The Barbirostris Complex comprises six formally described species that cannot be differentiated based on morphology alone. Out of these six species, two have been reported as putative malaria vectors, An. campestris and An. wejchoochotei. Five species are present in Thailand, An. barbirostris, An. campestris, An. dissidens, An. saeungae and An. wejchoochotei, while An. vanderwulpi occurs in Indonesia. As these species cannot be accurately differentiated by morphological characters, there is a crucial lack of information on their bionomics and role in the transmission of malaria and filariasis agents. For differentiating the six species, an allele-specific amplification (AS-PCR) based on the second internal transcribed spacer (ITS2) sequence was developed. From 862 mosquitoes in the Barbirostris Complex collected in 23 provinces throughout Thailand, the AS-PCR was able to identify five species and its validation was undertaken on 185 specimens. This multiplex-PCR assay is potentially able to definitely identify all six species of the Barbirostris Complex and was validated on five species present in Thailand.

Airborne spatial repellency (SR) is characterized and distinguished from other chemical actions including contact locomotor excitation and toxicity. The use of volatile spatial repellents is a potential new intervention class for combatting mosquito-borne pathogen transmission; therefore, continuing investigations on the actions of these chemicals that modify mosquito host‐seeking behavior (i.e., bite prevention) is needed. The objective of this study is to characterize the key behavioral avoidance actions of transfluthrin (TFT) to advance spatial repellent development into practical products. Behavioral avoidance responses were observed for adult laboratory strains of Aedes aegypti, Anopheles minimus and An. dirus, and two field populations of An. harrisoni and Ae. aegypti, respectively. Established TFT sublethal (LC₅₀ and LC₇₅), lethal concentrations (LC₉₉) and discriminating concentrations (DCs) were selected corresponding to each mosquito test species. Spatial repellency and contact excitation (‘irritancy’) responses on adult mosquitoes to TFT were assessed using an excito-repellency assay system. At LC₅₀, TFT exhibited strong avoidance with An. minimus (60.1% escape) and An. dirus (80% escape) laboratory strains, showing between 12 and 16x greater escape response than Ae. aegypti (5% escape). Repellency responses for field collected Ae. aegypti and An. harrisoni were 54.9 and 47.1% escape, respectively. After adjusting the initial contact escape response (a measure of combined irritancy and repellency) to estimate only escape due to contact, the LC₅₀ and LC₉₉ showed moderate escape irritancy with laboratory Ae. aegypti (41.4% escape) and no contact activity against the field population. Adjustment showed only weak contact activity (16.1% escape) in laboratory An. minimus at LC₅₀. Spatial repellency is the predominant mode of action of TFT among colonized and field mosquitoes used in this study. Established baseline (susceptible) dose-response curves assist in optimizing SR products for mosquito control and pathogen transmission prevention.

Background: The entomological inoculation rate (EIR) is one of the key indices used to evaluate malaria transmission and vector control interventions. One of the components of the EIR is the sporozoite rate in Anopheles vectors. A systematic review and meta-analysis was performed to identify the prevalence of Plasmodium spp. in field-collected Anopheles species across Thailand. Methods: This systematic review was registered under the PROSPERO number CRD42021297255. Studies that focused on the identification of Plasmodium spp. in Anopheles mosquitoes were identified from the electronic databases PubMed, Web of Science, and Scopus. The quality of the identified studies was determined using the Strengthening the Reporting of Observational Studies in Epidemiology approach. The proportion of Anopheles mosquitoes collected, Anopheles vectors for Plasmodium species, and specificity of Anopheles vectors for Plasmodium species were analyzed. The pooled prevalence of Plasmodium species among the primary vectors (Anopheles dirus, Anopheles minimus, and Anopheles maculatus) was estimated using the random-effects model. Results: Of the 1113 studies identified, 31 were included in the syntheses. Of the 100,910 Anopheles mosquitoes identified for species and sibling species, An. minimus (40.16%), An. maculatus (16.59%), and Anopheles epiroticus (9.18%) were the most prevalent Anopheles species. Of the 123,286 Anopheles mosquitoes identified, 566 (0.46%) were positive for Plasmodium species. The highest proportions of Plasmodium species were identified in Anopheles hodgkini (2/6, 33.3%), Anopheles nigerrimus (2/24, 8.33%), Anopheles balabacensis (4/84, 4.76%), An. dirus (114/4956, 2.3%), Anopheles annularis (16/852, 1.88%), Anopheles kochi (8/519, 1.54%), Anopheles vagus (3/215, 1.4%), and Anopheles baimaii (1/86, 1.16%). The pooled prevalence of Plasmodium species identified in the main Anopheles vectors was 0.4% of that of Plasmodium species identified in An. dirus was 2.1%, that of Plasmodium species identified in An. minimus was 0.4%, and that of Plasmodium species identified in An. maculatus was 0.4%. Conclusions: We found a low prevalence of Plasmodium infection in Anopheles mosquitoes across Thailand. Therefore, the use of EIR to determine the impact of vector control intervention on malaria parasite transmission and elimination in Thailand must be undertaken with caution, as a large number of Anopheles specimens may be required. Graphical

The development of resistance by mosquitoes to current synthetic compounds has resulted in reduced effectiveness of prevention and control methods worldwide. An alternative nonchemical based control tools are needed to be evaluated particularly plant-derived essential oils. Several components of vetiver oil have been documented as insect repellents. However, detailed knowledge of those components action against insect remains unknown. In this study, behavioral response of Anopheles minimus to four constituents of vetiver oil (valencene, terpinen-4-ol, isolongifolene, vetiverol) was evaluated by using the high-throughput screening assay system. Vetiverol and isolongifolene exhibited strong contact irritancy action at 1.0% (80.2% escaping) and 5.0% (81.7% escaping) concentration, respectively, while moderate action was found in both valencene and terpinen-4-ol at 5.0% (57.6% escaping). Only at 1.0% (0.7 spatial activity index [SAI]) and 5.0% (1.0 SAI) of valencene and 0.5% (0.7 SAI) of isolongifolene showed spatial repellency activity. High mortality (58.9–98.2%) was recorded in all concentration of vetiverol and isolongifolene. Meanwhile, valencene exhibited high mortality only at 5.0%, terpinen-4-ol showed very low toxic action (0–4.3%) in all concentration. These proved that valencene in vetiver oil is the promising constituent that can be developed as an alternative mosquito control mean in efforts to prevent disease transmission.

Excito-repellency activity of plant extracts have been increasingly studied as mosquito repellents. In this study, the crude extract of Andrographis paniculata was evaluated for its noncontact repellency, contact excitation (irritancy + repellency), and knockdown/toxicity response against five colonized mosquitoes; Aedes aegypti (L.), Aedes albopictus (Skuse), Anopheles dirus Peyton & Harrison, Anopheles epiroticus Linton & Harbach, and Culex quinquefasciatus Say (Diptera: Culicidae) using an excito-repellency assay system under laboratory-controlled conditions. The escape responses were observed at four different concentrations (0.5–5.0% w/v) with A. paniculata showing strong spatial repellency against Ae. albopictus (96.7% escape) and Ae. aegypti (71.7% escape) at the 2.5% and 0.5% concentrations, respectively. At 0.5% and 5.0% concentrations, the greatest repellency was seen for An. dirus (48.2% escape) and Cx. quinquefasciatus (59.7% escape), respectively. Comparatively, low repellency action was observed against An. epiroticus (1.6–15.0% escape). Escape in contact assays (before adjustment) was generally less pronounced compared to noncontact spatial repellency, with Ae. albopictus showing highest percent escape (71.4% escape) in the contact assay at 1.0% concentration. After adjusting for spatial repellency, escape due to contact irritancy alone was either not present or an insignificant contribution to the overall avoidance response for all species. No knockdown or mortality at 24-h postexposure was observed in any trials. These findings indicate that the A. paniculata crude extract is more active against day-biting mosquitoes; however, this may be a reflection of the time of testing. This study demonstrates compelling evidence that A. paniculata extract performs primarily as a spatial repellent. Further investigations exploring the use A. paniculata as a potential active ingredient in repellent products are needed.