Irritant and repellent behaviors of sterile male Aedes aegypti (L.) (Diptera: Culicidae) mosquitoes are crucial in the development of disease control strategies applying sterile insect technique.

The mosquito Aedes aegypti, known to transmit important arboviral diseases, including dengue, chikungunya, Zika and yellow fever. Given the importance of this disease vector, a number of control programs have been proposed involving the use of the sterile insect technique (SIT). However, the success of this technique hinges on having a good understanding of the biology and behavior of the male mosquito. Behavioral responses of Ae. aegypti male populations developed for SIT technology were tested under laboratory conditions against chemical and natural irritants and repellents using an excito-repellency (ER) chamber. The results showed that there were no significant behavioral escape responses in any of the radiation-sterilized male Ae. aegypti test populations when exposed to citronella, DEET, transfluthrin, and deltamethrin, suggesting that SIT did not suppress the expected irritancy and repellency (avoidance) behaviors. The type of information reported in the current study is vital in defining the effects of SIT on vector behavior and understanding how such behavior may influence the success of SIT technology with regard to other vector control interventions.

Life-history traits of Aedes aegypti (Linnaeus) distributed across a latitudinal range of 23 °N-6 °S.

The growth and associated traits of Aedes aegypti (L.) mosquitoes may adapt and evolve in response to the costs associated with body size in relation to latitudinal variation. We analyzed the life-history traits and energy reserves of field-collected mosquitoes from Taiwan, Thailand, and Indonesia along a latitudinal range spanning from 23°N to 6°S. A U-shaped relationship between body size and latitude was observed. Our study demonstrated the role of latitudinal temperature variations in determining the body size patterns of Ae. aegypti. Notably, the body size of the northern populations (from Taiwan) was significantly larger than those of the tropical populations from Thailand and Indonesia. Models have demonstrated that regional precipitation levels may contribute to body trait variations in certain high-latitude populations in Thailand. However, Indonesian populations have high development rates and large body sizes, indicating the involvement of other physiological traits in determining mosquito body size. The reproductive output of the adult females in this study was positively correlated with body size, but our measure of longevity did not covary significantly with the body size. By contrast, the reproductive output of mosquito-tested populations was in inverse proportion to longevity. Additionally, the mean teneral glycogen levels in the Indonesian and Thai populations were 2.5 times higher than those in the Taiwanese populations. The Indonesian and Thai populations had low mean generation and doubling times, resulting in a high intrinsic rate of increase compared with that of the Taiwanese populations, despite the Taiwanese populations having the highest net reproduction rate.

Synergistic Behavioral Response Effect of Mixtures of Andrographis paniculata, Cananga odorata, and Vetiveria zizanioides against Aedes aegypti (Diptera: Culicidae).

Each binary mixture formulation of Vetiveria zizanioides (L.) Nash (VZ) with Andrographis paniculata (Burm.f.) Wall. ex Nees (AP) or Cananga odorata (Lam.) Hook.f. & Thomson (CO) and AP with CO at 1:1, 1:2, 1:3, and 1:4 ratios (v:v) was investigated for behavioral responses on laboratory and field strains of Aedes aegypti. Irritant and repellent activities of each formulation were compared with N,N-diethyl-3-methylbenzamide (DEET) using an excito-repellency test system. The result demonstrated that the mixture of VZ:AP in all combination ratios was the most effective in inducing an irritancy response against the laboratory strain (56.57-73.33%). The highest percentage of escaped mosquitoes exposed to the mixture at a 1:4 ratio (73.33%) was significantly different from DEET (26.67%) (p < 0.05). Against the field strain, the strongest escape response of AP:CO at a 1:1 ratio in the contact trial (70.18%) was significantly different compared with DEET (38.33%) (p < 0.05). There was a weak non-contact escape pattern in all combinations of VZ:CO against the laboratory strains (6.67-31.67%). These findings could lead to the further development of VZ and AP as active ingredients in a repellent that could advance to human use trials.

Comparing Light-Emitting-Diodes Light Traps for Catching Anopheles Mosquitoes in a Forest Setting, Western Thailand.

Light traps are a common method for attracting and collecting arthropods, including disease vectors such as mosquitoes. Various types of traps have been used to monitor mosquitoes in a forest in Western Thailand. In this study, four Light Emitting Diodes (LED) light sources (UV, blue, green, and red) and two fluorescent lights (white and UV) were used to trap nocturnal adult mosquitoes. These traps were used with light alone and not any additional attractant. The experiment was conducted from 18:00 to 06:00 h. on six consecutive nights, every two months, across dry, wet, and cold seasons. All specimens were first identified by morphological features and subsequently confirmed by using PCR. We collected a total of 873 specimens of 31 species in four genera, Anopheles, Aedes, Culex, and Armigeres. Anopheles harrisoni was the predominant species, followed by Aedes albopictus, Culex brevipalpis, Culex nitropunctatus, and Armigeres (Leicesteria) longipalpis. UV fluorescent light was the most effective light source for capturing forest mosquitoes, followed by UV LED, blue LED, green LED, white fluorescent, and red LED. The optimal times for collection were from 21:00 to 03:00 h in the dry season. Our results demonstrate that appropriate sampling times and light sources should be selected for optimal efficiency in vector surveillance programs.

Avoidance Behavior to Guava Leaf Volatile Oil by Three Medically Important Mosquito Vectors.

Volatile organic compounds from various plants have received popular interest as one of the vector control tools due to their eco-friendliness and insect-repellent activities. In this study, an excito-repellency assay system was used to examine the noncontact repellency, contact excitation, and knockdown (KD) effects of guava leaf (Psidium guajava L.) oil against Anopheles minimus (Theobald), Anopheles epiroticus (Linton & Harbach), and Culex quinquefasciatus (Say). The organic components of guava oil were identified by gas chromatography-mass spectrometry analysis with dl-limonene (17.4%), cymene (5.49%), and α-terpinene (5.20%) as the major constituents. At concentrations of 2.5 and 5.0%, 100% escape of An. minimus was recorded in the contact assay and 96-98% escape in the noncontact assay. Guava oil stimulated potent irritant (92% escape) and repellent (61-86% escape) effects against Cx. quinquefasciatus. A lower repellency action was observed against An. epiroticus (17-20% escape). No KD effect was observed for guava oil against An. minimus and Cx. quinquefasciatus at any concentration. However, An. epiroticus was more prone to KD effects, with the highest percentage KD (44% in nonescape group) observed with 5.0% guava oil in the noncontact assay. Mortalities of 35% and 11% were observed for An. epiroticus in the nonescape groups in the contact and noncontact assays, respectively. Concentrations of 1.0% and 2.5% guava oil led to <2% mortality in An. minimus. Our findings highlight guava oil as a promising plant-based mosquito repellent that can be included in insecticide formulations for future mosquito control programs.

Effects of piperonyl butoxide synergism and cuticular thickening on the contact irritancy response of field Aedes aegypti (Diptera: Culicidae) to deltamethrin.

BACKGROUND: Exploiting indoor-resting mosquitoes' innate behavioral responses to commonly used insecticide is crucial in vector control programs. Indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) treated with pyrethroids have become widely used for controlling dengue fever vectors. The present study tested the effects of piperonyl butoxide (PBO) synergism and cuticular thickening on the contact irritancy response of field A. aegypti (Diptera: Culicidae) to deltamethrin in Taiwan and Thailand. RESULTS: The escape response of field mosquitoes treated with PBO was significantly elicited, with an escape percentage increase between 2- and 10-fold. In addition, the escape time was significantly lower in PBO-pretreated mosquitoes compared with field mosquitoes treated with deltamethrin alone. PBO-pretreated mosquitoes from seven out of 11 field strains exhibited a knockdown percentage of 11.23-54.91%, significantly higher than that of mosquitoes in corresponding strains treated with deltamethrin only. The Annan, Zhongxi, Sanmin, and North strains exhibited weak knockdown responses (≤3.75%). The mortality of PBO-pretreated field mosquitoes increased 2- to 75-fold compared with those treated with deltamethrin alone (mortality: 0-6.70%). Furthermore, the effect of cuticular thickness on the escape response of field mosquitoes was significant, that is, the escape response marginally increased inversely to cuticular thickness. By contrast, cuticular thickness was not significantly associated with knockdown or mortality percentage. CONCLUSION: Irritant behavior in mosquitoes was significantly elicited by PBO synergism. PBO incorporating deltamethrin IRS or LLINs may be effective for controlling dengue fever vectors. © 2021 Society of Chemical Industry.

Forced Egg Laying Method to Establish F1 Progeny from Field Populations and Laboratory Strains of Anopheles Mosquitoes (Diptera: Culicidae) in Thailand.

Successful monitoring of physiological resistance of malaria vectors requires about 150 female mosquitoes for a single set of tests. In some situations, the sampling effort is insufficient due to the low number of field-caught mosquitoes. To address this challenge, we demonstrate the feasibility of using the forced oviposition method for producing F1 from field-caught Anopheles mosquitoes. A total of 430 and 598 gravid Anopheles females from four laboratory strains and five field populations, respectively, were tested. After blood feeding, gravid mosquitoes were individually introduced into transparent plastic vials, containing moistened cotton balls topped with a 4 cm2 piece of filter paper. The number of eggs, hatching larvae, pupation, and adult emergence were recorded daily. The mean number of eggs per female mosquito ranged from 39.3 for Anopheles cracens to 93.6 for Anopheles dirus in the laboratory strains, and from 36.3 for Anopheles harrisoni to 147.6 for Anopheles barbirostris s.l. in the field populations. A relatively high egg hatching rate was found in An. dirus (95.85%), Anopheles minimus (78.22%), and An. cracens (75.59%). Similarly, a relatively high pupation rate was found for almost all test species ranging from 66% for An. minimus to 98.7% for Anopheles maculatus, and lowest for An. harrisoni (43.9%). Highly successful adult emergence rate was observed among 85-100% of pupae that emerged in all tested mosquito populations. The in-tube forced oviposition method is a promising method for the production of sufficient F1 progeny for molecular identification, vector competence, insecticide resistance, and bioassay studies.

The Efficacy of Ultrasonic Pest Repellent Devices against the Australian Paralysis Tick, Ixodes holocyclus (Acari: Ixodidae).

Ultrasonic pest repellers are often promoted as a means of protecting people and pets from the bites of hematophagous arthropods, such as ticks. However, to date, there has been no published research on the effectiveness of these devices against the Australian paralysis tick, Ixodes holocyclus Neumann. The purpose of this study was to test the effectiveness of nine ultrasonic devices against female I. holocyclus. Two arenas were constructed, one for the test (with the ultrasonic device) and one for the control (no device). Each arena had a test and an escape chamber, connected by a corridor. Twenty ticks were placed in each test chamber. After the ultrasonic device was operated for 1 h, the number of ticks in both chambers was recorded. Ten replicates were conducted for each device. The average number of ticks that moved from the test to the escape chamber was greater in all the test arenas, with three devices being statistically different from the control. However, the highest percent of ticks that escaped was only 19.5%. This amount is insufficient to offer adequate protection against tick bites and this study adds further weight to previous investigations that ultrasonic devices should not be employed in pest management.

Repellency and Contact Irritancy Responses of Aedes aegypti (Diptera: Culicidae) Against Deltamethrin and Permethrin: A Cross-Regional Comparison.

Control strategies exploiting the innate response of mosquitoes to chemicals are urgently required to complement existing traditional approaches. We therefore examined the behavioral responses of 16 field strains of Aedes aegypti (L.) from two countries, to deltamethrin and permethrin by using an excito-repellency (ER) test system. The result demonstrated that the escape percentage of Ae. aegypti exposed to pyrethroids did not vary significantly between the two countries in both contact and noncontact treatment despite the differing epidemiological patterns. Deltamethrin (contact: 3.57 ± 2.06% to 31.20 ± 10.71%; noncontact: 1.67 ± 1.67% to 17.31 ± 14.85%) elicited relatively lower responses to field mosquitoes when compared with permethrin (contact: 16.15 ± 4.07% to 74.19 ± 4.69%; noncontact: 3.45 ± 2.00% to 41.59 ± 6.98%) in contact and noncontact treatments. Compared with field strains, the mean percentage of escaping laboratory susceptible strain individuals were significantly high after treatments (deltamethrin contact: 72.26 ± 6.95%, noncontact: 61.10 ± 12.31%; permethrin contact: 78.67 ± 9.67%, noncontact: 67.07 ± 7.02%) and the escaped individuals spent significantly shorter time escaping from the contact and noncontact chamber. The results indicated a significant effect of resistance ratio on mean escape percentage, but some strains varied idiosyncratically compared to the increase in insecticide resistance. The results also illustrated that the resistance ratio had a significant effect on the mortality in treatments. However, the mortality in field mosquitoes that prematurely escaped from the treated contact chamber or in mosquitoes that stayed up to the 30-min experimental period showed no significant difference.

Detection of Anaplasma spp. and Bartonella spp. from wild-caught rodents and their ectoparasites in Nakhon Ratchasima Province, Thailand.

The objective of this study was to investigate evidence of emerging anaplasmosis and bartonellosis in rodents from endemic areas of Nakhon Ratchasima, Thailand. Rodent trapping was undertaken in 13 sub-districts of Muang District. The live-capture traps were set up in three locations of selected scrub typhus patient houses for three consecutive nights. Wild-caught rodent whole blood samples and associated ticks and fleas were collected and tested for Anaplasma spp. and Bartonella spp. In addition, heat maps using GIS software were used to determine the density of infection of positive wild-caught rodents. A total of 347 wild-caught rodents of nine species was captured. Rattus rattus (38.6%) was the dominant species. A total of 1,518 Heamaphysalis bandicota ticks and 57 Xenopsylla cheopis fleas was removed. Twenty-two of the 347 tested blood samples (6.3%) were Anaplasma bovis-positive and 121 blood samples and five out of 27 pools of X. cheopis fleas were Bartonella queenslandensis-positive. Of these infected rodents, dual-infections between A. bovis and B. queenslandensis were found in three B. indica rodents. Our results offer new information concerning the infections of A. bovis and B. queenslandensis in both rodents and their ectoparasites collected in high-risk areas of rodent-borne diseases in Thailand.

Excito-repellency Activity of Andrographis paniculata (Lamiales: Acanthaceae) Against Colonized Mosquitoes.

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.

SPECIES DIVERSITY AND SEASONALITY OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) IN SATUN PROVINCE, THAILAND.

Leishmaniasis is prevalent mainly in the southern provinces of Thailand where sand flies are considered to be an important vector. Sand flies were collected using Centers for Disease Control (CDC) light traps in Satun Province from June 2013 to July 2014. A total of 1,982 sand flies (1,228 females and 754 males) were collected. Only female sand flies were identified to the species level and were tested for Leishmania infection using polymerase chain reaction (PCR). Morphological identification revealed 2 genera and 9 species: Phlebotomus stantoni, P. argentipes, Sergentomyia gemmea, S. indica, S. barraudi, S. iyengari, S. bailyi, S. perturbans, and S. silvatica. S. gemmea (57.2%) was the most abundant species. The diversity of sand flies was highest in Thung Wa District. The sand flies were most abundant late in the hot season and early in the rainy season (April to June). The highest number of sand flies was collected in June. Significant correlations between the number of female sand flies and rainfall and between S. gemmea and rainfall were found. Of the female sand flies tested, none were positive for Leishmania spp.