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Fulltext Molecular characterization of Plasmodium falciparum antifolate resistance markers in Thailand between 2008 and 2016

Sugaram R, Suwannasin K, Kunasol C, Mathema VB, Day NPJ, Sudathip P, et al.

Malar J, 2020 Mar 04;19(1):107.
PMID: 32127009 DOI: 10.1186/s12936-020-03176-x

BACKGROUND: Resistance to anti-malarials is a major threat to the control and elimination of malaria. Sulfadoxine-pyrimethamine (SP) anti-malarial treatment was used as a national policy for treatment of uncomplicated falciparum malaria in Thailand from 1973 to 1990. In order to determine whether withdrawal of this antifolate drug has led to restoration of SP sensitivity, the prevalence of genetic markers of SP resistance was assessed in historical Thai samples.
METHODS: Plasmodium falciparum DNA was collected from the Thailand-Myanmar, Thailand-Malaysia and Thailand-Cambodia borders during 2008-2016 (N = 233). Semi-nested PCR and nucleotide sequencing were used to assess mutations in Plasmodium falciparum dihydrofolate reductase (pfdhfr), P. falciparum dihydropteroate synthase (pfdhps). Gene amplification of Plasmodium falcipaurm GTP cyclohydrolase-1 (pfgch1) was assessed by quantitative real-time PCR. The association between pfdhfr/pfdhps mutations and pfgch1 copy numbers were evaluated.
RESULTS: Mutations in pfdhfr/pfdhsp and pfgch1 copy number fluctuated overtime through the study period. Altogether, 14 unique pfdhfr-pdfhps haplotypes collectively containing quadruple to octuple mutations were identified. High variation in pfdhfr-pfdhps haplotypes and a high proportion of pfgch1 multiple copy number (51% (73/146)) were observed on the Thailand-Myanmar border compared to other parts of Thailand. Overall, the prevalence of septuple mutations was observed for pfdhfr-pfdhps haplotypes. In particular, the prevalence of pfdhfr-pfdhps, septuple mutation was observed in the Thailand-Myanmar (50%, 73/146) and Thailand-Cambodia (65%, 26/40) border. In Thailand-Malaysia border, majority of the pfdhfr-pfdhps haplotypes transaction from quadruple (90%, 9/10) to quintuple (65%, 24/37) during 2008-2016. Within the pfdhfr-pfdhps haplotypes, during 2008-2013 the pfdhps A/S436F mutation was observed only in Thailand-Myanmar border (9%, 10/107), while it was not identified later. In general, significant correlation was observed between the prevalence of pfdhfr I164L (ϕ = 0.213, p-value = 0.001) or pfdhps K540E/N (ϕ = 0.399, p-value ≤ 0.001) mutations and pfgch1 gene amplification.
CONCLUSIONS: Despite withdrawal of SP as anti-malarial treatment for 17 years, the border regions of Thailand continue to display high prevalence of antifolate and anti-sulfonamide resistance markers in falciparum malaria. Significant association between pfgch1 amplification and pfdhfr (I164L) or pfdhps (K540E) resistance markers were observed, suggesting a compensatory mutation.
Fulltext Zoonotic neglected tropical diseases at the animal-human interface in the Greater Mekong Subregion: Two decades of surveillance in Laos and Cambodia

Blacksell SD, Ward MP, Young JR, Bush RD, Windsor PA, Khounsy S, et al.

PLoS Negl Trop Dis, 2026 Jul;20(7):e0014584.
PMID: 42531271 DOI: 10.1371/journal.pntd.0014584

Zoonotic neglected tropical diseases (NTDs) remain a substantial but under-recognised source of human morbidity and economic concern in the Greater Mekong Subregion, particularly in settings characterised by close human-animal interaction. This article represents a narrative synthesis of zoonotic disease research conducted in Laos and Cambodia between 2000 and 2025. The review integrates published literature with findings from long-term surveillance programmes conducted by the authors and collaborating institutions in Laos, with comparative insights from Cambodia, to examine the presence, distribution, diversity, and drivers of zoonotic pathogens at the human-animal interface. Evidence demonstrates the endemic presence of a wide range of parasitic, bacterial, and viral zoonoses, including Taenia solium, Trichinella spp., Streptococcus suis, rickettsial infections, melioidosis, hepatitis E virus, and Japanese encephalitis virus. Some of these pathogens are sustained within smallholder livestock systems, informal slaughter, farming practises, food networks, and wet market environments, where limited diagnostic capacity and fragmented surveillance obscure true disease presence. Surveillance innovations, including abattoir-based sampling, cross-sectoral serological studies, environmental surveillance approaches, and molecular diagnostic tools, have improved pathogen detection but have also highlighted persistent structural and behavioural barriers to control. Socio-cultural practices, occupational exposure, wildlife trade, and economic dependencies reinforce transmission dynamics, indicating that biomedical interventions alone are insufficient. Instead, zoonotic disease persistence reflects the interaction of livestock production systems, environmental conditions, diagnostic limitations, and entrenched human behaviours. This review emphasises the need for integrated One Health approaches that combine strengthened surveillance, improved diagnostics, behavioural interventions, and regional collaboration. Addressing zoonotic NTDs in Laos and Cambodia requires coordinated strategies that account for both biological complexity and socio-economic context to achieve sustainable disease control and improved public health outcomes.

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