Dr Juan Carrique-Mas

Research Area: Global Health
Technology Exchange: Medical statistics
Scientific Themes: Clinical Trials & Epidemiology and Tropical Medicine & Global Health
Keywords: Zoonoses, Antimicrobial resistance, Veterinary epidemiology, Food-borne infections and VIetnam
Web Links:

Unfortunately, many antimicrobial products have lost their efficacy to treat human and animal disease, since bacteria have become unresponsive to these products. This phenomenon (termed ‘antimicrobial resistance’, or AMR) is a concern worldwide, and generally results from excessive and inappropriate usage of antimicrobials. In animal production, this results in the reduction of our weaponry to effectively treat animal bacterial diseases, leading to severe economic losses on farms. Many of these resistant bacteria or genes encoding for resistant on farms may also be transmitted to humans, resulting in untreatable human disease. ViParc aims at helping farmers in the Mekong Delta of Vietnam raise meat chickens using lesser amounts of antimicrobials.

ViParc study design

ViParc stands for ‘Vietnamese Platform for Antimicrobial Reductions in Chicken production’. ‘Vi’ also stands for ‘Veterinary intervention’. The ViParc project is funded by the Wellcome Trust (Grant Reference No.: 110085/Z/15/Z).

ViParc addresses this problem through a farm-based scientific ‘trial’, where farms from the area are randomly selected, and observed over a period of time, including a ‘baseline phase’ followed by an ‘intervention phase’. The study is to be conducted over three years in the Mekong Delta province of Dong Thap (Vietnam). Results will be fully evaluated at the end of the intervention phase.

The proposed intervention (delivered through the ‘intervention phase’) will consist of the delivery of a local veterinary support system to those farmers randomly assigned to the ‘intervention arm’, to help them improve farm productivity and reduce disease, therefore reducing also their reliance on antimicrobials. Farmers in the intervention group will receive free training on good husbandry practices, including the prevention and control of poultry diseases, as well as local veterinary support free of charge. In addition, half of the farmers in the intervention group will be asked not to use antimicrobials in feed. We will provide to these farmers with suitable alternatives to prevent disease that may appear as a result of the feed change. Because this study is designed as a scientific trial, a ‘control arm’ is required so that the effect of the intervention can be appropriately measured. This control arm will consist of farms that will not receive free veterinary support. The design of the ViParc study is summarised in the following chart:

Throughout the ViParc project we will collect chicken faecal samples at different stages of production using paper liners. These samples will be investigated in the laboratory for antimicrobial resistance in ‘commensal’ bacteria (i.e. bacteria that do not cause disease problems in chickens) of the gastrointestinal tract of the chickens (Escherichia coli). We will also investigate antimicrobial residues in chicken meat at the end of the production cycle of each crop, and we will take environmental samples as well as post cleaning and disinfection samples in some farms. We will also conduct data collection to help us understand: (1) antimicrobial usage; (2) disease and productivity; (3) levels of antimicrobial resistance in E. coli; and (4) residues of antimicrobials in the meat of the chickens raised.

Research questions

ViParc will try to address, among others, the following:

1. How and when do farmers use antimicrobials (including in feed) and what is their knowledge about these medicines?

2. How do chicken bacteria become resistant against antimicrobials? What are the most common types of resistance in chicken farms?

3. What is the contribution of hatcheries and inadequate cleaning and disinfection on antimicrobial resistance?

4. Does meat from chickens raised in the study farms contain antimicrobial residues?

5. Does veterinary advice help farmers reduce disease in chicken flocks?

A multidisciplinary team

ViParc is a multidisciplinary research project aimed at addressing antimicrobial resistance in animal production. This is highly pressing ‘One Health’ issue not only in Vietnam, but worldwide. The project, funded by the Wellcome Trust Major Overseas Programme (United Kingdom), is led by Oxford University Clinical Research Unit (OUCRU) in Ho Chi Minh City and implemented in Dong Thap province through the Sub-Department of Animal Health and Production (SDAHP-DT). The project includes specialists in Veterinary Medicine, Microbiology and Laboratory Science, Animal Health Economics and Public Engagement. Collaborators of ViParc include the University of Liverpool (United Kingdom) (Prof. Jonathan Rushton), Chulalongkorn University (Thailand) (Assoc. Prof. Niwat Chansiripornchai), the University of Barcelona (Spain) (Dr. Alexis Ribas), and the University of Can Tho (Prof. Viet Thu Ho Thi). The project has an important Public Engagement component, supported by the Wellcome Trust. The study has been approved by the People’s Committed in Dong Thap, by the review board of SDAH-DT, as well as by the OXTREC (Ethics Committee of the University of Oxford) (No. 5121-16).

Name Department Institution Country
Professor Jonathan Rushton Royal Veterinary College United Kingdom
Professor Ho Thi Viet Thu Microbiology University of Can Tho Vietnam
Alexis Ribas University of Barcelona Spain
Niwat Chansiripornchai Chulalongkorn University Thailand
Zellweger RM, Carrique-Mas J, Limmathurotsakul D, Day NPJ, Thwaites GE, Baker S, Southeast Asia Antimicrobial Resistance Network. 2017. A current perspective on antimicrobial resistance in Southeast Asia. J Antimicrob Chemother, 72 (11), pp. 2963-2972. | Show Abstract | Read more

Southeast Asia, a vibrant region that has recently undergone unprecedented economic development, is regarded as a global hotspot for the emergence and spread of antimicrobial resistance (AMR). Understanding AMR in Southeast Asia is crucial for assessing how to control AMR on an international scale. Here we (i) describe the current AMR situation in Southeast Asia, (ii) explore the mechanisms that make Southeast Asia a focal region for the emergence of AMR, and (iii) propose ways in which Southeast Asia could contribute to a global solution.

Nhung NT, Chansiripornchai N, Carrique-Mas JJ. 2017. Antimicrobial Resistance in Bacterial Poultry Pathogens: A Review. Front Vet Sci, 4 (AUG), pp. 126. | Show Abstract | Read more

Antimicrobial resistance (AMR) is a global health threat, and antimicrobial usage and AMR in animal production is one of its contributing sources. Poultry is one of the most widespread types of meat consumed worldwide. Poultry flocks are often raised under intensive conditions using large amounts of antimicrobials to prevent and to treat disease, as well as for growth promotion. Antimicrobial resistant poultry pathogens may result in treatment failure, leading to economic losses, but also be a source of resistant bacteria/genes (including zoonotic bacteria) that may represent a risk to human health. Here we reviewed data on AMR in 12 poultry pathogens, including avian pathogenic Escherichia coli (APEC), Salmonella Pullorum/Gallinarum, Pasteurella multocida, Avibacterium paragallinarum, Gallibacterium anatis, Ornitobacterium rhinotracheale (ORT), Bordetella avium, Clostridium perfringens, Mycoplasma spp., Erysipelothrix rhusiopathiae, and Riemerella anatipestifer. A number of studies have demonstrated increases in resistance over time for S. Pullorum/Gallinarum, M. gallisepticum, and G. anatis. Among Enterobacteriaceae, APEC isolates displayed considerably higher levels of AMR compared with S. Pullorum/Gallinarum, with prevalence of resistance over >80% for ampicillin, amoxicillin, tetracycline across studies. Among the Gram-negative, non-Enterobacteriaceae pathogens, ORT had the highest levels of phenotypic resistance with median levels of AMR against co-trimoxazole, enrofloxacin, gentamicin, amoxicillin, and ceftiofur all exceeding 50%. In contrast, levels of resistance among P. multocida isolates were less than 20% for all antimicrobials. The study highlights considerable disparities in methodologies, as well as in criteria for phenotypic antimicrobial susceptibility testing and result interpretation. It is necessary to increase efforts to harmonize testing practices, and to promote free access to data on AMR in order to improve treatment guidelines as well as to monitor the evolution of AMR in poultry bacterial pathogens.

Hoa LNM, Tuan NA, My PH, Huong TTK, Chi NTY, Hau Thu TT, Carrique-Mas J, Duong MT, Tho ND, Hoang ND et al. 2017. Assessing evidence for avian-to-human transmission of influenza A/H9N2 virus in rural farming communities in northern Vietnam. J Gen Virol, 98 (8), pp. 2011-2016. | Show Abstract | Read more

Rural farming communities in northern Vietnam do not routinely practice vaccination for influenza A viruses (IAV) for either humans or poultry, which enables us to study transmission intensity via seroepidemiology. Using samples from a longitudinal cohort of farming households, we determined the number of symptomatic and asymptomatic human infections for seasonal IAV and avian A/H9 over 2 years. As expected, we detected virologically confirmed acute cases of seasonal IAV in humans, as well as large numbers of subclinical seroconversions to A/H1pdm [55/265 (21 %)], A/H3 [95/265 (36 %)] and A/H9 [24/265 (9 %)]. Five of the A/H9 human seroconverters likely represented true infections rather than heterosubtypic immunity, because the individuals seroconverted solely to A/H9. Among co-located poultry, we found significantly higher seroprevalance for A/H5 compared to A/H9 in both chickens and ducks [for northern study sites overall, 337/1105 (30.5 %) seropositive for A/H5 and 123/1105 (11.1 %) seropositive for A/H9].

Carrique-Mas JJ, Rushton J. 2017. Integrated Interventions to Tackle Antimicrobial Usage in Animal Production Systems: The ViParc Project in Vietnam Frontiers in Microbiology, 8 (JUN), | Show Abstract | Read more

© 2017 Carrique-Mas and Rushton. Antimicrobial usage and antimicrobial resistance (AMR) in animal production is now recognized to be an important contributor to the global problem of AMR. Initiatives to curb indiscriminate antimicrobial use in animal production are currently being discussed in many low- and middle-income countries. Well-designed, scientifically sound interventions aimed to tackle excessive antimicrobial usage should provide scientists and policy makers with evidence of the highest quality to guide changes in policy and to formulate better targeted research initiatives. However, since large-scale interventions are costly, they require careful planning in order not to waste valuable resources. Here, we describe the components of the ViParc project (www.viparc.org), one of the first large-scale interventions of its kind to tackle excessive antimicrobial usage in Southeast Asian animal production systems. The project has been formulated as a "randomized before-and-after controlled study" targeting small-scale poultry farms in the Mekong Delta region of Vietnam. It aims to provide farmers with a locally-adapted veterinary support service to help them reduce their reliance on antimicrobials. ViParc has been developed in the backdrop of efforts by the Government of Vietnam to develop a National Action Plan to reduce Antimicrobials in Livestock and Aquaculture. Crucially, the project integrates socio-economic analyses that will provide insights into the drivers of antimicrobial usage, as well as an assessment of the cost-effectiveness of the proposed intervention. Information generated from ViParc should help the Government of Vietnam refine its policies to curb excessive antimicrobial usage in poultry production, while lessons from ViParc will help tackle excessive antimicrobial usage in other productions systems in Vietnam and in the broader Southeast Asian region.

Berto A, Pham HA, Thao TTN, Vy NHT, Caddy SL, Hiraide R, Tue NT, Goodfellow I, Carrique-Mas JJ, Thwaites GE et al. 2017. Hepatitis E in southern Vietnam: Seroepidemiology in humans and molecular epidemiology in pigs. Zoonoses Public Health, | Show Abstract | Read more

Viral pathogens account for a significant proportion of the burden of emerging infectious diseases in humans. The Wellcome Trust-Vietnamese Initiative on Zoonotic Infections (WT-VIZIONS) is aiming to understand the circulation of viral zoonotic pathogens in animals that pose a potential risk to human health. Evidence suggests that human exposure and infections with hepatitis E virus (HEV) genotypes (GT) 3 and 4 results from zoonotic transmission. Hypothesising that HEV GT3 and GT4 are circulating in the Vietnamese pig population and can be transmitted to humans, we aimed to estimate the seroprevalence of HEV exposure in a population of farmers and the general population. We additionally performed sequence analysis of HEV in pig populations in the same region to address knowledge gaps regarding HEV circulation and to evaluate if pigs were a potential source of HEV exposure. We found a high prevalence of HEV GT3 viral RNA in pigs (19.1% in faecal samples and 8.2% in rectal swabs) and a high HEV seroprevalence in pig farmers (16.0%) and a hospital-attending population (31.7%) in southern Vietnam. The hospital population was recruited as a general-population proxy even though this particular population subgroup may introduce bias. The detection of HEV RNA in pigs indicates that HEV may be a zoonotic disease risk in this location, although a larger sample size is required to infer an association between HEV positivity in pigs and seroprevalence in humans.

Berto A, Anh PH, Carrique-Mas JJ, Simmonds P, Van Cuong N, Tue NT, Van Dung N, Woolhouse ME, Smith I, Marsh GA et al. 2017. Detection of potentially novel paramyxovirus and coronavirus viral RNA in bats and rats in the Mekong Delta region of southern Viet Nam. Zoonoses Public Health, | Show Abstract | Read more

Bats and rodents are being increasingly recognized as reservoirs of emerging zoonotic viruses. Various studies have investigated bat viruses in tropical regions, but to date there are no data regarding viruses with zoonotic potential that circulate in bat and rat populations in Viet Nam. To address this paucity of data, we sampled three bat farms and three wet markets trading in rat meat in the Mekong Delta region of southern Viet Nam. Faecal and urine samples were screened for the presence of RNA from paramyxoviruses, coronaviruses and filoviruses. Paramyxovirus RNA was detected in 4 of 248 (1%) and 11 of 222 (4.9%) bat faecal and urine samples, respectively. Coronavirus RNA was detected in 55 of 248 (22%) of bat faecal samples; filovirus RNA was not detected in any of the bat samples. Further, coronavirus RNA was detected in 12 of 270 (4.4%) of rat faecal samples; all samples tested negative for paramyxovirus. Phylogenetic analysis revealed that the bat paramyxoviruses and bat and rat coronaviruses were related to viruses circulating in bat and rodent populations globally, but showed no cross-species mixing of viruses between bat and rat populations within Viet Nam. Our study shows that potentially novel variants of paramyxoviruses and coronaviruses commonly circulate in bat and rat populations in Viet Nam. Further characterization of the viruses and additional human and animal surveillance is required to evaluate the likelihood of viral spillover and to assess whether these viruses pose a risk to human health.

Trung NV, Matamoros S, Carrique-Mas JJ, Nghia NH, Nhung NT, Chieu TTB, Mai HH, van Rooijen W, Campbell J, Wagenaar JA et al. 2017. Zoonotic Transmission of mcr-1 Colistin Resistance Gene from Small-Scale Poultry Farms, Vietnam. Emerg Infect Dis, 23 (3), pp. 529-532. | Show Abstract | Read more

We investigated the consequences of colistin use in backyard chicken farms in Vietnam by examining the prevalence of mcr-1 in fecal samples from chickens and humans. Detection of mcr-1-carrying bacteria in chicken samples was associated with colistin use and detection in human samples with exposure to mcr-1-positive chickens.

Robinson TP, Bu DP, Carrique-Mas J, Fèvre EM, Gilbert M, Grace D, Hay SI, Jiwakanon J, Kakkar M, Kariuki S et al. 2017. Antibiotic resistance: mitigation opportunities in livestock sector development. Animal, 11 (1), pp. 1-3. | Read more

Nhung NT, Cuong NV, Thwaites G, Carrique-Mas J. 2016. Antimicrobial Usage and Antimicrobial Resistance in Animal Production in Southeast Asia: A Review. Antibiotics (Basel), 5 (4), pp. 37-37. | Show Abstract | Read more

Southeast Asia is an area of great economic dynamism. In recent years, it has experienced a rapid rise in the levels of animal product production and consumption. The region is considered to be a hotspot for infectious diseases and antimicrobial resistance (AMR). We reviewed English-language peer-reviewed publications related to antimicrobial usage (AMU) and AMR in animal production, as well as antimicrobial residues in meat and fish from 2000 to 2016, in the region. There is a paucity of data from most countries and for most bacterial pathogens. Most of the published work relates to non-typhoidal Salmonella (NTS), Escherichia coli (E. coli), and Campylobacter spp. (mainly from Vietnam and Thailand), Enterococcus spp. (Malaysia), and methicillin-resistant Staphylococcus aureus (MRSA) (Thailand). However, most studies used the disk diffusion method for antimicrobial susceptibility testing; breakpoints were interpreted using Clinical Standard Laboratory Institute (CSLI) guidelines. Statistical models integrating data from publications on AMR in NTS and E. coli studies show a higher overall prevalence of AMR in pig isolates, and an increase in levels of AMR over the years. AMU studies (mostly from Vietnam) indicate very high usage levels of most types of antimicrobials, including beta-lactams, aminoglycosides, macrolides, and quinolones. This review summarizes information about genetic determinants of resistance, most of which are transferrable (mostly plasmids and integrons). The data in this review provide a benchmark to help focus research and policies on AMU and AMR in the region.

Trung NV, Nhung HN, Carrique-Mas JJ, Mai HH, Tuyen HT, Campbell J, Nhung NT, Van Minh P, Wagenaar JA, Mai NTN et al. 2016. Colonization of Enteroaggregative Escherichia coli and Shiga toxin-producing Escherichia coli in chickens and humans in southern Vietnam. BMC Microbiol, 16 (1), pp. 208. | Show Abstract | Read more

BACKGROUND: Enteroaggregative (EAEC) and Shiga-toxin producing Escherichia coli (STEC) are a major cause of diarrhea worldwide. E. coli carrying both virulence factors characteristic for EAEC and STEC and producing extended-spectrum beta-lactamase caused severe and protracted disease during an outbreak of E. coli O104:H4 in Europe in 2011. We assessed the opportunities for E. coli carrying the aggR and stx genes to emerge in 'backyard' farms in south-east Asia. RESULTS: Faecal samples collected from 204 chicken farms; 204 farmers and 306 age- and gender-matched individuals not exposed to poultry farming were plated on MacConkey agar plates with and without antimicrobials being supplemented. Sweep samples obtained from MacConkey agar plates without supplemented antimicrobials were screened by multiplex PCR for the detection of the stx1, stx2 and aggR genes. One chicken farm sample each (0.5 %) contained the stx1 and the aggR gene. Eleven (2.4 %) human faecal samples contained the stx1 gene, 2 samples (0.4 %) contained stx2 gene, and 31 (6.8 %) contained the aggR gene. From 46 PCR-positive samples, 205 E. coli isolates were tested for the presence of stx1, stx2, aggR, wzx O104 and fliC H4 genes. None of the isolates simultaneously contained the four genetic markers associated with E. coli O104:H4 epidemic strain (aggR, stx2, wzx O104 and fliC H4 ). Of 34 EAEC, 64.7 % were resistant to 3(rd)-generation cephalosporins. CONCLUSION: These results indicate that in southern Vietnam, the human population is a more likely reservoir of aggR and stx gene carrying E. coli than the chicken population. However, conditions for transmission of isolates and/or genes between human and animal reservoirs resulting in the emergence of highly virulent E. coli strains are still favorable, given the nature of'backyard' farms in Vietnam.

Robinson TP, Bu DP, Carrique-Mas J, Fèvre EM, Gilbert M, Grace D, Hay SI, Jiwakanon J, Kakkar M, Kariuki S et al. 2016. Antibiotic resistance is the quintessential One Health issue. Trans R Soc Trop Med Hyg, 110 (7), pp. 377-380. | Read more

Nguyen NT, Nguyen HM, Nguyen CV, Nguyen TV, Nguyen MT, Thai HQ, Ho MH, Thwaites G, Ngo HT, Baker S, Carrique-Mas J. 2016. Use of Colistin and Other Critical Antimicrobials on Pig and Chicken Farms in Southern Vietnam and Its Association with Resistance in Commensal Escherichia coli Bacteria. Appl Environ Microbiol, 82 (13), pp. 3727-3735. | Show Abstract | Read more

UNLABELLED: Antimicrobial resistance (AMR) is a global health problem, and emerging semi-intensive farming systems in Southeast Asia are major contributors to the AMR burden. We accessed 12 pig and chicken farms at key stages of production in Tien Giang Province, Vietnam, to measure antimicrobial usage and to investigate the prevalence of AMR to five critical antimicrobials (β-lactams, third-generation cephalosporins, quinolones, aminoglycosides, and polymyxins) and their corresponding molecular mechanisms among 180 Escherichia coli isolates. Overall, 94.7 mg (interquartile range [IQR], 65.3 to 151.1) and 563.6 mg (IQR, 398.9 to 943.6) of antimicrobials was used to produce 1 kg (live weight) of chicken and pig, respectively. A median of 3 (out of 8) critical antimicrobials were used on pig farms. E. coli isolates exhibited a high prevalence of resistance to ampicillin (97.8% and 94.4% for chickens and pigs, respectively), ciprofloxacin (73.3% and 21.1%), gentamicin (42.2% and 35.6%), and colistin (22.2% and 24.4%). The prevalence of a recently discovered colistin resistance gene, mcr-1, was 19 to 22% and had strong agreement with phenotypic colistin resistance. We conducted plasmid conjugation experiments with 37 mcr-1 gene-positive E. coli isolates and successfully observed transfer of the gene in 54.0% of isolates through a plasmid of approximately 63 kb, consistent with one recently identified in China. We found no significant correlation between total use of antimicrobials at the farm level and AMR. These data provide additional insight into the role of mcr-1 in colistin resistance on farms and outline the dynamics of phenotypic and genotypic AMR in semi-intensive farming systems in Vietnam. IMPORTANCE: Our study provides accurate baseline information on levels of antimicrobial use, as well as on the dynamics of phenotypic and genotypic resistance for antimicrobials of critical importance among E. coli over the different stages of production in emerging pig and poultry production systems in Vietnam. E. coli isolates showed a high prevalence of resistance (>20%) to critically important antimicrobials, such as colistin, ciprofloxacin, and gentamicin. The underlying genetic mechanisms identified for colistin (the mcr-1 gene) and quinolone (gyrA gene mutations) are likely to play a major role in AMR to those compounds. Conjugation experiments led to the identification of a 63-kb plasmid, similar to one recently identified in China, as the potential carrier of the mcr-1 gene. These results should encourage greater restrictions of such antimicrobials in Southeast Asian farming systems.

Walther BA, Boëte C, Binot A, By Y, Cappelle J, Carrique-Mas J, Chou M, Furey N, Kim S, Lajaunie C et al. 2016. Biodiversity and health: Lessons and recommendations from an interdisciplinary conference to advise Southeast Asian research, society and policy. Infect Genet Evol, 40 pp. 29-46. | Show Abstract | Read more

Southeast Asia is an economic, biodiverse, cultural and disease hotspot. Due to rapid socio-economic and environmental changes, the role of biodiversity and ecosystems for human health ought to be examined and communicated to decision-makers and the public. We therefore summarized the lessons and recommendations from an interdisciplinary conference convened in Cambodia in 2014 to advise Southeast Asian societies on current research efforts, future research needs, and to provide suggestions for improved education, training and science-policy interactions. First, we reviewed several examples of the important role of ecosystems as 'sentinels' in the sense that potentially harmful developments for human health become first apparent in ecosystem components. Other ecosystem services which also benefit human well-being are briefly summarized. Second, we summarized the recommendations of the conference's roundtable discussions and added recent developments in the science-policy interface. The recommendations were organized along five themes: Ethical and legal considerations; implementation of the One Health approach; education, training, and capacity building; future research priorities; and potential science-policy interactions. While the role of biodiversity for human health needs further research, especially for zoonoses and emerging diseases, many direct and indirect benefits to human health are already apparent, but have yet to filter down to the science-policy interface in order to influence legislation and enforcement. Therefore, efforts to strengthen the interface in Southeast Asia should become a high priority in order to strengthen the health and resilience of Southeast Asian societies.

Van Cuong N, Nhung NT, Nghia NH, Mai Hoa NT, Trung NV, Thwaites G, Carrique-Mas J. 2016. Antimicrobial Consumption in Medicated Feeds in Vietnamese Pig and Poultry Production. Ecohealth, 13 (3), pp. 490-498. | Show Abstract | Read more

Antimicrobials are extensively used as growth promoters in animal feeds worldwide, but reliable estimates are lacking. We conducted an internet-based survey of commercial chicken and pig feed products officially approved for sale in Vietnam over the period March-June 2015. Information on the antimicrobial contents in feed products, alongside animal production data, was used to estimate in-feed antimicrobial consumption to produce one kilogram of live animal (chicken, pig), as well as to estimate country-wide antimicrobial consumption through animal feeds. A total of 1462 commercial feed formulations were examined. The survey-adjusted estimated antimicrobial contents were 25.7 and 62.3 mg/kg in chicken and pig feeds, respectively. Overall, it was estimated that 77.4 mg [95% CI 48.1-106.8] and 286.6 mg [95% CI 191.6-418.3] of in-feed antimicrobials were used to raise 1 kg of live chicken and pig, respectively. Bacitracin (15.5% feeds), chlortetracycline (11.4%), and enramycin (10.8%) were the most common antimicrobials present in chicken feed formulations, whereas bacitracin (24.8%), chlortetracycline (23.9%), and florfenicol (17.4%) were the most common in pig feed formulations. Overall, 57% of the total quantitative usage consisted of antimicrobials regarded by WHO of importance for human medicine, including amoxicillin, colistin, tetracyclines, neomycin, lincomycin, and bacitracin. These figures confirm a very high magnitude of in-feed consumption of antimicrobials, especially in pig production. Results from this study should encourage further monitoring of antimicrobials used in animal production, and foster discussion about existing policies on inclusion of antimicrobials in animal feed rations.

Trung NV, Carrique-Mas JJ, Nghia NH, Tu LTP, Mai HH, Tuyen HT, Campbell J, Nhung NT, Nhung HN, Minh PV et al. 2017. Non-Typhoidal Salmonella Colonization in Chickens and Humans in the Mekong Delta of Vietnam. Zoonoses Public Health, 64 (2), pp. 94-99. | Show Abstract | Read more

Salmonellosis is a public health concern in both the developed and developing countries. Although the majority of human non-typhoidal Salmonella enterica (NTS) cases are the result of foodborne infections or person-to-person transmission, NTS infections may also be acquired by environmental and occupational exposure to animals. While a considerable number of studies have investigated the presence of NTS in farm animals and meat/carcasses, very few studies have investigated the risk of NTS colonization in humans as a result of direct animal exposure. We investigated asymptomatic NTS colonization in 204 backyard chicken farms, 204 farmers and 306 matched individuals not exposed to chicken farming, in southern Vietnam. Pooled chicken faeces, collected using boot or handheld swabs on backyard chicken farms, and rectal swabs from human participants were tested. NTS colonization prevalence was 45.6%, 4.4% and 2.6% for chicken farms, farmers and unexposed individuals, respectively. Our study observed a higher prevalence of NTS colonization among chicken farmers (4.4%) compared with age-, sex- and location- matched rural and urban individuals not exposed to chickens (2.9% and 2.0%). A total of 164 chicken NTS strains and 17 human NTS strains were isolated, and 28 serovars were identified. Salmonella Weltevreden was the predominant serovar in both chickens and humans. NTS isolates showed resistance (20-40%) against tetracycline, chloramphenicol, sulfamethoxazole-trimethoprim and ampicillin. Our study reflects the epidemiology of NTS colonization in chickens and humans in the Mekong delta of Vietnam and emphasizes the need of larger, preferably longitudinal studies to study the transmission dynamics of NTS between and within animal and human host populations.

Van Vinh Chau N, Buu Chau L, Desquesnes M, Herder S, Phu Huong Lan N, Campbell JI, Van Cuong N, Yimming B, Chalermwong P, Jittapalapong S et al. 2016. A Clinical and Epidemiological Investigation of the First Reported Human Infection With the Zoonotic Parasite Trypanosoma evansi in Southeast Asia. Clin Infect Dis, 62 (8), pp. 1002-1008. | Show Abstract | Read more

BACKGROUND: Trypanosomais a genus of unicellular parasitic flagellate protozoa.Trypanosoma bruceispecies and Trypanosoma cruziare the major agents of human trypanosomiasis; other Trypanosomaspecies can cause human disease, but are rare. In March 2015, a 38-year-old woman presented to a healthcare facility in southern Vietnam with fever, headache, and arthralgia. Microscopic examination of blood revealed infection with Trypanosoma METHODS: Microscopic observation, polymerase chain reaction (PCR) amplification of blood samples, and serological testing were performed to identify the infecting species. The patient's blood was screened for the trypanocidal protein apolipoprotein L1 (APOL1), and a field investigation was performed to identify the zoonotic source. RESULTS: PCR amplification and serological testing identified the infecting species as Trypanosoma evansi.Despite relapsing 6 weeks after completing amphotericin B therapy, the patient made a complete recovery after 5 weeks of suramin. The patient was found to have 2 wild-type APOL1 alleles and a normal serum APOL1 concentration. After responsive animal sampling in the presumed location of exposure, cattle and/or buffalo were determined to be the most likely source of the infection, with 14 of 30 (47%) animal blood samples testing PCR positive forT. evansi. CONCLUSIONS: We report the first laboratory-confirmed case ofT. evansiin a previously healthy individual without APOL1 deficiency, potentially contracted via a wound while butchering raw beef, and successfully treated with suramin. A linked epidemiological investigation revealed widespread and previously unidentified burden ofT. evansiin local cattle, highlighting the need for surveillance of this infection in animals and the possibility of further human cases.

Cuong NV, Truc VNT, Nhung NT, Thanh TT, Chieu TTB, Hieu TQ, Men NT, Mai HH, Chi HT, Boni MF et al. 2016. Highly Pathogenic Avian Influenza Virus A/H5N1 Infection in Vaccinated Meat Duck Flocks in the Mekong Delta of Vietnam. Transbound Emerg Dis, 63 (2), pp. 127-135. | Show Abstract | Read more

We investigated episodes of suspected highly pathogenic avian influenza (HPAI)-like illness among 12 meat duck flocks in two districts in Tien Giang province (Mekong Delta, Vietnam) in November 2013. In total, duck samples from 8 of 12 farms tested positive for HPAI virus subtype A/haemagglutinin 5 and neuraminidase 1 (H5N1) by real-time RT-PCR. Sequencing results confirmed clade of 2.3.2.1.c as the cause of the outbreaks. Most (7/8) laboratory-confirmed positive flocks had been vaccinated with inactivated HPAI H5N1 clade 2.3.4 vaccines <6 days prior to onset of clinical signs. A review of vaccination data in relation to estimated production in the area suggested that vaccination efforts were biased towards larger flocks and that vaccination coverage was low [21.2% ducks vaccinated with two shots (range by district 7.4-34.9%)]. The low-coverage data, the experimental evidence of lack of cross-protection conferred by the currently used vaccines based on clade 2.3.4 together with the short lifespan of meat duck flocks (60-70 days), suggest that vaccination is not likely to be effective as a tool for control of H5N1 infection in meat duck flocks in the area.

Oude Munnink BB, Phan MVT, van der Hoek L, Kellam P, Cotten M. 2016. Genome Sequences of a Novel Vietnamese Bat Bunyavirus Genome Announcements, 4 (6), pp. e01366-16-e01366-16. | Show Abstract | Read more

© 2016 Oude Munnink et al. To document the viral zoonotic risks in Vietnam, fecal samples were systematically collected from a number of mammals in southern Vietnam and subjected to agnostic deep sequencing. We describe here novel Vietnamese bunyavirus sequences detected in bat feces. The complete L and S segments from 14 viruses were determined.

Oude Munnink BB, Phan MVT, Kellam P, Cotten M. 2016. Complete Genome Characterization of Two Wild-Type Measles Viruses from Vietnamese Infants during the 2014 Outbreak Genome Announcements, 4 (2), pp. e00250-16-e00250-16. | Show Abstract | Read more

© 2016 Oude Munnink et al. A large measles virus outbreak occurred across Vietnam in 2014. We identified and obtained complete measles virus genomes in stool samples collected from two diarrheal pediatric patients in Dong Thap Province. These are the first complete genome sequences of circulating measles viruses in Vietnam during the 2014 measles outbreak.

Lu L, Van Dung N, Bryant JE, Carrique-Mas J, Van Cuong N, Anh PH, Rabaa MA, Baker S, Simmonds P, Woolhouse ME. 2016. Evolution and phylogeographic dissemination of endemic porcine picornaviruses in Vietnam. Virus Evol, 2 (1), pp. vew001. | Show Abstract | Read more

Members of the Picornaviridae are important and often zoonotic viruses responsible for a variety of human and animal diseases. However, the evolution and spatial dissemination of different picornaviruses circulating in domestic animals are not well studied. We examined the rate of evolution and time of origin of porcine enterovirus G (EV-G) and porcine kobuvirus species C lineages (PKV-C) circulating in pig farms in Vietnam and from other countries. We further explored the spatiotemporal spread of EV-G and PKV-C in Southwest Vietnam using phylogeographic models. Multiple types of EV-G are co-circulating in Vietnam. The two dominant EV-G types among isolates from Vietnam (G1 and G6) showed strong phylogenetic clustering. Three clades of PKV-C (PKV-C1-3) represent more recent introductions into Vietnam; PKV-C2 is closely related to PKV-C from Southwest China, indicating possible cross-border dissemination. In addition, high virus lineage migration rates were estimated within four districts in Dong Thap province in Vietnam for both EV-G types (G1, G6) and all PKV-C (C1-3) clades. We found that Chau Thanh district is a primary source of both EV-G and PKV-C clades, consistent with extensive pig trading in and out of the district. Understanding the evolution and spatial dissemination of endemic picornaviruses in pigs may inform future strategies for the surveillance and control of picornaviruses.

Carrique-Mas JJ, Tue NT, Bryant JE, Saylors K, Cuong NV, Hoa NT, An NN, Hien VB, Lao PV, Tu NC et al. 2015. The baseline characteristics and interim analyses of the high-risk sentinel cohort of the Vietnam Initiative on Zoonotic InfectiONS (VIZIONS). Sci Rep, 5 (1), pp. 17965. | Show Abstract | Read more

The Vietnam Initiative for Zoonotic Infections (VIZIONS) includes community-based 'high-risk sentinel cohort' (HRSC) studies investigating individuals at risk of zoonotic infection due to occupational or residential exposure to animals. A total of 852 HRSC members were recruited between March 2013 and August 2014 from three provinces (Ha Noi, Dak Lak, and Dong Thap). The most numerous group (72.8%) corresponded to individuals living on farms, followed by slaughterers (16.3%) and animal health workers (8.5%). Nasal/pharyngeal and rectal swabs were collected from HRSC members at recruitment and after notifying illness. Exposure to exotic animals (including wild pigs, porcupine, monkey, civet, bamboo rat and bat) was highest for the Dak Lak cohort (53.7%), followed by Ha Noi (13.7%) and Dong Thap (4.0%). A total of 26.8% of individuals reported consumption of raw blood over the previous year; 33.6% slaughterers reported no use of protective equipment at work. Over 686 person-years of observation, 213 episodes of suspect infectious disease were notified, equivalent of 0.35 reports per person-year. Responsive samples were collected from animals in the farm cohort. There was noticeable time and space clustering of disease episodes suggesting that the VIZIONS set up is also suitable for the formal epidemiological investigation of disease outbreaks.

Van Dung N, Anh PH, Van Cuong N, Hoa NT, Carrique-Mas J, Hien VB, Sharp C, Rabaa M, Berto A, Campbell J et al. 2016. Large-scale screening and characterization of enteroviruses and kobuviruses infecting pigs in Vietnam. J Gen Virol, 97 (2), pp. 378-388. | Show Abstract | Read more

A recent survey of pigs in Dong Thap province, Vietnam identified a high frequency of enterovirus species G (EV-G) infection (144/198; 72.7%). Amongst these was a plethora of EV-G types (EV-G1, EV-G6 and four new types EV-G8-EV-G11). To better characterize the genetic diversity of EV-G and investigate the possible existence of further circulating types, we performed a larger-scale study on 484 pig and 45 farm-bred boar faecal samples collected in 2012 and 2014, respectively. All samples from the previous and current studies were also screened for kobuviruses. The overall EV infection frequency remained extremely high (395/484; 81.6%), but with comparable detection rates and viral loads between healthy and diarrhoeic pigs; this contrasted with less frequent detection of EV-G in boars (4/45; 8.9%). EV was most frequently detected in pigs ≤ 14 weeks old (∼ 95%) and declined in older pigs. Infections with EV-G1 and EV-G6 were most frequent, whilst less commonly detected types included EV-G3, EV-G4 and EV-G8-EV-G11, and five new types (EV-G12-EV-G16). In contrast, kobuvirus infection frequency was significantly higher in diarrhoeic pigs (40.9 versus 27.6%; P = 0.01). Kobuviruses also showed contrasting epizootiologies and age associations; a higher prevalence was found in boars (42%) compared with domestic pigs (29%), with the highest infection frequency amongst pigs >52 weeks old. Although genetically diverse, all kobuviruses identified belonged to the species Aichivirus C. In summary, this study confirms infection with EV-G was endemic in Vietnamese domestic pigs and exhibits high genetic diversity and extensive inter-type recombination.

Nhung NT, Thuy CT, Trung NV, Campbell J, Baker S, Thwaites G, Hoa NT, Carrique-Mas J. 2015. Induction of Antimicrobial Resistance in Escherichia coli and Non-Typhoidal Salmonella Strains after Adaptation to Disinfectant Commonly Used on Farms in Vietnam. Antibiotics (Basel), 4 (4), pp. 480-494. | Show Abstract | Read more

In Vietnam, commercial disinfectants containing quaternary ammonium compounds (QACs) are commonly used in pig and poultry farms to maintain hygiene during production. We hypothesized that sustained exposure to sub-bactericidal concentrations of QAC-based disinfectants may result in increased levels of antimicrobial resistance (AMR) among Enterobacteriacea due to the increase of efflux pump expression. To test this hypothesis we exposed six antimicrobial-susceptible Escherichia coli (E. coli) and six antimicrobial-susceptible non-typhoidal Salmonella (NTS) isolates to increasing concentrations of a commonly used commercial disinfectant containing a mix of benzalkonium chloride and glutaraldehyde. Over the 12-day experiment, strains exhibited a significant change in their minimum inhibitory concentration (MIC) of the disinfectant product (mean increase of 31% (SD ± 40)) (p = 0.02, paired Wilcoxon test). Increases in MIC for the disinfectant product were strongly correlated with increases in MIC (or decreases in inhibition zone) for all antimicrobials (Pearson's correlation coefficient 0.71-0.83, all p < 0.01). The greatest increases in MIC (or decreases in inhibition zone) were observed for ampicillin, tetracycline, ciprofloxacin, and chloramphenicol, and the smallest for gentamicin, trimethoprim/sulphamethoxazole. The treatment of 155 representative E. coli isolates from farmed and wild animals in the Mekong Delta (Vietnam) with phenyl-arginine beta-naphthylamide (PAβN), a generic efflux pump inhibitor, resulted in reductions in the prevalence of AMR ranging from 0.7% to 3.3% in these organisms, indicating a small contribution of efflux pumps on the observed prevalence of AMR on farms. These results suggest that the mass usage of commercial disinfectants, many of which contain QACs, is potentially a contributing factor on the generation and maintenance of AMR in animal production in Vietnam.

Rabaa MA, Tue NT, Phuc TM, Carrique-Mas J, Saylors K, Cotten M, Bryant JE, Nghia HDT, Cuong NV, Pham HA et al. 2015. The Vietnam Initiative on Zoonotic Infections (VIZIONS): A Strategic Approach to Studying Emerging Zoonotic Infectious Diseases. Ecohealth, 12 (4), pp. 726-735. | Show Abstract | Read more

The effect of newly emerging or re-emerging infectious diseases of zoonotic origin in human populations can be potentially catastrophic, and large-scale investigations of such diseases are highly challenging. The monitoring of emergence events is subject to ascertainment bias, whether at the level of species discovery, emerging disease events, or disease outbreaks in human populations. Disease surveillance is generally performed post hoc, driven by a response to recent events and by the availability of detection and identification technologies. Additionally, the inventory of pathogens that exist in mammalian and other reservoirs is incomplete, and identifying those with the potential to cause disease in humans is rarely possible in advance. A major step in understanding the burden and diversity of zoonotic infections, the local behavioral and demographic risks of infection, and the risk of emergence of these pathogens in human populations is to establish surveillance networks in populations that maintain regular contact with diverse animal populations, and to simultaneously characterize pathogen diversity in human and animal populations. Vietnam has been an epicenter of disease emergence over the last decade, and practices at the human/animal interface may facilitate the likelihood of spillover of zoonotic pathogens into humans. To tackle the scientific issues surrounding the origins and emergence of zoonotic infections in Vietnam, we have established The Vietnam Initiative on Zoonotic Infections (VIZIONS). This countrywide project, in which several international institutions collaborate with Vietnamese organizations, is combining clinical data, epidemiology, high-throughput sequencing, and social sciences to address relevant one-health questions. Here, we describe the primary aims of the project, the infrastructure established to address our scientific questions, and the current status of the project. Our principal objective is to develop an integrated approach to the surveillance of pathogens circulating in both human and animal populations and assess how frequently they are exchanged. This infrastructure will facilitate systematic investigations of pathogen ecology and evolution, enhance understanding of viral cross-species transmission events, and identify relevant risk factors and drivers of zoonotic disease emergence.

Anh PH, Van Cuong N, Son NT, Tue NT, Kosoy M, Woolhouse MEJ, Baker S, Bryant JE, Thwaites G, Carrique-Mas JJ, Rabaa MA. 2015. Diversity of Bartonella spp. in Bats, Southern Vietnam. Emerg Infect Dis, 21 (7), pp. 1266-1267. | Read more

Tu LTP, Hoang NVM, Cuong NV, Campbell J, Bryant JE, Hoa NT, Kiet BT, Thompson C, Duy DT, Phat VV et al. 2015. High levels of contamination and antimicrobial-resistant non-typhoidal Salmonella serovars on pig and poultry farms in the Mekong Delta of Vietnam. Epidemiol Infect, 143 (14), pp. 3074-3086. | Show Abstract | Read more

We investigated the prevalence, diversity, and antimicrobial resistance (AMR) profiles of non-typhoidal Salmonella (NTS) and associated risk factors on 341 pig, chicken, and duck farms in Dong Thap province (Mekong Delta, Vietnam). Sampling was stratified by species, district (four categories), and farm size (three categories). Pooled faeces, collected using boot swabs, were tested using ISO 6575: 2002 (Annex D). Isolates were serogrouped; group B isolates were tested by polymerase chain reaction to detect S. Typhimurium and (monophasic) serovar 4,[5],12:i:- variants. The farm-level adjusted NTS prevalence was 64·7%, 94·3% and 91·3% for chicken, duck and pig farms, respectively. Factors independently associated with NTS were duck farms [odds ratio (OR) 21·2], farm with >50 pigs (OR 11·9), pig farm with 5-50 pigs (OR 4·88) (vs. chickens), and frequent rodent sightings (OR 2·3). Both S. Typhimurium and monophasic S. Typhimurium were more common in duck farms. Isolates had a high prevalence of resistance (77·6%) against tetracycline, moderate resistance (20-30%) against chloramphenicol, sulfamethoxazole-trimethoprim, ampicillin and nalidixic acid, and low resistance (<5%) against ciprofloxacin and third-generation cephalosporins. Multidrug resistance (resistance against ⩾3 classes of antimicrobial) was independently associated with monophasic S. Typhimurium and other group B isolates (excluding S. Typhimurium) and pig farms. The unusually high prevalence of NTS on Mekong Delta farms poses formidable challenges for control.

Nguyen VT, Carrique-Mas JJ, Ngo TH, Ho HM, Ha TT, Campbell JI, Nguyen TN, Hoang NN, Pham VM, Wagenaar JA et al. 2015. Prevalence and risk factors for carriage of antimicrobial-resistant Escherichia coli on household and small-scale chicken farms in the Mekong Delta of Vietnam. J Antimicrob Chemother, 70 (7), pp. 2144-2152. | Show Abstract | Read more

OBJECTIVES: To describe the prevalence of antimicrobial resistance among commensal Escherichia coli isolates on household and small-scale chicken farms, common in southern Vietnam, and to investigate the association of antimicrobial resistance with farming practices and antimicrobial usage. METHODS: We collected data on farming and antimicrobial usage from 208 chicken farms. E. coli was isolated from boot swab samples using MacConkey agar (MA) and MA with ceftazidime, nalidixic acid or gentamicin. Isolates were tested for their susceptibility to 11 antimicrobials and for ESBL production. Risk factor analyses were carried out, using logistic regression, at both the bacterial population and farm levels. RESULTS: E. coli resistant to gentamicin, ciprofloxacin and third-generation cephalosporins was detected on 201 (96.6%), 191 (91.8%) and 77 (37.0%) of the farms, respectively. Of the 895 E. coli isolates, resistance to gentamicin, ciprofloxacin and third-generation cephalosporins was detected in 178 (19.9%), 291 (32.5%) and 29 (3.2%) of the isolates, respectively. Ciprofloxacin resistance was significantly associated with quinolone usage (OR = 2.26) and tetracycline usage (OR = 1.70). ESBL-producing E. coli were associated with farms containing fish ponds (OR = 4.82). CONCLUSIONS: Household and small farms showed frequent antimicrobial usage associated with a high prevalence of resistance to the most commonly used antimicrobials. Given the weak biocontainment, the high prevalence of resistant E. coli could represent a risk to the environment and to humans.

Morris VK, Carrique-Mas JJ, Mueller-Doblies D, Davies RH, Wales AD, Allen VM. 2015. A longitudinal observational study of Salmonella shedding patterns by commercial turkeys during rearing and fattening, showing limitations of some control measures. Br Poult Sci, 56 (1), pp. 48-57. | Show Abstract | Read more

1. The onset and progression of Salmonella infections was investigated in commercial turkey flocks from placement at 1 d old until slaughter in "brood and move" systems using a longitudinal observational approach based on faeces and environmental sampling with subsequent culture of Salmonella. 2. Persistent Salmonella Newport contamination was found within rearing houses and on their external concrete aprons after cleaning and disinfection between crops of heavily shedding young birds. 3. Salmonella shedding was often detected by 5 d of age and the frequency of positive samples peaked at 14-35 d. Thereafter Salmonella isolations declined, especially in the later (fattening) stages. Samples were still Salmonella-positive at low prevalence in half of the intensively sampled houses at slaughter age. 4. A number of management interventions to combat Salmonella infection of flocks, including sourcing policy, competitive exclusion cultures and cleaning and disinfection, were inadequate to prevent flock infection, although improved disinfection on one unit was associated with a delay in the onset of flock infection.

Van Cuong N, Carrique-Mas J, Vo Be H, An NN, Tue NT, Anh NL, Anh PH, Phuc NT, Baker S, Voutilainen L et al. 2015. Rodents and risk in the Mekong Delta of Vietnam: seroprevalence of selected zoonotic viruses in rodents and humans. Vector Borne Zoonotic Dis, 15 (1), pp. 65-72. | Show Abstract | Read more

In the Mekong Delta in southern Vietnam, rats are commonly traded in wet markets and sold live for food consumption. We investigated seroprevalence to selected groups of rodent-borne viruses among human populations with high levels of animal exposure and among co-located rodent populations. The indirect fluorescence antibody test (IFAT) was used to determine seropositivity to representative reference strains of hantaviruses (Dobrava virus [DOBV], Seoul virus [SEOV]), cowpox virus, arenaviruses (lymphocytic choriomeningitis virus [LCMV]), flaviviruses (tick-borne encephalitis virus [TBEV]), and rodent parechoviruses (Ljungan virus), using sera from 245 humans living in Dong Thap Province and 275 rodents representing the five common rodent species sold in wet markets and present in peridomestic and farm settings. Combined seropositivity to DOBV and SEOV among the rodents and humans was 6.9% (19/275) and 3.7% (9/245), respectively; 1.1% (3/275) and 4.5% (11/245) to cowpox virus; 5.4% (15/275) and 47.3% (116/245) for TBEV; and exposure to Ljungan virus was 18.8% (46/245) in humans, but 0% in rodents. Very little seroreactivity was observed to LCMV in either rodents (1/275, 0.4%) or humans (2/245, 0.8%). Molecular screening of rodent liver tissues using consensus primers for flaviviruses did not yield any amplicons, whereas molecular screening of rodent lung tissues for hantavirus yielded one hantavirus sequence (SEOV). In summary, these results indicate low to moderate levels of endemic hantavirus circulation, possible circulation of a flavivirus in rodent reservoirs, and the first available data on human exposures to parechoviruses in Vietnam. Although the current evidence suggests only limited exposure of humans to known rodent-borne diseases, further research is warranted to assess public health implications of the rodent trade.

Tu LTP, Hoang NVM, Cuong NV, Campbell J, Bryant JE, Hoa NT, Kiet BT, Thompson C, Duy DT, Phat VV et al. 2015. High levels of contamination and antimicrobial-resistant non-typhoidal Salmonella serovars on pig and poultry farms in the Mekong Delta of Vietnam Epidemiology and Infection, 143 (14), pp. 3074-3086. | Show Abstract | Read more

Copyright © 2015 Cambridge University Press. We investigated the prevalence, diversity, and antimicrobial resistance (AMR) profiles of non-typhoidal Salmonella (NTS) and associated risk factors on 341 pig, chicken, and duck farms in Dong Thap province (Mekong Delta, Vietnam). Sampling was stratified by species, district (four categories), and farm size (three categories). Pooled faeces, collected using boot swabs, were tested using ISO 6575: 2002 (Annex D). Isolates were serogrouped; group B isolates were tested by polymerase chain reaction to detect S. Typhimurium and (monophasic) serovar 4,[5],12:i:- variants. The farm-level adjusted NTS prevalence was 64.7%, 94.3% and 91.3% for chicken, duck and pig farms, respectively. Factors independently associated with NTS were duck farms [odds ratio (OR) 21.2] , farm with > 50 pigs (OR 11.9), pig farm with 5-50 pigs (OR 4.88) (vs. chickens), and frequent rodent sightings (OR 2.3). Both S. Typhimurium and monophasic S. Typhimurium were more common in duck farms. Isolates had a high prevalence of resistance (77.6%) against tetracycline, moderate resistance (20-30%) against chloramphenicol, sulfamethoxazole-trimethoprim, ampicillin and nalidixic acid, and low resistance ( < 5%) against ciprofloxacin and third-generation cephalosporins. Multidrug resistance (resistance against ≥3 classes of antimicrobial) was independently associated with monophasic S. Typhimurium and other group B isolates (excluding S. Typhimurium) and pig farms. The unusually high prevalence of NTS on Mekong Delta farms poses formidable challenges for control.

Loan HK, Cuong NV, Takhampunya R, Klangthong K, Osikowicz L, Kiet BT, Campbell J, Bryant J, Promstaporn S, Kosoy M et al. 2015. Bartonella species and trombiculid mites of rats from the Mekong Delta of Vietnam. Vector Borne Zoonotic Dis, 15 (1), pp. 40-47. | Show Abstract | Read more

A survey of Bartonella spp. from 275 rats purchased in food markets (n=150) and trapped in different ecosystems (rice field, forest, and animal farms) (n=125) was carried out during October, 2012-March, 2013, in the Mekong Delta of Vietnam. The overall Bartonella spp. prevalence detected by culture and PCR in blood was 14.9% (10.7-19.1%), the highest corresponding to Rattus tanezumi (49.2%), followed by Rattus norvegicus (20.7%). Trapped rats were also investigated for the presence and type of chiggers (larvae of trombiculid mites), and Bartonella spp. were investigated on chigger pools collected from each rat by RT-PCR. A total of five Bartonella spp. were identified in rats, three of which (B. elizabethae, B. rattimassiliensis, and B. tribocorum) are known zoonotic pathogens. Among trapped rats, factors independently associated with increased prevalence of Bartonella spp. included: (1) Rat species (R. tanezumi); (2) the number of Trombiculini-Blankaartia and Schoengastiini-Ascoschoengastia mites found on rats; and (3) the habitat of the rat (i.e., forest/fields vs. animal farms). The prevalence of Bartonella infection among chiggers from Bartonella spp.-positive R. tanezumi rats was 5/25 (25%), compared with 1/27 (3.7%) among Bartonella spp.-negative R. tanezumi rats (relative risk [RR]=5.4, 95% confidence interval [CI] 0.68-43.09). The finding of Bartonella spp.-positive chiggers on Bartonella spp.-negative rats is strongly suggestive of a transovarial transmission cycle. Rats are ubiquitous in areas of human activity and farms in the Mekong Delta; in addition, trapping and trading of rats for food is common. To correctly assess the human risks due to rat trapping, marketing, and carcass dressing, further studies are needed to establish the routes of transmission and cycle of infection. The widespread presence of these zoonotic pathogens in rats and the abundance of human-rat interactions suggest that surveillance efforts should be enhanced to detect any human cases of Bartonella infection that may arise.

Carrique-Mas JJ, Trung NV, Hoa NT, Mai HH, Thanh TH, Campbell JI, Wagenaar JA, Hardon A, Hieu TQ, Schultsz C. 2015. Antimicrobial usage in chicken production in the Mekong Delta of Vietnam. Zoonoses Public Health, 62 Suppl 1 (s1), pp. 70-78. | Show Abstract | Read more

Antimicrobials are used extensively in chicken production in Vietnam, but to date no quantitative data are available. A 2012-2013 survey of 208 chicken farms in Tien Giang province, stratified by size (10-200 chickens; >200-2000), was carried out to describe and quantify the use of antibacterial antimicrobials (usage per week per chicken and usage per 1000 chickens produced) in the Mekong Delta and to investigate factors associated with usage. Twenty-eight types of antimicrobial belonging to 10 classes were reported. Sixty-three per cent of all commercial formulations contained at least two antimicrobials. On 84% occasions, antimicrobials were administered with a prophylactic purpose. The overall adjusted quantities of antimicrobials used/week/chicken and per 1000 chickens produced (g) were 26.36 mg (SE ± 3.54) and 690.4 g (SE ± 203.6), respectively. Polypeptides, tetracyclines, penicillins and aminoglycosides were the antimicrobials used by most farms (18.6% farms, 17.5%, 11.3% and 10.1% farms, respectively), whereas penicillins, lincosamides, quinolones, and sulphonamides/trimethoprim were quantitatively the most used compounds (8.27, 5.2, 3.16 and 2.78 mg per week per chicken, respectively). Factors statistically associated with higher levels of usage (per week per chicken) were meat farms (OR = 1.40) and farms run by a male farmer (OR = 2.0). All-in-all-out farming systems (correlated with medium farms) were associated with reduced levels of antimicrobial usage (OR = 0.68). Usage levels to produced meat chickens were considerably higher than those reported in European countries. This should trigger the implementation of surveillance programmes to monitor sales of antimicrobials that should contribute to the rational administration of antimicrobials in order to preserve the efficacy of existing antimicrobials in Vietnam.

Nhung NT, Cuong NV, Campbell J, Hoa NT, Bryant JE, Truc VNT, Kiet BT, Jombart T, Trung NV, Hien VB et al. 2015. High levels of antimicrobial resistance among escherichia coli isolates from livestock farms and synanthropic rats and shrews in the Mekong Delta of Vietnam. Appl Environ Microbiol, 81 (3), pp. 812-820. | Show Abstract | Read more

In Mekong Delta farms (Vietnam), antimicrobials are extensively used, but limited data are available on levels of antimicrobial resistance (AMR) among Escherichia coli isolates. We performed a structured survey of AMR in E. coli isolates (n = 434) from 90 pig, chicken, and duck farms. The results were compared with AMR among E. coli isolates (n = 234) from 66 small wild animals (rats and shrews) trapped on farms and in forests and rice fields. The isolates were susceptibility tested against eight antimicrobials. E. coli isolates from farmed animals were resistant to a median of 4 (interquartile range [IQR], 3 to 6) antimicrobials versus 1 (IQR, 1 to 2) among wild mammal isolates (P < 0.001). The prevalences of AMR among farmed species isolates (versus wild animals) were as follows: tetracycline, 84.7% (versus 25.6%); ampicillin, 78.9% (versus 85.9%); trimethoprim-sulfamethoxazole, 52.1% (versus 18.8%); chloramphenicol, 39.9% (versus 22.5%); amoxicillin-clavulanic acid, 36.6% (versus 34.5%); and ciprofloxacin, 24.9% (versus 7.3%). The prevalence of multidrug resistance (MDR) (resistance against three or more antimicrobial classes) among pig isolates was 86.7% compared to 66.9 to 72.7% among poultry isolates. After adjusting for host species, MDR was ∼8 times greater among isolates from wild mammals trapped on farms than among those trapped in forests/rice fields (P < 0.001). Isolates were assigned to unique profiles representing their combinations of susceptibility results. Multivariable analysis of variance indicated that AMR profiles from wild mammals trapped on farms and those from domestic animals were more alike (R(2) range, 0.14 to 0.30) than E. coli isolates from domestic animals and mammals trapped in the wild (R(2) range, 0.25 to 0.45). The results strongly suggest that AMR on farms is a key driver of environmental AMR in the Mekong Delta.

Harrison JW, Dung TTN, Siddiqui F, Korbrisate S, Bukhari H, Tra MPV, Hoang NVM, Carrique-Mas J, Bryant J, Campbell JI et al. 2014. Identification of possible virulence marker from Campylobacter jejuni isolates. Emerg Infect Dis, 20 (6), pp. 1026-1029. | Show Abstract | Read more

A novel protein translocation system, the type-6 secretion system (T6SS), may play a role in virulence of Campylobacter jejuni. We investigated 181 C. jejuni isolates from humans, chickens, and environmental sources in Vietnam, Thailand, Pakistan, and the United Kingdom for T6SS. The marker was most prevalent in human and chicken isolates from Vietnam.

Pham HA, Carrique-Mas JJ, Nguyen VC, Ngo TH, Nguyet LA, Do TD, Vo BH, Phan VTM, Rabaa MA, Farrar J et al. 2014. The prevalence and genetic diversity of group A rotaviruses on pig farms in the Mekong Delta region of Vietnam. Vet Microbiol, 170 (3-4), pp. 258-265. | Show Abstract | Read more

Group A rotaviruses (ARoVs) are a common cause of severe diarrhea among children worldwide and the cause of approximately 45% of pediatric hospitalizations for acute diarrhea in Vietnam. ARoVs are known to cause significant economic losses to livestock producers by reducing growth performance and production efficiencies, however little is known about the implications of asymptomatic endemic circulation of ARoV. We aimed to determine the prevalence and predominant circulating genotypes of ARoVs on pig farms in a southern province of Vietnam. We found overall animal-level and farm-level prevalence of 32.7% (239/730) and 74% (77/104), respectively, and identified six different G types and 4 P types in various combinations (G2, G3, G4, G5, G9, G11 and P[6], P[13], P[23], and P[34]). There was no significant association between ARoV infection and clinical disease in pigs, suggesting that endemic asymptomatic circulation of ARoV may complicate rotavirus disease attribution during outbreaks of diarrhea in swine. Sequence analysis of the detected ARoVs suggested homology to recent human clinical cases and extensive genetic diversity. The epidemiological relevance of these findings for veterinary practitioners and to ongoing pediatric ARoV vaccine initiatives in Vietnam merits further study.

Mueller-Doblies D, Carrique-Mas JJ, Davies RH. 2014. A study of the dynamics of Salmonella infection in turkey breeding, rearing and finishing houses with special reference to elimination, persistence and introduction of Salmonella. Avian Pathol, 43 (2), pp. 146-154. | Show Abstract | Read more

In this descriptive study, the dynamics of Salmonella infection of turkey flocks were investigated by repeated sampling of houses where Salmonella had been identified. The aim of the study was to identify the most common scenarios involved in elimination, persistence and introduction of Salmonella in the different branches of the turkey industry. Sixty-two houses on 34 turkey farms (comprising breeding, rearing and finishing farms) were sampled longitudinally, starting with the identification of a positive flock. A total of 117 follow-on flocks were tested and cleaning and disinfection (C&D) was assessed during 66 post-C&D visits. A total of 155 incidents (clearance, persistence or introduction of Salmonella) were recorded. Persistence was seen in 35.5% of incidents and was seen more frequently in breeding and rearing houses compared with finishing houses. Most persistence incidents were the result of insufficient C&D. Clearance was seen in 40% of incidents and was more often observed in finishing houses than in breeding or rearing houses. Introduction was seen in 24.5% of incidents and was more common in breeding and finishing flocks than in rearing flocks. Contamination of a house with Salmonella Typhimurium was more likely to be cleared compared with other serovars. The total number of positive samples found at a post-C&D visit was correlated with the probability of carry-over of infection, whereas the location of the positive samples seemed to be less important. Our highly sensitive post-C&D sampling method allowed us to predict a negative follow-on flock in most cases.

Cuong NV, Carrique-Mas J, Thu HTV, Hien ND, Hoa NT, Nguyet LA, Anh PH, Bryant JE. 2014. Serological and virological surveillance for porcine reproductive and respiratory syndrome virus, porcine circovirus type 2, and influenza A viruses among smallholder swine farms of the Mekong Delta, Vietnam JOURNAL OF SWINE HEALTH AND PRODUCTION, 22 (5), pp. 224-231. | Show Abstract

Objectives: To evaluate the feasibility and utility of oral-luids collection for surveillance of porcine viruses in the Mekong Delta, Vietnam, and to establish baseline serological and virological prevalence estimates for porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2(PCV2), and influenza A virus (IAV) among smallholder farms. Materials and methods: Paired serum and oral-luids samples from 68 farms (sows, boars, weaners, and growers) were tested during 2011 by reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay for PRRSV, PCV2, and IAV. Results: Low numbers of PRRSV-positive and IAV-positive pigs were detected (1.6% PRRSV viremic, two of 124; 0.8% IAV in oral luids, one of 124). However, PCV2 detection rates were high in both serum and oral fluids (54.8% and 61.3%, respectively). Overall proportions of pigs seropositive for IAV and PRRSV were 37.9% and 33.9%, respectively. Proportions of pigs seropositive for PRRSV were 48.6% (17 of 35) and 12.1% (four of 33) on vaccinated and unvaccinated farms, respectively. Oral luids and serum samples yielded comparable prevalence estimates for molecular detection of PCV2, and detected one sample PCR-positive for hemagglutinin of influenza A/H1N1/pdm09. There was no evidence of PRRSV shedding in oral luids. Implications: Antibody prevalence estimates based on testing oral luids may prov ide an acceptable and useful surrogate for testing serum in future field studies if optimized assays are employed.

Hong Anh P, Carrique-Mas JJ, Van Cuong N, Hoa NT, Lam Anh N, Duy DT, Hien VB, Vu Tra My P, Rabaa MA, Farrar J et al. 2014. The prevalence and genetic diversity of group A rotaviruses on pig farms in the Mekong Delta region of Vietnam Veterinary Microbiology, 170 (3-4), pp. 258-265. | Show Abstract | Read more

Group A rotaviruses (ARoVs) are a common cause of severe diarrhea among children worldwide and the cause of approximately 45% of pediatric hospitalizations for acute diarrhea in Vietnam. ARoVs are known to cause significant economic losses to livestock producers by reducing growth performance and production efficiencies, however little is known about the implications of asymptomatic endemic circulation of ARoV. We aimed to determine the prevalence and predominant circulating genotypes of ARoVs on pig farms in a southern province of Vietnam. We found overall animal-level and farm-level prevalence of 32.7% (239/730) and 74% (77/104), respectively, and identified six different G types and 4 P types in various combinations (G2, G3, G4, G5, G9, G11 and P[6], P[13] , P[23], and P[34] ). There was no significant association between ARoV infection and clinical disease in pigs, suggesting that endemic asymptomatic circulation of ARoV may complicate rotavirus disease attribution during outbreaks of diarrhea in swine. Sequence analysis of the detected ARoVs suggested homology to recent human clinical cases and extensive genetic diversity. The epidemiological relevance of these findings for veterinary practitioners and to ongoing pediatric ARoV vaccine initiatives in Vietnam merits further study. © 2014 The Authors.

Van Dung N, Anh PH, Van Cuong N, Hoa NT, Carrique-Mas J, Hien VB, Campbell J, Baker S, Farrar J, Woolhouse ME et al. 2014. Prevalence, genetic diversity and recombination of species G enteroviruses infecting pigs in Vietnam. J Gen Virol, 95 (Pt 3), pp. 549-556. | Show Abstract | Read more

Picornaviruses infecting pigs, described for many years as 'porcine enteroviruses', have recently been recognized as distinct viruses within three distinct genera (Teschovirus, Sapelovirus and Enterovirus). To better characterize the epidemiology and genetic diversity of members of the Enterovirus genus, faecal samples from pigs from four provinces in Vietnam were screened by PCR using conserved enterovirus (EV)-specific primers from the 5' untranslated region (5' UTR). High rates of infection were recorded in pigs on all farms, with detection frequencies of approximately 90% in recently weaned pigs but declining to 40% in those aged over 1 year. No differences in EV detection rates were observed between pigs with and without diarrhoea [74% (n = 70) compared with 72% (n = 128)]. Genetic analysis of consensus VP4/VP2 and VP1 sequences amplified from a subset of EV-infected pigs identified species G EVs in all samples. Among these, VP1 sequence comparisons identified six type 1 and seven type 6 variants, while four further VP1 sequences failed to group with any previously identified EV-G types. These have now been formally assigned as EV-G types 8-11 by the Picornavirus Study Group. Comparison of VP1, VP4/VP2, 3D(pol) and 5' UTRs of study samples and those available on public databases showed frequent, bootstrap-supported differences in their phylogenies indicative of extensive within-species recombination between genome regions. In summary, we identified extremely high frequencies of infection with EV-G in pigs in Vietnam, substantial genetic diversity and recombination within the species, and evidence for a much larger number of circulating EV-G types than currently described.

Carrique-Mas JJ, Bryant JE. 2013. A review of foodborne bacterial and parasitic zoonoses in Vietnam EcoHealth, 10 (4), pp. 465-489. | Show Abstract | Read more

Vietnam has experienced unprecedented economic and social development in recent years, and the livestock sector is undergoing significant transformations. Although food animal production is still dominated by small-scale 'backyard' enterprises with mixed crop-livestock or livestock-aquatic systems, there is a trend towards more intensive and vertically integrated operations. Changes in animal production, processing and distribution networks for meat and animal products, and the shift from wet markets to supermarkets will undoubtedly impact food safety risks in Vietnam in unforeseen and complex ways. Here, we review the available published literature on bacterial and parasitic foodborne zoonoses (FBZ) in Vietnam. We report on clinical disease burden and pathogen prevalence in animal reservoirs for a number of important FBZ, and outline opportunities for future research. © 2013 International Association for Ecology and Health.

Jones EM, Snow LC, Carrique-Mas JJ, Gosling RJ, Clouting C, Davies RH. 2013. Risk factors for antimicrobial resistance in Escherichia coli found in GB turkey flocks Veterinary Record, 173 (17), pp. 422-422. | Read more

Carrique-Mas JJ, Bryant JE. 2013. A review of foodborne bacterial and parasitic zoonoses in Vietnam. Ecohealth, 10 (4), pp. 465-489. | Show Abstract | Read more

Vietnam has experienced unprecedented economic and social development in recent years, and the livestock sector is undergoing significant transformations. Although food animal production is still dominated by small-scale 'backyard' enterprises with mixed crop-livestock or livestock-aquatic systems, there is a trend towards more intensive and vertically integrated operations. Changes in animal production, processing and distribution networks for meat and animal products, and the shift from wet markets to supermarkets will undoubtedly impact food safety risks in Vietnam in unforeseen and complex ways. Here, we review the available published literature on bacterial and parasitic foodborne zoonoses (FBZ) in Vietnam. We report on clinical disease burden and pathogen prevalence in animal reservoirs for a number of important FBZ, and outline opportunities for future research.

Carrique-Mas JJ, Bryant JE, Cuong NV, Hoang NVM, Campbell J, Hoang NV, Dung TTN, Duy DT, Hoa NT, Thompson C et al. 2014. An epidemiological investigation of Campylobacter in pig and poultry farms in the Mekong delta of Vietnam. Epidemiol Infect, 142 (7), pp. 1425-1436. | Show Abstract | Read more

Campylobacter are zoonotic pathogens commonly associated with gastroenteritis. To assess the relevance of Campylobacter in Vietnam, an economically transitioning country in SE Asia, we conducted a survey of 343 pig and poultry farms in the Mekong delta, a region characterized by mixed species farming with limited biosecurity. The animal-level prevalence of Campylobacter was 31·9%, 23·9% and 53·7% for chickens, ducks and pigs, respectively. C. jejuni was predominant in all three host species, with the highest prevalence in pigs in high-density production areas. Campylobacter isolates demonstrated high levels of antimicrobial resistance (21% and 100% resistance against ciprofloxacin and erythromycin, respectively). Multilocus sequence type genotyping showed a high level of genetic diversity within C. jejuni, and predicted C. coli inter-species transmission. We suggest that on-going intensification of animal production systems, limited biosecurity, and increased urbanization in Vietnam is likely to result in Campylobacter becoming an increasingly significant cause of human diarrhoeal infections in coming years.

Wagenaar JA, Hendriksen RS, Carrique-Mas J. 2013. Practical considerations of surveillance of Salmonella serovars other than Enteritidis and Typhimurium. Rev Sci Tech, 32 (2), pp. 509-519. | Show Abstract | Read more

Non-typhoid Salmonella serovars other than Salmonella enterica serovars S. Enteritidis (SE) and S.Typhimurium (ST) are isolated throughout the world with huge variations in prevalence. Besides the more generally occurring serovars, such as S. Infantis and S. Hadar, there are many examples of serovars that are principally reported from the regions and are most probably associated with local reservoirs. In most countries of the world, no formal surveillance systems for human salmonellosis are in place and data are limited to ad hoc studies. Data on animals, food and animal feed are even more scarce. The identification of non-SE/ST serovars may be hampered by a lack of experience in serotyping and the availability of quality-assured antisera. Subtyping Salmonella remains important to identify sources of human infections and to target interventions and control measurements. However, in the future, there will be an increasing use of culture-independent diagnostic assays, with the consequence that epidemiological subtyping and antimicrobial susceptibility data will no longer be generated. The validation of these assays for all serovars, particularly the rare ones, needs attention. Although current subtyping based on the Kauffmann-White scheme is well established, and has been shown to be robust, a new generation of subtyping methods will replace it in the near future.

Phan MQ, Henry W, Bui CB, DO DH, Hoang NV, Thu NT, Nguyen TT, LE TD, Diep TQ, Inui K et al. 2013. Detection of HPAI H5N1 viruses in ducks sampled from live bird markets in Vietnam. Epidemiol Infect, 141 (3), pp. 601-611. | Show Abstract | Read more

In Vietnam, highly pathogenic avian influenza (HPAI) H5N1 infections in poultry often occur without concomitant clinical signs and outbreaks are not consistently reported. Live bird markets represent a convenient site for surveillance that does not rely on farmers' notifications. Two H5N1 surveys were conducted at live bird markets/slaughter points in 39 districts (five provinces) in the Red River, Mekong delta, and central Vietnam during January and May 2011. Oropharyngeal and rectal swab samples from 12 480 ducks were tested for H5N1 by reverse transcription-polymerase chain reaction in pools of five. Traders and stallholders were interviewed using standardized questionnaires; 3·3% of pools tested positive. The highest prevalence (6·6%) corresponded to the Mekong delta, and no H5N1 was detected in the two Red River provinces. The surveys identified key risk behaviours of traders and stallholders. It is recommended that market surveys are implemented over time as a tool to evaluate progress in HPAI control in Vietnam.

Green LE, Carrique-Mas JJ, Mason SA, Medley GF. 2012. Patterns of delayed detection and persistence of bovine tuberculosis in confirmed and unconfirmed herd breakdowns in cattle and cattle herds in Great Britain. Prev Vet Med, 106 (3-4), pp. 266-274. | Show Abstract | Read more

Approximately 1500/6000 cattle farms that were depopulated during the foot and mouth epidemic in GB in 2001 had been repopulated and subjected to two unrestricted (herd considered free from bovine tuberculosis (bTB)) herd tests. Factors associated with herd breakdown(s) (HBD) and individual cattle reactor status at the second test were investigated. There were 96 HBD in total, with a 3-fold increased risk of HBD in herds that had had a HBD at the first test after restocking. Two mixed effect models were used to investigate factors associated with 324/246,060 reactor cattle at the second bTB test; 228 reactors were at confirmed HBD and 96 at unconfirmed HBD; 253 (79%) reactors at the second test were present and test negative at the first test. In confirmed HBD, the odds of cattle reacting were higher if the restocked farm had a history of bTB before 2001 and if the source and restocked farms were high frequency tested (HFT) farms (routine bTB tests at ≥1 per 2 years). Reacting cattle were more likely to have been born on the restocked farm before the first test after FMD and less likely to have been purchased from a low frequency tested (LFT) farm (routine bTB tests at 3-4 year intervals) after the first test compared with a baseline of cattle purchased from a LFT farm before the first test. Unconfirmed HBD at the second test was more likely when the first test was a confirmed HBD and when there was a history of bTB in the restocked farm. In contrast to confirmed HBD, cattle purchased from a LFT farm after the first test were at increased risk of reacting at an unconfirmed HBD at the second test. We conclude that a farm history of bTB suggests persistence of bTB on the farm. Confirmed tests indicate exposure to bTB for some time indicated by the increased risk from HFT source and restocked farms and a farm history of bTB. The risks for reactors are related to the farm and herd and duration of exposure to these risks. Therefore, the spread of bTB to naïve herds would be reduced if farmers only introduced cattle known not to have been in herds and on farms exposed to bTB. Management of bTB on farms with bTB is complicated because there is undisclosed infection in cattle and environmental contamination.

Green LE, Carrique-Mas JJ, Mason SA, Medley GF. 2012. Patterns of delayed detection and persistence of bovine tuberculosis in confirmed and unconfirmed herd breakdowns in cattle and cattle herds in Great Britain Preventive Veterinary Medicine, 106 (3-4), pp. 266-274. | Show Abstract | Read more

Approximately 1500/6000 cattle farms that were depopulated during the foot and mouth epidemic in GB in 2001 had been repopulated and subjected to two unrestricted (herd considered free from bovine tuberculosis (bTB)) herd tests. Factors associated with herd breakdown(s) (HBD) and individual cattle reactor status at the second test were investigated. There were 96 HBD in total, with a 3-fold increased risk of HBD in herds that had had a HBD at the first test after restocking. Two mixed effect models were used to investigate factors associated with 324/246,060 reactor cattle at the second bTB test; 228 reactors were at confirmed HBD and 96 at unconfirmed HBD; 253 (79%) reactors at the second test were present and test negative at the first test. In confirmed HBD, the odds of cattle reacting were higher if the restocked farm had a history of bTB before 2001 and if the source and restocked farms were high frequency tested (HFT) farms (routine bTB tests at ≥1 per 2 years). Reacting cattle were more likely to have been born on the restocked farm before the first test after FMD and less likely to have been purchased from a low frequency tested (LFT) farm (routine bTB tests at 3-4 year intervals) after the first test compared with a baseline of cattle purchased from a LFT farm before the first test. Unconfirmed HBD at the second test was more likely when the first test was a confirmed HBD and when there was a history of bTB in the restocked farm. In contrast to confirmed HBD, cattle purchased from a LFT farm after the first test were at increased risk of reacting at an unconfirmed HBD at the second test.We conclude that a farm history of bTB suggests persistence of bTB on the farm. Confirmed tests indicate exposure to bTB for some time indicated by the increased risk from HFT source and restocked farms and a farm history of bTB. The risks for reactors are related to the farm and herd and duration of exposure to these risks. Therefore, the spread of bTB to naïve herds would be reduced if farmers only introduced cattle known not to have been in herds and on farms exposed to bTB. Management of bTB on farms with bTB is complicated because there is undisclosed infection in cattle and environmental contamination. © 2012 Elsevier B.V.

Snow LC, Davies RH, Christiansen KH, Carrique-Mas JJ, Cook AJC, Evans SJ. 2011. Survey of Salmonella prevalence on commercial turkey breeding and fattening farms in the UK in 2006 to 2007. Vet Rec, 169 (19), pp. 493. | Show Abstract | Read more

A total of 29 breeding turkey holdings and 317 fattening turkey holdings were sampled between October 2006 and September 2007 in order to establish the baseline prevalence of Salmonella in turkeys in the UK. The weighted holding level Salmonella prevalence was found to be 20.1 per cent (95 per cent confidence interval [CI] 8.6 to 40.3 per cent) in breeding turkeys and 37.7 per cent (95 per cent CI 33.4 to 42.3 per cent) in fattening turkeys. For breeding turkeys, a weighted flock-level prevalence, as more than one flock per holding was sampled, was estimated at 7.1 per cent (95 per cent CI 3.2 to 14.8 per cent). A total of 13 different serovars were identified in the survey. The most frequent serovar in both turkey flock classes was Salmonella Kottbus, which was found on two breeding holdings and 63 of the fattening holdings giving weighted prevalences of 10.4 per cent (95 per cent CI 2.6 to 34.1 per cent) and 23.0 per cent (95 per cent CI 19.3 to 27.3 per cent), respectively. On breeding holdings, a single isolate of Salmonella Typhimurium, identified as DT12 (weighted prevalence 3.5 per cent [95 per cent CI 0.7 to 15.8 per cent] [holding], 0.7 per cent [95 per cent CI 0.1 to 3.7 per cent] [flock)], was found. On fattening holdings, there were 55 isolates of S Typhimurium from 16 holdings, giving a weighted prevalence of this serovar of 5.4 per cent (95 per cent CI 3.6 to 8.0 per cent). There were no isolates of Salmonella serovars Enteritidis, Hadar, Infantis or Virchow.

Arnold ME, Carrique-Mas JJ, McLaren I, Davies RH. 2011. A comparison of pooled and individual bird sampling for detection of Salmonella in commercial egg laying flocks. Prev Vet Med, 99 (2-4), pp. 176-184. | Show Abstract | Read more

The objective of this study was to determine the sensitivity of culturing pooled samples containing varying numbers of individual droppings for detection of Salmonella in commercial egg-laying flocks relative to the within-flock prevalence. A laboratory experiment was performed to directly measure the effect of diluting positive with negative faeces on the sensitivity of detection, and thus provide priors for a Bayesian model of pooled sampling. Pooled samples made up of different numbers of individual faecal droppings were collected from 20 flocks, and in addition bulked faeces and dust were also sampled using an in-house method that involved collecting 10 dust and 10 faeces samples into jars with buffered peptone water. The results from these flocks were analysed using Bayesian methods for diagnostic test evaluation in the absence of a gold standard, and the sensitivity of each pooled sample type was estimated relative to the within-flock prevalence. The sensitivity of pooled samples depended on the within-flock prevalence, and increased as the prevalence increased. The sensitivity of pooled sampling tended to increase with the number of droppings in the pool, and overall there was a higher proportion of positive samples from the pools of 20, 60 and the in-house faeces pooling method compared to the pools of 10, 5 and the individual droppings. Dust samples were more sensitive than the faeces samples, and so the inclusion of dust in sampling schemes is recommended.

Danguy des Déserts J, Davies RH, Vaughan K, McLaren I, Canning P, Wintrip A, Mueller-Doblies D, Carrique-Mas JJ. 2011. A longitudinal study of Salmonella infection in different types of turkey flocks in Great Britain. Zoonoses Public Health, 58 (3), pp. 200-208. | Show Abstract | Read more

Salmonella is, after Campylobacter, the most reported zoonotic pathogen in the EU. Poultry are a common source of infection to humans, and turkey flocks are commonly colonized with the organism. We investigated the prevalence and risk factors of Salmonella infection in 179 houses in 60 holdings representative of turkey meat and breeder production in Great Britain. From each holding, up to four houses were chosen, and two consecutive flocks per house were sampled/tested for Salmonella to investigate the persistence, elimination and introduction of Salmonella in consecutive crops. At the first sampling, the overall flock-level Salmonella prevalence was 32.8% and 8.9% for meat and breeding flocks respectively. There was a higher prevalence of Salmonella in flocks in the rearing stage than in the fattening and breeding stages. At the first sampling, the flock-level prevalence of Salmonella was 26.8% (95% CI: 20.7-33.7%), while the prevalence level in the subsequent flock was 20.5% (95% CI: 13.6-29.7%). No houses were positive for any of the EU-regulated serovars. The most commonly encountered serovars were S. Kottbus and S. Kedougou. Carry-over of infection was observed in 44.8% of the positive houses, and introduction of new infection occurred in 8.4% of houses. Data from the questionnaires and auditing of all holdings and houses were combined and used to calculate adjusted farm- and house-adjusted risk factors. Significant risk factors were feed from a source other than a national compounder (OR = 2.4), feeder type other than pan feeders (OR = 2.4) and hygiene practices other than terminal cleaning and disinfection using power-washing with sanitizer and anteroom with boot change (OR = 2.8). The study discusses the main challenges currently faced by the industry to control Salmonella in turkey production.

Carrique-Mas JJ, Willmington JA, Papadopoulou C, Watson EN, Davies RH. 2010. Salmonella infection in cattle in Great Britain, 2003 to 2008. Vet Rec, 167 (15), pp. 560-565. | Show Abstract | Read more

Surveillance data for clinical disease in cattle in Great Britain due to Salmonella infections were analysed for the period 2003 to 2008 in order to describe seasonality and to investigate possible associations between Salmonella diagnoses and other variables such as region, climate, age and production type. A clear seasonal pattern was shown for Salmonella infection, coinciding with the second half of the year. The incidence of Salmonella Dublin and Salmonella Typhimurium was highest in the west of the country, which has the greatest cattle density, but this was not a feature of diagnoses with other serovars. Abortion was a more common clinical sign of S Dublin infections, but was relatively unusual in the case of S Typhimurium. The observed clinical picture and age of affected animals were largely determined by the seasonality of dairy cattle calving in Great Britain.

Arnold ME, Carrique-Mas JJ, Davies RH. 2010. Sensitivity of environmental sampling methods for detecting Salmonella Enteritidis in commercial laying flocks relative to the within-flock prevalence. Epidemiol Infect, 138 (3), pp. 330-339. | Show Abstract | Read more

The objective of this study was to estimate the sensitivity of three different sampling/testing methodologies for the detection of Salmonella Enteritidis in commercial egg-laying flocks relative to the within-flock prevalence. The following methods were compared on 21 farms: (1) The European Union (EU) baseline survey method (five faecal and two dust samples); (2) an in-house method that involved collecting 10 dust and 10 faecal samples into jars with buffered peptone water, and (3) a method involving single samples of pooled faeces and dust that has been adopted as a monitoring method for the National Control Programme across the EU (the NCP method). Testing of individual bird ovaries/oviduct and caeca was carried out on each flock, and the sensitivity of each sampling method was estimated relative to the within-flock prevalence using Bayesian methods. Results showed that the sensitivity of all the sampling methods increased as the within-flock prevalence increased, and that all were more efficient than individual bird sampling for detection of S. Enteritidis in commercial flocks. The in-house method was the most sensitive of the methods compared, with a 98% power to detect a 0.1% prevalence, and the NCP method the least sensitive, with a 93% power to detect a prevalence of 20%.

Wales AD, Carrique-Mas JJ, Rankin M, Bell B, Thind BB, Davies RH. 2010. Review of the carriage of zoonotic bacteria by arthropods, with special reference to Salmonella in mites, flies and litter beetles. Zoonoses Public Health, 57 (5), pp. 299-314. | Show Abstract | Read more

This systematic review considers the relationship between arthropods commonly found in and around livestock premises and zoonotic bacteria. The principal focus is upon insects and arachnids on poultry units, where houses, litter and manure provide good conditions for the growth, multiplication and protection of flies, beetles and mites, and where zoonotic pathogens such as Salmonella and Campylobacter are prevalent. Other members of the Enterobacteriaceae and the taxa Clostridium, Helicobacter, Erysipelas and Chlamydiaceae are also discussed. Salmonella is widely distributed in the flies of affected livestock units and is detectable to a lesser degree in beetles and mites. Persistent carriage appears to be common and there is some field and experimental evidence to support arthropod-mediated transmission between poultry flocks, particularly carry-over from one flock to the next. Campylobacter may readily be isolated from arthropods in contact with affected poultry flocks, although carriage is short-lived. There appears to be a role for flies, at least, in the breaching of biosecurity around Campylobacter-negative flocks. The carriage of other zoonotic bacteria by arthropods has been documented, but the duration and significance of such associations remain uncertain in the context of livestock production.

Snow LC, Davies RH, Christiansen KH, Carrique-Mas JJ, Cook AJC, Evans SJ. 2010. Investigation of risk factors for Salmonella on commercial egg-laying farms in Great Britain, 2004-2005. Vet Rec, 166 (19), pp. 579-586. | Show Abstract | Read more

In 2004/05, all European Union member states were required to carry out standardised prevalence surveys to establish the baseline prevalence of Salmonella in commercial laying flocks. As part of the survey in Great Britain, additional data were collected from 380 of the enrolled laying hen holdings to investigate risk factors for Salmonella at farm level. Stratified, simple random sampling was used to select holdings from which dust and boot swab samples were collected and tested for Salmonella using a modification of ISO 6579:2002. Using a multivariable logistic model weighted to account for the survey design, several factors significantly associated with Salmonella and Salmonella Enteritidis status were identified. Larger holdings (>or=30,000 birds) were found to be at higher risk of Salmonella (odds ratio [OR] 4.79, P=0.025), while vaccination (OR 0.28, P=0.013), providing foot dips with brushes (OR 0.27, P=0.042), washing and disinfecting the house at depopulation (OR 0.19, P=0.003), having a clean car park away from house (OR 0.14, P=0.001), using an independent (OR 0.19, P=0.007) or other non-company (OR 0.40, P=0.049) source of feed, being over 1 km from the nearest neighbouring farm (OR 0.45, P=0.021) and the presence of cats and dogs on the farm (OR 0.26, P=0.002) or on contiguous farms (OR 0.44, P=0.030) reduced the risk of any Salmonella serovars being present. Factors found to be associated specifically with an increased risk of S Enteritidis infection included holding size (OR 14.88, P=0.001) and frequent sightings of rats (OR 8.17, P<0.001) or mice (OR 5.78, P=0.006). Non-caged systems (OR 0.14, P=0.002), vaccination (OR 0.08, P=0.001), the use of a non-company feed source (OR 0.11, P=0.003), running the site as all-in/all-out (OR 0.06, P<0.001) and the presence of cats and dogs on the farm (OR 0.14, P=0.002) were associated with a reduced risk.

Atterbury RJ, Davies RH, Carrique-Mas JJ, Morris V, Harrison D, Tucker V, Allen VM. 2010. Effect of delivery method on the efficacy of Salmonella vaccination in chickens. Vet Rec, 167 (5), pp. 161-164. | Show Abstract | Read more

To investigate whether the efficacy of live vaccines is influenced by the mode of vaccine delivery, a widely-used UK live commercial Salmonella Enteritidis vaccine was delivered to pullet chicks either by spray, in drinking water, or in combination with a bivalent vaccine containing inactivated Salmonella Enteritidis and Salmonella Typhimurium. The birds were subsequently challenged with 10(2) or 10(8) colony-forming units (cfu) of Salmonella Enteritidis through drinking water at either six or 20 weeks of age. Ten days after the challenge, the birds were euthanased and their caecal contents cultured for Salmonella. All of the vaccinated groups contained fewer Salmonella Enteritidis-positive birds than the unvaccinated groups. The 'spray-vaccinated' group contained significantly fewer Salmonella Enteritidis-positive birds than the 'water-vaccinated' group after challenge with 10(8) cfu at 20 weeks. However, there was little or no difference at the other challenge time points between the groups that received vaccine through different modes of delivery.

Featherstone CA, Reichel R, Snow LC, Davies RH, Christiansen KH, Carrique-Mas JJ, Evans SJ. 2010. Investigation of risk factors for Salmonella on fattening-turkey farms. Epidemiol Infect, 138 (10), pp. 1427-1438. | Show Abstract | Read more

A cross-sectional study into risk factors for Salmonella was undertaken using data gathered from 252 fattening turkey flocks in the UK. The data was derived from the EU baseline survey conducted during 2006 and 2007, in addition to a voluntary questionnaire. Multivariate logistic regression models identified significant risk factors for Salmonella spp. and Salmonella Typhimurium. A decreased risk of Salmonella spp. infection was associated with a history of intestinal illness in the sampled flock (OR 0.17), the use of wood shavings as litter (OR 0.21), use of disinfectant in the cleaning process (OR 0.25), incineration of dead birds on farm (OR 0.29), seasonal production (OR 0.31), farm staff also working with cattle (OR 0.31), and the presence of pigs on neighbouring farms (OR 0.38). The risk of isolating Salmonella spp. varied according to the company from which the poults were sourced. A reduced risk of S. Typhimurium infection was associated with the use of wax blocks to control rodents (OR 0.09), using mains water (OR 0.19) and having a Salmonella test programme (OR 0.23). An increased risk of S. Typhimurium infection was associated with storage of items around the turkey house (OR 5.20), evidence of mice (OR 4.71) and a soil surface surrounding the turkey house (OR 2.70). This study therefore identifies a number of important practical measures which can be implemented by farmers and veterinarians within the turkey industry to assist in the control of salmonellosis at the farm level.

Arnold ME, Papadopoulou C, Davies RH, Carrique-Mas JJ, Evans SJ, Hoinville LJ. 2010. Estimation of Salmonella prevalence in UK egg-laying holdings. Prev Vet Med, 94 (3-4), pp. 306-309. | Show Abstract | Read more

As part of an EU-wide programme to reduce the prevalence of Salmonella in commercial egg-laying holdings, the EU has set for the UK an annual target of 10% reduction in the prevalence of Salmonella Enteritidis and Salmonella Typhimurium in commercial egg-laying holdings. To assist in demonstrating such a reduction, it is very important to obtain an accurate as possible baseline prevalence for Salmonella. The objective of this study was to provide a baseline estimate of the Salmonella prevalence in egg-laying holdings in the UK. Data from an EU baseline survey for Salmonella in UK commercial egg-laying flocks were therefore analysed using Bayesian methods, taking into account the sampling of only 1 flock per holding and estimates of the test sensitivity of the methods used in the EU baseline survey. In addition, in the UK the majority of the eggs come from farms which have participated in voluntary monitoring programmes for Salmonella since the early 1990s, and this data was also used, along with a prior estimate of the test sensitivity of voluntary surveillance. Results indicated that a true prevalence 14% for Salmonella Enteriditis and Typhimurium, and 18% for all serovars, both of these estimates being higher than has previously been reported from the EU baseline survey data. It is also shown that the sensitivity of voluntary surveillance is low, and it will therefore be important to compare results from "official" and "non-official" samples to check that the sampling performed in the National Control Plan is as sensitive as expected.

Mueller-Doblies D, Carrique-Mas JJ, Sayers AR, Davies RH. 2010. A comparison of the efficacy of different disinfection methods in eliminating Salmonella contamination from turkey houses. J Appl Microbiol, 109 (2), pp. 471-479. | Show Abstract | Read more

AIMS: This study aimed to compare the efficacy of different disinfection methods in eliminating Salmonella contamination from turkey houses. METHODS AND RESULTS: Fifty depopulated turkey houses which had all housed Salmonella-positive flocks were visited after cleaning and disinfection. A minimum of 45 swab samples from different surfaces were taken per house and analysed for the presence of Salmonella. The sampled surfaces included intact floor surfaces, floor cracks, walls, feeders, drinkers, anteroom, nestboxes and miscellaneous items. Houses were grouped according to the disinfectant which had been used and the efficacy of the different groups of disinfectants was compared. Sixty-eight % of houses tested positive for Salmonella after C&D. Out of 4440 samples, 207 tested positive for Salmonella, giving an overall sample prevalence of 4.7%. There was no significant difference in the level of residual contamination between breeding, rearing and finishing houses. Products containing a mixture of formaldehyde, glutaraldehyde and quaternary ammonium compounds (QAC) performed significantly better than products containing hydrogen peroxide and peracetic acid. Cleaning and disinfection was least effective in nestboxes and anterooms. CONCLUSIONS: Thorough cleaning and the choice of a suitable disinfectant are crucial if Salmonella contamination of turkey houses is to be eliminated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that disinfectants containing a mixture of formaldehyde, glutaraldehyde and QAC perform significantly better under field conditions than oxidising products and should therefore be the first choice for disinfection of turkey premises where Salmonella is present.

Arnold ME, Mueller-Doblies D, Carrique-Mas JJ, Davies RH. 2009. The estimation of pooled-sample sensitivity for detection of Salmonella in turkey flocks. J Appl Microbiol, 107 (3), pp. 936-943. | Show Abstract | Read more

AIMS: To investigate the effectiveness of pooled sampling methods for detection of Salmonella in turkey flocks. METHODS AND RESULTS: Individual turkey droppings were taken from 43 flocks, with half the dropping tested for Salmonella as an individual sample and the other half included in a pool of five. A pair of boot swabs and a dust sample were also taken from each flock. The results were analysed using Bayesian methods in the absence of a gold standard. This showed a dilution effect of mixing true-positive with negative samples, but even with this the pooled faecal samples were found to be a highly efficient method of testing compared with individual faecal samples. The more samples included in the pool, the more sensitive the pooled sampling method was predicted to be. The sensitivity of dust sampling was much more sensitive than faecal sampling at low prevalence. CONCLUSIONS: Pooled faecal sampling is an efficient method of Salmonella detection in turkey flocks. The additional testing of a dust sample greatly increased the effectiveness of sampling, especially at low prevalence. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to relate the sensitivity of the sampling methods to the within-flock prevalence.

Carrique-Mas JJ, Marin C, Breslin M, McLaren I, Davies R. 2009. A comparison of the efficacy of cleaning and disinfection methods in eliminating Salmonella spp. from commercial egg laying houses. Avian Pathol, 38 (5), pp. 419-424. | Show Abstract | Read more

Effective terminal cleaning and disinfection (C&D) is regarded as a necessary step for the elimination of Salmonella spp. from laying houses. A total of 60 commercial laying houses that had housed laying flocks infected with Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium that were representative of all production systems (cage, barn, free-range) were intensively sampled immediately after C&D as well as in the follow-on flock. The procedures investigated were: (1) a compound disinfectant consisting of a mixture of formaldehyde, glutaraldehyde and quarternary ammonium applied at the recommended concentration; (2) a 10% (vol/vol) dilution of the standard 37% commercial formalin, applied by a contractor; and (3) other disinfection procedures selected and applied by the farmer. The recovery of Salmonella in the cleaned and disinfected houses was variable, with samples from floor and dropping boards/belts (cage houses) and scratching areas (non-cage houses) being the most likely to remain contaminated. In cage houses, the use of the 10% formalin dilution led to a statistically greater reduction in the sample prevalence than using any of the other C&D methods. A negative post-C&D result predicted clearance of Salmonella in 52% of cases, although the isolation of Salmonella from the houses immediately after C&D was not a perfect predictor of carry-over of infection.

Mueller-Doblies D, Sayers AR, Carrique-Mas JJ, Davies RH. 2009. Comparison of sampling methods to detect Salmonella infection of turkey flocks. J Appl Microbiol, 107 (2), pp. 635-645. | Show Abstract | Read more

AIMS: To compare the efficiency of various sampling methods for detection of Salmonella in turkey flocks. METHODS AND RESULTS: In a field study that compared various sampling methods one pair of boot swabs taken from the whole turkey house provided suitably sensitive results for fattening and rearing flocks and was no less sensitive than two pairs, each from half the house, tested as a pooled sample. The sensitivity was further enhanced by adding a dust sample. The dust sample appeared to be particularly useful in flocks with a low prevalence, especially in breeding flocks, and was more sensitive than a method which used five pairs of boot swabs per flock. Combined incubation of a boot swab and a dust sample showed no interference between the two sample types and a maximum sensitivity of detection. Litter samples and commercial sponge drag swabs provided a lower level of detection. CONCLUSIONS: A single pair of boot swabs taken from the whole house is recommended for routine sampling of commercial rearing or fattening flocks. An additional dust sample could be added to increase detection in flocks with a low prevalence or in breeding flocks, but adding an additional pair of boot swabs would not increase detection compared with a single pair. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that significant efficiencies can be made in sampling programmes for detection of Salmonella in turkey flocks without detracting from the sensitivity. Similar studies are recommended for other poultry sectors, particularly in chicken breeding flocks.

Carrique-Mas JJ, Barnes S, McLaren I, Davies R. 2009. Comparison of three plating media for the isolation of Salmonella from poultry environmental samples in Great Britain using ISO 6579:2002 (Annex D). J Appl Microbiol, 107 (6), pp. 1976-1983. | Show Abstract | Read more

AIMS: To evaluate the performance of three Salmonella plating media (Rambach, Xylose Lysine Deoxycholate agar and modified Brilliant Green Agar plus Novobiocin) as part of the ISO 6579: 2002 (Annex D) on poultry environmental samples. METHODS AND RESULTS: The samples analysed were those for the European Union Salmonella baseline surveys of laying (N = 3087), broiler (N = 1550), turkey fattening (N = 1540) and turkey breeding (N = 580) flocks for Great Britain. Results were considered separately for Rambach (including and excluding pale orange colonies) and for growth on selective media [Modified semi-solid Rappaport Vassiliadis (MSRV)] after 24 and 48 h of incubation. Overall, Rambach was the most sensitive medium, provided that pale orange colonies were checked. In all cases, an increase in the sensitivity of detection was obtained by plating growth on MSRV after 48 h of incubation. In broilers and laying flocks, the specificity significantly improved when Rambach only was used. CONCLUSION: The use of Rambach results in considerable savings compared with the two-plate method prescribed by ISO 6579:2002 (Annex D) without compromising sensitivity. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella isolation protocols should be reviewed in terms of their efficiency and cost.

Wales AD, McLaren IM, Bedford S, Carrique-Mas JJ, Cook AJC, Davies RH. 2009. Longitudinal survey of the occurrence of Salmonella in pigs and the environment of nucleus breeder and multiplier pig herds in England. Vet Rec, 165 (22), pp. 648-657. | Show Abstract | Read more

Eight pig breeding units previously associated with Salmonella Typhimurium were visited during a period of up to seven years. Samples from voided faeces, surfaces, fomites and wildlife were cultured. Certain serovars (Derby, Stanley, Give, Bredeney, Mbandaka and Manhattan) were isolated repeatedly on certain units, while others (Agona, Ajiobo, Heidelberg, Meleagridis, Muenchen, Montevideo, Rissen and Senftenberg) were detected only once or intermittently. Serovars Kedougou, Newport and Typhimurium were isolated consistently on some units but only intermittently on others. There was an association between the Salmonella serovar in pens and in the immediate environment of the pens. Pens holding breeding stock destined for production herds were frequently positive for Salmonella. Herds under common ownership showed similar serovar combinations. Serovars from wildlife were typical of the associated premises. Cleaning and disinfection was frequently ineffective. On one unit, a low level of Salmonella was attributed to a small herd size, good cleaning and disinfection, and good rodent control. Breeding herds are therefore susceptible to endemic infections with multiple Salmonella serovars, and cleaning, disinfection and vector control may be inadequate in many cases. The prevalence of S Typhimurium was greater in youngstock, which may have important implications for public health.

Papadopoulou C, Carrique-Mas JJ, Davies RH, Sayers AR. 2009. Retrospective analysis of Salmonella isolates recovered from animal feed in Great Britain. Vet Rec, 165 (23), pp. 681-688. | Show Abstract

To examine feed contamination rates with Salmonella, the diversity of serovars and the antimicrobial resistance of isolates from animal feedingstuffs in Great Britain, and to compare Salmonella strains found in animal feed and in livestock, data collected under voluntary and statutory Salmonella surveillance during the period 1987 to 2006 were analysed retrospectively. The feed contamination rate decreased from 3.8 per cent in 1993 to 1.1 per cent in 2006. A total of 263 Salmonella serovars were recovered: S Mbandaka (11.2 per cent), S Tennessee (10.4 per cent), S Senftenberg (8.4 per cent), S Agona (6.4 per cent), S Montevideo (6.4 per cent) and S Ohio (3.1 per cent) were the most prevalent. S Typhimurium was recovered at a proportion of 1.6 per cent from raw ingredients and 2.4 per cent from finished feed, while S Enteritidis was recovered at a proportion of 0.5 per cent from raw ingredients and 0.6 per cent from finished feed; 14.1 per cent of the isolates were resistant to at least one antimicrobial, and 1.9 per cent were multiresistant. There was no evidence of a statistical association (P<0.05) between the top 10 serovars recovered from feed and from livestock.

Papadopoulou C, Davies RH, Carrique-Mas JJ, Evans SJ. 2009. Salmonella serovars and their antimicrobial resistance in British turkey flocks in 1995 to 2006. Avian Pathol, 38 (5), pp. 349-357. | Show Abstract | Read more

Serovar and antimicrobial resistance data from the scanning surveillance of British turkey flocks for Salmonella between 1995 and 2006 were analysed and compared with prevalence data from other livestock and animal feed. A total of 2753 incidents of 57 different serovars were reported. The five most prevalent serovars were Salmonella Typhimurium (20.8%), Salmonella Newport (14.7%), Salmonella Derby (10.6%), Salmonella Indiana (8.3%) and Salmonella Agona (6.4%). S. Typhimurium reports peaked in the mid- to late 1990s; this occurred in parallel with the S. Typhimurium DT104 epidemic in other livestock species. S. Enteritidis reports peaked in mid- to late 1990s, followed by a considerable decrease after 2000, which was also noted in flocks of domestic fowl. S. Newport, Salmonella Montevideo, Salmonella Senftenberg and Salmonella Binza occurred in marked clusters, indicating that they were introduced into one or more flocks at a certain time (i.e. via contaminated feed or infected 1-day-old chicks). A proportion of 43.1% of the reported Salmonella isolates were resistant to at least one antimicrobial, while 17.7% were multi-resistant. No isolates were resistant to ciprofloxacin or to the third-generation cephalosporins ceftazidime and cefotaxime. Resistance to ampicillin, chloramphenicol, streptomycin, sulphonamide compounds and tetracycline was common, and it was mainly a characteristic of S. Typhimurium DT104 compared with S. Typhimurium non-DT104 and non-S. Typhimurium isolates (P<0.001). Resistance to nalidixic acid decreased from 16.9% in 1995 to 11.8% in 2006. Nalidixic acid resistance was most frequently found in Salonella Hadar (71.4%), S. Typhimurium DT104 (30.0%), S. Newport (17.9%) and S. Typhimurium non-DT104 (11.1%).

Atterbury RJ, Carrique-Mas JJ, Davies RH, Allen VM. 2009. Salmonella colonisation of laying hens following vaccination with killed and live attenuated commercial Salmonella vaccines. Vet Rec, 165 (17), pp. 493-496. | Show Abstract | Read more

The aim of this study was to determine the efficacy of a killed Salmonella vaccine and three live vaccines in preventing caecal colonisation of Hy-line Brown pullets by Salmonella Enteritidis PT 4. The lowest number of Salmonella-positive birds following the largest challenge (10(8) cfu) was recorded for live vaccine 1. However, birds treated with the killed vaccine had a significantly lower number of salmonellae in their caeca compared with both the control group and the other vaccine groups (P<0.05).

Carrique-Mas JJ, Breslin M, Snow L, McLaren I, Sayers AR, Davies RH. 2009. Persistence and clearance of different Salmonella serovars in buildings housing laying hens. Epidemiol Infect, 137 (6), pp. 837-846. | Show Abstract | Read more

We investigated factors associated with persistence of different Salmonella serovars in buildings housing laying hens in Great Britain using survival analysis. A total of 264 incidents of Salmonella detection occurring between July 1998 and August 2007 in 152 houses were recorded. For incidents involving Salmonella Enteritidis (SE), both the rodent score of the house and the type of house were positively associated with persistence. For non-SE serovars, only the type of house was associated with persistence. Persistence of SE in the houses was longest (>15 months) in step-cage and cage-scraper houses when high levels of rodents were present, and lowest in non-cage and cage-belt houses. We estimated that 42% (95% CI 23.3-63.1) of SE incidents may be cleared during the lay period, and this was related to elimination of rodents from the houses. From January 2009, EU legislation will ban the sale of fresh eggs from SE-positive and S. Typhimurium-positive flocks over their remaining lifespan. If infection is eliminated from such flocks, they would cease to represent a public health risk.

Carrique-Mas JJ, Davies RH. 2008. Salmonella Enteritidis in commercial layer flocks in Europe: legislative background, on-farm sampling and main challenges Revista Brasileira de Ciência Avícola, 10 (1), pp. 1-9. | Read more

Carrique-Mas JJ, Medley GF, Green LE. 2008. Risks for bovine tuberculosis in British cattle farms restocked after the foot and mouth disease epidemic of 2001. Prev Vet Med, 84 (1-2), pp. 85-93. | Show Abstract | Read more

The foot and mouth disease (FMD) epidemic of 2001 was used to investigate herd breakdown (HBD) with bovine tuberculosis (bTB) in totally restocked herds of cattle. By August 2004, 2941 restocked cattle herds, with cattle movements from before and after 2001, had been tested for bTB for the first time since restocking. A total of 6% (177) of these herds broke down at the first bTB test. A binomial logistic regression model with HBD (at least one reactor bovine) at the first test after restocking as the outcome was used to investigate risks associated with HBD. The final model contained three risk factors. There was an increased risk for HBD in restocked herds with every log increase in herd size with an OR=1.38 (CI 1.16-1.64) to a maximum OR of 10.75. When there was a history of bTB on the restocked farm before 2001 the OR, with CI not including unity, were 5.92, 4.63, 3.8 and 2.9 for last HBD in 2000, 1999, 1998 and 1997, respectively, indicating a persistence in increased risk for restocked herds from farms with a history of HBD in the previous herd before restocking, i.e. a different population of cattle. Finally, for every log increase in the number of cattle purchased from herds with a greater than biennial frequency of testing for bTB in the previous 8 years (i.e. perceived high risk herds for bTB) there was an OR=1.35 (95% CI 1.22-1.49). The maximum OR was 9.27. These results indicate that both introduction of bTB through the purchase of cattle from farms with a high perceived risk of bTB infection and persistence of bTB on the restocked farm, (not the farm's original herd), were associated with an increased risk of HBD in the newly formed herds after restocking.

Carrique-Mas JJ, Davies RH. 2008. Bacteriological detection of Salmonella enteritidis in eggs: a review. Rev Sci Tech, 27 (3), pp. 657-664. | Show Abstract | Read more

The detection of Salmonella enterica serovar Enteritidis (SE) in eggs is hampered by a typically low prevalence of contaminated eggs, the low number of SE organisms in such eggs, and the presence of inhibitory substances in the egg albumin. For these reasons, the analysis of large pools of eggs is normally necessary, which presents logistic and microbiological challenges associated with a low number of target organisms from a large volume of sample matrix. In some studies using artificially inoculated eggs the standard procedure for Salmonella culture consisting of pre-enrichment, followed by selective enrichment and plating has been replaced by incubation of the egg pools at 25 degrees C to 37 degrees C followed by direct plating. However, in most cases using pools of naturally contaminated eggs, it may be necessary to enhance the traditional three-step method by addition of antibiotics or iron supplements.

Snow LC, Davies RH, Christiansen KH, Carrique-Mas JJ, Cook AJC, Teale CJ, Evans SJ. 2008. Survey of the prevalence of Salmonella on commercial broiler farms in the United Kingdom, 2005/06. Vet Rec, 163 (22), pp. 649-654. | Show Abstract | Read more

Between October 2005 and September 2006, all European Union member states were required to carry out standardised surveys of the prevalence of Salmonella in broiler flock holdings to establish baseline data from which to derive national targets for disease reduction. In the uk 382 holdings were sampled, 41 of which were positive for Salmonella, giving an estimated weighted prevalence of 10.7 per cent (95 per cent confidence interval [ci] 8.1 to 13.1 per cent). The serotype most frequently isolated was Salmonella Ohio, with a weighted prevalence of 2.2 per cent (95 per cent ci 1.2 to 3.7 per cent), followed by Salmonella Kedougou at 1.7 per cent (95 per cent ci 0.9 to 3.2 per cent). There were no isolates of Salmonella Enteritidis and only a single isolation of Salmonella Typhimurium (0.2 per cent, 95 per cent ci 0.0 to 1.6 per cent). Of the three other serotypes given top priority by the eu owing to their public health significance, Salmonella Virchow was isolated from one holding, but Salmonella Hadar and Salmonella Infantis were not detected on any of the holdings.

Carrique-Mas JJ, Davies RH. 2008. Sampling and bacteriological detection of Salmonella in poultry and poultry premises: a review. Rev Sci Tech, 27 (3), pp. 665-677. | Show Abstract | Read more

The detection of Salmonella in primary poultry production is an issue of great concern in the European Union (EU), since control of this zoonotic disease is in part based on the reduction of the prevalence at the farm level. Success of detection is likely to be highly dependent on the choice of an adequate sampling procedure combined with a sensitive culture method. In poultry farms 'naturally pooled' faeces/litter and dust are the matrices of choice. In floor systems boot swabs are the preferred method for the collection of faeces. A wide range of culture methods is available, but ISO 6579:2002 (Annex D) is currently the standard for poultry environmental samples in the EU. In this review, the authors discuss in detail the range of sampling and culture methodologies for Salmonella in poultry farms. The review also covers sampling and testing of poultry hatcheries, live birds and poultry carcasses.

Carrique-Mas JJ, Papadopoulou C, Evans SJ, Wales A, Teale CJ, Davies RH. 2008. Trends in phage types and antimicrobial resistance of Salmonella enterica serovar Enteritidis isolated from animals in Great Britain from 1990 to 2005. Vet Rec, 162 (17), pp. 541-546. | Show Abstract | Read more

Surveillance data for Salmonella enterica serovar Enteritidis incidents and isolations from food animals in Great Britain from 1990 to 2005 were analysed to detect any trends and provide the basis for a comparison between phage types (pt) and antimicrobial sensitivity patterns in human beings and animals. During 2001 to 2005 there was a decrease in incidents involving most species except ducks. Only the numbers of incidents involving pts 6, 6a, 9b and 14b (in ducks) and pts 6a and 13a (in mammals) increased significantly during this period, whereas there were 93 per cent fewer incidents involving pt 4 than in 1990 to 2000. After adjustment for pt, the isolates from ducks were more resistant to nalidixic acid, tetracyclines and sulfonamides, and were more likely to be multiresistant than isolates from chickens. Isolates from turkeys tended to be more resistant to sulfonamides than isolates from chickens. pts 1, 5a, 6, 6a and 35 had the highest level of resistance after adjusting for species. During 2001 to 2005 there was an increase in resistance among pts 1, 6 and 7, in most cases involving nalidixic acid.

Carrique-Mas JJ, Breslin M, Snow L, Arnold ME, Wales A, McLaren I, Davies RH. 2008. Observations related to the Salmonella EU layer baseline survey in the United Kingdom: follow-up of positive flocks and sensitivity issues. Epidemiol Infect, 136 (11), pp. 1537-1546. | Show Abstract | Read more

A follow-on study was carried out on 23 holdings identified as Salmonella positive in the 2004/2005 European Union (EU) baseline survey of Salmonella in laying hens. Eleven of 13 cage and 4/7 floor houses remained positive for Salmonella when the new flock was tested, and from 10/13 cage and 3/7 floor houses a Salmonella of the same serovar/phage type as found in the EU survey was isolated. There was a high correlation between the level of contamination in the houses at the time of the EU survey and in the follow-on flock. On seven occasions the house identified as positive in the EU survey was sampled after cleaning and disinfection but before a new flock was placed, and in all of them Salmonella could be isolated from the houses. The observed number of infected houses in infected holdings suggests that the holding-level prevalence in the United Kingdom would be about 21% higher than the results obtained in the EU survey.

Carrique-Mas JJ, Breslin M, Sayers AR, McLaren I, Arnold M, Davies R. 2008. Comparison of environmental sampling methods for detecting Salmonella in commercial laying flocks in the UK. Lett Appl Microbiol, 47 (6), pp. 514-519. | Show Abstract | Read more

AIMS: To investigate the performance of the Salmonella National Control Programme (NCP) sampling/testing methods in laying flocks of domestic fowl. METHODS AND RESULTS: Eighty-five visits were made to 69 flocks representative of the main production systems (cage, barn and free-range) infected with Salmonella. In each visit, three methodologies were compared: (i) the European Union (EU) baseline survey method (five faeces and two dust samples); (ii) an in-house (Veterinary Laboratories Agency, VLA) 'wet' method that involved collecting 10 dust and 10 faeces samples into jars with buffered peptone water; and (iii) a method involving two samples of pooled faeces and one of dust (cultured as one sample of each type), which has been adopted for the NCP for laying flocks across the EU. CONCLUSIONS: The 'wet' method was the most sensitive, and the NCP the least, although individual NCP samples were the most sensitive ones. SIGNIFICANCE AND IMPACT OF THE STUDY: The apparent lower sensitivity of the NCP method may be compensated by repeated sampling of flocks (twice during rear and several times during lay). Sampling using VLA methodology should be advocated for farms aiming to disclose low-level Salmonella before restrictions on the sale of eggs from Salmonella Enteritidis or Salmonella Typhimurium-infected flocks are in place.

Snow LC, Davies RH, Christiansen KH, Carrique-Mas JJ, Wales AD, O'Connor JL, Cook AJC, Evans SJ. 2007. Survey of the prevalence of Salmonella species on commercial laying farms in the United Kingdom. Vet Rec, 161 (14), pp. 471-476. | Show Abstract | Read more

A survey of salmonella infection on 454 commercial layer flock holdings in the uk was carried out between October 2004 and September 2005. Fifty-four (11.7 per cent, 95 per cent confidence interval 9.3 to 14.0 per cent) were salmonella positive. The most common serovar identified was Salmonella Enteritidis at a prevalence of 5.8 per cent, and 70 per cent of these isolates were phage types 4, 6, 7 and 35. Salmonella Typhimurium was the second most prevalent serovar, found in 1.8 per cent of the farms. Of the three other serovars given top priority by the eu because of their public health significance, Salmonella Virchow and Salmonella Infantis were each isolated from one holding, but Salmonella Hadar was not isolated from any of the holdings. Analysis of antimicrobial resistance patterns revealed that over 76 per cent of the isolates were sensitive to all of the 16 drugs tested, and all the isolates were sensitive to ciprofloxacin, gentamicin, ceftazidime, apramycin, amikacin, amoxicillin/clavulanic acid, neomycin and cefotaxime.

Carrique-Mas JJ, Bedford S, Davies RH. 2007. Organic acid and formaldehyde treatment of animal feeds to control Salmonella: efficacy and masking during culture. J Appl Microbiol, 103 (1), pp. 88-96. | Show Abstract | Read more

AIMS: To investigate whether treatment of animals feeds with organic acids/formaldehyde may mask the presence of Salmonella, when assessed by standard cultural methods. METHODS AND RESULTS: Four commercial treatments were applied at the manufacturers' recommended rates on feeds artificially inoculated with Salmonella. The recovery of Salmonella from these treated feeds was assessed after specific antagonists were added to the treatments during culture. A control group of treated feed received no antagonist. Masking of Salmonella was demonstrated when the addition of antagonists resulted in recovery of Salmonella from the treated feed, compared with a negative recovery when no antagonists were added. There were large variations in the efficacy of treatments, and masking was demonstrated with all four tested treatments. One formaldehyde-based product showed greater efficacy and less masking. Masking was greater when high levels of Salmonella were present in the feed. CONCLUSIONS: Some organic acid or formaldehyde-based feed treatments may mask the presence of Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: Feeds may be deemed safe despite being contaminated with Salmonella. The use of antagonists during culture may help assess the level of Salmonella contamination when organic acid or formaldehyde treatments have been applied to feed ingredients.

Carrique-Mas J, Andersson Y, Hjertqvist M, Svensson A, Torner A, Giesecke J. 2005. Risk factors for domestic sporadic campylobacteriosis among young children in Sweden. Scand J Infect Dis, 37 (2), pp. 101-110. | Show Abstract | Read more

A case-control study was conducted in Sweden to study risk factors for domestically acquired Campylobacter jejuni/coli infections among children aged less than 6 y. A total of 126 cases, reported to the national surveillance system were recruited over 1 y. Controls, selected from the population register, were matched to the cases by age, gender, place of residence and time of infection of the case. Information was gathered by posted questionnaires. Two separate conditional regression models were developed including and excluding 'protective' factors. Two of the factors significantly associated with Campylobacter infection were water-related: having a well in the household (OR=2.6) and drinking water from a lake/river (OR=7.4; 6.0). Other exposures associated with increased risk were: having a dog (OR=8.4; 3.8) and eating grilled meat (OR=5.5; 2.1). Drinking unpasteurized milk was borderline significant in 1 model (OR=3.7). Eating sausage was protective (OR=0.05). Eating chicken was not a significant risk. Exposures such as eating grilled meat and drinking water from a lake or a river were more common in the warm months, a factor that may partly explain the observed seasonality. The authors suggest that differences between risk factors across studies may reflect geographical and age-specific differences in the sources of infection.

Carrique-Mas J, Andersson Y, Petersén B, Hedlund KO, Sjögren N, Giesecke J. 2003. A norwalk-like virus waterborne community outbreak in a Swedish village during peak holiday season. Epidemiol Infect, 131 (1), pp. 737-744. | Show Abstract | Read more

An outbreak of gastroenteritis due to Norwalk-like virus (NLV) affecting approximately 500 people occurred in a Swedish ski resort during February-March 2002. Epidemiological investigations were performed on cohorts of schoolchildren, permanent residents and skiers visiting the area. Attack rates were respectively 39.7, 29.9 and 38.5%. Drinking un-boiled water originating from one of the three communal water systems was a significant risk factor for all groups. For schoolchildren, the risk of illness increased with increasing amount of water consumed. Nine of 12 stool samples of patients analysed tested positive for NLV. The water tested negative for indicator bacteria and results of NLV tests were inconclusive. In the absence of microbiological findings, the environmental authorities were reluctant to act based on the epidemiological analysis alone, and intervention was delayed until mid-April, following the discovery of a crack in a sewage pipe 10 m from the well.

Carrique-Mas JJ, Hökeberg I, Andersson Y, Arneborn M, Tham W, Danielsson-Tham ML, Osterman B, Leffler M, Steen M, Eriksson E et al. 2003. Febrile gastroenteritis after eating on-farm manufactured fresh cheese--an outbreak of listeriosis? Epidemiol Infect, 130 (1), pp. 79-86. | Show Abstract | Read more

An outbreak of febrile gastroenteritis affected consumers of on-farm manufactured dairy products from a summer farm in Sweden. Symptoms included diarrhoea, fever, stomach cramps and vomiting in 88, 60, 54 and 21% of cases identified. The median incubation period was 31 h. A cohort study with 33 consumers showed an attack rate of 52% and an association between the total amount of product eaten and illness (P=0.07). Twenty-seven of 32 (84%) stool samples cultured for Listeria monocytogenes tested positive, although there was no association between clinical disease and the isolation of L. monocytogenes. In addition, gene sequences for VTEC and ETEC were detected in 6 and 1 subjects, respectively. Bacteriological analysis of cheese samples revealed heavy contamination with L. monocytogenes and coagulase positive staphylococci in all of them and gene markers for VTEC in one of them. Molecular profiles for L. monocytogenes isolated from dairy products, stool samples and an abscess from 1 patient who developed septic arthritis were identical. Results of both microbiological and epidemiological analyses point to L. monocytogenes as the most likely cause of this outbreak. The finding of markers for VTEC in some humans and cheese samples means that a mixed aetiology at least in some cases cannot be conclusively ruled out.

Carrique-Mas J, Iihoshi N, Widdowson MA, Roca Y, Morales G, Quiroga J, Cejas F, Caihuara M, Ibarra R, Edelsten M. 2001. An epidemiological study of Taenia solium cysticercosis in a rural population in the Bolivian Chaco. Acta Trop, 80 (3), pp. 229-235. | Show Abstract | Read more

A survey of 100 rural households in a village in the Chaco region of Bolivia revealed a serious problem of Taenia solium cysticercosis, with a seroprevalence of 99/447 (22%) in humans and 102/273 (37%) in pigs. Risk factors for humans were being in older age groups, absence of sanitary facilities, poor formal education and inability to recognise infected pork. Significant risk indicators were a history of seizures and the reported elimination of worms in the faeces. Risk factors for pigs were being in older age groups and absence of sanitary facilities in the owner's house. The proportion of households with evidence of human cysticercosis was similar for those who owned pigs (48%) and those that did not (55%). This unexpected finding was attributed to the high overall prevalence of cysticercosis in pigs and the probability that everyone, regardless of pig-ownership, had ample opportunity to become infected in such communities. The main recommendation for reducing the prevalence of human cysticercosis was to provide more effective education campaigns, aimed at preventing both T. solium infection and cysticercosis.

Carrique Mas JJ, Widdowson MA, Cuéllar AM, Ribera H, Walker AR. 2000. Risk of babesiosis and anaplasmosis in different ecological zones of Santa Cruz Department, Bolivia. Vet Parasitol, 93 (1), pp. 29-38. | Show Abstract | Read more

A cross-sectional study was done of seroprevalence of Babesia bigemina, B.bovis, and Anaplasma marginale in cattle from eastern Bolivia, to characterise the risk of tick-borne disease in three ecological zones. Nineteen farms were sampled in the subtropical humid zone, 13 in the dry subtropical zone and nine in the lower western valleys of the Andean massif. The enzyme-linked immunosorbent assay was used. All three pathogens were widespread. For B. bovis, seroprevalences were high (75-78%) in the two subtropical zones which thus had low risk of disease from this infection; but the western valleys were endemically unstable with higher risk. For B. bigemina, seroprevalences were lower (24-57%) in the two subtropical zones and thus these areas were endemically unstable for disease from this infection. However, the seroprevalence of B. bigemina in the western valleys was too low (13%) for risk of disease in susceptible cattle to be considered high. For A. marginale, the seroprevalences in the two subtropical zones were low (19-32%) and very low (6%) in the western valleys suggesting all these zones were endemically unstable for anaplasmosis. Data for individual farms were analysed for risk of both forms of babesiosis; this showed low risk of disease in the subtropical humid zone, higher risk in the dry subtropical zone and variable risk in the western valleys.

Nguyen NT, Nguyen HM, Nguyen CV, Nguyen TV, Nguyen MT, Thai HQ, Ho MH, Thwaites G, Ngo HT, Baker S, Carrique-Mas J. 2016. Use of Colistin and Other Critical Antimicrobials on Pig and Chicken Farms in Southern Vietnam and Its Association with Resistance in Commensal Escherichia coli Bacteria. Appl Environ Microbiol, 82 (13), pp. 3727-3735. | Show Abstract | Read more

UNLABELLED: Antimicrobial resistance (AMR) is a global health problem, and emerging semi-intensive farming systems in Southeast Asia are major contributors to the AMR burden. We accessed 12 pig and chicken farms at key stages of production in Tien Giang Province, Vietnam, to measure antimicrobial usage and to investigate the prevalence of AMR to five critical antimicrobials (β-lactams, third-generation cephalosporins, quinolones, aminoglycosides, and polymyxins) and their corresponding molecular mechanisms among 180 Escherichia coli isolates. Overall, 94.7 mg (interquartile range [IQR], 65.3 to 151.1) and 563.6 mg (IQR, 398.9 to 943.6) of antimicrobials was used to produce 1 kg (live weight) of chicken and pig, respectively. A median of 3 (out of 8) critical antimicrobials were used on pig farms. E. coli isolates exhibited a high prevalence of resistance to ampicillin (97.8% and 94.4% for chickens and pigs, respectively), ciprofloxacin (73.3% and 21.1%), gentamicin (42.2% and 35.6%), and colistin (22.2% and 24.4%). The prevalence of a recently discovered colistin resistance gene, mcr-1, was 19 to 22% and had strong agreement with phenotypic colistin resistance. We conducted plasmid conjugation experiments with 37 mcr-1 gene-positive E. coli isolates and successfully observed transfer of the gene in 54.0% of isolates through a plasmid of approximately 63 kb, consistent with one recently identified in China. We found no significant correlation between total use of antimicrobials at the farm level and AMR. These data provide additional insight into the role of mcr-1 in colistin resistance on farms and outline the dynamics of phenotypic and genotypic AMR in semi-intensive farming systems in Vietnam. IMPORTANCE: Our study provides accurate baseline information on levels of antimicrobial use, as well as on the dynamics of phenotypic and genotypic resistance for antimicrobials of critical importance among E. coli over the different stages of production in emerging pig and poultry production systems in Vietnam. E. coli isolates showed a high prevalence of resistance (>20%) to critically important antimicrobials, such as colistin, ciprofloxacin, and gentamicin. The underlying genetic mechanisms identified for colistin (the mcr-1 gene) and quinolone (gyrA gene mutations) are likely to play a major role in AMR to those compounds. Conjugation experiments led to the identification of a 63-kb plasmid, similar to one recently identified in China, as the potential carrier of the mcr-1 gene. These results should encourage greater restrictions of such antimicrobials in Southeast Asian farming systems.

Walther BA, Boëte C, Binot A, By Y, Cappelle J, Carrique-Mas J, Chou M, Furey N, Kim S, Lajaunie C et al. 2016. Biodiversity and health: Lessons and recommendations from an interdisciplinary conference to advise Southeast Asian research, society and policy. Infect Genet Evol, 40 pp. 29-46. | Show Abstract | Read more

Southeast Asia is an economic, biodiverse, cultural and disease hotspot. Due to rapid socio-economic and environmental changes, the role of biodiversity and ecosystems for human health ought to be examined and communicated to decision-makers and the public. We therefore summarized the lessons and recommendations from an interdisciplinary conference convened in Cambodia in 2014 to advise Southeast Asian societies on current research efforts, future research needs, and to provide suggestions for improved education, training and science-policy interactions. First, we reviewed several examples of the important role of ecosystems as 'sentinels' in the sense that potentially harmful developments for human health become first apparent in ecosystem components. Other ecosystem services which also benefit human well-being are briefly summarized. Second, we summarized the recommendations of the conference's roundtable discussions and added recent developments in the science-policy interface. The recommendations were organized along five themes: Ethical and legal considerations; implementation of the One Health approach; education, training, and capacity building; future research priorities; and potential science-policy interactions. While the role of biodiversity for human health needs further research, especially for zoonoses and emerging diseases, many direct and indirect benefits to human health are already apparent, but have yet to filter down to the science-policy interface in order to influence legislation and enforcement. Therefore, efforts to strengthen the interface in Southeast Asia should become a high priority in order to strengthen the health and resilience of Southeast Asian societies.

Van Cuong N, Nhung NT, Nghia NH, Mai Hoa NT, Trung NV, Thwaites G, Carrique-Mas J. 2016. Antimicrobial Consumption in Medicated Feeds in Vietnamese Pig and Poultry Production. Ecohealth, 13 (3), pp. 490-498. | Show Abstract | Read more

Antimicrobials are extensively used as growth promoters in animal feeds worldwide, but reliable estimates are lacking. We conducted an internet-based survey of commercial chicken and pig feed products officially approved for sale in Vietnam over the period March-June 2015. Information on the antimicrobial contents in feed products, alongside animal production data, was used to estimate in-feed antimicrobial consumption to produce one kilogram of live animal (chicken, pig), as well as to estimate country-wide antimicrobial consumption through animal feeds. A total of 1462 commercial feed formulations were examined. The survey-adjusted estimated antimicrobial contents were 25.7 and 62.3 mg/kg in chicken and pig feeds, respectively. Overall, it was estimated that 77.4 mg [95% CI 48.1-106.8] and 286.6 mg [95% CI 191.6-418.3] of in-feed antimicrobials were used to raise 1 kg of live chicken and pig, respectively. Bacitracin (15.5% feeds), chlortetracycline (11.4%), and enramycin (10.8%) were the most common antimicrobials present in chicken feed formulations, whereas bacitracin (24.8%), chlortetracycline (23.9%), and florfenicol (17.4%) were the most common in pig feed formulations. Overall, 57% of the total quantitative usage consisted of antimicrobials regarded by WHO of importance for human medicine, including amoxicillin, colistin, tetracyclines, neomycin, lincomycin, and bacitracin. These figures confirm a very high magnitude of in-feed consumption of antimicrobials, especially in pig production. Results from this study should encourage further monitoring of antimicrobials used in animal production, and foster discussion about existing policies on inclusion of antimicrobials in animal feed rations.

Nhung NT, Thuy CT, Trung NV, Campbell J, Baker S, Thwaites G, Hoa NT, Carrique-Mas J. 2015. Induction of Antimicrobial Resistance in Escherichia coli and Non-Typhoidal Salmonella Strains after Adaptation to Disinfectant Commonly Used on Farms in Vietnam. Antibiotics (Basel), 4 (4), pp. 480-494. | Show Abstract | Read more

In Vietnam, commercial disinfectants containing quaternary ammonium compounds (QACs) are commonly used in pig and poultry farms to maintain hygiene during production. We hypothesized that sustained exposure to sub-bactericidal concentrations of QAC-based disinfectants may result in increased levels of antimicrobial resistance (AMR) among Enterobacteriacea due to the increase of efflux pump expression. To test this hypothesis we exposed six antimicrobial-susceptible Escherichia coli (E. coli) and six antimicrobial-susceptible non-typhoidal Salmonella (NTS) isolates to increasing concentrations of a commonly used commercial disinfectant containing a mix of benzalkonium chloride and glutaraldehyde. Over the 12-day experiment, strains exhibited a significant change in their minimum inhibitory concentration (MIC) of the disinfectant product (mean increase of 31% (SD ± 40)) (p = 0.02, paired Wilcoxon test). Increases in MIC for the disinfectant product were strongly correlated with increases in MIC (or decreases in inhibition zone) for all antimicrobials (Pearson's correlation coefficient 0.71-0.83, all p < 0.01). The greatest increases in MIC (or decreases in inhibition zone) were observed for ampicillin, tetracycline, ciprofloxacin, and chloramphenicol, and the smallest for gentamicin, trimethoprim/sulphamethoxazole. The treatment of 155 representative E. coli isolates from farmed and wild animals in the Mekong Delta (Vietnam) with phenyl-arginine beta-naphthylamide (PAβN), a generic efflux pump inhibitor, resulted in reductions in the prevalence of AMR ranging from 0.7% to 3.3% in these organisms, indicating a small contribution of efflux pumps on the observed prevalence of AMR on farms. These results suggest that the mass usage of commercial disinfectants, many of which contain QACs, is potentially a contributing factor on the generation and maintenance of AMR in animal production in Vietnam.

Loan HK, Cuong NV, Takhampunya R, Klangthong K, Osikowicz L, Kiet BT, Campbell J, Bryant J, Promstaporn S, Kosoy M et al. 2015. Bartonella species and trombiculid mites of rats from the Mekong Delta of Vietnam. Vector Borne Zoonotic Dis, 15 (1), pp. 40-47. | Show Abstract | Read more

A survey of Bartonella spp. from 275 rats purchased in food markets (n=150) and trapped in different ecosystems (rice field, forest, and animal farms) (n=125) was carried out during October, 2012-March, 2013, in the Mekong Delta of Vietnam. The overall Bartonella spp. prevalence detected by culture and PCR in blood was 14.9% (10.7-19.1%), the highest corresponding to Rattus tanezumi (49.2%), followed by Rattus norvegicus (20.7%). Trapped rats were also investigated for the presence and type of chiggers (larvae of trombiculid mites), and Bartonella spp. were investigated on chigger pools collected from each rat by RT-PCR. A total of five Bartonella spp. were identified in rats, three of which (B. elizabethae, B. rattimassiliensis, and B. tribocorum) are known zoonotic pathogens. Among trapped rats, factors independently associated with increased prevalence of Bartonella spp. included: (1) Rat species (R. tanezumi); (2) the number of Trombiculini-Blankaartia and Schoengastiini-Ascoschoengastia mites found on rats; and (3) the habitat of the rat (i.e., forest/fields vs. animal farms). The prevalence of Bartonella infection among chiggers from Bartonella spp.-positive R. tanezumi rats was 5/25 (25%), compared with 1/27 (3.7%) among Bartonella spp.-negative R. tanezumi rats (relative risk [RR]=5.4, 95% confidence interval [CI] 0.68-43.09). The finding of Bartonella spp.-positive chiggers on Bartonella spp.-negative rats is strongly suggestive of a transovarial transmission cycle. Rats are ubiquitous in areas of human activity and farms in the Mekong Delta; in addition, trapping and trading of rats for food is common. To correctly assess the human risks due to rat trapping, marketing, and carcass dressing, further studies are needed to establish the routes of transmission and cycle of infection. The widespread presence of these zoonotic pathogens in rats and the abundance of human-rat interactions suggest that surveillance efforts should be enhanced to detect any human cases of Bartonella infection that may arise.

Carrique-Mas JJ, Trung NV, Hoa NT, Mai HH, Thanh TH, Campbell JI, Wagenaar JA, Hardon A, Hieu TQ, Schultsz C. 2015. Antimicrobial usage in chicken production in the Mekong Delta of Vietnam. Zoonoses Public Health, 62 Suppl 1 (s1), pp. 70-78. | Show Abstract | Read more

Antimicrobials are used extensively in chicken production in Vietnam, but to date no quantitative data are available. A 2012-2013 survey of 208 chicken farms in Tien Giang province, stratified by size (10-200 chickens; >200-2000), was carried out to describe and quantify the use of antibacterial antimicrobials (usage per week per chicken and usage per 1000 chickens produced) in the Mekong Delta and to investigate factors associated with usage. Twenty-eight types of antimicrobial belonging to 10 classes were reported. Sixty-three per cent of all commercial formulations contained at least two antimicrobials. On 84% occasions, antimicrobials were administered with a prophylactic purpose. The overall adjusted quantities of antimicrobials used/week/chicken and per 1000 chickens produced (g) were 26.36 mg (SE ± 3.54) and 690.4 g (SE ± 203.6), respectively. Polypeptides, tetracyclines, penicillins and aminoglycosides were the antimicrobials used by most farms (18.6% farms, 17.5%, 11.3% and 10.1% farms, respectively), whereas penicillins, lincosamides, quinolones, and sulphonamides/trimethoprim were quantitatively the most used compounds (8.27, 5.2, 3.16 and 2.78 mg per week per chicken, respectively). Factors statistically associated with higher levels of usage (per week per chicken) were meat farms (OR = 1.40) and farms run by a male farmer (OR = 2.0). All-in-all-out farming systems (correlated with medium farms) were associated with reduced levels of antimicrobial usage (OR = 0.68). Usage levels to produced meat chickens were considerably higher than those reported in European countries. This should trigger the implementation of surveillance programmes to monitor sales of antimicrobials that should contribute to the rational administration of antimicrobials in order to preserve the efficacy of existing antimicrobials in Vietnam.

Carrique-Mas JJ, Bryant JE. 2013. A review of foodborne bacterial and parasitic zoonoses in Vietnam EcoHealth, 10 (4), pp. 465-489. | Show Abstract | Read more

Vietnam has experienced unprecedented economic and social development in recent years, and the livestock sector is undergoing significant transformations. Although food animal production is still dominated by small-scale 'backyard' enterprises with mixed crop-livestock or livestock-aquatic systems, there is a trend towards more intensive and vertically integrated operations. Changes in animal production, processing and distribution networks for meat and animal products, and the shift from wet markets to supermarkets will undoubtedly impact food safety risks in Vietnam in unforeseen and complex ways. Here, we review the available published literature on bacterial and parasitic foodborne zoonoses (FBZ) in Vietnam. We report on clinical disease burden and pathogen prevalence in animal reservoirs for a number of important FBZ, and outline opportunities for future research. © 2013 International Association for Ecology and Health.

Using mechanistic models to understand the impact of antimicrobial use in farming systems in Southeast Asia on antimicrobial resistance.

Antimicrobial resistance (AMR) is one of the most serious and rapidly growing public health threats in the world today and already accounts for hundreds of thousands of deaths each year. Widespread use of antibiotics in animal production systems is a significant concern for human health because of the associated risk of resistance and the fact that virtually all classes of antimicrobials important for human medicine are also used in animals. The use of antimicrobials and the generation of AMR ...

View project

2866