Stability of Genotube® Swabs for African Swine Fever Virus Detection Using Loop-Mediated Isothermal (LAMP) Laboratory Testing on Samples Stored without Refrigeration.

African swine fever (ASF) is a transboundary viral disease which causes high mortality in pigs. In many low- and middle-income countries and in remote areas where diagnostic surveillance for ASF virus (ASFV) is undertaken, access to trained animal health technicians, sample collection, cold chain storage and transport of samples to suitably equipped laboratories can be limiting when traditional sampling and laboratory tests are used. Previously published studies have demonstrated that alternative sampling matrices such as swabs and filter papers can be tested using PCR without refrigeration for up to a week. This study used Genotube® swabs stored in temperate and tropical climates without refrigeration for four weeks after collection to demonstrate there was no change in test performance and results using loop-mediated isothermal amplification (LAMP) ASFV detection on a series of pig serum samples including serum spiked with a synthetic ASFV positive control, naturally acquired ASFV positive serum from Timor-Leste and negative ASFV serum samples. The use of Genotube® swabs for ASFV detection for surveillance purposes, coupled with testing platforms such as LAMP, can provide an alternative to traditional testing methodology where resources are limited and time from collection to testing of samples is prolonged.

Prevalence of Antimicrobial Resistance in Escherichia coli and Salmonella Species Isolates from Chickens in Live Bird Markets and Boot Swabs from Layer Farms in Timor-Leste.

The rapid emergence of antimicrobial resistance is a global concern, and high levels of resistance have been detected in chicken populations worldwide. The purpose of this study was to determine the prevalence of antimicrobial resistance in Escherichia coli and Salmonella spp. isolated from healthy chickens in Timor-Leste. Through a cross-sectional study, cloacal swabs and boot swabs were collected from 25 live bird markets and two layer farms respectively. E. coli and Salmonella spp. from these samples were tested for susceptibility to six antimicrobials using a disk diffusion test, and a subset was tested for susceptibility to 27 antimicrobials using broth-based microdilution. E. coli and Salmonella spp. isolates showed the highest resistance towards either tetracycline or ampicillin on the disk diffusion test. E. coli from layer farms (odds ratio:5.2; 95%CI 2.0-13.1) and broilers (odds ratio:18.1; 95%CI 5.3-61.2) were more likely to be multi-drug resistant than those from local chickens. Based on the broth-based microdilution test, resistance to antimicrobials in the Timor-Leste Antimicrobial Guidelines for humans were low, except for resistance to ciprofloxacin in Salmonella spp. (47.1%). Colistin resistance in E. coli was 6.6%. Although this study shows that antimicrobial resistance in chickens was generally low in Timor-Leste, there should be ongoing monitoring in commercial chickens as industry growth might be accompanied with increased antimicrobial use.

Community engagement strengthens pig disease knowledge and passive surveillance in Timor-Leste.

Smallholder pig production in Timor-Leste is culturally and economically important for most households. However, regular and ongoing disease surveillance and pig husbandry training for farmers are limited. This article describes collaborative social and diagnostic research followed by a pilot community engagement program to improve farmer and technician knowledge, skills, and working relationships. There were three phases: (1) A qualitative study in 2020 to explore the experiences and knowledge of 133 pig farmers, 6 village leaders, and 16 district veterinary technicians on pig diseases and reporting, treatment methods, and access to information or assistance. (2) A pilot community engagement program in 3 villages in 2021 with the diagnostic investigation with samples analyzed from 27 dead pigs, and (3) Evaluation of community engagement and training outcomes. Results of the qualitative study revealed limited reporting of sick or dead pigs by farmers to veterinary technicians due to a lack of trust in the veterinary diagnostic system. Most technicians lacked experience with sampling or post-mortems so diagnostic training was undertaken for the pilot disease investigation. Evaluation results showed improved knowledge, motivation, and confidence of government staff and farmers. The credibility of veterinary technicians improved and gave them more confidence to work with communities. Farmers felt supported because all aspects of pig husbandry were addressed, and they were more willing to report dead or sick pigs. The project indicates that improved passive disease surveillance can be achieved by engaging communities in smallholder pig farming in Timor-Leste. Further research and testing of the approach in other districts and countries is recommended.

Antimicrobial Use in Animals in Timor-Leste Based on Veterinary Antimicrobial Imports between 2016 and 2019.

Monitoring veterinary antimicrobial use is part of the global strategy to tackle antimicrobial resistance. The purpose of this study was to quantify veterinary antimicrobials imported into Timor-Leste between 2016 and 2019 and describe the antimicrobial import profile of importers. Data were obtained from import applications received by the Ministry of Agriculture and Fisheries (MAF) of Timor-Leste. Import quantities were analysed by antimicrobial class, importance for human medicine, recommended route of administration and type of importer. An average of 57.4 kg (s.d. 31.0 kg) and 0.55 mg/kg (s.d. 0.27 mg/kg) animal biomass of antimicrobials was imported per year. Tetracyclines (35.5%), penicillins (23.7%), and macrolides (15.9%) were the commonly imported antimicrobial classes. Antimicrobials imported for parenteral administration were most common (60.1%). MAF was the largest importer (52.4%). Most of the critically important antimicrobials for human medicine were imported by poultry farms for oral administration and use for growth promotion could not be ruled out. In conclusion, the use of antimicrobials in animals in Timor-Leste is very low, in keeping with its predominantly subsistence agriculture system. Farmer education, development of treatment guidelines, and strengthening of the veterinary service is important for addressing the potential future misuse of antimicrobials especially in the commercial poultry industry.

Counting the cost: The potential impact of African Swine Fever on smallholders in Timor-Leste.

In September 2019, Timor-Leste, where pigs are kept by more than 70 percent of households, became the eleventh Asian country to report African Swine Fever (ASF). Drawing on our recent, as-yet unpublished research we show that while national pork consumption is low, pigs hold tremendous monetary value for smallholders within the economy of ceremonies. Given the sums of money paid for live pigs, the value of the national pig herd is around USD160 million - more than USD1000 per pig keeping household. Accordingly, pigs serve to buffer families against shocks and pressures, especially for health and education expenses. While not a zoonosis, the potential for ASF to lead to significant, negative impacts for smallholder farmers in Timor-Leste - some of the world's most vulnerable people - cannot be underestimated. We argue that Timor-Leste faces significant challenges in responding to ASF and there is a strong case for international support.