Cherng Shii Leong

Nationwide Distribution of Culex Mosquitoes and Associated Habitat Characteristics at Residential Areas in Malaysia

Abstract A standardized larval dipping method was used to determine the infestation rates of Culex and other species of mosquitoes in stagnant water at 20 residential areas. This study also examined the associations between Culex distribution and various habitat characteristics across all states in Malaysia. Identification of 7,848 specimens yielded 6 species dominated by Culex quinquefasciatus (82.74%), followed by Cx. vishui (14.39%), Cx. gelidus (2.70%), Lutzia fuscanus (0.11%), Armigeres subalbatus (0.05%), and Anopheles separatus (0.01%). The Culex larvae occurred in stagnant water with pH ranging from 6.4 to 8.2; conductivity, 139.7 to 6635.2 µs/cm; salinity, 0.07 to 3.64 ppt; total dissolved solids, 0.09 to 4.27g/liter; and dissolved oxygen, 5.11 to 8.11 mg/liter. The mean number of Culex larvae was positively correlated with pH, conductivity, salinity, and total dissolved solids. In contrast, the elevation and dissolved oxygen were found negatively correlated with mean number of Culex larvae. This study documented baseline information on the habitat characteristics of Culex species for the 1st time at different residential areas in Malaysia. The findings of this study will be a timely reminder to local authorities that effective control measures should be monitored regularly in order to reduce the nuisance of these mosquitoes and the risks of disease transmission.

Enzymatic characterization of insecticide resistance mechanisms in field populations of Malaysian Culex quinquefasciatus Say (Diptera: Culicidae)

Background There has been no comprehensive study on biochemical characterization of insecticide resistance mechanisms in field populations of Malaysian Culex quinquefasciatus. To fill this void in the literature, a nationwide investigation was performed to quantify the enzyme activities, thereby attempting to characterize the potential resistance mechanisms in Cx. quinquefasciatus in residential areas in Malaysia. Methodology/Principal Findings Culex quinquefasciatus from 14 residential areas across 13 states and one federal territory were subjected to esterases, mixed function oxidases, glutathione-S-transferase and insensitive acetylcholinesterase assays. Enzyme assays revealed that α-esterases and β-esterases were elevated in 13 populations and 12 populations, respectively. Nine populations demonstrated elevated levels of mixed function oxidases and glutathione-S-transferase. Acetylcholinesterase was insensitive to propoxur in all 14 populations. Activity of α-esterases associated with malathion resistance was found in the present study. In addition, an association between the activity of α-esterases and β-esterases was also demonstrated. Conclusions/Significance The present study has characterized the potential biochemical mechanisms in contributing towards insecticide resistance in Cx. quinquefasciatus field populations in Malaysia. Identification of mechanisms underlying the insecticide resistance will be beneficial in developing effective mosquito control programs in Malaysia.

Collective behavior quantification on human odor effects against female Aedes aegypti mosquitoes—Open source development

Classifying and quantifying mosquito activity includes a plethora of categories, ranging from measuring flight speeds, repellency, feeding rates, and specific behaviors such as home entry, swooping and resting, among others. Entomologists have been progressing more toward using machine vision for efficiency for this endeavor. Digital methods have been used to study the behavior of insects in labs, for instance via three-dimensional tracking with specialized cameras to observe the reaction of mosquitoes towards human odor, heat and CO2, although virtually none was reported for several important fields, such as repellency studies which have a significant need for a proper response quantification. However, tracking mosquitoes individually is a challenge and only limited number of specimens can be studied. Although tracking large numbers of individual insects is hailed as one of the characteristics of an ideal automated image-based tracking system especially in 3D, it also is a costly method, often requiring specialized hardware and limited access to the algorithms used for mapping the specimens. The method proposed contributes towards (a) unlimited open source use, (b) a low-cost setup, (c) complete guide for any entomologist to adapt in terms of hardware and software, (d) simple to use, and (e) a lightweight data output for collective behavior analysis of mosquitoes. The setup is demonstrated by testing a simple response of mosquitoes in the presence of human odor versus control, one session with continuous human presence as a stimuli and the other with periodic presence. A group of female Aedes aegypti (Linnaeus) mosquitoes are released into a white-background chamber with a transparent acrylic panel on one side. The video feed of the mosquitoes are processed using filtered contours in a threshold-adjustable video. The mosquitoes in the chamber are mapped on the raster where the coordinates of each mosquito are recorded with the corresponding timestamp. The average distance of the blobs within the frames against time forms a spectra where behavioral patterns can be observed directly, whether any collective effect is observed. With this method, 3D tracking will not be required and a more straightforward data output can be obtained.

Aedes aegypti(Linnaeus) larvae from dengue outbreak areas in Selangor showing resistance to pyrethroids but susceptible to organophosphates

The resistance status of Selangor Aedes aegypti (Linnaeus) larvae against four major groups of insecticides (i.e., organochlorines, carbamates, organophosphates and pyrethroids) was investigated. Aedes aegypti were susceptible against temephos (organophosphate), although resistance (RR50 = 0.21-2.64) may be developing. The insecticides susceptibility status of Ae. aegypti larvae were found heterogeneous among the different study sites. Results showed that Ae. aegypti larvae from Klang, Sabak Bernam and Sepang were susceptible against all insecticides tested. However, other study sites exhibited low to high resistance against all pyrethroids (RR50 = 1.19-32.16). Overall, the application of synergists ethacrynic acid, S.S.S.- tributylphosphorotrithioate and piperonyl butoxide increased the toxicity of insecticides investigated. However, the application failed to increase the mortality to susceptible level (>97%) for certain populations, therefore there are chances of alteration of target site resistance involved. Biochemical assays revealed that α-esterase, (Gombak, Kuala Langat, Kuala Selangor and Sabak Bernam strains) β-esterase (Klang and Sabak Bernam strains), acetylcholinesterase (Kuala Selangor and Sabak Bernam strains), glutathione-S-transferase (Kuala Selangor and Sabak Bernam strains) and mono-oxygenases (Gombak, Hulu Langat, Hulu Selangor and Kuala Langat strains) were elevated. Spearman rank-order correlation indicated a significant correlation between resistance ratios of: DDT and deltamethrin (r = 0.683, P = 0.042), cyfluthrin and deltamethrin (r = 0.867, P =0.002), cyflyuthrin and lambdacyhalothrin (r = 0.800, P =0.010), cyfluthrin and permethrin (r = 0.770, P =0.015) deltamethrin and permethrin (r = 0.803, P =0.088), propoxur and malathion (r = 0.867, P = 0.002), malathion and temephos (r = 0.800, P = 0.010), etofenprox and MFO enzyme (r = 0.667, P =0.050). The current study provides baseline information for vector control programs conducted by local authorities. The susceptibility status of Ae. aegypti should be monitored sporadically to ensure the effectiveness of current vector control strategy in Selangor.

Effects of low-powered RF sweep between 0.01-20 GHz on female Aedes Aegypti mosquitoes: A collective behaviour analysis

There are many products claiming to be an electronic solution towards repelling mosquitoes. Several reviews were published in debunking these claims. However, there is a lack of a systematic study on effects of electromagnetic (EM) or more specifically, radio frequency (RF) waves against mosquitoes due to the conclusions made in those years. Therefore, we attempt to establish a fundamental study on female Aedes Aegypti (Linnaeus) mosquitoes by quantifying the collective behavior of the mosquitoes against a continuous stream of low-powered RF signals via a broadband horn antenna using image processing methods. By examining the average lateral and vertical positions of the mosquitoes versus frequency and time, the data shows negligible consistency in the reactions of the mosquitoes toward the different frequencies ranging from 10 to 20,000.00 MHz, with a step of 10 MHz. This was done by examining 33 hours of spatiotemporal data, which was divided into three sessions. All three sessions showed totally different convolutions in the positions in arbitrary units based on the raster scan of the image processing output. Several frequencies apparently showed up to 0.2–70% shift in both lateral and vertical components along the spectrum, without repeatability for all three sessions. This study contributes to the following: A pilot study for establishing the collective effects of RF against mosquitoes, open-source use, and finally a low-cost and easily adaptable platform for the study of EM effects against any insects.