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Background: Pythiosis is a rare disease with high mortality, with over 94% of cases reported from Thailand and India. Prompt diagnosis and surgery improves patient outcomes. Therefore, continuing professional development (CPD) is essential for early recognition. However, a needs assessment related to a pythiosis CPD program has not been performed. Objectives: We conducted a needs assessment to develop a pythiosis CPD program. Patients/Methods: We conducted a survey study with 267 King Chulalongkorn Memorial Hospital residents (141 internal medicine (IM) residents and 126 surgery residents). A 30-item survey consisting of a knowledge assessment, demographic section, and an attitudes portion was distributed both electronically and via paper. The data was analyzed with descriptive and inferential statistics. Results: Sixty-seven percent completed the survey (110/141 IM residents, 70/126 surgery residents). The mean score [95% confidence interval] on the knowledge assessment was 41.67% [39.64%-43.69%] across all objectives. The three domains with the highest scores were pythiosis risk factors (67.22% correct), microbiologic characteristics (50.83%), and radiographic interpretation (50.56%). The three domains with the lowest scores were laboratory investigation (15.00%), epidemiology (29.17%), and symptomatology (30.83%). Most participants noted that the program should be online with both synchronous and asynchronous sessions, with a preferred length of 60–90 minutes per session. Conclusion: The pythiosis CPD program should emphasize education regarding symptomatology, laboratory investigation, and epidemiology, all of which are critical for the early detection of pythiosis to decrease mortality from this devastating disease. Most respondents felt this program was necessary and should be implemented in a virtual blended format. Author summary: Researchers conducted a needs assessment to develop a Continuing Professional Development (CPD) program on pythiosis, a rare and often fatal disease prevalent in Thailand and India. The study surveyed 267 residents at King Chulalongkorn Memorial Hospital, revealing a 67% completion rate. The residents exhibited a mean knowledge score of 41.67%, with the highest proficiency in pythiosis risk factors (67.22%), microbiologic characteristics (50.83%), and radiographic interpretation (50.56%). Conversely, lower scores were observed in laboratory investigation (15.00%), epidemiology (29.17%), and symptomatology (30.83%). Participants expressed a preference for an online CPD program with both synchronous and asynchronous sessions, each lasting 60–90 minutes. The study emphasizes the need for education on symptomatology, laboratory investigation, and epidemiology to enhance early pythiosis detection and reduce mortality. Respondents widely supported the implementation of the proposed virtual blended CPD program. [ABSTRACT FROM AUTHOR]

An estimated 105 million dengue infections occur per year across 120 countries, where traditional vector control is the primary control strategy to reduce contact between mosquito vectors and people. The ongoing sars-cov-2 pandemic has resulted in dramatic reductions in human mobility due to social distancing measures; the effects on vector-borne illnesses are not known. Here we examine the pre and post differences of dengue case counts in Malaysia, Singapore and Thailand, and estimate the effects of social distancing as a treatment effect whilst adjusting for temporal confounders. We found that social distancing is expected to lead to 4.32 additional cases per 100,000 individuals in Thailand per month, which equates to 170 more cases per month in the Bangkok province (95% CI: 100–242) and 2008 cases in the country as a whole (95% CI: 1170–2846). Social distancing policy estimates for Thailand were also found to be robust to model misspecification, and variable addition and omission. Conversely, no significant impact on dengue transmission was found in Singapore or Malaysia. Across country disparities in social distancing policy effects on reported dengue cases are reasoned to be driven by differences in workplace-residence structure, with an increase in transmission risk of arboviruses from social distancing primarily through heightened exposure to vectors in elevated time spent at residences, demonstrating the need to understand the effects of location on dengue transmission risk under novel population mixing conditions such as those under social distancing policies. Author summary: Significant healthcare burdens arise from dengue and traditional vector control is the primary control strategy to reduce dengue transmission potential. The ongoing sars-cov-2 pandemic has resulted in dramatic reductions in human mobility due to social distancing measures; the effects on vector-borne illnesses are not known. This paper examines the pre and post differences of dengue case counts in Malaysia, Singapore and Thailand, and estimate the effects of social distancing as a treatment effect whilst accounting for temporal confounders. We found that social distancing is expected to lead to an increase in the number of cases in Thailand, with the largest increase arising from Bangkok province. Social distancing policy estimates for Thailand were also found to be robust in sensitivity analysis. Conversely, no significant impact on dengue transmission was found in Singapore or Malaysia. Across country disparities in social distancing policy effects on reported dengue cases are reasoned to be driven by differences in workplace-residence structure, with an increase in transmission risk of arboviruses from social distancing primarily through heightened exposure to vectors in elevated time spent at residences, demonstrating the need to understand the effects of location on dengue transmission risk under novel population mixing conditions such as those under social distancing policies. [ABSTRACT FROM AUTHOR]

In 2018–2019, Thailand experienced a nationwide spread of chikungunya virus (CHIKV), with approximately 15,000 confirmed cases of disease reported. Here, we investigated the evolutionary and molecular history of the East/Central/South African (ECSA) genotype to determine the origins of the 2018–2019 CHIKV outbreak in Thailand. This was done using newly sequenced clinical samples from travellers returning to Sweden from Thailand in late 2018 and early 2019 and previously published genome sequences. Our phylogeographic analysis showed that before the outbreak in Thailand, the Indian Ocean lineage (IOL) found within the ESCA, had evolved and circulated in East Africa, South Asia, and Southeast Asia for about 15 years. In the first half of 2017, an introduction occurred into Thailand from another South Asian country, most likely Bangladesh, which subsequently developed into a large outbreak in Thailand with export to neighbouring countries. Based on comparative phylogenetic analyses of the complete CHIKV genome and protein modelling, we identified several mutations in the E1/E2 spike complex, such as E1 K211E and E2 V264A, which are highly relevant as they may lead to changes in vector competence, transmission efficiency and pathogenicity of the virus. A number of mutations (E2 G205S, Nsp3 D372E, Nsp2 V793A), that emerged shortly before the outbreak of the virus in Thailand in 2018 may have altered antibody binding and recognition due to their position. This study not only improves our understanding of the factors contributing to the epidemic in Southeast Asia, but also has implications for the development of effective response strategies and the potential development of new vaccines. Author summary: We investigated the evolutionary and molecular history of the East/Central/South African (ECSA) genotype to determine the origins of the 2018–2019 chikungunya virus (CHIKV) outbreak in Thailand. We used newly sequenced clinical samples from travellers returning to Sweden from Thailand in late 2018 and early 2019 together with previously published genome sequences. Our phylogeographic analysis shows that the Indian Ocean lineage (IOL), found within ECSA, evolved in Eastern Africa, Southern Asia, and Southeast Asia for about 15 years before the outbreak in Thailand in 2018. We have also identified amino acid substitutions that may be associated with immune evasion, increased spread, and higher virulence that occurred prior to the outbreak and may have played a critical role in the rapid spread of the virus. Our study concludes that monitoring and understanding CHIKV dynamics remains critical for an effective response to the previously unpredictable outbreaks of the virus. [ABSTRACT FROM AUTHOR]

Background: Soil-transmitted helminthiases (STHs) are common in tropical and subtropical regions. Southern Thailand experiences an extended rainy season, leading to persistently moist soil. This condition supports the life cycle of STHs, hindering effective control due to reinfection and low drug efficacy. We implemented a novel STH control strategy during the dry season aimed at decreasing reinfection rates without enhancing sanitation or hygiene practices. However, there were unexpected, prolonged droughts linked to El Niño events from 2014 to 2016. Additionally, we assessed the effects of these drought conditions on further control measures without the use of anthelmintics. Methodology/Principal findings: A longitudinal study was conducted from 2012 to 2016. Stool samples collected from 299 participants were analyzed using the Kato-Katz and agar plate culture methods. Participants who tested positive for STHs received a single 400 mg dose of albendazole. The efficacy of the treatment was evaluated three weeks later. To confirm the control measures were implemented during the dry season, we monitored the number of rainy days following albendazole treatment for 52 days, of which 38 were without rain. Follow-up stool examinations were carried out in 2013 and 2016, with no additional doses of albendazole administered. Rainfall and rainy day data, which served as indicators of unexpected droughts due to El Niño, were collected from the nearest local meteorological stations. Before the drought, there was a decrease in STH prevalence in 2013—except for trichuriasis—attributable to the dry season control efforts. Despite these efforts, STH prevalence remained high. Remarkably, in 2016, following the drought period, the prevalence of trichuriasis, which had not changed previously, spontaneously declined without further albendazole treatment compared to 2013. Furthermore, the prevalence of strongyloidiasis remained unchanged likely due to its low susceptibility to drought conditions, as it can reproduce within hosts. Conversely, the prevalence of other STHs consistently declined. The drought and possible improvements in sanitation and hygiene practices contributed to this decrease by reducing rates of reinfection and new infection and by increasing the natural cure rate. Additionally, some participants infected with hookworms or Trichuris who were not cured by albendazole experienced natural remission. Conclusions/Significance: Control measures implemented during the dry season, combined with a 14-month-long drought induced by the El Niño event of 2014–2016, and some improvements in sanitation and hygiene practices, contributed to a decrease in both the prevalence and intensity of STHs, except for S. stercoralis. Over time, S. stercoralis is likely to become the predominant species among the STHs. Author summary: Soil-transmitted helminths (STHs) are prevalent in tropical rainforest regions of southern Thailand, where frequent rainfall can enhance the environmental life cycle of STHs, leading to reinfection. To mitigate this, we implemented control measures during the dry season from 2012 to 2016, aiming to reduce reinfection rates. For STH detection, we used agar plate culture and the Kato-Katz methods to analyze stool samples collected from 299 participants. Only those who tested positive for STHs received a single dose of albendazole, without accompanying health education, sanitation, or hygiene interventions. The effectiveness of the treatment was evaluated three weeks later. Follow-up assessments were conducted in 2013 and 2016 without further administration of albendazole. In 2013, a year after the albendazole treatment, there was a decrease in the prevalence of all STHs, except for trichuriasis. Unexpectedly, the El Niño event from 2014–2016 triggered prolonged droughts from 2014 to 2015, which likely led to a reduction in the prevalence and intensity of all STHs, excluding S. stercoralis. This exception was due to its low natural cure rate (NCR), as it can reproduce within the host. Additionally, the drought likely led to a reduction in the reinfection rate and an increase in the NCR. Notably, the NCR of trichuriasis induced by drought was more significant than that achieved through albendazole treatment. Furthermore, participants infected with hookworm or T. trichiura who were not cured by albendazole experienced natural remission. Improvements in sanitation and hygiene also likely contributed to the decreased prevalence of STHs. Based on these findings, we recommend that the control of STHs, especially in tropical rainforests characterized by long rainy seasons and persistently wet soil, should be strategically timed during the dry season to optimize effectiveness. [ABSTRACT FROM AUTHOR]

Background: Malaria transmission-blocking vaccines (TBVs) aim to inhibit malaria parasite development in mosquitoes and prevent further transmission to the human host. The putative-secreted ookinete protein 25 (PSOP25), highly conserved in Plasmodium spp., is a promising TBV target. Here, we investigated PvPSOP25 from P. vivax as a TBV candidate using transgenic murine parasite P. berghei and clinical P. vivax isolates. Methods and findings: A transgenic P. berghei line expressing PvPSOP25 (TrPvPSOP25Pb) was generated. Full-length PvPSOP25 was expressed in the yeast Pichia pastoris and used to immunize mice to obtain anti-rPvPSOP25 sera. The transmission-blocking activity of the anti-rPvPSOP25 sera was evaluated through in vitro assays and mosquito-feeding experiments. The antisera generated by immunization with rPvPSOP25 specifically recognized the native PvPSOP25 antigen expressed in TrPvPSOP25Pb ookinetes. In vitro assays showed that the immune sera significantly inhibited exflagellation and ookinete formation of the TrPvPSOP25Pb parasite. Mosquitoes feeding on mice infected with the transgenic parasite and passively transferred with the anti-rPvPSOP25 sera showed a 70.7% reduction in oocyst density compared to the control group. In a direct membrane feeding assay conducted with five clinical P. vivax isolates, the mouse anti-rPvPSOP25 antibodies significantly reduced the oocyst density while showing a negligible influence on mosquito infection prevalence. Conclusions: This study supported the feasibility of transgenic murine malaria parasites expressing P. vivax antigens as a useful tool for evaluating P. vivax TBV candidates. Meanwhile, the moderate transmission-reducing activity of the generated anti-rPvPSOP25 sera necessitates further research to optimize its efficacy. Author summary: The persistence of Plasmodium vivax poses a significant public health concern in certain regions, particularly Southeast Asia. The distinctive biology of Plasmodium vivax presents challenges for its control and eradication efforts. The development of a transmission-blocking vaccine is considered an essential strategy for malaria elimination, while the identification of antigen candidates plays a critical role in vaccine development. In this study, we aimed to evaluate the potential of PvPSOP25 as a transmission-blocking vaccine candidate using the transgenic murine parasite P. berghei. The transmission-reducing activity of anti-rPvPSOP25 sera was evaluated by both in vitro and mosquito-feeding experiments. Additionally, a direct membrane feeding assay using clinical P. vivax isolates from Thailand further validated the moderate transmission-reducing activity exhibited by the anti-rPvPSOP25 antisera. [ABSTRACT FROM AUTHOR]

Background: Dengue is spreading in (sub)tropical areas, and half of the global population is at risk. The macroeconomic impact of dengue extends beyond healthcare costs. This study evaluated the impact of dengue on gross domestic product (GDP) based on approaches tailored to two dengue-endemic countries, Thailand and Brazil, from the tourism and workforce perspectives, respectively. Findings: Because the tourism industry is a critical economic sector for Thailand, lost tourism revenues were estimated to analyze the impact of a dengue outbreak. An input-output model estimated that the direct effects (on international tourism) and indirect effects (on suppliers) of dengue on tourism reduced overall GDP by 1.43 billion US dollars (USD) (0.26%) in the outbreak year 2019. The induced effect (reduced employee income/spending) reduced Thailand's GDP by 375 million USD (0.07%). Overall, lost tourism revenues reduced Thailand's GDP by an estimated 1.81 billion USD (0.33%) in 2019 (3% of annual tourism revenue). An inoperability input-output model was used to analyze the effect of workforce absenteeism on GDP due to a dengue outbreak in Brazil. This model calculates the number of lost workdays associated with ambulatory and hospitalized dengue. Input was collected from state-level epidemiological and economic data for 2019. An estimated 22.4 million workdays were lost in the employed population; 39% associated with the informal sector. Lost workdays due to dengue reduced Brazil's GDP by 876 million USD (0.05%). Conclusions: The economic costs of dengue outbreaks far surpass the direct medical costs. Dengue reduces overall GDP and inflicts national economic losses. With a high proportion of the population lacking formal employment in both countries and low income being a barrier to seeking care, dengue also poses an equity challenge. A combination of public health measures, like vector control and vaccination, against dengue is recommended to mitigate the broader economic impact of dengue. Author summary: Dengue is a viral illness spread by mosquitoes in Southeast Asia and South America, and about half of the world's population is at risk. Burden-of-illness studies on the economic impact of dengue typically focus on medical costs. However, the impact of this infectious disease extends far beyond healthcare costs. This study evaluated the impact of dengue on gross domestic product (GDP) based on approaches tailored to two dengue-endemic countries, Thailand and Brazil. The analyses estimated that the dengue outbreak in 2019 had a profound impact on Thailand's tourism industry, reducing GDP by an estimated US$1.81 billion, equivalent to 3% of annual tourism revenues or 0.33% of total GDP. Dengue also impacted Brazil's workforce in 2019, leading to an estimated 22.4 million lost workdays, of which 39% were associated with the informal sector, and reducing GDP overall by US$876 million (0.05% of total GDP). Informal workers are vulnerable to dengue-related financial burdens, and a low income can be a barrier to seeking medical care for dengue. This study highlights the broader economic impact of dengue beyond healthcare costs and underscores the importance of a combination of public health measures, such as vector control and vaccination. [ABSTRACT FROM AUTHOR]

Targeted environmental and ecosystem management remain crucial in control of dengue. However, providing detailed environmental information on a large scale to effectively target dengue control efforts remains a challenge. An important piece of such information is the extent of the presence of potential dengue vector breeding sites, which consist primarily of open containers such as ceramic jars, buckets, old tires, and flowerpots. In this paper we present the design and implementation of a pipeline to detect outdoor open containers which constitute potential dengue vector breeding sites from geotagged images and to create highly detailed container density maps at unprecedented scale. We implement the approach using Google Street View images which have the advantage of broad coverage and of often being two to three years old which allows correlation analyses of container counts against historical data from manual surveys. Containers comprising eight of the most common breeding sites are detected in the images using convolutional neural network transfer learning. Over a test set of images the object recognition algorithm has an accuracy of 0.91 in terms of F-score. Container density counts are generated and displayed on a decision support dashboard. Analyses of the approach are carried out over three provinces in Thailand. The container counts obtained agree well with container counts from available manual surveys. Multi-variate linear regression relating densities of the eight container types to larval survey data shows good prediction of larval index values with an R-squared of 0.674. To delineate conditions under which the container density counts are indicative of larval counts, a number of factors affecting correlation with larval survey data are analyzed. We conclude that creation of container density maps from geotagged images is a promising approach to providing detailed risk maps at large scale. [ABSTRACT FROM AUTHOR]

Background: Montelukast has shown potential as a candidate treatment for dengue. This study aimed to evaluate the efficacy and safety of montelukast in preventing dengue with warning signs. Methods: This multicenter, randomized, double-blind, placebo-controlled trial enrolled adult participants with NS1 antigenemia in Thailand. The participants were randomly assigned to receive either oral montelukast (10 mg) or a placebo for 10 days or until all symptoms resolved. Results: Between January 2021 and June 2023, 358 participants were enrolled and randomly assigned (1:1) to receive either montelukast or placebo. The incidence rate of warning signs in the montelukast group and the placebo group were 9.5% and 7.8% per day, respectively. There was no difference between the two groups (HR 1.36; 95%CI 0.94–1.96, P = 0.105). No statistically significant differences were observed in the incidence rate of severe dengue, hemoconcentration, thrombocytopenia, admission, or recovery from dengue. Neither dengue shock, nor mortality occurred. The montelukast group exhibited a decreased incidence rate of transaminase elevations (0.7% vs 1.4% per day, HR: 0.48, 95%CI 0.25–0.90, P = 0.023). Conclusion: Oral montelukast does not reduce the incidence of warning signs among patients with dengue. Nevertheless, the observed decrease in transaminase elevations warrants further investigation to evaluate the potential effect of montelukast. Clinical Trials Registration: Clinicaltrials.gov, NCT04673422, registered on 9 December 2020 Author summary: Although there is no specific treatment for dengue to date, montelukast shows promise as a potential therapeutic agent. An open-label study previously showed a 71% reduction in dengue shock among patients with dengue who were administered montelukast. However, concerns were raised regarding the clinical study's lack of a power analysis or predefined statistical difference. Because of its affordability, relatively safe profile, and widespread availability, exploring its role in preventing dengue complications holds great significance. This study is a double-blind clinical trial with pragmatic design assessing the efficacy of montelukast in dengue. In contrast to the previous open-label study, montelukast does not reduce the incidence of warning signs among patients with dengue. Nevertheless, a reduction in the incidence of transaminase elevations was observed. In conclusion, montelukast should not be used for the prevention of warning signs in patients with dengue. However, the observed decrease in transaminase elevations warrants further investigation to evaluate the potential effect of montelukast. [ABSTRACT FROM AUTHOR]

Dengue is a mosquito-borne disease that has spread to over 100 countries. Its symptoms vary from the relatively mild acute febrile illness called dengue fever to the much more severe dengue shock syndrome. Dengue is caused by dengue virus (DENV), which belongs to the Flavivirus genus of the family Flaviviridae. There are four serotypes of DENV, i.e., DENV1 to DENV4, and each serotype is divided into distinct genotypes. Thailand is an endemic area where all four serotypes of DENV co-circulate. Genome sequencing of the DENV2 that was isolated in Thailand in 2016 and 2017 revealed the emergence of the Cosmopolitan genotype and its co-circulation with the Asian-I genotype. However, it was unclear whether different genotypes have different levels of viral replication and pathogenicity. Focus-forming assay (FFA) results showed that clinical isolates of these genotypes differed in focus size and proliferative capacity. Using circular polymerase extension reaction, we generated parental and chimeric viruses with swapped genes between these two DENV2 genotypes, and compared their focus sizes and infectivity titers using FFA. The results showed that the focus size was larger when the structural proteins and/or non-structural NS1-NS2B proteins were derived from the Cosmopolitan virus. The infectious titers were consistent with the focus sizes. Single-round infectious particle assay results confirmed that chimeric viruses with Cosmopolitan type structural proteins, particularly prM/E, had significantly increased luciferase activity. Replicon assay results showed that Cosmopolitan NS1-NS2B proteins had increased reporter gene expression levels. Furthermore, in interferon-receptor knock-out mice, viruses with Cosmopolitan structural and NS1-NS2B proteins had higher titers in the blood, and caused critical disease courses. These results suggested that differences in the sequences within the structural and NS1-NS2B proteins may be responsible for the differences in replication, pathogenicity, and infectivity between the Asian-I and Cosmopolitan viruses. Author summary: Since the discovery of dengue virus (DENV) in 1907, many people in Latin America, and South and Southeast Asia have been at risk of infection. Analysis of the sequences of viruses isolated in Thailand in 2016 and 2017 revealed the emergence of the Cosmopolitan genotype of DENV type 2 (DENV2), and its co-circulation with the pre-existing Asian-I genotype of DENV2. This co-circulation still exists in Thailand. We noted that there were large differences in the infectious titers between these genotypes, and the focus sizes also differed. In the present study, we analyzed both the in vitro and in vivo characteristics of these viruses using multiple recombinant viruses to determine which viral genetic region is responsible for these differences. We identified a genetic region encoding structural and NS1-NS2B proteins that affects the infectivity, replication, and pathogenicity of the viruses. Our study provides new insights into the transmission of new DENVs, and the interactions between DENV proteins. [ABSTRACT FROM AUTHOR]

In 1971, scientists from Mahidol University in Thailand and the Smithsonian Institution in the USA formed a research team to study a new species of Schistosoma in the Mekong River in Thailand and Laos. The studies, completed during 1971–1973, prior to the construction of any dams or restrictions to the natural flow regime of the Mekong River, provide a unique description of the natural ecological state of the river that can serve as a baseline for current research. The natural transmission of Schistosoma japonicum, Mekong Strain, was first reported on Khong Island, Laos in 1973 using sentinel mice. The first detailed description of the habitat ecology of the snail vector Neotricula aperta was done on-site in 1971 simultaneously with that research and is unique in providing the only description of the river shoreline habitat before any dams were built and any alteration of the natural flow regime was in place. Aggregating current information in a Place-Based Conceptual Model (PBCM) as an organizing template, along with current habitat models that combine ecological data with e-flows, can be developed and used as a tool to predict suitable habitats for snails. The natural flow regime of the Mekong River prior to any impoundments is described with current updates on the potential impacts of climate change and dams with flow-related snail habitat characteristics, including sediment drift and water quality. The application of the PBCM to describe and compare descriptive information on current and potential future N. aperta/S. mekongi habitat is discussed. Author summary: We provide a unique profile of Neotricula aperta habitat in the Mekong River in Laos and Thailand during the transmission of Schistosoma mekongi prior to the construction of any dams on the river. A Place-Based Conceptual Model (PBCM) is described to help identify potential new areas, with suitable habitats for snails and disease transmission. The natural flow regime of the Mekong River prior to any impoundments is described with current updates on the potential impacts of climate change and flow-related snail habitat characteristics, including sediment drift and water quality. Environmental management strategies are outlined, emphasizing environmental flow (e-flow) and habitat models, with the potential to mitigate N. aperta distribution and S. mekongi disease transmission. The PBCM can serve as a template to aggregate and compare historical baseline information (Before dams) with current data (After dams) to create predictive models describing suitable habitats for N. aperta and other snail intermediate hosts and vectors along shoreline areas of the Mekong River continuum. The PBCM model could be a powerful management tool when combined with habitat suitability and e-flow models. Adding schistosome biomarkers currently under development in a One Health context to select possible areas of a river likely to support disease transmission could extend the value of the model. [ABSTRACT FROM AUTHOR]

Background: In South-East Asia, Thailand is the country with the highest number of human autochthonous cases of leishmaniases mostly due to Leishmania martiniquensis. Their transmission remains unresolved to date even though sand flies are known vectors of leishmaniases. As such, we focused a study on the sand fly fauna of a cave in Thailand to explore the biodiversity of potential Leishmania vectors. Main results: We carried out an inventory in Pha Tong cave. We caught and identified 570 Phlebotomine sand flies (452 females and 118 males) and identified 14 species belonging to the genera Phlebotomus, Idiophlebotomus, Chinius, Sergentomyia and Grassomyia. Among these 14 species, two could not be related to known sand fly species. Herein, we propose the description of two new sand fly species, previously unknown to science. The first new species, Phlebotomus shadenae n. sp. is a sand fly of the subgenus Anaphlebotomus. It is morphologically close to Ph. stantoni, a species widely distributed throughout Southeast Asia. However, it differs by the length of the genital filaments in males or by the length of the ducts of the spermathecae in females as well as the high divergence of cytochrome b sequences. Additionally, we revised the systematics of the subgenus Anaphlebotomus and reinstated, by examination of its holotype, the validity of Ph. maynei, an Indian wrongly considered as a synonym of Ph. stantoni in the past. The second new species, Sergentomyia maiae n. sp., differs from a species in the same group, Se. barraudi, by an original cibarial double row of vertical teeth as well as by molecular data. Conclusions: We propose the description of two new sand fly species for Science with morphological and molecular evidence. Ph. shadenae n. sp. was also found to be distributed in the south of Thailand and in Laos. Future studies need to determine whether these two species can play a role as vectors of Leishmania parasites, Trypanosomatids or Phlebovirus. Most of the species caught in the present study are strictly cavernicolous except Grassomyia sp. and a few Sergentomyia. Author summary: Phlebotomine sand flies are tiny blood sucking insects. These are vectors of Leishmania, flagellate unicellular parasites, mostly found in Mediterranean and tropical areas. In Thailand, Leishmania martiniquensis and L. orientalis are responsible for local diseases but the parasite transmission cycle remains unknown. For a long time, South-Eastern Asia was considered to be free of leishmaniasis and as such, Phlebotomine sand flies have been understudied in this area. We carried out an inventory in a cave located in the Chiang Rai district, northern Thailand. We caught two new species we consider to be new for Science based on morphological (head and genital apparatus) and molecular characters (cytochrome b mitochondrial DNA sequences). The new species were named Phlebotomus shadenae n. sp. and Sergentomyia maiae n. sp. We also recorded the first species in Laos whereas the second has just been recorded in the studied cave. [ABSTRACT FROM AUTHOR]

: A literature survey and analysis was conducted to describe the epidemiology of dengue disease in Thailand reported between 2000 and 2011. The literature search identified 610 relevant sources, 40 of which fulfilled the inclusion criteria defined in the review protocol. Peaks in the number of cases occurred during the review period in 2001, 2002, 2008 and 2010. A shift in age group predominance towards older ages continued through the review period. Disease incidence and deaths remained highest in children aged ≤15 years and case fatality rates were highest in young children. Heterogeneous geographical patterns were observed with higher incidence rates reported in the Southern region and serotype distribution varied in time and place. Gaps identified in epidemiological knowledge regarding dengue disease in Thailand provide several avenues for future research, in particular studies of seroprevalence. Protocol registration: PROSPERO [ABSTRACT FROM AUTHOR]

Background: A global stockpile of oral cholera vaccine (OCV) was established in 2013 for use in outbreak response and are licensed as two-dose regimens. Vaccine availability, however, remains limited. Previous studies have found that a single dose of OCV may provide substantial protection against cholera. Methods: Using a mathematical model with two age groups paired with optimization algorithms, we determine the optimal vaccination strategy with one and two doses of vaccine to minimize cumulative overall infections, symptomatic infections, and deaths. We explore counterfactual vaccination scenarios in three distinct settings: Maela, the largest refugee camp in Thailand, with high in- and out-migration; N'Djamena, Chad, a densely populated region; and Haiti, where departments are connected by rivers and roads. Results: Over the short term under limited vaccine supply, the optimal strategies for all objectives prioritize one dose to the older age group (over five years old), irrespective of setting and level of vaccination coverage. As more vaccine becomes available, it is optimal to administer a second dose for long-term protection. With enough vaccine to cover the whole population with one dose, the optimal strategies can avert up to 30% to 90% of deaths and 36% to 92% of symptomatic infections across the three settings over one year. The one-dose optimal strategies can avert 1.2 to 1.8 times as many cases and deaths compared to the standard two-dose strategy. Conclusions: In an outbreak setting, speedy vaccination campaigns with a single dose of OCV is likely to avert more cases and deaths than a two-dose pro-rata campaign under a limited vaccine supply. Author summary: Cholera is a major global burden, with 21,000–143,000 deaths annually. The oral cholera vaccine (OCV) is given as two doses and is less effective in children under five years old than in those aged five years and older. However, supply is limited. In this study, we used mathematical models paired with optimization algorithms to find the optimal strategies to allocate vaccines in order to minimize the number of cases and deaths for three distinct settings in Chad, Thailand, and Haiti. We found that in the short term (1 year) when there is limited vaccine supply, it is optimal to vaccinate individuals over five years old with one dose and young children under five years old with two doses. Across the three settings, these optimal strategies prevent the most cases, save the most lives, and avert 1.2 to 1.8 times as many cases and deaths as the standard two-dose untargeted strategy. Our results support that if vaccine supply is limited, under an outbreak setting, mass vaccination campaigns with a single dose of OCV may prevent more cases and save more lives than would a standard two-dose untargeted campaign. [ABSTRACT FROM AUTHOR]

Burkholderia pseudomallei is a soil-dwelling bacterium endemic to Southeast Asia and northern Australia that causes the disease, melioidosis. Although the global genomic diversity of clinical B. pseudomallei isolates has been investigated, there is limited understanding of its genomic diversity across small geographic scales, especially in soil. In this study, we obtained 288 B. pseudomallei isolates from a single soil sample (~100g; intensive site 2, INT2) collected at a depth of 30cm from a site in Ubon Ratchathani Province, Thailand. We sequenced the genomes of 169 of these isolates that represent 7 distinct sequence types (STs), including a new ST (ST1820), based on multi-locus sequence typing (MLST) analysis. A core genome SNP phylogeny demonstrated that all identified STs share a recent common ancestor that diverged an estimated 796–1260 years ago. A pan-genomics analysis demonstrated recombination between clades and intra-MLST phylogenetic and gene differences. To identify potential differential virulence between STs, groups of BALB/c mice (5 mice/isolate) were challenged via subcutaneous injection (500 CFUs) with 30 INT2 isolates representing 5 different STs; over the 21-day experiment, eight isolates killed all mice, 2 isolates killed an intermediate number of mice (1–2), and 20 isolates killed no mice. Although the virulence results were largely stratified by ST, one virulent isolate and six attenuated isolates were from the same ST (ST1005), suggesting that variably conserved genomic regions may contribute to virulence. Genomes from the animal-challenged isolates were subjected to a bacterial genome-wide association study to identify genomic regions associated with differential virulence. One associated region is a unique variant of Hcp1, a component of the type VI secretion system, which may result in attenuation. The results of this study have implications for comprehensive sampling strategies, environmental exposure risk assessment, and understanding recombination and differential virulence in B. pseudomallei. Author summary: Burkholderia pseudomallei is the causative agent of melioidosis, a disease endemic to Southeast Asia and northern Australia. The evaluation of diversity within B. pseudomallei has largely been conducted utilizing clinical isolates despite almost all infections emerging from environmental exposure. In this study, we surveyed the genomic diversity of 169 isolates collected from a single soil sample in Ubon Ratchathani Province, Thailand. Seven different sequence types were identified, and substantial within-sequence-type gene diversity was observed. To test for differential virulence, 30 isolates were challenged in a mouse melioidosis model. A small number of isolates killed all mice, but most killed none, demonstrating the variable virulence potential of different B. pseudomallei isolates present in the single sample. A comparative genomics analysis identified multiple genes associated with virulence, including Hcp1, a component of the type VI secretion system and a known virulence factor. The results of this study have implications for the comprehensive environmental surveillance, environmental exposure, and differential virulence of B. pseudomallei. [ABSTRACT FROM AUTHOR]

Background: Dengue is a major public health problem in Thailand, but data are often focused on certain dengue-endemic areas. Methods: To better understand dengue epidemiology and clinical characteristics in Thailand, a fever surveillance study was conducted among patients aged 1–55 years, who presented with non-localized febrile illness at Bang Phae Community Hospital in Ratchaburi province, Thailand from October 2011 to September 2016. Results: Among 951 febrile episodes, 130 were dengue-confirmed. Individuals aged 10–14 years were mostly affected, followed by those 15–19 years-of-age, with about 15% of dengue-confirmed cases from adults 25 years and older. There were annual peaks of dengue occurrence between June-November. Most prevalent serotype in circulation was DENV-2 in 2012, DENV-3 in 2014, and DENV-4 & -3 in 2015. Among dengue cases, 65% were accurately detected using the dengue NS1 RDT. Detection rate was similar between secondary and primary dengue cases where 66% of secondary vs. 60% of primary dengue cases had positive results on the NS1 RDT. Among dengue cases, 66% were clinically diagnosed with suspected dengue or DHF, prior to lab confirmation. Dengue was positively associated with rash, headache, hematemesis and alterations to consciousness, when compared to non-dengue. Dengue patients were 10.6 times more likely to be hospitalized, compared to non-dengue cases. Among dengue cases, 95 were secondary and 35 were primary infections. There were 8 suspected DHF cases and all were identified to be secondary dengue. Secondary dengue cases were 3.5 times more likely to be hospitalized compared to primary dengue cases. Although the majority of our dengue-positive patients were secondary dengue cases, with few patients showing manifestations of DHF, our dengue cases were mostly mild disease. Even among children < 10 years-of-age, 61% had secondary infection and the rate of secondary infection increased with age. Conclusion: While the majority of dengue-confirmed cases were children, almost three-quarters of dengue-confirmed cases in this study were secondary dengue. Our study results consistent with previous data from the country confirm the hyperendemic transmission of DENV in Thailand, even in the non-epidemic years. With various interventions becoming available for dengue prevention and control, including dengue vaccines, decision-making on future implementation strategies should be based on such burden of disease data. Author summary: Dengue is a well-known public health problem in Thailand. To identify epidemiologic and clinical patterns of dengue in Thailand, we studied 951 febrile patients with fever, aged 1–55 years, who attended Bang Phae district hospital in Ratchaburi province, Thailand. Patients were tested with a rapid test for dengue, and further tests were carried out on paired blood samples taken 10–21 days apart. Overall, 14% of the febrile episodes identified between October 2011 and September 2016 were dengue-confirmed. Teenagers were mostly affected with only few adults. Dengue peaks occurred between June-November, but there was no large epidemic during the study period. DENV type 2 was the main serotype in circulation in 2012, DENV type 3 in 2014, and DENV types 4 and 3 in 2015. Dengue cases were 11 times more likely to be hospitalized than non-dengue cases. Dengue patients were more likely to present with rash, headache, hematemesis and alterations to consciousness, compared to non-dengue. Two thirds of the dengue cases were clinically diagnosed with suspected dengue or DHF, even without lab confirmation. Secondary dengue cases were about three times more common than primary cases. Even among children < 10 years-of-age, 61% of the patients were of secondary infection. Among dengue cases, 65% were accurately detected using the dengue RDT. This was not different between secondary and primary dengue cases. All 8 patients clinically diagnosed with DHF were secondary dengue cases. Although most of our dengue-positive patients were secondary dengue cases, we only observed few patients showing DHF. Thus, we concluded that our dengue cases were mostly of mild dengue illness. Our study results support the previous data confirming the high burden of dengue in Thailand and such information would help to make decisions on future implementation of various tools for dengue prevention and control. [ABSTRACT FROM AUTHOR]

Background: Aedes mosquitoes are vectors for several major arboviruses of public health concern including dengue viruses. The relationships between Aedes infestation and disease transmission are complex wherein the epidemiological dynamics can be difficult to discern because of a lack of robust and sensitive indicators for predicting transmission risk. This study investigates the use of anti-Aedes saliva antibodies as a serological biomarker for Aedes mosquito bites to assess small scale variations in adult Aedes density and dengue virus (DENV) transmission risk in northeastern Thailand. Individual characteristics, behaviors/occupation and socio-demographics, climatic and epidemiological risk factors associated with human-mosquito exposure are also addressed. Methods: The study was conducted within a randomized clustered control trial in Roi Et and Khon Kaen provinces over a consecutive 19 months period. Thirty-six (36) clusters were selected, each of ten houses. Serological and entomological surveys were conducted in all houses every four months and monthly in three sentinel households per cluster between September 2017 and April 2019 for blood spot collections and recording concurrent immature and adult Aedes indices. Additionally, the human exposure to Aedes mosquito bites (i.e., Mosquito Exposure Index or MEI) was estimated by ELISA measuring levels of human antibody response to the specific Nterm-34 kDa salivary antigen. The relationships between the MEI, vector infestation indices (adult and immature stages) and vector DENV infection were evaluated using a two-level (house and individual levels) mixed model analysis with one-month lag autoregressive correlation. Results: There was a strong positive relationship between the MEI and adult Aedes (indoor and outdoor) density. Individuals from households with a medium mosquito density (mean difference: 0.091, p<0.001) and households with a high mosquito density (mean difference: 0.131, p<0.001) had higher MEI's compared to individuals from households without Aedes. On a similar trend, individuals from households with a low, medium or high indoor Aedes densities (mean difference: 0.021, p<0.007, 0.053, p<0.0001 and 0.037, p<0.0001 for low, medium and high levels of infestation, respectively) had higher MEI than individuals from houses without indoor Aedes., The MEI was driven by individual characteristics, such as gender, age and occupation/behaviors, and varied according to climatic, seasonal factors and vector control intervention (p<0.05). Nevertheless, the study did not demonstrate a clear correlation between MEI and the presence of DENV-infected Aedes. Conclusion: This study represents an important step toward the validation of the specific IgG response to the Aedes salivary peptide Nterm-34kDa as a proxy measure for Aedes infestation levels and human-mosquito exposure risk in a dengue endemic setting. The use of the IgG response to the Nterm-34 kDa peptide as a viable diagnostic tool for estimating dengue transmission requires further investigations and validation in other geographical and transmission settings. Author summary: Aedes mosquitoes and the viruses they transmit are major public health concerns for over half of the global human population. However, the quantitative relationships between virus transmission and vector mosquito infestation remain unclear despite numerous indicators used to estimate transmission risk and predict dengue outbreaks. The aim of this study is to investigate the use of a salivary biomarker to assess the small-scale variation in human exposure to Aedes bites and the risk of dengue infection in the context of a vector control intervention in northeastern Thailand. A cohort of 539 persons visited every four months, including 161 individuals visited monthly, were recruited for routine serological and concurrent household entomological surveys during 19 consecutive months follow-up. Antibody response to Aedes bites was measured by enzyme-linked immunosorbent assays to assess the mosquito exposure index (MEI) and association with the Aedes adult and immature abundance as well as the presence of dengue virus (DENV) in adult mosquitoes (transmission risk). Additionally, the individual (cohort), climatic, and vector control intervention risk factors associated with MEI are explored. This study demonstrates that the MEI was strongly related to household adult Aedes density, particularly indoors resting mosquitoes. Additionally, the MEI was influenced by individual characteristics (i.e., person age, gender, staying indoors), and varied according to seasons and intervention. Nonetheless, no clear relationship between MEI and dengue transmission risk (i.e., vector infection) was detected. This study demonstrated the potential usefulness of the MEI to assess heterogeneity in adult Aedes infestation indices that could assist public health authorities to rapidly identify mosquito "hot spots" and the timeliness of effective vector control interventions. [ABSTRACT FROM AUTHOR]

Dengue is an emerging vector-borne viral disease across the world. The primary dengue mosquito vectors breed in containers with sufficient water and nutrition. Outdoor containers can be detected from geotagged images using state-of-the-art deep learning methods. In this study, we utilize such container information from street view images in developing a risk mapping model and determine the added value of including container information in predicting dengue risk. We developed seasonal-spatial models in which the target variable dengue incidence was explained using weather and container variable predictors. Linear mixed models with fixed and random effects are employed in our models to account for different characteristics of containers and weather variables. Using data from three provinces of Thailand between 2015 and 2018, the models are developed at the sub-district level resolution to facilitate the development of effective targeted intervention strategies. The performance of the models is evaluated with two baseline models: a classic linear model and a linear mixed model without container information. The performance evaluated with the correlation coefficients, R-squared, and AIC shows the proposed model with the container information outperforms both baseline models in all three provinces. Through sensitivity analysis, we investigate the containers that have a high impact on dengue risk. Our findings indicate that outdoor containers identified from street view images can be a useful data source in building effective dengue risk models and that the resulting models have potential in helping to target container elimination interventions. Author summary: The primary dengue mosquito vectors breed in containers with sufficient water and nutrition. Outdoor containers can be detected from geotagged images using state-of-the-art deep learning methods. Eight breeding site container types in Google street view images are detected using convolutional neural networks. We investigate the added value of including container information from geotagged images in predicting dengue risk. To explain the target variable dengue incidence, weather variables are added to complement the container variable predictors. Linear mixed-effects models are built to account for the effects of spatial and seasonal variation in weather and container variables on the dengue incidence. Evaluation is carried out over three provinces in Thailand: Bangkok, Nakhon Si Thammarat, and Krabi in comparison with classic linear models as well as the mixed effect models without container information. The proposed model with the container information outperforms both baseline models in all three provinces. We further perform sensitivity analysis to investigate the sensitivity of dengue incidence to the changes in the number of containers as well as the improvement in the model performance. This is the first work on dengue risk prediction models using container density information from geotagged images analysis. [ABSTRACT FROM AUTHOR]

Background: Antibody responses as serological markers of Plasmodium vivax infection have been shown to correlate with exposure, but little is known about the other factors that affect antibody responses in naturally infected people from endemic settings. To address this question, we studied IgG responses to novel serological exposure markers (SEMs) of P. vivax in three settings with different transmission intensity. Methodology: We validated a panel of 34 SEMs in a Peruvian cohort with up to three years' longitudinal follow-up using a multiplex platform and compared results to data from cohorts in Thailand and Brazil. Linear regression models were used to characterize the association between antibody responses and age, the number of detected blood-stage infections during follow-up, and time since previous infection. Receiver Operating Characteristic (ROC) analysis was used to test the performance of SEMs to identify P. vivax infections in the previous 9 months. Principal findings: Antibody titers were associated with age, the number of blood-stage infections, and time since previous P. vivax infection in all three study sites. The association between antibody titers and time since previous P. vivax infection was stronger in the low transmission settings of Thailand and Brazil compared to the higher transmission setting in Peru. Of the SEMs tested, antibody responses to RBP2b had the highest performance for classifying recent exposure in all sites, with area under the ROC curve (AUC) = 0.83 in Thailand, AUC = 0.79 in Brazil, and AUC = 0.68 in Peru. Conclusions: In low transmission settings, P. vivax SEMs can accurately identify individuals with recent blood-stage infections. In higher transmission settings, the accuracy of this approach diminishes substantially. We recommend using P. vivax SEMs in low transmission settings pursuing malaria elimination, but they are likely to be less effective in high transmission settings focused on malaria control. Author summary: Plasmodium vivax still poses a threat in many countries due to its ability to cause recurrent infections. Key to achieving the goal of malaria elimination is the ability to quickly detect and treat carriers of relapsing parasites. Failing to identify this transmission reservoir will hinder progress towards malaria elimination. Recently, novel serological markers of recent exposure to P. vivax (SEM) have been developed and validated in low transmission settings. It is still poorly understood what factors affect the antibody response to these markers when evaluated in contrasting endemic contexts. To determine the factors that influence the antibody response to SEM, we compared the antibody levels in three sites with different transmission intensity: Thailand (low), Brazil (moderate) and Peru (high). In this study, we found that transmission intensity plays a key role in the acquisition of the antibody repertoire to P. vivax. In highly endemic sites, it is likely that immunological memory resulting from a constant and sustained exposure will impact the performance of SEMs to detect individuals with recent exposure to P. vivax. In summary, SEMs that perform well in low transmission sites do not perform as well in high transmission regions. [ABSTRACT FROM AUTHOR]

Human head lice are blood-sucking insects causing an infestation in humans called pediculosis capitis. The infestation is more prevalent in the school-aged population. Scalp itching, a common presenting symptom, results in scratching and sleep disturbance. The condition can lead to social stigmatization which can lead to loss of self-esteem. Currently, the mainstay of treatment for pediculosis is chemical insecticides such as permethrin. The extended use of permethrin worldwide leads to growing pediculicide resistance. The aim of this study is to demonstrate the presence of the knockdown resistance (kdr) mutation in head lice populations from six different localities of Thailand. A total of 260 head lice samples in this study were collected from 15 provinces in the 6 regions of Thailand. Polymerase chain reaction (PCR) was used to amplify the α subunit of voltage-sensitive sodium channel (VSSC) gene, kdr mutation (C→T substitution). Restriction fragment length polymorphism (RFLP) patterns and sequencing were used to identify the kdr T917I mutation and demonstrated three genotypic forms including homozygous susceptible (SS), heterozygous genotype (RS), and homozygous resistant (RR). Of 260 samples from this study, 156 (60.00%) were SS, 58 (22.31%) were RS, and 46 (17.69%) were RR. The overall frequency of the kdr T917I mutation was 0.31. Genotypes frequencies determination using the exact test of Hardy-Weinberg equilibrium found that northern, central, northeastern, southern, and western region of Thailand differed from expectation. The five aforementioned localities had positive inbreeding coefficient value (Fis > 0) which indicated an excess of homozygotes. The nucleotide and amino acid sequences of RS and RR showed T917I and L920F point mutations. In conclusion, this is the first study detecting permethrin resistance among human head lice from Thailand. PCR-RFLP is an easy technique to demonstrate the kdr mutation in head louse. The data obtained from this study would increase awareness of increasing of the kdr mutation in head louse in Thailand. Author summary: Head louse infestation is still a common health problem worldwide. Permethrin is the first line treatment of choice for pediculosis. However, the increase of recurrent cases and treatment failure of pediculosis raises public health concerns about the growing resistance of permethrin among head lice populations. Early determination of the permethrin resistance status is crucial in order to choose the pediculicide of choice appropriately. Our study demonstrated that the PCR-RFLP is a beneficial technique for knockdown resistance detection of T917I point mutation in head lice because it was inexpensive, accurate, and reproducible. Ultimately, our study emphasized that prompt investigation for permethrin resistance should be taken into account in the context of recurrent or treatment failure cases of pediculosis and the alternative chemical pediculicides based on different mechanism of action should be considered under those particular circumstances. [ABSTRACT FROM AUTHOR]

Background and aims: Dengue fever is a major public health problem in tropical/subtropical regions. Prior economic analyses have predominantly evaluated either vaccination or vector-control programmes in isolation and do not really consider the incremental benefits and cost-effectiveness of mixed strategies and combination control. We estimated the cost-effectiveness of single and combined approaches in Thailand. Methods: The impacts of different control interventions were analysed using a previously published mathematical model of dengue epidemiology and control incorporating seasonality, age structure, consecutive infection, cross protection, immune enhancement and combined vector-host transmission. An economic model was applied to simulation results to estimate the cost-effectiveness of 4 interventions and their various combinations (6 strategies): i) routine vaccination of 1-year olds; ii) chemical vector control strategies targeting adult and larval stages separately; iii) environmental management/ public health education and awareness [EM/ PHEA]). Payer and societal perspectives were considered. The health burden of dengue fever was assessed using disability-adjusted life-years (DALYs) lost. Costs and effects were assessed for 10 years. Costs were discounted at 3% annually and updated to 2013 United States Dollars. Incremental cost-effectiveness analysis was carried out after strategies were rank-ordered by cost, with results presented in a table of incremental analysis. Sensitivity and scenario analyses were undertaken; and the impact and cost-effectiveness of Wolbachia was evaluated in exploratory scenario analyses. Results: From the payer and societal perspectives, 2 combination strategies were considered optimal, as all other control strategies were dominated. Vaccination plus adulticide plus EM/ PHEA was deemed cost-effective according to multiple cost-effectiveness criteria. From the societal perspective, incremental differences vs. adulticide and EM/ PHEA resulted in costs of $157.6 million and DALYs lost of 12,599, giving an expected ICER of $12,508 per DALY averted. Exploratory scenario analyses showed Wolbachia to be highly cost-effective ($343 per DALY averted) vs. other single control measures. Conclusions: Our model shows that individual interventions can be cost-effective, but that important epidemiological reductions and economic impacts are demonstrated when interventions are combined as part of an integrated approach to combating dengue fever. Exploratory scenario analyses demonstrated the potential epidemiological and cost-effective impact of Wolbachia when deployed at scale on a nationwide basis. Our findings were robust in the face of sensitivity analyses. Author summary: Dengue fever has become a major public health problem. It is considered one of the most important mosquito-borne viral diseases and occurs in >100 countries in tropical and subtropical regions of Asia-Pacific, the Americas, the Middle East, and Africa with >3 billion people at risk. Despite current control interventions against dengue fever in endemic countries, the disease is associated with considerable healthcare utilisation, personal costs to patients and caregivers, productivity loss and human suffering. Whilst the illness is well understood, there is also recognition that current control efforts focussing predominantly on Aedes aegypti control and elimination are less than optimal although may still have an important role to play in the short to medium term. In this study, we consider the cost-effectiveness of individual as well as mixed dengue control strategies in Thailand, embracing chemical interventions, public health education/ environmental control and paediatric vaccination using a dengue vaccine profile broadly consistent with (dengue) vaccines in late stage development. To anticipate the transition to possible new vector control technologies, we also carry out exploratory scenario analyses of the impact and cost-effectiveness of the release of Wolbachia-infected mosquitoes (which are less capable of spreading viruses). Our findings indicate that single dengue control interventions can be cost-effective weapons in reducing dengue infections, although their effectiveness may be additionally enhanced when combined. [ABSTRACT FROM AUTHOR]

Background/Objectives: Dengue fever is an important public health concern in most tropical and subtropical countries, and its prevention and control rest on vector surveillance and control. However, many aspects of dengue epidemiology remain unclear; in particular, the relationship between Aedes vector abundance and dengue transmission risk. This study aims to identify entomological and immunological indices capable of discriminating between dengue case and control (non-case) houses, based on the assessment of candidate indices, as well as individual and household characteristics, as potential risk factors for acquiring dengue infection. Methods: This prospective, hospital-based, case-control study was conducted in northeastern Thailand between June 2016 and August 2019. Immature and adult stage Aedes were collected at the houses of case and control patients, recruited from district hospitals, and at patients' neighboring houses. Blood samples were tested by RDT and PCR to detect dengue cases, and were processed with the Nterm-34 kDa salivary peptide to measure the human immune response to Aedes bites. Socioeconomic status, and other individual and household characteristics were analyzed as potential risk factors for dengue. Results: Study findings showed complex relationships between entomological indices and dengue risk. The presence of DENV-infected Aedes at the patient house was associated with 4.2-fold higher odds of dengue. On the other hand, Aedes presence (irrespective of infectious status) in the patient's house was negatively associated with dengue. In addition, the human immune response to Aedes bites, was higher in control than in case patients and Aedes adult abundance and immature indices were higher in control than in case houses at the household and the neighboring level. Multivariable analysis showed that children aged 10–14 years old and those aged 15–25 years old had respectively 4.5-fold and 2.9-fold higher odds of dengue infection than those older than 25 years. Conclusion: DENV infection in female Aedes at the house level was positively associated with dengue infection, while adult Aedes presence in the household was negatively associated. This study highlights the potential benefit of monitoring dengue viruses in Aedes vectors. Our findings suggest that monitoring the presence of DENV-infected Aedes mosquitoes could be a better indicator of dengue risk than the traditional immature entomological indices. Author summary: Dengue fever is a globally expanding arboviral disease, consisting of four distinct serotypes, transmitted primarily by synanthropic/peridomestic mosquitoes, Aedes aegypti and Aedes albopictus. Given the absence of specific treatment, and the incomplete protection provided by the currently available vaccine, vector surveillance and control remain the principal tool to prevent and control dengue transmission. However, vector surveillance through the monitoring of larval mosquito indices lacks consistency in addressing dengue risk. Surveillance based on pupal and adult stages is considered as more accurate to estimate dengue transmission risk, although monitoring is difficult to implement in routine. An alternative strategy is the use of the specific human antibody response to Aedes saliva to identify human exposure risk to Aedes bites. We conducted a hospital-based, case-control study in northeastern Thailand in order to identify risk factors for dengue infection using entomological and immunological indices, together with select individual and household characteristics. We found that people aged 10–25 years had significant higher odds of dengue than older adults (>25 years old). The presence of DENV-infected Aedes in the house was associated with 4.2-fold higher odds of dengue infection. Interestingly, Aedes adult abundance in the household was negatively associated with dengue revealing the complex role of Aedes density to dengue risk. This study highlights the potential benefit of monitoring dengue viruses in Aedes vectors to identify areas ("hot spots") and people ("hot pops") at higher risk of transmission. [ABSTRACT FROM AUTHOR]

Important arboviral diseases, such as dengue, chikungunya, and Zika virus infections, are transmitted mainly by the Aedes aegypti vector. So far, controlling this vector species with current tools and strategies has not demonstrated sustainable and significant impacts. Our main objective was to evaluate whether open field release of sterile males, produced from combining the sterile insect technique using radiation with the insect incompatible technique through Wolbachia-induced incompatibility (SIT/IIT), could suppress natural populations of Ae. aegypti in semi-rural village settings in Thailand. Irradiated Wolbachia-infected Aedes aegypti males produced by the SIT/IIT approach were completely sterile and were able to compete with the wild fertile ones. Open field release of these sterile males was conducted in an ecologically isolated village in Chachoengsao Province, eastern Thailand. House-to-house visit and media reports resulted in community acceptance and public awareness of the technology. During intervention, approximately 100–200 sterile males were released weekly in each household. After 6 months of sterile male release, a significant reduction (p<0.05) of the mean egg hatch rate (84%) and the mean number of females per household (97.30%) was achieved in the treatment areas when compared to the control ones. Our study represents the first open field release of sterile Ae. aegypti males developed from a combined SIT/IIT approach. Entomological assessment using ovitraps, adult sticky traps, and portable vacuum aspirators confirmed the success in reducing natural populations of Ae. aegypti females in treated areas. Public awareness through media resulted in positive support for practical use of this strategy in wider areas. Further study using a systematic randomized trial is needed to determine whether this approach could have a significant impact on the diseases transmitted by Ae. aegypti vector. Dengue, chikungunya, and Zika are important diseases that pass to humans only by the bites of Aedes aegypti mosquitoes infected with these viruses. Due to the lack of an effective vaccine and drug to cure these diseases, mosquito control is the only method available to reduce disease risk. Most of the available preventive and control measures involve an application of chemical-based products. The inappropriate use of chemicals causes insecticide resistance and environmental contamination. In this study, we report on the pilot trial of an alternative, safe, and environmental friendly approach to suppress Ae. aegypti mosquito populations. This alternative method involves the release of sterile Ae. aegypti males that were infected with, naturally-occurring, insect bacteria Wolbachia, and were irradiated at low doses to obtain full sterilization. The pilot trial involved the release of 100–250 sterile males per household in a treated area over a 6-month period. High efficacy of this approach was evidenced from a significant reduction in the numbers of wild Ae. aegypti females and an increase in sterility by reduction in the egg hatch rate of wild Ae. aegypti females in the treated area. Public awareness through media resulted in public support to conduct trials in wider areas. [ABSTRACT FROM AUTHOR]

Introduction: The prevalence of bacteremia caused by Gram negative non-fermentative (GNNF) bacteria has been increasing globally over the past decade. Many studies have investigated their epidemiology but focus on the common GNNF including Pseudomonas aeruginosa and Acinetobacter baumannii. Knowledge of the uncommon GNNF bacteremias is very limited. This study explores invasive bloodstream infection GNNF isolates that were initially unidentified after testing with standard microbiological techniques. All isolations were made during laboratory-based surveillance activities in two rural provinces of Thailand between 2006 and 2014. Methods: A subset of GNNF clinical isolates (204/947), not identified by standard manual biochemical methodologies were run on the BD Phoenix automated identification and susceptibility testing system. If an organism was not identified (12/204) DNA was extracted for whole genome sequencing (WGS) on a MiSeq platform and data analysis performed using 3 web-based platforms: Taxonomer, CGE KmerFinder and One Codex. Results: The BD Phoenix automated identification system recognized 92% (187/204) of the GNNF isolates, and because of their taxonomic complexity and high phenotypic similarity 37% (69/187) were only identified to the genus level. Five isolates grew too slowly for identification. Antimicrobial sensitivity (AST) data was not obtained for 93/187 (50%) identified isolates either because of their slow growth or their taxa were not in the AST database associated with the instrument. WGS identified the 12 remaining unknowns, four to genus level only. Conclusion: The GNNF bacteria are of increasing concern in the clinical setting, and our inability to identify these organisms and determine their AST profiles will impede treatment. Databases for automated identification systems and sequencing annotation need to be improved so that opportunistic organisms are better covered. [ABSTRACT FROM AUTHOR]

Background: In 2015, Singapore had the first and only reported foodborne outbreak of invasive disease caused by the group B Streptococcus (GBS; Streptococcus agalactiae). Disease, predominantly septic arthritis and meningitis, was associated with sequence type (ST)283, acquired from eating raw farmed freshwater fish. Although GBS sepsis is well-described in neonates and older adults with co-morbidities, this outbreak affected non-pregnant and younger adults with fewer co-morbidities, suggesting greater virulence. Before 2015 ST283 had only been reported from twenty humans in Hong Kong and two in France, and from one fish in Thailand. We hypothesised that ST283 was causing region-wide infection in Southeast Asia. Methodology/Principal findings: We performed a literature review, whole genome sequencing on 145 GBS isolates collected from six Southeast Asian countries, and phylogenetic analysis on 7,468 GBS sequences including 227 variants of ST283 from humans and animals. Although almost absent outside Asia, ST283 was found in all invasive Asian collections analysed, from 1995 to 2017. It accounted for 29/38 (76%) human isolates in Lao PDR, 102/139 (73%) in Thailand, 4/13 (31%) in Vietnam, and 167/739 (23%) in Singapore. ST283 and its variants were found in 62/62 (100%) tilapia from 14 outbreak sites in Malaysia and Vietnam, in seven fish species in Singapore markets, and a diseased frog in China. Conclusions: GBS ST283 is widespread in Southeast Asia, where it accounts for a large proportion of bacteraemic GBS, and causes disease and economic loss in aquaculture. If human ST283 is fishborne, as in the Singapore outbreak, then GBS sepsis in Thailand and Lao PDR is predominantly a foodborne disease. However, whether transmission is from aquaculture to humans, or vice versa, or involves an unidentified reservoir remains unknown. Creation of cross-border collaborations in human and animal health are needed to complete the epidemiological picture. [ABSTRACT FROM AUTHOR]

Background: Burkholderia pseudomallei is the causative agent of melioidosis, a severe infectious disease in tropical regions. It is necessary to understand the risk of acquiring this infection from the environment. Methodology /Principal Findings: The prevalence, concentration and genetic diversity of B. pseudomallei isolates collected from two sites in Buriram, Northeast Thailand were investigated. Forty-four environmental samples (18 from soil, 14 from rice rhizosphere, and 12 from water) were collected; of those 44 samples, 19 were collected from near a patient’s residence and 25 from suspected exposure sites and compared with 10 clinical isolates of the patient. Quantitative culture was performed, and B. pseudomallei was identified using the latex agglutination test and matrix-laser absorption ionisation mass spectrometry. Genotyping was performed in 162 colonies from clinical (N = 10) and environmental samples (N = 152) using pulse-field gel electrophoresis (PFGE) followed by multi-locus sequence typing (MLST) of the clinical strain. B. pseudomallei was detected in 11 of the 44 environmental samples (1 from soil, 4 from rice rhizosphere, and 6 from water). The bacterial count in the positive soil sample was 115 CFU/g. The mean concentrations ± SDs of B. pseudomallei in the positive water and rhizosphere samples were 5.1 ± 5.5 CFU/ml and 80 ± 49 CFU/g, respectively. Six water samples with positive results were collected from a pond and water sources for drinking and daily use. All colonies isolated from the patient shared the same PFGE type (PT) indicating monoclonal infection of ST99. Although the 152 colonies from environmental isolates exhibited 25 PTs, none were identical to the patient’s isolates. PT5 and PT7 were most common genotype among the environmental samples. Conclusions/Significance: Diverse genotypes of B. pseudomallei were prevalent in the environment. However, the patient may have been infected with a low-density genotype. Intervention strategies for preventing B. pseudomallei infection are required. [ABSTRACT FROM AUTHOR]

Background: Leptospirosis is a worldwide zoonotic bacterial disease caused by infection with leptospires. Leptospirosis in humans and livestock is an endemic and epidemic disease in Thailand. Livestock may act as reservoirs for leptospires and source for human infection. Methodology/Principal findings: Data on leptospirosis infection in humans and livestock (Buffaloes, Cattle, and Pigs) species during 2010 to 2015 were analyzed. Serum samples were examined using Microscopic Agglutination Test (MAT) to identify antibodies against Leptospira serovars using a cut-off titer ≥ 1:100. The seroprevalence was 23.7% in humans, 24.8% in buffaloes, 28.1% in cattle, and 11.3% in pigs. Region specific prevalence among humans and livestock was found in a wide range. The most predominant serovars were Shermani, followed by Bratislava, Panama, and Sejroe in human, Shermani, Ranarum, and Tarassovi in buffaloes, and Shermani and Ranarum in cattle and pigs. Equally highest MAT titers against multiple serovars per one sample were found mainly in buffaloes and cattle showing equally titers against Ranarum and Shermani. The correlations of distribution of serovars across Thailand’s regions were found to be similar in pattern for cattle but not for buffaloes. In humans, the serovar distribution in the south differed from other regions. By logistic regression, the results indicated that livestock is more susceptible to infection by serovar Shermani when compared to humans. Conclusions/Significance: This study gives a detailed picture of the predominance of Leptospira serovars in relation to region, humans and typical livestock. The broad spatial distribution of seroprevalence was analyzed across and within species as well as regions in Thailand. Our finding may guide public health policy makers to implement appropriate control measures and help to reduce the impact of leptospirosis in Thailand. [ABSTRACT FROM AUTHOR]

Oral cholera vaccines (OCVs) are relatively new public health interventions, and limited data exist on the potential impact of OCV use on traditional cholera prevention and control measures—safe water, sanitation and hygiene (WaSH). To assess OCV acceptability and knowledge, attitudes, and practices (KAPs) regarding cholera and WaSH, we conducted cross-sectional surveys, 1 month before (baseline) and 3 and 12 months after (first and second follow-up) a preemptive OCV campaign in Maela, a long-standing refugee camp on the Thailand-Burma border. We randomly selected households for the surveys, and administered questionnaires to female heads of households. In total, 271 (77%), 187 (81%), and 199 (85%) households were included in the baseline, first and second follow-up surveys, respectively. Anticipated OCV acceptability was 97% at baseline, and 91% and 85% of household members were reported to have received 1 and 2 OCV doses at first follow-up. Compared with baseline, statistically significant differences (95% Wald confidence interval not overlapping zero) were noted at first and second follow-up among the proportions of respondents who correctly identified two or more means of cholera prevention (62% versus 78% and 80%), reported boiling or treating drinking water (19% versus 44% and 69%), and washing hands with soap (66% versus 77% and 85%); a significant difference was also observed in the proportion of households with soap available at handwashing areas (84% versus 90% and 95%), consistent with reported behaviors. No significant difference was noted in the proportion of households testing positive for Escherichia coli in stored household drinking water at second follow-up (39% versus 49% and 34%). Overall, we observed some positive, and no negative changes in cholera- and WaSH-related KAPs after an OCV campaign in Maela refugee camp. OCV campaigns may provide opportunities to reinforce beneficial WaSH-related KAPs for comprehensive cholera prevention and control. [ABSTRACT FROM AUTHOR]

Background: The rise in dengue fever cases and the absence of dengue vaccines will likely cause governments to consider various types of effective means for controlling the disease. Given strong public interests in potential dengue vaccines, it is essential to understand the private economic benefits of dengue vaccines for accelerated introduction of vaccines into the public sector program and private markets of high-risk countries. Methodology/Principal Findings: A contingent valuation study for a hypothetical dengue vaccine was administered to 400 households in a multi-country setting: Vietnam, Thailand, and Colombia. All respondents received a description of the hypothetical dengue vaccine scenarios of 70% or 95% effectiveness for 10 or 30 years with a three dose series. Five price points were determined after pilot tests in order to reflect different local situations such as household income levels and general perceptions towards dengue fever. We adopted either Poisson or negative binomial regression models to calculate average willingness-to-pay (WTP), as well as median WTP. We found that there is a significant demand for dengue vaccines. The parametric median WTP is $26.4 ($8.8 per dose) in Vietnam, $70.3 ($23.4 per dose) in Thailand, and $23 ($7.7 per dose) in Colombia. Our study also suggests that respondents place more value on vaccinating young children than school age children and adults. Conclusions/Significance: Knowing that dengue vaccines are not yet available, our study provides critical information to both public and private sectors. The study results can be used to ensure broad coverage with an affordable price and incorporated into cost benefit analyses, which can inform prioritization of alternative health interventions at the national level. [ABSTRACT FROM AUTHOR]

Melioidosis is an often fatal infectious disease affecting humans and animals in tropical regions and is caused by the saprophytic environmental bacterium Burkholderia pseudomallei. Domestic gardens are not only a common source of exposure to soil and thus to B. pseudomallei, but they also have been found to contain more B. pseudomallei than other environments. In this study we addressed whether anthropogenic manipulations common to gardens such as irrigation or fertilizers change the occurrence of B. pseudomallei. We conducted a soil microcosm experiment with a range of fertilizers and soil types as well as a longitudinal interventional study over three years on an experimental fertilized field site in an area naturally positive for B. pseudomallei. Irrigation was the only consistent treatment to increase B. pseudomallei occurrence over time. The effects of fertilizers upon these bacteria depended on soil texture, physicochemical soil properties and biotic factors. Nitrates and urea increased B. pseudomallei load in sand while phosphates had a positive effect in clay. The high buffering and cation exchange capacities of organic material found in a commercial potting mix led to a marked increase in soil salinity with no survival of B. pseudomallei after four weeks in the potting mix sampled. Imported grasses were also associated with B. pseudomallei occurrence in a multivariate model. With increasing population density in endemic areas these findings inform the identification of areas in the anthropogenic environment with increased risk of exposure to B. pseudomallei. Author Summary: Melioidosis cases are on the rise in endemic areas of northern Australia and Thailand. This potentially severe infectious disease affecting humans and animals in the tropical belt is caused by the gram negative bacterium Burkholderia pseudomallei. Domestic gardens are a common point of exposure to these environmental bacteria and B. pseudomallei are more prevalent in the dry season in gardens when compared to other areas. This is why we analysed whether common gardening practices such as regular watering (irrigation) or soil fertilizing change the occurrence of B. pseudomallei. We conducted a soil microcosm experiment with a range of fertilizers and soil types as well as a longitudinal interventional study over three years on an experimental fertilized field site in an area naturally positive for B. pseudomallei. Irrigation was the only consistent treatment to increase B. pseudomallei occurrence over time. The effects of fertilizers upon these bacteria depended on soil texture, physicochemical properties such as pH or salinity and vegetation. B. pseudomallei occurrence was also associated with imported grasses. With increasing populations in endemic areas, these findings inform the identification of areas in the anthropogenic environment with increased risk of exposure to B. pseudomallei. [ABSTRACT FROM AUTHOR]

The dynamics of Plasmodium vivax infection is characterized by reactivation of hypnozoites at varying time intervals. The relative contribution of new P. vivax infection and reactivation of dormant liver stage hypnozoites to initiation of blood stage infection is unclear. In this study, we investigate the contribution of new inoculations of P. vivax sporozoites to primary infection versus reactivation of hypnozoites by modeling the dynamics of P. vivax infection in Thailand in patients receiving treatment for either blood stage infection alone (chloroquine), or the blood and liver stages of infection (chloroquine + primaquine). In addition, we also analysed rates of infection in a study in Papua New Guinea (PNG) where patients were treated with either artesunate, or artesunate + primaquine. Our results show that up to 96% of the P. vivax infection is due to hypnozoite reactivation in individuals living in endemic areas in Thailand. Similar analysis revealed the around 70% of infections in the PNG cohort were due to hypnozoite reactivation. We show how the age of the cohort, primaquine drug failure, and seasonality may affect estimates of the ratio of primary P. vivax infection to hypnozoite reactivation. Modeling of P. vivax primary infection and hypnozoite reactivation provides important insights into infection dynamics, and suggests that 90–96% of blood stage infections arise from hypnozoite reactivation. Major differences in infection kinetics between Thailand and PNG suggest the likelihood of drug failure in PNG. Author Summary: Plasmodium vivax is one of two major parasite species causing human disease. This parasite can lie dormant in the liver as a hypnozoite, before later reactivating to cause blood-stage infection. Treatment to eliminate the dormant hypnozoite stage relies mostly on a single drug—primaquine. Understanding the rate of primary infection versus hypnozoite reactivation is important to understanding primaquine efficacy and drug resistance, as well as the development of new drugs targeting hypnozoites. Here we use mathematical modeling to analyse data from two clinical cohorts and show that up to 96% of infections may be caused by hypnozoite reactivation. We also use modeling to understand the impact of drug resistance, seasonal infection and subject age. [ABSTRACT FROM AUTHOR]

Snake envenomation has been estimated to affect 1.8 million people annually with about 94,000 deaths mostly in poor tropical countries. Specific antivenoms are the only rational and effective therapy for these cases. Efforts are being made to produce effective, affordable and sufficient antivenoms for these victims. The immunization process, which has rarely been described in detail, is one step that needs to be rigorously studied and improved especially with regard to the production of polyspecific antisera. The polyspecific nature of therapeutic antivenom could obviate the need to identify the culprit snake species. The aim of this study was to produce potent polyspecific antisera against 3 medically important vipers of Thailand and its neighboring countries, namely Cryptelytrops albolabris "White lipped pit viper" (CA), Calleoselasma rhodostoma "Malayan pit viper" (CR), and Daboia siamensis "Russell's viper" (DS). Four horses were immunized with a mixture of the 3 viper venoms using the 'low dose, low volume multi-site' immunization protocol. The antisera showed rapid rise in ELISA titers against the 3 venoms and reached plateau at about the 8th week post-immunization. The in vivo neutralization potency (P) of the antisera against CA, CR and DS venoms was 10.40, 2.42 and 0.76 mg/ml, respectively and was much higher than the minimal potency limits set by Queen Soavabha Memorial Institute (QSMI). The corresponding potency values for the QSMI monospecific antisera against CA, CR and DS venoms were 7.28, 3.12 and 1.50 mg/ml, respectively. The polyspecific antisera also effectively neutralized the procoagulant, hemorrhagic, necrotic and nephrotoxic activities of the viper venoms. This effective immunization protocol should be useful in the production of potent polyspecific antisera against snake venoms, and equine antisera against tetanus, diphtheria or rabies. Author Summary: Snake envenomation is a serious medical problem in various tropical developing countries. Though antivenoms are the main and rational treatment, they are often of low potency, expensive and/or not available. Moreover, most antivenoms are specific for the treatment of one single snake species and thus necessitate catching the culprit snake so the correct antivenom can be administered. Studies currently are being conducted with the aim of eliminating these shortcomings and to produce polyspecific antivenom capable of neutralizing multiple snake venoms. Production of snake antivenoms involves immunization of an animal, usually horses, with snake venom(s). The serum or plasma is then fractionated to yield antivenom antibodies. While the fractionation processes have been well established, the immunization protocol for the preparation of high potency antisera has rarely been reported in detail. This report describes an effective immunization protocol for the production of potent polyspecific horse antisera against all 3 medically important viper venoms of Thailand (Russell's viper, Malayan pit viper and green pit viper). The antivenom prepared from the antisera should be useful for the treatment of these viper bites. The immunization protocol should be useful in the production of other potent polyspecific antisera as well as equine antisera against other diseases. [ABSTRACT FROM AUTHOR]

Background: Dengue control programs commonly employ reactive insecticide spraying around houses of reported cases, with the assumption that most dengue virus (DENV) transmission occurs in the home. Focal household transmission has been demonstrated in rural settings, but it is unclear whether this holds true in dense and mobile urban populations. We conducted a prospective study of dengue clustering around households in highly urban Ho Chi Minh City, Vietnam. Methods: We enrolled 71 index cases with suspected dengue (subsequently classified as 52 dengue cases and 19 non-dengue controls); each initiated the enrollment of a cluster of 25–35 household members and neighbors who were followed up over 14 days. Incident DENV infections in cluster participants were identified by RT-PCR, NS1-ELISA, and/or DENV-IgM/-IgG seroconversion, and recent infections by DENV-IgM positivity at baseline. Principal Findings/Conclusions: There was no excess risk of DENV infection within dengue case clusters during the two-week follow-up, compared to control clusters, but the prevalence of recent DENV infection at baseline was two-fold higher in case clusters than controls (OR 2.3, 95%CI 1.0–5.1, p = 0.05). Prevalence of DENV infection in Aedes aegypti was similar in case and control houses, and low overall (1%). Our findings are broadly consistent with household clustering of dengue risk, but indicate that any clustering is at a short temporal scale rather than sustained chains of localized transmission. This suggests that reactive perifocal insecticide spraying may have a limited impact in this setting. Author Summary: Dengue is a viral disease transmitted between humans by Aedes aegypti mosquitoes. It is an increasing health problem in tropical regions, with no vaccine or drug available. The main approach used for dengue prevention and control is reducing mosquito numbers, which is commonly done by spraying insecticide around the houses of reported dengue cases and their neighbors. This assumes that mosquito populations around the home contribute to onwards transmission of dengue to other people living near the identified case, and that by reducing these populations further infections can be prevented. A previous study in rural villages in Thailand showed that dengue infections did cluster near the house of a case, but it is not clear whether this would also be the case in a densely-populated urban setting like Ho Chi Minh City. In this study, we show that people living close to a dengue case in HCMC were in fact no more likely to be infected with dengue over a two week period than controls who did not live near a dengue case, although they were two times more likely to have evidence of a recent dengue infection at enrollment. This suggests that dengue risk might cluster around households in this setting, but that responsive insecticide spraying around the homes of reported cases is unlikely to prevent many additional infections. [ABSTRACT FROM AUTHOR]

Since 1996, there have been several case reports of autochthonous visceral leishmaniasis in Thailand. Here we report a case in a 52-year-old Thai male from northern Thailand, who presented with subacute fever, huge splenomegaly and pancytopenia. Bone marrow aspiration revealed numerous amastigotes within macrophages. Isolation of Leishmania LSCM1 into culture and DNA sequence analysis (ribosomal RNA ITS-1 and large subunit of RNA polymerase II) revealed the parasites to be members of the Leishmania enriettii complex, and apparently identical to L. martiniquensis previously reported from the Caribbean island of Martinique. This is the first report of visceral leishmaniasis caused by L. martiniquensis from the region. Moreover, the majority of parasites previously identified as "L. siamensis" also appear to be L. martiniquensis. Author Summary: Visceral leishmaniasis is a potentially life threatening parasitic disease that is found in many tropical and subtropical regions of the world. We report a case of visceral leishmaniasis in northern Thailand caused by a recently described new species of Leishmania parasite, Leishmania martiniquensis. By comparing DNA sequences from this parasite with those of other isolates, we show that this parasite is more widely spread than previously thought, with several other examples in Thailand, and possibly from Europe and the USA, as well as the original location in the Caribbean island of Martinique. This parasite appears to be an emerging pathogen, which, because it causes a potentially fatal disease, needs to be investigated in detail so that its transmission can be understood and control measures can be developed. [ABSTRACT FROM AUTHOR]

Background: Malaria control efforts have a significant impact on the epidemiology and parasite population dynamics. In countries aiming for malaria elimination, malaria transmission may be restricted to limited transmission hot spots, where parasite populations may be isolated from each other and experience different selection forces. Here we aim to examine the Plasmodium vivax population divergence in geographically isolated transmission zones in Thailand. Methodology: We employed the P. vivax merozoite surface protein 3β (PvMSP3β) as a molecular marker for characterizing P. vivax populations based on the extensive diversity of this gene in Southeast Asian parasite populations. To examine two parasite populations with different transmission levels in Thailand, we obtained 45 P. vivax isolates from Tak Province, northwestern Thailand, where the annual parasite incidence (API) was more than 2%, and 28 isolates from Yala and Narathiwat Provinces, southern Thailand, where the API was less than 0.02%. We sequenced the PvMSP3β gene and examined its genetic diversity and molecular evolution between the parasite populations. Principal Findings: Of 58 isolates containing single PvMSP3β alleles, 31 sequence types were identified. The overall haplotype diversity was 0.77±0.06 and nucleotide diversity 0.0877±0.0054. The northwestern vivax malaria population exhibited extensive haplotype diversity (HD) of PvMSP3β (HD = 1.0). In contrast, the southern parasite population displayed a single PvMSP3β allele (HD = 0), suggesting a clonal population expansion. This result revealed that the extent of allelic diversity in P. vivax populations in Thailand varies among endemic areas. Conclusion: Malaria parasite populations in a given region may vary significantly in genetic diversity, which may be the result of control and influenced by the magnitude of malaria transmission intensity. This is an issue that should be taken into account for the implementation of P. vivax control measures such as drug policy and vaccine development. Author Summary: With intensified malaria control in endemic countries, there have been dramatic changes of malaria epidemiology. One of such changes is the increased proportion of Plasmodium vivax malaria, a demonstration of resilience of this parasite to control efforts. In Thailand, malaria has been largely eliminated from the central plain, and transmission is concentrated in isolated international border regions. This study aimed to examine whether the changing malaria epidemiology was reflected in the population dynamics and genetic diversity of the isolated parasite populations. We collected parasite samples from two regions in Thailand with drastically different endemicity settings and used a polymorphic genetic marker (Plasmodium vivax merozoite surface protein 3β – pvmsp3β) as an indicator of genetic diversity of the populations. Analysis of the pvmsp3β sequences revealed high genetic diversity of parasites from western Thailand, and suggested the suitability of this gene as a molecular marker to infer parasite genetic diversity. Comparing the pvmsp3β sequences, we further discovered extreme divergence in genetic diversity between the southern and northwestern Thai P. vivax populations. Our study offers important insights into malaria epidemiology and provides the needed knowledge for designing novel control tools in the malaria elimination campaigns. [ABSTRACT FROM AUTHOR]

Background: Dengue is endemic to the rural province of Kamphaeng Phet, Northern Thailand. A decade of prospective cohort studies has provided important insights into the dengue viruses and their generated disease. However, as elsewhere, spatial dynamics of the pathogen remain poorly understood. In particular, the spatial scale of transmission and the scale of clustering are poorly characterized. This information is critical for effective deployment of spatially targeted interventions and for understanding the mechanisms that drive the dispersal of the virus. Methodology/Principal Findings: We geocoded the home locations of 4,768 confirmed dengue cases admitted to the main hospital in Kamphaeng Phet province between 1994 and 2008. We used the phi clustering statistic to characterize short-term spatial dependence between cases. Further, to see if clustering of cases led to similar temporal patterns of disease across villages, we calculated the correlation in the long-term epidemic curves between communities. We found that cases were 2.9 times (95% confidence interval 2.7–3.2) more likely to live in the same village and be infected within the same month than expected given the underlying spatial and temporal distribution of cases. This fell to 1.4 times (1.2–1.7) for individuals living in villages 1 km apart. Significant clustering was observed up to 5 km. We found a steadily decreasing trend in the correlation in epidemics curves by distance: communities separated by up to 5 km had a mean correlation of 0.28 falling to 0.16 for communities separated between 20 km and 25 km. A potential explanation for these patterns is a role for human movement in spreading the pathogen between communities. Gravity style models, which attempt to capture population movement, outperformed competing models in describing the observed correlations. Conclusions: There exists significant short-term clustering of cases within individual villages. Effective spatially and temporally targeted interventions deployed within villages may target ongoing transmission and reduce infection risk. Author Summary: Transmission of dengue virus has long been studied in Kamphaeng Phet, Northern Thailand, but how cases are related in time and space is still unclear, as is the role of human movement in generating these patterns. Because of these knowledge gaps, public health officials cannot make educated decisions on how to target vector control interventions and mechanisms of virus dispersal are not known. We mapped the homes of dengue cases admitted to the main hospital in the province capital from 1994–2008 and quantified the spatial correlation between them. We found an almost three times greater chance that cases from the same month came from the same village than expected, given the overall distribution of cases. Some clustering was also observed between cases in neighboring villages with the overall epidemics experienced by neighboring communities also more correlated than epidemics in villages farther apart. The short-term clustering observed within individual villages implies that effective spatially targeted interventions deployed within villages may reduce infection risk. As the distance between neighboring communities exceeds the typical flight range of the dengue vector, these findings also suggest a potential role for human movement in driving the wider spread of the virus. [ABSTRACT FROM AUTHOR]

Background: Mosquito biting frequency and how bites are distributed among different people can have significant epidemiologic effects. An improved understanding of mosquito vector-human interactions would refine knowledge of the entomological processes supporting pathogen transmission and could reveal targets for minimizing risk and breaking pathogen transmission cycles. Methodology and principal findings: We used human DNA blood meal profiling of the dengue virus (DENV) vector, Aedes aegypti, to quantify its contact with human hosts and to infer epidemiologic implications of its blood feeding behavior. We determined the number of different people bitten, biting frequency by host age, size, mosquito age, and the number of times each person was bitten. Of 3,677 engorged mosquitoes collected and 1,186 complete DNA profiles, only 420 meals matched people from the study area, indicating that Ae. aegypti feed on people moving transiently through communities to conduct daily business. 10–13% of engorged mosquitoes fed on more than one person. No biting rate differences were detected between high- and low-dengue transmission seasons. We estimate that 43–46% of engorged mosquitoes bit more than one person within each gonotrophic cycle. Most multiple meals were from residents of the mosquito collection house or neighbors. People ≤25 years old were bitten less often than older people. Some hosts were fed on frequently, with three hosts bitten nine times. Interaction networks for mosquitoes and humans revealed biologically significant blood feeding hotspots, including community marketplaces. Conclusion and significance: High multiple-feeding rates and feeding on community visitors are likely important features in the efficient transmission and rapid spread of DENV. These results help explain why reducing vector populations alone is difficult for dengue prevention and support the argument for additional studies of mosquito feeding behavior, which when integrated with a greater understanding of human behavior will refine estimates of risk and strategies for dengue control. Author Summary: Dengue, a potentially lethal infection impacting hundreds of millions of human lives annually, is caused by viruses transmitted during mosquito blood feeding. With no vaccine or treatment commercially available, understanding the underlying factors linked to virus exposure is critical for developing more effective dengue interventions. We conducted a study in an endemic region of Thailand where transmission is high and children are expected to be the non-immune, amplifying portion of the host population. We examined Ae. aegypti feeding patterns and risk by matching human DNA profiles in blood-fed mosquitoes to study area residents. A small number of meals matched people from the study area, suggesting that mosquitoes feed on people moving transiently through communities. People under 25 years of age were bitten less frequently than older people. We constructed network models to explore the presence of mosquito feeding "hotspots" and detected a local market "hotspot" in one study village during the high dengue transmission season. Our results provide new details on dengue vector feeding patterns and highlight the need to conduct integrated studies of vector feeding and human behavior, and virus transmission patterns in order to better understand the dengue transmission efficiency and spread. [ABSTRACT FROM AUTHOR]