Occurrence and genotypes of Campylobacter in broiler flocks, other farm animals, and the environment during several rearing periods on selected poultry farms

On 15 Swiss poultry farms, broiler flocks, other farm animals, and the environment were examined during consecutive rearing periods to investigate the occurrence and genetic diversity of Campylobacter. Of the 5154 collected samples, 311 (6%) from 14 farms were Campylobacter positive by culture. Amongst the positive samples, 228 tested positive for Campylobacter jejuni and 92 for Campylobacter coli. Positive samples originated from broilers, the broiler houses, cattle, pigs, bantams, laying hens, a horse, and a mouse. Feed, litter, flies, and the supply air to the broiler house tested negative. By flagellin gene typing (fla-RFLP) and pulsed-field gel electrophoresis (PFGE), 917 Campylobacter isolates were genotyped. Additionally, amplified fragment length polymorphism (AFLP) analysis was performed on 15 assorted strains. On eight farms, matching genotypes were isolated from broiler flocks and other farm animals: Certain genotypes from cattle (farms H, K, L, and M), pigs (farms D and P), or laying hens (farm L) were subsequently found in the broiler flocks, whereas other genotypes initially present in the broiler flocks turned up in cattle (farms A, D, and O). These results emphasize the importance of other farm animals on poultry farms for broiler flock colonization. Indications of persistent contamination of the broiler house were evident on four farms (C, D, I, and L) where matching genotypes were detected in consecutive broiler flocks, but not concurrently in other samples. By fla-RFLP, PFGE, and confirmed by AFLP, some genotypes proofed to be identical across different farms.     

Prevalence and characterization of Salmonella infantis isolates originating from different points of the broiler chicken-human food chain in Hungary

During the 10-month study period Salmonella contamination of broiler houses and the flocks reared in three farms (A, B and C), the slaughter houses where the flocks were slaughtered, as well as the carcass and retail raw meat products originating from them was investigated. In the broiler farm A five consecutive flocks, in the B and C farms one flock was sampled. Environmental samples were taken prior to the introductions. Environmental, drinking water, feed and faecal samples were collected regularly using standard methods. Before and during processing of the flocks, environmental and carcass samples were taken at the abattoirs. Salmonella contamination of the carcass, retail meat, as well as stool samples of farm and abattoir workers and from human illnesses registered in the same period and region were also examined. Isolation, sero-, phage- and antibiotic resistance typing, class 1 integron and plasmid profiling of the strains were performed; their genetic relationship was assessed by PFGE. Although the broiler house and the faecal samples of the 5 flocks of the farm A were negative for Salmonella, S. infantis was isolated from 20-100% of the abattoir carcass samples. The retail raw meat samples were 0-100% S. infantis positive. The environmental samples of farm B were Salmonella negative, but the examined flock was contaminated: S. infantis was identified from 43% of the faecal samples. This serotype was identified in 100% of the carcass and retail raw meat samples. From environmental samples taken before the arrival of the 1-day-old chicks in the broiler house C, S. infantis was cultured. S. infantis prevalence in the faecal samples was 35% and all the carcass and retail raw meat samples were S. infantis contaminated. Altogether 164 S. infantis strains were isolated out of which 145 were further characterized. The vast majority (142/145) of the strains belonged to phage types 217 and 213. All but one were characterized by the nalidixic acid-streptomycin-sulphonamide-tetracycline resistances, had an 885 bp class 1 integron and a large plasmid of >168 kb in size. The strains showed >/=88.7% genetic similarity. The results obtained shows that the same multi-drug resistant S. infantis clone was spread from the examined broiler farms contaminating the slaughter and the retail meat and appeared in the human illnesses of the examined region that was earlier detected as the dominant clone characteristic of the broiler and human population of the whole country.     

Prevalence of Campylobacter spp. in poultry and poultry meat in Germany

Of 509 samples from poultry flocks, 209 isolates (41.1%) were Campylobacter positive. The number of positive cases in broiler carcasses was 45.9%. Of 52 pheasants investigated, 25.9% were Campylobacter positive. Campylobacter jejuni was isolated from 86 (42.0%) poultry flock samples, 47 (43%) broiler samples and 15 (28%) wild pheasant samples. C. coli was found at a rate of 1.2% in poultry flocks, 13% in broilers and 21% in pheasants.     

Genotyping of Campylobacter coli and C. jejuni from retail chicken meat and humans with campylobacteriosis in Slovenia and Bosnia and Herzegovina

Thermotolerant Campylobacter jejuni and C. coli are one of the major causes of bacterial foodborne enteric infection. Consuming and/or handling poultry meat is the most consistent risk factor, linked to the high prevalence of campylobacters in retail poultry meat. The aim of the present study was to ascertain the genetic diversity and/or possible specificity of thermotolerant Campylobacter isolates according to species (C. coli, C. jejuni), isolation source (retail chicken meat and human clinical samples) and geographic origin (Goriska in Slovenia and Zenica-Doboj Canton in Bosnia and Herzegovina (BIH)). With the pulsed-field gel electrophoresis (PFGE) after SmaI macrorestriction we distinguished 80 PFGE types among 118 strains and CfoI restriction fragment length polymorphism of the amplified flagellin gene (fla-RFLP) gave 12 fla-RFLP types. Beside the higher discriminatory power and strain typeability, PFGE discriminated the C. jejuni and C. coli groups of isolates. A high proportion of C. coli strains was isolated, especially from poultry samples. Identical or very similar PFGE types among the isolates from animal, food and human samples indicate the transmission of C. jejuni and C. coli from the chickens on the farm to the retail chicken meat, as well as possible cross-contamination of retail meat and transmission to humans. However, the identity of the isolates from non-related samples but with identical PFGE and fla-RFLP types should be confirmed with additional typing. Reliable tracing of the source of Campylobacter strains by molecular typing of the chicken meat isolates is therefore very difficult. The reasons include contamination of meat samples with multiple strains, possible cross-contamination and extreme heterogeneity of the isolates (mainly for C. jejuni) on one side and a limited power of the genotyping methods used to distinguish non-related strains on the other side (mainly for C. coli).     

Genotyping of broiler-originated Campylobacter jejuni and Campylobacter coli isolates using fla typing and random amplified polymorphic DNA methods

Liver and intestine samples taken from 200 broilers at 20 flocks were inoculated onto Preston Enrichment broth and agar for selective isolation of Campylobacter jejuni and Campylobacter coli. The isolates were identified by both conventional and polymerase chain reaction (PCR) methods. Campylobacter spp. were identified in 102 of 400 samples (200 liver and 200 intestine), 57 (14.25%) of which were identified as C. jejuni and 45 (11.25%) as C. coli. PCR-restriction fragment length polymorphism (RFLP) of the flagellin gene (flaA) and random amplified polymorphic DNA (RAPD) typing were used to describe the heterogeneity among amplified DNA products of C. jejuni and C. coli isolates. Flagellin gene analysis by RFLP of the isolates produced seven different band profiles. On the other hand, five distinct band profiles were obtained in the examination of the isolates with RAPD assay using a random primer (OPA-11). The results of this study demonstrated that a relatively low heterogeneity existed among C. jejuni and C. coli strains isolated from the commercial broiler flocks in eastern Turkey. In the comparison of both typing methods, fla typing provided more discrimination than the RAPD assay used.     

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

In this study Campylobacter jejuni isolates were recovered from birds, carcasses and carcass portions from two broiler chicken flocks and from equipment used for carcass and meat processing along the production chain from farms to retail stores. Isolates were subjected to pulsed-field gel electrophoresis (PFGE) using SmaI and KpnI restriction enzymes and their antimicrobial susceptibilities were determined. C. jejuni was recovered from product and equipment used with both flocks at each point in the production chain. The prevalence of C. jejuni in poultry products at retail stores was 58.97% (flock 1) and 69.23% (flock 2). SmaI divided 122 C. jejuni strains from flock 1 and 106 from flock 2 into 17 and 13 PFGE types, respectively. PFGE types H and F were present at all steps along the chain, from farms to retail products. Similarly, for both flocks PFGE type D was detected in crates, slaughterhouse and retail stores. Moreover, the PFGE types were highly diverse at the processing and retail steps. Most PFGE types were resistant to ciprofloxacin (95.45%) and tetracycline (81.82%); and multidrug resistant PFGE types were found in the final products. Our study showed that there were several points of cross-contamination of product along the chain, and a high diversity of PFGE types with antimicrobial resistance to ciprofloxacin and tetracycline in the retail products.     

Molecular epidemiology and antimicrobial susceptibility of Campylobacter jejuni and Campylobacter coli isolates of poultry, swine, and cattle origin collected from slaughterhouses in Hungary

Campylobacter spp. are the most common cause of bacterial enteritis in Hungary, and the aim of this study was to identify the distribution, genotypes, and antimicrobial susceptibility of Campylobacter species in the most important food-producing animals at the time of slaughter during 2008 and 2009. Of 1,110 samples, 266 were identified as Campylobacter coli (23.9%) and 143 as C. jejuni (12.9%) by real-time PCR. Resistance to enrofloxacin-ciprofloxacin and nalidixic acid was significant, especially in C. jejuni (73.3%) and C. coli (77.2%) from broilers. Higher erythromycin (P = 0.043) and tetracycline (P = 1.865e-14) resistance rates were found among C. coli isolates (9.7 and 74.1%, respectively) than among C. jejuni isolates (3.1 and 36.6%, respectively). A total of 47 fla short variable region sequences were identified among 73 selected C. coli and C. jejuni isolates, with 35 fla types detected only once. At the nucleotide level, fla types A66 and A21 were the most common. Using the pulsed-field gel electrophoresis method, 66% of strains exhibited unique profiles after Sma I digestion. Forty-two isolates assigned to 18 Sma I clusters were further typed by Kpn I, and of these, 24 were assigned to 10 Kpn I clusters. For isolates in five Kpn I clusters, epidemiological links were observed. Stable C. jejuni and C. coli clones were detected, indicating that further studies involving broiler and human isolates need to be conducted to elucidate the importance of these stable clones in human infections.     

Reduction in flock prevalence of Campylobacter spp. in broilers in Norway after implementation of an action plan

An action plan against thermophilic Campylobacter spp. in Norwegian broilers was implemented in May 2001. The action plan consists of three parts: a surveillance program including all Norwegian broiler flocks slaughtered before 50 days of age, a follow-up advisory service on farms delivering flocks positive for Campylobacter spp., and surveys of broiler meat products at the retail level. This article presents results covering the inclusive 3-year period between 2002 and 2004. During this period, a total of 10,803 flocks from 562 broiler farms were tested; altogether, 521 (4.8%) of the flocks were identified as positive for Campylobacter spp., primarily Campylobacter jejuni. The positive flocks originated from 257 (45.7%) of the farms. During the period 2002 to 2004, there was a large and steady reduction in flock prevalence, from 6.3% in 2002 to 3.3% in 2004. Also, the proportion of farms producing flocks positive for Campylobacter spp. each year reduced substantially, from 28.4% in 2002 to 17.8% in 2004. The proportion of flocks positive for Campylobacter spp. varied considerably with season and region. The action plan is a successful collaboration between academia, regulatory agencies, and the poultry industry that has resulted in a significant reduction in the number of broiler carcasses positive for Campylobacter spp. on the market. The temporal associations between implementation of the control program and the drop in the number of infected chickens and contaminated carcasses indicate that this collaborative action plan has been instrumental in achieving the goals of enhancing food safety.     

Prevalence and comparison of genetic profiles of Campylobacter strains isolated from poultry and sporadic cases of campylobacteriosis in humans.

Between July 1998 and June 1999, 93 lots of broiler chickens distributed on 57 farms were sampled in two abattoirs of the province of Quebec (Canada). A total of 2,325 samples of cecal material were analyzed to determine the prevalence of campylobacters. Biotyping and pulsed-field gel electrophoresis (PFGE) were done on 20% of the Campylobacter isolates to study the distribution within poultry production. Macrorestriction profiles were compared with profiles of 24 Campylobacter strains isolated from sporadic cases of human diarrheic patients in order to evaluate genetic relationships. Approximately 40% of the broiler chickens in 60% of the lots and 67% of the farms were colonized. Biotypes I and II of Campylobacter jejuni were the most prevalent biotypes in poultry and human isolates. The PFGE dendograms revealed a high genetic diversity among poultry isolates, with 49 different genotypes from the 56 positive lots. More than 75% of these lots were colonized by a unique genotype. All positive lots raised simultaneously on the same farm had common genotype(s). Different genotypes were isolated from lots raised at different grow-out periods on a farm. In some cases, identical genotypes were found at different grow-out periods on a farm and also from different farms. Macrorestriction profiles showed that approximately 20% of human Campylobacter isolates were genetically related to genotypes found in poultry. This genetic relationship and the high prevalence of C. jejuni biotypes I and II in poultry indicated that Campylobacter in broiler production of the province of Quebec could be a potential source of hazard for public health.     

Antimicrobial resistance and molecular subtyping of Campylobacter jejuni and Campylobacter coli from retail meats

Campylobacter isolates (n = 297; 202 C. jejuni and 95 C. coli isolates) recovered from 2,513 retail meat samples (chicken breasts, ground turkey, ground beef, and pork chops) were examined for antimicrobial susceptibility. The isolates were further analyzed for genetic relatedness by pulsed-field gel electrophoresis (PFGE) using SmaI and KpnI restriction enzymes, and a subset of isolates (n = 174) were subtyped by multilocus sequence typing (MLST). The resistance most frequently observed was that to doxycycline (27.6%), followed by ciprofloxacin (13.8%) and erythromycin (6.4%). All isolates were susceptible to gentamicin and meropenem. C. coli showed higher resistance to doxycycline than did C. jejuni (42.1 versus 20.8%) and lower resistance to ciprofloxacin than did C. jejuni (10.5 versus 15.3%). Erythromycin resistance was only observed in C. coli. PFGE using SmaI plus KpnI digestion generated 168 clusters from 297 isolates: 115 from C. jejuni and 53 from C. coli. MLST revealed 44 sequence types (STs) under 10 clonal complexes from 120 C. jejuni and 27 STs under two clonal complexes from 54 C. coli. There was a positive association between PFGE and STs; however, PFGE showed greater discriminatory power than MLST. Subtyping data did not correlate with antimicrobial resistance phenotypes.     

A survey of food-borne pathogens in free-range poultry farms

A survey of the occurrence of Campylobacter, Salmonella, Listeria and Shiga toxin-producing E. coli was performed on 60 flocks of free-range chicken from 34 farms in the Basque Country (Northern Spain). Campylobacter was the most prevalent of the four pathogens, isolated in 70.6% of the farms, followed by L. monocytogenes (26.5%), and Salmonella (2.9%). No E. coli O157 or other STEC were isolated. In total 48 flocks from 26 farms were positive for at least one pathogen: 31 of them for a single pathogen (64.6%), and 17 for more than one species (35.4%). C. coli was more prevalent than C. jejuni (15 vs. 13 farms), and both species of Campylobacter were found in 3 farms. L. monocytogenes isolates were identified as serotype 4b complex, and the only Salmonella isolated was serovar Enteritidis. flaA PCR-RFLP performed on 91 Campylobacter isolates (36 C. jejuni and 55 C. coli) yielded 26 patterns, with higher diversity among the C. jejuni isolates. More than one pattern was found in 11 farms, and in 8 of them several patterns were found within the same flock. The findings of the present study suggest that the free-range rearing conditions described herein might have an advantageous effect on diminishing Salmonella but not on Campylobacter or L. monocytogenes flock contamination.     

Tracing flock-related Campylobacter clones from broiler farms through slaughter to retail products by pulsed-field gel electrophoresis

A total of 237 Campylobacter isolates from broiler flocks at farm (45 isolates) and slaughter (192 isolates) were typed by pulsed-field gel electrophoresis (PFGE) for epidemiological tracing studies. For PFGE, a modification of the Campynet method was used, which was standardized in a European Union project. The goal of the study was to trace flock-related Campylobacter clones through the whole production chain, from farm through slaughter to retail products, to investigate the introduction of thermophilic Campylobacter spp. on incoming contaminated carcasses during processing to the final products. The results of this study showed that identical clones of this pathogen, which had previously been found within the flocks during primary production, were also detected at individual stages of processing, including final products, which were packed and ready for sale. Most of the detected clones dominated during primary production and at slaughter. This study found PFGE to be suitable for examining epidemiological field data in the same region and time contexts. The discriminatory power of SmaI restriction enzyme digestion was sufficient. Relationships of the isolated Campylobacter strains could be confirmed by use of a second restriction enzyme, KpnI.     

Prevalence and characterization of Campylobacter jejuni isolated from pasture flock poultry

The growing interest in organic and natural foods warrants a greater need for information on the food safety of these products. In this study, samples were taken from 2 pasture flock farms (N = 178; feed, water, drag swabs, and insect traps), pasture flock retail carcasses (N = 48) and 1 pasture flock processing facility (N = 16) over a period of 8 mo. A total of 105 Campylobacter isolates were obtained from 53 (30%), 36 (75%), and 16 (100%) samples from the farms, retail carcasses, and processing facility, respectively. Of the 105 isolates collected, 65 were C. jejuni, 31 were C. coli, and 9 were other Campylobacter spp. Using PCR, the C. jejuni isolates were further analyzed for virulence genes involved in colonization and survival (flaA, flaC, cadF, dnaJ, racR, cbrR), invasion (virB11, ciaB, pldA), protection against harsh conditions (sodB, htrA, clpA), toxin production (cdtA, cdtB, cdtC), siderophore transport (ceuE), and ganglioside mimicry (wlaN). In addition, the short variable region of the flaA locus (flaA SVR) was sequenced to determine the genetic diversity of the C. jejuni isolates. The flaA SVR diversity indices increased along the farm to carcass continuum. PCR-based analysis indicated a low prevalence of 5 genes involved in colonization (dnaJ, ciaB, pldA, racR, virB11). The results of this survey indicate that the prevalence of Campylobacter on organic retail carcasses is similar to prevalence reports of Campylobacter on conventional retail carcasses. However, the genetic diversity of the flaA SVR genotypes increased along the farm to carcass continuum that contrasted with conventional poultry studies. PRACTICAL APPLICATION: Campylobacter jejuni is a leading cause of foodborne illness with poultry and poultry products being leading sources of infection. Free-range and pasture flock chickens are becoming more popular; however, there is an inherent biosecurity risk that can increase the prevalence of foodborne pathogens in these flocks. This study aimed to determine sources and characterize C. jejuni isolated from pasture flocks.     

Antimicrobial resistance in Campylobacter jejuni from broilers and broiler house environments in Norway

Antimicrobial susceptibility in Campylobacter jejuni collected from the environment outside four broiler houses (n = 63) and from the environment inside these broiler houses (including broiler droppings) (n = 36) from May to September 2004 was studied and compared with isolates from Norwegian broilers analyzed within the frame of the Norwegian monitoring program of antimicrobial resistance in feed, food, and animals (NORM-VET) in 2004 (n = 75). The MICs of oxytetracycline, ampicillin, erythromycin, gentamicin, enrofloxacin, and nalidixic acid were obtained by the broth microdilution method VetMIC. The present study, which to our knowledge is the first Norwegian study on the occurrence of antimicrobial resistance in Campylobacter spp. from the environment of broiler houses, revealed a very low occurrence of antimicrobial resistance in C. jejuni from the broilers and broiler house environments studied. All isolates originating from the four broiler houses studied were susceptible to all the antimicrobial agents tested, except for one isolate from the outdoor environment (courtyard soil), which was resistant to oxytetracycline (MIC, 8 mg/liter). For the isolates from broilers (NORM-VET), low prevalences of resistance to oxytetracycline (1.3%) and ampicillin (4%) were observed. No quinolone resistance was observed. The results for the broiler isolates are in agreement with the earlier findings of a very low prevalence of resistance in Campylobacter from broilers in Norway, which reflects the low usage of antimicrobials in Norwegian broiler production. Furthermore, the present data are in accordance with antimicrobial susceptibility data for C. jejuni from domestically acquired human cases.     

PFGE genotyping and antimicrobial susceptibility of Campylobacter in retail poultry meat in Estonia

In the present study, the Campylobacter isolates from retail poultry meat in Estonia were sero- and genotyped, and the antimicrobial susceptibility was determined. Forty-eight chicken (36 Estonian, 12 imported) and 22 turkey (imported) Campylobacter isolates from 580 raw broiler chicken (396 Estonian, 184 imported) and 30 turkey (imported) meat samples were studied. Of the isolates, 64 were C. jejuni, 4 C. coli, and 2 Campylobacter spp. Penner serotyping of 54 C. jejuni isolates revealed 11 different serotypes, and 22% of the isolates were nontypeable by the commercial antisera. The most common serotypes O:1,44; O:21, and O:55 accounted for 28%, 13%, and 13% of the isolates, respectively. Differences in serotype distribution were seen for chicken and turkey isolates. Genotypic characterization of all Campylobacter isolates (n=70) was performed by pulsed-field gel electrophoresis (PFGE). SmaI and KpnI yielded 29 and 34 PFGE types, respectively, revealing high diversity among isolates. The serotype distribution did not show an association with the origin of the sample, but the majority of the isolates sharing a similar PFGE genotype originated from one country. High levels of resistance to ciprofloxacin (66%), nalidixic acid (66%), tetracycline (44%), ampicillin (34%), and erythromycin (14%) were detected among the 70 Campylobacter isolates. The simultaneous resistance to two or three antimicrobial agents occurred in 60% of the isolates. The Campylobacter isolates from turkey meat had higher resistance to ampicillin, ciprofloxacin, nalidixic acid, and tetracycline than those from chicken meat. None of the chicken isolates were resistant to gentamicin, and no turkey isolates to erythromycin or gentamicin.     

Prevalence of Listeria monocytogenes in broilers at the abattoir, processing plant, and retail level.

The environment and products from two broiler abattoirs and processing plants and raw broiler pieces at the retail level were sampled for Listeria monocytogenes in order to evaluate the contamination level of the broiler carcasses and products. Sampling started in the slaughtering process and finished with raw broiler meat or ready-to-eat cooked product. Sampling sites positive for L. monocytogenes at the broiler abattoir were the air chiller, the skin-removing machine, and the conveyor belt leading to the packaging area. The L monocytogenes contamination rate varied from 1 to 19% between the two plants studied. Furthermore, 62% (38 of 61) of the raw broiler pieces, bought from retail stores, were positive for L. monocytogenes. Altogether, 136 L. monocytogenes isolates were obtained for serotyping and pulsed-field gel electrophoresis (PFGE) characterization performed with two rare-cutting enzymes (ApaI and AscI). Altogether three serotypes (1/2a, 1/2c, and 4b) and 14 different PFGE types were obtained using information provided from both ApaI and AscI patterns for discrimination basis. The two broiler abattoirs studied did not share the same PFGE types. However, the same PFGE types found in the raw broiler pieces at the retail level were also found in the broiler abattoirs where the broilers had been slaughtered.     

Multidrug resistant clones of Salmonella Infantis of broiler origin in Europe

The aim of the study was to investigate the potential spread of the previously described multidrug-resistant (MDR) Hungarian clone of Salmonella Infantis of broiler origin in Europe. Therefore, 76 strains isolated between 2004 and 2009 from raw meat and fecal samples of broiler origin in nine European countries - including Hungary - were examined by phage typing, antimicrobial resistance typing, pulsed-field gel electrophoresis (PFGE) profile and plasmid analysis. The strains could be divided into two large PFGE clusters with 92% similarity. Cluster A (n=39) contained 15 German, seven Italian, five British, five Polish isolates, one Austrian and one Hungarian isolate. Five Hungarian isolates that were isolated prior to the appearance of the MDR clone also belonged to this cluster. Strains of this cluster comprised seven pulsotypes and 14 different phage types and were mostly susceptible to the 12 antimicrobials tested. Cluster B (n=33) contained all but one of the recent Austrian (n=14) and of Hungarian (n=9), six Polish, one Italian and one German as well as the single Turkish and the Romanian strains, representing five pulsotypes. The strains of this cluster were mostly PT213, with MDR (nalidixic acid-streptomycin-sulphomamide-tetracycline), and the carriage of the same class 1 integron located on a large plasmid (>168kb) characteristic of the current predominant Hungarian clone. The results suggest that two large related clusters (A and B) of S. Infantis isolates can be found in various European countries, of which Austrian and Polish MDR clones of cluster B are identical with, or closely related to, the dominant Hungarian clone. The emergence of a few dominant MDR S. Infantis clones in broilers reported here, raises the possibility that further dissemination of such clones in broilers and in broiler meat may occur and represent a potential threat to public health in Europe.     

A multiplex polymerase chain reaction for the differentiation of Campylobacter jejuni and Campylobacter coli from a swine processing facility and characterization of isolates by pulsed-field gel electrophoresis and antibiotic resistance profiles

A multiplex polymerase chain reaction (PCR) was developed for the detection and speciation of 60 Campylobacter strains isolated from porcine rectal swabs and from different areas in a pork processing plant. The PCR assay was based on primers specific for the cadF gene of pathogenic Campylobacter species, a specific but undefined gene of Campylobacter jejuni, and the ceuE gene of Campylobacter coli. Further characterization of these isolates was established by pulsed-field gel electrophoresis (PFGE) analyses with the restriction endonuclease SmaI. In addition to molecular discrimination, the antibiotic resistance profiles of the isolates were examined by the Kirby Bauer disc diffusion method with 22 antibiotics. Differentiation of isolates by multiplex PCR identified 86.9% (52 of 60) as C. coli and 13.1% (8 of 60) as C. jejuni. Using the Molecular Analyst software, 60 PFGE types were identified. The percentages of relatedness among C. jejuni strains with PFGE ranged from 25 to 86%, while those among C. coli strains ranged from 34 to 99%. Among the 60 PFGE types, each of 12 C. coli isolates showed > or =90% similarity to one other isolate. The antibiotic resistance profiles of all 60 isolates were distinct. Analyses of antibiotic resistance profiles showed that all isolates were resistant to five or more antibiotics. Twenty-five percent (2 of 8) of C. jejuni isolates and 15% (8 of 52) of C. coli isolates were resistant to at least one of the three fluoroquinolones tested, antibiotics that are commonly used in the treatment of human Campylobacter infections. Three percent (2 of 60) of Campylobacter isolates examined were resistant to all three fluoroquinolones. On the basis of the PFGE and antibiotic resistance profiles, each of the 60 isolates was distinct, suggesting that C. jejuni and C. coli strains originating from diverse sources were present in porcine samples and in the pork processing plant.     

Contamination of meat with Campylobacter jejuni in Saitama, Japan.

To determine the source of food contamination with Campylobacter jejuni, we investigated retail meat, a chicken processing plant and a broiler farm. C. jejuni was found in domestic retailed poultry (45.8%) and imported poultry (3.7%), but not in beef or pork. In the poultry processing plant, there is significant contamination with C. jejuni in chicken carcasses, equipment and workers' hands. This contamination increases during the defeathering and evisceration processes. RAPD analysis shows that contamination with C. jejuni is of intestinal origin. In a broiler farm, C. jejuni was first isolated from a faecal sample of broiler chicken after the 20th day of age. Two weeks later, all birds in this farm became C. jejuni positive. RAPD analysis indicated that C. jejuni spread rapidly from one broiler flock to the other flocks on the farm.     

Use of MIDI-fatty acid methyl ester analysis to monitor the transmission of Campylobacter during commercial poultry processing

The presence of Campylobacter spp. on broiler carcasses and in scald water taken from a commercial poultry processing facility was monitored on a monthly basis from January through June. Campylobacter agar, Blaser, was used to enumerate Campylobacter in water samples from a multiple-tank scalder; on prescalded, picked, eviscerated, and chilled carcasses; and on processed carcasses stored at 4 degrees C for 7 or 14 days. The MIDI Sherlock microbial identification system was used to identify Campylobacter-like isolates based on the fatty acid methyl ester profile of the bacteria. The dendrogram program of the Sherlock microbial identification system was used to compare the fatty acid methyl ester profiles of the bacteria and determine the degree of relatedness between the isolates. Findings indicated that no Campylobacter were recovered from carcasses or scald tank water samples collected in January or February, but the pathogen was recovered from samples collected in March, April, May, and June. Processing generally produced a significant (P < 0.05) decrease in the number of Campylobacter recovered from broiler carcasses, and the number of Campylobacter recovered from refrigerated carcasses generally decreased during storage. Significantly (P < 0.05) fewer Campylobacter were recovered from the final tank of the multiple-tank scald system than from the first tank. MIDI similarity index values ranged from 0.104 to 0.928 based on MIDI-fatty acid methyl ester analysis of Campylobacterjejuni and Campylobacter coli isolates. Dendrograms of the fatty acid methyl ester profile of the isolates indicated that poultry flocks may introduce several strains of C. jejuni and C. coli into processing plants. Different populations of the pathogen may be carried into the processing plant by successive broiler flocks, and the same Campylobacter strain may be recovered from different poultry processing operations. However, Campylobacter apparently is unable to colonize equipment in the processing facility and contaminate broilers from flocks processed at later dates in the facility.