Bridging Biodiversity and Health: The Global Biodiversity Information Facility's initiative on open data on vectors of human diseases.

There is an increased awareness of the importance of data publication, data sharing, and open science to support research, monitoring and control of vector-borne disease (VBD). Here we describe the efforts of the Global Biodiversity Information Facility (GBIF) as well as the World Health Special Programme on Research and Training in Diseases of Poverty (TDR) to promote publication of data related to vectors of diseases. In 2020, a GBIF task group of experts was formed to provide advice and support efforts aimed at enhancing the coverage and accessibility of data on vectors of human diseases within GBIF. Various strategies, such as organizing training courses and publishing data papers, were used to increase this content. This editorial introduces the outcome of a second call for data papers partnered by the TDR, GBIF and GigaScience Press in the journal GigaByte. Biodiversity and infectious diseases are linked in complex ways. These links can involve changes from the microorganism level to that of the habitat, and there are many ways in which these factors interact to affect human health. One way to tackle disease control and possibly elimination, is to provide stakeholders with access to a wide range of data shared under the FAIR principles, so it is possible to support early detection, analyses and evaluation, and to promote policy improvements and/or development.

Effects of mosquito age and batch size on knockdown and mortality of laboratory-reared Anopheles dirus, Anopheles minimus, and wild-caught Anopheles harrisoni (Diptera: Culicidae) exposed to transfluthrin using WHO tube and CDC bottle bioassays.

WHO tube and CDC bottle bioassays are currently available for insecticide resistance monitoring and malaria transmission research. Multiple parameters including mosquito density, age, and nutritional status may affect the readout in these bioassays' tests. This study aims to assess the effects of experimental factors on knockdown and mortality measurements in dominant malaria vectors in Thailand following exposure to sublethal and lethal doses of transfluthrin. The effects of (i) 3 different mosquito batch sizes (5, 10, and 20 individuals) and (ii) 2 age groups (3-5 and 20-23 days old) on outcomes measured using the WHO tube (14.7 µg/cm2) and CDC bottle bioassay discriminating concentration (0.006 µg/cm2) against 2 laboratory strains: Anopheles dirus Peyton & Harrison and Anopheles minimus Theobald (species A) and wild-caught Anopheles harrisoni Harbach & Manguin (species C). Our results showed higher knockdown at 1-h exposure using WHO tube and CDC bottle bioassays containing 20 individuals compared to batches containing 10 and 5 individuals. Older mosquitoes showed greater susceptibility than younger test population, especially for An. mininus. Our study supports WHO recommendations for using 3- to 5-day-old mosquitoes. It also validates Praulin et al. (2022) proposal to divide the cohort into smaller batches with more test replicates when it is not practicable to test 25 mosquitoes per replicate.

Evolution and spread of Plasmodium falciparum mutations associated with resistance to sulfadoxine-pyrimethamine in central Africa: a cross-sectional study.

BACKGROUND: Efficacy of sulfadoxine-pyrimethamine, the malaria chemoprophylaxis used in pregnant women, and in children when combined with amodiaquine, is threatened by the accumulation of mutations in the Plasmodium falciparum dihydropteroate synthase (pfdhps) and dihydrofolate reductase (pfdhfr) genes. Data on the prevalence of resistant alleles in central Africa and the new pfdhps I431V mutation, particularly associated with other mutations to form the pfdhps vagKgs allele, are scarce. We explored the frequency and geographical distribution of pfdhps and pfdhfr mutations in central Africa in 2014-18, and assessed the evolutionary origin of the vagKgs allele. METHODS: Samples were collected at 18 health-care centres in seven countries (Angola, Cameroon, Central African Republic, Democratic Republic of the Congo, Gabon, Nigeria, and Republic of the Congo) from patients who showed possible symptoms of malaria between March 1, 2014, and Oct 31, 2018. Samples that were positive for P falciparum were transported to a laboratory in Toulouse, France, and genotyped. The frequency of pfdhfr and pfdhps mutations was studied in 1749 samples. Microsatellites in pfdhps flanking regions and whole-genome analysis compared with parasite genomes from the data-sharing network MalariaGEN were performed on samples carrying the vagKgs allele. FINDINGS: Mapping of the prevalence of single nucleotide polymorphisms and corresponding alleles of pfdhfr and pfdhps showed a substantial spread of alleles associated with sulfadoxine-pyrimethamine resistance in central Africa during the 2014-18 period, especially an increase going west to east in pfdhps alleles carrying the K540E and A581G mutations. A high prevalence of the pfdhps I431V mutation was observed in Cameroon (exceeding 50% in the northern region) and Nigeria. Genomic analysis showed a recent African emergence and a clonal expansion of the most frequent pfdhps vagKgs allele. INTERPRETATION: Reduced sulfadoxine-pyrimethamine efficacy due to increased resistance is a worrying situation, especially because the malaria transmission level is high in central Africa. Although the resistance phenotype remains to be confirmed, the emergence and spread of the vagKgs allele in west and central Africa could challenge the use of sulfadoxine-pyrimethamine. FUNDING: Toulouse Institute for Infectious and Inflammatory Diseases.

Distribution of mosquitoes (Diptera: Culicidae) in Thailand: a dataset.

Mosquitoes play a crucial role as primary vectors for various infectious diseases in Thailand. Therefore, accurate distribution information is vital for effectively combating and better controlling mosquito-borne diseases. Here, we present a curated dataset of the mosquito distribution in Thailand comprising 12,278 records of at least 117 mosquito species (Diptera: Culicidae). The main genera included in the dataset are Aedes, Anopheles, Armigeres, Culex, and Mansonia. From 2007 to 2023, data were collected through routine mosquito surveillance and research projects from 1,725 locations across 66 (out of 77) Thai provinces. The majority of the data were extracted from a Thai database of the Thailand Malaria Elimination Program. To facilitate broader access to mosquito-related data and support further exploration of the Thai mosquito fauna, the data were translated into English. Our dataset has been published in the Global Biodiversity Information Facility, making it available for researchers worldwide.

Synergistic Repellent and Irritant Effects of a Mixture of ß-Caryophyllene Oxide and Vetiver Oil against Mosquito Vectors.

Repellents play a major role in reducing the risk of mosquito-borne diseases by preventing mosquito bites. The present study evaluated the mosquito-repellent activity of ß-caryophyllene oxide 1% (BCO), vetiver oil 2.5% (VO), and their binary mixtures (BCO + VO (1:1), BCO + VO (2:1), BCO + VO (1:2)) against four laboratory-colonized mosquito species, Aedes aegypti (L.), Aedes albopictus (Skuse), Anopheles minimus Theobald, and Culex quinquefasciatus Say, using an excito-repellency assay system. In general, the compound mixtures produced a much stronger response in the mosquitoes than single compounds, regardless of the test conditions or species. The greatest synergetic effect was achieved with the combination of BCO + VO (1:2) in both contact and noncontact trials with An. minimus (74.07-78.18%) and Cx. quinquefasciatus (55.36-83.64%). Knockdown responses to the binary mixture of BCO + VO were observed for Ae. albopictus, An. minimus, and Cx. quinquefasciatus, in the range of 18.18-33.33%. The synergistic repellent activity of BCO and VO used in this study may support increased opportunities to develop safer alternatives to synthetic repellents for personal protection against mosquitoes.

Biodiversity data supports research on human infectious diseases: Global trends, challenges, and opportunities.

The unprecedented generation of large volumes of biodiversity data is consistently contributing to a wide range of disciplines, including disease ecology. Emerging infectious diseases are usually zoonoses caused by multi-host pathogens. Therefore, their understanding may require the access to biodiversity data related to the ecology and the occurrence of the species involved. Nevertheless, despite several data-mobilization initiatives, the usage of biodiversity data for research into disease dynamics has not yet been fully leveraged. To explore current contribution, trends, and to identify limitations, we characterized biodiversity data usage in scientific publications related to human health, contrasting patterns of studies citing the Global Biodiversity Information Facility (GBIF) with those obtaining data from other sources. We found that the studies mainly obtained data from scientific literature and other not aggregated or standardized sources. Most of the studies explored pathogen species and, particularly those with GBIF-mediated data, tended to explore and reuse data of multiple species (>2). Data sources varied according to the taxa and epidemiological roles of the species involved. Biodiversity data repositories were mainly used for species related to hosts, reservoirs, and vectors, and barely used as a source of pathogens data, which was usually obtained from human and animal-health related institutions. While both GBIF- and not GBIF-mediated data studies explored similar diseases and topics, they presented discipline biases and different analytical approaches. Research on emerging infectious diseases may require the access to geographical and ecological data of multiple species. The One Health challenge requires interdisciplinary collaboration and data sharing, which is facilitated by aggregated repositories and platforms. The contribution of biodiversity data to understand infectious disease dynamics should be acknowledged, strengthened, and promoted.

Scientific achievements and reflections after 20 years of vector biology and control research at the Pu Teuy mosquito field research station, Thailand.

Additional vector control tools are needed to supplement current strategies to achieve malaria elimination and control of Aedes-borne diseases in many settings in Thailand and the Greater Mekong Sub-region. Within the next decade, the vector control community, Kasetsart University (KU), and the Ministry of Higher Education, Science, Research and Innovation must take full advantage of these tools that combine different active ingredients with different modes of action. Pu Teuy Mosquito Field Research Station (MFRS), Department of Entomology, Faculty of Agriculture, Kasetsart University (KU), Thailand was established in 2001 and has grown into a leading facility for performing high-quality vector biology and control studies and evaluation of public health insecticides that are operationally relevant. Several onsite mosquito research platforms have been established including experimental huts, a 40-m long semi-field screening enclosure, mosquito insectary, field-laboratory, and living quarters for students and researchers. Field research and assessments ranged from 'basic' investigations on mosquito biology, taxonomy and genetics to more 'applied' studies on responses of mosquitoes to insecticides including repellency, behavioural avoidance and toxicity. In the course of two decades, 51 peer-reviewed articles have been published, and 7 masters and 16 doctoral degrees in Entomology have been awarded to national and international students. Continued support of key national stakeholders will sustain MFRS as a Greater Mekong Subregion centre of excellence and a resource for both insecticide trials and entomological research.

Behavioral avoidance and biological safety of vetiver oil and its constituents against Aedes aegypti (L.), Aedes albopictus (Skuse) and Culex quinquefasciatus Say.

Numerous plant-based repellents are widely used for personal protection against host-seeking mosquitoes. Vitiveria zizanioides (L.) Nash essential oil and its constituents have demonstrated various mosquito repellent activities. In this study, three chemical actions of vetiver oil and five constituents (terpinen-4-ol, α-terpineol, valencene, vetiverol and vetivone) were characterized against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus by using the high-throughput screening assay system (HITSS). Significant contact escape responses in Ae. aegypti and Ae. albopictus to all test compounds at concentrations between 2.5 and 5% were observed. Spatial repellency responses were also observed in some tested mosquito populations depending upon concentrations. The most significant toxic response on mosquitoes was found at the highest concentration, except for vetivone which had no toxic effect on Ae. aegypti and Ae. albopictus. Results on phototoxic and genotoxic hazard revealed that vetiver oil and their constituents showed no phototoxic potential or any significant genotoxic response. In conclusion, vetiver oil and two constituents, valencene and vetiverol, are potentials as active ingredients for mosquito repellency and present no toxicity.

Prevalence and spatial distribution characteristics of human echinococcosis in China.

BACKGROUND: Echinococcosis is a zoonotic parasitic disease caused by larval stages of cestodes belonging to the genus Echinococcus. The infection affects people's health and safety as well as agropastoral sector. In China, human echinococcosis is a major public health burden, especially in western China. Echinococcosis affects people health as well as agricultural and pastoral economy. Therefore, it is important to understand the prevalence status and spatial distribution of human echinococcosis in order to advance our knowledge of basic information for prevention and control measures reinforcement. METHODS: Report data on echinococcosis were collected in 370 counties in China in 2018 and were used to assess prevalence and spatial distribution. SPSS 21.0 was used to obtain the prevalence rate for CE and AE. For statistical analyses and mapping, all data were processed using SPSS 21.0 and ArcGIS 10.4, respectively. Chi-square test and Exact probability method were used to assess spatial autocorrelation and spatial clustering. RESULTS: A total of 47,278 cases of echinococcosis were recorded in 2018 in 370 endemic counties in China. The prevalence rate of human echinococcosis was 10.57 per 10,000. Analysis of the disease prevalence showed obvious spatial positive autocorrelation in globle spatial autocorrelation with two aggregation modes in local spatial autocorrelation, namely high-high and low-high aggregation areas. The high-high gathering areas were mainly concentrated in northern Tibet, western Qinghai, and Ganzi in the Tibetan Autonomous Region and in Sichuan. The low-high clusters were concentrated in Gamba, Kangma and Yadong counties of Tibet. In addition, spatial scanning analysis revealed two spatial clusters. One type of spatial clusters included 71 counties in Tibet Autonomous Region, 22 counties in Qinghai, 11 counties in Sichuan, three counties in Xinjiang Uygur Autonomous Region, two counties in Yunnan, and one county in Gansu. In the second category, six types of spatial clusters were observed in the counties of Xinjiang Uygur Autonomous Region, and the Qinghai, Gansu, and Sichuan Provinces. CONCLUSION: This study showed a serious prevalence of human echinococcosis with obvious spatial aggregation of the disease prevalence in China. The Qinghai-Tibet Plateau is the "hot spot" area of human echinococcosis in China. Findings from this study indicate that there is an urgent need of joint strategies to strengthen efforts for the prevention and control of echinococcosis in China, especially in the Qinghai-Tibet Plateau.

Homogeneity and Possible Replacement of Populations of the Dengue Vectors Aedes aegypti and Aedes albopictus in Indonesia.

Currently, Aedes aegypti, the principal vector of dengue virus in Indonesia, has spread throughout the archipelago. Aedes albopictus is also present. Invasion and high adaptability of the Aedes mosquitoes to all of these areas are closely related to their ecology and biology. Between June 2016 and July 2017, larval and adult mosquito collections were conducted in 43 locations in 25 provinces of Indonesia using standardized sampling methods for dengue vector surveillance. The samples collected were analyzed for polymorphism and phylogenetic relationship using the mitochondrial cox1 gene and the nuclear ribosomal internal transcribed spacer 2 (ITS2). Almost all Ae. aegypti samples collected in this study (89%) belonged to the same haplotype. A similar situation is observed with the nuclear ITS2 marker. Populations of Ae. aegypti characterized few years ago were genetically different. A closely related observation was made with Aedes albopictus for which the current populations are different from those described earlier. Ae. aegypti populations were found to be highly homogenous all over Indonesia with all samples belonging to the same maternal lineage. Although difficult to demonstrate formally, there is a possibility of population replacement. Although to a lower extent, a similar conclusion was reached with Ae. albopictus.

Defeating malaria in the North-East region: the forerunner for malaria elimination in India.

India is a malaria endemic country which is targeting malaria elimination by 2027. Transmission intensities are low-to-moderate depending on the region supported by multiple disease vectors. Among these, comparatively North-East India contributes to high proportions of malaria cases annually, the majority of which are due to Plasmodium falciparum (90%). Anopheles minimus and An. baimaii (sibling species in the An. dirus complex) are widely prevalent and incriminated as vectors of malaria. Number of intervention tools were field-evaluated beginning 1988 to date against disease vectors and causative parasites to contain the spread of malaria. These included (i) insecticide-treated netting materials (ITNs) for vector control, (ii) rapid diagnostic tests (RDTs) for in situ diagnosis, and (iii) therapeutic efficacy of artemisinin-based combination therapies (ACTs) for improved drug-policy; all of which were incorporated in healthcare services resulting in substantial disease transmission reduction. Populations of both An. minimus and An. baimaii were observed depleting, instead An. culicifacies s.l. recorded to be fast invading degraded forests and assessed to be resistant to multiple insecticides. Of the two prevalent Plasmodium species, while P. vivax continued to be susceptible to chloroquine therapy, P. falciparum had emerged resistant to most available antimalarial drugs except ACTs over space and time and spreading to peninsular India threatening elimination efforts. Disease transmission trends were observed to be declining for which the state of Assam has made huge strides reporting steady fall in cases each passing year vis-à-vis Meghalaya, Mizoram and Tripura (all sharing international border with Bangladesh), in which malaria transmission remained uninterrupted. Consequently, control of malaria in the North-East region of India is of immediate importance and needs prioritization for intensified disease surveillance and control interventions coupled with improved access to healthcare services mitigating risk of disease outbreaks and spread of drug-resistant malaria helping realize the goal of malaria elimination in the country.

Assessment of Mosquito Collection Methods for Dengue Surveillance.

Several methods exist to collect and assess the abundance of dengue vector mosquitoes, i.e., morning adult collection, pupal collection, ovitraps, human landing, and larval collection. Several of these methods are officially implemented to monitor mosquito density and make decisions on treatments for dengue control. This monitoring is also constrained by the need to conduct this assessment on a "one point/one day" process, meaning that once the threshold of 100 households is reached, the assessment is made, and the collectors teams move to another place, thus preventing the use of long-term sampling methods. This diversity of methods might be a source of variability and lack of statistical significance. There is also a lack of published data regarding the efficacy of these methods. Furthermore, the Stegomyia indices are shown to be not reliable for assessing the risk of dengue outbreaks. A mosquito survey was, thus, conducted in 39 locations corresponding to 15 dengue endemic provinces in Indonesia by using the different adult and larval collection methods recommended nationwide. A total of 44,675 mosquitoes were collected. The single larva method was the most efficient. Out of a total of 89 dengue-positive pools, the most frequently encountered virus was DENV2, which made up half of the positive samples, followed by DENV3 and DENV1, respectively. Factor analysis of mixed data showed that no correlation could be found between any methods and the presence of dengue virus in mosquitoes. Moreover, no correlation could be found between any methods and the incidence of dengue. There was no consistency in the efficacy of a given method from one site to another. There was no correlation between any of the parameters considered, i.e., method, incidence of dengue, location, and the presence of dengue virus in mosquitoes.

Fast Expansion of the Asian-Pacific Genotype of the Chikungunya Virus in Indonesia.

Chikungunya is repeatedly affecting Indonesia through successive outbreaks. The Asian genotype has been present in Asia since the late 1950s while the ECSA-IOL (East/Central/South Africa - Indian Ocean Lineage) genotype invaded Asia in 2005. In order to determine the extension of the circulation of the chikungunya virus (CHIKV) in Indonesia, mosquitoes were collected in 28 different sites from 12 Indonesian provinces in 2016-2017. The E1 subunit of the CHIKV envelope gene was sequenced while mosquitoes were genotyped using the mitochondrial cox1 (cytochrome C oxidase subunit 1) gene to determine whether a specific population was involved in the vectoring of CHIKV. A total of 37 CHIKV samples were found in 28 Aedes aegypti, 8 Aedes albopictus and 1 Aedes butleri out of 15,362 samples collected and tested. These viruses, like all Indonesian CHIKV since 2000, belonged to a genotype we propose to call the Asian-Pacific genotype. It also comprises the Yap isolates and viruses having emerged in Polynesia, the Caribbean and South America. They differ from the CHIKV of the Asian genotype found earlier in Indonesia indicating a replacement. These results raise the question of the mechanisms behind this fast and massive replacement.

Trophic Behavior and Species Diversity of the Anopheles barbirostris Complex (Diptera: Culicidae) in Thailand.

Species of the Anopheles barbirostris complex (Myzorhynchus Series of the subgenus Anopheles) are potential vectors of malaria and filariasis parasites. Owing to the lack of reliable identification methods, the biting activity and host preference of the species within this complex have not been previous described. In this study, the trophic behavior and host preferences of the species in the complex were determined in Thailand, and a map of their geographical distributions constructed. Adult female mosquitoes in Chanthaburi and Prachuap Khiri Khan provinces were collected using human landings (indoor and outdoor) and cow-bait. Morphological characters were used to initially identify the mosquitoes, and then, species were further separated by multiplex PCR. Four species were identified in the An. barbirostris complex: An. wejchoochotei Taai & Harbach (Diptera: Culicidae), An. saeungae Taai & Harbach (Diptera: Culicidae), An. dissidens Taai & Harbach (Diptera: Culicidae), and An. campestris Reid (Diptera: Culicidae). The species showed exophagic and zoophilic behavior, except An. wejchoochotei in Chanthaburi, which displayed slight endophilic behavior. Indoor and outdoor blood feeding behavior of An. wejchoochotei and An. saeungae peaked at 2000 h. An updated distribution map of the An. barbirostris complex in Thailand is presented. With a better understanding of the sibling vector species and their spatial distributions, more specific and effective control strategies for vectors of malaria and filariasis in Thailand can be achieved.

Review of Issues on Residual Malaria Transmission.

Residual malaria transmission is the actual maintained inoculation of Plasmodium, in spite of a well-designed and implemented vector control programs, and is of great concern for malaria elimination. Residual malaria transmission occurs under several possible circumstances, among which the presence of exophilic vector species, such as Anopheles dirus, or indoor- and outdoor-biting vectors, such as Anopheles nili, or specific behavior, such as feeding on humans indoors, then resting or leaving the house the same night (such as Anopheles moucheti) or also changes in behavior induced by insecticides applied inside houses, such as the well-known deterrent effect of permethrin-treated nets or the irritant effect of DDT. The use of insecticides may change the composition of local Anopheles populations, such as A. arabiensis taking up the place of A. gambiae in Senegal, A. aquasalis replacing A. darlingi in Guyana, or A. harrisoni superseding A. minimus in Vietnam. The change in behavior, such as biting activity earlier than usually reported-for example, Anopheles funestus after a large-scale distribution of long-lasting insecticidal nets-or insecticide resistance, in particular the current spread of pyrethroid resistance, could hamper the efficacy of classic pyrethroid-treated long-lasting insecticidal nets and maintained transmission. These issues must be well documented in every situation to elaborate, implement, monitor, and evaluate tailored vector control programs, keeping in mind that they must be conceived as integrated programs with several well and appropriately coordinated approaches, combining entomological but also parasitological, clinical, and social methods and analyses. A successful integrated vector control program must then be designed to reduce transmission and incidence rates of malaria morbidity and overall mortality.

Evaluation of fecal immunoassays for canine Echinococcus infection in China.

Human echinococcosis is present worldwide but it is in China that disease prevalence is the highest. In western China, especially in the Tibetan Plateau, the burden of echinococcosis is the most important. Dogs are a major definitive host of Echinococcus and monitoring the presence of Echinococcus worms in dogs is therefore essential to efficiently control the disease. Detection kits based on three different technologies including sandwich ELISA, (indirect) ELISA, and gold immunodiffusion, are currently marketed and used in China. The objective of this work was to assess the efficacy of these kits, in particular with respect to sensitivity and specificity. Four fecal antigen detection kits for canine infection reflecting the three technologies were obtained from companies and tested in parallel on 220 fecal samples. The results indicate that the performance is lower than expected, in particular in terms of sensitivity. The best results were obtained with the sandwich ELISA technology. The gold immunofiltration yielded the poorest results. In all cases, further development is needed to improve the performance of these kits which are key components for the control of echinococcosis.

New assessment of Anopheles vector species identification using MALDI-TOF MS.

BACKGROUND: Anopheles species identification is essential for an effective malaria vector control programme. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been developed to identify adult Anopheles species, using the legs or the cephalothorax. The protein repertoire from arthropods can vary according to compartment, but there is no general consensus regarding the anatomic part to be used. METHODS: To determine the body part of the Anopheles mosquitoes best suited for the identification of field specimens, a mass spectral library was generated with head, thorax with wings and legs of Anopheles gambiae, Anopheles arabiensis and Anopheles funestus obtained from reference centres. The MSL was evaluated using two independent panels of 52 and 40 An. gambiae field-collected in Mali and Guinea, respectively. Geographic variability was also tested using the panel from Mali and several databases containing added specimens from Mali and Senegal. RESULTS: Using the head and a database without specimens from the same field collection, the proportion of interpretable and correct identifications was significantly higher than using the other body parts at a threshold value of 1.7 (p < 0.0001). The thorax of engorged specimens was negatively impacted by the blood meal after frozen storage. The addition of specimens from Mali into the database significantly improved the results of Mali panel (p < 0.0001), which became comparable between head and legs. With higher identification scores, the using of the head will allow to decrease the number of technical replicates of protein extract per specimen, which represents a significant improvement for routine use of MALDI-TOF MS. CONCLUSIONS: The using of the head of Anopheles may improve the performance of MALDI-TOF MS. Region-specific mass spectrum databases will have to be produced. Further research is needed to improve the standardization in order to share online spectral databases.

Species diversity and insecticide resistance within the Anopheles hyrcanus group in Ubon Ratchathani Province, Thailand.

BACKGROUND: Members of the Anopheles hyrcanus group have been incriminated as important malaria vectors. This study aims to identify the species and explore the insecticide susceptibility profile within the Anopheles hyrcanus group in Ubon Ratchathani Province, northeastern Thailand where increasing numbers of malaria cases were reported in 2014. METHODS: Between 2013 and 2015, five rounds of mosquito collections were conducted using human landing and cattle bait techniques during both the rainy and dry seasons. Anopheles mosquitoes were morphologically identified and their insecticide susceptibility status was investigated. Synergist bioassays were carried out with An. hyrcanus (s.l.) due to their resistance to all insecticides. An ITS2-PCR assay was conducted to identify to species the Hyrcanus group specimens. RESULTS: Out of 10,361 Anopheles females collected, representing 18 taxa in 2 subgenera, 71.8% were morphologically identified as belonging to the Hyrcanus Group (subgenus Anopheles), followed by An. barbirostris group (7.9%), An. nivipes (6.5%), An. philippinensis (5.9%) and the other 14 Anopheles species. Specimens of the Hyrcanus Group were more prevalent during the rainy season and were found to be highly zoophilic. Anopheles hyrcanus (s.l.) was active throughout the night, with an early peak of activity between 18:00 h and 21:00 h. ITS2-PCR assay conducted on 603 DNA samples from specimens within the Hyrcanus Group showed the presence of five sisters species. Anopheles peditaeniatus was the most abundant species (90.5%, n = 546), followed by An. nitidus (4.5%, n = 27), An. nigerrimus (4.3%, n = 26), An. argyropus (0.5%, n = 3), and An. sinensis (0.2%, n = 1). All An. hyrcanus (s.l.) specimens that were found resistant to insecticides (deltamethrin 0.05%, permethrin 0.75% and DDT 4% and synergist tests) belonged to An. peditaeniatus. The degree of resistance in An. peditaeniatus to each of these three insecticides was approximately 50%. Addition of PBO (Piperonyl butoxide), but not DEF (S.S.S-tributyl phosphotritioate), seemed to restore susceptibility, indicating a potential role of oxidases as a detoxifying enzyme resistance mechanism. CONCLUSIONS: A better understanding of mosquito diversity related to host preference, biting activity and insecticide resistance status will facilitate the implementation of locally adapted vector control strategies.

Stegomyia Indices and Risk of Dengue Transmission: A Lack of Correlation.

Dengue is present in 128 countries worldwide and is still expanding. There is currently no treatment or universally approved vaccine available. Therefore, prevention and control of mosquito vectors remain the most efficient ways of managing the risk of dengue outbreaks. The Stegomyia indices have been developed as quantitative indicators of the risk of dengue outbreaks. However, conflictual data are circulating about their reliability. We report in this article the first extensive study on Stegomyia indices, covering 78 locations of differing environmental and socio-economic conditions, climate, and population density across Indonesia, from West Sumatra to Papua. A total of 65,876 mosquito larvae and pupae were collected for the study. A correlation was found between incidence and human population density. No correlation was found between the incidence of dengue and the Stegomyia indices.