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1.
Int J Infect Dis ; 144: 107055, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38723889

ABSTRACT

OBJECTIVES: To investigate cases of five Campylobacter jejuni outbreaks and describe laboratory characteristics of these infections. METHODS: Whole-genome sequencing and conventional methods were combined to thoroughly investigate the outbreaks, and data of contemporaneous sporadic cases was included for comparison. RESULTS: Seven sequence types (STs) of C. jejuni caused 83 cases, including ST9079 which recurred across 2 years. Trace-back investigation could not identify any food items of infection but detected identical campylobacters from food contacts. Phylogenetic analysis unveiled genetic closeness between outbreak strains and some concurrent sporadic strains, indicating local campylobacteriosis may not be wholly sporadic but rather a series of linked cases. Virulence genes disclosed species/case-specific signatures to differentiate outbreaks from truly non-outbreak strains. Resistance to fluoroquinolones and/or macrolides was prevalent (90.8%, 108/119), with a noteworthy portion exhibiting multidrug resistance (31.1%, 37/119). Five types of plasmids were harbored among outbreak isolates, of which one plasmid harboring anti-stress and resistant genes was rarely found in C. jejuni. CONCLUSIONS: This is the first reported sequential outbreak of C. jejuni in China. Our observations help to define the genomic landscape and antimicrobial resistance patterns of Campylobacter, emphasizing the need for a broader 'One Health' perspective to combat the threats posed by campylobacteriosis.


Subject(s)
Anti-Bacterial Agents , Campylobacter Infections , Campylobacter jejuni , Disease Outbreaks , Phylogeny , Whole Genome Sequencing , Campylobacter jejuni/genetics , Campylobacter jejuni/drug effects , Campylobacter jejuni/isolation & purification , Campylobacter jejuni/classification , Humans , China/epidemiology , Campylobacter Infections/epidemiology , Campylobacter Infections/microbiology , Adult , Child , Male , Female , Anti-Bacterial Agents/pharmacology , Adolescent , Middle Aged , Drug Resistance, Multiple, Bacterial/genetics , Young Adult , Child, Preschool , Microbial Sensitivity Tests , Plasmids/genetics , Genome, Bacterial
2.
Open Vet J ; 14(3): 759-768, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38682147

ABSTRACT

Background: Poultry is one of the most prominent sources of Campylobacter jejuni, which is also a major means of transmission to people. Campylobacter jejuni contamination in chicken meat comes from chicken feces because it naturally exists in the intestines of chickens. Aim: The purpose of this study is to identify the antibiotic resistance patterns and genes of C. jejuni, which was found in chickens in Pasuruan, Indonesia. Methods: The samples used in this study were 200 contents of the small intestine of broiler chickens from 40 farms in Pasuruan Regency. The enriched sample was streaked on the selective media of modified charcoal cefoperazone deoxycholate agar containing the CCDA selective supplement. Antimicrobial susceptibility test utilizing the Kirby-Bauer diffusion test method in accordance with Clinical and Laboratory Standards Institute standards. The polymerase chain reaction (PCR) method was used to detect the (hipO), which encodes the C. jejuni strain, fluoroquinolone resistance (gyrA), beta-lactam resistance (blaOXA-61), and tetracycline resistance (tetO) genes. Results: The findings revealed a 14% (28/200) prevalence of C. jejuni in the small intestine of broiler chickens. These isolates showed high resistance to enrofloxacin (92.9%). All isolates (100%) were susceptible to amoxicillin-clavulanate. The PCR results showed all C. jejuni isolates (100%) detected the gyrA gene, 96.4% detected the blaOXA-61 gene, and 50% detected the tetO gene. Conclusion: The findings of antimicrobial resistance at a high level from the small intestine of broiler chickens illustrate the potential threat to human health. To lessen the effects now and in the future, coordinated and suitable action is needed, as well as steps to guarantee the poultry industry's economic survival and public health insurance.


Subject(s)
Anti-Bacterial Agents , Campylobacter Infections , Campylobacter jejuni , Chickens , Drug Resistance, Bacterial , Poultry Diseases , Animals , Campylobacter jejuni/drug effects , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Chickens/microbiology , Indonesia/epidemiology , Campylobacter Infections/veterinary , Campylobacter Infections/microbiology , Campylobacter Infections/epidemiology , Poultry Diseases/microbiology , Poultry Diseases/epidemiology , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/genetics , Microbial Sensitivity Tests/veterinary
3.
Eur J Clin Microbiol Infect Dis ; 43(5): 895-904, 2024 May.
Article in English | MEDLINE | ID: mdl-38472522

ABSTRACT

PURPOSE: Campylobacter is a frequent cause of enteric infections with common antimicrobial resistance issues. The most recent reports of campylobacteriosis in Italy include data from 2013 to 2016. We aimed to provide national epidemiological and microbiological data on human Campylobacter infections in Italy during the period 2017-2021. METHODS: Data was collected from 19 Hospitals in 13 Italian Regions. Bacterial identification was performed by mass spectrometry. Antibiograms were determined with Etest or Kirby-Bauer (EUCAST criteria). RESULTS: In total, 5419 isolations of Campylobacter spp. were performed. The most common species were C. jejuni (n = 4535, 83.7%), followed by C. coli (n = 732, 13.5%) and C. fetus (n = 34, 0.6%). The mean age of patients was 34.61 years and 57.1% were males. Outpatients accounted for 54% of the cases detected. Campylobacter were isolated from faeces in 97.3% of cases and in 2.7% from blood. C. fetus was mostly isolated from blood (88.2% of cases). We tested for antimicrobial susceptibility 4627 isolates (85.4%). Resistance to ciprofloxacin and tetracyclines was 75.5% and 54.8%, respectively; resistance to erythromycin was 4.8%; clarithromycin 2% and azithromycin 2%. 50% of C. jejuni and C. coli were resistant to ≥ 2 antibiotics. Over the study period, resistance to ciprofloxacin and tetracyclines significantly decreased (p < 0.005), while resistance to macrolides remained stable. CONCLUSION: Campylobacter resistance to fluoroquinolones and tetracyclines in Italy is decreasing but is still high, while macrolides retain good activity.


Subject(s)
Anti-Bacterial Agents , Campylobacter Infections , Campylobacter , Microbial Sensitivity Tests , Humans , Campylobacter Infections/epidemiology , Campylobacter Infections/microbiology , Italy/epidemiology , Female , Male , Adult , Anti-Bacterial Agents/pharmacology , Middle Aged , Young Adult , Adolescent , Aged , Campylobacter/drug effects , Campylobacter/isolation & purification , Child , Child, Preschool , Infant , Feces/microbiology , Drug Resistance, Bacterial , Aged, 80 and over , Infant, Newborn , Campylobacter jejuni/drug effects , Campylobacter jejuni/isolation & purification
4.
Pediatr. aten. prim ; 25(99): e91-e94, 3 oct. 2023.
Article in Spanish | IBECS | ID: ibc-226243

ABSTRACT

Introducción: Campylobacter es el principal patógeno de gastroenteritis transmitida por alimentos, ocurriendo generalmente por la ingesta de pollo mal cocinado, constituyendo otra importante fuente de infección los cachorros de animales domésticos. Caso clínico: escolar con gastroenteritis aguda con sospecha diagnóstica inicial de giardiasis por ambiente epidémico (gato doméstico). Se recoge coprocultivo en el que se detecta Campylobacter jejuni, prescribiéndose azitromicina, dado lo prolongado de la clínica. A lo largo del control evolutivo en el centro de salud la familia informa de que se ha solicitado nueva muestra de heces en el gato, dado persistencia de los síntomas pese a tratamiento con metronidazol. Finalmente, crece también Campylobacter jejuni en el coprocultivo de la mascota. Tras finalizar ambos el tratamiento antibiótico, permanecen asintomáticos. Como posible alimento sospechoso del origen del cuadro está el corazón de pollo no cocinado con el que alimentaban al gato de forma habitual. Conclusiones: ante un cuadro de gastroenteritis aguda es fundamental una adecuada anamnesis que incluya ambiente epidémico y alimentos sospechosos. En ocasiones las mascotas también constituyen una fuente de transmisión de la infección a nuestros pacientes. En este caso se sospecha la cadena de contaminación: corazón de pollo no cocinado-heces de gato doméstico-niña (AU)


Introduction: Campylobacter is a well-known food-borne pathogen that causes human gastroenteritis. The most common way for children to become infected with campylobacteriosis is through chicken that is not fully cooked, another important source of infection are domestic puppies.Case report: it is presented the case of an eight-year-old girl with acute gastroenteritis, the first diagnostic suspicion was giardiasis due to epidemic environment (domestic cat). A stool culture was collected in which Campylobacter jejuni was detected. Azithromycin was prescribed because of prolonged symptoms. Throughout the control in the health center, family reported that a new fecal sample has been requested from the cat due to the persistence of the symptoms despite treatment with metronidazole. Finally, Campylobacter jejuni also grew in the pet's stool culture. After both finished antibiotic treatment, they remained asymptomatic. The possible suspected infection source was the chicken heart with which the cat was regularly fed. Conclusions: the evaluation of the child with acute gastroenteritis begins with a careful history which includes epidemiological environment and suspicious food intake. Ocassionally, pets are also a source of transmission to our patients. In this case, the suspected contamination chain was: uncooked chicken heart- domestic cat faeces-girl. (AU)


Subject(s)
Humans , Female , Child, Preschool , Campylobacter jejuni/isolation & purification , Campylobacter Infections/diagnosis , Campylobacter Infections/drug therapy , Gastroenteritis/drug therapy , Gastroenteritis/microbiology , Dysentery/drug therapy , Dysentery/microbiology
6.
Biosci Rep ; 41(9)2021 09 30.
Article in English | MEDLINE | ID: mdl-34519329

ABSTRACT

Campylobacter jejuni and Salmonella typhimurium are the leading causes of bacterial food contamination in chicken carcasses. Contamination is particularly associated with the slaughtering process. The present study isolated C. jejuni and S. typhimurim from fifty chicken carcass samples, all of which were acquired from different companies in Riyadh, Saudi Arabia. The identification of C. jejuni was performed phenotypically by using a hippurate test and genetically using a polymerase chain reaction with primers for 16S rRNA and hippurate hydrolase (hipO gene). For the dentification of S. typhimurim, a serological Widal test was carried out using serum anti-S. typhimurium antibodies. Strains were genetically detected using invA gene primers. The positive isolates for C. jejuni showed a specific molecular size of 1448 bp for 16S rRNA and 1148 bp for hipO genes. However, the positive isolates of the invA gene exhibited a specific molecular size at 244 bp using polymerase chain reaction (PCR). Comparing sequencing was performed with respect to the invA gene and the BLAST nucleotide isolates that were identified as Salmonella enterica subsp. enterica serovar typhimurium strain ST45, thereby producing a similarity of 100%. The testing identified C.jejuni for hippuricase, GenBank: Z36940.1. While many isolates of Salmonella spp. that contained the invA gene were not necessarily identified as S. typhimurim, the limiting factor for the Widal test used antiS. typhimurum antibodies. The multidrug resistance (MDR) of C. jejuni isolates in chickens was compared with the standard C. jejuni strain ATCC 22931. Similarly, S. typhimurium isolates were compared with the standard S. typhimurium strain ATCC 14028.


Subject(s)
Amidohydrolases/genetics , Bacterial Proteins/genetics , Campylobacter jejuni/genetics , Food Microbiology , Polymerase Chain Reaction , Poultry Products/microbiology , Ribotyping , Salmonella typhimurium/genetics , Virulence Factors/genetics , Animals , Anti-Bacterial Agents/pharmacology , Campylobacter Infections/microbiology , Campylobacter jejuni/drug effects , Campylobacter jejuni/isolation & purification , Drug Resistance, Multiple, Bacterial/genetics , Food Handling , Microbial Sensitivity Tests , Salmonella Food Poisoning/microbiology , Salmonella typhimurium/drug effects , Salmonella typhimurium/isolation & purification , Saudi Arabia
7.
Int J Mol Sci ; 22(16)2021 Aug 18.
Article in English | MEDLINE | ID: mdl-34445577

ABSTRACT

Infections by the zoonotic foodborne bacterium Campylobacter jejuni (C. jejuni) are among the most frequent causes of bacterial gastroenteritis worldwide. The aim was to evaluate the relationship between epithelial barrier disruption, mucosal immune activation, and vitamin D (VD) treatment during C. jejuni infection, using intestinal epithelial cells and mouse models focused on the interaction of C. jejuni with the VD signaling pathway and VD treatment to improve C. jejuni-induced barrier dysfunction. Our RNA-Seq data from campylobacteriosis patients demonstrate inhibition of VD receptor (VDR) downstream targets, consistent with suppression of immune function. Barrier-preserving effects of VD addition were identified in C. jejuni-infected epithelial cells and IL-10-/- mice. Furthermore, interference of C. jejuni with the VDR pathway was shown via VDR/retinoid X receptor (RXR) interaction. Paracellular leakiness of infected epithelia correlated with tight junction (TJ) protein redistribution off the TJ domain and apoptosis induction. Supplementation with VD reversed barrier impairment and prevented inhibition of the VDR pathway, as shown by restoration of transepithelial electrical resistance and fluorescein (332 Da) permeability. We conclude that VD treatment restores gut epithelial barrier functionality and decreases bacterial transmigration and might, therefore, be a promising compound for C. jejuni treatment in humans and animals.


Subject(s)
Campylobacter Infections/complications , Cell Membrane Permeability , Epithelial Cells/drug effects , Interleukin-10/physiology , Intestinal Mucosa/drug effects , Vitamin D/pharmacology , Animals , Campylobacter Infections/microbiology , Campylobacter jejuni/isolation & purification , Epithelial Cells/immunology , Epithelial Cells/metabolism , Epithelial Cells/pathology , Humans , Intestinal Mucosa/immunology , Intestinal Mucosa/metabolism , Intestinal Mucosa/pathology , Mice , Mice, Inbred C57BL , Mice, Knockout , Tight Junction Proteins/metabolism , Vitamins/pharmacology
8.
Sci Rep ; 11(1): 12738, 2021 06 17.
Article in English | MEDLINE | ID: mdl-34140589

ABSTRACT

Both Campylobacter- and Shigella-induced invasive enteritis are common in under-5 Bangladeshi children. Our study aimed to determine the factors associated with Campylobacter and Shigella enteritis among under-5 children, the post-infection worsening growth, and the household cost of invasive enteritis. Data of children having Shigella (591/803) and Campylobacter (246/1148) isolated from the fecal specimen in Bangladesh were extracted from the Global Enteric Multicenter Study (GEMS) for the period December 2007 to March 2011. In multiple logistic regression analysis, fever was observed more frequently among shigellosis cases [adjusted OR 2.21; (95% CI 1.58, 3.09)]. Breastfeeding [aOR 0.55; (95% CI 0.37, 0.81)] was found to be protective against Shigella. The generalized estimating equations multivariable model identified a negative association between Shigella and weight-for-height z score [aOR - 0.11; (95% CI - 0.21, - 0.001)]; a positive association between symptomatic Campylobacter and weight-for-age z score [aOR 0.22; (95% CI 0.06, 0.37)] and weight-for-height z score [aOR 0.22; (95% CI 0.08, 0.37)]. Total costs incurred by households were more in shigellosis children than Campylobacter-induced enteritis ($4.27 vs. $3.49). Households with low-level maternal education tended to incur less cost in case of their shigellosis children. Our findings underscore the need for preventive strategies targeting Shigella infection, which could potentially reduce the disease burden, associated household costs, and child growth faltering.


Subject(s)
Cost of Illness , Enteritis/epidemiology , Growth , Bangladesh/epidemiology , Campylobacter jejuni/isolation & purification , Child, Preschool , Enteritis/economics , Enteritis/microbiology , Feces/microbiology , Humans , Risk Factors , Severity of Illness Index , Shigella flexneri/isolation & purification , Shigella sonnei/isolation & purification
9.
PLoS One ; 16(6): e0253797, 2021.
Article in English | MEDLINE | ID: mdl-34166472

ABSTRACT

Campylobacter species are one of the most common causative agents of gastroenteritis worldwide. Resistance against quinolone and macrolide antimicrobials, the most commonly used therapeutic options, poses a serious risk for campylobacteriosis treatment. Owing to whole genome sequencing advancements for rapid detection of antimicrobial resistance mechanisms, phenotypic and genotypic resistance trends along the "farm-to-fork" continuum can be determined. Here, we examined the resistance trends in 111 Campylobacter isolates (90 C. jejuni and 21 C. coli) recovered from clinical samples, commercial broiler carcasses and dairy products in Cairo, Egypt. Multidrug resistance (MDR) was observed in 10% of the isolates, mostly from C. coli. The prevalence of MDR was the highest in isolates collected from broiler carcasses (13.3%), followed by clinical isolates (10.5%), and finally isolates from dairy products (4%). The highest proportion of antimicrobial resistance in both species was against quinolones (ciprofloxacin and/or nalidixic acid) (68.4%), followed by tetracycline (51.3%), then erythromycin (12.6%) and aminoglycosides (streptomycin and/or gentamicin) (5.4%). Similar resistance rates were observed for quinolones, tetracycline, and erythromycin among isolates recovered from broiler carcasses and clinical samples highlighting the contribution of food of animal sources to human illness. Significant associations between phenotypic resistance and putative gene mutations was observed, with a high prevalence of the gyrA T86I substitution among quinolone resistant isolates, tet(O), tet(W), and tet(32) among tetracycline resistant isolates, and 23S rRNA A2075G and A2074T mutations among erythromycin resistant isolates. Emergence of resistance was attributed to the dissemination of resistance genes among various lineages, with the dominance of distinctive clones. For example, sub-lineages of CC828 in C. coli and CC21 in C. jejuni and the genetically related clonal complexes 'CC206 and CC48' and 'CC464, CC353, CC354, CC574', respectively, propagated across different niches sharing semi-homogenous resistance patterns.


Subject(s)
Bacterial Proteins/genetics , Campylobacter coli , Campylobacter jejuni , Chickens/microbiology , Dairy Products/microbiology , Drug Resistance, Bacterial/genetics , Mutation , Animals , Anti-Bacterial Agents/pharmacology , Campylobacter coli/genetics , Campylobacter coli/isolation & purification , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Farms , Food Microbiology , Humans , Microbial Sensitivity Tests
10.
Int J Food Microbiol ; 351: 109263, 2021 Aug 02.
Article in English | MEDLINE | ID: mdl-34116344

ABSTRACT

Campylobacter is the leading cause of foodborne human diarrhea worldwide. This microbe in the viable but non-culturable (VBNC) state can evade detection by routinely used culture-based methods and remain viable for extended periods of time. Bacteria in this dormancy state can resume their metabolic activity and virulence by resuscitation under favorable conditions, and subsequently cause infections. In this study, an assay combining loop-mediated isothermal amplification (LAMP) and propidium monoazide (PMA) treatment was developed for the detection and quantification of VBNC C. jejuni in agri-foods. PMA-qLAMP targeting the hipO gene demonstrated 100% high specificity to C. jejuni. A linear detection of C. jejuni was achieved between 8.77 × 102 and 8.77 × 07 CFU/mL with a coefficient of determination (R2) of 0.9956, indicating a good quantitative capacity. C. jejuni was effectively induced into the VBNC state by osmotic stress (i.e., 7% NaCl, w/v) over 48 h. VBNC C. jejuni cells were spiked into three representative food products and determined by PMA-qLAMP coupled with plating assay. The detection limits of PMA-qLAMP were 1.58 × 102 CFU/mL in milk, 3.78 × 102 CFU/g in chicken breast meat, and 4.33 × 102 CFU/g in romaine lettuce. PMA-qLAMP demonstrated rapid (25-40 min), specific (100% inclusivity and 100% exclusivity) and sensitive (~102 CFU/mL) determination of VBNC C. jejuni. This method can be applied in the agri-food industry to decrease the risks related to the consumption of contaminated agri-foods with pathogenic bacteria in the VBNC state and reduce the burden of C. jejuni infections to public health.


Subject(s)
Campylobacter jejuni/isolation & purification , Food Microbiology/methods , Animals , Azides , Campylobacter jejuni/genetics , Chickens , Genes, Bacterial/genetics , Intercalating Agents , Lactuca/microbiology , Milk/microbiology , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , Poultry/microbiology , Propidium/analogs & derivatives , Sensitivity and Specificity
11.
Sci Rep ; 11(1): 8998, 2021 04 26.
Article in English | MEDLINE | ID: mdl-33903652

ABSTRACT

Campylobacter, a leading cause of gastroenteritis in humans, asymptomatically colonises the intestinal tract of a wide range of animals.Although antimicrobial treatment is restricted to severe cases, the increase of antimicrobial resistance (AMR) is a concern. Considering the significant contribution of ruminants as reservoirs of resistant Campylobacter, Illumina whole-genome sequencing was used to characterise the mechanisms of AMR in Campylobacter jejuni and Campylobacter coli recovered from beef cattle, dairy cattle, and sheep in northern Spain. Genome analysis showed extensive genetic diversity that clearly separated both species. Resistance genotypes were identified by screening assembled sequences with BLASTn and ABRicate, and additional sequence alignments were performed to search for frameshift mutations and gene modifications. A high correlation was observed between phenotypic resistance to a given antimicrobial and the presence of the corresponding known resistance genes. Detailed sequence analysis allowed us to detect the recently described mosaic tet(O/M/O) gene in one C. coli, describe possible new alleles of blaOXA-61-like genes, and decipher the genetic context of aminoglycoside resistance genes, as well as the plasmid/chromosomal location of the different AMR genes and their implication for resistance spread. Updated resistance gene databases and detailed analysis of the matched open reading frames are needed to avoid errors when using WGS-based analysis pipelines for AMR detection in the absence of phenotypic data.


Subject(s)
Campylobacter coli , Campylobacter jejuni , Drug Resistance, Bacterial/genetics , Genetic Variation , Genome, Bacterial , Animals , Campylobacter coli/genetics , Campylobacter coli/isolation & purification , Campylobacter coli/metabolism , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Campylobacter jejuni/metabolism , Cattle , Genotype , Sheep , Whole Genome Sequencing
12.
PLoS One ; 16(3): e0248464, 2021.
Article in English | MEDLINE | ID: mdl-33755697

ABSTRACT

Very few reports describe all hospitalized patients with campylobacteriosis in the setting of a single waterborne outbreak. This study describes the demographics, comorbidities, clinical features, microbiology, treatment and complications of 67 hospitalized children and adults during a large waterborne outbreak of Campylobacter jejuni in Askoy, Norway in 2019, where more than 2000 people in a community became ill. We investigated factors that contributed to hospitalization and treatment choices. Data were collected from electronic patient records during and after the outbreak. Fifty adults and seventeen children were included with a biphasic age distribution peaking in toddlers and middle-aged adults. Most children, 14 out of 17, were below 4 years of age. Diarrhea was the most commonly reported symptom (99%), whereas few patients (9%) reported bloody stools. Comorbidities were frequent in adults (63%) and included cardiovascular disease, pre-existing gastrointestinal disease or chronic renal failure. Comorbidities in children (47%) were dominated by pulmonary and gastrointestinal diseases. Adult patients appeared more severely ill than children with longer duration of stay, higher levels of serum creatinine and CRP and rehydration therapy. Ninety-two percent of adult patients were treated with intravenous fluid as compared with 12% of children. Almost half of the admitted children received antibiotics. Two patients died, including a toddler. Both had significant complicating factors. The demographic and clinical findings presented may be useful for health care planning and patient management in Campylobacter outbreaks both in primary health care and in hospitals.


Subject(s)
Campylobacter Infections/epidemiology , Disease Outbreaks , Adolescent , Adult , Aged , Aged, 80 and over , Campylobacter jejuni/isolation & purification , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Norway/epidemiology , Young Adult
13.
Sci Rep ; 11(1): 5676, 2021 03 11.
Article in English | MEDLINE | ID: mdl-33707610

ABSTRACT

The sequencing, assembly, and analysis of bacterial genomes is central to tracking and characterizing foodborne pathogens. The bulk of bacterial genome sequencing at the US Food and Drug Administration is performed using short-read Illumina MiSeq technology, resulting in highly accurate but fragmented genomic sequences. The MinION sequencer from Oxford Nanopore is an evolving technology that produces long-read sequencing data with low equipment cost. The goal of this study was to compare Campylobacter genome assemblies generated from MiSeq and MinION data independently, as well as hybrid genome assemblies combining both data types. Two reference strains and two field isolates of C. jejuni were sequenced using MiSeq and MinION, and the sequence data were assembled using the software programs SPAdes and Canu, respectively. Hybrid genome assembly was performed using the program Unicycler. Comparison of the C. jejuni 81-176 and RM1221 genome assemblies to the PacBio reference genomes revealed that the SPAdes assemblies had the most accurate nucleotide identity, while the hybrid assemblies were the most contiguous. Assemblies generated only from MinION data using Canu were the least accurate, containing many indels and substitutions that affected downstream analyses. The hybrid sequencing approach was the most useful for detecting plasmids, large genome rearrangements, and repetitive elements such as rRNA and tRNA genes. The full genomes of both C. jejuni field isolates were completed and circularized using hybrid sequencing, and a plasmid was detected in one isolate. Continued development of nanopore sequencing technologies will likely enhance the accuracy of hybrid genome assemblies and enable public health laboratories to routinely generate complete circularized bacterial genome sequences.


Subject(s)
Campylobacter jejuni/genetics , Genome, Bacterial , High-Throughput Nucleotide Sequencing , Base Sequence , Campylobacter jejuni/isolation & purification , Molecular Sequence Annotation , Multilocus Sequence Typing , Reference Standards
14.
PLoS One ; 16(3): e0246843, 2021.
Article in English | MEDLINE | ID: mdl-33690659

ABSTRACT

Campylobacter spp. is a significant and prevalent public health hazard globally. Campylobacter jejuni is the most frequently recovered species from human cases and poultry are considered the most important reservoir for its transmission to humans. In this study, 30 Campylobacter jejuni isolates were selected from clinical (n = 15) and broiler (n = 15) sources from a larger cohort, based on source, virulence, and antimicrobial resistance profiles. The objective of this study was to further characterise the genomes of these isolates including MLST types, population structure, pan-genome, as well as virulence and antimicrobial resistance determinants. A total of 18 sequence types and 12 clonal complexes were identified. The most common clonal complex was ST-45, which was found in both clinical and broiler samples. We characterised the biological functions that were associated with the core and accessory genomes of the isolates in this study. No significant difference in the prevalence of virulence or antimicrobial resistance determinants was observed between clinical and broiler isolates, although genes associated with severe illness such as neuABC, wlaN and cstIII were only detected in clinical isolates. The ubiquity of virulence factors associated with motility, invasion and cytolethal distending toxin (CDT) synthesis in both clinical and broiler C. jejuni genomes and genetic similarities between groups of broiler and clinical C. jejuni reaffirm that C. jejuni from poultry remains a significant threat to public health.


Subject(s)
Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Poultry/microbiology , Animals , Campylobacter jejuni/pathogenicity , Humans , Ireland , Multilocus Sequence Typing , Virulence
15.
Can J Microbiol ; 67(4): 301-309, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33703923

ABSTRACT

The aims of this study were to evaluate the ability of Campylobacter jejuni isolated from a poultry slaughterhouse to form biofilm in the presence and absence of Pseudomonas aeruginosa, and the effect of surface (stainless steel, polystyrene), temperature (7, 25, and 42 °C), and oxygen concentration (microaerophilic and aerobic conditions) on the formation of biofilm. The genes ahpC, cadF, clpP, dnaJ, docA, flaA, flaB, katA, kpsM, luxS, racR, and sodB, related to biofilm formation by C. jejuni, were also investigated. All isolates formed biofilm on stainless steel and on polystyrene, in both aerobic and microaerophilic atmospheres, including temperatures not optimal for C. jejuni growth (7 and 25 °C), and biofilm also was formed in the presence of P. aeruginosa. In dual-species biofilm on stainless steel, biofilm formation was 2-6 log CFU·cm-2 higher at 7 °C for all isolates, in comparison with monospecies biofilm. Ten genes (ahpC, cadF, clpP, dnaJ, docA, flaA, flaB, luxS, racR, and sodB) were detected in all isolates, but katA and kpsM were found in four and six isolates, respectively. The results obtained are of concern because the poultry C. jejuni isolates form biofilm in different conditions, which is enhanced in the presence of other biofilm formers, such as P. aeruginosa.


Subject(s)
Biofilms/growth & development , Campylobacter jejuni/physiology , Poultry/microbiology , Pseudomonas aeruginosa/physiology , Abattoirs , Animals , Campylobacter jejuni/isolation & purification , Microbial Interactions , Oxygen/analysis , Surface Properties , Temperature
16.
Foodborne Pathog Dis ; 18(5): 322-330, 2021 05.
Article in English | MEDLINE | ID: mdl-33656913

ABSTRACT

Our study aimed to determine the prevalence of Campylobacter jejuni isolated from raw milk, cheese, and human stool samples in Beni-Suef Governorate, Egypt, and to characterize the antibiotic resistance profile and virulence genes of the isolates. An additional objective was to evaluate the effectiveness of cinnamon oil and Lactobacillus acidophilus La5 for controlling C. jejuni in cheese. A total of 200 samples of raw milk and dairy products, including 50 samples of raw milk and 150 samples of three different types of cheese were used. Fifty-three human stool samples were also collected. The samples were tested for the presence of C. jejuni using culture and molecular methods. Campylobacter spp. were isolated from 9.5% (19/200) of the raw milk and cheese samples. The highest prevalence was observed in milk samples (18%), followed by Kareish cheese (14%) and Talaga cheese (6%). In contrast, C. jejuni was not found in any of the Feta cheese samples. Of the human stool samples, 21 (39.6%) were positive for C. jejuni. Of the isolates, 60-90% were highly resistant to the antimicrobial agents tested, that is, nalidixic acid, ciprofloxacin, and tetracycline. Virulent cadF and cdtA genes were detected in all isolates. As milk and dairy products are important sources of contamination, reducing the level of C. jejuni in them will lower the risk to consumers. We showed that L. acidophilus La5 was able to control C. jejuni in Kareish cheese, but cinnamon oil was less effective.


Subject(s)
Campylobacter jejuni/isolation & purification , Cheese/microbiology , Feces/microbiology , Food Microbiology/statistics & numerical data , Milk/microbiology , Animals , Campylobacter jejuni/drug effects , Campylobacter jejuni/pathogenicity , Cinnamomum zeylanicum , Drug Resistance, Microbial , Egypt/epidemiology , Humans , Lactobacillus acidophilus , Oils, Volatile/pharmacology , Prevalence , Virulence Factors/isolation & purification
17.
PLoS One ; 16(2): e0246755, 2021.
Article in English | MEDLINE | ID: mdl-33566816

ABSTRACT

Campylobacter jejuni and Campylobacter coli are globally recognized as a major cause of bacterial foodborne gastroenteritis. A cross-sectional study was conducted from October 2015 to May 2016 in Mekelle city to isolate, identify, and estimate the prevalence of C. jejuni and C. coli in raw meat samples and to determine their antibiotic susceptibility pattern. A total of 384 raw meat samples were randomly collected from bovine (n = 210), goat (n = 108), and chicken (n = 66), and isolation and identification of Campylobacter spp. were performed using standard bacteriological techniques and PCR. Antibiotic susceptibility test was performed using disc diffusion method. Of the total 384 raw meat samples, 64 (16.67%) were found positive for Campylobacter spp. The highest prevalence of Campylobacter spp. was found in chicken meat (43.93%) followed by bovine meat (11.90%) and goat meat (9.25%). The most prevalent Campylobacter spp. isolated from meat samples was C. jejuni (81.25%). The overall prevalence of Campylobacter in restaurants, butcher shops, and abattoir was 43.93%, 18.30%, and 9.30%, respectively. 96.8%, 81.25%, 75%, and 71% of the Campylobacter spp. isolates were sensitive to norfloxacin, erythromycin, chloramphenicol, and sulphamethoxazole-trimethoprim, respectively. However, 96.9%, 85.9%, and 50% of the isolates were resistant to ampicillin, amoxicillin, and streptomycin, respectively. Strains that developed multi-drug resistant were 68.7%. The result of this study revealed the occurrence of Campylobacter in bovine, goat, and chicken meats. Hence, there is a chance of acquiring infection via consumption of raw or undercooked meat. Thus, implementation of hygienic practices from a slaughterhouse to the retailers, proper handling and cooking of foods of meat are very important in preventing Campylobacter infection.


Subject(s)
Campylobacter Infections/microbiology , Campylobacter coli/drug effects , Campylobacter jejuni/drug effects , Chickens/microbiology , Drug Resistance, Multiple, Bacterial , Goats/microbiology , Meat/microbiology , Amoxicillin/pharmacology , Ampicillin/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Campylobacter coli/isolation & purification , Campylobacter jejuni/isolation & purification , Cattle , Chloramphenicol/pharmacology , Cross-Sectional Studies , Erythromycin/pharmacology , Ethiopia , Microbial Sensitivity Tests , Norfloxacin/pharmacology , Prevalence , Streptomycin/pharmacology , Sulfamethoxazole/pharmacology , Trimethoprim/pharmacology
18.
Article in English | MEDLINE | ID: mdl-33547152

ABSTRACT

OBJECTIVE: To identify the clinical phenotypes and infectious triggers in the 2019 Peruvian Guillain-Barré syndrome (GBS) outbreak. METHODS: We prospectively collected clinical and neurophysiologic data of patients with GBS admitted to a tertiary hospital in Lima, Peru, between May and August 2019. Molecular, immunologic, and microbiological methods were used to identify causative infectious agents. Sera from 41 controls were compared with cases for antibodies to Campylobacter jejuni and gangliosides. Genomic analysis was performed on 4 C jejuni isolates. RESULTS: The 49 included patients had a median age of 44 years (interquartile range [IQR] 30-54 years), and 28 (57%) were male. Thirty-two (65%) had symptoms of a preceding infection: 24 (49%) diarrhea and 13 (27%) upper respiratory tract infection. The median time between infectious to neurologic symptoms was 3 days (IQR 2-9 days). Eighty percent had a pure motor form of GBS, 21 (43%) had the axonal electrophysiologic subtype, and 18% the demyelinating subtype. Evidence of recent C jejuni infection was found in 28/43 (65%). No evidence of recent arbovirus infection was found. Twenty-three cases vs 11 controls (OR 3.3, confidence interval [CI] 95% 1.2-9.2, p < 0.01) had IgM and/or IgA antibodies against C jejuni. Anti-GM1:phosphatidylserine and/or anti-GT1a:GM1 heteromeric complex antibodies were strongly positive in cases (92.9% sensitivity and 68.3% specificity). Genomic analysis showed that the C jejuni strains were closely related and had the Asn51 polymorphism at cstII gene. CONCLUSIONS: Our study indicates that the 2019 Peruvian GBS outbreak was associated with C jejuni infection and that the C jejuni strains linked to GBS circulate widely in different parts of the world.


Subject(s)
Campylobacter Infections/diagnosis , Campylobacter Infections/epidemiology , Campylobacter jejuni/isolation & purification , Disease Outbreaks , Guillain-Barre Syndrome/diagnosis , Guillain-Barre Syndrome/epidemiology , Adult , Campylobacter Infections/blood , Case-Control Studies , Female , Guillain-Barre Syndrome/blood , Humans , Male , Middle Aged , Peru/epidemiology
19.
PLoS One ; 16(2): e0246571, 2021.
Article in English | MEDLINE | ID: mdl-33571292

ABSTRACT

The Center for Disease Control and Prevention identifies antimicrobial resistant (AMR) Campylobacter as a serious threat to U.S. public health due to high community burden, increased transmissibility, and limited treatability. The National Antimicrobial Resistance Monitoring System (NARMS) plays an important role in surveillance of AMR bacterial pathogens in humans, food animals and retail meats. This study investigated C. coli and C. jejuni from live food animals, poultry carcasses at production, and retail meat in North Carolina between January 2018-December 2019. Whole genome sequencing and bioinformatics were used for phenotypic and genotypic characterization to compare AMR profiles, virulence factors associated with Guillain-Barré Syndrome (GBS) (neuABC and cst-II or cst-III), and phylogenic linkage between 541 Campylobacter isolates (C. coli n = 343, C. jejuni n = 198). Overall, 90.4% (489/541) Campylobacter isolates tested positive for AMR genes, while 43% (233/541) carried resistance genes for three or more antibiotic classes and were classified molecularly multidrug resistant. AMR gene frequencies were highest against tetracyclines (64.3%), beta-lactams (63.6%), aminoglycosides (38.6%), macrolides (34.8%), quinolones (24.4%), lincosamides (13.5%), and streptothricins (5%). A total of 57.6% (114/198) C. jejuni carried GBS virulence factors, while three C. coli carried the C. jejuni-like lipooligosaccharide locus, neuABC and cst-II. Further evidence of C. coli and C. jejuni interspecies genomic exchange was observed in identical multilocus sequence typing, shared sequence type (ST) 7818 clonal complex 828, and identical species-indicator genes mapA, ceuE, and hipO. There was a significant increase in novel STs from 2018 to 2019 (2 in 2018 and 21 in 2019, p<0.002), illustrating variable Campylobacter genomes within food animal production. Introgression between C. coli and C. jejuni may aid pathogen adaption, lead to higher AMR and increase Campylobacter persistence in food processing. Future studies should further characterize interspecies gene transfer and evolutionary trends in food animal production to track evolving risks to public health.


Subject(s)
Campylobacter coli/genetics , Campylobacter jejuni/genetics , Drug Resistance, Bacterial , Gene Transfer, Horizontal , Animals , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Campylobacter coli/drug effects , Campylobacter coli/isolation & purification , Campylobacter jejuni/drug effects , Campylobacter jejuni/isolation & purification , Meat/microbiology , North Carolina , Poultry/microbiology
20.
J Med Microbiol ; 70(3)2021 Mar.
Article in English | MEDLINE | ID: mdl-33475480

ABSTRACT

Introduction. In May-June 2018, an outbreak of campylobacteriosis involved students and school staff from kindergartens and primary schools in Pescara, southern Italy.Aim. We present details of the epidemiological and microbiological investigation, and the findings of the analytical study, as well as the implemented control measures.Methodology. To identify possible risk factors associated with the observed outbreak, a case control study was conducted using a questionnaire to collect information on the date of symptoms onset, type and duration of symptoms, type of healthcare contact, school attendance, and food items consumed at school lunches during the presumed days of exposure. Attack rates were calculated for each date and school. Logistic regression models were used to estimate the odds ratios of being a case and the odds of illness by food items consumed, respectively. Moreover, we carried out a comparative genomic analysis using whole genome multilocus sequence typing (wgMLST) of Campylobacter jejuni strains isolated during the outbreak investigation to identify the source of the outbreak.Results. Overall, 222 probable cases from 21 schools were identified, and C. jejuni was successfully isolated from 60 patients. The meals in the schools involved were provided by two cooking centres managed by a joint venture between two food companies. Environmental and food sampling, epidemiological and microbiological analyses, as well as a case control study with 176 cases and 62 controls from the same schools were performed to identify the source of the outbreak. The highest attack rate was recorded among those having lunch at school on 29 May (7.8 %), and the most likely exposure was 'caciotta' cheese (odds ratio 2.40, 95 % confidence interval 1.10-5.26, P=0.028). C. jejuni was isolated from the cheese, and wgMLST showed that the human and cheese isolates belonged to the same genomic cluster, confirming that the cheese was the vehicle of the infection.Conclusion. It is plausible that a failure of the pasteurization process contributed to the contamination of the cheese batches. Timely suspension of the catering service and summer closure of the schools prevented further spread.


Subject(s)
Campylobacter Infections , Campylobacter jejuni/isolation & purification , Cheese/microbiology , Disease Outbreaks , Food Microbiology , Foodborne Diseases/microbiology , Adult , Campylobacter Infections/epidemiology , Campylobacter Infections/microbiology , Case-Control Studies , Child , Child, Preschool , Female , Foodborne Diseases/epidemiology , Humans , Italy , Male , Pasteurization , Schools , Surveys and Questionnaires
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