Dhiraj Saha

Insecticide resistance mechanisms in three sucking insect pests of tea with reference to North-East India: an appraisal

Tea is prepared from the tender leaves and buds of Camellia sinensis (L.). In sub-Himalayan tea plantations of North-East India, different management practices are followed to protect the tea crop against different sucking insect pests such as Helopeltis theivora (Hemiptera: Miridae), Empoasca flavescens (Homoptera: Cicadellidae) and Scirtothrips dorsalis (Thysanoptera: Thripidae). Most plantations are managed conventionally through the use of different organo-synthetic insecticides, whereas some are managed organically by using herbal and microbial insecticides. In conventional tea plantations, organo-synthetic insecticides of different functional groups (organochlorines, organopho- sphates, synthetic pyrethroids and neonicotinoids) are routinely applied round the year to keep the sucking insect pest populations under control. A variety of defence mechanisms, including enzymatic detoxification systems (carboxylesterases, glutathione S-transferases and cytochrome P450 monooxygenases), physiological tolerance and behavioural avoidance, protect insect herbivores from these hazardous compounds. Insect pests have evolved mechanisms to degrade metabolically (enzymatically) or otherwise circumvent the toxic effect of many types of chemicals synthesized as modern insecticides. The extent to which insects can metabolize and thereby degrade these toxic or otherwise detrimental chemicals is of considerable importance for their survival in an unfriendly chemical environment. These mechanisms continue to evolve as insects attempt to colonize new plant species or encounter newer molecules of synthetic insecticides. The level and type of detoxifying mechanisms differs greatly, which, therefore, results in varying toxicity among different stages, species and populations. Variation in detoxifying enzyme activity is responsible, in part, at least for the selective toxicity of different insecticides, the development of resistance to insecticides and the selection of host plants. Overexpression of these detoxifying enzymes, capable of metabolizing insecticides, can result in higher levels of metabolic tolerance/resistance to synthetic insecticides.

Insecticide susceptibility and activity of major detoxifying enzymes in female Helopeltis theivora (Heteroptera: Miridae) from sub-Himalayan tea plantations of North Bengal, India

Despite the continuous use of synthetic insecticides during the last two decades, the tea mosquito bug Helopeltis theivora Waterhouse still exists as the most destructive pest of tea in North East India. The susceptibility levels of the female sucking bug collected from conventional (synthetic insecticide treated: Terai and Dooars plain regions) and organic (synthetic insecticide untreated: low-altitude Darjeeling region) tea plantations of the northern part of West Bengal to two synthetic insecticides, quinalphos and cypermethrin, and the activity of three principal detoxifying enzymes were assayed. Compared with the susceptible Darjeeling population, the Terai and Dooars populations showed a resistance factor at the lethal concentrations for 50% level ranging from 547- to 2680.87-fold and from 3810- to 7480-fold for quinalphos and cypermethrin, respectively. General esterases (GEs), glutathione S-transferases (GSTs) and cytochrome P450-mediated mono-oxygenases (CYPs) also showed an increased activity in the Terai and Dooars populations compared with those fromDarjeeling. Defence enzyme activity was enhanced by 15.4- and 17.6-fold for GEs, 1.8- and 1.9-fold for GSTs and 2.1- and 2.4-fold for CYPs in the synthetic insecticide-treated H. theivora populations when compared with the untreated Darjeeling populations. Electrophoretic analysis for GEs showed a higher level of expression for esterase I–VI isozymes in the Terai and Dooars populations when compared with that in the Darjeeling populations. This study reveals a reduced efficacy of quinalphos and cypermethrin against field populations of H. theivora, possibly due to enhanced activities of GEs, GSTs and CYPs. The findings may be used in developing integrated resistance management strategies that can help in the effective control of this major tea pest.

Variation in the Activity of Three Principal Detoxifying Enzymes in Major Sucking Pest of Tea, Helopeltis theivora Waterhouse (Heteroptera: Miridae) from Sub-Himalayan Tea Plantations of West Bengal, India

Helopeltis theivora Waterhouse (Heteroptera: Miridae), is a major sucking pest of tea in North East India along with other tea growing countries. In West Bengal, tea is cultivated in three sub-Himalayan regions, Terai (foothill plains western to river Teesta), the Dooars (foothill plains eastern to river Teesta) and the Darjeeling hill slopes. Most plantations, in these regions are managed conventionally i.e. by spraying different synthetic insecticides and a few by organic farming using different herbal and microbial insecticides. In conventional plantations, continuous application of insecticides may lead to the selection of more tolerant H. theivora populations making the pest difficult to control. So, there is a pressing need to know the biochemical variability in relation to the metabolic resistance in the pest populations and develop a population specific control strategy. Activity of three principal insecticide detoxifying enzymes in H. theivora populations were studied from three tea growing regions of North Bengal. Higher levels of activity of all the studied enzymes were found from conventional tea plantations. In male H. theivora, the activity of general esterases (GEs) was 6.6–11.2 and 10.5–11.4 fold higher, cytochrome P450 (CYPs) was 2.0–3.2 and 3.0–3.2 fold higher and glutathione S-transferase (GSTs) was 5.2–8.3 and 6.4–8.7 fold higher in Terai and the Dooars populations, respectively than organic populations from Darjeeling hill slopes. Similarly, in female H. theivora, activity of GEs was 6.2–10.3 and 8.3–9.6 folds higher, CYPs was 1.9–3.2 and 3.0–3.3 fold, and GSTs was 3.5–5.4 and 4.4–6.0 fold higher in Terai and the Dooars, respectively than the organic populations from Darjeeling hill slopes. The activities of all three enzymes were found to be significantly low in organic plantations from Darjeeling hill slopes. Esterase I–VI isozymes with higher level of expression were found in specimens from conventional tea plantations than organic populations of H. theivora from Darjeeling hill slopes in isozymes study.

Variation in Esterase Activity Among Different Aedes aegypti L. Populations from the Dooars and Terai Regions of West Bengal, India

Mosquitoes are responsible for transmitting the most important vector-borne diseases like malaria, dengue etc. across the world, especially in tropical countries. The most prominent species of the genus Aedes, A. aegypti and A. albopictus act as vectors for numerous viral infections. Because of non-availability of vaccine for dengue fever, vector control stands the only approach to prevent the viral transmission. As per recommendations by World Health Organization (2015), the main insecticides used for mosquito control are DDT, malathion, chlorpyrifos, temephos, bendiocarb and synthetic pyrethroids. Excessive and unwanted usage of insecticides not only increases vectors’ resistance to insecticides, but also results in cross resistance to other insecticides. In mosquitoes, metabolic detoxifications of insecticides by detoxifying enzymes are the main strategy to withstand recurrent exposure to synthetic insecticides. Esterases are one of the three gene families of detoxification enzymes involved in metabolic detoxification of insecticide and confers resistance for organophosphorus insecticide (OP) as well as for carbamate insecticide. In the present study, the activity of α- and β-esterases have been evaluated in different A. aegypti populations collected from the Dooars and Terai regions of West Bengal, India. The activity of α-esterases was 1.2–3.1 fold and β-esterases was 2.0–23.0 fold higher than laboratory control population. The electro-phoregrams of α-esterases showed expression of 11 isozymes from two gene loci and for β-esterases showed expression of 09 isozymes from two gene loci in all the field populations of A. aegypti. The results showed a significant variation of esterase activity and isozyme expression among different population indicating variability in biochemical susceptibility among the populations. The present work presents first hand data on biochemical resistance of A. aegypti in this region and may be used as an integral component for planning and evaluation of vector-borne diseases and integrated vector management programmes.

Seasonal incidence and enzyme-based susceptibility to synthetic insecticides in two upcoming sucking insect pests of tea

Climate change, deforestation and over reliance on chemical pesticides during the last five decades are presumed to have had a significant impact on the incidence and abundance of agricultural pests. The thrips (Scirtothrips dorsalis Hood) and the greenfly (Empoasca flavescens Fabricius), which were previously considered as minor or occasionally as serious pests in localized areas of tea plantations, are now established as regular and at times major pests in tea plantations of North Bengal spread over the sub Himalayan slopes and the adjoining plains of Terai and the Dooars. Tea-planters from different pockets of North Bengal have been repeatedly reporting control failures of these pests with the use of insecticides in recommended dilutions. A study of their seasonal incidence along with activity of detoxifying enzymes was considered. The trend of a recent population study revealed that E. flavescens and S. dorsalis were abundant throughout the year, with peaks during April to June. Abiotic factors such as temperature, rainfall, sunshine hours and relative humidity (afternoon) influenced the population abundance. Insecticide-exposed populations showed higher levels of activity of the detoxifying enzymes than those collected from organic plantations and the laboratory-reared populations that had not been exposed to insecticides. Detoxifying enzymes in insecticide-exposed E. flavescens collected from Terai and the Dooars as compared with laboratory control specimens showed 3.0–5.2 and 3.0–9.7-fold increases in general esterases (GEs), 1.5–4.8 and 3.6–5.3-fold increases in cytochrome P450s (CYPs) and 1.2–3.5 and 1.5–2.5-fold increases in glutathione-s-transferases (GSTs), respectively. Similarly, S. dorsalis collected from Terai and the Dooars showed 2.0–6.0 and 2.3–5.6-fold increases in GEs, 1.5–2.3 and 1.6–2.4-fold increases in CYPs and 2.6–3.7 and 2.3–3.6-fold increases for GSTs, respectively. Bands of isozymes of esterase I–VI and I–V were found to be expressed in insecticide-exposed specimens whereas a negligible expression was evident for the bands in unexposed E. flavescens and S. dorsalis of organic plantations and laboratory-reared origins, respectively. The data obtained in the present study would be useful in developing the integrated resistance management strategies leading to effective management of the said pests.

Multiple insecticide resistance mechanisms in primary dengue vector, Aedes aegypti (Linn.) from dengue endemic districts of sub-Himalayan West Bengal, India

Background Mosquitoes belonging to genus Aedes are the prime vectors of several arboviral diseases such as Dengue, Zika and Chikungunya worldwide. Every year numerous cases of dengue infections occur throughout the world, proper control of which depends on efficient vector control. However the onset of insecticide resistance has resulted in failure of vector control approaches. Principal findings This study was carried out to unveil the degree of prevailing insecticide resistance along with its underlying mechanisms among the primary dengue vector in dengue endemic districts of West Bengal, India through standard WHO protocol. It was observed that, the majority of the tested populations were found to possess resistance to more than one insecticide. In adult bioassay, the toxicity levels of the six tested insecticides was found to decrease in the following order: deltamethrin > lambdacyhalothrin > malathion > propoxur > permethrin > DDT. In larval bioassay, one of the tested populations was found to possess moderate resistance against temephos, mortality percentage 92.5% and 79.8% for WHO (0.0200 ppm) and National Vector Borne disease Programme, India recommended dose (0.0125 ppm) respectively. Carboxylesterases were found to be involved in conferring resistance as revealed in synergistic and quantitative assay against temephos in North Dinajpur (NDP) population and malathion in Alipurduar (APD) and Darjeeling (DAR) populations. Similar correlations were also observed in the majority of the tested populations between reduced susceptibilities against pyrethroid insecticides and Cytochrome P450s activity. Conclusion Efficient disease management in this region can only be achieved through proper integrated vector management along with tools to minimize insecticide resistance. This study may help the concerned authorities in the formulation of an effective vector control strategy throughout this region incorporating the knowledge gained through this study.

Assessment of insecticide resistance in primary dengue vector, Aedes aegypti (Linn.) from Northern Districts of West Bengal, India

Aedes mosquitoes are the major vectors transmitting several arboviral diseases such as dengue, zika and chikungunya worldwide. Northern districts of West Bengal is home to several epidemics vectored by mosquito including dengue infections, proper control of which depends on efficient vector control. However the onset of insecticide resistance has resulted in failure of vector control approaches. This study was carried out to unveil the level of insecticide resistance prevailing among the primary dengue vector in this dengue endemic region of India. It was observed that, field caught populations of Ae. aegypti were moderately to severely resistant to majority of the insecticide classes tested, i.e. Organochlorine (DDT), Organophosphates (temephos, malathion), Synthetic Pyrethroids (deltamethrin, lambdacyhalothrin and permethrin) and carbamate (propoxur). In majority of the populations, metabolic detoxification seemed to play the underlying role behind the development of insecticide resistance. This study seems to be the first report revealing the pattern of insecticide resistance in Ae. aegypti from Northern West Bengal. Efficient disease management in this region can only be achieved through proper insecticide resistance management. This study may help the concerned authorities in the formulation of an effective vector control strategy throughout this region incorporating the knowledge gained through this study.

Host plant-based variation in fitness traits and major detoxifying enzymes activity in Scirtothrips dorsalis (Thysanoptera: Thripidae), an emerging sucking pest of tea

Scirtothrips dorsalis Hood is a polyphagous species and an important sucking pest of tea (Camellia sinensis) (Theaceae). The fitness traits of S. dorsalis on two alternative host plants: Capsicum annuum L. (chilli) (Solanaceae) and Ricinus communis (castor oil plant) (Euphorbiaceae) and on C. sinensis and corresponding levels of defence enzymes was studied. The study revealed that C. sinensis is the more suitable host of S. dorsalis based on faster development (13.6 days) compared to the alternative hosts, C. annuum (15.5 days) and R. communis (16.7 days), a higher fecundity (C. sinensis: 11.4 eggs; C. annuum: 9.7 eggs; R. communis: 8.6 eggs), and superior egg hatchability (C. sinensis, 92.6%; C. annuum: 82.5%; and R. communis: 74.6%). The host-based variation in the fitness traits of S. dorsalis corroborated in light of differential activity of three major detoxifying enzymes, such as the general esterases (GEs), glutathione S-transferases (GSTs), and cytochrome P450 mediated monooxygenases (CYPs). Densitometric analysis of GEs showed five esterase isozymes (EST I-V) with retardation factor (Rf) values of 0.17, 0.22, 0.27, 0.35 and 0.52, respectively. The pixel density, and accordingly the profile height, varied in different host-specific S. dorsalis. A significant variation of the quantity of these enzymes was also apparent in the insect when reared on the three hosts. A 2.4 and 2.7,1.6 and 2.0, and 2.0 and 2.3-fold higher GEs, GSTs and CYPs activity on the two non-tea hosts possibly signify a predisposition of the species for higher tolerance to insecticides, enabling the pest to switch to tea where synthetic insecticides are routinely used.