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Abstract Background In Plasmodium falciparum, resistance to chloroquine (CQ) is conferred by a K to T mutation at amino acid position 76 (K76T) in the P. falciparum CQ transporter (PfCRT). To date, at least 15 pfcrt genotypes, which are represented by combinations of five amino acids at positions 72-76, have been described in field isolates from various endemic regions. To identify novel mutant pfcrt genotypes and to reveal the genetic relatedness of pfcrt genotypes, a large-scale survey over a wide geographic area was performed. Methods Sequences for exon 2 in pfcrt, including known polymorphic sites at amino acid positions 72, 74, 75 and 76, were obtained from 256 P. falciparum isolates collected from eight endemic countries in Asia (Bangladesh, Cambodia, Lao P.D.R., the Philippines and Thailand), Melanesia (Papua New Guinea and Vanuatu) and Africa (Ghana). A haplotype network was constructed based on six microsatellite markers located -29 kb to 24 kb from pfcrt in order to examine the genetic relatedness among mutant pfcrt genotypes. Results In addition to wild type (CVMNK at positions 72-76), four mutant pfcrt were identified; CVIET, CVIDT, SVMNT and CVMNT (mutated amino acids underlined). Haplotype network revealed that there were only three mutant pfcrt lineages, originating in Indochina, Philippines and Melanesia. Importantly, the Indochina lineage contained two mutant pfcrt genotypes, CVIET (n = 95) and CVIDT (n = 14), indicating that CVIDT shares a common origin with CVIET. Similarly, one major haplotype in the Melanesian lineage contained two pfcrt genotypes; SVMNT (n = 71) and CVMNT (n = 3). In Africa, all mutant pfcrt genotypes were the CVIET of the Indochina lineage, probably resulting from the intercontinental migration of CQ resistance from Southeast Asia. Conclusions The number of CQ-mutant lineages observed in this study was identical to that found in previous studies. This supports the hypothesis that the emergence of novel CQ resistance is rare. However, in the mutant pfcrt genotypes, amino acid changes at positions 72, 74 and 75 appear to have recently been generated at least several times, producing distinct pfcrt mutant genotypes. The occurrence of new mutations flanking K76T may yield stronger resistance to CQ and/or a higher fitness than the original pfcrt mutant.

Several outbreaks of trichinellosis associated with the consumption of raw pork have occurred in Laos since 2004. This cross-sectional study was conducted in four provinces of northern Laos to investigate the seroepidemiology of trichinellosis in the human population and determine the prevalence and species of Trichinella infection in the domestic pig population. Serum samples and questionnaire data were obtained from 1419 individuals. Serum samples were tested for Trichinella antibodies by ELISA using larval excretory–secretory (ES) antigens and a subset of 68 positive samples were tested by western blot. The seroprevalence of Trichinella antibodies was 19.1% (95% confidence interval (CI) = 17.1–21.1%). The risk of having antibodies detected by ELISA using ES antigens increased with age, being of Lao-Tai ethnicity, living in Oudomxay province and being male. Tongue and diaphragm muscle samples were collected from 728 pigs and tested for Trichinella larvae by the artificial digestion method. Trichinella larvae were isolated from 15 pigs (2.1%) of which 13 were identified as T. spiralis by molecular typing; the species of the two remaining isolates could not be determined due to DNA degradation. Trichinella spp. are endemic in the domestic environment of northern Laos and targeted preventative health measures should be initiated to reduce the risk of further outbreaks occurring. [ABSTRACT FROM AUTHOR]

Objectives: To achieve malaria elimination by 2030, the Lao People's Democratic Republic (PDR) adopted a reactive surveillance and response (RASR) strategy of malaria case notification within 1 day, case investigation and classification within 3 days and foci investigation and response within 7 days. It is important to evaluate the performance and feasibility of RASR implementation in Lao PDR so that the strategy may be optimised and better contribute towards the goal of malaria elimination., Design: A mixed-methods study comprising of secondary data analysis of routinely collected malaria surveillance data, quantitative surveys and qualitative consultations was conducted in 2022., Setting: Primary data collections for quantitative surveys and qualitative consultations were conducted in Huaphan, Khammouane, Luangprabang and Savannakhet Provinces of Lao PDR., Participants: Quantitative surveys were conducted among malaria programme stakeholders and service providers. Qualitative interviews were conducted with malaria programme stakeholders, and focus group discussions with malaria programme stakeholders, service providers and mobile and migrant populations (MMPs)., Outcome Measures: Outcomes of interests were awareness and acceptability of current RASR activities by different group of participants, implementation, performance and feasibility of RASR activities including enablers and barriers., Results: In Lao PDR, malaria programme stakeholders and service providers were aware of RASR; however, these activities were not well known in MMPs. Respectively, the timeliness of case notification and case investigation was 0.0% and 15.6% in 2018 but increased to 98.0% and 98.6% in 2022. Implementation of RASR was acceptable to the malaria programme stakeholders and service providers, and continued implementation was perceived as feasible. Nevertheless, issues such as low level of community awareness, high level of migration and limitations in health system capacity were identified., Conclusion: Overall, the timeliness of case notification and investigation in Lao PDR was high, and malaria programme stakeholders and service providers had positive opinions on RASR. However, some operational and health system-related barriers were identified, which need to be addressed to improve the performance of RASR in Lao PDR., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

We assessed the insecticidal efficacy of Olyset nets after 5 years of use in rural villages of Lao PDR and evaluated the relationship between the physical condition of the nets and their insecticidal effect. Our results showed that most of the Olyset nets remained effective after 5 years of use; however, there was no significant relationship between the physical condition of the nets (ie, presence of holes, level of stains) and the insecticidal effect. The presence of large holes in polyester nets compared to the Olyset nets suggest the Olyset nets are stronger; however, nearly half of Olyset nets had small holes or had been previously repaired. Interestingly, the insecticide concentration and knockdown (KD) rate for 3 stored nets was low compared to the other nets routinely used in the house. To maintain the effectiveness of Olyset nets in rural villages of Lao PDR and other areas, residents should be advised to repair and store the nets appropriately and avoid exposure to high temperatures and direct sunlight for long periods.