ABSTRACT. Campylobacter jejuni is the major cause of human gastroenteritis worldwide. Under stress conditions, C. jejuni can enter a viable but non-culturable (VBNC) state. We found that the C. jejuni was able to enter a VBNC state by prolonged incubation at 4C. The standard isolation methods using pre-enrichment steps in Bolton broth or Preston broth could not detect the VBNC cells in spiked chicken meat. The transcription levels of virulence-associated genes (flaA, flaB, cadF, ciaB, cdtA, cdtB and cdtC) were expressed in VBNC cells but in low levels. The VBNC cells retained the ability to invade Caco-2 human intestinal epithelial cells in vitro. In most cases, VBNC cells failed to resuscitate in Caco-2 cells, but in some experiments, they formed colonies after co-incubation with host cells. Collectively, C. jejuni enters into a VBNC state at 4C and the VBNC C. jejuni remains virulent which may possibly lead to disease in humans. C. jejuni in VBNC state is a potential concern for food safety.KEY WORDS: Campylobacter jejuni, resuscitation, viable but nonculturable state, virulence.

The prevalence of Campylobacter jejuni in wild birds is a potential hazard for human and animal health. The aim of this study was to establish the prevalence of C. jejuni in wild birds in Tokachi area, Hokkaido, Japan and investigate their virulence in vitro. In total, 173 cloacal swabs from individual wild birds were collected for the detection of Campylobacter spp. Thirty four samples (19.7%) were positive for Campylobacter of which 94.1% (32/34 samples) were C. jejuni. Additionally, one C. coli and one C. fetus were isolated. Seven C. jejuni isolates (one from crows and the other from pigeons) had important virulence genes including all three CDT genes (cdtA, cdtB and cdtC) and flaA, flaB, ciaB and cadF, and the other isolates were lacking cdtA gene. Further studies on in vitro virulence-associated phenotypes, such as motility assay on soft agar and invasion assay in Caco-2 cells, were performed. The wild bird C. jejuni isolates adhered and invaded human cells. Although the numbers of viable intracellular bacteria of wild bird isolates were lower than a type strain NCTC11168, they persisted at 48-hr and underwent replication in host cells.

BackgroundAiming for early disease detection and prompt outbreak control, digital technology with a participatory One Health approach was used to create a novel disease surveillance system called Participatory One Health Disease Detection (PODD). PODD is a community-owned surveillance system that collects data from volunteer reporters; identifies disease outbreak automatically; and notifies the local governments (LGs), surrounding villages, and relevant authorities. This system provides a direct and immediate benefit to the communities by empowering them to protect themselves.ObjectiveThe objective of this study was to determine the effectiveness of the PODD system for the rapid detection and control of disease outbreaks.MethodsThe system was piloted in 74 LGs in Chiang Mai, Thailand, with the participation of 296 volunteer reporters. The volunteers and LGs were key participants in the piloting of the PODD system. Volunteers monitored animal and human diseases, as well as environmental problems, in their communities and reported these events via the PODD mobile phone app. LGs were responsible for outbreak control and provided support to the volunteers. Outcome mapping was used to evaluate the performance of the LGs and volunteers.ResultsLGs were categorized into one of the 3 groups based on performance: A (good), B (fair), and C (poor), with the majority (46%,34/74) categorized into group B. Volunteers were similarly categorized into 4 performance groups (A-D), again with group A showing the best performance, with the majority categorized into groups B and C. After 16 months of implementation, 1029 abnormal events had been reported and confirmed to be true reports. The majority of abnormal reports were sick or dead animals (404/1029, 39.26%), followed by zoonoses and other human diseases (129/1029, 12.54%). Many potentially devastating animal disease outbreaks were detected and successfully controlled, including 26 chicken high mortality outbreaks, 4 cattle disease outbreaks, 3 pig disease outbreaks, and 3 fish disease outbreaks. In all cases, the communities and animal authorities cooperated to apply community contingency plans to control these outbreaks, and community volunteers continued to monitor the abnormal events for 3 weeks after each outbreak was controlled.ConclusionsBy design, PODD initially targeted only animal diseases that potentially could emerge into human pandemics (eg, avian influenza) and then, in response to community needs, expanded to cover human health and environmental health issues.

This study was conducted to determine the prevalence and quantify the number of Listeria monocytogenes in fresh chicken carcasses sold in traditional markets and supermarkets in Bandung, West Java, Indonesia, and to determine the antimicrobial resistance patterns of the isolated L. monocytogenes strains. The overall prevalence of L. monocytogenes in chicken carcasses was 15.8% (29/184). When comparing samples from traditional markets and supermarkets, no significant difference in the L. monocytogenes prevalence was detectable (15.2 versus 16.3%). Of the samples, 97.3% had L. monocytogenes counts <100 CFU/g, 2.2% had L. monocytogenes counts between 101 and 1,000 CFU/g, and 0.5% had L. monocytogenes counts of 1,001 to 10,000 CFU/g. Of the isolates, 27.6% were resistant to at least one of the 10 antimicrobials tested, with the major resistant phenotypes to penicillin (17.2%), ampicillin (6.9%), and erythromycin (6.9%). All 29 isolates recovered in this study were grouped into the molecular serogroup IIb, comprising the serovars 1/2b, 3b, and 7.

Many developing countries have recently faced public health challenges due to outbreaks of human rabies, while some areas have remained free from outbreaks of the disease. The aim of this study was to identify keys to good knowledge, attitude and practice (KAP) in rabies-free areas of Thailand to provide a functional model for rabies prevention in endemic zones. A cross-sectional study was conducted using a questionnaire in conjunction with interviews that were designed to elicit relevant information. Multivariate logistic regression analysis and content analysis were used to interpret the data obtained from the questionnaire and interviews, respectively. The results gathered from 1,392 respondents and 36 interviewees indicated that age, education and the residential area of the subjects were associated with knowledge and practice, but not with attitude. Late midlife subjects reported good knowledge and practice scores due to some form of personal encounter with rabies and attendance at rabies meetings and campaigns, while secondary education levels or above were associated with higher knowledge and practice scores due to the inclusion of a rabies course in the secondary school curriculum. Moreover, the findings showed that a community using a form of public communication known as wired broadcasting had greater knowledge, and a community organizing comprehensive participatory activities had better practice. Ultimately, it was not only age, education and residential area, but also the integration of appropriate interventions and control measures by the authorities that proved beneficial in making good knowledge and practice more uniform throughout the communities included in the study.

BackgroundRabies is a vaccine-preventable neglected tropical viral zoonosis. It occurs worldwide, creating a very heavy burden in many developing countries, including Nepal. Dogs are the principle vector for the transmission of this disease in urban areas. Vaccination is the most important preventive measure in areas where dogs are the principle source of infection. This study was conducted with the aim of detecting virus-neutralising antibodies and associated factors against rabies in vaccinated household dogs of Kathmandu valley. MethodsBlood samples were collected from 110 vaccinated pet dogs in Kathmandu, Bhaktapur, and Lalitpur districts of Nepal. The samples were taken to the laboratory of the National Zoonosis and Food Hygiene Research Center where serum was separated. An indirect immuneenzymatic assay (Platelia TM Rabies II kit ad usum Veterinarium, Biorad, China) was used for the detection of rabies virus anti-glycoprotein antibodies in the dog serum samples following the manufacturer's recommendations and instructions. Optical density values for unknown samples were compared with the positive sera titers in quantification tests obtained after a direct reading on the standard curve. Results were expressed as equivalent units per ml (EU/ml). FindingsOf the total samples, 89.09% exceeded the required seroconversion level (� 0.5 EU/ml); another 9.09% did not reach the seroconversion level (0.125-0.5 EU/ml); and 1.81% had undetectable seroconversion levels (<0.125 EU/ml) suggesting that the animal had not PLOS ONE

This study was performed to determine the prevalence and to semiquantify Campylobacter spp. on chicken meat samples at 4 selected local wet markets in Nueva Ecija, Philippines, and to determine the antimicrobial resistance patterns of the Campylobacter isolates. Out of 120 chicken meat samples, 57 (47.5%) were Campylobacter spp. positive. The majority of isolated Campylobacter strains were identified as Campylobacter coli (54.4%) and 45.6% as Campylobacter jejuni. Most of these positive samples (52.6%) showed a very high quantitative Campylobacter contamination (most probable number > 2,400/g, lower confidence limit 580/g). For antimicrobial resistance testing, 44 C. coli/jejuni isolates were tested using the agar disk diffusion method. Out of these, 77.3% were resistant to ampicillin, followed by ciprofloxacin (70.4%), tetracycline (54.6%), erythromycin (20.2%), and gentamicin (11.4%). Of the isolates, 36.4% (n = 16) were resistant to 1 antimicrobial agent, 34.1% (n = 15) were resistance to 3 antimicrobial agents, 13.6% (n = 6) to 2 antimicrobial agents, 9.1% (n = 4) to 4 antimicrobial agents, and 6.8% (n = 3) to all 5 antimicrobial agents tested. Our data demonstrate a high contamination of fresh chicken meat with Campylobacter spp. at retail in the Philippines. The detected high Campylobacter prevalences and quantitative loads on chicken meat at retail in the Philippines highlight the need to implement efficient intervention measures along the food chain and to encourage sanitary handling of poultry meat.

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