Campylobacter Research Articles

Unravelling the reservoirs for colonisation of infants with Campylobacter spp. in rural Ethiopia: protocol for a longitudinal study during a global pandemic and political tensions

IntroductionUndernutrition is an underlying cause of mortality in children under five (CU5) years of age. Animal-source foods have been shown to decrease malnutrition in CU5. Livestock are important reservoirs for...

Postinfective bowel dysfunction following Campylobacter enteritis is characterised by reduced microbiota diversity and impaired microbiota recovery

ObjectivesPersistent bowel dysfunction following gastroenteritis (postinfectious (PI)-BD) is well recognised, but the associated changes in microbiota remain unclear. Our aim was to define these changes after gastroenteritis caused by a...

Effect of Probiotic E. coli Nissle 1917 Supplementation on the Growth Performance, Immune Responses, Intestinal Morphology, and Gut Microbes of Campylobacter jejuni Infected Chickens.

ABSTRACTCampylobacter jejuni is the most common cause of bacterial foodborne gastroenteritis and holds significant public health importance. The continuing increase of antibiotic-resistant Campylobacter necessitates the development of antibiotic-alternative approaches to control infections in poultry and in humans. Here, we assessed the ability of E. coli Nissle 1917 (EcN; free and chitosan-alginate microencapsulated) to reduce C. jejuni colonization in chickens and measured the effect of EcN on the immune responses, intestinal morphology, and gut microbes of chickens. Our results showed that the supplementation of 3-week-old chickens daily with free EcN in drinking water resulted in a 2.0 log reduction of C. jejuni colonization in the cecum, whereas supplementing EcN orally three times a week, either free or microencapsulated, resulted in 2.0 and 2.5 log reductions of C. jejuni colonization, respectively. Gavaged free and microencapsulated EcN did not have an impact on the evenness or the richness of the cecal microbiota, but it did increase the villous height (VH), crypt depth (CD), and VH:CD ratio in the jejunum and ileum of chickens. Further, the supplementation of EcN (all types) increased C. jejuni-specific and total IgA and IgY antibodies in chicken’s serum. Microencapsulated EcN induced the expression of several cytokines and chemokines (1.6 to 4.3-fold), which activate the Th1, Th2, and Th17 pathways. Overall, microencapsulated EcN displayed promising effects as a potential nonantibiotic strategy to control C. jejuni colonization in chickens. Future studies on testing microencapsulated EcN in the feed and water of chickens raised on built-up floor litter would facilitate the development of EcN for industrial applications to control Campylobacter infections in poultry.

The impact of farm animal exposures on Campylobacter jejuni antibody levels in rural agricultural communities of Iowa, USA

BACKGROUND AND AIM: Campylobacter infections transmitted primarily through improperly cooked meat are the leading bacterial cause of diarrhea worldwide. How exposures to farm animals and animal waste (e.g., through occupational contact or residential proximity to animal feeding operations [AFOs]) contribute to infections has been relatively understudied. METHODS: We measured antibodies against Campylobacter jejuni using an enzyme-linked immunosorbent assay in participants from Iowa in the Biomarkers of Exposure and Effect in Agriculture study (n=304 farmers; n=146 non-farming controls from similar communities). Participants self-reported occupational animal contact within the past year. We used a statewide database of permitted AFOs that included animal counts standardized by animal size and manure production to characterize the intensity of AFO exposure within 5km of participants’ residences. We performed linear regressions of log-transformed optical density units (ODs) for immunoglobulin (Ig) A, G, and M antibody levels in relation to occupational animal and low/high AFO exposures, using non-farming controls with low AFO exposure (n=96) as the referent group. We adjusted for age, season, and other potential confounders. We further examined antibody levels by occupational contact with specific animal types. RESULTS: Occupational animal contact was associated with elevated IgA and IgG OD levels in those with low AFO exposure (geometric mean ratio [95% confidence interval]; IgA:1.34[1.07-1.67]; IgG:1.34[1.13-1.57]) as well as those with high AFO exposure (IgA:1.48[1.12-1.95]; IgG:1.34[1.09-1.65]). Antibody levels were higher in farmers raising poultry (IgA:2.20[1.03-4.71]; IgG:2.11[1.35-3.32]), swine (IgA:1.52[1.04-2.22]; IgG:1.50[1.17-1.94]), and dairy cattle (IgG:1.65[1.18-2.31]), particularly for IgG levels in those who reported spending time in poultry (2.29[1.25-4.19]) or swine (1.63[1.25-2.12]) confinements within the past month. We found no significant relationships between occupational animal contact and IgM or AFO exposure alone and any antibody levels. CONCLUSIONS: Occupational animal contact may be a stronger determinant of C. jejuni exposure than residential proximity to AFOs in agricultural communities. KEYWORDS: Campylobacter, animal exposures

Association between asymptomatic infections and linear growth in 18-24-month-old Malawian children.

Inadequate diet and frequent symptomatic infections are considered major causes of growth stunting in low-income countries, but interventions targeting these risk factors have achieved limited success. Asymptomatic infections can restrict growth, but little is known about their role in global stunting prevalence. We investigated factors related to length-for-age Z-score (LAZ) at 24 months by constructing an interconnected network of various infections, biomarkers of inflammation (as assessed by alpha-1-acid glycoprotein [AGP]), and growth (insulin-like growth factor 1[IGF-1] and collagen X biomarker [CXM]) at 18 months, as well as other children, maternal, and household level factors. Among 604 children, there was a continuous decline in mean LAZ and increased mean length deficit from birth to 24 months. At 18 monthsof age, the percentage of asymptomatic children who carried each pathogen was: 84.5% enterovirus, 15.5% parechovirus, 7.7% norovirus, 4.6% rhinovirus, 0.6% rotavirus, 69.6% Campylobacter, 53.8% Giardia lamblia, 11.9% malaria parasites, 10.2% Shigella, and 2.7% Cryptosporidium. The mean plasma IGF-1 concentration was 12.5 ng/ml and 68% of the children had systemic inflammation (plasma AGP concentration >1 g/L). Shigella infection was associated with lower LAZ at 24 months through both direct and indirect pathways, whereas enterovirus, norovirus, Campylobacter, Cryptosporidium, and malaria infections were associated with lower LAZ at 24 months indirectly, predominantly through increased systemic inflammation and reduced plasma IGF-1 and CXM concentration at 18 months.

In a warm climate, ventilation, indoor temperature and outdoor relative humidity have significant effects on Campylobacter spp. colonization in chicken broiler farms which can occur in only 2 days

A longitudinal study was conducted in five conventional broiler farms during a 2-year period to determine the dynamics of Campylobacter infection in a warm climate region (north-eastern Spain). Weekly sampling of 63 flocks was performed upon one-day-old chick placement, including animal and environmental samples. Campylobacter spp. detection was assessed by culture and non-culture methods. Environmental samples were also obtained from cleaned and disinfected houses prior to chick placement. Thirty-nine flocks (61.90%) became colonized during the growing period. First bird excreting Campylobacter was detected in 10-day-old chicks and the earliest a whole flock became positive was at 14 days of age, while the latest was at 39 days. Once Campylobacter was detected in chickens, the whole flock was colonized within 2–13 days. Campylobacter farm prevalence (positive flocks) ranged from 53.85% to 83.33% in four out of five farms, while the remaining farm showed a lower prevalence (38.5%). Logistic regression model showed that Campylobacter infection was more likely under higher minimal indoor temperature as well as at higher minimal outdoor relative humidity, characteristic of warm climates such as those from Mediterranean countries. Ventilation type was also significant (P = 0.021). No clear farm effect or seasonality were observed. Biosecurity improvements, specially at house level, are needed in broiler farms to prevent flock colonization and reduce the current high flock prevalence.

Quantitative Assessment of First Nations Drinking Water Distribution Systems for Detection and Prevalence of Thermophilic Campylobacter Species.

Water is considered a major route for transmitting human-associated pathogens. Although microbial water quality indicators are used to test for the presence of waterborne pathogens in drinking water, the two are poorly correlated. The current study investigates the prevalence of thermophilic DNA markers specific for Campylobacter spp. (C. jejuni and C. coli) in source water and throughout the water distribution systems of two First Nations communities in Manitoba, Canada. A total of 220 water samples were collected from various points of the drinking water distribution system (DWDS) between 2016 and 2018. Target Campylobacter spp. were always (100%) detected in a home with a fiberglass (CF) cistern, as well as the community standpipe (SP). The target bacteria were also frequently detected in treated water at the Water Treatment Plant (WTP) (78%), homes with polyethylene (CP) (60%) and concrete (CC) (58%) cisterns, homes with piped (P) water (43%) and water truck (T) samples (20%), with a maximum concentration of 1.9 × 103 cells 100 mL−1 (C. jejuni) and 5.6 × 105 cells 100 mL−1 (C. coli). Similarly, target bacteria were detected in 68% of the source water samples with a maximum concentration of 4.9 × 103 cells 100 mL−1 (C. jejuni) and 8.4 × 105 cells 100 mL−1 (C. coli). Neither target Campylobacter spp. was significantly associated with free and total chlorine concentrations in water. The study results indicate that there is an immediate need to monitor Campylobacter spp. in small communities of Canada and, particularly, to improve the DWDS in First Nations communities to minimize the risk of Campylobacter infection from drinking water sources. Further research is warranted in improving/developing processes and technologies to eliminate microbial contaminants from water.

Campylobacter jejuni subdural hygroma infection in a 2-year old boy: case report and a brief literature review

BackgroundCampylobacter jejuni is a common cause of acute gastroenteritis, but central nervous system infections are rare manifestations of Campylobacter infection. Therefore, C. jejuni trauma-related subdural hygroma infection in children is poorly described in the literature.Case presentationWe described a 2-year old boy with lobar holoprosencephaly presenting with subdural hygroma following head trauma. C. jejuni infection was confirmed from a subdural hygroma sample by culture as well as by DNA sequencing of a broad range 16S rDNA PCR product. Cerebrospinal fluid from the ventriculoperitoneal shunt remained sterile. Combined neurosurgical and antimicrobial treatment led to complete recovery. Review of the literature showed that the most common manifestation of Campylobacter central nervous system infection is meningitis, mostly in neonates, and subdural hygroma infection was described for only one case.ConclusionsSubdural hygroma infection caused by C. jejuni is a rare clinical condition in children. Molecular methods represent an important tool for the detection of rare or unexpected pathogens. No standard recommendations for antimicrobial treatment of C. jejuni subdural space infection in children are available, but meropenem treatment combined with surgery seems to be an effective approach.

Characterization of Campylobacter spp. isolated from chicken and at-risk people (children) in Cameroon: A review

Campylobacteriosis is a disease caused by the bacteria called Campylobacter spp, it is considered a public health problem. Campylobacter spp has so far been identified as the pathogen most responsible for bacterial gastroenteritis. Emphasis was placed on controlling the foodborne route of exposure. The assessment of the actual burden of Campylobacter in the African context, particularly in Cameroon, is hampered by the lack of reporting of diarrheal incidents and the ineffectiveness of monitoring and surveillance programs for foodborne illnesses, as well as the lack of attention given to Campylobacter as a causative agent of diarrhea for the sole benefit of Salmonella. This article aims to report on the characterization of Campylobacter spp in infected chickens and children in Central Africa, more specifically in Cameroon. Campylobacter infection is more prevalent in the pediatric population and has been isolated from farm animals, particularly chickens and foods of animal origin. The prevalence of Campylobacter in children with diarrhea under five years of age ranges from 15% to 23% in Angola. In chickens, the prevalence varies from 90% in Cameroon to 41.2% in Congo. This review also highlights the increased resistance of Campylobacter to common important antimicrobials, such as ciprofloxacin, tetracycline and erythromycin, in food animals and humans in Central Africa. The solution to limit the incidence in humans is to control and prevent the spread of pathogens in animals constituting the main reservoir of infections.

Loop-mediated isothermal amplification (LAMP) method for fast detection of Campylobacter spp in meat food products and environmental objects of a processing plant’s

There is constant necessity of developing the accurate and fast methods for detection of foodborne pathogens. Microorganisms of Campylobacter genus are one of the main causes of foodborne diseases worldwide. Fast identification of Campylobacter at all stages of the food life cycle is an efficient strategy to control foodborne campylobacteriosis. This article the authors evaluated a commercial loop-mediated isothermal amplification (LAMP) system with bioluminescence, called as the 3M™ Molecular Detection Analysis (MDA), which was used to find Campylobacter in food products with the help of a certain standard method, which is referred to as the reference method. The results of this study showed that the commercial LAMP based method is as efficient as the reference method, and features high specificity and minimum determinability (sensitivity). The LAMP based method has been shown to be a fast and reliable method for detection of Campylobacter spp. scarce presence (10 CFU/25 g) in meat, meat products, as well as carcass swabs and production facilities’ environment. The LAMP analysis required about 24–27 hours to achieve a result. However the LAMP based method will facilitate the detection of Campylobacter, as it provides much easier and faster detection of Cam pylobacter spp., including Campylobacter jejuni/Campylobacter coli, than standard microbiological methods. The LAMP based method is an efficient tool to prevent the spreading of Campylobacter spp. contamination in food products.

Quantitative microbial risk assessment as a tool for groundwater monitoring. A case study in the rural communities of Romania

Karst springs are an essential source of drinking water in rural communities in North-Western Romania. Due to the fractured dolomite aquifers, dolines, or blind valleys, these springs are vulnerable to contamination by human and zoonotic pathogens. In this study, we assessed the human health risk associated with the contamination of six springs’ water with gastrointestinal pathogens. The data on the occurrence of E. coli and estimated Campylobacter and Rotavirus for a year of monitoring from twenty-four samples were inputted into a Quantitative Microbial Risk Assessment (QMRA) model (Machdar et al. 2013). The QMRA was conducted following steps: hazard identification, dose-response assessment, exposure assessment, and risk characterization (Haas C N et al. 2014; World Health Organization 2016). In order to predict the total cases of illnesses for the spring water consumers, the probability of infection and illness was calculated for adults and children. Results have shown that the estimated probability of infection for enteropathogenic E. coli, Campylobacter, and Rotavirus ranged between 1.1 × 10-3 and 1.0 × 100, values much higher than the acceptable limit of 10-4 infections/person/year established by WHO (2016). The estimated probability of illness due to the pathogenic E. coli and Campylobacter infection was high, with values between 8.8 × 10-2 and 3.5 × 10-1 for five out of six groundwater sources. The estimated probability of illness due to the Rotavirus infection ranged between 4.0 × 10-4 and 6.4 × 10-2 for both adults and children. The finding suggests that consuming contaminated spring water could lead to severe acute, chronic, or sometimes fatal health consequences for the locals. This study provides valuable QMRA information on the contaminated karst groundwater, with important implications for human health and groundwater resources management.

High resolution parallel sequencing reveals multistrain Campylobacter in broiler chicken flocks testing ‘negative’ by conventional culture methods: implications for control of Campylobacter infection

Contaminated chicken meat is a major source of human Campylobacteriosis and rates of infection remain high, despite efforts to limit the colonisation of broiler (meat) chicken flocks on farms. Using conventional testing methods of culture or qPCR, Campylobacter is typically detected amongst broiler flocks from 3 wk of age, leading to the assumption that infection is introduced horizontally into chicken rearing houses at this time. In this study, we use parallel sequencing of a fragment of the Campylobacter outer membrane protein, encoded by the porA gene, to test for presence of Campylobacter DNA amongst fresh fecal samples collected from broiler flocks aged 23 to 28 d. Campylobacter DNA was detected in all of the 290 samples tested using the porA target, and in 48% of samples using 16S bacterial profiling, irrespective of whether or not Campylobacter could be detected using conventional qPCR thresholds. A single porAf2 variant was predominant among flocks that would be determined to be Campylobacter ‘positive’ by conventional means, but a diverse pattern was seen among flocks that were Campylobacter ‘negative’. The ability to routinely detect low levels of Campylobacter amongst broiler flocks at a much earlier age than would conventionally be identified requires a re-examination of how and when biosecurity measures are best applied for live birds. In addition, it may be useful to investigate why single Campylobacter variants proliferate in some broiler flocks and not others.

#67 Treatment of Campylobacter upsaliensis and Helicobacter canis in twin brothers with XLA

Abstract Background X-linked agammaglobulinemia (XLA) is a primary immunodeficiency associated with mutations in the B cell tyrosine kinase gene leading to failure in B cell maturation, resulting in antibody deficiency and increased susceptibility to infections. Patients with XLA are at risk of severe disease with infections with Campylobacter and closely related Helicobacter species and warrant treatment with close follow up given risk of recurrence. Method We present follow up of two 5-year-old fraternal twins with XLA who were diagnosed with C. upsaliensis and H. canis respectively via microbial cell-free DNA (mcfDNA) and underwent successful treatment with multi-drug antimicrobial therapy with 12 months of follow-up. Results Patient #1 was diagnosed with XLA at 11 months of age. At 3 years of age, he presented with fever and diarrhea, with stool cultures that grew C. jejuni. Despite treatment with azithromycin, his symptoms persisted and repeat stool cultures grew C. upsaliensis. Testing via mcfDNA confirmed C. upsaliensis. Patient #2 was diagnosed with XLA at 12 months of age. He developed fever, erythema nodosum (EN) like lesions and arthritis at 3.5 years old. Given his twin’s diagnosis of Campylobacter sp. enteritis, he was given a presumptive diagnosis of Campylobacter reactive arthritis and EN. Despite treatment with Naprosyn and 14 days of azithromycin, his symptoms persisted and stool PCR was positive for Campylobacter sp. mcfDNA testing confirmed C. upsaliensis and H.canis. Given failure of initial antimicrobial therapy, both boys were treated with a 4-week course of IV ertapenem, and oral ciprofloxacin and amoxicillin, and both developed neutropenia and concern for drug toxicity. Therapy was transitioned to triple drug therapy with oral doxycycline, ciprofloxacin and amoxicillin. Doxycycline was discontinued after 3 weeks due to patients’ young age. Ciprofloxacin and amoxicillin were continued for 12 months. Both boys had complete resolution of symptoms after 12 months of therapy and remain without recurrence 10 weeks after discontinuation of antimicrobial therapy. Conclusion Physicians should be aware of the treatment challenges of chronic Campylobacter and Helicobacter infection in boys with XLA and other immunocompromised hosts. Successful treatment may require prolonged multi-drug antimicrobial therapy, with monitoring for drug toxicities.

Campylobacter Gastroenteritis Among Under-Five Children in Southwest Ethiopia.

BackgroundUnder-five children are at an increased risk for foodborne illnesses because of the ingenuousness of their immune system. Although Campylobacter species are one of the bacterial etiologies of gastroenteritis, Campylobacter gastroenteritis among under-five children is not well considered in Ethiopia. Therefore, this study aimed at exploring the prevalence, associated risk factors, and antibiotic susceptibility patterns of Campylobacter species among under-five children with diarrhea.MethodsThe institution-based cross-sectional study was conducted among under-five children with diarrhea at Jimma Medical Center, southwestern Ethiopia from January 5 to April 21, 2020. Stool samples were collected and inoculated into Campylobacter agar medium. Isolation and identification were done using standard bacteriological techniques. Antibiotic susceptibility testing was conducted on Mueller-Hinton agar supplemented with 10% sheep blood using disk diffusion techniques. Bivariate and multivariate logistic regressions were used to assess the associated risk factors.ResultsA total of 214 under-five children were enrolled. The prevalence of Campylobacter infection was 8.9%. Absence of caretakers’ handwashing before preparation of food [AOR = 3.7, 95% CI: (1.2–10.8)], direct contact with domestic animals [AOR = 3.6, 95% CI: (1.0–12.7)], and consumption of raw dairy products [AOR = 4.5, 95% CI: (1.4–13.9)] are the factors associated with Campylobacter infection. Some Campylobacter species were found to be resistant to most available antibiotics.ConclusionThe magnitude of Campylobacter gastroenteritis indicates the need for routine isolation and identification of Campylobacter species from all under-five children clinically diagnosed with diarrhea. Species that are resistant to the drug of choice for Campylobacteriosis are also emerging. Health education on the importance of pasteurization of milk and caretakers’ handwashing can mitigate the transmission. Mechanism of handling of domestic animals should be considered to reduce transmission of zoonotic diseases like Campylobacteriosis.

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Validation of RapidChek®Campylobacter Test System for the Detection of C. jejuni, C. coli, and C. lari in Poultry Samples: AOAC Performance Tested MethodSM 052201.

Campylobacter is one of the leading causes of human bacterial gastroenteritis worldwide. Campylobacter infections are most often associated with the consumption of raw milk, undercooked poultry, and contaminated water. The RapidChek®Campylobacter test system (PTM number 052201) was validated for the detection of Campylobacter jejuni, C. coli, and C. lari in raw ground chicken, chicken carcass rinse, and turkey carcass sponges. The method uses a proprietary enrichment medium. Following aerobic enrichment, an immunochromatographic test strip is inserted into the tube containing the enrichment, developed for 20 min, and interpreted. Campylobacter-inoculated food samples were tested by the method, as well as the USDA/FSIS cultural reference method; Isolation and Identification of Campylobacter jejuni/coli/lari from Poultry Rinse, Sponge and Raw Product Samples MLG 41.04. The candidate method was also confirmed by an alternative cultural method. The RapidChek method was tested with 50 Campylobacter strains comprised of C. jejuni, C. coli, and C. lari, and 30 non-target strains. A total of 80 low-level spiked samples were tested by both methods in the study. The candidate method yielded 49 presumptive positives: all presumptive results were confirmed culturally. The reference method produced a total of 41 confirmed positive results. No difference between the alternate confirmation method and reference confirmation method was observed. Probability of detection analysis demonstrated no significant differences in the number of positive samples detected by the candidate method and cultural reference method. The RapidChek method detected all 50 Campylobacter strains and none of the 30 non-target strains, including Campylobacter spp. other than C. jejuni, C. coli, and C. lari. The candidate method performed as well as the reference method in the detection of C. jejuni, C. coli, and C. lari in raw ground chicken, chicken carcass rinse, and turkey carcass sponges. Aerobic enrichment of selected matrixes for 48 h yielded reliable presumptive results for Campylobacter.

P18 Antimicrobial resistance patterns of Campylobacter species isolated from poultry in Mathira, Nyeri County, Kenya

Background Campylobacter species are zoonotic pathogens recognized as a cause of gastroenteritis in the human population and pose a major public health threat worldwide. The most important in the genus is Campylobacter jejuni, which causes diarrhoea in patients, mainly children under the age of 1 year, young adults and immunosuppressed persons. Campylobacter occurs in the gut of most animals as normal flora and is transferred to foods from the faeces during slaughter. It is acquired through consumption of undercooked poultry and other meat as well as water and unpasteurized milk. Fruits and vegetables can be contaminated from water containing faeces from birds and other animals. Campylobacter can also be transmitted through contact with dogs and cats’ faeces as well as drinking contaminated water. Campylobacter infections are common in low- and middle-income countries as well as developed countries.ObjectivesTo assess the prevalence and determine the antimicrobial resistance patterns of Campylobacter spp. from poultry in Mathira Sub-County, Nyeri County.MethodsA total of 380 cloacal swabs were collected randomly form 57 farmers in Mathira, Kenya. They were transported in sterile Amies charcoal swabs under cold chain and stored at 4°C. They were enriched in Preston broth and incubated for 24 h at 42°C, after which they were streaked onto mCCDA agar and incubated for 24 h at 42°C under microaerobic conditions generated by Campy Gen™ packs. Typical grey moist, swarming and discrete colonies were identified using MALDI-TOF. Antimicrobial susceptibility testing was done on ampicillin, tetracycline, ciprofloxacin, gentamicin, erythromycin and nalidixic acid by the disc diffusion method and disc diameters were measured and interpreted using the CLSI guidelines.ResultsIn total, 272 out of 380 (71.32%) isolates were Campylobacter; 190 (50%) were C. jejuni while 81 (22) were Campylobacter coli. Resistance to ampicillin was 40% for C. coli and 30.53% for C. jejuni. Overall, 53.75% of C. coli isolates and 51.85% of C. jejuni isolates were resistant to tetracycline. We found 67.50% of C. coli were resistant to ciprofloxacin as compared with 38.42% of C. jejuni. Resistance to erythromycin for C. coli and C. jejuni was 10% and 16.935%, respectively. Finally, 65% of C. coli was resistant to nalidixic acid as compared with 38.42% of C. jejuni.ConclusionsThe study found that Campylobacter spp. show high levels of resistance to tetracyclines, ciprofloxacin and nalidixic acid and are MDR. Resistance to gentamicin and erythromycin is markedly low and these antibiotics can be reserved for treatment. Surveillance activities for AMR should be enhanced and laboratory capacity increased to support surveillance and diagnosis.

Prevalence and antimicrobial susceptibility pattern of Campylobacter jejuni in raw retail chicken meat in Metropolitan Accra, Ghana

Although, Campylobacter spp. are a major cause of foodborne gastroenteritis, its occurrence and antimicrobial resistance traits have not been well defined in low income countries, particularly in Africa. In this study, retail chicken was sampled (n = 400) between February 2019 to January 2020 in Metropolitan Accra, Ghana, to determine the prevalence and antimicrobial susceptibility pattern of Campylobacter jejuni. Raw chicken samples were obtained in wet markets (n = 315) and supermarkets (n = 85) and each subjected to direct plating and broth enrichment according to standard culture methods for Campylobacter spp. with the identity of presumptive positive colonies confirmed by MALDI-TOF. The susceptibility of isolates to antibiotics commonly used for campylobacteriosis in humans (in order to reflect the One Health significance of Campylobacter at the human-food interface) were then assessed by disc diffusion. A prevalence of 38.3% was recorded and all isolates were confirmed as Campylobacter jejuni. Enrichment yielded 127 positives while direct plating yielded 55 positives with low level of agreement in detection between these assays (Kappa = 0.15). Among samples positive by direct plating, the mean Campylobacter count was 1.9 log10 CFU/g (sd ±0.8). About 13% (7/55) of the samples positive by direct plating contained counts of 3log and above. Samples from the wet market yielded more positives than those from the supermarket with the rate of isolation from wet markets being 1.6 times that of the supermarket. Among 182 isolates characterized for their antimicrobial susceptibility, resistance to fluoroquinolones was 99.5%, tetracyclines 100% and macrolides 26.9%. Multi-drug resistance was also observed in 26.9% of the screened isolates. The findings point to a potential high level of exposure of humans to Campylobacter jejuni through chicken meat and thus the need for education on hygienic preparation and handling of raw chicken. High rates of resistance to classes of antimicrobials critically important for treating Campylobacter infections in humans; fluroquinolones and macrolides, affirm the need for stronger regulatory control of antimicrobials and enhanced antimicrobial stewardship in chicken production.

Antibiotic Resistance in Campylobacter spp. Isolated from Broiler Chicken Meat and Human Patients in Estonia.

Poultry meat is considered the most important source of Campylobacter spp. Because of rising antimicrobial resistance in Campylobacter spp., this study investigated the antimicrobial resistance of Campylobacter isolates from fresh broiler chicken meat originating from the Baltic countries sold in Estonian retail settings. Additionally, human clinical isolates obtained from patients with Campylobacter enteritis in Estonia were analysed. The aim of this study was to investigate the susceptibility of Campylobacter spp. to nalidixic acid, ciprofloxacin, tetracycline, streptomycin, erythromycin and gentamicin. The broth microdilution method with the EUCAMP2 panel was used for MIC determination and antimicrobial mechanisms were analysed using WGS data. A total of 46 Campylobacter strains were analysed, of which 26 (42.6%) originated from Lithuanian, 16 (26.2%) from Latvian, and 4 (6.6%) from Estonian fresh broiler chicken meat. In addition, 15 (24.6%) Campylobacter strains of patients with campylobacteriosis were tested. The antimicrobial resistance patterns of Campylobacter spp. isolated from fresh broiler chicken meat samples of Estonian, Latvian and Lithuanian origin collected in Estonian retail, and from patients with Campylobacter enteric infections, were determined. A total of 46 (75%) of the isolates tested were C. jejuni and 15 (25%) were C. coli. Campylobacter resistance was highest to nalidixic acid (90.2% of strains) and ciprofloxacin (90.2%), followed by tetracycline (57.4%), streptomycin (42.6%) and erythromycin (6.6%). All strains were sensitive to gentamicin. Additionally, antimicrobial resistance genes and point mutations were detected in 27 C. jejuni and 8 C. coli isolates previously assigned as resistant with the phenotypic method. A high antibiotic resistance of Campylobacter spp. in Lithuanian- and Latvian-origin broiler chicken meat and Estonian clinical isolates was found. Similar antibiotic resistance patterns were found for broiler chicken meat and human Campylobacter isolates.

Campylobacter jejuni from Slaughter Age Broiler Chickens: Genetic Characterization, Virulence, and Antimicrobial Resistance Genes

Campylobacter jejuni is a major cause of food-borne human gastroenteritis worldwide and is designated as a high priority antimicrobial-resistant pathogen by the World Health Organization (WHO). In this study, a total of 26 C. jejuni isolates from broiler chickens were screened for the presence of virulence and antimicrobial resistance genes by PCR. As a result, the study detected 11/26 (42.3%), 9/26 (34.6%), 8/26 (30.8%), 7/26 (26.9%), 6/26 (23.1%), and 6/26 (23.1%) of cdtC, pldA, cdtB, cdtA, cadF, and ciaB virulence genes, respectively, with seven of the isolates carrying more than two virulence genes. The majority of the isolates n = 25 (96.1%) were resistant to nalidixic acid, followed by n = 21 (80.7%), n = 22 (84.6%), and n = 5 (19.2%) for tetracycline, erythromycin, and ciprofloxacin, respectively. Most isolates were harboring catI (n = 16; 84.2%), catII (n = 15; 78.9%), catIII (n = 10; 52.6%), catIV (n = 2; 10.5%), floR (n = 10; 52.6%), ermB (n = 14; 73.7%), tetO (n = 13; 68.4%), tetA (n = 9; 47.4%), mcr-4 (n = 8; 42.1%), and ampC (n = 2; 10.5%). Meanwhile, mcr-1, mcr-2, mcr-3, mcr-5, tet(X), tet(P), and tet(W) genes were not detected in all isolates. Class I and Class II integrons were detected in 92.3% (n = 24) and 65.4% (n = 17) isolates, respectively. About 31% (8 of the 26 isolates) isolates were carrying more than two resistance genes. According to our knowledge, this is the first study to detect class II integrons in Campylobacter spp. (C. jejuni). The high prevalence of cdtA, cdtB, cdtC, cadF, pldA, and ciaB genes and antibiotic resistance genes in C. jejuni in this study indicates the pathogenic potential of these isolates. Majority of the isolates demonstrated resistance to nalidixic acid, tetracycline (tet), and erythromycin (ermB), which are the drugs of choice for treating Campylobacter infections. Therefore, these findings highlight the importance of implementing an efficient strategy to control Campylobacter in chickens and to reduce antimicrobial use in the poultry industry, which will help to prevent the spread of infections to humans.