Pathogenic Campylobacter Species Research Articles

Campylobacter infection of young children in Colombia and its impact on the gastrointestinal environment.

Campylobacter infections are a leading cause of bacterial-derived gastroenteritis worldwide with particularly profound impacts on pediatric patients in low- and middle-income countries. It remains unclear how Campylobacter impacts these hosts, though it is becoming increasingly evident that it is a multifactorial process that depends on the host immune response, the gastrointestinal microbiota, various bacterial factors, and host nutritional status. Since these factors likely vary between adult and pediatric patients in different regions of the world, it is important that studies define these attributes in well-characterized clinical cohorts in diverse settings. In this study, we analyzed the fecal microbiota and the metabolomic and micronutrient profiles of asymptomatic and symptomatic pediatric patients in Colombia who were either infected or uninfected with Campylobacter during a case-controlled study on acute diarrheal disease. Here, we report that the microbiome of Campylobacter-infected children only changed in their abundance of Campylobacter spp. despite the inclusion of children with or without diarrhea. In addition to increased Campylobacter, computational models were used to identify fecal metabolites that were associated with Campylobacter infection and found that glucose-6-phosphate and homovanillic acid were the strongest predictors of infection in these pediatric patients, which suggests that colonocyte metabolism is impacted during infection. Despite changes to the fecal metabolome, the concentrations of intestinal minerals and trace elements were not significantly impacted by Campylobacter infection but were elevated in uninfected children with diarrhea.IMPORTANCEGastrointestinal infection with pathogenic Campylobacter species has long been recognized as a significant cause of human morbidity. Recently, it has been observed that pediatric populations in low- and middle-income countries are uniquely impacted by these organisms in that infected children can be persistently colonized, develop enteric dysfunction, and exhibit reduced development and growth. While the association of Campylobacter species with these long-term effects continues to emerge, the impact of infection on the gastrointestinal environment of these children remains uncharacterized. To address this knowledge gap, our group leveraged clinical samples collected during a previous study on gastrointestinal infections in pediatric patients to examine the fecal microbiota, metabolome, and micronutrient profiles of those infected with Campylobacter species and found that the metabolome was impacted in a way that suggests gastrointestinal cell metabolism is affected during infection, which is some of the first data indicating how gastrointestinal health in these patients may be affected.

Prevalence of multidrug-resistant Campylobacter species in wastewater effluents: A menace of environmental and public health concern.

The prevalence of multidrug-resistant Campylobacter species in wastewater effluents presents a formidable challenge at the intersection of environmental sustainability and public health. This study examined the presence of multidrug-resistant Campylobacter in wastewater effluents in the Eastern Cape Province, South Africa, and its implications for environmental ecosystems and public health. Forty-five samples from household effluent (HHE) and wastewater treatment plant effluent (WWTPE) were collected at different geographical locations within the province between April and September 2022. The counts of the presumptive Campylobacter genus ranged from 5.2 × 103 to 6.03 × 104 CFU/mL for HHE and 4.93 × 103 to 1.04 × 104 CFU/mL for WWTPE. About 42.55% of the samples were positive for Campylobacter species. Five virulence determinants including the cadF and wlaN were detected in all the isolates; however, flgR (19.23%), ciaB, and ceuE (15.38%) were less prevalent. The antibiogram profiles of confirmed Campylobacter isolates revealed high resistance (>55%) against all tested antibiotics ranging from 55.77% (nalidixic acid) to 92.30% (erythromycin), and resistance against the other antibiotics followed the order ciprofloxacin (51.92%), azithromycin (50%), and levofloxacin (48.08%). On the contrary, gentamicin was sensitive against 61.54% of the isolates, followed by imipenem (57.69%) and streptomycin (51.92%). The WWTPE's antibiotic resistance index (ARI) was 0.19, lower than the permitted Krumperman threshold of 0.2; and HHE's ARIs were higher. The isolates' respective multiple antibiotic resistance indexes (MARI) varied between 0.08 and 1.00. Among the phenotypically resistant Campylobacter isolates examined, 21 resistance determinants encoding resistance against β-lactam, carbapenems, aminoglycosides, phenicol, quinolones, tetracyclines, and macrolides were detected, which explains the phenotypic resistance observed in the study. This study concludes that the wastewaters in the study areas are important reservoirs of multidrug-resistant and potentially pathogenic Campylobacter species, suggesting the need for proper treatment of the wastewaters to eliminate the organisms in the effluents before discharge the final effluent to the receiving watershed.

Clinico-bacteriological Study and Molecular Detection of Campylobacter in Childhood Diarrhoea: A Cross-sectional Study

Introduction: Campylobacter infections cause diarrhoeal diseases as frequently as Salmonella and Shigella infections. The prevalence of Campylobacter infection among children with acute diarrhoea in developing countries ranges from 5-35%. Diagnosing Campylobacter infections is challenging as the organism is difficult to isolate, grow, and identify. Currently, no best-practice clinical or public health laboratory guidelines exist for laboratory diagnosis of Campylobacter infections. Aim: To explore the clinical and bacterial aspects of childhood diarrhoea, emphasising the prevalence and molecular detection of Campylobacter. Materials and Methods: A hospital-based cross-sectional descriptive study was conducted with 55 stool samples of children under five with diarrhoea or dysentery at the Department of Microbiology, JSS Medical College, Mysuru, Karnataka, India, from October 2016 to September 2017. All stool samples were inoculated onto Campylobacter selective and non selective media with filtration and incubated in microaerophilic conditions. The culture isolates were identified by standard phenotypic tests. Molecular characterisation of Campylobacter was performed targeting the Campylobacter adhesion to fibronectin F (cadF) gene. The presence of a phylogenetically conserved 16S ribosomal Ribonucleic Acid (16SrRNA) domain was studied, followed by specific detection of pathogenic Campylobacter species. The Statistical Package for Social Sciences (SPSS) version 22.0 was used for statistical analysis. Descriptive statistics like percentage, mean, and Standard Deviation (SD) were applied. Results: Campylobacter was isolated by culture in one out of 55 stool samples. The isolate was confirmed to be Campylobacter jejuni by phenotypic tests. Campylobacter genus-level Polymerase Chain Reaction (PCR) was positive for 6 samples (10.9%). Six positive samples were subjected to species-level PCR; all were positive for C. jejuni. Out of 55 stool samples, two diarrheagenic Escherichia coli, two Shigella sonnei, one Shigella dysenteriae, and one Salmonella enterica serovar Typhi were also identified. Conclusion: Culture is insufficiently sensitive for diagnosing Campylobacter infection compared with nucleic acid-based diagnostics. Nucleic acid-based diagnostics offer increased sensitivity, can determine both the presence and burden of infection, and can distinguish between Campylobacter infections at the species level. Therefore, PCR is recommended, if feasible, as the preferred diagnostic modality for detecting Campylobacter infection.

The potential role of migratory birds in the transmission of pathogenic Campylobacter species to broiler chickens in broiler poultry farms and live bird markets

BackgroundCampylobacter species (spp.) are one of the most important zoonotic bacteria possessing potential hazards for animal and human health worldwide. Migratory birds are implicated as significant carriers for microbes and a play very important role in the dissemination of Campylobacter to broiler chickens and their environment. The purpose of this investigation was to detect the prevalence, antibiotic resistant patterns, virulence and diversity of pathogenic Campylobacter spp. in 7 migratory bird species (Northern shoveler, Common pochard, Common teal, Northern pintail, Eared Grebe, Great Crested Grebe and Garganey) and broiler chickens that were collected from broiler poultry farms and live bird markets.ResultsThe prevalence of Campylobacter was 12.5% (25/200), of which 15% (15/100) was recovered from 5 migratory bird species only and 10% (10/100) from broiler chickens. At the level of migratory birds, eight isolates (53.3%) were Campylobacter jejuni (C. jejuni) and 7 isolates (46.7%) were Campylobacter coli (C. coli) meanwhile, in broiler chickens C. jejuni and C. coli were 50% (5/10) for each. All isolated strains had phenotypic resistance to doxycycline, while all of the isolates were susceptible to amikacin. The multidrug resistance to three, four or five antimicrobial classes was found in 72% (18/25) of the isolated strains. The multiantibiotic resistance index between the examined isolates was 0.22–0.77, with 10 antibiotic resistance patterns. The virulence of isolated Campylobacter strains (from both migratory birds and broiler chicken birds) was detected by targeting the VirB11, ciaB and iam genes which were recorded at 16%, 52% and 100%, respectively. Additionally, 100% and 84% of the antibiotic resistance genes were identified as tetA and BlaOXA-61, respectively.ConclusionsThe results of this study revealed the diversity between all the isolated strains from migratory birds and their similarity to broiler chicken isolates. The findings of the present study highlight the impact of migratory birds visiting Egypt and other countries on pathogenic Campylobacter spp. carrying pathogenic virulence and resistance genes, necessitating the application of biosecurity measures to prevent migratory birds from entering farms during their migration period.

Wild boars as reservoir for Campylobacter and Arcobacter

Campylobacteriosis is a significant public health concern with Campylobacter jejuni and Campylobacter coli as main causative agents. Moreover, there is an increasing recognition of other pathogenic Campylobacter species and Campylobacter-like organisms as Arcobacter. However, current knowledge on presence of Arcobacter species in wild boars (Sus scrofa) is lacking, and knowledge on Campylobacter species is based on methods favoring growth of thermotolerant species. In this study, fecal samples originating from 76 wild boars hunted in Campania region (Italy) were examined for the presence of Campylobacter(-like) organisms by a culture dependent approach. Three isolation protocols were performed in parallel: Arcobacter-selective agar plates, mCCDA plates and isolation by passive filtration onto non-selective blood agar plates were used as quantitative isolation methods. Enrichment broths, i.e. Arcobacter selective enrichment broth, Preston broth and CAT broth were used for qualitative detection of low levels or stressed Campylobacter(-like) organisms. The Arcobacter and Campylobacter isolates were identified at species level using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S ribosomal RNA (rRNA) sequence analysis. Overall, 41 (53.9%) of the animals excreted Arcobacter or Campylobacter while 38 (50.0%) shed Campylobacter and 8 (10.5%) Arcobacter. Campylobacter lanienae predominated and was isolated from 31 (40.8%) animals. No statistical difference between the age groups or gender with regard to the fecal excretion of Campylobacter(-like) organisms was observed. Thirty animals (39.5%) shed Campylobacter spp. exceeding levels of 10 ³ CFU g−1 feces. As samples were obtained from hunted wild boars intended for consumption, a potential contamination of meat with these bacterial pathogens must be considered.

A loop-mediated isothermal amplification (LAMP) assay for the consensus detection of human pathogenic Campylobacter species

Most rapid identification methods for Campylobacter are designed to detect thermotolerant Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli). A growing number of thermosensitive Campylobacter species are now gaining recognition as emerging human pathogens. Methods are lacking for the rapid screening of these emerging species. Loop-mediated Isothermal Amplification (LAMP) is a nucleic acid amplification method that allows for the rapid and cost-effective detection of bacteria. Degenerate primers against the 16S rRNA sequences for C. jejuni, C. coli, C. lari, C. upsaliensis, C. ureolyticus, C. fetus, C. gracilis, C. rectus, and C. concisus were designed. Isothermal amplification was conducted using ATCC reference strains at 68 °C for 30 min using WarmStart® Colorimetric LAMP reagents. Positive reactions were indicated by a color change from pink to yellow; specificity to Campylobacter was confirmed using a restriction enzyme digest (RsaI). The developed LAMP reaction was specific for the reference strains, which was confirmed against an exclusivity panel that consisted of other enteric pathogens, including E. coli, Salmonella, Shigella, Helicobacter, and Arcobacter. This method was also evaluated for the detection of C. jejuni, C. coli, and C. lari in primary enrichment media from artificially contaminated fresh spinach samples. The LAMP method provides an option to rapidly screen for the presence of pathogenic Campylobacter spp. in field surveillance and trace-back analysis.

Culture Methods to Determine the Limit of Detection and Survival in Transport Media of Campylobacter Jejuni in Human Fecal Specimens.

A culture from human stool for diagnosis of Campylobacter-based intestinal illness takes several days, a wait that taxes the fortitude of the physician and the patient. A culture is also prone to false negative results from random loss of viability during specimen handling, overgrowth of other fecal flora, and poor growth of several pathogenic Campylobacter species on traditional media. These problems can confound clinical decisions on patient treatment and have limited the field from answering fundamental questions on Campylobacter growth and infections. We describe a procedure that estimates the lower limit of bacterial numbers that can be detected by a culture and a method for quantifying survival of C. jejuni in media used for transport of this fragile organism. Knowing this information, it becomes possible to set clinically relevant detection thresholds for diagnostic tests and address unstudied issues of whether non-symptomatic colonization is prevalent, if co-infection with other enteric pathogens is common, or if bacterial load correlates with symptoms or serious sequelae. The study also included testing of 1,552 prospectively collected patient diarrheal fecal specimens that were initially classified by conventional culture and further tested by a new enzyme immunoassay. Positive and discrepant specimens were then screened by four molecular methods to assign true-positive or true-negative status. The 5 non-culture methods showed complete agreement on all 48 positive and discrepant specimens, while the culture mis-identified 14 (28%). The specimens that were incorrectly identified by culture included 13 false negative and 1 false positive sample. This basic protocol can be used with multiple Campylobacter spp. and will allow the numbers of Campylobacter bacteria that produce symptoms of gastroenteritis in humans to be determined and for prevalence rates to be updated.

Enhanced microbiological surveillance reveals that temporal case clusters contribute to the high rates of campylobacteriosis in a model agroecosystem

Infections by pathogenic Campylobacter species were determined in diarrheic (n = 2,217) and non-diarrheic control (n = 104) people in Southwestern Alberta (SWA), Canada over a 1-year period using specialized and conventional isolation, and direct PCR. Overall, 9.9% of diarrheic individuals were positive for C. jejuni (9.1%), C. upsaliensis (0.6%), and C. coli (0.5%). No C. lari was detected. Four diarrheic individuals were co-infected with C. jejuni and C. coli, and four different individuals were co-infected with C. jejuni and C. upsaliensis. Two control individuals were positive for C. jejuni. Approximately 50% of stools containing C. jejuni and/or C. coli were deemed negative by conventional isolation. Direct PCR for C. jejuni was less effective than culture-based detection. Most C. jejuni infections occurred in people living in the urban centers, but the prevalence of the bacterium was lower in females than males living in urban locations, and both males and females living in rural locations. Although C. jejuni was detected throughout the year, a trend for higher infection rates was observed in the late spring to early fall with a peak in August. Forty-six C. jejuni subtype clusters were identified, including 44 temporal case clusters attributed to 28 subtype groupings. The majority of infections (70.3%) were linked to subtypes associated with beef cattle. We conclude that many occurrences of pathogenic Campylobacter species were not detected by the conventional laboratory methodology, and temporal case clusters of C. jejuni subtypes associated with cattle contribute to the high rates of campylobacteriosis in SWA.

Prevalence of most common human pathogenic Campylobacter spp. in dogs and cats in Styria, Austria.

The aim of this study was to determine the frequency of occurrence of most common human pathogenic Campylobacter species, Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli), in dogs and cats in Styria, Austria. In the period from April 2010 to April 2012, 842 faecal samples from dogs and cats from Styria, Austria were examined for Campylobacter (C.) species (spp.). All samples were subjected to qualitative microbiological culture testing, and additionally, some of them have been studied using qualitative real‐time PCR. In microbiological culture, 5.9% of all samples investigated were C. spp. positive. With 3.1% out of positive samples, C. jejuni was the most common type. Campylobacter upsaliensis (C. upsaliensis) was detected only in 0.5% of the samples. The remaining positive samples (2.4%) were classified as C. species (sp.). C. coli could not be found in any of the samples. A higher prevalence of C. jejuni was found in kittens with 14.3% and in diarrhoeic dogs (7.4%) and cats (23.8%). The real‐time PCR revealed for dogs and cats together, 27% of C. jejuni‐positive faecal and 8% positive faecal swap samples. The obtained C. jejuni strains underwent antibiotic resistance testing using three different tests (agar diffusion, MIC testing and E‐test) with different numbers of antibiotics. From the antibiotics used in this study, several showed high test‐dependent resistance rates (cephalexin, cefovecin, kanamycin, sulfamethoxazole/trimethoprim, penicillin G, ciprofloxacin, enrofloxacin, marbofloxacin, nalidixic acid). Overall, the prevalence of C. spp. in this study was very low compared to others, with the exception of C. jejuni in kittens and diarrhoeic animals. The results of the real‐time PCR suggest that the rate of colonization of C. jejuni was actually higher than the results of the culture showed. As the resistance rates of C. jejuni isolates partly were very high, possible transmission of (multi‐) resistant C. jejuni strains to humans especially from kittens and diarrhoeic animals must be expected.

Campylobacter QMRA: A Bayesian Estimation of Prevalence and Concentration in Retail Foods Under Clustering and Heavy Censoring.

A Bayesian statistical temporal-prevalence-concentration model (TPCM) was built to assess the prevalence and concentration of pathogenic campylobacter species in batches of fresh chicken and turkey meat at retail. The data set was collected from Finnish grocery stores in all the seasons of the year. Observations at low concentration levels are often censored due to the limit of determination of the microbiological methods. This model utilized the potential of Bayesian methods to borrow strength from related samples in order to perform under heavy censoring. In this extreme case the majority of the observed batch-specific concentrations was below the limit of determination. The hierarchical structure was included in the model in order to take into account the within-batch and between-batch variability, which may have a significant impact on the sample outcome depending on the sampling plan. Temporal changes in the prevalence of campylobacter were modeled using a Markovian time series. The proposed model is adaptable for other pathogens if the same type of data set is available. The computation of the model was performed using OpenBUGS software.

A molecular (PCR) survey on abortions caused by Campylobacter spp. in sheep flocks located on the suburb of Tabriz

Campylobacteriosis is an important infectious disease of animals and humans caused by the pathogenic Campylobacter species. A total number of 132 aborted sheep fetuses and related placentas were admitted to the large animal clinic at the University of Tabriz, from October 2010 to March 2011. Tissue samples were collected from several fetal organs including liver, brain, kidney, lung, spleen, heart, stomach fluid and placenta, then separately pulverized under liquid nitrogen and finally stored at -20°C until DNA extraction. Of 132 submissions (fetuses and placentas), 12 (9.09%) and 2(1.51%) samples were diagnosed positive to the Campylobacter fetus subsp. fetus and Campylobacter jejuni by the PCR protocol, respectively. No samples were positive for Campylobacter coli.

Potentially pathogenic Campylobacter species among farm animals in rural areas of Limpopo province, South Africa: A case study of chickens and cattles

This study investigated the prevalence of thermophilic pathogenic Campylobacter in farm settlements in South Africa. Six hundred freshly voided faeces samples were collected from chicken and cattle (300 each) and analyzed on mCCDA supplemented with campylobacter supplement and incubated at 42°C. Out of this, 106 (35.3%) of the chicken faeces were positive for Campylobacter, 58 (19.3%) from cattle faeces were positive for Campylobacter. Ninety (84.9%) isolates from chicken were C. jejuni, while 16 (15.1%) were C. coli. Of the 58 isolates from cattle, 42 (72.4%) were C. jejuni, while 16 (26.7%) were C. coli. C. jejuni was more prevalent in chicken and cattle. The prevalence was higher in chicken than cattle, the prevalence was statistically significant at P<0.005. Diarrhea faeces from chicken and cattle contain more C. jejuni than C. coli. The prevalence skewed more to chicken than cattle. Different levels of resistance were noted amongst isolates from chicken and cattle. Study of prevalence of resistance to ciprofloxacin (a fluoroquinolone) showed that C. coli from cattle were more resistant to this antibiotic (C. jejuni, 33.3%; C. coli, 56.3%); from chicken (C. jejuni, 29%; C. coli, 37.5%). Varied resistance was shown to other antibiotics by the isolates. The resistance by isolates to ciprofloxacin, a fluoroquinolone is worrisome since it is used as therapeutic agent against campylobacteriosis. Of more concern is the multiple resistances shown by these isolates to the applied antimicrobials as resistance genes can be transferred to other microbes in the environment horizontally. Key word: Prevalence, resistance, faeces, significant, campylobacteriosis, thermophilic.

Comparison between the biofilm initiation ofCampylobacter jejuniandCampylobacter colistrains to an inert surface using BioFilm Ring Test®

The adhesion to an inert surface (the first step of biofilm formation) of the two main pathogenic Campylobacter species, Campylobacter jejuni and Campylobacter coli, isolated from diverse origins, was compared. Adhesion assays were conducted in 96-well, polystyrene microtiter plates using the BioFilm Ring Test method. This new technique, based on magnetic bead entrapment, was shown to be suitable for analysing the adhesion of Campylobacter sp. strains by comparing the adhesion of four C. jejuni strains as revealed by the BioFilm Ring Test and immunodetection. Among the 46 strains tested, C. jejuni and C. coli displayed different adhesion capabilities ranging from no adhesion to strong adhesion. However, no strain of C. coli was strongly adherent, and statistically, C. coli adhered less to an inert surface than C. jejuni. In addition, strains isolated from animals or carcasses were less adherent than those isolated from food-processing and clinical cases. These observations suggest that the food environment and the human body could have selected strains with greater adhesion. The adhesion capability of strains could partly explain the cross-contamination or re-contamination of food products by Campylobacter. This property could provide a mode of survival for Campylobacter in the food chain.

A Multiplex Polymerase Chain Reaction for the Differentiation of Campylobacter jejuni and Campylobacter coli from a Swine Processing Facility and Characterization of Isolates by Pulsed-Field Gel Electrophoresis and Antibiotic Resistance Profiles

A multiplex polymerase chain reaction (PCR) was developed for the detection and speciation of 60 Campylobacter strains isolated from porcine rectal swabs and from different areas in a pork processing plant. The PCR assay was based on primers specific for the cadF gene of pathogenic Campylobacter species, a specific but undefined gene of Campylobacter jejuni, and the ceuE gene of Campylobacter coli. Further characterization of these isolates was established by pulsed-field gel electrophoresis (PFGE) analyses with the restriction endonuclease SmaI. In addition to molecular discrimination, the antibiotic resistance profiles of the isolates were examined by the Kirby Bauer disc diffusion method with 22 antibiotics. Differentiation of isolates by multiplex PCR identified 86.9% (52 of 60) as C. coli and 13.1% (8 of 60) as C. jejuni. Using the Molecular Analyst software, 60 PFGE types were identified. The percentages of relatedness among C. jejuni strains with PFGE ranged from 25 to 86%, while those among C. coli strains ranged from 34 to 99%. Among the 60 PFGE types, each of 12 C. coli isolates showed ≥90% similarity to one other isolate. The antibiotic resistance profiles of all 60 isolates were distinct. Analyses of antibiotic resistance profiles showed that all isolates were resistant to five or more antibiotics. Twenty-five percent (2 of 8) of C. jejuni isolates and 15% (8 of 52) of C. coli isolates were resistant to at least one of the three fluoroquinolones tested, antibiotics that are commonly used in the treatment of human Campylobacter infections. Three percent (2 of 60) of Campylobacter isolates examined were resistant to all three fluoroquinolones. On the basis of the PFGE and antibiotic resistance profiles, each of the 60 isolates was distinct, suggesting that C. jejuni and C. coli strains originating from diverse sources were present in porcine samples and in the pork processing plant.

Detection of pathogenic Campylobacter species in blood culture systems.

Because differences in recognition of Campylobacter fetus and Campylobacter jejuni in systemic infections may be due partially to differences in the ability to cultivate these organisms, we studied their growth characteristics in two widely used blood culture systems. In the Roche Septi-Chek system (Hoffman-La Roche, Inc., Nutley, N.J.), over a broad range of inocula all strains were detected in broth within 2 days and on paddles within 3 days. In the BACTEC 6B aerobic bottles (Johnston Laboratories, Inc., Towson, Md.), C. jejuni and C. fetus took a median of 5 and 3 days, respectively, to reach the growth index threshold. However, in the BACTEC 7D anaerobic bottles, C. fetus required a median of 2 days to reach the growth index threshold, whereas for C. jejuni the median was greater than 10 days. The poor performance of C. jejuni in both BACTEC systems may have been due to unfavorable incubation atmospheres and may partially explain why C. jejuni bacteremia is so infrequently detected. Overall, the Roche Septi-Chek system was excellent for detecting Campylobacter strains in blood cultures.