic Campylobacter spp. between sequential and adjacent poultry flocks

Nineteen flocks of four poultry species were monitored at a veterinary field station to investigate the distribution and spread of Campylobacter genotypes between sequential and adjacent flocks. Caecal and liver samples were obtained at frequent intervals from birds of all flocks and examined for Campylobacter. Amplified fragment length polymorphism (AFLP) analysis was performed to genotype Campylobacter isolates. Of the 1643 caecal and liver samples investigated, 452 (27.5%) caecal samples and 11 (0.7%) liver samples contained Campylobacter. Of the caecal isolates 76.3% were identified as Campylobacter jejuni and 23.7% were identified as Campylobacter coli. Poultry flocks were largely colonized by more than one AFLP type and an intense exchange of Campylobacter genotypes between different poultry flocks occurred. These findings indicate that multiple genotypes can constitute the Campylobacter population within single poultry flocks, hinting to different sources of exposure and/or genetic drifts within the Campylobacter population. Nevertheless, in most flocks single Campylobacter genotypes predominated. Some strains superseded others resulting in colonization by successive Campylobacter genotypes during the observation period. In conclusion, the data demonstrate that the large genetic diversity of Campylobacter must be considered in epidemiological evaluations and microbial risk assessments of Campylobacter in poultry.     

Prevalence,Molecular Characterization and Antimicrobial Resistance of Thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> Isolates from Cattle,Hens, Broilers and Broiler Meat in South-eastern Italy

Eleven cattle farms, 8 layer farms, 7 broiler farms and 30 broiler meat samples were investigated in south-eastern Italy throughout 2003 to evaluate the prevalence, the molecular type and antimicrobial resistance of thermophilic Campylobacters. A total of 398 samples were analysed. One Campylobacter isolate for each positive faecal swab and three isolates per positive broiler meat sample were selected for further analysis. Multiplex PCR was performed for species-level identification and PCR-RFLP of the flagellin A gene for genotyping. Resistance to 14 antimicrobials was studied in 188 Campylobacter isolates. Prevalence of campylobacters was high both on farms (100%) and in food samples (73%). On 4/11 cattle farms and on 10/15 poultry farms more than one species was isolated. The presence of more than one genotype was found on 8/11 cattle farms, on 10/15 poultry farms and in 8/22 Campylobacter-positive food samples. High rates of resistance to quinolone were observed: 9/31 (29%) C. jejuni bovine isolates, 4/22 (18%) C. jejuni poultry isolates, and 14/26 (54%) C. coli poultry isolates. Resistance to sulphamethoxazole-trimethoprim was also observed frequently: 18/26 (69%) of the avian C. coli strains, 25/31 (80%) of the C. jejuni strains isolated from poultry and 15/22 (68%) of those isolated from cattle were resistant. There was a significant difference between the rate of resistance to macrolides of C. coli and C. jejuni isolated in poultry, which amounted to 23% and 3%, respectively. This study provided data on the prevalence and antimicrobial resistance of thermophilic campylobacters in south-eastern Italy and confirmed that flaA-typing is an efficient tool to study the epidemiology of Campylobacter strains in short-term investigations.     

Campylobacter spp. in Irish Feedlot Cattle: A Longitudinal Study Involving Pre‐harvest and Harvest Phases of the Food Chain

The aim of this study was to investigate faecal shedding and transmission of Campylobacter spp. in cohorts of cattle within a feedlot, to assess subsequent contamination of carcasses with this pathogen and to identify risk factors associated with faecal shedding of Campylobacter spp. A cohort of 133 heifers housed in four adjacent pens was examined over a five and a half month period, from entering the feedlot to slaughter. A parallel investigation of individual rectal faecal samples and pen environmental samples were taken at monthly intervals from November to February. The entire outer and inner surfaces of a carcass side of each animal were swabbed immediately following slaughter. Campylobacter spp. were isolated from 322 (54%) of the 600 rectal faecal samples. Campylobacter jejuni and C. coli accounted for 69 and 29.7% of the isolate recovered, respectively. A total of 159 environmental samples were examined, of these Campylobacter spp. was isolated from 46 samples (29%). Campylobacter jejuni and C. coli accounted for 35 and 59% of these isolates, respectively. Campylobacter spp. was not isolated from any of the dressed carcasses. Logistic regression indicated prevalence of Campylobacter spp. faecal shedding within pens was positively correlated to the pen, the month of sampling and the Campylobacter spp. contamination status of the pen dividing bars and the water trough surface. Campylobacter spp. should be considered as a pathogen shed in the faeces of a substantial proportion of feedlot cattle. However, with good hygienic practice during harvest, a very low level of this pathogen can be achieved on dressed carcasses.     

Simulation model for Campylobacter cross-contamination during poultry processing at slaughterhouses

Broiler meat is regarded as the most common source of Campylobacter infection and risk management measures are required to reduce broiler meat contamination. Among the quantitative risk assessments for Campylobacter in broiler meat, evaluation of the poultry processing stage is particularly important for predicting the contamination level of broiler meat and the effects of control measures. In this study, we built a simulation model for cross‐contamination during poultry processing focusing on Campylobacter contamination at the individual carcass level. Using this model, we examined changes in the prevalence of contaminated carcasses and the number of Campylobacter per carcass after processing. As a result, it was found that the prevalence and number of Campylobacter after processing were largely influenced by the number of Campylobacter on the contaminated carcasses before processing. In the baseline model, where it was assumed that the mean number of Campylobacter on contaminated carcasses before processing was 4.7 log₁₀ cfu per carcass, the prevalence after processing was less than that before processing. Although the median value of Campylobacter on contaminated carcasses was reduced after processing, the distributions after processing became wider and the upper limit of the 95% credible interval of Campylobacter on contaminated carcasses remained elevated. The individual‐based simulation model can trace individual level changes considering discrete interactions, while models tracing mean values cannot handle these interactions in detail. The individual‐based approach is considered useful for modelling cross‐contamination among individual carcasses during poultry processing.     

Antimicrobial Resistance Profiles of Campylobacter spp. Isolated from Broiler Chicken Meat of Estonian,Latvian and Lithuanian Origin at Estonian Retail Level and from Patients with Severe Enteric Infections in Estonia

The resistance patterns of Campylobacter spp. isolated from retail broiler chicken meat originating either from Estonia, Lithuania or Latvia collected in Estonia were determined. Additionally, in collaboration with the laboratories of several Estonian hospitals, antimicrobial susceptibility patterns were determined for Campylobacter isolates from patients with severe Campylobacter enteric infections. The isolates were identified at the species level by the PCR method. Respectively, 88.8% of the isolates were C. jejuni, and 11.2% were C. coli. In total, 126 Campylobacter isolates of broiler chicken meat and human origin were tested for minimal inhibitory concentrations (MICs) with the broth microdilution VetMIC^TH method (National Veterinary Institute; Uppsala, Sweden) for a total of six antimicrobials. Resistance to one or more antimicrobials was detected in 62 (63.3%) of Campylobacter broiler chicken meat isolates and in 20 (71.4%) of human‐origin isolates. Large proportions of the broiler chicken meat isolates were resistant to ciprofloxacin (60.2%). Multidrug resistance (i.e. to three or more unrelated antimicrobials) was detected in five (5.1%) C. jejuni isolates. Among the human isolates, 20 (71.4%) were resistant to fluoroquinolones, and two (7.1%) C. jejuni isolates exhibited multidrug resistance. The chicken meat isolates of Estonian origin were the most susceptible. However, a high proportion of fluoroquinolone‐resistant C. jejuni isolates were found in Latvian and Lithuanian products. The results of this study indicate that the problems caused by the inappropriate use of antimicrobials extend beyond the country in which a food originates; therefore, both domestic and international interventions and agreements are required to implement common policies on antimicrobial usage and to minimize the emergence of Campylobacter drug resistance.     

The isolation and characterization of Campylobacter jejuni bacteriophages from free range and indoor poultry

Six hundred and sixty one samples – primarily fresh chicken faeces – were processed to isolate wild type Campylobacter jejuni bacteriophages, via overlay agar methods using C. jejuni NCTC 12662. The aims of this study were to isolate and purify bacteriophages and then test for their ability to lyse field strains of C. jejuni in vitro. Of all samples processed, 130 were positive for bacteriophages. A distinct difference was observed between samples from different poultry enterprises. No bacteriophages could be isolated from indoor broilers. The majority of bacteriophages were isolated from free range poultry – both broilers and egg layers. Bacteriophages were purified and then selected for characterization based on their ability to produce clear lysis on plaque assay, as opposed to turbid plaques. Two hundred and forty one C. jejuni field isolates were tested for sensitivity to the bacteriophages. Lysis was graded subjectively and any minimal lysis was excluded. Using this system, 59.0% of the C. jejuni isolates showed significant sensitivity to at least one bacteriophage. The sensitivity to individual bacteriophages ranged from 10.0% to 32.5% of the C. jejuni isolates. Five bacteriophages were examined by electron microscopy and determined to belong to the Myoviridae family. The physical size, predicted genetic composition and genome size of the bacteriophages correlated well with other reported Campylobacter bacteriophages. The reasons for the observed difference between indoor broilers and free range poultry is unknown, but are postulated to be due to differences in the Campylobacter population in birds under different rearing conditions.     

Diversity of Campylobacter jejuni and Campylobacter coli Genotypes from Human and Animal Sources from Rio de Janeiro, Brazil

To compare the genotypes of Campylobacter jejuni and Campylobacter coli isolates of human and animal origin collected in Rio de Janeiro City, 30 C. jejuni and 35 C. coli isolates from animal sources (n = 45) and human patients with gastroenteritis (n = 20) were genotyped by PCR-based techniques, namely random amplified polymorphic DNA (RAPD-PCR) and enterobacterial repetitive intergenic consensus sequence (ERIC-PCR). RAPD-PCR identified 50 types and ERIC-PCR identified 22 genotypes, among the 65 Campylobacter isolates. Both PCR methods discriminated the C. jejuni and C. coli groups of isolates. Combining the results of both methods, no single genotype was shared between isolates from human and animal sources. Two groups of two C. coli isolates each with identical genotypes were found among poultry and pig isolates. A high level of genetic diversity observed among the Campylobacter isolates suggests lack of overlap between isolates from different sources.     

Molecular detection of Campylobacter species and Cytolethal distending toxin isolated from chicken livers in Tabriz

Introduction and purpose: Campylobacter jejuni and coli are zoonotic bacteria commonly associated With human diarrhea and usually transmit through consumption of meat and poultry contaminated products such as heart and liver. Cytolethal distending toxin (cdt) in Campylobacter spp. is among the significant virulence factors of these bacteria in the intestine. The purpose of this study was to determine the prevalence of Campylobacter spp. and presence of cdt genes among isolated bacteria. Materials and methods: In this cross sectional study, 100 chicken livers were examined. Detection, bacterial enumeration and isolation of Campylobacter spp. was done using Campylobacter selective agar media containing Campylobacter growth supplement, gram staining, catalase and oxidase production, hippurate hydrolysis and PCR molecular technique. Also the presence of cdt genes were detected using PCR assay. Results: From 100 studied liver samples, 43 were contaminated with Campylobacter spp. Among them 31(72 %) samples had Campylobacter jejuni and 12 (28 %) had Campylobacter coli. All Campylobacter jejuni isolates contained cdtA ،cdtB and cdtC genes. However, all of these genes detected in 9 (75 %) of isolated coli. Conclusion: The results of this study showed that great percentages of chicken livers in Tabriz were contaminated with Campylobacter.     

Detection and identification of food‐borne pathogens of the genera Campylobacter,Arcobacter and Helicobacter by multiplex PCR in poultry and poultry products

The objective of this study was to develop a multiplex polymerase chain reaction (PCR) to detect and differentiate food‐borne pathogens of the three genera Campylobacter, Arcobacter and Helicobacter in a single step procedure. One common reverse primer and three genus‐specific forward primers were designed by hybridizing to the 16S rRNA of selected reference strains. Besides the species with significance as food‐borne pathogens isolated from poultry meat –Campylobacter jejuni, Campylobacter coli, Arcobacter butzleri and Helicobacter pullorum– several other members of these genera were tested to determine the specificity of the designed multiplex PCR. In total, 20 ATCC and NCTC reference strains of Campyobacter, Arcobacter and Helicobacter were used to evaluate the PCR. Specific amplificates were obtained from all thermophilic species of Campylobacter as well as from species of Arcobacter and Helicobacter. No amplification product was obtained from the non‐thermophilic Campylobacter, C. hyointestinalis and C. fetus. Furthermore, a total of 43 field strains of the three genera isolated from poultry, pigs, cattle and humans were investigated using this PCR. To confirm the classification of 10 H. pullorum strains the 16S rRNAs were sequenced. The developed PCR is a helpful diagnostic tool to detect and differentiate Campylobacter, Arcobacter and Helicobacter isolated from poultry and poultry products.     

Survival of Escherichia coli,Enterococci and Campylobacter jejuni in Canada Goose Faeces on Pasture

Freshly excreted Canada goose faeces pose a public health risk as they contain pathogenic microorganisms. Accordingly, a study was carried out on the growth and survival of resident indicator bacteria (enterococci and Escherichia coli) and inoculated Campylobacter jejuni in freshly excreted faeces over summer and winter. Canada goose faeces were collected, mixed thoroughly and inoculated with 10⁸ g^?1C. jejuni. The faeces were mixed again before making the Canada goose dropping. The simulated goose droppings (N = 70) were placed on pasture, and the concentrations of E. coli, enterococci and the pathogen, C. jejuni, were monitored. In summer only, the molecular marker of E. coli LacZ and the avian‐associated bacteria E2 was also monitored. Results of the survival study indicated that significant growth of enterococci and E. coli occurred in summer, before concentrations decreased to less than 15% of the original concentration (day 77) for enterococci and 0.01% for E. coli. LacZ followed a similar pattern to E. coli, while the E2 marker dropped to below 0.1% of the original concentration within 4 days. In winter, enterococci grew slightly, while no growth of E. coli occurred. In both summer and winter, C. jejuni was rapidly inactivated. This research highlights the ability of bacterial indicators to replicate and survive in the environment when harboured by avian faeces, and the limited risk aged Canada goose faeces pose as an environmental source of Campylobacter spp.     

Isolation of Campylobacter spp. from Client‐Owned Dogs and Cats,and Retail Raw Meat Pet Food in the Manawatu,New Zealand

Campylobacter causes acute gastroenteritis in people worldwide and is frequently isolated from food, animals and the environment. The disease is predominately food‐borne but many routes of transmission and sources of infection have been described, including contact with pets. The prevalence of Campylobacter spp. in dogs and cats varies widely, and data on New Zealand pets are limited. This study aimed to investigate the prevalence of Campylobacter spp. in dogs, cats and retail raw meat pet food products in New Zealand and to characterize Campylobacter jejuni isolates using multilocus sequence typing (MLST). Ninety dogs and 110 cats examined at the Massey University Veterinary Teaching Hospital for elective procedures, and fifty locally purchased retail raw meat pet diets were sampled. Two culture protocols combining Bolton broth enrichment and mCCDA and CAT agars in a microaerobic atmosphere at 42°C and 37°C with species identification using PCR were performed. The prevalence of Campylobacter spp., C. jejuni, Campylobacter upsaliensis and Campylobacter helveticus was 36%, 13%, 23% and 1% in dogs and 16%, 5%, 5% and 7% in cats, respectively. One dog had Campylobacter lari confirmed, and three dogs and one cat had multiple Campylobacter spp. detected. Significantly more animals tested positive using CAT than mCCDA agar (P < 0.001). Being neutered, vaccinated for Bordetella bronchiseptica, fed dry diets and brought in for neutering were protective factors for dogs, whereas attendance for dental treatment was a risk factor for cats. Campylobacter spp. were isolated from 28%, C. jejuni 22%, C. lari 6% and Campylobacter coli 6% of food samples. Six isolates positive by Campylobacter genus PCR were identified as Arcobacter butzleri. Poultry meat was more likely to be positive than non‐poultry meat (P = 0.006). Of the 13 C. jejuni pet isolates with full MLST profiles, eight were of different sequence types (ST) and all nine food isolates were of different STs.     

Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.     

Molecular and Statistical Analysis of Campylobacter spp. and Antimicrobial‐Resistant Campylobacter Carriage in Wildlife and Livestock from Ontario Farms

The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial‐resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross‐sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial‐resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter‐specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C.  coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between livestock and wildlife samples, and the lack of similarity in molecular subtypes and AMR patterns, we concluded that the sharing of Campylobacter species between livestock and mammalian wildlife was uncommon.     

Molecular epidemiology of Campylobacter isolates from broiler slaughterhouses in Tripoli,North of Lebanon

ABSTRACT

1. The real burden of Campylobacter spp. in Lebanon is still unknown. The aims of this study were to unravel the epidemiology of Campylobacter spp. in broilers at slaughterhouses in Tripoli, North of Lebanon and to characterise their antibiotic resistance profiles.

2. From May to November 2015, sampling was performed through five repeated surveys from 15 slaughterhouses that sold chicken directly to Lebanese customers. Isolates were subjected to pulsed field gel electrophoresis (PFGE) and flaA-restriction fragment length polymorphism (flaA-RFLP).

3. All investigated slaughterhouses were found to be positive for Campylobacter spp. Campylobacter coli was the predominant species (38 isolates) followed by C. jejuni (eight isolates). A noticeable level of resistance was detected among isolates against ciprofloxacin (97% of C. coli and 87.5% of C. jejuni), amoxicillin (89% of C. coli and 75% of C. jejuni), gentamicin (79% of C. coli and 50% of C. jejuni), and co-amoxiclav (24% of C. coli and 25% of C. jejuni). Erythromycin and ertapenem resistance were observed only in C. coli with the following percentages 74% and 13% respectively, but not in C. jejuni. PFGE and flaA-RFLP using DdeI as restriction enzyme divided the strains into 27 and 25 types respectively.

4. The high observed genetic diversity of Campylobacter spp. revealed the complexity of the spread of this genus in broilers. This study highlighted the pressing need to monitor antibiotic resistance and to ensure food safety from ‘farm to fork’ in Lebanon.     

The occurrence and significance ofCampylobacter jejuni in man and animals

Campylobacter jejuni, which is now recognized as a discrete species, is a gram negative, microaerophilic, thermophilic, nalidixic acid sensitive, hippurate positive pathogen requiring special selective media for propogation. The organism is widely distributed in avian species, experimental and companion animals and in humans. Mammalian campylobacteriosis is characterized by an enterocolitis of variable severity. The prevalence of the condition is relatively high in young individuals, in underdeveloped countries and in subjects with diarrhea. Food animals, especially poultry, are reservoirs of the organism and infection occurs following consumption of untreated surface water, unpasteurized milk, incompletely cooked meat or other contaminated food products. Close contact with infected immature companion animals is a significant cause of campylobacteriosis in children and direct intrafamilial transmission and occupational infection have been documented.Campylobacteriosis attributable toC. jejuni is a condition of emerging significance which arises principally from deficiencies in hygiene inherent in the environment and in the food chain which extends from domestic animals to the consumer.     

Prevalence and risk factors of Campylobacter infection in broiler flocks from southern Spain

An extensive epidemiological study was performed to determine the prevalence and risk factors of Campylobacter infection in broiler farms in Andalusia (southern Spain). A total of 2221 cloacal swabs and 747 environmental swabs from 291 broiler flocks were screened between April 2010 and May 2012. The prevalence of Campylobacter in individual animals was 38.1%, and the flock prevalence was 62.9%. Flocks were predominantly infected by C. jejuni and C. coli but were also infected by untyped Campylobacter spp., and mixed-species infection could be found. Risk factors for Campylobacter infection were assessed from direct interview of the farmers. The number of positive samples by flock was modelled assuming a binomial distribution. Analysis indicated five factors associated with increased intra-flock prevalence: presence of dogs or cats on the farm, older age of the broiler flock, the application of thinning of flocks, the presence of windows with canvas blinds, and the presence of rodents in the poultry house. Two factors were associated with decreased intra-flock prevalence: the treatment of drinking water and having an entrance room for access into the poultry house. This is the first study performed on broilers farms from Spain reporting the risk factors of Campylobacter infection and is the largest study on the prevalence of Campylobacter infection.