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Development of Insecticide Resistance in Culex Quinquefasciatus Mosquito (Diptera: Culicidae) in India 3

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Development of Insecticide Resistance in Culex Quinquefasciatus Mosquito (Diptera: Culicidae) in India 3 J. Pestic. Sci., 35(1), 000–000 (2010) DOI: 10.1584/jpestics.G09-36 Note phatic filariasis with worldwide distribution. This mosquito is Development of insecticide resistance in present in high density throughout the year due to its high degree Culex quinquefasciatus mosquito of adaptability in its preference for breeding sites, host speci- ficity, and high reproductive potential.1) According to the World (Diptera: Culicidae) from different Health Organization (WHO), 1.3 billion people are at risk of locations in India lymphatic filariasis infections and about 120 million people are affected in 83 countries, of which 45.5 million people live in the Indian subcontinent and 40 million in Sub-Saharan Africa.2,3) Devi Shankar SUMAN, Sachin N. TIKAR, Marked efforts have been made to control this disease along with 4) Brahma Dutta PARASHAR* and Shri PRAKASH vector transmission at various levels ; however, the development of resistance in C. quinquefasciatus against organophosphorous Defence Research and Development Establishment, (OP), pyrethroids and microbial insecticides has been reported in Gwalior-474002, India different parts of the world.5–10) Among mosquitoes, particularly (Received August 8, 2009; Accepted October 5, 2009) Anophelines and Aedes, resistance to various insecticides has been recorded from India and neighbouring countries.11–13) In Culex quinquefasciatus is a domestic mosquito known as a pri- Jodhpur, Bikaner, Jamnagar and Bathinda cantonment areas, con- mary vector of lymphatic filariasis. We report the development of trol programmes have been operated; however, the resistance insecticide resistance in four populations of C. quinquefasciatus level to any group of insecticides has not been monitored. The Say, 1823 in Jodhpur (JD), Bikaner (BKN), Jamnagar (JMN) build up of the high population density of C. quinquefasciatus and Bathinda (BTH) in India in comparison to a susceptible has been observed in these containment areas, which may be due laboratory population against larvicides such as temephos, to the development of insecticide resistance in these areas. Thus, fenthion, Bacillus thuringiensis var. israelensis and Neemarin in consideration of this, the development of insecticide resistance (azadirachtin), and adulticides such as cypermethrin, alpha cyper- in larval and adult C. quinquefasciatus in these areas has been methrin and lambda cyhalothrin. The resistance ratio (RR) at evaluated for OP, B. thuringiensis var. israelensis, neem com- pound—Neemarin and pyrethroids from the point of view of LC50 between field and laboratory populations was used for re- sistance expression. JD strain showed 10.8-, 6.94-, 5.29- and changes in the management strategy of using insecticides to 2.82-fold resistance to temephos, fenthion, Neemarin and cyper- which C. quinquefasciatus populations are susceptible. methrin, respectively. BTH strain showed resistance to temephos, Materials and Methods fenthion, lambda cyhalothrin, alpha cypermethrin and cyperme- thrin (9.06-, 2.06-, 3.33-, 4.96- and 3.19-fold, respectively). RR 1. Insect collection and rearing of BKN strain to temephos, fenthion, Neemarin and alpha cyper- Larval stages of C. quinquefasciatus collected from army stations methrin was 5.17-, 4.12-, 4.33- and 3.04-fold, respectively; how- in Jodhpur (JD) and Bikaner (BKN) (Rajasthan), Bathinda (BTH) ever RR was least in the JMN strain against most insecticides ex- (Punjab), and Air Force station Jamnagar (JMN) (Gujrat) were cept alpha cypermethrin and fenthion. This study suggests that B. transported to the Defence Research and Development Establish- thuringiensis var israelensis and Neemarin may be efficient for ment, Gwalior (Fig. 1). These larvae were maintained under stan- larval control as an alternative to fenthion- and temephos-resist- dard laboratory conditions at 27Ϯ1°C and relative humidity ant strains of C. quinquefasciatus while lambda cyhalothrin and 75Ϯ5%. They were kept at a density of 100 larvae in 2 l de-chlo- cypermethrin may be used effectively as an adulticide of this rinated water in enamel bowls and given yeast tablets as larval mosquito. © Pesticide Science Society of Japan food. After pupation, pupae were transferred to adult cages. Fifty pairs of male and female mosquitoes were released into a mos- Keywords: Culex quinquefasciatus, susceptibility status, OP quito cage (750 mmϫ650 mmϫ650 mm). In the cage, adults insecticides, Bacillus thuringiensis var. israelensis, Neem insecti- were provided with 10% sugar solution ad libitum dispensed cide, pyrethroids. through a cotton wick in small Petri dishes. Twice a week, chicken was offered as a source for blood feeding by female mos- quitoes. Glass Petri dishes with 400 ml water were kept for ovipo- Introduction sition. The egg rafts were transferred to a 2 l enamel bowl for Culex quinquefasciatus Say, 1823 is a primary vector of lym- hatching and further development of larvae to the adult stage. * To whom correspondence should be addressed. 2. Insecticides E-mail: [email protected] The following insecticide formulations were used for susceptibil- Published online XXXXX ity studies: for larval bioassay, Abate® (Temephos 50% EC) © Pesticide Science Society of Japan (American Cynamide, NJ), Baytex® (Fenthion 82.5% EC) (Bayer 2 D. S. Suman et al. Journal of Pesticide Science ing serially diluted insecticide concentrations were used for the study. Knockdown was recorded at 10-min intervals of 1-hr expo- sure and treated mosquitoes were transferred to a holding tube for recovery. Sugar solution (10%) was provided as adult diet. Final mortality was recorded after a 24-hour recovery period. 4. Data analysis Data were subjected to probit analysis (Indostat Statistical Soft- ware, India) to determine the 50% lethal concentration (LC50) and 90% lethal concentration (LC90) for OPs, B. thuringiensis var. israelensis, neem compound and pyrethroids.17) If control mortality ranged from 5 to 20%, the mortality of treated groups was corrected according to Abbott’s formula.18) The RR was cal- culated as the ratio between LC50 of the field strain and LC50 of the laboratory strain of C. quinquefasciatus. Results 1. Larval bioassay The LAB strain of C. quinquefasciatus showed more susceptibil- ϭ ϭ ity to temephos (LC50 0.0015 mg/l) than fenthion (LC50 0.0114 Fig. 1. Locations of collections of Culex quinquefasciatus in India. mg/l). The LC50 of JD, BKN, JMN and BTH strains of C. quin- 1, Bathinda; 2, Bikaner; 3, Jodhpur; 4, Jamnagar; 5, Gwalior. quefasciatus against temephos (Table 1) ranged from 0.0048 mg/l (JMN strain) to 0.0162 mg/l (JD strain) and the LC90 of these strains ranged between 0.0325 mg/l and 0.1125 mg/l, respectively. Ltd., Germany), Neemarin® compound (Azadirachtin 1% EC) The considerable resistance of field strains of this mosquito to and Bacillus thuringiensis var. israelensis strain 164, serotype H- temephos was observed, ranging from 3.2- (JMN strain) to 10.8- 14 WP (Biotech International Ltd., India), and for adult bioassay, fold (JD strain) of the LAB strain. The susceptibility status of cypermethrin 10% EC, alpha cypermethrin 10% EC (Insecticide different field strains of mosquito (Table 1) to fenthion showed India Ltd., India) and lambdacyhalothrin 5% EC (Syngenta Crop the lowest values of LC50 and LC90 (0.0238 mg/l and 0.1314 mg/l, Protection Private Ltd., India). respectively) in the BTH strain and highest (0.0792 mg/l and 0.4343 mg/l, respectively) in JD strain. BTH (2.08-fold) and JD 3. Bioassay (6.94-fold) strains exhibited minimum and maximum levels of re- 3.1. Larval bioassay sistance in comparison to the LAB strain (Table 1). Larval susceptibility assay was carried out according to WHO LC50 and LC90 of the LAB strain of C. quinquefasciatus guidelines.14) Late 3rd–early 4th instar of the F1 generation in 5 against B. thuringiensis israelensis serotype-14 were 0.0283 mg/l replicates (20 larvae/replicate) in 250 ml dechlorinated water in a and 0.1209 mg/l, respectively. The LC50 and LC90 activity of B. 400 ml glass beaker have been used for susceptibility evaluation. thuringiensis israelensis serotype-14 against larvae of different A 1% stock solution of various insecticidal formulations was pre- field strains of C. quinquefasciatus ranged from 0.0351 to 0.0565 pared in distilled water and fresh stock solution was used to make mg/l and 0.1108 to 0.1467 mg/l, respectively (Table 1). subsequent 5–6 serial dilutions. All experimental beakers were The susceptibility status of larvae of different strains of this kept under a 12-hr light : dark cycle. Mortality was recorded after species to Neemarin showed a wide range of LC50 (0.0137 to 24-hr exposure to temephos, fenthion, Neemarin and B. 0.0726 mg/l) and LC90 (4.2514 to 13.67 mg/l) after 24-hr expo- thuringiensis var. israelensis insecticides. Moribund larvae were sure, showing an increase in resistance from 1.35-fold (JMN considered to be dead. strain) to 5.29-fold (JD strain) in comparison to the LAB strain (Table 1). 3.2 Adult bioassay The adult bioassay was carried out using WHO test kits accord- 2. Adult bioassay ing to WHO methods under standard laboratory conditions.15) The susceptibility status of adults of LAB, JD, BKN, JMN and Two- to four-day-old non-blood-fed 20 females of F1 generation BTH strains of C. quinquefasciatus to various pyrethroids is in 5 replicates were used for toxicity studies. Acetone was used to shown in Table 2. LC50 and LC90 of adults of these strains against prepare 1% stock solution and serial dilutions. To prepare 1% in- lambda cyhalothrin ranged from 0.0085 to 0.0293% and 0.0450 secticide-treated paper, a 12 cmϫ15 cm Whatman filter paper no. to 0.1708%, respectively, with 0.96- to 3.33-fold higher resist- 1 is treated with 2.75 ml insecticide solution (2.42 mg/ml a.i. ance than the LAB strain of C. quinquefasciatus.
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