Campylobacter Isolates Research Articles

P-124. Ongoing Infections with Extensively Drug-Resistant Campylobacter jejuni Linked to Pet Store Puppies

Abstract Background Campylobacter is the leading cause of enteric bacterial infections in the United States, causing an estimated 1.5 million illnesses annually. Antimicrobial therapy can reduce the duration of illness and limit complications; macrolides and fluoroquinolones are typically recommended as first- and second-line treatment, respectively. The Centers for Disease Control and Prevention (CDC) investigates emerging resistance in Campylobacter strains to identify potential sources and collaborate on approaches to reduce its spread. Here, we describe extensively drug-resistant (XDR) strains linked to pet store puppies. Methods State public health departments perform whole genome sequencing on Campylobacter isolates as resources allow and submit sequence data to CDC. CDC assesses isolate relatedness using core genome multilocus sequence typing (cgMLST) and screens isolate sequences for resistance determinants to predict antimicrobial resistance (older isolates sequenced retroactively are also screened). We considered isolates to be XDR if they showed predicted resistance to macrolides, fluoroquinolones, and ≥ 3 additional antibiotic classes. Persons with XDR isolates were interviewed about exposures when possible. Results We identified three strains of XDR C. jejuni; the oldest isolates were from 2007. Although these strains are not highly related phylogenetically ( > 200 allele differences between strains by cgMLST), cases share epidemiological and clinical characteristics. As of June 30, 2023, CDC received reports of 274 cases associated with these strains. Affected individuals had a median age of 36 years (interquartile range, 20–52); 66% were female, and 23% were hospitalized. Of those affected, 97% reported contact with a dog before becoming ill, among whom 78% reported contact with a puppy from a pet store. Conclusion Clinicians should be aware that strains of XDR Campylobacter are circulating in the United States and should consider XDR infection when patients fail to respond to routine treatment, particularly if they have recently had contact with pet store puppies. Carbapenem antibiotics may be used for severe infections that are resistant to recommended treatment agents. Disclosures All Authors: No reported disclosures

A One Health approach for the genomic characterization of antibiotic-resistant Campylobacter isolates using Nanopore whole-genome sequencing

In response to the growing threat posed by the spread of antimicrobial resistance in zoonotic Campylobacter, a One Health approach was used to examine the genomic diversity, phylogenomic relationships, and the distribution of genetic determinants of resistance (GDR) in C. jejuni and C. coli isolates from humans, animals (ruminants, swine, and chickens), and avian food products collected during a regionally (Basque Country, Spain) and temporally (mostly 2021–2022) restricted sampling. Eighty-three C. jejuni and seventy-one C. coli isolates, most exhibiting resistance to ciprofloxacin and/or erythromycin, were whole-genome sequenced using Oxford Nanopore Technologies long-fragment sequencing (ONT). Multilocus sequence typing (MLST) analysis identified a high genomic diversity among isolates. Phylogenomic analysis showed that clustering based on the core genome was aligned with MLST profiles, regardless of the sample source. In contrast, accessory genome content sometimes discriminated isolates within the same STs and occasionally differentiated isolates from different sources. The majority of the identified GDRs were present in isolates from different sources, and a good correlation was observed between GDR distribution and phenotypic susceptibility profiles (based on minimum inhibitory concentrations interpreted according to the EUCAST epidemiological cutoff values). Genotypic resistance profiles were independent of genotypes, indicating no apparent association between resistance and phylogenetic origin. This study demonstrates that ONT sequencing is a powerful tool for molecular surveillance of bacterial pathogens in the One Health framework.

Sixty-five draft genome sequences of Campylobacter coli and Campylobacter jejuni from table egg layer chickens.

We present the draft genomic sequences of 65 isolates of Campylobacter spp. (38 Campylobacter coli and 27 Campylobacter jejuni) isolated from table egg-layer chickens, which are not generally associated with human transmission. Up to this time, there are no publicly available genomic sequences of Campylobacter isolated from laying hens.

Green Tea Extract (Theaceae; Camellia sinensis L.): A Promising Antimicrobial, Anti-Quorum Sensing and Antibiofilm Candidate Against Multidrug-Resistant Campylobacter Species.

Thermophilic Campylobacter species are among the main culprits behind bacterial gastroenteritis globally and have grown progressively resistant to clinically important antimicrobials. Many studies have been carried out to explore innovative and alternative strategies to control antibiotic-resistant campylobacters in animal reservoirs and human hosts; however, limited studies have been performed to develop efficient control schemes against Campylobacter biofilms. This study investigated the antimicrobial and antibiofilm activities of some herbal extracts against multidrug-resistant (MDR) Campylobacter species recovered from different sources using phenotypic and molecular techniques. The overall Campylobacter species prevalence was 21.5%, representing 15.25% and 6.25% for C. jejuni and C. coli, respectively. Regarding C. jejuni, the highest resistance rate was observed for amoxicillin-clavulanic acid and colistin (85.25% each), followed by cefotaxime (83.61%) and tetracycline (81.97%), whereas C. coli isolates showed absolute resistance to cefotaxime followed by erythromycin (92%) and colistin (88%). Remarkably, all Campylobacter isolates were MDR with elevated multiple antimicrobial resistance (MAR) indices (0.54-1). The antimicrobial potentials of green tea (Camellia sinensis), rosemary (Rosmarinus officinalis) and ginger (Zingiber officinale) extracts against MDR Campylobacter isolates were assessed by the disk diffusion assay and broth microdilution technique. Green tea extract showed a marked inhibitory effect against tested isolates, exhibiting growth inhibition zone diameters of 8 to 38 mm and a minimum inhibitory concentration (MIC) range of 1.56-3.12 mg/mL, unlike the rosemary and ginger extracts. Our findings reveal a respectable antibiofilm activity (>50% biofilm formation inhibition) of green tea against the preformed biofilms of Campylobacter isolates. Furthermore, real-time quantitative polymerase chain reaction (RT-qPCR) results showed a significant decrease (p < 0.05) in the expression levels of biofilm biosynthesis gene and its regulator (FlaA and LuxS, respectively) in Campylobacter isolates treated with the green tea extract in comparison with untreated ones. This is the first in vitro approach that has documented the inhibitory activity of green tea extract against MDR-biofilm-producing Campylobacter species isolated from different sources. Further in vivo studies in animals' models should be performed to provide evidence of concept for the implementation of this alternative candidate for the mitigation of MDR Campylobacter infections in the future.

Understanding antimicrobial resistance in Campylobacter isolates from poultry environments in Gujarat, India

Understanding antimicrobial resistance in Campylobacter isolates from poultry environments in Gujarat, India

Urban yellow-legged gull (Larus michahellis) and peri-urban Audouin's gull (Larus audouinii) as a source of Campylobacter and Salmonella of public health relevance.

Campylobacter spp. and Salmonella spp. are the leading cause of human enteric infections in the European Union. Some gull species act as reservoirs and play an important role in the epidemiology of these zoonotic agents. To gain insight into Campylobacter and Salmonella epidemiology we studied colonies of Audouin's gull (Larus audouinii) and yellow-legged gull (Larus michahellis) in Barcelona metropolitan area, Catalonia (north-eastern Spain). We assessed the occurrence, genetic diversity, virulence potential, and antimicrobial susceptibility of Campylobacter and Salmonella isolates recovered from gull faeces in different time periods within 2009-2018. The occurrence of Campylobacter was higher compared to Salmonella in both gull species. Also, the occurrence of both pathogens was significantly higher in Audouin's gull (45% for Campylobacter, 20% for Salmonella), than in yellow-legged gull (13% and 7%, respectively). All but one individual carried C. jejuni; the remaining positive yellow-legged gull carried C. lari. Salmonella serovar Typhimurium (including its monophasic variant) was the most frequent in both hosts followed by ser. Bredeney. Other serovars frequently associated with human salmonellosis (Infantis, London, Virchow) were only isolated from yellow-legged gulls. Multilocus Sequence Typing analyses showed that yellow-legged gull and not Audouin's gull carried several Campylobacter genotypes associated with human enteritis. Campylobacter isolates from both gull species revealed a high virulence potential, as opposed to Salmonella isolates which showed a lower prevalence of virulence-associated genes, particularly in Audouin's gull. Overall, a moderate to high frequency of antimicrobial resistance (including multidrug resistance) was found in both pathogens from both gull species. Campylobacter and Salmonella from yellow-legged gull showed a higher frequency of isolates resistant to antimicrobials of relevance in human medicine. Overall, our results highlight the potential public health threat associated with these gull species, particularly yellow-legged gull, in densely human populated areas.

Sharing of cmeRABC alleles between C. coli and C. jejuni associated with extensive drug resistance in Campylobacter isolates from infants and poultry in the Peruvian Amazon.

Campylobacter is a serious health threat because of the rapid progressive evolution of antimicrobial resistance and efficient transmission from zoonotic as well as human sources. Resistance to fluoroquinolones and macrolides is particularly concerning as this compromises the two most effective oral antibiotic agents currently available for human campylobacteriosis. Here, we report on the prevalence and worldwide distribution of the operon cmeRABC, which encodes an efflux pump conferring high levels of combined resistance to fluoroquinolones and macrolides in Campylobacter strains isolated from poultry (n = 75) and children (n = 177). These mutations were found to be highly prevalent in isolates from poultry (62.7%) and children (29.4%) in Iquitos, Peru. We investigated the population structure of genes in the cmeRABC operon and identified a potential genetic bottleneck for the cmeA and cmeB genes. While most cmeB alleles segregate by species, alleles associated with high resistance to fluoroquinolones and macrolides were found in both Campylobacter jejuni and Campylobacter coli. We inferred that the likely ancestry of these alleles was from C. jejuni and was later acquired by C. coli through recombination. Publicly accessible global genomic data from 16,120 Campylobacter genomes identified these mutations in approximately 6% of C. jejuni and C. coli isolates globally, with higher prevalence in samples from poultry in many countries, including Peru. Our findings suggest that these extensively drug-resistant Campylobacter strains originated from C. jejuni in poultry.IMPORTANCEAntimicrobial resistance in Campylobacter is a growing public health concern, driven by the rapid evolution and zoonotic transmission of resistant strains. This study focuses on mutations in the cmeABC efflux pump, which confer high resistance to fluoroquinolones and macrolides, the two most effective oral antibiotics for human campylobacteriosis. By analyzing genomes from poultry and children in Iquitos, Peru, as well as global genomic data sets, we identified a significant prevalence of these resistance-associated mutations, particularly in poultry and children. Our findings suggest that these mutations originated in Campylobacter jejuni and spread to C. coli through recombination. Globally, these mutations are found in approximately 6% of isolates, with higher prevalence in poultry in multiple countries. This research underscores the critical role of genomic epidemiology in understanding the origins, evolution, and dissemination of antimicrobial resistance and highlights the need to address poultry as a reservoir for resistant Campylobacter.

Molecular typing and antimicrobial susceptibility profiles of Campylobacter jejuni and Campylobacter coli Isolates from Patients and raw meat in Huzhou, China, 2021-2022.

Campylobacter species are zoonotic pathogens, and are considered to be the major foodborne pathogen that causes outbreaks and sporadic gastrointestinal illnesses both in developed and developing countries. In this study, the molecular typing and antimicrobial susceptibility profiles of Campylobacter jejuni and Campylobacter coli isolates from patients and raw meat between 2021 and 2022 in Huzhou were analyzed by using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and antimicrobial susceptibility testing. From September 1, 2021 to December 31, 2022, a total of 342 fecal specimens from diarrheal patients at a sentinel hospital in Huzhou and 168 samples of raw meat products collected from farmers' markets and supermarkets, were subjected to Campylobacter isolation and identification. The agar dilution method was used to determine resistance of the Campylobacter isolates to eleven antibiotics. In addition, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were performed to compare their genetic relationships. 78 Campylobacter isolates were recovered, comprising 58 isolates (74.36%, 58/78) of Campylobacter jejuni (34 patient isolates and 16 food isolates) and 20 isolates (25.64%, 20/78) of Campylobacter coli (6 patient isolates and 14 food isolates). Campylobacter has emerged as a predominant foodborne pathogen in the local region, with detection rate reached 11.70% among 342 diarrhea samples. The Campylobacter isolation rate in 168 raw meat was 22.62% (38/168), all originating from poultry meat, with chicken been the major source of infection (86.84%, 33/38). Both PGFE type and MLST data confirmed that Campylobacter stains circulating in Huzhou are genetically diverse, with Campylobacter jejuni isolates being more diverse than Campylobacter Coli. PFGE typing revealed 45 band patterns among 54 Campylobacter jejuni strains and 17 band patterns among 19 Campylobacter Coli strains. 50 Campylobacter jejuni strains from different sources were classified into 37 ST types, showing a dispersed distribution and encompassing over 12 clonal complexes (CCs), with CC-21 being the most prevalent CC (22.00%, 11/50). The distribution of ST types in the 18 Campylobacter Coli strains was relatively concentrated, with 83.33% (15/18) of isolates belonging to the CC-828. In this study, 2 groups of Campylobacter jejuni strains (PFGE J2-ST464 and PFGE J9-ST-2328) originated from humans and chickens showed high genetic homologies by comparing PFGE and MLST results. Besides, some disagreement between PFGE and MLST was observed for certain ST, indicating a weak correlation between PFGE and MLST for certain Campylobacter strains. Most of the Campylobacter isolates were highly resistant to nalidixic-acid, ciprofloxacin and tetracycline. The multiple antibiotic resistance of Campylobacter Coli (89.47%) is higher than Campylobacter jejuni (29.63%). Campylobacter is an important foodborne pathogen in both diarrheal patients and raw meat products in Huzhou City, exhibiting multiple antibiotic resistance and high level of genetic diversity.

Genomic epidemiology and genetic characteristics of clinical Campylobacter species cocirculating in West Bengal, India, 2019, using whole genome analysis.

Campylobacter species are the most common pathogens responsible for foodborne gastroenteritis worldwide. India is a region with frequent diarrheal infections and a high level of Campylobacter infection incidence, but the detailed genomic information is limited. This study aimed to characterize 112 isolates of Campylobacter from diarrhea patients at two hospitals in Kolkata, West Bengal, by whole genome analysis. The Campylobacter isolates consisted of 90 C. jejuni, 20 C. coli, and 2 C. lari isolates. Multilocus sequence typing analysis revealed that the largest sequence type (ST) populations were ST-2131 in C. jejuni and ST-830 in C. coli and seven novel STs were found in C. jejuni and one in C. coli. Notably, ST-2131, which is rarely seen elsewhere, was positive for a sialylated LOS-related gene (wlaN +neuA + cstIII) associated with Guillain-Barré syndrome. Antibiotic resistance factors predicted from the genome sequence included blaOXA variants (58.9%), tet(O) (54.5%), tet(W) (0.9%), ant(6)-Ia (0.9%), mutation in GyrA (T86I, T86I+D90N, T86I+P104S, T86I+D90N+P104S) (79.5%), and mutation in 23S rRNA (A2075G) (12.5%). In addition to the high drug resistance of Campylobacter in Kolkata, Campylobacter pathogens were circulating that may be associated with Guillain-Barré syndrome. This study indicates the importance of genomic analysis in the surveillance of pathogens, which provides genomic information on genetic diversity, virulence mechanisms, and determinants of antimicrobial resistance.

Effect of feeding biochar, oat hulls, yeast fermentate, and organic acids on reduction of Campylobacter in free-range broilers from hatching to slaughter.

The Campylobacter prevalence in free-ranging broiler flocks is usually higher than in conventional flocks, and effective interventions for this production type are needed. This study aimed to investigate the on-farm Campylobacter-reducing effect of feeding three feed additives or a water additive to broilers from hatching to slaughter. Newly hatched Ranger Gold broilers (n = 140) were randomly placed into five cages (n = 28/cage) within a flock of 6,000 broilers. The broilers had access to outdoor areas after day 36. Three groups received one of the following feed additives: 2 % (w/w) Biochar Charcoal Feed Granules for Poultry, 0.125 % Original XPC™ (Saccharomyces cerevisiae fermentate), or 15 % oat hulls. One group received organic acids added to water (Selko®pH E 0.2 % (v/v)), and one group was fed standard feed pellets and water (control). Half of the broilers were sacrificed on day 36 and the rest on day 56 for enumeration and isolation of Campylobacter in cecal contents by culturing. Air samples were collected weekly, and water and boot sock samples were also collected on days 4, 36, and 59 and analyzed for Campylobacter. A selection of the obtained Campylobacter isolates (n = 43) was whole-genome-sequenced. For the three successive flocks tested, all broilers became Campylobacter-positive before access to the outdoors on day 36. Adding biochar in the feed resulted in 0.70 log10 CFU/g lower C. jejuni than the control group on day 59. The addition of organic acids in water resulted in 0.62 log10 CFU/g lower C. jejuni than the control group on day 36. There was a noticeable difference in dominant sequence types (ST) between day 36 (ST52 and ST475) and day 59 (ST45). STs were genetically similar across rotations, indicating transmission of isolates between successive flocks or from the outdoor environment. High variability was seen between calls for further testing to confirm the demonstrated effect.

Culture-Based Standard Methods for the Isolation of Campylobacter spp. in Food and Water.

Campylobacter spp. is a major source of global gastrointestinal infections. Their enteric infections are linked to the consumption of undercooked poultry products, contaminated milk and water, and the handling of wild animals and birds. The detection of Campylobacter spp. in water and food samples mainly depends on culture-based techniques. Public Health England (PHE), the U.S. Food and Drug Administration (FDA), and the International Standard Organization (ISO) have standardized Campylobacter spp. isolation and enumeration procedures for food and water samples, which involve the usage of selective agar media and enrichment broth. Different types of selective plating and enrichment media have been prepared for Campylobacter spp. detection and assessment during regular food surveillance and food poisoning. To date, culture media remains the standard option for microbiological food analysis and has been approved by the U.S. Environmental Protection Agency (US EPA), Food and Agriculture Organization (FAO), and World Health Organization (WHO). This review discusses the standard microbiological protocols for Campylobacter spp. isolation and enumeration in food and water and evaluates detection media (pre-enrichment, selective enrichment, and selective plating) for their rational applications. Moreover, it also elaborates on the advantages and disadvantages of recent chromogenic culture media in Campylobacter spp.-oriented food surveillance. This review also highlights the challenges of culture-based techniques, future developments, and alternative methods for Campylobacter spp. detection in food and water samples.

Mutation-based mechanism and evolution of the potent multidrug efflux pump RE-CmeABC in Campylobacter

The resistance-nodulation-cell division (RND) superfamily of multidrug efflux systems are important players in mediating antibiotic resistance in gram-negative pathogens. Campylobacter jejuni, a major enteric pathogen, utilizes an RND-type transporter system, CmeABC, as the primary mechanism for extrusion of various antibiotics. Recently, a functionally potent variant of CmeABC (named RE-CmeABC) emerged in clinical Campylobacter isolates, conferring enhanced resistance to multiple antibiotic classes. Despite the clinical importance of RE-CmeABC, the molecular mechanisms for its functional gain and its evolutionary trajectory remain unknown. Here, we demonstrated that amino acid substitutions in RE-CmeB (inner membrane transporter), but not in RE-CmeA (periplasmic protein) and RE-CmeC (outer membrane protein), in conjunction with a nucleotide mutation in the promoter region of the efflux operon, are responsible for the functional gain of the multidrug efflux system. We also showed that RE-cmeABC is emerging globally and distributed in genetically diverse C. jejuni strains, suggesting its possible spread by horizontal gene transfer. Notably, many of RE-cmeABC harboring isolates were associated with the human host including strains from large disease outbreaks, indicating the clinical relevance and significance of RE-CmeABC. Evolutionary analysis indicated that RE-cmeB likely originated from Campylobacter coli, but its expansion mainly occurred in C. jejuni, possibly driven by antibiotic selection pressure. Additionally, RE-cmeB, but not RE-cmeA and RE-cmeC, experienced a selective sweep and was progressing to be fixed during evolution. Together, these results identify a mutation-based mechanism for functional gain in RE-CmeABC and reveal the key role of RE-CmeB in facilitating Campylobacter adaptation to antibiotic selection.

Gene profile of virulence, antimicrobial resistance and action of enterocins in Campylobacter species isolated from broiler carcasses.

Campylobacteriosis is among the most reported zoonoses in the world, caused by species of Campylobacter, this disease is characterized by gastroenteritis in humans. The main species involved is Campylobacter jejuni, followed by Campylobacter coli. Contaminated chicken meat is often identified as an important source of infection related to human cases and 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. The objective was to evaluate the contamination of chilled and frozen carcasses sold in the Northeast of Brazil, by Campylobacter species, identify virulence genes, evaluate bacterial resistance to antibiotics and verify the antimicrobial action of the Crude Extract Containing Enterocins (CECE) produced by a probiotic strain of Enterococcus faecium. In this study, 12 chilled carcasses and 12 frozen carcasses were collected, sold retail in supermarkets. The following regions of the carcass were sampled: breast skin, wing skin, belly skin, neck skin, gizzard and liver. Samples of chicken carcasses were analyzed following ISO 10272-2 guidelines for the isolation of Campylobacter spp. The isolates were tested by PCR to identify genus, species C. jejuni, C. coli and C. lari and genes cdtA, cdtB, cdtC, sodB, dnaJ, cmeA, cmeB, cmeC. The assessment of susceptibility to antibiotics was carried out using the standard disk diffusion method and the antimicrobial activity of CECE was determined using the Minimum Inhibitory Concentration (MIC), the methodologies followed the recommendations and cutoff points according to EUCAST and CLSI. A total of 376 isolates of Campylobacter spp. were obtained, among these, 26 (7.0%) were positive for C. jejuni and no isolates were detected for C. coli and C. lari. The highest frequency of C. jejuni was obtained in chilled carcasses with 23 isolates (88.5%, p < 0.0001), in frozen carcasses three isolates were obtained (11.5%). The most frequency site of C. jejuni was the chest skin (7/27.0%), followed by skin of the wing (6/23.0%), skin of the cloaca (5/19.0%), gizzard (4/15.0%), skin of the neck (2/8.0%) and liver (2/8.0%), no significant differences were found between the sites sampled. The gene frequency was determined in: cdtA (3/11.5%), cdtB (3/11.5%), cdtC (5/19.0%), sodB (9/34.5%), dnaJ (3/11.5%), cmeA (4/15.0%), cmeB (4/15.0%) and cmeC (4/15.0%). The three efflux pump genes were amplified in four isolates (15.3%) and all tested genes were amplified in three isolates (11.5%). All C. jejuni isolates (26/100.0%) were found to be multiresistant to three or more classes of antimicrobials. The index of multiple resistance to antimicrobial drugs (IRMA) ranged from 0.4 to 1.0 among isolates of C. jejuni. The antimicrobial activity of CECE was able to inhibit at least 98.5% of the growth of all C. jejuni isolates. Therefore, chilled chicken carcasses present a greater risk of contamination than frozen carcasses, for this reason it is necessary to adopt practices that avoid cross-contamination during the preparation of chicken meat, in order to prevent campylobacteriosis. Furthermore, the presence of multiresistant and potentially virulent isolates highlights the need for further investigations to better understand the use of enterocins as alternative methods in the control of Campylobacter.

Virulence and resistance genes in &lt;i&gt;Campylobacter spp&lt;/i&gt; isolated from asymptomatic children at Kibera slum in Nairobi, Kenya

BACKGROUND Campylobacter spp. are among the global causes of food poisoning and harbour virulence and resistance genes which play a role in invasion and antibiotic resistance. This study aimed to detect selected resistance and virulence genes in archived Campylobacter spp. isolated from children in Kibera, Nairobi, Kenya. MATERIALS AND METHODS This was a retrospective laboratory-based cross-sectional study on archived samples of Campylobacter spp. at the Department of Public Health, Pharmacology and Toxicology, University of Nairobi. Campylobacter isolates were cultured on modified charcoal cefoperazone deoxycholate agar (mCCDA) media. The presence of selected virulence genes and resistance genes was detected by multiplex polymerase chain reaction (mPCR). STATA13 was used in data analysis. Binary logistic regression was used to ascertain the association between the genes and the Campylobacter spp. RESULTS The Campylobacter spp. under this study were Campylobacter jejuni (C. jejuni) (50%), Campylobacter coli (C. coli) (38%) and Campylobacter lari (C. lari) (12%). The virulence genes detected were cgtB 37 (25%), pldA (20%), cdtA (17%), cdtB (16%), cdtC (6%), wlaN (5%), iam (4%), ciaB (4%) and VirB11 (3%). The virulence gene waaC was not detected in any of the isolates. The resistance genes detected were TetA (42%), Tet B (26%), GyrB (18%) and GyrA (14%). CONCLUSION The study revealed the presence of various virulence and resistance genes in Campylobacter spp. isolated from children. This information could help guide the development and enhancement of guidelines for the management of Campylobacter infections.

Whole-genome comparative analysis of the genetic, virulence and antimicrobial resistance diversity of Campylobacter spp. from Spain

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 blaOXA-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.

Campylobacter Diversity Along the Farm-to-Fork Continuum of Pastured Poultry Flocks in the Southeastern United States.

Consumer demand for pasture raised, antibiotic-free poultry products has led to an increase in pastured poultry operations within the United States. Given the level of environmental interaction and the potential increase in exposure to foodborne pathogens in these settings, a greater understanding of the prevalence and diversity of Campylobacter populations inherent within pastured poultry flocks is needed. To achieve this, 40 pastured poultry flocks from nine farms were sampled using a farm-to-fork strategy, and Campylobacter was isolated and characterised from preharvest (faeces, soil) through postharvest (caeca, whole carcass rinse) to the final product the consumer would purchase (whole carcass rinse). Campylobacter was isolated from 872 of 1820 samples, showing an overall prevalence of 47.91%. The caeca showed the highest (p < 0.05) Campylobacter load (4.64 log10 CFU/mL) and prevalence (95.5%), while the final product whole carcass rinses had the lowest (p < 0.05) Campylobacter load (0.32 log10 CFU/mL) and prevalence (15.45%), suggesting that the Campylobacter load in the caeca may not be indicative of the Campylobacter load on the final product. Of the 872 positive samples, 337 Campylobacter isolates were selected for further characterisation. Campylobacter jejuni and Campylobacter coli comprised 74.18% (250/337) and 21.95% (74/337) of the selected isolates respectively. While the Campylobacter isolates displayed resistance to several antibiotics, the most common resistance for both C. jejuni and C. coli was against tetracycline (55.86% and 70.31% respectively). Multidrug resistance phenotypes (≥ 3 antibiotic classes) were relatively low for both C. jejuni (2.80%) and C. coli (9.45%). Campylobacter load, prevalence and diversity were more affected by farm location than by the type of sample from which the Campylobacter was isolated. Overall, these results indicated a need for farm-specific Campylobacter mitigation strategies to ensure the safety of these increasingly in-demand poultry products.

Campylobacteriosis Outbreak Linked to Municipal Water, Nebraska, USA, 20211.

In September 2021, eight campylobacteriosis cases were identified in a town in Nebraska, USA. We assessed potential exposures for a case-control analysis. We conducted whole-genome sequencing on Campylobacter isolates from patients' stool specimens. We collected large-volume dead-end ultrafiltration water samples for Campylobacter and microbial source tracking testing at the Centers for Disease Control and Prevention. We identified 64 cases in 2 waves of illnesses. Untreated municipal tap water consumption was strongly associated with illness (wave 1 odds ratio 15.36; wave 2 odds ratio 16.11). Whole-genome sequencing of 12 isolates identified 2 distinct Campylobacter jejuni subtypes (1 subtype/wave). The town began water chlorination, after which water testing detected coliforms. One dead-end ultrafiltration sample yielded nonculturable Campylobacter and avian-specific fecal rRNA genomic material. Our investigation implicated contaminated, untreated, municipal water as the source. Results of microbial source tracking supported mitigation with continued water chlorination. No further campylobacteriosis cases attributable to water were reported.

A Decade of Antimicrobial Resistance in Human and Animal Campylobacter spp. Isolates.

Objectives: Campylobacter spp. remain a leading cause of bacterial gastroenteritis worldwide, with resistance to antibiotics posing significant challenges to treatment and public health. This study examines profiles in antimicrobial resistance (AMR) for Campylobacter isolates from human and animal sources over the past decade. Methods: We conducted a comprehensive review of resistance data from studies spanning ten years, analyzing profiles in resistance to key antibiotics, ciprofloxacin (CIP), tetracycline (TET), erythromycin (ERY), chloramphenicol (CHL), and gentamicin (GEN). Data were collated from various regions to assess global and regional patterns of resistance. Results: The analysis reveals a concerning trend of increasing resistance patterns, particularly to CIP and TET, across multiple regions. While resistance to CHL and GEN remains relatively low, the high prevalence of CIP resistance has significantly compromised treatment options for campylobacteriosis. Discrepancies in resistance patterns were observed between human and animal isolates, with variations across different continents and countries. Notably, resistance to ERY and CHL showed regional variability, reflecting potential differences in antimicrobial usage and management practices. Conclusions: The findings underscore the ongoing challenge of AMR in Campylobacter, highlighting the need for continued surveillance and research. The rising resistance prevalence, coupled with discrepancies in resistance patterns between human and animal isolates, emphasize the importance of a One Health approach to address AMR. Enhanced monitoring, novel treatment strategies, and global cooperation are crucial for mitigating the impact of resistance and ensuring the effective management of Campylobacter-related infections.

Burden and epidemiology of Campylobacter species in acute enteritis cases in Burkina Faso

BackgroundCampylobacter spp. is a significant etiological agent of bacterial gastroenteritis globally. In Burkina Faso (BFA), the actual impact of this pathogen on gastroenteritis is considerably underestimated, primarily due to inadequate surveillance systems.ObjectivesThis study aimed to investigate the proportion of Campylobacter species responsible for acute gastroenteritis among patients of all ages in urban and rural areas of BFA, using molecular biology techniques.Study design & methodsBetween 2018 and 2021, faecal specimens were obtained from 1,295 individuals presenting with acute gastroenteritis. These samples underwent screening for the Campylobacter coli/jejuni/lari complex utilizing real-time polymerase chain reaction (PCR) assays. Subsequently, positive samples were subjected to species-level differentiation through the application of species-specific primers.ResultsCampylobacter spp. was detected in 25.0% (324/1,295) of the samples analysed. The majority of positive samples (95%, 308/324) were obtained from children under 5 years of age. Species identification was performed on a subset of 114 isolates, revealing 51 Campylobacter jejuni, 10 Campylobacter coli, and 53 Campylobacter isolates that remained unspeciated.ConclusionsThis study reveals a significant prevalence of Campylobacter species among patients with acute gastroenteritis, with a particularly high incidence observed in children under 5 years of age. Based on these findings, the implementation of routine Campylobacter surveillance in public health laboratories is strongly recommended to better monitor and address this health concern.

Exploring the Role of the Environment as a Reservoir of Antimicrobial-Resistant Campylobacter: Insights from Wild Birds and Surface Waters.

Antimicrobial resistance (AMR) is a growing global health challenge, compromising bacterial infection treatments and necessitating robust surveillance and mitigation strategies. The overuse of antimicrobials in humans and farm animals has made them hotspots for AMR. However, the spread of AMR genes in wildlife and the environment represents an additional challenge, turning these areas into new AMR hotspots. Among the AMR bacteria considered to be of high concern for public health, Campylobacter has been the leading cause of foodborne infections in the European Union since 2005. This study examines the prevalence of AMR genes and virulence factors in Campylobacter isolates from wild birds and surface waters in Luxembourg. The findings reveal a significant prevalence of resistant Campylobacter strains, with 12% of C. jejuni from wild birds and 37% of C. coli from surface waters carrying resistance genes, mainly against key antibiotics like quinolones and tetracycline. This study underscores the crucial role of the environment in the spread of AMR bacteria and genes, highlighting the urgent need for enhanced surveillance and control measures to curb AMR in wildlife and environmental reservoirs and reduce transmission risks to humans. This research supports One Health approaches to tackling antimicrobial resistance and protecting human, animal, and environmental health.