Indra Baruah

Insecticide resistance and detoxifying enzyme activity in the principal bancroftian filariasis vector, Culex quinquefasciatus, in northeastern India

The insecticide resistance status of Culex quinquefasciatus Say (Diptera: Culicidae) to DDT and deltamethrin across army cantonments and neighbouring villages in northeastern India was investigated. In India, DDT is still the insecticide of choice for public health programmes. In military stations, pyrethroids, especially deltamethrins, are used for insecticide‐treated nets (ITNs). Recent information on the levels of resistance to DDT and deltamethrin in mosquito populations of northeastern India is scare. Continued monitoring of insecticide resistance status, identification of the underlying mechanisms of resistance in local mosquito populations and the establishment of a baseline data bank of this information are of prime importance. Insecticide susceptibility assays were performed on wild‐caught adult female Cx. quinquefasciatus mosquitoes to the discriminating doses recommended by the World Health Organisation (WHO) to DDT (4%) and deltamethrin (0.05%). Across all study sites, mortality as a result of DDT varied from 11.9 to 50.0%, as compared with 91.2% in the susceptible laboratory strain (S‐Lab), indicating that Cx. quinquefasciatus is resistant to DDT. The species was found to be 100% susceptible to deltamethrin in all study sites except Benganajuli and Rikamari. Knock‐down times (KDT) in response to deltamethrin varied significantly between study sites (P < 0.01) from 8.3 to 17.8 min for KDT50 and 37.4 to 69.5 min for KDT90. All populations exceeded the threshold level of alpha‐esterase, beta‐esterase and glutathion S‐transferase (GST) established for the S‐Lab susceptible strain, and all populations had 100% elevated esterase and GST activity, except Missamari and Solmara. Beta‐esterase activity in Field Unit II (96.9%) was less than in any of the other populations. Benganajuli had the highest activity level for all the enzymes tested. There was a significant correlation between all enzyme activity levels and insecticide resistance phenotype by populations (P < 0.05). The results presented here provide the first report and baseline information of the insecticide resistance status of Cx. quinquefasciatus in northeastern India, and associated information about biochemical mechanisms that are essential for monitoring the development of insecticide resistance in the area.

Molecular analysis of knock down resistance (kdr) mutation and distribution of kdr genotypes in a wild population of Culex quinquefasciatus from India

Objective  To investigate the presence of knock down resistance (kdr) mutation, its frequency distribution in the principal vector of bancroftian filariasis, Culex quinquefasciatus from northeastern India, and to relate kdr genotypes with susceptibility and/or resistance to DDT and deltamethrin in this vectors.

Malaria risk areas of Udalguri district of Assam, India: a GIS-based study

Malaria is a major health problem in India especially in the north-east region. Various methods to combat malaria have been adopted in this region but this problem is yet not under control as all the states report perennial morbidity and mortality due to malaria. The control programmes jeopardized due to improper implementation, inadequate surveillance and lack of geo-referenced information to pinpoint the trouble spots for timely preventive actions. In this investigation an information management system has been constructed based on geographical information system (GIS) for Udalguri district using sub-centre wise malaria data for quick retrieval of information and generation of maps which highlight malaria hot spots. Out of a total of 144 sub-centres, GIS identified 11 sub-centres as malaria hot spots based on annual parasitic incidence >5 and Plasmodium falciparum >30% consistently for 3 years (2006–2008). The district health authorities were advised to focus malaria control in these hot pockets on a priority basis. GIS mapping enables easy update of information which permits policymakers to formulate focused and cost-effective malaria control strategy for endemic areas.

Prioritizing areas for malaria control using geographical information system in Sonitpur district, Assam, India

OBJECTIVE To identify the malaria hot spots at health subcentre level in an endemic district using a geographical information system (GIS). The results will be useful for rapid retrieval of malaria information, and to prioritize malaria control efforts in identified hot spots. STUDY DESIGN Extraction, analysis and synthesis of relevant data. METHODS Malaria epidemiological data from 2006 to 2009 were analysed to determine the annual parasitic index, slide positivity rate, annual blood examination rate and Plasmodium falciparum percentage for each health subcentre in the district. Maps were produced using GIS, and integrated to identify the malaria hotspots. RESULTS Out of 288 health subcentres, GIS identified 10 hot spots at extremely high risk of malaria and 14 hot spots at high risk of malaria. Malaria may flare up in these hot spots whenever favourable transmission conditions arise. Health authorities have been advised to establish control measures in these selected hot spots for timely prevention. CONCLUSION There is a need for adequate monitoring and allocation of available resources for better interventions in the malaria hotspots. The GIS model used in this study can be used, even at village or cluster level, to pin point the malaria hot spots, and information can be updated and retrieved easily.

Insecticide susceptibility and dengue vector status of wild Stegomyia albopicta in a strategically important area of Assam, India

BackgroundDengue vector control programmes are facing operational challenges due to resistance against commonly used insecticides throughout the endemic countries. Recently, there has been appreciable increase in the dengue cases in India, however, no recent data are available on susceptible status of dengue vectors. We have studied the susceptibility level of St. albopicta to commonly used insecticides in India. Adult mosquitoes were tested for the presence of dengue virus.MethodsSt. albopicta larval bioassays were carried out to determine the lethal concentrations (LC10, LC50 and LC99) and the resistance ratios (RR10, RR50 and RR99) for temephos. Susceptibility to 4% DDT, 0.05% deltamethrin and 5% malathion was assessed following standard procedure. Knock-down times (KDT10, KDT50 and KDT99) were estimated and knock-down resistance ratios (KRR10, KRR50 and KRR99) were calculated. VectorTest™ dengue antigen assay was used to detect the dengue virus in the field collected mosquitoes.ResultsIn larval bioassays, the RR ranged from 1.4 (for RR99) to 1.7 (for RR50), which suggested that the tested St. albopicta were susceptible to temephos. There was no deviation among the lethal concentration data from linearity (r2 = 0.61). Adult St. albopicta mosquitoes were resistant to DDT, while fully susceptible to deltamethrin and malathion. The knock-down values (KDT10, KDT50 and KDT99) obtained for DDT displayed straight line in log-dose-probit analysis and follow linear regression model. The KRR99 for DDT was 4.9, which indicated a 4.9 folds increase in knock-down resistance to DDT. However, for malathion and deltamethrin, the KRR99 values were 1.6 and 1.5 respectively suggesting that mosquitoes were knock-down sensitive. None of the mosquito pool was dengue virus positive.ConclusionSt. albopicta showed resistance to DDT and reduced sensitivity to deltamethrin and malathion. This data on insecticide resistance could help public health authorities in India to design more effective vector control measures. More dengue vector specimens need to be scanned to identify the potential dengue vector.

Polymerase chain reaction detection of human host preference and Plasmodium parasite infections in field collected potential malaria vectors.

Abstract This study was carried out to determine the human host preference and presence of Plasmodium parasite in field collected Anopheles mosquitoes among four villages around a military cantonment located in malaria endemic Sonitpur district of Assam, India. Encountered malaria vector mosquitoes were identified and tested for host preference and Plasmodium presence using PCR method. Human host preference was detected using simple PCR, whereas vectorial status for Plasmodium parasite was confirmed using first round PCR with genus specific primers and thereafter nested PCR with three Plasmodium species specific primers. Out of 1874 blood fed vector mosquitoes collected, 187 (10%) were processed for PCR, which revealed that 40·6% had fed on human blood; 9·2% of human blood fed mosquito were harbouring Plasmodium parasites, 71·4% of which were confirmed to Plasmodium falciparum. In addition to An. minimus, An. annularis and An. culicifacies were also found positive for malaria parasites. The present study exhibits the human feeding tendency of Anopheles vectors highlighting their malaria parasite transmission potential. The present study may serve as a model for understanding the human host preference of malaria vectors and detection of malaria parasite inside the anopheline vector mosquitoes in order to update their vectorial status for estimating the possible role of these mosquitoes in malaria transmission. The study has used PCR method and suggests that PCR-based method should be used in this entire malarious region to correctly report the vectorial position of different malaria vectors.

High-throughput approach to detection of knockdown resistance (kdr) mutation in mosquitoes, Culex quinquefasciatus, based on real-time PCR using single-labelled hybridisation probe/melting curve analysis.

BACKGROUND Knockdown resistance (kdr) mutation (L1014F) is a well-defined mechanism of resistance to pyrethroids and DDT in many insect species. Sensitive detection of the mutations associated with resistance is a prerequisite for resistance management strategies. The authors have developed a new real-time molecular diagnostic assay based on SimpleProbe(®)/melting curve analysis for large-scale kdr genotyping in the wild population of Culex quinquefasciatus Say, the principal vector of bancroftian filariasis. Melting curve analysis is based on the thermal stability difference between matched and mismatched DNA duplexes. The application of SimpleProbe(®) chemistry in insects described here is novel in entomology research. RESULTS The mosquitoes homozygous for knockdown-resistant and knockdown-susceptible allele showed melting peaks at 60.45 °C (±0.25) and 64.09 °C (±0.24) respectively. The heterozygous mosquitoes yielded both peaks at approximately 60.5 °C (±0.2) and 64.20 °C (±0.23). Among the 92 samples genotyped, 16 were found to be homozygous resistant, 44 homozygous susceptible and 32 heterozygous. Comparative assessments were made of all the reported methods for kdr genotyping. CONCLUSION The present method is cheaper, faster, more reliable and versatile than other alternatives proposed in detecting correct kdr genotypes in mosquitoes. This is the first report using a single-labelled hybridisation probe to detect point mutations in insect populations.

A longitudinal study of malaria associated with deforestation in Sonitpur district of Assam, India

Assam–Arunachal forest fringed foothill area is endemic for malaria incidence. The present study deals with the temporal analysis of malaria incidence and determines its association with deforestation in 24 villages along the Assam–Arunachal forest fringed foothill area of Sonitpur district of Assam. Malaria epidemiological survey has been carried out in the study area from the year 1994 to 2005. Remote sensing (RS) technique has been used to map the areas of forest changes from the year 2000 to 2005. Geographical information system (GIS) was used to map the malaria incidence and forest cover. The study villages are endemic to malaria infections and there was increasing trend of malaria incidence over the years. The slide positivity rate (SPR) ranged from 5.1% in 1997 to 44.4% in 2005. The percentage forest cover decreased significantly from 23.6% during 2000 to 15.4% during 2005, whereas SPR was increased during 2000–2005. The present study is the first attempt to understand the role of deforestation in malaria incidence using RS and GIS in the north-eastern region of India at a micro-geographic level. The study suggests that the area is endemic to malaria transmission. The decrease in forest cover is a serious ecological concern besides its role in elevating the malaria incidence in the study area.

Chemotherapy and drug resistance status of malaria parasite in northeast India

India reports the highest number of malaria cases in Southeast Asia, of which Plasmodium falciparum contribute more than half of the cases every year. North eastern states of India contribute only 3.96% of countrys population but account for >10% of total reported malaria cases, 11% of Plasmodium falciparum cases and 20% of malaria related deaths annually. In India, chloroquine resistance was reported for the first time from northeast region and since then chloroquine treatment failure is being reported from many parts of the region. Increased chloroquine treatment failure has led to change of the drug policy to artemisinin combination therapy as first line of malaria treatment in the region. However, replacing chloroquine to artemisinin combination therapy has not shown significant difference in the overall malaria incidence in the region. The present review addresses the current malaria situation of northeastern region of India in the light of antimalarials drug resistance.

Development of physiological resistance and its stage specificity in Culex quinquefasciatus after selection with deltamethrin in Assam, India

The study investigated the development and stage specificity of physiological resistance to insecticides in a colony of Culex quinquefasciatus Say (Diptera: Culicidae) mosquitoes, which are vectors of bancroftian filariasis in India, after selection with deltamethrin. Resistance was selected by exposing the larvae to the concentration of deltamethrin that caused 50% mortality in the tested population (i.e., LC50). Under continuous selection pressure, the LC50 increased steadily in subsequent generations. The estimated LC50 for the F0 generation was 0.409 microg/L; the LC50 first displayed a substantial increase in the F5 generation (5.616 microg/L) and reached 121.902 microg/L in the F10 generation. The objective of this study was to establish a deltamethrin-resistant colony to develop a research programme that will study the evolution of physiological resistance patterns and stage-specific resistance responses in Cx. quinquefasciatus larvae and adults under laboratory conditions. An approximately 298-fold increase in resistance was recorded after 10 generations, as evidenced by the resistance ratio (RR50). The progress and effect of the selection pressure in the adult stage was monitored with the World Health Organisation (WHO) diagnostic test. The mortality, as observed using the WHO diagnostic test, declined significantly from the F5 generation (85%) onwards and the highest rate of survival (65%) was observed in the F10 generation.