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Whole-Genome Sequencing has the potential to be an effective method for surveillance of foodborne diseases. This study aims to determine the genetic relatedness and prevalence of virulence-associated genes and antimicrobial resistance determinants in 135 Campylobacter jejuni , seven Campylobacter coli and three Campylobacter lari isolates from the poultry supply chain and a hospital in Spain. The isolates showed a wide genetic diversity between and within species with Clonal Complex 21 the most frequent lineage found. Among species, C. jejuni showed the highest prevalence of virulence genes (287/333) in which a high occurring variability was observed in the capsule biosynthesis and transport, O-linked flagellar glycosylation and lipooligosaccharide biosynthesis loci, with a great impact of phase-variation that led to 72 different virulence gene patterns among all isolates. High prevalence (> 90 %) of bla OXA -type β-lactamase genes and mutations in DNA gyrase gene associated with fluoroquinolones resistance were observed, and at a frequency similar to the tet(O) gene in C. jejuni (93 %) and C. coli (86 %), both of which also harboured resistance determinants to aminoglycosides with a higher occurrence rate in C. coli (43 %), that was the only species in which mutations in the 23S ribosomal subunit conferring resistance to erythromycin were identified (43 %). The present study constitutes the largest genomic survey of Campylobacter isolates in Spain providing insight into the prevalence and diversity of the pathogen along the poultry supply chain in the country. • Large genetic diversity exists among Campylobacter isolates from Spain. • Campylobacter spp. show a complex combination of virulence factors. • Phase variation contributes to the diversity of virulence profiles in Campylobacter. • High rates of antibiotic resistant genes exist in C. jejuni and C. coli. [ABSTRACT FROM AUTHOR]

The Hazard Analysis and Critical Control Point (HACCP) system plays a crucial role in ensuring food safety within food service establishments, effectively reducing the risk of foodborne diseases. This study focused on assessing the risk of microbe contamination in poultry-based cook-served food during meal preparation in four restaurants and five selected HACCP-certified hotels in eastern China. We examined samples collected from 26 poultry-based cooked dishes, 248 food contact surfaces, 252 non-food contact surfaces, and 121 hand swabs. Our findings indicated a favorable trend of compliance with Chinese national standards, as Escherichia coli and Campylobacter were not detected in any cooked food samples. However, the microbiological assessments revealed non-compliance with total plate count standards in 7 % of the cooked samples from restaurants. In contrast, both dine-in hotels and restaurants exhibited significant non-compliance with guidance concerning food and non-food contact surfaces. Furthermore, our study found that chefs' hand hygiene did not meet microbiological reference standards, even after washing. Notably, Campylobacter persisted at 27 % and 30 % on chefs' hands, posing a significant risk of cross-contamination and foodborne diseases. These findings emphasize the urgent necessity for enhanced supervision of hygiene procedures and process monitoring in the HACCP-certified establishments engaged in the preparation and serving of food. Targeted interventions and food safety education for different chef subgroups can enhance food handling practices and reduce the risk of foodborne diseases in independent food establishments. • This study aimed to assess microbe contamination risk in food establishments in China. • Escherichia coli and Campylobacter were not detected in any of the cooked food samples. • Food establishments exhibited critical non-compliance with guidelines for kitchen surfaces. • Chefs' hand hygiene might possess a huge risk of cross-contamination and foodborne diseases. • Targeted interventions and education are needed to improve the awareness of safe handle practices. [ABSTRACT FROM AUTHOR]

Campylobacter was considered asaccharolytic, but is now known to carry saccharide metabolization pathways for L-fucose and d -glucose. We hypothesized that these clusters are beneficial for Campylobacter niche adaptation and may help establish human infection. We investigated the distribution of d -glucose and L-fucose clusters among ∼9600 C. jejuni and C. coli genomes of different isolation sources in the Netherlands, the United Kingdom, the United States of America and Finland. The L-fucose utilization cluster was integrated at the same location in all C. jejuni and C. coli genomes, and was flanked by the genes rpoB , rpoC , rspL , repsG and fusA, which are associated with functions in transcription as well as translation and in acquired drug resistance. In contrast, the flanking regions of the d -glucose utilization cluster were variable among the isolates, and integration sites were located within one of the three different 16S 23S ribosomal RNA areas of the C. jejuni and C. coli genomes. In addition, we investigated whether acquisition of the L-fucose utilization cluster could be due to horizontal gene transfer between the two species and found three isolates for which this was the case: one C. jejuni isolate carrying a C. coli L-fucose cluster, and two C. coli isolates which carried a C. jejuni L-fucose cluster. Furthermore, L-fucose utilization cluster alignments revealed multiple frameshift mutations, most of which were commonly found in the non-essential genes for L-fucose metabolism, namely, Cj0484 and Cj0489. These findings support our hypothesis that the L-fucose cluster was integrated multiple times across the C. coli / C. jejuni phylogeny. Notably, association analysis using the C. jejuni isolates from the Netherlands showed a significant correlation between human C. jejuni isolates and C. jejuni isolates carrying the L-fucose utilization cluster. This correlation was even stronger when the Dutch isolates were combined with the isolates from the UK, the USA and Finland. No such correlations were observed for C. coli or for the d -glucose cluster for both species. This research provides insight into the spread and host associations of the L-fucose and d -glucose utilization clusters in C. jejuni and C. coli , and the potential benefits in human infection and/or proliferation in humans, conceivably after transmission from any reservoir. • L-fucose cluster integration events across C. coli and C. jejuni phylogeny are shown. • L-fucose cluster integration is conserved, but that of the d -glucose utilization cluster is not. • Human C. jejuni isolates and isolates with the L-fucose utilization cluster correlate. • L-Fucose metabolism capacity provides selective advantage in C. jejuni transmission. [ABSTRACT FROM AUTHOR]

Filth flies, especially house flies, can harbor and disseminate human pathogens to food and food contact surfaces. To determine the potential of flying insects to carry Salmonella and Campylobacter from poultry grow out houses, a total of 2,164 flying insects were caught and segregated based on flying insect family type and farm location in the Upstate, Middle, and Coastal parts of South Carolina in the United States for two 14-d sampling periods at 3 separate farms. Captured flying insects included house flies in the family Muscidae inside the poultry house (N = 289), house flies just outside the poultry house (N = 1023), and house flies 100 meters from the poultry houses (N = 547). Other flying insects included wasps in the family Vespidae species (spp.) captured just outside the poultry house (N = 71), Vespidae 100 meters from the poultry house (N = 126), flesh flies in the family Sarcophagidae just outside the poultry house (N = 13), and flesh flies 100 meters from the poultry house (N = 9), blow flies in the family Calliphoridae 100 meters from the poultry house, darkling beetles in the family Tenebrionidae just outside the poultry house (N = 30), and darkling beetles 100 meters from the poultry house (N = 56). Populations of Campylobacter spp., Salmonella spp., and total aerobic organisms (APC) were recovered from flying insects as well as the number of Salmonella spp. and Campylobacter spp. positive flying insects at a 100 m distance from the broiler farms. Along with insect groups, chicken feces in the grow-out houses from 3 farms, cow manure around farm 1 and farm 2, and dog feces around farm 1 were also sampled. While no Campylobacter jejuni was recovered from any of the samples, including flying insect groups, chicken feces, cow manures, and dog feces, Campylobacter coli positive samples were detected in the cow manure samples in both collection periods, 100m-Calliphoridae, out-house flies and 100m-darkling beetles in 1 out of 2 collection periods on farm 2. Moreover, positive Serogroup B Salmonella spp. were detected in the groups in-chicken feces, in- house flies, and out- house flies on farm 2 and positive Serogroup C Salmonella spp. were detected in the groups of in- chicken feces, out- house flies, and 100m- house flies on farm 3. These data suggest that house flies may be a vector in the transmission of Salmonella spp. to and from broiler farms. [ABSTRACT FROM AUTHOR]

• Infectious diarrhea was assessed in 140 allogeneic hematopoietic cell recipients. • The cumulative incidence of at least one episode after 1 year was 32%. • The incidence rate was the highest in the pre-engraftment phase. • Most frequently observed were Clostridioides difficile, adenovirus, Enteropathogenic Escherichia coli and Campylobacter jejuni. To determine the incidence of infectious diarrhea after allogeneic hematopoietic cell transplantation (HCT) using a multiplex polymerase chain reaction assay and assess risk factors for developing infectious diarrhea. This was a single-center retrospective study of 140 consecutive allogeneic HCT recipients. Infectious diarrhea was assessed using a laboratory-developed multiplex polymerase chain reaction the first year after transplantation. The incidence rate of infectious diarrhea episodes was 47 per 100 person-years, with the highest rate observed in the pre-engraftment phase. Most episodes were seen as nosocomial infections (38%) and most affected patients (82%) had only one episode of infectious diarrhea. The cumulative incidence of at least one episode of infectious diarrhea was 32% after 1 year. Nonrelapse mortality was higher in transplant recipients with at least one episode of infectious diarrhea (hazard ratio (HR) 2.02, 95% CI = 1.07-3.80). The most frequently observed pathogens were Clostridium difficile , adenovirus, Enteropathogenic Escherichia coli, and Campylobacter jejuni. Patients with acute lower gastrointestinal graft-vs-host disease stage 3 or 4 (HR 3.68, 95% CI = 1.57-8.63) conferred a higher risk for a first infectious diarrhea episode. Infectious diarrhea after allogeneic HCT was seen in about one-third of the patients, mostly as nosocomial infection in the pre-engraftment phase. [ABSTRACT FROM AUTHOR]

• Animals had the highest pooled prevalence estimate (PPE) of Campylobacter infection. • The majority of Campylobacter prevalence studies were conducted in animals. • The highest antibiotic resistance PPE by Campylobacter isolates is against clindamycin. • The combined multidrug resistance PPE of Campylobacter isolates is 35.3%. This study investigated the prevalence and antibiotic resistance (AR) profiles of Campylobacter spp. isolated from animals, humans, and the environment in South Africa based on available published data. Original articles published from January 1, 1990 to January 1, 2021 were searched from PubMed, ScienceDirect, Google Scholar, Africa Index Medicus, Scopus, and African Journal Online databases. Data were analyzed with Comprehensive Meta-Analysis (version 3.0). After screening, articles on animals (n = 25), humans (n = 7), environment (n = 3), animals/environment (n = 2), and a (n = 1) study on animals, humans, and the environment were included in this review. The pooled prevalence estimates (PPEs) were 28.8%, 16.4%, and 28.4% in animals, humans, and the environment, respectively. The Campylobacter jejuni and Campylobacter coli species were commonly isolated from humans, animals, and the environment in South Africa. The AR profiles were screened from 2032 Campylobacter spp., with the highest PPE of AR observed against clindamycin (76.9%) and clarithromycin (76.5%). Campylobacter isolates tested with the disk diffusion assay and minimum inhibitory concentration methods recorded an overall AR prevalence of 35.3% and 37.1%, respectively, whereas multidrug resistance PPE was 35.3%. Regular surveillance of Campylobacter spp. prevalence and its antimicrobial resistance strains is recommended, as well as the formulation of a "One Health" approach for better management and control of Campylobacter spp. infection in South Africa. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine prevalence and perform genomic analysis of Salmonella spp. and Campylobacter spp. isolated from different stages of an integrated NAE broiler complex. Environmental samples were screened with 3M-Molecular Detection System (MDS) and MDS positive samples were further processed for confirmation of results and identification. Core genome-based phylogenies were built for both bacteria isolated from this study along with selected NCBI genomes. The odds ratios and 95% confidence limits were compared among stages and sample types (α < 0.05) using multivariable model. Based on MDS results, 4% and 18% of total samples were positive for Salmonella spp. and Campylobacter spp. respectively. The odds of Salmonella detection in hatchery samples were 2.58 times as likely as compared to its detection in production farms' samples (P = 0.151) while the odds of Campylobacter detection in production farms' samples were 32.19 times as likely as its detection in hatchery (P = 0.0015). Similarly, the odds of Campylobacter detection in boot swabs, soil, water, and miscellaneous samples were statistically significant (P < 0.05) as compared with fly paper as reference group. The serovars identified for Salmonella were Typhimurium, Barranquilla, Liverpool, Kentucky, Enteritidis, Luciana, and Rough_O:r:1,5. For Campylobacter, the species identified were Campylobacter jejuni and Campylobacter coli. Phylogeny results show close genetic relatedness among bacterial strains isolated from different locations within the same stage and between different stages. The results show possibility of multiple entry points of such bacteria entering broiler complex and can potentially contaminate the final raw product in the processing plant. It suggests the need for a comprehensive control strategy with strict biosecurity measures and best management practices to minimize or eliminate such pathogens from the poultry food chain. [ABSTRACT FROM AUTHOR]

[Display omitted] • Multiple pathogens and antimicrobial resistance genes in wastewater were quantified. • Freezing of wastewater and the biomass concentration method affected virus quantity. • Patterns in bacterial pathogen quantities followed seasonal disease trends. • Clinically relevant beta-lactamase genes were ubiquitous in Finnish wastewaters. • Resistome abundances revealed geographical but not seasonal differences. Wastewater-based surveillance (WBS) of infectious disease agents is increasingly seen as a reliable source of population health data. To date, wastewater-based surveillance efforts have largely focused on individual pathogens. However, given that wastewater contains a broad range of pathogens circulating in the population, a more comprehensive approach could enhance its usability. We focused on the simultaneous detection of SARS-CoV-2, sapovirus, Campylobacter jejuni , Campylobacter coli , Salmonella spp., pathogenic Escherichia coli , C ryptosporidium spp., Giardia spp. and antimicrobial resistance genes (ARGs) of clinical relevance. To achieve this goal, biomass concentration and nucleic acid extraction methods were optimized, and samples were analyzed by using a set of (RT)-qPCR and (HT)-qPCR methods. We determined the prevalence and the spatial and temporal trends of the targeted pathogens and collected novel information on ARGs in Finnish wastewater. In addition, the use of different wastewater concentrates, namely the ultrafiltered concentrate of the supernatant and the centrifuged pellet, and the effect of freezing and thawing wastewater prior to sample processing were investigated with the indicator microbe crAssphage. Freeze-thawing of wastewater decreased the gene copy count of crAssphage in comparison to analyzing fresh samples (p < 0.001). Campylobacters were most abundant in two of the four studied summer months (30 % detection rate) and in wastewaters from regions with intensive animal farming. Salmonella , however, was detected in 40 % of the samples without any clear seasonal trends, and the highest gene copy numbers were recorded from the largest wastewater treatment plants. Beta-lactamase resistance genes that have commonly been detected in bacteria isolated from humans in Finland, namely bla CTX-M , bla OXA48 , bla NDM , and bla KPC , were also frequently detected in wastewaters (100, 98, 98, and 70 % detection rates, respectively). These results confirm the reliability of using wastewater in public health surveillance and demonstrate the possibility to simultaneously perform WBS of multiple pathogens. [ABSTRACT FROM AUTHOR]

Campylobacter is a bacterial pathogen that causes human foodborne illnesses worldwide, and outbreaks have been associated with consumption of undercooked chicken livers. The objectives of this study were to compare two PCR assays of 250 Campylobacter isolates for identification to species, to assess antibiotic resistance of the isolates, and to analyze genetic diversity of the quinolone resistance determining regions (QRDRs) of the isolates. A double-blind design was used to identify the species of Campylobacter; 181 (72%) of the isolates were identified as Campylobacter jejuni, and 69 (28%) isolates were identified as Campylobacter coli by both PCR assays. A total of 93 (37.2%) isolates were resistant to at least one antibiotic. Among 88 C. jejuni isolates, 33 (18%) were resistant to nalidixic acid (NAL) and ciprofloxacin (CIP), 25 (14%) were resistant to tetracycline (TET), and 18 (10%) were resistant to NAL and TET. Two C. jejuni isolates were resistant to four of the tested antibiotics, and one isolate was resistant to five antibiotics. Two C. coli isolates were resistant to TET, and two were resistant to NAL, CIP, and TET. The amino acid sequences of the QRDRs for the isolates had eight point mutations and could be classified into 12 groups. Thirty-eight C. jejuni isolates resistant to NAL and CIP had a point mutation at residue 86 (substitution from threonine to isoleucine). However, six isolates without this substitution were resistant to NAL and/or CIP. Ten isolates with a point mutation at residue 86 were susceptible to NAL and CIP. This observation suggests that in addition to the substitution at residue 86 other mechanisms may confer resistance to quinolones. Further studies are needed to elucidate mechanisms for quinolone resistance in Campylobacter. The Campylobacter spp. isolated from chicken livers in this study were resistant to quinolones and other classes of antibiotics. [ABSTRACT FROM AUTHOR]

Campylobacter spp., such as Campylobacter jejuni and Campylobacter coli , are important zoonotic Gram-negative pathogens that cause acute intestinal diseases in humans. In this study, a retrospective analysis was conducted on previously collected Campylobacter isolates from antimicrobial resistance surveillance. A total of 29 optrA -positive C. coli strains were identified and subjected to second-generation sequencing. Multilocus sequence typing and single nucleotide polymorphism analyses demonstrated that the 29 optrA -positive isolates were genetically homogeneous. Notably, among the 29 isolated strains, the Δ optrA variants exhibit a nonsense mutation at position 979 where the base C is substituted by T, leading to the formation of a premature termination codon. The alignment of sequences and genetic environmental characteristics suggested that Δ optrA located on a chromosomally carried multidrug-resistant genomic island. There are other resistant genes on the multidrug resistance genomic island, such as aph(2'')-If , aph(3′)-III , aadE , tet (O), tet (L), cat , erm (A), optrA and bla OXA-61. As a result, the 29 Δ optrA -positive strains displayed susceptibility to both florfenicol and linezolid. The Δ optrA gene is linked to the erm (A) gene, resulting in the formation of translocatable unit (TU) that are encompassed by two copies of IS 1216 mobile elements. Multiple occurrences of similar TUs have been documented in numerous C. coli and provided evidence for the significance of TUs in facilitating the transfer of drug resistance genes in C. coli. • The Δ optrA gene is located on the MDRGI of C. coli. • All the Δ optrA -positive C. coli strains belonged to ST872. • One transposition unit was formed through the involvement of the IS 1216 element. • C. coli integrates various resistance genes, leading to the formation of MDRGI. [ABSTRACT FROM AUTHOR]

Campylobacter jejuni and Campylobacter coli are the leading causes of bacterial intestinal infections worldwide, while Campylobacter fetus subsp. fetus (C. fetus) has been reported to cause extraintestinal infections, including medical device implant infections. However, breast implant infections have rarely been reported. We describe the case of a 64-year-old woman with breast implant infection and vertebral osteomyelitis due to C. fetus. The patient recovered by surgical removal of the infected left implant and was treated with antibiotics for 6 weeks. However, two weeks after the completion of antibiotics, she experienced an infection in the right implant due to C. fetus , which had developed quinolone resistance with a G91T mutation during the treatment course. This case showed that C. fetus can cause breast implant infections, and although the infection may appear to be unilateral initially, the possibility of sequential contralateral infection should be considered. [ABSTRACT FROM AUTHOR]

A pilot survey was performed to determine the prevalence of Campylobacter jejuni and Campylobacter coli on three age classes (lamb, hogget, and mutton) of ovine carcass trim postdressing and prechill. Sampling of hogget carcasses was undertaken 6 months before sampling of lamb and mutton carcasses. A total of 120 trim samples were collected from 11 processing plants across New Zealand. All samples were enriched and screened using PCR for the presence of C. jejuni and C. coli, and isolation was attempted for all screen-positive samples. Enumeration of Campylobacter from lamb trim samples showed that Campylobacter bacteria were present in very low numbers (< 10 CFU/g). The overall prevalence of Campylobacter for ovine trim based on PCR detection was 33% (39 of 120 samples), with prevalences for hogget, lamb, and mutton carcass trim of 56% (28 of 50), 11% (4 of 35), and 20% (7 of 35), respectively. Whole genome sequencing was performed on a selection of C. jejuni and C. coli isolates, and the data were used to subtype using multilocus sequence typing (MLST) and whole genome MLST. Twenty-five MLST sequence types (STs) were identified among 44 isolates, including ST42, ST50, ST3222, and ST3072, which have been previously reported to be associated with ruminant sources. Four novel STs were also identified. Whole genome MLST analysis further discriminated isolates within a single ST type and demonstrated a genetic diversity among the ovine isolates collected. Genes associated with the oxacillinase class of β-lactamase enzymes were identified in 41 of 44 Campylobacter isolates. This study provides preliminary data that can be incorporated into existing source attribution models to assist in determining the potential contribution of ovine sources to the burden of campylobacteriosis in New Zealand. Campylobacter prevalence for ovine carcass trim based on PCR detection was 33%. Enumeration of Campylobacter bacteria was < 10 CFU/g on lamb trim. Twenty-five multilocus sequence types (STs) were identified among 44 isolates. Whole genome sequence analysis discriminated isolates within a single ST. [ABSTRACT FROM AUTHOR]

In vivo and in vitro assessment of commercial probiotic and organic acid feed additives in broilers challenged with Campylobacter coli. [Extracted from the article]

Chicken is a leading source of thermotolerant Campylobacter, which triggers human foodborne enteritis. This study evaluated thermotolerant Campylobacter contamination of retail chicken in southern Brazil, using qualitative and quantitative analyses. Selective enrichment in Bolton broth for 24 and 48 h after plating onto modified charcoal–cefoperazone– deoxycholate (mCCD) agar and Preston agar was assessed. The combined results of the detection and enumeration methods revealed a frequency of 70% occurrence of thermotolerant Campylobacter in chicken samples. Campylobacter was enumerated in 60% of the samples, whereas 46% of the samples were positive in the qualitative analysis. Quantitative analysis showed average counts of 3.10 6 0.15 log10 CFU/sample. Higher numbers of Campylobacter-positive samples were found using 24-h enrichment before plating onto Preston agar (46%) than onto mCCD agar (2%). The majority of isolated strains were identified as Campylobacter jejuni, and Campylobacter coli was also found but to a lesser extent. Subtyping revealed a clear distinction between strains isolated from different chicken sources. The enriched samples plated onto mCCD agar showed extensive spreading of nonproducing extended-spectrum b-lactamases Proteus mirabilis that hampered the identification of Campylobacter colonies. P. mirabilis strains showed resistance to cefoperazone, trimethoprim, and polymyxin B present in broth and plate media used and were inhibited by rifampicin present in Preston agar. The results underline the effect of the spread of contaminant strains on Campylobacter cultures, which might be prevented using a recently revised International Organization for Standardization method for qualitative analysis of chicken. [ABSTRACT FROM AUTHOR]

Campylobacter is among the most frequent agents of bacterial gastroenteritis in Europe and is primarily linked to the consumption of contaminated food. The aim of this study was to assess genomic diversity and to identify antimicrobial resistance and virulence genes of 155 Campylobacter isolated from broiler carcasses (neck skin samples) in a large-scale Swiss poultry abattoir over a three-year period. Samples originated from broilers from three different types of farming systems (particularly animal-friendly stabling (PAFS), free-range farms, and organic farms). Campylobacter jejuni (n = 127) and Campylobacter coli (n = 28) were analysed using a whole genome sequencing (WGS) approach (MiniSeq; Illumina). Sequence types (STs) were determined in silico from the WGS data and isolates were assigned into complex types (CTs) using the cgMLST SeqSphere+ scheme. Antimicrobial resistance genes were identified using the Resistance Gene Identifier (RGI), and virulence genes were identified using the virulence factor database (VFDB). A high degree of genetic diversity was observed. Many sequence types (C. jejuni ST19, ST21, ST48, ST50, ST122, ST262 and C. coli ST827) occurred more than once and were distributed throughout the study period, irrespective of the year of isolation and of the broiler farming type. Antimicrobial resistance determinants included bla OXA and tet (O) genes, as well as the T86I substitution within GyrA. Virulence genes known to play a role in human Campylobacter infection were identified such as the wlaN, cstIII , neuA1, neuB1, and neuC1. Subtyping of the Campylobacter isolates identified the occurrence of a highly clonal population of C. jejuni ST21 that was isolated throughout the three-year study period from carcasses from farms with geographically different locations and different farming systems. The high rate of genetic diversity observed among broiler carcass isolates is consistent with previous studies. The identification of a persisting highly clonal C. jejuni ST21 subtype suggests that the slaughterhouse may represent an environment in which C. jejuni ST21 may survive, however, the ecological reservoir potentially maintaining this clone remains unknown. • C. jejuni and C. coli isolated from broiler carcasses are genetically diverse by cgMLST. • Antimicrobial resistance genes comprise bla OXA , tet (O) and GyrA T86I. • A highly clonal population of C. jejuni ST21 CT2570 persisted throughout the 3-year study period. • The majority of C. jejuni co-harbouring the virulence factors cstIII neuA1, neuB1, and neuC1 belonged to ST21. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the prevalence and distribution of Campylobacter species in a variety of fresh and frozen meat and offal products collected from retail outlets in New South Wales (NSW), Queensland (Qld), and Victoria (Vic). A total of 1,490 chicken, beef, lamb, and pork samples were collected from Australian supermarkets and butcher shops over a 2-year sampling period (October 2016 to October 2018). Campylobacter spp. were detected in 90% of chicken meat and 73% of chicken offal products (giblet and liver), with significantly lower prevalence in lamb (38%), pork (31%), and beef (14%) offal (kidney and liver). Although retail chicken meat was frequently contaminated with Campylobacter, the level of contamination was generally low. Where quantitative analysis was conducted, 98% of chicken meat samples, on average, had <10,000 CFU Campylobacter per carcass, with 10% <21 CFU per carcass. Campylobacter coli was the most frequently recovered species in chicken meat collected in NSW (53%) and Vic (56%) and in chicken offal collected in NSW (77%), Qld (59%), and Vic (58%). In beef, lamb, and pork offal, C. jejuni was generally the most common species (50 to 86%), with the exception of pork offal collected in NSW, where C. coli was more prevalent (69%). Campylobacter prevalence was significantly higher in fresh lamb (46%) and pork (31%) offal than in frozen offal (17 and 11%, respectively). For chicken, beef, and pork offal, the prevalence of Campylobacter spp. was significantly higher on delicatessen products compared with prepackaged products. This study demonstrated that meat and offal products are frequently contaminated with Campylobacter. However, the prevalence is markedly different in different meats, and the level of chicken meat portion contamination is generally low. By identifying the types of meat and offal products types that pose the greatest risk of Campylobacter infection to consumers, targeted control strategies can be developed. [ABSTRACT FROM AUTHOR]

Campylobacter jejuni and Campylobacter coli originating from poultry meat have been the most important causes of foodborne bacterial gastroenteritis in the European Union since 2005. In-feed application of maternal antibodies from vaccinated hens was shown to confer protection of broilers against Campylobacter infection. Here, it was investigated if these vaccines can be used to protect broilers against Campylobacter infection after in ovo vaccination. Embryos were immunized in ovo at day 18 with a bacterin or a subunit vaccine and at 19 D post hatch, these birds were inoculated with C. jejuni according to a seeder model. Quantification of C. jejuni in the broilers cecal content showed that the in ovo vaccinated birds were not protected against C. jejuni infection. Quantification of blood anti- Campylobacter antibody titers did not show any induction of Campylobacter -specific serological response in the vaccinated birds, which may explain the lack of protection in the vaccinated chicks. [ABSTRACT FROM AUTHOR]

• Development of a single step ELIME assay for the detection of Campylobacter jejuni and coli. • Exploitation of different antibody combinations. • Evaluation of the antibody reactivity towards Campylobacter, as a function of the culture medium and cell treatment. • Selectivity study versus different C. jejuni strains and other non-target microorganisms. Campylobacter is an important cause of acute bacterial disease in human worldwide. Within the genus Campylobacte r, C. jejuni and C. coli are the two species responsible for over 95% of human infections. The purpose of this work is the development of an Enzyme-Linked-Immuno-Magnetic-Electrochemical (ELIME) assay for a simple and rapid detection of both Campylobacte r species. Tosyl-activated magnetic beads (MBs), supporting a sandwich immunological complex, were coupled to a strip of 8-magnetized screen-printed electrodes, as electrochemical transducing element. With the aim to simultaneously detect C. jejuni and C. coli, four anti-campylobacter antibodies were screened in ELISA paying the attention to the selection of both culture medium and cell treatment (whole campylobacter cells inactivated with NaN 3 and heat killed cells) for a suitable antigen-antibody interaction. Six combinations of the selected anti-campylobacter antibodies, unconjugated and conjugated with Horseradish Peroxidase (HRP) or Alkaline Phosphatase (AP), were exploited as sensing strategy to set-up the ELIME assay. The antibody pair, consisting of a goat polyclonal antibody anti-campylobacter (coated on the surface of MBs) and the same antibody conjugated with AP (to label the captured cells), was selected given the best response for both Campylobacter species and the lowest detection limit (9 × 103 CFU/mL for C. jejuni and 2 × 104 CFU/mL for C. coli). The selectivity of the ELIME assay was proved by analyzing other C. jejuni strains and non target microorganisms attaining promising results. [ABSTRACT FROM AUTHOR]

Seasonal variation in the prevalence and antimicrobial resistance of Campylobacter species in the Ethiopian dairy value chain was investigated. Dairy food samples (456) were collected in the dry and wet seasons in three regions of Ethiopia. Campylobacter species were detected in 20 % of samples collected in the wet season. The overall prevalence did not differ significantly between the wet and dry seasons. However, in the Oromia region, there was a 5 times greater chance of finding Campylobacter species in milk and milk products during the wet season than in the dry season. Among Campylobacter -positive samples collected countrywide, 89 % were contaminated with Campylobacter jejuni , and 11 % with Campylobacter coli. In the dry season, all Campylobacter -positive samples were contaminated with C. jejuni. Most Campylobacter species were resistant to tetracycline (89 %), followed by erythromycin (74 %), and ciprofloxacin (57 %); 43 % of the isolates were resistant to more than two drugs from two different classes. [ABSTRACT FROM AUTHOR]

In 2012, the European Food Safety Authority (EFSA) proposed harmonised epidemiological indicators (HEIs) for poultry at different production stages: Salmonella , Campylobacter , extended-spectrum β-lactamase/AmpC β-lactamase (ESBL/AmpC) producing Enterobacteriaceae and generic Escherichia (E.) coli. The HEIs are based on existing monitoring systems or the sampling strategies provided by EFSA. To realise the full benefit of HEIs, risk managers should use them for farm and abattoir categorisation and for adapting the existing meat inspection systems. As HEIs are not legal requirements within the European Union (EU), it is unclear which HEIs are used in which country, to date. Therefore, an online survey was conducted in Europe to gather knowledge about the implementation, application and consequences following on from the HEIs in existing official and private monitoring and surveillance systems (MOSS). A total of 34 answer sets from participants working in the framework of official surveillance or as food business operators in broiler abattoirs were collected from eleven EU member states (EU-MS) and four non-EU countries. While all participants stated that testing for Salmonella is performed, HEI 4- Salmonella , which corresponds to the process hygiene criterion (PHC) for Salmonella was applied by 62% of the participants. In total, 94% of the participants reported that they test for Campylobacter. Among them, 71% stated that testing is performed for HEI 5- Campylobacter , which corresponds to PHC for Campylobacter. Although testing neck skin samples for Salmonella and Campylobacter after chilling are official and mandatory MOSS in the EU, not all participants from EU-MS (Salmonella : 6/11 EU-MS; Campylobacter : 8/11 EU-MS) confirmed to comply with. Altogether, 56% of the participants (from 6 EU-MS and 2 non-EU countries) stated that they test for E. coli. Ten of them reported that the testing is performed at the abattoir after chilling according to the suggested HEI for generic E. coli as a hygiene indicator. Consequences that result from the existing MOSS for the three examined pathogens (Salmonella , Campylobacter and E. coli) were mainly rising awareness, farm risk categorisation and feedback to the farmer. According to the answers from the participants, the HEIs suggested by EFSA for broilers are currently implemented in most EU-MS. One reason could be that some of the according MOSS are required by EU law. As intended by EFSA, the participants stated that they use HEIs for farm risk categorisation as one of the three top consequences following from MOSS for the three mentioned pathogens. For improving the knowledge and application of HEIs in the context of risk-based meat safety assurance systems, specific training could be helpful. • Implementation of HEIs at different production stages varies between countries. • HEIs for broilers are implemented in most EU member states. • Main consequences are raising awareness, farm risk categorisation, feedback to farmers. • Currently implemented monitoring/surveillance systems for broilers are appropriate. • Training is needed in HEI applications for risk-based meat safety assurance systems. [ABSTRACT FROM AUTHOR]

Salmonella and Campylobacter spp. are important foodborne pathogens of concern. Both horizontal and vertical transmission of Salmonella into poultry flocks is well documented to occur, whereas only horizontal transmission of Campylobacter has been confirmed. The objective of this study was to evaluate the ability of Salmonella and Campylobacter to translocate from egg contents into the embryo during incubation. Three experiments were performed comparing Salmonella recovery from eggs that were cold-stored (48 h) and incubated up to D15 or D18 of incubation. Four experiments were performed comparing Campylobacter recovery from eggs that were cold-stored and incubated up to D15 or D18 of incubation. Eggs were injected with 102-3 CFU Salmonella Enteritidis or 103 CFU Campylobacter coli into the albumen or yolk, and sampling of the egg contents and viable embryos was performed periodically during cold-storage and incubation. In the Salmonella experiments, egg contents from yolk injected eggs sampled during incubation were 100% positive, and no viable embryos were present. In contrast, Salmonella albumen injected eggs sampled on D18 had 51% positive egg contents and 37% positive embryos. In the Campylobacter experiments, 15% of embryos of yolk injected eggs were positive at D15, but no egg contents or embryos were positive at D18 of incubation. Results indicate that the albumen is likely to be the site for contamination than the yolk for forming eggs resulting in Salmonella positive embryos. Additionally, Campylobacter vertical transmission appears to be unlikely, but if it does occur, the location of the contamination is more likely the yolk contents. [ABSTRACT FROM AUTHOR]

• Vascular inflammation may be linked to Campylobacter coli infection. • Post-infectious vasculitis may cause relapsing fever after an acute infection. • 18F-fluorodeoxyglucose positron emission tomography/computed tomography may be helpful for the diagnosis of post-infectious vasculitis. [ABSTRACT FROM AUTHOR]

• Patients with agammaglobulinemia are prone to Campylobacter species infections. • Antibiotic resistance among Campylobacter is an emerging problem worldwide. • In Southeast Asia fluoroquinolone-resistant Campylobacter is a severe problem. • For macrolide- and fluoroquinolone-resistant isolates, oral aminoglycoside might be the most appropriate choice. Campylobacter is the most common cause of infectious diarrhea in agammaglobulinemia patients. These infections can be severe, prolonged, and recurrent in such patients. We report a 29-year-old male patient with X-linked agammaglobulinemia with Campylobacter coli enterocolitis that persisted for nine months despite multiple 10- to 14-day courses of oral ciprofloxacin and azithromycin. The isolate was highly resistant to ciprofloxacin, erythromycin, tetracycline, and fosfomycin. The patient failed to respond to intravenous ertapenem, 1.0 g/day for two weeks, to which the pathogen was susceptible. He was finally cured with oral gentamicin, 80 mg four times daily, and stool cultures remained negative during the seven-month follow-up. Oral aminoglycoside might be the most appropriate choice for eradication of persistent Campylobacter in the intestinal tract for macrolide- and fluoroquinolone-resistant isolate in agammaglobulinemia patients with chronic diarrhea or relapsing systemic infections. [ABSTRACT FROM AUTHOR]

Rapid and accurate identification of Campylobacter-positive broiler flocks and carcasses expedites separation and control interventions before release into the food supply chain and directly facilitates a reduction in the prevalence of human campylobacteriosis. In this study, the diagnostic performance of fluorescent loop-mediated isothermal amplification (LAMP) assays for the direct detection of Campylobacter jejuni and Campylobacter coli in broiler cloacal and cecal samples were evaluated and compared with that of turbidimetric LAMP approaches investigated previously. The duplex fluorescent LAMP assay had significantly higher (P < 0.05) diagnostic sensitivity (93.1%, 54 of 58 samples) than did the turbidimetric LAMP assay (82.8%, 48 of 58 samples) for detecting C. jejuni and C. coli in broiler cloacal samples, whereas the singleplex fluorescent LAMP assay had equivalent diagnostic sensitivity. For cecal samples, the diagnostic sensitivity of the fluorescent LAMP assay (100%, 38 of 38 samples) was the same as that of the turbidimetric LAMP. Fluorescent LAMP significantly reduced (P < 0.05) the maximum detection time for Campylobacter-positive cloacal and cecal samples to 28 and 11 min, respectively, and reduced the influence of amplification inhibitors responsible for most false-negative results obtained for cloacal samples with the turbidimetric LAMP assay. The diagnostic accuracy of the fluorescent LAMP assay for the direct detection of C. jejuni and C. coli in cloacal and cecal samples was 97.7 and 100%, respectively. These findings indicate that fluorescent LAMP assays are robust, highly accurate, and field-applicable methods for the identification of C. jejuni and C. coli, which will allow more accurate monitoring of food safety at various stages of the food supply chain at farms and slaughter facilities. [ABSTRACT FROM AUTHOR]

Campylobacter is the most common cause of infectious intestinal disease, with nearly all cases caused by two species: C. jejuni and C. coli . We recently reported a gold nanoparticle-based two-stage aptasensing platform, which was improved in the present study for the rapid and on-site detection of both C. jejuni and C. coli in food samples. Compared to the previous platform, the improved platform yielded a more obvious colour change from red to purple due to the aggregation of gold nanoparticles, and does not require additional time or a pH optimization step for the aptamers to be adsorbed onto the gold nanoparticles. Using a highly specific aptamer that binds to live C. jejuni and C. coli , the improved aptasensor was highly effective for testing pure culture samples. The accuracy of the newly developed platform was comparable ( p  = 0.688) to that of the gold-standard detection method of tazobactam-supplemented culture, whereas it was superior to the official agar-based detection method ( p  = 0.016) in a validation study with 50 naturally contaminated chicken carcass samples. This is the first study on a colorimetric sensor that targets both live C. coli and C. jejuni in naturally contaminated samples. In addition, we provide the first evidence that both morphological status and the amount of Campylobacter present play key roles in the effectiveness of colorimetric detection. Thus, suitable selection of an antibody or aptamer with consideration of the morphological status of pathogens in samples is essential for direct detection without enrichment. Our data suggest that the sensor developed in this study can provide an excellent screening method, with a reduction in the detection time from 48 h to 30 min after enrichment, thus saving time, labour, and cost. [ABSTRACT FROM AUTHOR]

It is important to determine whether valuable broiler-breeder chicks are contaminated with Campylobacter. It is important to have a non-destructive method to determine whether microorganisms such as Campylobacter are present without sacrificing the animal. The objective of this study was to evaluate the reliability of cloacal swabs to detect Campylobacter in young chicks. Day-old chicks (n = 25) were gavaged with 101–3 or 106 Campylobacter coli (C. coli) gentamicin-resistant marker strain. Batches of chicks were placed in separate isolation units, and 7, 10, 14, or 21 d post challenge, 10 birds per group were cloacally swabbed shallow (9 mm) and deep (24 mm). Swabs were placed into 5 mL of Tecra® broth, vortexed, and streaked for isolation onto Campy Cefex agar plus 200 ppm of gentamicin. After swabbing, birds were sacrificed and one cecum was quantitatively analyzed for C. coli from the control group; both ceca from all challenged birds were analyzed for C. coli. At 14 d post challenge, 95% of the shallow and 90% of the deep swabs were positive. Even with a low inoculum of 103, C. coli achieved a high degree of cecal colonization, and the cloacal swab (either shallow or deep) proved reliable for detecting C. coli. Birds challenged with >102, after 7 and 14 d were colonized with >106 cells. After 7 d, all shallow and deep swabs were positive for C. coli, regardless of challenge dose. Since it might not be practical in industry to process the swabs the d of collection, we looked at the reliability of cloacal swabs after freezing for up to 21 days. When the level in the ceca was high, recovery of C. coli was excellent, but when the level was low (± 102 inoculum level), recovery was very unreliable. If the levels of Campylobacter are relatively high (log ≥ 6.0) in the ceca, both the shallow and deep swabs, unfrozen or frozen, are reliable, nondestructive methods to detect this microorganism. [ABSTRACT FROM AUTHOR]

Campylobacter is considered an important foodborne pathogen, having a high impact on public health burden. Source allocation studies identified broiler meat as the most important food transmission vehicle of Campylobacter . In this regard, microbiological limits are under development in the EU and characterisation of microbial contamination seems to be an essential step towards implementing sampling schemes. The aim of this study was to analyse broiler batches processed at three conventional slaughterhouses in Germany for their Campylobacter load at the end of processing. Microbial data of positive and negative batches from the studied slaughterhouses were used to fit different statistical distributions (i.e., lognormal, Poisson-lognormal and negative binomial) by means of the maximum likelihood estimation method. Finally, the performance of microbiological criteria was assessed by calculating the probability of accepting batches using two microbiological limits recently discussed by the European Food Safety Authority (500 and 1000 cfu/g). Samples of broiler neck skin and caecal content (356 batches) were collected in three German slaughterhouses (A, B and C) for fresh meat production from conventionally reared broilers during the period from July 2013 to June 2016. Microbial analyses of five pooled neck skin samples and one pooled caecal sample per batch were performed. In this study, the prevalence of Campylobacter caecum positive broiler batches processed at slaughterhouses A, B and C was 42.7 (n = 117), 21.7 (n = 120) and 47.9% (n = 119), respectively. The Campylobacter mean count in neck skin samples of positive batches was highest in slaughterhouse A, with 2.9 ± 0.6 log 10 cfu/g compared to slaughterhouse B with 2.7 ± 0.7 log 10 cfu/g and slaughterhouse C with 2.7 ± 0.7 log 10 cfu/g. The results showed that distributions skewed to the right, thus indicating a high proportion of low microbial counts in the samples. Negative binomial regression provided a better fit at low contamination levels (<100 cfu/g), while the Poisson-lognormal distribution described both within- and between-batch variability better at higher microbial counts. The fitted statistical distributions were further used to evaluate the performance of different microbiological criteria based on an acceptance sampling approach in relation to the defective units based on different limits (mean counts) set at each slaughterhouse. Results obtained will help food business operators to evaluate the status of microbiological hygiene and safety in relation to Campylobacter contamination, also assisting them in setting their own limits of acceptance for process improvement under consideration of existing legal requirements. [ABSTRACT FROM AUTHOR]

In the last decades, a more prudent and rational use of antimicrobials has been progressively directed towards animal production to reduce antimicrobial selective pressure and antimicrobial resistance (AMR) in microorganisms and safeguard the antimicrobial efficacy of treatments in human medicine. This systematic review evaluated the effectiveness of interventions that have been applied to reduce or improve veterinary antimicrobial usage and aimed at decreasing resistant bacteria in chicken broiler and pig production contexts. Original articles were identified by searching PubMed™, Scopus™, The Cochrane Library™, and Web of Science™, and grey literature by searching DANS EASY™, WorldCat™ and RCAAP™. Inclusion criteria included: chicken broiler or pig populations (predestined for meat production), interventions intended to reduce/improve antimicrobial use, comparator with standard or no use of antimicrobials, outcomes related to prevalence of resistant bacteria, farm level studies, original data, and analytical observational studies. Data was extracted from eligible studies and meta-analysis using random or fixed effects models was conducted for combinations including type of intervention, bacterial species, production type and animal populations. Models were selected according to heterogeneity between studies. The effectiveness of interventions was assessed using pooled odds ratio of resistance to antimicrobial substances/classes by bacteria for associations between animal populations with and without intervention. A total of 46 studies were eligible for review. For chicken broilers, most interventions were identified as antimicrobial restrictions on all non-therapeutic use (46%), complete restriction (27%), and prohibition on antimicrobials used for growth promotion (23%). As for pig populations, restrictions were mainly observed on all non-therapeutic use (37%), complete restriction (37%) and group treatments (22%). For meta-analysis, 21 studies were pooled after assessment of existing combinations. These combinations demonstrated a protective effect for most antimicrobial classes in Escherichia coli , Campylobacter and Enterococcus isolates from samples of chicken broilers as well in Escherichia coli and Campylobacter spp. from samples of pigs, compared to animals raised under conventional production or without intervention. Increased odds of resistance were only observed for cephalosporins in E. coli and broilers raised without antimicrobials, and to fluoroquinolones and quinolones in Campylobacter and pigs raised without antimicrobials, compared to conventional production. Our study indicates that organic production, antimicrobial-free farms, and group treatment restrictions are recommended for AMR reduction, providing information that may support decision-making to tackle AMR and better reporting to improve comparability of results between studies. • Organic and antimicrobial-free farms had a protective effect on bacterial resistance to most assessed antimicrobials. • Fluoroquinolones- and quinolones-resistant Campylobacter may persist in farms that raise pigs without antimicrobials. • Restrictions on group treatments may help to decrease resistance in Escherichia coli from conventionally raised pigs. • Studies assessing effects of interventions need to ensure a better reporting to improve comparability between studies. [ABSTRACT FROM AUTHOR]

A methodology is presented to optimize a sampling plan for retail products to monitor the prevalence of foodborne pathogens in relation to disease burden. The optimization procedure links an exposure assessment, a quantitative measure for disease burden (i.e. Disability Adjusted Life Years), number of samples to be analyzed (based on uncertainty of the prevalence estimate) and costs for sample analysis. The methodology attributes DALY's on ‘pathogen-matrix level’ to ‘pathogen-food products’ using the exposure assessment. The subsequent procedure includes the number of samples that need to be analyzed per retail product such that the prevalences can be monitored within a preset uncertainty bound, and the costs per sample. The final optimization step sorts pathogen-product combinations using the costs per DALY criterion which results in a monitoring program with a maximum number of DALY's given a certain amount of money. An optimized sampling plan was established for four foodborne pathogens on meat products: Campylobacter in pork and poultry meat, Salmonella in pork, Toxoplasma in pork and Shiga-toxin producing Escherichia coli (STEC) O157 in beef, veal, and mutton/lamb. Results show that Campylobacter on poultry products and one Toxoplasma – pork combination consitute the top 10 in the proposed Dutch public health risk meat monitoring program. This optimized sampling plan monitors 98% of the total amount of DALY's attributed to the considered pathogen-animal species combinations. At the same time, the procedure gives insight in how the preset optimization criteria leads to the proposed set of pathogen-product combinations. An iterative implementation of updated model input (prevalences, DALY estimates) will lead to an up-to-date optimized risk based monitoring program. [ABSTRACT FROM AUTHOR]

Campylobacter can be difficult to recover from complex samples due to overgrowth by background bacteria. A 0.45- or 0.65-μm-pore-size filter overlaid on agar plates can be used as a means to separate Campylobacter from confounding non-Campylobacter cells, facilitating detection on solid plating media. It is unclear what percentage of cells in a Campylobacter suspension passes through a filter and results in visible colonies. The objective of this study was to compare the number of Campylobacter cells detected by the filter method with those detected by direct plating and determine if the filter method can be used to estimate cellular density of an unknown Campylobacter in suspension. Overnight liquid cultures of six subtypes of Campylobacter jejuni and six of Campylobacter coli, all originally detected in chicken samples, were used for this study. Motility of isolates was tested by using a stab into soft agar, incubating plates, and measuring colony size. Each subtype was applied to Campy-Cefex agar directly and through a 0.45- or 0.65-μm-pore-size filter. Filters were removed, plates were incubated, and colonies were counted; three replications were conducted. Mean recovery by direct plating was 8.3 log CFU/mL. Regardless of pore size, the overall mean number of Campylobacter detected by using the filter method was significantly less than that using direct plating (P < 0.05). The mean difference between direct plating and plating though a 0.65-μm-pore-size filter for motile Campylobacter was log 2.4 CFU/mL, with a 95% confidence interval of ±0.2 log CFU/mL. [ABSTRACT FROM AUTHOR]

Although campylobacteriosis caused by Campylobacter jejuni and Campylobacter coli has been increasingly reported worldwide owing to the consumption of contaminated poultry and fresh produce, the current detection protocols are not selective enough to inhibit unspecific microbes other than these pathogens. Five antibiotics were separately added to Bolton broth, and the survival rates of 18 Campylobacter spp. and 79 non-Campylobacter spp. were evaluated. The survival rate of the non-Campylobacter spp. was the lowest in Bolton broth with rifampin (6.3%), followed by cefsulodin (12.7%), novobiocin (16.5%), and potassium tellurite and sulfamethozaxole (both 17.7%). Also the most effective concentration of rifampin was found to be 12.5 mg/L, which markedly inhibited non-Campylobacter strains while not affecting the survival of Campylobacter strains. After the Campylobacter spp. were enriched in Bolton broth supplemented with 12.5 mg/L rifampin (R-Bolton broth), CampyFood Agar (CFA) was found to be better in selectively isolating the pathogens in the enrichment broth than the International Organization for Standardization method of using modified charcoal cefoperazone deoxycholate agar (mCCDA) for this step. When applied to natural food samples—here, romaine lettuce, pepper, cherry tomato, Korean leek, and chicken—the R-Bolton broth–CFA combination decreased the number of false-positive results by 50.0, 4.2, 20.8, 50.0, and 94.4%, respectively, compared with the International Organization for Standardization method (Bolton broth–mCCDA combination). These results demonstrate that the combination of R-Bolton broth and CFA is more efficient in detecting C. jejuni and C. coli in poultry and fresh produce and thus should replace the Bolton broth–mCCDA combination. [ABSTRACT FROM AUTHOR]

Campylobacter is a leading cause of foodborne diarrheal illness worldwide, and the emergence of antimicrobial-resistant strains is a major global public health concern. The goal of this study was to compare the activity of different fluoroquinolone antibiotics against ciprofloxacin-resistant Campylobacter jejuni and Campylobacter coli. Isolates from retail meats collected between 2002 and 2009 were selected based on their in vitro susceptibility testing results against ciprofloxacin. In total, 289 C. jejuni and 165 C. coli were collected and analyzed. All ciprofloxacin-resistant isolates had a single mutation (Thr86Ile) in their gyrase A (gyrA) gene and exhibited decreased susceptibility to all eight fluoroquinolones tested. Gatifloxacin, enrofloxacin, and levofloxacin showed greater activity than the other fluoroquinolone drugs in both ciprofloxacin-sensitive and -resistant strains. [ABSTRACT FROM AUTHOR]

The safety of ready-to-eat products such as cured pork loins must be guaranteed by the food industry. In the present study, the efficacy of the dry curing process of pork loins obtained from free-range pigs in the reduction of three of the most important foodborne pathogens is analysed. A total of 28 pork loin segments, with an average weight of 0.57 ± 0.12 kg, were divided into four groups with three being inoculated by immersion with 7 log CFU/ml of either Salmonella Typhimurium, Campylobacter coli or Listeria innocua and the last one inoculated by immersion with sterile medium (control group). The loin segments were treated with a seasoning mixture of curing agents and spices, packed in a synthetic sausage casing and cured for 64 days. Microbiological analysis, pH and water activity (a w ) were assessed at four stages. The values of pH and a w decreased with curing time as expected. S. Typhimurium and C. coli dropped significantly (3.28 and 2.14 log units, respectively), but limited reduction of L. innocua (0.84 log unit) was observed along the curing process. In our study, three factors were considered critical: the initial concentration of the bacteria, the progressive reduction of pH and the reduction of a w values. Our results encourage performing periodic analysis at different stages of the manufacturing of dry cured pork loins to ensure the absence of the three evaluated foodborne pathogens. [ABSTRACT FROM AUTHOR]

Thermophilic Campylobacter spp. are one of the primary causes of bacterial human diarrhea. The consumption of poultry meats, by-products, or both is suspected to be a major cause of human campylobacteriosis. The aims of this study were to determine the prevalence of thermophilic Campylobacter spp. in fresh poultry meat and poultry by-products by conventional culture methods and to confirm Campylobacter jejuni and Campylobacter coli isolates by using the multiplex PCR assay. Two hundred fifty fresh poultry samples were collected from a variety of supermarkets and slaughterhouses located in Sfax, Tunisia, including chicken (n =149) and turkey (n =101). The samples were analyzed using conventional microbiological examinations according to the 2006 International Organization for Standardization method (ISO 10272-1) for Campylobacter spp. Concurrently, a real-time PCR was used for identification of C. jejuni and C. coli. Of the 250 samples of poultry meat and poultry by-products, 25.6% (n = 64) were contaminated with Campylobacter spp. The highest prevalence of Campylobacter spp. was found in chicken meat (26.8%) followed by turkey meat (23.7%). Among the different products, poultry breasts showed the highest contamination (36.6%) followed by poultry by-products (30%), poultry wings (28%) and poultry legs (26%) showed the lowest contamination, and no contamination was found on neck skin. Of the 64 thermophilic Campylobacter isolates, C. jejuni (59.7%) was the most frequently isolated species and 10.9% of the isolates were identified as C. coli. All of the 64 Campylobacter isolates identified by the conventional culture methods were further confirmed by PCR. The seasonal peak of Campylobacter spp. contamination was in the warm seasons (spring and summer). The study concluded that high proportions of poultry meat and poultry by-products marketed in Tunisia are contaminated by Campylobacter spp. Furthermore, to ensure food safety, poultry meats must be properly cooked before consuming. [ABSTRACT FROM AUTHOR]

Irrigation water has been identified as a possible source of vegetable contamination by foodborne pathogens. Risk management for pathogens such as Campylobacter spp. and Listeria monocytogenes in fields can be influenced by the source of the irrigation water and the time interval between last irrigation and harvest. Plots of romaine lettuce were irrigated with manure-contaminated water or aerated pond water 21, 7, or 3 days prior to harvesting, and water and muck soil samples were collected at each irrigation treatment. Lettuce samples were collected at the end of the trials. The samples were tested for the presence of Campylobacter spp. and L. monocytogenes. Campylobacter coli was isolated from 33% of hog manure samples (n = 9) and from 11% of the contaminated water samples (n = 27), but no lettuce samples were positive (n = 288). L. monocytogenes was not found in manure, and only one sample of manure-contaminated irrigation water (n = 27) and one lettuce sample (n = 288) were positive. No Campylobacter or L. monocytogenes was recovered from the soil samples (n = 288). Because of the low incidence of pathogens, it was not possible to link the contamination of either soil or lettuce with the type of irrigation water. Nevertheless, experimental field trials mimicking real conditions provide new insights into the survival of two significant foodborne pathogens on romaine lettuce. [ABSTRACT FROM AUTHOR]

Ground poultry products are frequently contaminated with foodborne pathogens. With the potential for increased regulatory scrutiny, it is important to use sufficient intervention strategies to control pathogen levels effectively. A large proportion of the bacteria introduced to ground chicken are likely to come from broiler skin, which is added to achieve target fat content and maintain product texture and taste. In this research, antimicrobials, including 50 ppm of chlorine and 1,200 ppm of peracetic acid (PAA), were applied in a postchill system to reduce the number of Salmonella Typhimurium, Listeria monocytogenes , and Campylobacter coli inoculated on chicken skin used to formulate ground chicken. Results showed that chlorine provided no significant effect in reducing the number of pathogens in ground chicken made with treated skin compared with water treatment but that it did help decrease pathogens in postchill water. PAA was found to be an effective (P ≤ 0.05) antimicrobial agent, not only in reducing the number of pathogens on ground chicken, but also in postchill water. Treating chicken skin with PAA prior to inclusion in ground chicken can be an effective intervention strategy to lessen contamination in a ground chicken meat product. [ABSTRACT FROM AUTHOR]

Chicken products are known as main vehicles for human infections and foodborne illness with campylobacteriosis . This study describes a rapid and sensitive method for the detection of C. coli and C. jejuni from eggs, chicken meat, and chicken broth using the multiplex real-time recombinase polymerase amplification (Rti-RPA) and its comparison to a culture dependent method. The optimal procedure for the multiplex Rti-RPA assay was set in isothermal condition at 45 °C for 20 min and the detection sensitivity was at 1 CFU/reaction in pure culture. In the case of food applications, the detection limit of C. coli and C. jejuni using the RPA assay was 1 CFU/mL from chicken broth, 10 3 CFU/g from egg and chicken meat samples without a pre-enrichment procedure, and 1 CFU/g from 24 h enriched egg and chicken meat samples. Quantifications of Campylobacter spp. in spiked food samples by the RPA assay were not significantly different with the results by the culture method. This multiplex Rti-RPA assay is considerably faster than other DNA amplification methods and exhibits no losses in detection sensitivity and specificity. This multiplex Rti-RPA is a simple, rapid detection and quantification assay that is completed within a 20 min runtime. The assay is applicable in the surveillance of C. coli and C. jejuni contamination in eggs and chicken products. [ABSTRACT FROM AUTHOR]

Campylobacter species are known to be able to produce biofilm, which represents an ideal protective environment for the maintenance of such fragile bacteria. Since the genetic mechanisms promoting biofilm formation are still poorly understood, in this study we assessed the ability of C. jejuni (n = 7) and C. coli (n = 3) strains isolated from diseased poultry, and previously characterized by whole genome sequencing, to form biofilm. The in vitro analyses were carried out by using a microtiter based protocol including biofilm culturing and fixation, staining with crystal violet, and measurement of the optical density (OD 570). The ability to form biofilm was categorized into four classes (no, weak, moderate, and strong producers). Potential correlations between OD 570 and the presence/absence of virulence determinants were examined. The C. jejuni were classified as no (n = 3), weak (n = 2), and moderate (n = 2) biofilm producers; however, all possessed genes involved in chemotaxis, adhesion, and invasion to the host cells. No genes present exclusively in biofilm producers or in non-biofilm producers were identified. All C. coli were classified as weak producers and showed a similar set of virulence genes between each other. A trend of increased mean OD 570 was observed in the presence of flaA and maf7 genes. No association between biofilm production classes and the explanatory variables considered was observed. The results of this study suggest that further investigations are needed to better identify and characterize the genetic determinants involved in extra-intestinal Campylobacter biofilm formation. • Extra-intestinal Campylobacter jejuni and C. coli are able to produce biofilm. • Motility and adherence factors may be involved in Campylobacter biofilm formation. • flaA and maf7 genes may contribute to biofilm production in Campylobacter. • Biofilm-producing Campylobacter spp. may be a threat for public health. [ABSTRACT FROM AUTHOR]

Campylobacteriosis is one of the most common zoonotic diseases in North America. As opposed to humans, animal infections caused by Campylobacter spp. are often asymptomatic. In this study, data collected through the Canadian Dairy Network for Antimicrobial Stewardship surveillance system were used to determine the proportion of Campylobacter spp. and antimicrobial resistant isolates recovered from dairy cattle herds. Additionally, the association of antimicrobial use (AMU) with fecal carriage and antimicrobial resistance (AMR) of Campylobacter spp. were investigated. Pooled fecal samples from 5 animals from each production phase (pre-weaned calves, post-weaned heifers, lactating cows), and a manure storage sample were collected from 140 dairy herds across Canada. Samples were cultured using selective media, and Campylobacter isolates were speciated using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibilities were determined using the minimum inhibitory concentration test, and interpretation was made according to the Clinical and Laboratory Standards Institute. Two multilevel logistic regression models were used to investigate the association between the AMU with the isolation and antimicrobial resistance in Campylobacter spp. Of 560 samples, 63.8% were positive for Campylobacter spp., and 96% of the participating farms had at least one sample source (i.e., calves, heifers, lactating cows, or manure storage) positive for Campylobacter spp. Overall, 54.3% of the Campylobacter spp. isolates were resistant to at least one antimicrobial. Resistance to tetracycline was observed in 49.7% of the Campylobacter spp. isolates, followed by ciprofloxacin (19.9%) and nalidixic acid (19.3%). The proportion of multi-drug resistant (≥3 antimicrobial classes) Campylobacter spp. isolates was low (0.3%); however, 15.6% were resistant to two different classes of antimicrobials. Samples collected from lactating cows, heifers, and manure storage were more likely to be positive for Campylobacter spp. compared to calves. Total AMU was associated with a decreased probability of recovering Campylobacter spp. In addition, AMR to either tetracycline or ciprofloxacin had an interaction with antimicrobial use. The probability of resistance to tetracycline increased for each unit increase in the total AMU (Defined Course Dose/100 animal-years), while the probability of resistance to ciprofloxacin decreased. Campylobacter coli isolates were more likely to be resistant to ciprofloxacin and tetracycline when compared to C. jejuni. Our study demonstrated that Campylobacter spp. is widespread among Canadian dairy farms, and a higher proportion of resistance to tetracycline was identified. The total AMU was associated with increased resistance to tetracycline in Campylobacter spp. isolates; however, for ciprofloxacin the AMU was associated with decreased resistance. • The total antimicrobial (AMU) use was associated with increased resistance to tetracycline in Campylobacter spp. isolates. • The total AMU was associated with a decreased probability to recover a Campylobacter spp. from fecal and manure samples. • Fecal samples from pre-weaned calves were associated with a decreased probability of recovering a Campylobacter spp. isolate. [ABSTRACT FROM AUTHOR]

Salmonella and Campylobacter spp. are important foodborne pathogens and increased knowledge on the potential for egg transmission is beneficial when developing interventions. Horizontal and vertical transmission of Salmonella into poultry flocks is well documented to occur, while only horizontal transmission of Campylobacter has been confirmed. The objective of this study was to develop and verify injection inoculation methodology for hatching eggs that could be used for Salmonella and Campylobacter to simulate vertical transmission, in ovo. Six experiments were performed: Experiments 1 through 3 established inoculum levels of Salmonella Enteritidis (103 CFU) or Campylobacter coli (102 CFU) and Experiments 4 through 6 compared recovery from inoculated eggs that were incubated or cold stored (5 d). Eggs were inoculated via injection into the albumen or yolk and egg contents and viable embryos were sampled. Results from Experiments 1 through 3 indicated that incubation directly after injection resulted in higher embryonic mortality for yolk (90%) compared to albumen (35%) injection, though there was at least 1 positive embryo for each injection site at D5 and D15. For Experiment 4, eggs were albumen injected with Salmonella and cold-storage versus incubation resulted in few significant differences in recovery. For Experiments 5 and 6, Campylobacter injected eggs that were cold-stored had 100% enriched recovery from egg contents of albumen injection eggs compared to 0 to 40% for incubated eggs. This study establishes injection inoculation as a valid methodology for simulation and evaluation of the potential for Salmonella and Campylobacter to be vertically transmitted in ovo. [ABSTRACT FROM AUTHOR]

Campylobacter coli is the second most incriminated species of Campylobacter in cases of human campylobacteriosis, often isolated from chicken meat. Brazil is the largest exporter of chicken meat in the world, which makes the characterization of Brazilian isolates crucial for the establishment of control measures. Eighty-three C. coli strains isolated from chicken carcasses in three states in Brazil were tested by conventional PCR for 14 virulence genes classified into five categories: biofilm formation (flA , luxS), secretion system (ctdABC , hcp), invasion and colonization (cadF , ciaB , pldA), stress adaptation (cbrA , htrA , dnaJ), and Guillain-Barré syndrome (neuA , cstII). A broth microdilution test was performed to test sensitivity to ciprofloxacin and erythromycin. The proteomic similarity between the isolates was established through a dendrogram using data obtained from MALDI-TOF. The gene profile showed high potential for invasion and colonization. The strains showed 89.2% resistance to ciprofloxacin and 55.4% to erythromycin. Brazilian state I showed widespread genetic diversity, states II and III showed local specificity. Our results suggest high virulence and resistance, evidence of cross contamination and maintenance of virulent genotypes, emphasizing the need for control measures in Brazilian slaughterhouses. • Campylobacter coli showed a great potential for invasion and colonization. • The strains were highly resistant to ciprofloxacin and enrofloxacin. • State I showed high genetic diversity, states II and III showed local specificity. [ABSTRACT FROM AUTHOR]

• qPCR recovery efficiency of Campylobacter spp. from wastewater varies with concentrations. • Adsorption to sewer biofilms leads to large reduction of C. jejuni and C. coli in wastewater phase. • Gravity sewer, not rising main, conditions enhanced the total decay of C. jejuni and C. coli. • In-sewer decay induce significant variance of the WBE back-estimation of C. jejuni and C. coli. Campylobacter jejuni and coli are two main pathogenic species inducing diarrhoeal diseases in humans, which are responsible for the loss of 33 million lives each year. Current Campylobacter infections are mainly monitored by clinical surveillance which is often limited to individuals seeking treatment, resulting in under-reporting of disease prevalence and untimely indicators of community outbreaks. Wastewater-based epidemiology (WBE) has been developed and employed for the wastewater surveillance of pathogenic viruses and bacteria. Monitoring the temporal changes of pathogen concentration in wastewater allows the early detection of disease outbreaks in a community. However, studies investigating the WBE back-estimation of Campylobacter spp. are rare. Essential factors including the analytical recovery efficiency, the decay rate, the effect of in-sewer transport, and the correlation between the wastewater concentration and the infections in communities are lacking to support wastewater surveillance. This study carried out experiments to investigate the recovery of Campylobacter jejuni and coli from wastewater and the decay under different simulated sewer reactor conditions. It was found that the recovery of Campylobacter spp. from wastewater varied with their concentrations in wastewater and depended on the detection limit of quantification methods. The concentration reduction of Campylobacter. jejuni and coli in sewers followed a two-phase reduction model, and the faster concentration reduction during the first phase is mainly due to their partitioning onto sewer biofilms. The total decay of Campylobacter. jejuni and coli varied in different types of sewer reactors, i.e. rising main vs. gravity sewer. In addition, the sensitivity analysis for WBE back-estimation of Campylobacter suggested that the first-phase decay rate constant (k 1) and the turning time point (t 1) are determining factors and their impacts increased with the hydraulic retention time of wastewater. [Display omitted] [ABSTRACT FROM AUTHOR]

Lincomycin is widely used in respiratory and gastrointestinal infection in veterinary medicine and food animal production. Campylobacter members are vital foodborne pathogens causing campylobacteriosis, and the resistance to lincosamides is seldom reported. To date, only the rRNA methyltransferase Erm(B) has been confirmed to be associated with lincosamides resistance in Campylobacter. In this study, we identified a lnu (C) variant conferring lincomycin resistance in this pathogen of chicken origin. The Lnu(C) encoded by this gene variant showed substitution at position 8 (Asn8Lys), 11 (Phe11Leu) and 112 (Leu112Phe), when compared with the firstly reported Lnu(C) from Streptococcus agalactiae. Cloning of the lnu (C) variant into lincosamide-susceptible Campylobacter jejuni NCTC 11168 confirmed its function in conferring resistance to lincomycin with the 32-fold increased MICs. Sequencing analysis showed that the lnu (C) variant was located within a MTn Sag1 -like transposon together with insLNU , which is inserted between panB and cj0299 genes on the chromosome. lnu (C) gene was distributed among C. coli globally, and various STs were involved in the dissemination of lnu (C). Although transposition mediated by MTn Sag1 -like transposon failed to occur, the horizontal transfer mediated by natural transformation and reservoir for resistance genes may facilitate their adaptation to the antimicrobial selection pressure in chickens, which should not be ignored. • A lnu (C) variant was identified in Campylobacter coli from chicken origin. • The lnu (C) variant could confer lincomycin resistance. • lnu (C) was distributed among C. coli globally and various STs were involved. • Homologous recombination is the major mechanism for lnu (C) transformation. [ABSTRACT FROM AUTHOR]

A total of 651 samples from broiler cecal samples, carcasses, carcass parts collected at the slaughterhouse level in Sichuan Province of China were examined for the occurrence of Campylobacter jejuni and Campylobacter coli . After confirmed by species-specific multiplex PCR, the recovered isolates were examined for resistance to antimicrobials using an agar dilution method and investigated for the mutation of gyrA , tetO gene and V domain of 23S rRNA as well as the presence of class 1 integron and the associated gene cassettes. In addition, the genotype relatedness of the isolates was determined by pulsed-field gel electrophoresis (PFGE) profiling. The prevalence of Campylobacter was 56.1% in cecal samples, 31.0% in carcasses and 17.0% in carcass parts, respectively. Among them, C . jejuni accounted for 24.6% and C . coli occupied 20.0% of the samples. The strains of C . jejuni were most frequently resistant to ciprofloxacin (88.1%), followed by resistance to tetracycline (79.4%) and levofloxacin (78.1%). Most of the C . coli isolates were resistant to ciprofloxacin (100%), tetracycline (98.5%), levofloxacin (98.5%), clindamycin (98.5%) and erythromycin (93.9%). Antimicrobial resistance profiling showed that 93.7% of campylobacters were multidrug resistant (MDR) strains. Moreover, class 1 integrons were detected in 98.6% of MDR campylobacters, among which 98.7% were positive for C . jejuni and 98.5% for C . coli . Three kinds of gene cassettes-associated amplicons were identified and the amplicons profile of 1000–750–500–250 bp was the predominant pattern linked to the aminoglycoside resistance gene of aadA 2. The presence of mutation in gyrA , tetO and 23S rRNA between C . jejuni and C . coli varied from 89.7% to 97.3%, 96.6% to 94.1%, and 95.0% to 96.7%, respectively. Finally, the results of PFGE indicated that, 33 PFGE profiles were generated among 78 isolates of C . jejuni and the similarity level ranged from 42.1% to 99.1%. By contrast, 15 PFGE patterns were produced among 68 C . coli isolates sharing a similarity level of 54.0%–99.7%. Comparison of the PFGE and antimicrobial resistance profiles of the isolates reflected the high genetic diversity of Campylobacter tested. A poor correlation among the antimicrobial resistance patterns, resistance determinants and PFGE genotypes was observed in C . jejuni . Our study showed that there were several points of cross-contamination during broiler slaughter, and a high diversity of PFGE types in the Campylobacter isolates with high resistances to ciprofloxacin and tetracycline. [ABSTRACT FROM AUTHOR]

Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contamination with Campylobacter is largely unknown. In the Netherlands, the massive poultry culling to control the 2003 avian influenza epidemic coincided with a 44–50% reduction in human campylobacteriosis cases in the culling areas, suggesting substantial environment-mediated spread of poultry-borne Campylobacter . We inferred the origin of surface water Campylobacter jejuni and Campylobacter coli strains in Luxembourg and the Netherlands, as defined by multilocus sequence typing, by comparison to strains from poultry, pigs, ruminants, and wild birds, using the asymmetric island model for source attribution. Most Luxembourgish water strains were attributed to wild birds (61.0%), followed by poultry (18.8%), ruminants (15.9%), and pigs (4.3%); whereas the Dutch water strains were mainly attributed to poultry (51.7%), wild birds (37.3%), ruminants (9.8%), and pigs (1.2%). Attributions varied over seasons and surface water types, and geographical variation in the relative contribution of poultry correlated with the magnitude of poultry production at either the national or provincial level, suggesting that environmental dissemination of Campylobacter from poultry farms and slaughterhouses can be substantial in poultry-rich regions. [ABSTRACT FROM AUTHOR]

Campylobacteriosis has become the leading bacterial zoonosis in humans in the European Union and other developed countries. There are many sources of human Campylobacter infections, but broilers and broiler meat have been shown to be the most important. In order to implement effective interventions that reduce the probability of Campylobacter colonisation of broiler flocks, it is essential to fully understand the risk factors involved. We present a bi-national risk factor survey comprising Campylobacter data from more than 5200 Danish and Norwegian indoor, conventional broiler flocks and the responses to a standardised questionnaire, with more than 40 explanatory variables from 277 Danish and Norwegian farms. We explored several models by using different combinations of the Danish and Norwegian data, including models with single-country datasets. All models were analysed using a generalized linear model using backwards elimination and forward selection. The results show that Norwegian broiler flocks had a lower risk of being colonised than Danish flocks. Farm specific variables that increased the risk of flocks becoming colonised with Campylobacter in both countries were: broiler houses older than five years; longer downtime (no. of days between flocks), probably a consequence of longer downtimes being associated with less focus on maintaining a high biosecurity level; broiler houses without a separate ante-room or barrier; and the use of the drinker nipples with cups or bells compared with nipples without cups. Additional country specific risk factors were also identified. For Norway, the risk of colonisation increased with increasing numbers of houses on a farm and when the water used for the broilers originated from surface water or bore holes instead of mains. For Denmark, having boot dips or low stocking density increased the risk of a flock becoming Campylobacter positive. The different model approaches allowed us to explore the effect of having a large number of data available to identify the significant variables. To a large extent, the country specific models identified risk factors that were also found in the bi-national model. However, the bi-national model identified more risk factors than the country specific models. This indicated that combining the data sets from the two countries did not disrupt the results but was beneficial due to the greater strength achieved in the statistical analyses and the possibility of examining interactions terms with the variable Country. [ABSTRACT FROM AUTHOR]