Campylobacter-negative Flocks Research Articles

Deciphering the Association between Campylobacter Colonization and Microbiota Composition in the Intestine of Commercial Broilers.

Campylobacter is a major food safety concern and is transmitted mainly via poultry meat. We previously found that some commercial broiler farms consistently produced Campylobacter-negative flocks while others were consistently Campylobacter-positive for consecutive production cycles although the farms operated under similar management practices. We hypothesized that this difference in Campylobacter colonization might be associated with the gut microbiota composition. To address this, six commercial broiler farms were selected based on their Campylobacter status (three negative and three positive) to evaluate the microbiota differences between each farm category. For each farm on each production cycle (2-3 cycles), 40 ceca collected from five-week-old broilers were processed for microbiota analysis via 16S rRNA gene sequencing. Cecal microbiota species richness, phylogenetic diversity, community structure, and composition of Campylobacter-positive farms were noticeably different from those of Campylobacter-negative farms. Rikenella, Methanocorpusculum, Barnesiella, Parasutterella, and Helicobacter were significantly more abundant among Campylobacter-positive farms. In contrast, Ruminococcaceae, Streptococcus, Escherichia, Eggerthellaceae, Lactobacillus, Monoglobus, and Blausia were more abundant in Campylobacter-negative farms. Eggerthellaceae, Clostridia, Lachnospiraceae, Lactobacillus, Monoglobus, and Parabacteroides were significantly negatively correlated with Campylobacter abundance. These findings suggest that specific members of cecal microbiota may influence Campylobacter colonization in commercial broilers and may be further explored to control Campylobacter in poultry.

An investigation of broiler caecalCampylobactercounts at first and second thinning

The objectives of this study were the following: (i) to investigate if Campylobacter negative flocks at first thinning remain negative at second thinning; (ii) to determine if the caecal counts in birds infected during first thinning remain lower than in birds that were positive at first thinning; and (iii) to determine if reducing the time between first and second thinning to a maximum of 4 days would reduce both the incidence and prevalence of broiler caecal contamination. Twenty-two flocks were tested at first and second thinning using ISO methodologies. Of the 14 that had a 4-day duration between first and second thinning, nine flocks were Campylobacter negative at first thinning. By second thinning, all 14 flocks were positive and Campylobacter counts ranged from 5·5 to 6·6 log(10) CFU g(-1) regardless of the status at first thinning. The other eight flocks were all Campylobacter positive at first thinning with counts ranging from 0·8 to 6·1 log(10) CFU g(-1) which increased to 5·1 to 6·9 log(10) CFU g(-1) by second thinning (3-10 days). PCR speciation and MLST genotyping suggested the majority of isolates were Camp. jejuni belonging to STs 257, 814, 6763 and 6764. It was concluded that; thinning introduces Campylobacter into broiler flocks; caecal counts in birds at second thinning are similar, regardless of flock status at first thinning and reducing the time between first and second thinning to a maximum of 4 days is not an effective control strategy. This study informs Campylobacter control strategy at primary production, suggesting all post-first thinning broilers should be treated as 'high risk' regardless of Campylobacter status at first thinning or duration between first and second thinning.

Quantitative estimation of Campylobacter cross-contamination in carcasses and chicken products at an abattoir

This study aimed to investigate Campylobacter contamination in carcasses and chicken products derived from a Campylobacter-negative flock when the flock is slaughtered immediately after a Campylobacter-positive flock. The first 2 flocks slaughtered on 10 different dates were investigated at an abattoir. Eighteen of the 20 flocks tested were positive for Campylobacter. A Campylobacter-negative flock was slaughtered immediately after a Campylobacter-positive flock on only 1 of the 10 slaughter dates. In this case, Campylobacter was detected in the carcasses and chicken products originating from the Campylobacter-negative flock, and all the flaA genotypes of these isolates were identical to those present in the caecal contents, carcasses, and chicken products from the Campylobacter-positive flock. The Campylobacter concentrations in the products originating from the Campylobacter-negative flock were: close to the enumeration limit (1.7 log10 cfu/carcass) in the carcass samples; and below the enumeration limit (2.0 log10 cfu/g) in the liver samples. The mean Campylobacter concentrations in the carcasses and liver products originating from the 18 Campylobacter-positive flocks were 3.8 log10 cfu/carcass and 2.6 log10 cfu/g, respectively. While 91% (246/270) of chicken products originating from Campylobacter-positive flocks were positive for Campylobacter, chicken products originating from the remaining Campylobacter-negative flock were free from Campylobacter cross-contamination by slaughter prior to a Campylobacter-positive flock. These results prove that slaughtering Campylobacter-negative flocks does not introduce Campylobacter into the abattoirs and indicate that although carcasses and chicken products originating from the Campylobacter-negative flock were cross-contaminated with Campylobacter from the Campylobacter-positive flock slaughtered immediately before, the Campylobacter contamination levels were lower than those in carcasses and chicken products from Campylobacter-positive flocks. Based on these findings, the reduction of Campylobacter prevalence in broiler flocks should be taken as an effective control measure for preventing introduction of Campylobacter into abattoirs and consequently for reducing Campylobacter prevalence in chicken products in addition to the good hygienic practice at abattoirs and logistic slaughter.

Campylobacter Cross‐Contamination of Chicken Products at an Abattoir

Consumption of raw or undercooked poultry products contaminated with Campylobacter has been identified as a risk factor for human campylobacteriosis. We determined whether slaughtering of Campylobacter-positive flocks was associated with contamination of chicken products derived from Campylobacter-negative flocks slaughtered at the same abattoir. The presence of Campylobacter was investigated in 22 broiler farms 1 week prior to slaughter and in one abattoir on nine separate slaughter days. A total of 600 bulk packed chicken products were tested, with 198 (33.0%) of the products found to be Campylobacter positive. Of the 350 chicken products originating from Campylobacter-positive flocks, 180 (51.1%) were contaminated with the bacteria. In contrast, only 18 (7.2%) of 250 chicken products derived from Campylobacter-negative flocks were contaminated. In 14 of these 18 products, the Campylobacter isolates were identical to isolates obtained from the flock slaughtered immediately prior to the Campylobacter-negative flock. Notably, on 4/6 slaughter days, Campylobacter-negative flocks were slaughtered prior to the positive flocks, and Campylobacter was absent from all chicken products originating from the negative flocks. These results suggest that implementation of logistic slaughter (where Campylobacter-negative flocks are slaughter first) significantly decreases the prevalence of Campylobacter-positive chicken products.

Molecular tracking, through processing, of Campylobacter strains colonizing broiler flocks.

Many of the poultry flocks produced in the United Kingdom are colonized with Campylobacter, and the intensive nature of poultry processing usually results in contaminated carcasses. In this study, a previously reported molecular oligonucleotide probe method was used to track a specific flock-colonizing strain(s) on broiler carcasses during processing in two United Kingdom commercial poultry processing plants. Five Campylobacter-positive flocks were sampled at four points along the processing line, postbleed, postpluck, prechill, and postchill, and two Campylobacter-negative flocks processed immediately after positive flocks were sampled prechill. flaA was sequenced from Campylobacter strains isolated from these flocks, and strain-specific probes were synthesized. Skin and cecal samples were plated onto selective agar to give individual colonies, which were transferred onto membranes. These were then hybridized with the strain- and genus-specific probes. For all the 5 positive flocks, there was a significant reduction in campylobacters postbleed compared to postpluck but no subsequent fall on sampling pre- and postchill, and the strain(s) predominating on the carcasses throughout processing came from the flock being processed. This indicates that strains from the abattoir environment were not a significant cause of carcass contamination in flocks with well-established campylobacter colonization. However, negative flocks that were preceded by positive flocks were contaminated by strains that did not generally originate from the predominating strains recovered from the ceca of the previous positive flocks. This suggests that the abattoir environment has a significant role in the contamination of carcasses from negative but not fully colonized flocks.

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

The aim of this study was to assess the diversity of thermotolerant Campylobacter spp. isolated from turkey flocks at six rearing farms 1-2 weeks prior to slaughter (360 faecal swab samples) and from 11 different stages at the slaughterhouse (636 caecal, environmental, neck skin and meat samples). A total of 121 Campylobacter isolates were identified to species level using a multiplex PCR assay and were typed by pulsed-field gel electrophoresis (PFGE) and flaA-short variable region (SVR) sequencing. All Campylobacter isolates were identified as Campylobacter jejuni. PFGE analysis with KpnI restriction enzyme resulted in 11 PFGE types (I-XI) and flaA SVR typing yielded in nine flaA-SVR alleles. The Campylobacter-positive turkey flocks A, C and E were colonized by a limited number of Campylobacter clones at the farm and slaughter. The present study confirms the traceability of flock-specific strains (PFGE types I, V and IX; flaA types 21, 36 and 161) from the farm along the entire processing line to meat cuts. It seems that stress factors such as high temperature of the defeathering water (54-56 °C), drying of the carcass skin during air chilling (24 h at 2 °C), and oxygen in the air could not eliminate Campylobacter completely. Campylobacter-negative flocks became contaminated during processing by the same subtypes of Campylobacter introduced into the slaughter house by preceeding positive flocks even if they were slaughtered on subsequent days. Proper and efficient cleaning and disinfection of slaughter and processing premises are needed to avoid cross-contamination, especially in countries with a low prevalence of Campylobacter spp. The majority of flaA SVR alleles displayed a distinct association with a specific PFGE type. However, a linear relationship for all strains among both typing methods could not be established. To specify genetic relatedness of strains, a combination of different genotyping methods, is needed.

Review of the Carriage of Zoonotic Bacteria by Arthropods, with Special Reference toSalmonellain Mites, Flies and Litter Beetles

This systematic review considers the relationship between arthropods commonly found in and around livestock premises and zoonotic bacteria. The principal focus is upon insects and arachnids on poultry units, where houses, litter and manure provide good conditions for the growth, multiplication and protection of flies, beetles and mites, and where zoonotic pathogens such as Salmonella and Campylobacter are prevalent. Other members of the Enterobacteriaceae and the taxa Clostridium, Helicobacter, Erysipelas and Chlamydiaceae are also discussed. Salmonella is widely distributed in the flies of affected livestock units and is detectable to a lesser degree in beetles and mites. Persistent carriage appears to be common and there is some field and experimental evidence to support arthropod-mediated transmission between poultry flocks, particularly carry-over from one flock to the next. Campylobacter may readily be isolated from arthropods in contact with affected poultry flocks, although carriage is short-lived. There appears to be a role for flies, at least, in the breaching of biosecurity around Campylobacter-negative flocks. The carriage of other zoonotic bacteria by arthropods has been documented, but the duration and significance of such associations remain uncertain in the context of livestock production.

Evaluation of Logistic Processing To Reduce Cross-Contamination of Commercial Broiler Carcasses with Campylobacter spp.

Cross-contamination of broiler carcasses with Campylobacter is a large problem in food production. Here, we investigated whether the contamination of broilers carcasses from Campylobacter-negative flocks can be avoided by logistic scheduling during processing. For this purpose, fecal samples were collected from several commercial broiler flocks and enumerated for Campylobacter spp. Based on enumeration results, flocks were categorized as Campylobacter negative or Campylobacter positive. The schedule of processing included the testing of Campylobacter-positive flocks before or after the testing of Campylobacter-negative flocks. During processing, flocks were also sampled for Campylobacter spp. before and after chilling. Campylobacter strains were identified with multiplex PCR and analyzed for relatedness with pulsed-field gel electrophoresis. Our results show that Campylobacter-negative flocks were indeed contaminated with Campylobacter strains originating from previously processed Campylobacter-positive flocks. Campylobacter isolates collected from carcasses originating from different farms processed on the same day showed similar pulsed-field gel electrophoresis patterns, confirming cross-contamination. These findings suggest that a simple logistic processing schedule can preserve the Campylobacter-negative status of broiler carcasses and result in products with enhanced food safety.

Campylobacter spp. contamination of chicken carcasses during processing in relation to flock colonisation

The presence and numbers of campylobacters on chicken carcasses from 26 slaughter groups, originating from 22 single-house flocks and processed in four UK plants, were studied in relation to the level of flock colonisation determined by examining the caecal contents of at least ten birds per group. The prevalence of campylobacters on carcasses from five campylobacter-negative flocks processed just after other negative flocks was low (≤ 30%). Campylobacters were isolated from 90 to 100% of carcasses from three flocks which were partly colonised, with 5, 5 and 30% of caecal contents positive, and which were processed after fully colonised flocks. All carcasses from the remaining fully colonised flocks were contaminated with campylobacters, and they had significantly ( P < 0.001) higher numbers per carcass (average of 5.3 log 10 cfu; range: 1.3 to > 8.0 log 10 cfu) than carcasses originating from low prevalence flocks (average of 2.3 log 10 cfu; range: < 1.1 to 4.1 log 10 cfu). There was a reduction in the numbers of campylobacters on carcasses between plucking and chilling in eight of ten fully colonised flocks. In another eight flocks, a significant ( P < 0.001) decrease (0.8 log 10 cfu) in the number of campylobacters on carcasses from just before to after chilling was detected. Campylobacter spp. could be isolated from aerosols, particles and droplets in considerable numbers in the hanging-on, defeathering and evisceration areas but not in the chillers. This was the case even when campylobacters were not isolated from the target flock. Campylobacters on carcasses from two partly colonised flocks were either the same subtype, as determined by speciation, Multi-Locus Sequence Typing (MLST) and flaA Restricted Fragment Length Polymorphism (RFLP) typing, as those in the fully colonised flocks processed previously, although not necessarily the most prevalent ones; or were the same subtypes as those found in the caeca of the flock itself. The prevalences of the different campylobacter subtypes found on carcasses from two fully colonised flocks did not closely reflect those found in the caeca. MLST combined with flaA RFLP provided a good method for ascertaining the relatedness of strains isolated from carcasses and caecal contents. This study showed that carcass contamination is related to the within-flock prevalence of campylobacter colonisation, but that contamination from previously processed flocks was also significant, especially on carcasses from low prevalence flocks. Forced dry air cooling of carcasses reduced contamination levels.

RAPD PCR Type of Campylobacter Isolates from Cecum Contents of Broiles Chickens

In this investigation we typed Campylobacter isolates by RAPD PCR from the cecum contents of broiler chickens obtained at a slaughterhouse in Shizuoka prefecture. Campylobacter was isolated from 91 (56.9%) out of 160 birds from five out of eight flocks . Of the five Campylobacter positive flocks, three flocks possessed multiple RAPD types of Campylobacter, and two flocks possessed a single type. In the flocks where multiple types of Campylobacter were obtained, all the birds were positive and multiple RAPD types of Campylobacter were isolated from individual broiler samples. In these flocks, multiple episodes of Campylobacter contamination may have occurred . On the other hand, the three Campylobacter negative flocks were considered to have had no Campylobacter contaminations . The differences in Campylobacter colonization of the flocks in this study may, at least in part, reflect the differences in the hygiene measures of the broiler houses in which these birds are raised.

Distribution of Campylobacter spp. in Selected U.S. Poultry Production and Processing Operations

A study was conducted of 32 broiler flocks on eight different farms, belonging to four major U.S. producers. The farms were studied over 1 complete calendar year. Overall, 28 (87.5%) of the flocks became Campylobacter positive, and only four (12.5%) remained negative throughout the 6- to 8-week rearing period. In the majority of flocks, sampled every 2 weeks throughout production, Campylobacter-positive fecal and cecal samples were not detected until 4 to 8 weeks of age. In only six of the flocks were environmental samples found to be positive before shedding of Campylobacter was detected in the birds. Even in some of the Campylobacter-negative flocks, contamination of the rearing environment was positive for Campylobacter but did not result in the birds subsequently excreting the organism. These findings are discussed in relation to U.S. husbandry practices and present uncertainty about sources of Campylobacter infection for poultry flocks. Birds were often transported to the processing plant in coops that were already contaminated with Campylobacter, and the organisms were sometimes found in samples of scald water and chill water. After chilling, the proportions of Campylobacter-positive carcasses from different producers ranged from 21.0 to 40.9%, which is lower than in other studies, and possible reasons are considered.

Poultry as a source of Campylobacter and related organisms

Approximately 80% of raw chickens sold in the UK are contaminated with thermophilic campylobacters and they can be found on carcasses at levels as high as several thousand per cm 2 of skin. Thus, although they do not multiply on meat, they have a low infectious dose, so that human infection from undercooked meat, or as a result of handling raw poultry, is common. The precise contribution of poultry to human infection is not clear, but comparisons of types infecting humans and animals indicate that there are other important sources of human infection, particularly cattle. Rates of contamination of raw poultry meat with Arcobacter species and Helicobacter pullorum are also very high. Thermophilic campylobacters and H. pullorum colonize the chicken gut and rarely cause disease in poultry. Arcobacter species often appear to be environmental contaminants rather than part of the natural gut flora of poultry, although A. butzleri, in particular, can cause intestinal infections and abortion in humans. Thermophilic campylobacters arc not generally thought to be transmitted vertically via eggs, nor via feed or litter, provided rearing houses are cleaned and disinfected between flocks, and litter renewed. Flocks usually become infected at about 3 weeks of age. Every bird is usually rapidly colonized, with high levels (10 6 -10 7 cfu/g) in the caccal contents. The source of infection can be via unchlorinated water, but in situations where the water supply is not to blame, the precise source of infection is seldom identified. Infection could be via wild birds, rodents, or from farm operatives via boots or clothing. Infection has sometimes been associated with 'thinning' of flocks about a week prior to slaughter. Avoidance of infection during rearing therefore relies mostly on careful attention to hygiene, exclusion of vermin and a clean water supply. During transport, slaughter and dressing, Campylobacter-negative flocks can readily be contaminated from positive flocks. Contamination can be reduced by improved disinfection of transport crates, slaughter of uninfected flocks prior to infected flocks, and by careful attention to major points of cross-contamination on the line. A more effective measure would be to use a terminal decontamination step, such as trisodium phosphate, lactic acid, reduced pressure steam or gamma irradiation.

Changes in the carriage of Campylobacter strains by poultry carcasses during processing in abattoirs.

The recent development of simple, rapid genotyping techniques for Campylobacter species has enabled investigation of the determinative epidemiology of these organisms in a variety of situations. In this study we have used the technique of fla typing (PCR-restriction fragment length polymorphism analysis of the flaA and flaB genes) to identify the sources of strains contaminating the carcasses of five campylobacter-positive and two campylobacter-negative broiler flocks during abattoir processing. The results confirmed that, in the United Kingdom, individual broiler flocks are colonized by a limited number of subtypes of Campylobacter jejuni or C. coli. In some but not all cases, the same subtypes, isolated from the ceca, contaminated the end product as observed in carcass washes. However, the culture methodology, i.e, use of direct plating or enrichment, affected this subtype distribution. Moreover, the number of isolates analyzed per sample was limited. fla typing also indicated that some campylobacter subtypes survive poultry processing better than others. The extent of resistance to the environmental stresses during processing varied between strains. The more robust subtypes appeared to contaminate the abattoir environment, surviving through carcass chilling, and even carrying over onto subsequent flocks. From these studies it is confirmed that some campylobacter-negative flocks reach the abattoir but the carcasses from such flocks are rapidly contaminated by various campylobacter subtypes during processing. However, only some of these contaminating subtypes appeared to survive processing. The sources of this contamination are not clear, but in both negative flocks, campylobacters of the same subtypes as those recovered from the carcasses were isolated from the crates used to transport the birds. In one case, this crate contamination was shown to be present before the birds were loaded.