Understanding the veterinary antibiotics supply chain to address antimicrobial resistance in Lao PDR: Roles and interactions of involved stakeholders.

In response to the global call to mitigate risks associated with antimicrobial resistance (AMR), new regulations on the access and use of veterinary antibiotics are currently being developed by the Lao government. This study aims to explore how the implementation of these new regulations might effectively reduce and adapt the sale, distribution and use of veterinary antibiotics in Lao PDR. To this end, we used the theory of change, framing the AMR issue within the context of the stakeholders involved in the veterinary antibiotics supply chain. Qualitative and quantitative methods were used to collect data, based on questionnaires (n=36 antibiotic suppliers, n=96 chicken farmers, n=96 pig farmers), and participatory tools such as a workshop (n=10 participants), semi-structured interviews (n=20), and focus group discussions (n=7 participants). The stakeholders' understanding of the AMR issue and potential challenges related to the implementation of new regulations regarding access and use of antibiotics, were also investigated. We mapped the veterinary antibiotic supply chain in Lao PDR, and analysed the roles and interactions of its stakeholders. Twenty-three stakeholders representing the private and the public sectors were identified. Many informal and formal links connected these stakeholder within this supply chain. The lack of veterinarian-farmer interaction and the evolving nature of the veterinary antibiotics supply chain accentuated the challenges of achieving behaviour change through regulations. Most of the antibiotics found on farms were categorized by the World Health Organisation's as critically important antibiotics used in human medicine. We argue that AMR risk mitigation strategy requires dialogue and engagement between private and public sectors stakeholders, involved in the importation, distribution, sale and use of veterinary antibiotics. This study further highlighted that AMR is a complex adaptive challenge requiring multi-sectoral approach. We believed that a sustainable approach to reduce and adapt veterinary antibiotics use should be prepared in collaboration with stakeholders from private and public sectors identified in this study, in addition to the new regulations. This collaboration should start with the co-construction of a common understanding of AMR issue and of the objectives of new regulations.

Preliminary Study on Prevalence, Risk Factor and Genetic Homogeneity of Porcine Reproductive and Respiratory Syndrome Virus in Registered Pig Farms in Heilongjiang, China.

While porcine reproductive and respiratory syndrome (PRRS) causes great economic losses in southern and central China, systematic studies on the epidemiology of PRRS virus (PRRSV) in Heilongjiang Province had not been performed. Therefore, we conducted a preliminary study to estimate the prevalence and risk factors associated with PRRSV infection, as well as characterize the PRRSV in registered pig farms in Heilongjiang Province, China in 2011. A total of 1237 blood samples were collected from 72 farms and tested by reverse transcription polymerase chain reaction (RT-PCR) for PRRSV. Risk factors associated to PRRSV infection were analysed using logistic regression models. Genes of non-structural protein-2 (Nsp2) and glycoprotein 5 (GP5) from 22 isolates were sequenced for phylogenetic analysis. The results showed that the herd apparent prevalence was 9.7% (95% CI: 6.3, 13.1) in Heilongjiang Province. An increased risk of PRRSV infection on farms was associated with unrestricted movement of external people (OR = 14.1, 95% CI: 1.68, 119.07), close proximity (<1 km) to the nearest house, road or neighbouring farm (OR = 16.2, 95% CI: 1.52, 171.80), and selling farm products at both local and provincial markets (OR = 20.6, 95% CI: 2.02, 210.56). Phylogenetic analysis based on partial amino acid sequences of GP5 and Nsp2 showed that all the 22 PRRSV isolates in Heilongjiang are closely related to the highly pathogenic PRRSV strain JXA1 and belong to the Genotype 2 (American genotype). The prevalence, determination of risk factors and phylogenetic characterization will provide information for future epidemiological studies and a reference for developing surveillance and control strategies in this region.

Risk of Introduction in Northern Vietnam of HPAI Viruses from China: Description, Patterns and Drivers of Illegal Poultry Trade.

Poultry movement is known to contribute to the dissemination of highly pathogenic avian influenza (HPAI) viruses. In Northern Vietnam, the illegal trade of poultry from China is a source of concern and is considered as responsible for the regular introduction of new H5N1 viruses. The general objective of this study was to get a better understanding of this illegal trade (organization, volume, actors involved and drivers) to propose adequate preventive and control options. The information was also used to qualitatively evaluate the risk of exposure of susceptible poultry to HPAI H5N1 virus introduced from China by illegally traded poultry. We found that the main products imported from China are spent hens, day-old chicks (DOCs) and ducklings; spent hens being introduced in very large number. The drivers of this trade are multiple: economic (especially for spent hens) but also technical (demand for improved genetic potential for DOC and ducklings). Furthermore, these introductions also meet a high consumer demand at certain periods of the year. We also found that spatial dispersion of a batch of poultry illegally introduced from China is extensive and rapid, making any prediction of possible new outbreaks very hazardous. Finally, a risk mitigation plan should include measures to tackle the drivers of this trade or to legally organize it, to limit the threat to the local poultry sector. It is also essential for traders to be progressively better organized and biosecure and for hygienic practices to be enforced, as our study confirmed that at-risk behaviours are still very common among this profession.

How to reach the poor? Surveillance in low-income countries, lessons from experiences in Cambodia and Madagascar.

Surveillance of animal diseases in developing countries faces many constraints. Innovative tools and methods to enhance surveillance in remote and neglected areas should be defined, assessed and applied in close connection with local farmers, national stakeholders and international agencies. The authors performed a narrative synthesis of their own publications about surveillance in Madagascar and Cambodia. They analysed the data in light of their fieldwork experiences in the two countries' very challenging environments. The burden of animal and zoonotic diseases (e.g. avian influenza, African swine fever, Newcastle disease, Rift Valley fever) is huge in both countries which are among the poorest in the world. Being poor countries implies a lack of human and financial means to ensure effective surveillance of emerging and endemic diseases. Several recent projects have shown that new approaches can be proposed and tested in the field. Several advanced participatory approaches are promising and could be part of an innovative method for improving the dialogue among different actors in a surveillance system. Thus, participatory modelling, developed for natural resources management involving local stakeholders, could be applied to health management, including surveillance. Data transmission could benefit from the large mobile-phone coverage in these countries. Ecological studies and advances in the field of livestock surveillance should guide methods for enhancing wildlife monitoring and surveillance. Under the umbrella of the One Health paradigm, and in the framework of a risk-based surveillance concept, a combination of participatory methods and modern technologies could help to overcome the constraints present in low-income countries. These unconventional approaches should be merged in order to optimise surveillance of emerging and endemic diseases in challenging environments.

Evaluation of enzyme-linked immunosorbent assays and a haemagglutination inhibition tests for the detection of antibodies to Newcastle disease virus in village chickens using a Bayesian approach.

Newcastle disease (ND) is an endemic disease in village chickens in Ethiopia with substantial economic importance. The sensitivity (Se) and specificity (Sp) of a blocking enzyme-linked immunosorbent assay (bELISA, Svanova Biotech), indirect ELISA (iELISA, Laboratoire Service International) and haemagglutination inhibition (HI) test for ND virus (NDV) antibody detection were evaluated in a Bayesian framework in the absence of a gold standard test, on sera collected from unvaccinated chickens kept under the village production system in household flocks and at markets in two woredas (i.e. districts) of the Eastern Shewa zone, Ethiopia. The outcomes of the iELISA test differed dramatically from those of the two other tests with 92% of the samples testing positive as compared with less than 15% for bELISA and HI. iELISA results were also inconsistent with previous estimations of Newcastle serological prevalence. The information provided by the iELISA test was thus considered as highly unreliable, probably due to an extremely low specificity, and thus not considered in the Bayesian models aiming at estimating serological prevalence and test performance parameters. Bayesian modelling of HI and bELISA test results suggested that bELISA had both the highest Se (86.6%; 95% posterior credible interval (PCI): 61.8%; 98.5%), and the highest Sp (98.3%; 95% PCI: 97.2%; 99.5%), while HI had a Se of 80.2% (95% PCI: 59.1%; 94.3%), and a Sp of 96.1% (95% PCI: 95.1%; 97.4%). Model selection and the range of the posterior distribution of the correlation between bELISA and HI test outcomes for truly seropositive animals (median at 0.461; PCI: -0.055; 0.894) suggested a tendency for bELISA and HI to detect the same truly positive animals and to fail to detect the same truly positive animals. The use of bELISA in screening and surveillance for NDV antibodies is indicated given its high Se and Sp, in addition to its ease of automation to handle large numbers of samples compared to HI. The latter can be used as confirmatory test where an ELISA test with moderate or low specificity is used although the likely positive dependence with bELISA implies that HI and bELISA provide similar information on truly positive animals. Evaluation of commercial ELISAs is indicated before their wider use in village chicken populations to avoid erroneous inferences.

Epidemiological Analysis of Influenza A Infection in Cambodian Pigs and Recommendations for Surveillance Strategies.

This study analysed the available data of seroprevalence to human influenza viruses in pigs in Cambodia using generalized linear mixed models in order to improve understanding of factors underlying the spread of human influenza viruses in Cambodian pigs. The associations between seroprevalence against seasonal H1N1 influenza virus in pigs and the population density of humans and pigs were not significant. However, a positive association between anti-H3 antibodies in pigs and the human population density was identified. In contrast, there was a negative association between seroprevalence of H3N2 in pigs and the pig population density. Our study has highlighted the difficulty in identifying epidemiological risk factors when a limited data set is used for analyses. We therefore provide recommendations on data collection for future epidemiological analyses that could be improved by collecting metadata related to the animals sampled. In addition, serosurveillance for influenza A viruses in pigs in high-risk areas or at slaughterhouses is recommended in resource-limited countries.

Evaluating the efficiency of participatory epidemiology to estimate the incidence and impacts of foot-and-mouth disease among livestock owners in Cambodia.

The economic and social impacts of foot-and-mouth disease (FMD) for livestock owners of developed countries have been extensively documented over the past few years. In developing countries such as Cambodia, this evaluation is often lacking due to the scarcity of accurate data. In the present study, we used a range of participatory tools to infer farmers' knowledge and perception, and the relative incidence of FMD from January 2009 to June 2010 in fifty-one villages of Svay Rieng province, Cambodia. In addition, the detection of non-structural protein at village level was used to cross-validate the results from the participatory epidemiology (PE) study. A quantitative assessment using Bayesian modeling was carried out to assess the ability of PE to retrospectively determine the FMD-infected status of a village in Cambodia. Our study shows that even if FMD is ranked second in the list of priority diseases, livestock owners did not see any benefit in reporting it since the disease entailed low direct losses. The average clinical incidence rates at individual level for cattle-buffaloes and pigs in infected villages were assessed by proportional piling at 18% and 11%, respectively for the year 2009. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of PE study were estimated at 87%, 30%, 51% and 74%, respectively. This approach seems to largely overestimate the presence of the disease but proves useful in evaluating the impact of FMD at household level and in understanding the reasons for not reporting it. This information may be important in establishing well-adapted disease prevention and control strategies in Cambodia.

Seroprevalence of Newcastle disease and other infectious diseases in backyard chickens at markets in Eastern Shewa zone, Ethiopia.

This study was conducted to estimate the seroprevalence of Newcastle disease (ND), Pasteurella multocida (PM) infection, Mycoplasma gallisepticum (MG) infection, and infectious bursal disease (IBD) and to assess the level of concurrent seropositivity during the dry and wet seasons of the year 2010. In total, 234 and 216 sera were collected during the dry and wet seasons, respectively, from unvaccinated backyard chickens at 4 live poultry markets in 2 woredas (districts) of Eastern Shewa zone, Ethiopia, and were tested using commercial ELISA kits. The overall seroprevalence of ND, PM, MG, and IBD was 5.9, 66.2, 57.7, and 91.9%, respectively, during the dry season, and 6.0, 63.4, 78.7, and 96.3%, respectively, during the wet season. The seroprevalence of MG was higher (P < 0.001) during the wet season than during the dry season and higher (P = 0.002) in Adami-Tulu-Jido-Kombolcha woreda (74%) than in Ada'a woreda (60%). Area and season had no significant effect on the seroprevalence of ND, IBD, and PM, indicating the widespread presence of those pathogens throughout the year in the study area. Of all the chickens tested, 85.6% had antibodies concurrently to more than one of the pathogens investigated. Birds were concurrently seropositive to more diseases during the wet season (median = 3) than during the dry season (median = 2; P = 0.002). As serology is not able to distinguish between strains, further studies are warranted to better understand the circulating strains, their interactions, and their economic effect on backyard poultry production in Ethiopia.

Conceptual framework for avian influenza risk assessment in Africa: the case of Ethiopia.

The avian influenza (AI) epidemic is threatening Africa mainly because the flyways of migratory birds link the endemic and newly infected countries with disease-free areas in this continent and because of the risk of introduction through trade. Risk analysis provides a set of tools for supporting decision making by the veterinary services and other stakeholders, resulting in more effective surveillance and emergency preparedness. The risk assessment process could be split into three different steps: 1) risk release through the migratory birds and the official and unofficial poultry-product marketing chains; 2) risk exposure by means of studying interfaces among imported and exposed poultry and among wild and domestic birds; and 3) risk consequences for establishing the probability of AI spreading within the poultry population and the probability of it escaping detection. A conceptual framework is presented based on preliminary data and field missions carried out in Ethiopia. Field surveys and expert opinion will be necessary for the parameterization of the risk model. Spatial analysis will be used to identify high risk of exposure among wild and domestic birds. Risk communication and risk management will be based on the findings from the risk assessment model.

HPAI surveillance programme in Cambodia: results and perspectives.

Cambodia has faced 15 confirmed highly pathogenic avian influenza (H5N1) outbreaks in different sectors of the poultry industry since January 2004. The country has very limited human and financial resources and, when the outbreak first began, the veterinary services were not equipped with the basic tools to collect accurate epidemiological information or to fight the disease. Therefore, different agencies, under the umbrella of the Food and Agriculture Organisation, are providing support to the Government to strengthen its capacity to diagnose, survey and control the avian influenza (AI) virus. Different surveillance tools are being tested, such as market monitoring and a sentinel villages' network, to offset the weakness of the national passive surveillance network. Several constraints were identified during the implementation of this programme, such as a lack of motivation among provincial staff, the limited capacity of the central team to compile and analyse the data generated, the reluctance of farmers to have their animals sampled, and weak diagnostic capacities. The sustainability of such a surveillance system once international support ends remains to be seen. Participatory epidemiology (PE) may be an appropriate complementary tool to track diseases. PE works on the principle that livestock keepers often possess detailed knowledge of animal diseases and can provide valuable diagnostics that could help in identifying AI outbreaks, particularly in remote areas.