RESUMEN

Campylobacteriosis outbreaks have previously been linked to dairy foods. While the genetic diversity of Campylobacter is well understood in high-income countries, it is largely unknown in low-income countries, such as Ethiopia. This study therefore aimed to conduct the first genomic characterization of Campylobacter isolates from the Ethiopian dairy supply chain to aid in future epidemiological studies. Fourteen C. jejuni and four C. coli isolates were whole genome sequenced using an Illumina platform. Sequences were analyzed using the bioinformatics tools in the GalaxyTrakr platform to identify MLST types, and single nucleotide polymorphisms, and infer phylogenetic relationships among the studied isolates. Assembled genomes were further screened to detect antimicrobial resistance and virulence gene sequences. Among 14 C. jejuni, ST 2084 and ST 51, which belong to the clonal complexes ST-353 and ST-443, respectively, were identified. Among the 4 sequenced C. coli isolates, two isolates belonged to ST 1628 and two to ST 830 from the clonal complex ST-828. The isolates of C. jejuni ST 2084 and ST 51 carried ß-lactam resistance gene blaOXA-605, a fluoroquinolone resistance-associated mutation T86I in the gryA gene, and a macrolide resistance-associated mutation A103V in 50S L22. Only ST 2084 isolates carried the tetracycline resistance gene tetO. Conversely, all four C. coli ST 830 and ST 1628 isolates carried tetO, but only ST 1628 isolates also carried blaOXA-605. Lastly, C. jejuni ST 2084 isolates carried a total of 89 virulence genes, and ST 51 isolates carried up to 88 virulence genes. Among C. coli, ST 830 isolates carried 71 genes involved in virulence, whereas two ST 1628 isolates carried up to 82 genes involved in virulence. Isolates from all identified STs have previously been isolated from human clinical cases, demonstrating a potential food safety concern. This finding warrants further monitoring of Campylobacter in dairy foods in Ethiopia to better understand and manage the risks associated with Campylobacter contamination and transmission.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Variación Genética , Filogenia , Campylobacter coli/genética , Campylobacter coli/aislamiento & purificación , Campylobacter coli/efectos de los fármacos , Campylobacter coli/patogenicidad , Campylobacter jejuni/genética , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/patogenicidad , Etiopía/epidemiología , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/epidemiología , Productos Lácteos/microbiología , Genoma Bacteriano/genética , Secuenciación Completa del Genoma , Polimorfismo de Nucleótido Simple , Microbiología de Alimentos , Antibacterianos/farmacología , Humanos , Tipificación de Secuencias Multilocus , Virulencia/genética , Farmacorresistencia Bacteriana/genética , Animales

RESUMEN

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 16S23S 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.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Fucosa , Glucosa , Campylobacter coli/genética , Campylobacter coli/aislamiento & purificación , Campylobacter coli/metabolismo , Campylobacter jejuni/genética , Campylobacter jejuni/metabolismo , Campylobacter jejuni/aislamiento & purificación , Glucosa/metabolismo , Humanos , Fucosa/metabolismo , Genoma Bacteriano , Transferencia de Gen Horizontal , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/veterinaria , Familia de Multigenes , Finlandia , Países Bajos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo

RESUMEN

Campylobacter spp. genera is one of the most common causes of microbial enteritis worldwide. This study aimed to find out how common Campylobacter organisms were in raw meat from large livestock in Iran, as well as to determine their antibiotic susceptibility profiles. Several 550 fresh, ready-to-eat meat samples were collected from slaughterhouses, butcher shops, and restaurants in the study region. The samples were collected from cattle (n=138), goats (n=102), camels (n=56), and sheep (n=254). Campylobacter spp. were isolated and identified using normal bacteriological methods and polymerase chain reaction (PCR). Genotyping was performed using PCR to identify virulence genes. The disc diffusion technique was used to determine antibiotic susceptibility. The two Campylobacter spp. were found in 84 (15.27%) of the 550 meat samples tested. Cattle and camel samples accounted for the highest (52.38%) and lowest (3.57%) frequencies of Campylobacter spp., respectively. There were significant differences in the prevalence of Campylobacter spp. in cattle (2=43.04 or OR=7.68, CI=3.40-17.30, P<0.01). Campylobacter jejuni and Campylobacter coli accounted for 82.14% (n=69) of Campylobacter spp. isolated from raw meat. While C. jejuni was found in 39.28% of the samples (n=33), C. coli was observed in 42.85% (n=36). Other Campylobacter spp. formed 17.85 % (n=15) of the samples. The most common genotypes observed in C. jejuni bacteria collected from different types of large animal samples were ciaB (100%) and flaA (100%). On the other hand, virbll (7.69%) was the C. jejuni strain found with the lowest incidence in different large animal samples. The most frequent genotypes found in C. coli bacteria were ciaB (100%) and flaA (100%). C. coli isolates dnaJ (0%), wlaN (0%), virbll (0%), and ceuE (0%) were detected with the lowest frequency in several samples from large livestock. Campylobacter spp. isolated from different sample types and sources were 100% sensitive to aphA-3-1 and GM10. The isolates were reported to be resistant to E15 (76.93%), cmeB (69.24%), aadE1 (69.24%), CIP5 (69.24%), and AM10 (69.24%). According to this study, Campylobacter was found in food from factory farming. Consequently, the disease can be transmitted by eating raw or undercooked meat. Therefore, proper handling and preparation of meat meals, as well as hygiene measures from the slaughterhouse to the retailer, are critical in preventing Campylobacter infections.

Asunto(s)

Antibacterianos , Camelus , Campylobacter coli , Campylobacter jejuni , Farmacorresistencia Bacteriana , Cabras , Carne , Animales , Campylobacter jejuni/efectos de los fármacos , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/genética , Irán/epidemiología , Campylobacter coli/efectos de los fármacos , Campylobacter coli/aislamiento & purificación , Bovinos , Ovinos , Camelus/microbiología , Carne/microbiología , Antibacterianos/farmacología , Microbiología de Alimentos , Ganado/microbiología

RESUMEN

Campylobacter is a zoonotic foodborne pathogen that is often linked with gastroenteritis and other extraintestinal infections in humans. This study is aimed at determining the genetic determinants of virulence-encoding genes responsible for flagellin motility protein A (flaA), Campylobacter adhesion to fibronectin F (cadF), Campylobacter invasion antigen B (ciaB) and cytolethal distending toxin (cdt) A (cdtA) in Campylobacter species. A total of 29 Campylobacter coli isolates (16 from cattle, 9 from chicken, and 4 from water samples) and 74 Campylobacter jejuni isolates (38 from cattle, 30 from chicken, and 6 from water samples) described in an earlier study in Kajiado County, Kenya, were examined for the occurrence of virulence-associated genes using polymerase chain reaction (PCR) and amplicon sequencing. The correlations among virulence genes were analyzed using Pearson's correlation coefficient (R) method. Among the 103 Campylobacter strains screened, 89 were found to harbour a single or multiple virulence gene(s), giving an overall prevalence of 86.4%. C. jejuni strains had the highest prevalence of multivirulence at 64.9% (48/74), compared to C. coli (58.6%, 17/29). The ciaB and flaA genes were the most common virulence genes detected in C. jejuni (81.1% [60/74] and 62.2% [46/74], respectively) and in C. coli (each at 62.1%; 18/29). Campylobacter isolates from chicken harboured the most virulence-encoding genes. C. jejuni strains from chicken and cattle harboured the highest proportions of the cdtA and ciaB genes, respectively. All the C. coli strains from water samples harboured the cadF and flaA genes. The results obtained further revealed a significant positive correlation between cadF and flaA (R = 0.733). C. jejuni and C. coli strains from cattle, chicken, and water harbour virulence markers responsible for motility/colonization, invasion, adherence, and toxin production, evoking their important role in campylobacteriosis development among humans and livestock. The identification of cattle, chicken, and water samples as reservoirs of virulent Campylobacter spp. highlights the possible risk to human health. These data on some virulence genes of Campylobacter will assist food safety and public health officials in formulating policy statements.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Pollos , Heces , Animales , Campylobacter jejuni/genética , Campylobacter jejuni/patogenicidad , Campylobacter jejuni/aislamiento & purificación , Pollos/microbiología , Bovinos , Campylobacter coli/genética , Campylobacter coli/patogenicidad , Campylobacter coli/aislamiento & purificación , Virulencia/genética , Heces/microbiología , Kenia/epidemiología , Factores de Virulencia/genética , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/veterinaria , Microbiología del Agua , Flagelina/genética , Humanos , Proteínas Bacterianas/genética

RESUMEN

Campylobacter spp. are leading bacterial gastroenteritis pathogens. Infections are largely underreported, and the burden of outbreaks may be underestimated. Current strategies of testing as few as one isolate per sample can affect attribution of cases to epidemiologically important sources with high Campylobacter diversity, such as chicken meat. Multiple culture method combinations were utilized to recover and sequence Campylobacter from 45 retail chicken samples purchased across Norwich, UK, selecting up to 48 isolates per sample. Simulations based on resampling were used to assess the impact of Campylobacter sequence type (ST) diversity on outbreak detection. Campylobacter was recovered from 39 samples (87%), although only one sample was positive through all broth, temperature, and plate combinations. Three species were identified (Campylobacter jejuni, Campylobacter coli, and Campylobacter lari), and 33% of samples contained two species. Positive samples contained 1-8 STs. Simulation revealed that up to 87 isolates per sample would be required to detect 95% of the observed ST diversity, and 26 isolates would be required for the average probability of detecting a random theoretical outbreak ST to reach 95%. An optimized culture approach and selecting multiple isolates per sample are essential for more complete Campylobacter recovery to support outbreak investigation and source attribution.

Asunto(s)

Campylobacter , Pollos , Pollos/microbiología , Animales , Campylobacter/aislamiento & purificación , Campylobacter/genética , Campylobacter/clasificación , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/veterinaria , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/genética , Campylobacter coli/aislamiento & purificación , Campylobacter coli/genética , Microbiología de Alimentos , Brotes de Enfermedades , Reino Unido/epidemiología , Carne/microbiología , Variación Genética , Campylobacter lari/genética , Campylobacter lari/aislamiento & purificación

RESUMEN

Campylobacter is gram-negative bacteria considered the predominant genera isolated from poultry samples and associated with gastroenteritis. Due to the problems in conventional cultural methods of time-consuming and technically demanding requirements, a rapid and feasible method for their identification and discrimination of the closely related spp. Including Campylobacter coli, Campylobacter fetus, and Campylobacter jejuni is needed. This study analyzes the chicken and sheep meats samples (n = 125) using culture and pre-enrichment-based Quadraplex real-time PCR by targeting OrfA, CstA, HipO, and 16 S rRNA genes of C. coli, C. fetus, C. jejuni and Campylobacter spp. Respectively. The analysis of 125 chicken and sheep meat samples by culture and real-time PCR showed high concordance between the results of the two methods. The present study show high prevalence of Campylobacter species (35% and 32% from chicken and meat respectively) of which C. jejuni were the most abundant. Reaction efficiencies were between 90 and 110%, and detect as low as 8.9 fg in C. jejuni. The need for quick detection and discrimination methods in sheep and chicken meat can be met using the described Quadraplex real-time PCR methodology.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Pollos , Carne , Reacción en Cadena en Tiempo Real de la Polimerasa , Animales , Pollos/microbiología , Ovinos/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Campylobacter coli/genética , Campylobacter coli/aislamiento & purificación , Campylobacter coli/clasificación , Campylobacter jejuni/genética , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/clasificación , Carne/microbiología , Campylobacter fetus/genética , Campylobacter fetus/aislamiento & purificación , Campylobacter fetus/clasificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , Campylobacter/clasificación , Microbiología de Alimentos , ADN Bacteriano/genética

RESUMEN

BACKGROUND: Children with under-five year age disproportionally affected with foodborne illness. Campylobacteriosis is the most common foodborne disease next to Norovirus infection. Macrolides are commonly prescribed as the first line of treatment for Campylobacter gastroenteritis, with fluoroquinolone and tetracycline as secondary options. However, resistance to these alternatives has been reported in various regions worldwide. OBJECTIVE: To determine the prevalence, associated risk-factors and antimicrobial resistance of Campylobacter jejuni and C. coli among under-five children with diarrhea. METHODS: Institution-based cross-sectional study was conducted from November, 2022 to April 2023. The study sites were selected using a random sampling technique, while the study subjects were included using a convenient sampling technique. The data were collected using a structured questionnaire. Stool samples were inoculated onto modified charcoal cefoperazone deoxycholate agar and incubated for 48 hours. The suspected colonies were analyzed using matrix-assisted laser desorption ionization-time of flight mass spectrometry to confirm the species. Antimicrobial susceptibility testing was performed using a disc diffusion technique. All potential covariates (independent variables) were analyzed one by one using bivariate logistic regression model to identify candidate variables with P value < 0.25. Multivariable logistic analysis was used to identify potential associated factors using the candidate variables. A p value ≤ 0.05 at a 95% confidence interval was statistically significant. RESULT: Among the 428 samples, 7.0% (CI: 4.5-9.3) were confirmed Campylobacter species. The prevalence of C. jejuni and C. coli among under-five children was 5.1% (CI: 3.0-7.0) and 1.9% (CI: 0.7-3.3), respectively. C. jejuni (73.3%) was dominant over C. coli (26.7%). The resident, contact with domestic animals, and parents/guardians education level were significantly associated with campylobacteriosis among under-five children. One-third of the Campylobacter isolates (33.3%, 10/30) were resistant to ciprofloxacin and tetracycline whereas 10.0% (3/30) were resistant to erythromycin. Furthermore, 3.3% (1/30) of the Campylobacter were found to be multidrug-resistant. CONCLUSION: The prevalence of Campylobacter species was 7.0%. The resistance rate of Campylobacter species of ciprofloxacin and tetracycline-resistance strains was 33.3%. Peri-urban residence, contact with domestic animals, and low parental educational statuses were significantly associated factors with increased risk of Campylobacter infection. Continuous surveillance on antimicrobial resistance and health education of personal and environmental hygiene should be implemented in the community.

Asunto(s)

Antibacterianos , Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Diarrea , Humanos , Campylobacter jejuni/efectos de los fármacos , Campylobacter jejuni/aislamiento & purificación , Campylobacter coli/efectos de los fármacos , Campylobacter coli/aislamiento & purificación , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/tratamiento farmacológico , Infecciones por Campylobacter/microbiología , Preescolar , Lactante , Femenino , Masculino , Diarrea/epidemiología , Diarrea/microbiología , Diarrea/tratamiento farmacológico , Etiopía/epidemiología , Estudios Transversales , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana , Prevalencia , Pruebas de Sensibilidad Microbiana , Recién Nacido , Factores de Riesgo

RESUMEN

OBJECTIVES: Campylobacter is a significant zoonotic pathogen primarily transmitted through poultry. Our study aimed to assess antimicrobial resistance and genetic relationships among Campylobacter isolates from retail chicken meat and humans in Taiwan. METHODS: Campylobacter isolates were analysed using whole-genome sequencing to investigate their antimicrobial resistance, genetic determinants of resistance, and genotypes. RESULTS: Campylobacter coli and Campylobacter jejuni accounted for 44.9% and 55.1% of chicken meat isolates, and 11.4% and 88.6% of human isolates, respectively. C. coli displayed significantly higher resistance levels. Furthermore, isolates from chicken meat exhibited higher levels of resistance to most tested antimicrobials compared to isolates from humans. Multidrug resistance was observed in 96.3% of C. coli and 43.3% of C. jejuni isolates from chicken meat and 80.6% of C. coli and 15.8% of C. jejuni isolates from humans. Macrolide resistance was observed in 85.5% of C. coli isolates, primarily attributed to the erm(B) rather than the A2075G mutation in 23S rRNA. Among the 511 genomes, we identified 133 conventional MLST sequence types, indicating significant diversity among Campylobacter strains. Notably, hierarchical Core-genome multilocus sequence typing clustering, including HC0, HC5, and HC10, revealed a significant proportion of closely related isolates from chicken meat and humans. CONCLUSIONS: Our research highlights significant associations in antimicrobial resistance and genetic relatedness between Campylobacter isolates from chicken meat and humans in Taiwan. The genetic analysis data suggest that campylobacteriosis outbreaks may occur more frequently in Taiwan than previously assumed. Our study emphasizes the need for strategies to control multidrug-resistant strains and enhance outbreak prevention.

Asunto(s)

Antibacterianos , Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Pollos , Farmacorresistencia Bacteriana Múltiple , Carne , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Secuenciación Completa del Genoma , Campylobacter jejuni/genética , Campylobacter jejuni/efectos de los fármacos , Campylobacter jejuni/aislamiento & purificación , Campylobacter coli/efectos de los fármacos , Campylobacter coli/genética , Campylobacter coli/aislamiento & purificación , Pollos/microbiología , Animales , Humanos , Taiwán , Antibacterianos/farmacología , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/epidemiología , Carne/microbiología , Farmacorresistencia Bacteriana Múltiple/genética , Genotipo , Farmacorresistencia Bacteriana/genética , Microbiología de Alimentos

RESUMEN

Human gastrointestinal infections caused by Campylobacter species is the second most important foodborne illness after salmonellosis worldwide. Poultry represent one of the main sources of Campylobacter organisms. In the present study, the short variable region of flagellin gene (SVR-flaA) typing was carried out to determine the variation among the circulating strains of Campylobacter jejuni and Campylobacter coli. The C. jejuni and C. coli isolated from poultry and poultry meat were screened for the presence of virulence determinants like cadF, flaA, cdtB, and wlaN gene. The screening for wlaN gene is crucial in view of the fact that most patients with Guillian Barre's (GB) syndrome with a preceding history of diarrheal illness have been found to harbor wlaN gene-positive C jejuni strains. Out of the 200 samples comprising poultry meat and cloacal swabs, 21.5% of samples were found to harbor Campylobacter spp. of which 2.5% were Campylobacter jejuni, and 19% were confirmed as Campylobacter coli. The cadF, flaA, cdtB virulence genes were detected in all the Campylobacter spp. isolated in the present study. The presence of the wlaN gene in the Campylobacter jejuni isolated in the present study may pose a public health threat with long-term human health implications. The SVR-flaA typing of twelve Campylobacter isolates obtained in the present study revealed that Campylobacter coli flaA sequence OL471375 is a new strain with a novel allele type 1,675 and peptide sequence 5 which stands deposited in pubMLST database for Campylobacter. The other flaA-SVR gene sequences identified in this study were OL471369, OL471370, OL471371, OL471372, OL471373, and OL471374. Among twelve Campylobacter spp., three distinct DdeI-RFLP patterns were observed, each varying in size from 100 to 1,000 base pairs. Antimicrobial profiling of the Campylobacter spp. isolated in the present study revealed that 50% of the strains were multidrug resistant. All the Campylobacter spp. were resistant to ciprofloxacin (CIP), ampicillin (AMP), penicillin (PEN), and nalidixic acid (NAL) whereas 57.1% of strains were resistant to tetracycline (TET) and erythromycin (ERY) 28% to amoxicillin (AMX) and enrofloxacin (ENO), 85% to amikacin (AMK). The high degree of resistance to fluoroquinolones observed in the present study is crucial in view of fluoroquinolones being drugs of choice for the treatment of human Campylobacter infections.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Farmacorresistencia Bacteriana Múltiple , Flagelina , Aves de Corral , Animales , Flagelina/genética , Humanos , Campylobacter jejuni/efectos de los fármacos , Campylobacter jejuni/genética , India , Campylobacter coli/efectos de los fármacos , Campylobacter coli/genética , Virulencia , Contaminación de Alimentos/análisis , Microbiología de Alimentos , Factores de Virulencia/genética , Campylobacter/efectos de los fármacos , Campylobacter/genética , Carne/microbiología , Variación Genética , Antibacterianos/farmacología , Pruebas de Sensibilidad Microbiana

RESUMEN

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 ΔoptrA 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 ΔoptrA 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 blaOXA-61. As a result, the 29 ΔoptrA-positive strains displayed susceptibility to both florfenicol and linezolid. The ΔoptrA gene is linked to the erm(A) gene, resulting in the formation of translocatable unit (TU) that are encompassed by two copies of IS1216 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.

Asunto(s)

Antibacterianos , Infecciones por Campylobacter , Campylobacter coli , Pollos , Farmacorresistencia Bacteriana Múltiple , Islas Genómicas , Campylobacter coli/genética , Campylobacter coli/efectos de los fármacos , Islas Genómicas/genética , Pollos/microbiología , Animales , Farmacorresistencia Bacteriana Múltiple/genética , Antibacterianos/farmacología , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/veterinaria , Estudios Retrospectivos , Proteínas Bacterianas/genética , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Enfermedades de las Aves de Corral/microbiología , Polimorfismo de Nucleótido Simple

RESUMEN

Recombination of short DNA fragments via horizontal gene transfer (HGT) can introduce beneficial alleles, create genomic disharmony through negative epistasis, and create adaptive gene combinations through positive epistasis. For non-core (accessory) genes, the negative epistatic cost is likely to be minimal because the incoming genes have not co-evolved with the recipient genome and are frequently observed as tightly linked cassettes with major effects. By contrast, interspecific recombination in the core genome is expected to be rare because disruptive allelic replacement is likely to introduce negative epistasis. Why then is homologous recombination common in the core of bacterial genomes? To understand this enigma, we take advantage of an exceptional model system, the common enteric pathogens Campylobacter jejuni and C. coli that are known for very high magnitude interspecies gene flow in the core genome. As expected, HGT does indeed disrupt co-adapted allele pairings, indirect evidence of negative epistasis. However, multiple HGT events enable recovery of the genome's co-adaption between introgressing alleles, even in core metabolism genes (e.g., formate dehydrogenase). These findings demonstrate that, even for complex traits, genetic coalitions can be decoupled, transferred, and independently reinstated in a new genetic background-facilitating transition between fitness peaks. In this example, the two-step recombinational process is associated with C. coli that are adapted to the agricultural niche.IMPORTANCEGenetic exchange among bacteria shapes the microbial world. From the acquisition of antimicrobial resistance genes to fundamental questions about the nature of bacterial species, this powerful evolutionary force has preoccupied scientists for decades. However, the mixing of genes between species rests on a paradox: 0n one hand, promoting adaptation by conferring novel functionality; on the other, potentially introducing disharmonious gene combinations (negative epistasis) that will be selected against. Taking an interdisciplinary approach to analyze natural populations of the enteric bacteria Campylobacter, an ideal example of long-range admixture, we demonstrate that genes can independently transfer across species boundaries and rejoin in functional networks in a recipient genome. The positive impact of two-gene interactions appears to be adaptive by expanding metabolic capacity and facilitating niche shifts through interspecific hybridization. This challenges conventional ideas and highlights the possibility of multiple-step evolution of multi-gene traits by interspecific introgression.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Epistasis Genética , Transferencia de Gen Horizontal , Genoma Bacteriano , Recombinación Genética , Campylobacter jejuni/genética , Campylobacter coli/genética , Evolución Molecular , Adaptación Fisiológica/genética , Adaptación Biológica/genética

RESUMEN

Campylobacter causes bacterial enteritis, dysentery, and growth faltering in children in low- and middle-income countries (LMICs). Campylobacter spp. are fastidious organisms, and their detection often relies on culture independent diagnostic technologies, especially in LMICs. Campylobacter jejuni and Campylobacter coli are most often the infectious agents and in high income settings together account for 95% of Campylobacter infections. Several other Campylobacter species have been detected in LMIC children at an increased prevalence relative to high income settings. After doing extensive whole genome sequencing of isolates of C. jejuni and C. coli in Peru, we observed heterogeneity in the binding sites for the main species-specific PCR assay (cadF) and designed an alternative rpsKD-based qPCR assay to detect both C. jejuni and C. coli. The rpsKD-based qPCR assay identified 23% more C.jejuni/ C.coli samples than the cadF assay among 47 Campylobacter genus positive cadF negative samples verified to have C. jejuni and or C. coli with shotgun metagenomics. This assay can be expected to be useful in diagnostic studies of enteric infectious diseases and be useful in revising the attribution estimates of Campylobacter in LMICs.

Asunto(s)

Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Campylobacter , Niño , Humanos , Campylobacter coli/genética , Reacción en Cadena de la Polimerasa , Infecciones por Campylobacter/diagnóstico , Infecciones por Campylobacter/microbiología , Heces/microbiología

RESUMEN

BACKGROUND: Campylobacter jejuni and Campylobacter coli are the major causative agents of bacterial gastroenteritis worldwide and are known obligate microaerophiles. Despite being sensitive to oxygen and its reduction products, both species are readily isolated from animal food products kept under atmospheric conditions where they face high oxygen tension levels. RESULTS: In this study, Transposon Directed Insertion-site Sequencing (TraDIS) was used to investigate the ability of one C. jejuni strain and two C. coli strains to overcome oxidative stress, using H2O2 to mimic oxidative stress. Genes were identified that were required for oxidative stress resistance for each individual strain but also allowed a comparison across the three strains. Mutations in the perR and ahpC genes were found to increase Campylobacter tolerance to H2O2. The roles of these proteins in oxidative stress were previously known in C. jejuni, but this data indicates that they most likely play a similar role in C. coli. Mutation of czcD decreased Campylobacter tolerance to H2O2. The role of CzcD, which functions as a zinc exporter, has not previously been linked to oxidative stress. The TraDIS data was confirmed using defined deletions of perR and czcD in C. coli 15-537360. CONCLUSIONS: This is the first study to investigate gene fitness in both C. jejuni and C. coli under oxidative stress conditions and highlights both similar roles for certain genes for both species and highlights other genes that have a role under oxidative stress.

Asunto(s)

Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Animales , Campylobacter jejuni/genética , Campylobacter jejuni/metabolismo , Campylobacter coli/genética , Campylobacter coli/metabolismo , Peróxido de Hidrógeno/farmacología , Peróxido de Hidrógeno/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Estrés Oxidativo/genética , Oxígeno/metabolismo , Infecciones por Campylobacter/microbiología

RESUMEN

Chickens are the primary reservoirs of Campylobacter spp., mainly C. jejuni and C. coli, that cause human bacterial gastrointestinal infections. However, genomic characteristics and antimicrobial resistance of Campylobacter spp. in low- to middle-income countries need more comprehensive exploration. This study aimed to characterize 21 C. jejuni and 5 C. coli isolates from commercial broilers and native chickens using whole genome sequencing and compare them to 28 reference Campylobacter sequences. Among the 26 isolates, 13 sequence types (ST) were identified in C. jejuni and 5 ST in C. coli. The prominent ST was ST 2274 (5 isolates, 19.2%), followed by ST 51, 460, 2409, and 6455 (2 isolates in each ST, 7.7%), while all remaining ST (464, 536, 595, 2083, 6736, 6964, 8096, 10437, 828, 872, 900, 8237, and 13540) had 1 isolate per ST (3.8%). Six types of antimicrobial resistance genes (ant(6)-Ia, aph(3')-III, blaOXA, cat, erm(B), and tet(O)) and one point mutations in the gyrA gene (Threonine-86-Isoleucine) and another in the rpsL gene (Lysine-43-Arginine) were detected. The blaOXA resistance gene was present in all isolates, the gyrA mutations was in 95.2% of C. jejuni and 80.0% of C. coli, and the tet(O) resistance gene in 76.2% of C. jejuni and 80.0% of C. coli. Additionally, 203 virulence-associated genes linked to 16 virulence factors were identified. In terms of phenotypic resistance, the C. jejuni isolates were all resistant to ciprofloxacin, enrofloxacin, and nalidixic acid, with lower levels of resistance to tetracycline (76.2%), tylosin (52.3%), erythromycin (23.8%), azithromycin (22.2%), and gentamicin (11.1%). Most C. coli isolates were resistant to all tested antimicrobials, while 1 C. coli was pan-susceptible except for tylosin. Single-nucleotide polymorphisms concordance varied widely, with differences of up to 13,375 single-nucleotide polymorphisms compared to the reference Campylobacter isolates, highlighting genetic divergence among comparative genomes. This study contributes to a deeper understanding of the molecular epidemiology of Campylobacter spp. in Thai chicken production systems.

Asunto(s)

Antiinfecciosos , Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Campylobacter , Animales , Humanos , Pollos/genética , Tailandia/epidemiología , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/veterinaria , Infecciones por Campylobacter/microbiología , Tilosina , Farmacorresistencia Bacteriana/genética , Campylobacter/genética , Antibacterianos/farmacología , Secuenciación Completa del Genoma/veterinaria , Pruebas de Sensibilidad Microbiana/veterinaria

RESUMEN

Foodborne infections with antimicrobial-resistant Campylobacter spp. remain an important public health concern. Publicly available data collected by the National Antimicrobial Resistance Monitoring System for Enteric Bacteria related to antimicrobial resistance (AMR) in Campylobacter spp. isolated from broiler chickens and turkeys at the slaughterhouse level across the United States between 2013 and 2021 were analysed. A total of 1,899 chicken-origin (1,031 Campylobacter coli (C. coli) and 868 Campylobacter jejuni (C. jejuni)) and 798 turkey-origin (673 C. coli and 123 C. jejuni) isolates were assessed. Chicken isolates exhibited high resistance to tetracycline (43.65%), moderate resistance to ciprofloxacin (19.5%), and low resistance to clindamycin (4.32%) and azithromycin (3.84%). Turkey isolates exhibited very high resistance to tetracycline (69%) and high resistance to ciprofloxacin (39%). The probability of resistance to all tested antimicrobials, except for tetracycline, significantly decreased during the latter part of the study period. Turkey-origin Campylobacter isolates had higher odds of resistance to all antimicrobials than isolates from chickens. Compared to C. jejuni isolates, C. coli isolates had higher odds of resistance to all antimicrobials, except for ciprofloxacin. The study findings emphasize the need for poultry-type-specific strategies to address differences in AMR among Campylobacter isolates.

Asunto(s)

Antiinfecciosos , Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Campylobacter , Animales , Estados Unidos/epidemiología , Antibacterianos/farmacología , Pollos/microbiología , Pavos/microbiología , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana , Ciprofloxacina/farmacología , Tetraciclina/farmacología , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/veterinaria , Infecciones por Campylobacter/microbiología

RESUMEN

Campylobacter spp. are a common cause of bacterial gastroenteritis in Australia, primarily acquired from contaminated meat. We investigated the relationship between genomic virulence characteristics and the severity of campylobacteriosis, hospitalisation, and other host factors.We recruited 571 campylobacteriosis cases from three Australian states and territories (2018-2019). We collected demographic, health status, risk factors, and self-reported disease data. We whole genome sequenced 422 C. jejuni and 84 C. coli case isolates along with 616 retail meat isolates. We classified case illness severity using a modified Vesikari scoring system, performed phylogenomic analysis, and explored risk factors for hospitalisation and illness severity.On average, cases experienced a 7.5 day diarrhoeal illness with additional symptoms including stomach cramps (87.1 %), fever (75.6 %), and nausea (72.0 %). Cases aged ≥75 years had milder symptoms, lower Vesikari scores, and higher odds of hospitalisation compared to younger cases. Chronic gastrointestinal illnesses also increased odds of hospitalisation. We observed significant diversity among isolates, with 65 C. jejuni and 21 C. coli sequence types. Antimicrobial resistance genes were detected in 20.4 % of isolates, but multidrug resistance was rare (0.04 %). Key virulence genes such as cdtABC (C. jejuni) and cadF were prevalent (>90 % presence) but did not correlate with disease severity or hospitalisation. However, certain genes (e.g. fliK, Cj1136, and Cj1138) appeared to distinguish human C. jejuni cases from food source isolates.Campylobacteriosis generally presents similarly across cases, though some are more severe. Genotypic virulence factors identified in the literature to-date do not predict disease severity but may differentiate human C. jejuni cases from food source isolates. Host factors like age and comorbidities have a greater influence on health outcomes than virulence factors.

Asunto(s)

Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Gastroenteritis , Humanos , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/microbiología , Campylobacter coli/genética , Australia/epidemiología , Factores de Virulencia/genética , Genómica

RESUMEN

RESEARCH HIGHLIGHTS: High Campylobacter prevalence in chickens; C. jejuni more prevalent than C. coli.Susceptibility to macrolides but resistance to quinolones/tetracyclines in isolates.Homogeneous resistance patterns within farms; higher in broilers than in native birds.Partial association between phenotypic and genotypic resistance among isolates.

Asunto(s)

Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Campylobacter , Animales , Pollos , Campylobacter jejuni/genética , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/veterinaria , Tailandia/epidemiología , Antibacterianos/farmacología , Campylobacter coli/genética , Farmacorresistencia Bacteriana/genética , Pruebas de Sensibilidad Microbiana/veterinaria

RESUMEN

In this study, we aimed to determine the antibiotic resistance status of Campylobacter spp. isolated from human infections in our region, including the role of mechanisms involved in erythromycin resistance. Standard methods were used for the isolation, identification and antibiotic susceptibility testing of Campylobacter spp. isolates. Erythromycin-resistant mutants were selected from erythromycin-susceptible clinical isolates, and the erythromycin resistance mechanisms were investigated phenotypically by determining the erythromycin MICs of isolates in the presence and absence of the resistance nodulation cell division (RND) type efflux pump inhibitor, phenylalanine-arginine ß-naphthylamide dihydrochloride (PAßN), and genotypically by determining ribosomal and cmeABC alterations using PCR and DNA sequence analysis. Campylobacter spp., including 184 C. jejuni and 20 C. coli in a two-year period, were the most frequently isolated gastrointestinal bacterial pathogens in our region. However, in both C. jejuni and C. coli, resistance to tetracycline and ciprofloxacin were found to be high, erythromycin resistance was especially high (20%) in C. coli. With a ribosomal alteration, A2075G, which was found to be associated with high-level erythromycin resistance in clinical isolates, PAßN significantly reduced the erythromycin MICs in both clinical isolates and mutants. An important finding of this study, while considering cmeABC operon, is the explanation of why erythromycin resistance is more common among C. coli than C. jejuni, bearing in mind the specific deletions and alterations in the intergenic region of the operon in all erythromycin-resistant C. coli isolates. Ultimately, these findings revealed the significant role of RND-type efflux activity in increased erythromycin MICs of the isolates.

Asunto(s)

Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Humanos , Eritromicina/farmacología , Campylobacter jejuni/genética , Campylobacter coli/genética , Farmacorresistencia Bacteriana/genética , Antibacterianos/farmacología , Pruebas de Sensibilidad Microbiana , División Celular , Infecciones por Campylobacter/microbiología

RESUMEN

BACKGROUND: Early childhood enteric infection with Shigella/EIEC, enteroaggregative E. coli (EAEC), Campylobacter, and Giardia has been associated with reduced child growth, yet a recent randomized trial of antimicrobial therapy to reduce these infections did not improve growth outcomes. To interrogate this discrepancy, we measured the enteric infections from this study. METHODS: We leveraged the Early Life Interventions for Childhood Growth and Development in Tanzania (ELICIT) trial, a randomized double-blind placebo-controlled trial of antimicrobial therapy with azithromycin and nitazoxanide provided quarterly to infants from 6 to 15 months of age. We tested 5,479 stool samples at time points across the study for 34 enteropathogens using quantitative PCR. RESULTS: There was substantial carriage of enteropathogens in stool. Azithromycin administration led to reductions in Campylobacter jejuni/coli, enteroaggregative E. coli, and Shigella/EIEC (absolute risk difference ranged from -0.06 to 0.24) 2 weeks after treatment however there was no effect after 3 months. There was no difference in Giardia after nitazoxanide administration (ARR 0.03 at the 12 month administration). When examining the effect of azithromycin versus placebo on the subset of children infected with specific pathogens at the time of treatment, a small increase in weight-for-age Z score was seen only in those infected with Campylobacter jejuni/coli (0.10 Z score, 95% CI -0.01-0.20; length-for-age Z score 0.07, 95% CI -0.06-0.20). CONCLUSION: The antimicrobial intervention of quarterly azithromycin plus or minus nitazoxanide led to only transient decreases in enteric infections with Shigella/EIEC, enteroaggregative E. coli (EAEC), Campylobacter, and Giardia. There was a trend towards improved growth in children infected with Campylobacter that received quarterly azithromycin.

Asunto(s)

Antiinfecciosos , Campylobacter coli , Campylobacter , Lactante , Niño , Humanos , Preescolar , Azitromicina/uso terapéutico , Escherichia coli , Tanzanía , Crecimiento y Desarrollo , Diarrea/tratamiento farmacológico , Heces

RESUMEN

AIMS: To evaluate the biofilm-forming capacity of thermotolerant Campylobacter (TC) strains from poultry production and to analyse the inhibitory capacity of Lactiplantibacillus plantarum LP5 against TC on different materials. METHODS AND RESULTS: Biofilm-forming capacity by Campylobacter jejuni and Campylobacter coli was analysed by cell adhesion in polystyrene plates. TC were classified as non-biofilm-forming (NBF, 1.3%), weak biofilm-forming (WBF, 68.4%), moderate biofilm-forming (MBF, 27.6%), and strong biofilm-forming (SBF, 2.7%). The inhibitory capacity of L. plantarum LP5 against TC was tested on stainless-steel, nylon, aluminium, and glass disks (treated group) and compared with biofilm-forming TC (control group). Lactiplantibacillus plantarum LP5 was inoculated, and then TC. Biofilm was removed in both experimental groups and TC and LP5 bacterial counts were performed. The L. plantarum LP5 presence reduced the formation of TC biofilm (P < 0.001). The material type and strain category influenced biofilm formation, with stainless-steel and the SBF strain being the material and TC having the highest adhesion (P < 0.001). Lactiplantibacillus plantarum LP5 formed a similar biofilm on all materials (P = 0.823). CONCLUSIONS: This trial showed very promising results; L. plantarum LP5 could be incorporated as a bio-protector of TC on different surfaces.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Campylobacter , Lactobacillus plantarum , Biopelículas , Acero