Antimicrobial susceptibility in thermophilic Campylobacter species isolated from pigs and chickens in South Africa

Campylobacter jejuni is one of the leading causes of sporadic food-borne bacterial disease in humans. In intensive poultry and pig rearing systems the use of oral antibiotics is essential to maintain health. Consequently, there is a high risk for the thermophilic Campylobacter jejuni and C. coli resident in the intestinal tract of food animals to develop resistance to commonly used antibiotics. Contamination of meat or eggs with pathogenic strains of resistant Campylobacter could, therefore, result in a form of campylobacteriosis in humans that is difficult to treat. The aim of this investigation was to determine the antimicrobial susceptibility of thermophilic Campylobacter spp. isolated from pigs and poultry by the broth microdilution minimum inhibitory concentration (MIC) test. A total of 482 samples from the Western Cape and Gauteng provinces was collected and analysed. Thirty-eight Campylobacter isolates were obtained. Analysis of data revealed that C. jejuni strains mainly of poultry origin were more resistant to the fluoroquinolones, macrolides and tetracyclines and the C. coli strains were more resistant to the macrolides and lincosamides. Multi-resistance was also detected in 4 Campylobacter strains from the Western Cape. With the exception of tetracyclines, strains from high health Gauteng broiler farms were susceptible to antibiotics used to treat Campylobacter infections.     

Seasonal variation in Campylobacter-contaminated retail chicken products: a year-round investigation in Japan

Campylobacter was isolated from retail meat samples collected during the fiscal year 2009 in Japan. The higher percentages of contamination of chicken products were observed from June (39.3%) to November (83.3%). However, the highest number of human campylobacteriosis cases was reported in June in the Infectious Agents Surveillance Report. The chicken isolates with distinct clusters IVb and I, based on the restriction fragment length polymorphism of the flaA gene, were predominantly obtained during the periods between April and November 2009 and between February and March 2010, respectively. Extensive monitoring of Campylobacter contamination in chickens produced in various places is needed to analyse the seasonal variations between contamination of the meat products and the number of human cases with campylobacteriosis.     

Epidemiologic study of campylobacteriosis in Iowa cattle and the possible role of unpasteurized milk as a vehicle of infection

Bile samples were collected from 477 Iowa dairy cows and were cultured for thermophilic campylobacters. The prevalence of thermophilic campylobacters in the bile was 15.5%. Campylobacter jejuni and C coli from dairy cattle, chickens, pigs, sheep, and human beings were serotyped to develop host-species profiles. Human and cattle serologic profiles were the most similar, and human and chicken profiles shared several similarities. Epidemiologic data from 168 human cases of campylobacteriosis indicated that 23% of the cases were associated with consumption of unpasteurized milk.     

The prevalence and genetic diversity of Campylobacter spp. in domestic 'backyard' poultry in Canterbury, New Zealand

Campylobacteriosis is the most commonly notified illness in New Zealand. Whilst the importance of commercial poultry in campylobacteriosis is well established, little is known about the possible role of chickens kept at home as a direct animal/faecal contact or consumption exposure pathway. The aim of this study was to determine the prevalence and genetic diversity of Campylobacter spp. in domestic backyard chicken flocks in the Canterbury region of New Zealand. Poultry faecal samples were collected from 35 domestic 'backyard' poultry flocks from urban and rural properties around the Canterbury Region of New Zealand. A total of 291 samples were collected and tested for the presence of thermotolerant Campylobacter spp. and positive isolates were analysed using pulsed-field gel electrophoresis (PFGE) using both SmaI and KpnI enzymes. There was a high prevalence of Campylobacter spp. with 86% of flocks testing positive. Campylobacter jejuni alone, Campylobacter coli alone and both C. jejuni and C. coli were detected in 20 (57%), 2 (6%) and 8 (23%) of the flocks respectively. SmaI/KpnI PFGE analysis identified 50 different genotypes across the 35 flocks. Genotype diversity richness was highest on the lifestyle block and farm properties with 43 different genotypes isolated, whilst urban properties displayed the least richness with 12 genotypes isolated. Rural flocks tended to have more different genotypes in a given flock than urban flocks. Comparison of the genotypes with the PulseNet Aotearoa Campylobacter database showed that 28 of the genotypes had previously been isolated from human cases of campylobacteriosis. Many of these were also indistinguishable from Campylobacter spp. previously isolated from retail chicken. Therefore, contact with backyard poultry or their faecal material is a potential additional infection pathway outside of exposure to the established pathways associated with the consumption of Campylobacter-contaminated commercial meat or foods cross-contaminated from contaminated poultry.     

Prevalence of resistance to 11 antimicrobials among Campylobacter coill isolated from pigs on 80 grower-finisher farms in Ontario.

We carried out a cross-sectional study to investigate antimicrobial resistance patterns of Campylobacter coli isolated from Ontario grower-finisher pigs. From January to June 2004, 1200 samples were collected from 80 farms by obtaining a constant number (15) of fecal samples per farm. Susceptibility of the isolates to 11 antimicrobial drugs was determined by the agar-dilution technique. The overall prevalence of resistance to 1 or more antimicrobials among the isolates was 99.2%. High levels of resistance were observed for azithromycin, clindamycin, erythromycin, streptomycin, and tetracycline: 91.7%, 82.5%, 81.4%, 70.7%, and 63.7%, respectively. For sulfamethoxazole, ampicillin, and nalidixic acid, resistance was observed in 40.3%, 26.6%, and 22.7% of the isolates, respectively. Although at very low levels, resistance was observed for ciprofloxacin (a fluoroquinolone), chloramphenicol, and gentamicin: in 2.4%, 1.7%, and 0.2%, respectively. Many of the isolates (29.7%) were resistant to 5 antimicrobials, the most common being azithromycin, clindamycin, erythromycin, streptomycin, and tetracycline. Isolates from the same farm showed at least 5 patterns of resistance. Results from this study indicate high levels of resistance to the antimicrobial drugs most commonly used in the Canadian swine industry (macrolides, lincosamides, and tetracyclines) among C. coli isolated from grower-finisher pigs in Ontario. Macrolides and fluoroquinolones are the drugs most commonly used to treat severe human campylobacteriosis. Fortunately, at present, there is little resistance to fluoroquinolones among C. coli from pigs in Ontario.     

Fecal shedding of Campylobacter jejuni and Campylobacter coli among feral pigs in Texas

The population and range of feral pigs in the United States are rapidly expanding, yet key knowledge gaps exist regarding their role in the ecology and transmission of foodborne pathogens. Our objectives were to estimate the prevalence of Campylobacter jejuni and Campylobacter coli shedding among feral pigs throughout Texas and to identify risk factors for positive status. Faecal samples were collected from feral pigs in Texas from February 2014 through May 2015, and target organisms were detected using PCR assays. The prevalence of C. jejuni shedding was 1.6% (6/370), and the prevalence of C. coli shedding was 3.5% (13/370). C. coli shedding was significantly more common (p = .008) among female pigs than among male pigs. Feral pigs may represent a source of human campylobacteriosis.     

Genotypes and antibiotic resistance of canine Campylobacter jejuni isolates

Campylobacter jejuni is the most important cause of bacterial gastroenteritis in humans. It is a commensal in many wild and domestic animals, including dogs. Whereas genotypes of human and chicken C. jejuni isolates have been described in some detail, only little information on canine C. jejuni genotypes is available. To gain more information on genotypes of canine C. jejuni and their zoonotic potential, isolates from routine diagnostics of diarrheic dogs as well as isolates of a prevalence study in non-diarrheic dogs were analyzed. Prevalence of thermophilic Campylobacter among non-diarrheic dogs was 6.3% for C. jejuni, 5.9% for Campylobacter upsaliensis and 0.7% for Campylobacter coli. The C. jejuni isolates were genotyped by multi locus sequence typing (MLST) and flaB typing. Resistance to macrolides and quinolones was genetically determined in parallel. Within the 134 genotyped C. jejuni isolates 57 different sequence types (ST) were found. Five STs were previously unrecognized. The most common STs were ST-48 (11.2%), ST-45 (10.5%) and ST-21 (6.0%). Whereas no macrolide resistance was found, 28 isolates (20.9%) were resistant to quinolones. ST-45 was significantly more prevalent in diarrheic than in non-diarrheic dogs. Within the common time frame of isolation 94% of the canine isolates had a ST that was also found in human clinical isolates. In conclusion, prevalence of C. jejuni in Swiss dogs is low but there is a large genetic overlap between dog and human isolates. Given the close contact between human and dogs, the latter should not be ignored as a potential source of human campylobacteriosis.     

Prevalence of Campylobacter spp. and Yersinia spp. in the pig production

The aim of this study was to determine the prevalence of Campylobacter spp. and Yersinia spp. in a total of 1,040 faecal samples taken from animals at different ages from four farrowing and twelve fattening herds. In the farrowing unit, faeces were collected from 68 sows (faecal samples) and 256 suckling piglets (rectal swab samples). Further samples were collected from 362 growing and 354 finishing pigs (rectal swab samples). Additionally, 56 feed and environmental samples were collected. During the slaughtering process, 122 pigs and their carcasses respectively, were sampled three times. Finally, 86 meat and minced meat samples were taken from 34 retail stores. Campylobacter spp. were isolated in sows (33.8 %), piglets (80.9 %), growing (89.2 %) and finishing (64.7 %) pigs. Yersinia spp. were detected in growing (15.2 %) and finishing (13.3 %) pigs only. After twelve hours of chilling neither Campylobacter spp. nor Yersinia spp. were detected. In raw meat samples, Campylobacter spp. were isolated from one liver sample and Yersinia enterocolitica from two meat samples. Common slaughter techniques and hygiene procedures may be effective tools to reduce the risk of contamination and recontamination of meat products since Campylobacter spp. and Yersinia spp. were found only sporadically in raw meat samples.     

Prevalence and antimicrobial resistance of Campylobacter coli isolated from fattening pigs in France

Campylobacter are a leading cause of human diarrhea. The usual source of infection is contaminated food, particularly poultry but pork has also been described. The veterinary use of antimicrobial drugs has been suggested to be largely responsible for resistance in human isolates of this zoonotic pathogen. A study was carried out to investigate the occurrence and antimicrobial resistance of Campylobacter isolated from French fattening pigs. From March 1998 to June 1999, stomach samples were collected at slaughter from 240 fattening pigs originating from 24 different farms. Half of the pigs were found to be positive for Campylobacter but considerable variation was observed between farms. Isolates all belong to the Campylobacter coli species. Susceptibilities of the strains were determined for five antimicrobial drugs using agar dilution. Resistance to tetracycline and erythromycin was high (79 and 55%, respectively). For nalidixic acid, enrofloxacin and ampicillin, resistance was observed in 34, 15 and 20% of the isolates, respectively. More than one-third of the strains was resistant to at least three antimicrobial drugs. A Thr86Ile modification in GyrA was observed in the enrofloxacin-resistant strains studied. The multiresistant strains analyzed expressed the multidrug transporter CmeB at a high level. Results indicated a high prevalence of C. coli in the stomach of the French pigs examined. In addition, a high proportion of the strains was resistant to antimicrobial drugs, particularly to tetracycline and erythromycin, or were multiresistant.     

Poultry meat as a source of human campylobacteriosis

Campylobacteriosis is a prevalent diarrhoeal illness in humans, and poultry meat is considered the most important source of foodborne infection with Campylobacter spp. In this review, the disease burden of campylobacteriosis, the most important risk factors for Campylobacter colonization in broilers, and potential interventions to reduce poultry-related human exposure to Campylobacter spp. are discussed.     

Genotypic characterisation and cluster analysis of Campylobacter jejuni isolates from domestic pets, human clinical cases and retail food

The genetic similarity of Campylobacter jejuni isolates from pets, compared to human clinical cases and retail food isolates collected in Ireland over 2001-2006 was investigated by cluster analysis of pulsed-field gel electrophoresis (PFGE) fingerprinting profiles. Comparison of the PFGE profiles of 60 pet isolates and 109 human isolates revealed that seven (4.1%) profiles were grouped in clusters including at least one human and one pet C. jejuni isolate. In total six (1.6%) of 60 pet and 310 food profiles were in clusters with at least one food and one pet C. jejuni isolate. The detection of only a small number of genetically indistinguishable isolates by PFGE profile cluster analysis from pets and from humans with enteritis in this study suggests that pets are unlikely to be an important reservoir for human campylobacteriosis in Ireland. However, genetically indistinguishable isolates were detected and C. jejuni from pets may circulate and may contribute to clinical infections in humans. In addition, contaminated food fed to pets may be a potential source of Campylobacter infection in pets, which may subsequently pose a risk to humans.     

Occurrence, species distribution, antimicrobial resistance and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of domestic animals

beta-Lactams and macrolides are important antibiotics for treatment of staphylococcal infections in both humans and animals. The aim of the study was to investigate the occurrence, species distribution and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of dogs, horses, pigs, and cattle in Denmark. Nasal swabs were collected from a total of 400 animals, including 100 individuals of each species. Methicillin- and erythromycin-resistant staphylococci were isolated on selective media, identified by 16S rDNA sequencing, and typed by pulsed field gel electrophoresis (PFGE). Methicillin-resistant coagulase-negative staphylococci (MRCoNS) harbouring mecA were isolated from horses (50%) and dogs (13%), but not from food animals. The species identified were S. haemolyticus (n=21), S. vitulinus (n=19), S. sciuri (n=13), S. epidermidis (n=8), and S. warneri (n=2). mecA-mediated methicillin resistance in S. vitulinus was described for the first time. Methicillin-resistant S. aureus was not detected. PFGE analysis revealed the presence of specific MRCoNS clones in samples originating from the same veterinary hospital or equine farm. Erythromycin-resistant S. aureus (ERSA) was detected in 38% of pigs and all isolates harboured a constitutively expressed erm(C) gene. The vast majority (37/38) of pigs carrying ERSA originated from a farm characterized by frequent use of macrolides. Most ERSA isolates (28/38) displayed indistinguishable or closely related PFGE patterns, indicating clonal distribution within the farm. Based on the analysis of data on antimicrobial consumption, the occurrence of MRCoNS in companion animals and that of ERSA in pigs reflected national and local patterns of antimicrobial usage.     

Chronic pleuritis in Danish slaughter pig herds

Respiratory disease is considered the most serious disease problem in modern pig production and the risk has increased with intensification of pig production. We quantified risk factors for chronic pleuritis (CP) in Danish pig herds in terms of herd and herd-owner characteristics, management and neighbourhood factors. The occurrence of CP was investigated in 540,104 slaughter pigs from 259 farrow-to-finish or finishing herds during the mandatory post-mortem meat inspection at 18 Danish abattoirs. The monthly herd- and abattoir-specific prevalences of CP were estimated for the months January through August 2000. Meat-inspection data, herd characteristics and neighbourhood factors were obtained from databases at the Danish Bacon and Meat Council. Data on herd-owner characteristics and management factors were obtained by telephone interviews. Data were analysed using a mixed model accounting for repeated measurements. Four factors were associated with increased herd prevalence of CP: low health status of the herd, pig density within a 5 km radius, mingling of pigs during the production period and the month of slaughter. Two factors protected against CP: feeding with only dry feed and practising all-in-all-out production.     

Evaluation of the antibacterial residue surveillance programme in Danish pigs using Bayesian methods

Residues of pharmacological active substances or their metabolites might be found in food products from food-producing animals. Maximum Residue Limits for pharmacological active substances in foodstuffs of animal origin are established to assure high food safety standards. Each year, more than 20,000 samples are analysed for the presence of antibacterial residues in Danish pigs. This corresponds to 0.1% of the size of the slaughter pig population and more than 1% of the sows slaughtered. In this study, a Bayesian model was used to evaluate the Danish surveillance system accuracy and to investigate the impact of a potential risk-based sampling approach to the residue surveillance programme in Danish slaughter pigs. Danish surveillance data from 2005 to 2009 and limited knowledge about true prevalence and test sensitivity and specificity were included in the model. According to the model, the true antibacterial residue prevalence in Danish pigs is very low in both sows (～0.20%) and slaughter pigs (～0.01%). Despite data constraints, the results suggest that the current screening test used in Denmark presents high sensitivity (85-99%) and very high specificity (>99%) for the most relevant antibacterial classes used in Danish pigs. If high-risk slaughter pigs could be identified by taking into account antibacterial use or meat inspection risk factors, a potential risk-based sampling approach to antibacterial residue surveillance in slaughter pigs would allow reducing the sample size substantially, while increasing or maintaining the probability of detection. Hence, the antibacterial residue surveillance programme in Danish pigs would be more cost-effective than today.     

Prevalence and shedding patterns of Campylobacter spp. in longitudinal studies of kennelled dogs

Dog ownership is considered a risk factor for campylobacteriosis in humans. This study investigated the prevalence and shedding of Campylobacter spp. in kennelled dogs. Faecal samples (n=399) were collected in longitudinal studies from 52 dogs in two kennels. Campylobacter spp. were isolated using charcoal-based selective agars and direct PCR. The prevalence of Campylobacter spp. in dogs in boarding kennels ranged from 46% (95% CI 22, 72) on entry, to 50% (95% CI 30, 70) overall, and in dogs in 'rescue' kennels from 68% (95% CI 49, 84) on entry to 73% (95%, CI 56, 87) overall. C. upsaliensis was isolated from 62% (95% CI 48, 73) of the dogs, whilst C. jejuni was isolated from 15% (95% CI 7, 26) of animals. The majority of infected dogs entered the kennels already carrying Campylobacter spp., and remained infected throughout their stay. However, in some cases, shedding appeared to commence after kennelling. Given that the prevalence of C. upsaliensis and C. jejuni was relatively high in dogs from both boarding and rescue kennels, such animals may pose a zoonotic risk.     

Prevalence of Campylobacter jejuni in selected domestic and wild birds in Louisiana

Prevalence of Campylobacter jejuni was determined in a selected population of domestic and free-living birds submitted for necropsy to the Louisiana State Veterinary Medical Diagnostic Laboratory. The 445 cases examined included 13 orders of birds and yielded C. jejuni in 45 cases, representing an isolation rate of 10.1%. Prevalence was highest in Galliformes (25.2%), followed by Anseriformes (12.9%) and Columbiformes (8.3%). Only one isolation was made out of 179 Psittaciformes examined. Penner serotypes 1, 2, 4, and 16 were most commonly identified among the C. jejuni isolates. This study emphasizes the importance of Galliformes as reservoirs of C. jejuni. The commonality of these serotypes with isolates derived from humans suggests the zoonotic potential of Galliformes in relation to human campylobacteriosis. The isolation rate of 12.9% in Anseriformes implicates free-living and migratory waterfowl as carriers of C. jejuni. Results confirm that Psittaciformes represent a low risk of C. jejuni infection in humans.     

Utilizing a combination of molecular and spatial tools to assess the effect of a public health intervention

Until recently New Zealand had one of the highest rates of human campylobacteriosis reported by industrialized countries. Since the introduction of a range of control measures in the poultry production chain a reduction in human cases of around 50% has been observed nationwide. To inform risk managers a combination of spatial, temporal and molecular tools - including minimum spanning trees, risk surfaces, rarefaction analysis and dynamic source attribution modelling - was used in this study to formally evaluate the reduction in disease risk that occurred after the implementation of control measures in the poultry industry. Utilizing data from a sentinel surveillance site in the Manawatu region of New Zealand, our analyses demonstrated a reduction in disease risk attributable to a reduction in the number of poultry-associated campylobacteriosis cases. Before the implementation of interventions poultry-associated cases were more prevalent in urban than rural areas, whereas for ruminant-associated cases the reverse was evident. In addition to the overall reduction in prevalence, this study also showed a stronger intervention effect in urban areas where poultry sources were more dominant. Overall a combination of molecular and spatial tools has provided evidence that the interventions aimed at reducing Campylobacter contamination of poultry were successful in reducing poultry-associated disease and this will inform the development of future control strategies.     

Methods for investigating Trichinella infections in domestic and wild animals

Trichinellosis is an important parasitic zoonosis that is caused by the intracellular nematode Trichinella spp.. Infection of humans occurs through consumption of raw (or undercooked) meat containing infectious larvae. In Europe, meat from pork, horse, and wild boar have been identified as most important sources of Trichinella infections in humans. In Switzerland, both the domestic pig and wild boar population are considered free of Trichinella. Conversely, Swiss foxes, lynxs and recently a wolf were found to be infected, the species identified in these animals was always referred to as Trichinella britovi. Although this species rarely infects pork and, compared to Trichinella spiralis, only causes reduced pathogenic effects in humans, the basic presence of Trichinella in Switzerland cannot be neglegted. This fact has gained increasing importance since the responsible authorities in the European Union (EU) are preparing regulations for the official Trichinella-control in meat in order to improve food safety for consumers. These regulations will be implemented as a consequence of the recent association of east European countries with the EU. This new legislation particularly takes into account, that in the past by far most cases of human trichinellosis in the EU were due to consumption of imported east European meat.Within the framework of the bilateral agreements of Switzerland with the EU, the Swiss veterinary public health authorities will have to comply with the foreseen EU regulations. Although diagnostic methods for the direct demonstation of Trichinella in pork meat are already routine practice in several Swiss abattoirs, the implementation of a meat control program for Trichinella for the entire slaughter pig population of the country would lead to an enormous increase in costs for the administration and will require an increased infrastructure in veterinary services. In order to find a reduced testing format for monitoring Trichinella infections in Swiss pork, an infection risk-oriented survey strategy is currently evaluated. In the present article, this minimized survey strategy is discussed regarding its compatibility with the EU regulations laying down rules for the official control of meat for Trichinella.     

Antimicrobial susceptibility patterns of thermotolerant Campylobacter strains isolated from food animals in Ethiopia

Thermotolerant Campylobacter spp. are frequent causes of diarrhoea in humans worldwide mostly originating from poultry. It has been suggested that extensive veterinary use of antibiotics is largely responsible for resistance in human isolates. During a 4-month period from January to April 2004, 192 Campylobacter spp. were isolated from fecal samples of 485 healthy food animals. The in vitro susceptibility to 12 antibiotics was determined by the agar disk diffusion method. Among the 192 Campylobacter spp. isolated, 135 (70.3%) were identified to be C. jejuni, 51 (26.6%) were C. coli and 6 (3.1%) were C. lari. C. jejuni was the most prevalent species in chickens (80.8%) versus 16.2% C. coli and 3.0% C. lari. All isolates found in pigs were C. coli. All strains were sensitive to chloramphenicol and ciprofloxacin and all were resistant to cephalothin. More than 90% of the strains were sensitive to clindamycin, erythromycin, gentamicin, nalidixic acid, norfloxacin, streptomycin and tetracycline. Resistance was found against ampicillin in 20% and trimethoprim-sulphamethoxazole in 37.5%. Resistance was not statistically different among C. jejuni, C. coli and C. lari (p>0.05). Multidrug resistance to two or more drugs was detected in 14.5% of strains. In conclusion, the study showed that antimicrobial resistance is found only at relatively low frequencies for most antimicrobial agents tested except for ampicillin and trimethoprim-sulphamethoxazole. The low percentages of resistance to most antimicrobial agents tested in this study may be the result of low/no usage of these agents as a growth promoters or treatment in the Ethiopian animal farm setting. The detection of multidrug resistant isolates may pose a threat to humans and further limits therapeutic options.     

PFGE and PCR/RFLP typing of Campylobacter jejuni strains from poultry

1. Pulsed-field gel electrophoresis (PFGE) and PCR-restriction fragment length polymorphisms of the flagellin gene (fla-RFLP) were used to analyse 92 poultry and 110 human strains of Campylobacter jejuni. 2. Among poultry strains, 11 fla-RFLP and 11 PFGE subtypes were found, while human strains could be divided into 23 fla-RFLP and 32 PFGE subtypes. Altogether, 31 fla-RFLP and 32 PFGE subtypes were found. 3. The results show that individual flocks in farms are mostly infected with a single C. jejuni clone, while during subsequent colonisation their genotypes altered. fla-RFLP and PFGE profiles in poultry and humans were identical in less than 6% of cases. The results found so far confirm previous findings that chicken meat does not represent as important a source of campylobacteriosis as was previously believed. 4. The typing of Campylobacter sp. forms the basis for an evaluation of the current state and risk assessment of various Campylobacter sp. sources in relation to humans. Examination of samples with only one method is insufficient for epidemiology studies, because apparently different clones identified with one method could originate from a single clone, which could be proved with the other method.