Species abundance and density of malaria vectors in Western Thailand and implications for disease transmission.

Understanding the dynamics of malaria vectors and their interactions with environmental factors is crucial for effective malaria control. This study investigated the abundance, species composition, seasonal variations, and malaria infection status of female mosquitoes in malaria transmission and non-transmission areas in Western Thailand. Additionally, the susceptibility of malaria vectors to pyrethroid insecticides was assessed. Entomological field surveys were conducted during the hot, wet, and cold seasons in both malaria transmission areas (TA) and non-transmission areas (NTA). The abundance and species composition of malaria vectors were compared between TA and NTA. The availability of larval habitats and the impact of seasonality on vector abundance were analyzed. Infection with Plasmodium spp. in primary malaria vectors was determined using molecular techniques. Furthermore, the susceptibility of malaria vectors to pyrethroids was evaluated using the World Health Organization (WHO) susceptibility test. A total of 9799 female mosquitoes belonging to 54 species and 11 genera were collected using various trapping methods. The number of malaria vectors was significantly higher in TA compared to NTA (P < 0.001). Anopheles minimus and An. aconitus were the predominant species in TA, comprising over 50% and 30% of the total mosquitoes collected, respectively. Seasonality had a significant effect on the availability of larval habitats in both areas (P < 0.05) but did not impact the abundance of adult vectors (P > 0.05). The primary malaria vectors tested were not infected with Plasmodium spp. The WHO susceptibility test revealed high susceptibility of malaria vectors to pyrethroids, with mortality rates of 99-100% at discriminating concentrations. The higher abundance of malaria vectors in the transmission areas underscores the need for targeted control measures in these regions. The susceptibility of malaria vectors to pyrethroids suggests the continued effectiveness of this class of insecticides for vector control interventions. Other factors influencing malaria transmission risk in the study areas are discussed. These findings contribute to our understanding of malaria vectors and can inform evidence-based strategies for malaria control and elimination efforts in Western Thailand.

Demonstration of RNAi Yeast Insecticide Activity in Semi-Field Larvicide and Attractive Targeted Sugar Bait Trials Conducted on Aedes and Culex Mosquitoes.

Eco-friendly new mosquito control innovations are critical for the ongoing success of global mosquito control programs. In this study, Sh.463_56.10R, a robust RNA interference (RNAi) yeast insecticide strain that is suitable for scaled fermentation, was evaluated under semi-field conditions. Inactivated and dried Sh.463_56.10R yeast induced significant mortality of field strain Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus larvae in semi-field larvicide trials conducted outdoors in St. Augustine, Trinidad, where 100% of the larvae were dead within 24 h. The yeast was also stably suspended in commercial bait and deployed as an active ingredient in miniature attractive targeted sugar bait (ATSB) station sachets. The yeast ATSB induced high levels of Aedes and Culex mosquito morbidity in semi-field trials conducted in Trinidad, West Indies, as well as in Bangkok, Thailand, in which the consumption of the yeast resulted in adult female mosquito death within 48 h, faster than what was observed in laboratory trials. These findings support the pursuit of large-scale field trials to further evaluate the Sh.463_56.10R insecticide, a member of a promising new class of species-specific RNAi insecticides that could help combat insecticide resistance and support effective mosquito control programs worldwide.

The impact of insect growth regulators on adult emergence inhibition and the fitness of Aedes aegypti field populations in Thailand.

BACKGROUND: The frequent use of insecticides in vector control causes the development of insecticide resistance. Insect growth regulators (IGRs), which effect insect development, are used as a promising alternative to control resistant insect vectors. This study aimed to develop novel effective tools for Aedes aegypti control by evaluating the efficacy of different IGRs on larval development, blood feeding capacity, fecundity, and fertility in females and sperm productivity in males across geographical regions of Thailand. METHODS: The efficacy of 16 technical grade IGRs were evaluated against laboratory strain Ae. aegypti larvae in order to determine their emergence inhibition (EI) at 50% and 95% under laboratory conditions. Six IGRs were selected for fecundity, fertility, and sperm productivity studies using feed-through treatments at EI95 concentration levels against adult Ae. aegypti field strains. RESULTS: The results from larval bioassay tests indicate that juvenile hormone mimics (EI50 = 0.010-0.229 ppb; EI95 = 0.066-1.118 ppb) and chitin synthesis inhibitors affecting CHS1 (EI50 = 0.240-2.412 ppb; EI95 = 0.444-4.040 ppb) groups effectively inhibited adult Ae. aegypti emergence. Methoprene and fenoxycarb significantly reduced blood feeding capacity. Egg production was comparable among strains while methoprene, pyriproxyfen and diflubenzuron induced egg production. Egg retention was detected in females fed on diflubenzuron. Methoprene, fenoxycarb, diflubenzuron, and teflubenzuron reduced egg hatching rates in mosquito field strains compared to laboratory strain. Male mosquitoes fed on fenoxycarb showed significantly lower sperm production compared to other treatments. CONCLUSION: Juvenile hormone analogues and chitin synthesis inhibitors affecting CHS1 groups showed excellent results in adult emergence inhibition in this study. They also disrupted reproductive systems in both adult males and females. This study suggested that they can be used as an alternative larvicide in mosquito control programs.

Optimization of Culture Protocols to Isolate Leptospira spp. from Environmental Water, Field Investigation, and Identification of Factors Associated with the Presence of Leptospira spp. in the Environment.

The successful culture of Leptospira spp. from the environment is challenging. Here, we optimized the isolation of Leptospira spp. from water samples spiked with different species and initial concentrations of this organism. The time periods between water sampling and the isolation process were varied (0, 2, and 4 weeks). Bacterial cultures were observed under a microscope, and cultures were graded for cell density, weekly, for 12 weeks. Most pathogenic Leptospira spp. were difficult to culture under all conditions. All conditions of water samples spiked with novel species of Leptospira subclade P1 were culture positive within 2 weeks. For Leptospira subclade P2, storing samples for 2 weeks prior to isolation resulted in more successful isolation compared with isolation after other storage conditions. For subclade S1, all samples with initial bacterial concentrations of more than 103 colonies/mL, under all storage conditions, were successfully cultured. These results suggest that storing contaminated water samples for 2 to 4 weeks in the dark at an ambient temperature prior to culturing can improve the isolation of Leptospira spp. from the samples. We implemented this protocol and collected water samples from natural sources accessed by both humans and animals. Leptospira spp. was identified in 32% (35/109) of water samples. The animal species using a water source influenced the likelihood of water samples being contaminated with Leptospira spp. Cultures of Leptospira spp. from environmental samples can provide useful information for understanding the complex interactions between humans, animals and the environment in the transmission of leptospirosis.

Entomological Surveillance for Zika and Dengue Virus in Aedes Mosquitoes: Implications for Vector Control in Thailand.

Entomological surveillance for arthropod-borne viruses is vital for monitoring vector-borne diseases and informing vector control programs. In this study, we conducted entomological surveillance in Zika virus endemic areas. In Thailand, it is standard protocol to perform mosquito control within 24 h of a reported dengue case. Aedes females were collected within 72 h of case reports from villages with recent Zika-human cases in Kamphaeng Phet Province, Thailand in 2017 and 2018. Mosquitoes were bisected into head-thorax and abdomen and then screened for Zika (ZIKV) and dengue (DENV) viruses using real-time RT-PCR. ZIKV RNA was detected in three samples from two female Ae. aegypti (1.4%). A partial envelope sequence analysis revealed that the ZIKV sequences were the Asian lineage identical to sequences from ZIKV-infected cases reported in Thailand during 2016 and 2017. Dengue virus-1 (DENV-1) and dengue virus-4 (DENV-4) were found in four Ae. aegypti females (2.8%), and partial capsid sequences were nearly identical with DENV-1 and DENV-4 from Thai human cases reported in 2017. Findings in the current study demonstrate the importance of entomological surveillance programs to public health mosquito-borne disease prevention measures and control.