Finishing the task of eliminating neglected tropical diseases (NTDs) in WHO South-East Asia Region: promises kept, challenges, and the way forward.

Globally 20 Neglected tropical diseases (NTDs) are prioritized by World Health Organization (WHO), of which 15 are present in the South-East Asia Region (SEAR) with all 11 countries being affected. As the region bears 54% of the global burden, "Finishing the task of eliminating neglected tropical diseases and other diseases on the verge of elimination" was identified as a regional flagship priority in 2014 with focus on lymphatic filariasis (LF), kala-azar, yaws, trachoma, and leprosy. Intensified efforts have been made to raise and sustain political commitment and momentum among partners innovate tools, interventions and strategies to accelerate elimination, and establish the process and support countries to accelerate and validate achievement of elimination targets. Seven countries have verified or validated for having eliminated at least one NTD since 2016, including yaws, LF and trachoma. Between 2010 and 2020, the number of people requiring interventions against NTDs in the South-East Asia Region reduced by 20%. The priorities in the next decade are to strengthen last-mile efforts to eliminate identified NTDs, sustain it and to use the lessons learnt to eliminate other NTDs. Funding: None.

Recent advances on T-cell exhaustion in malaria infection.

T-cell exhaustion reportedly leads to dysfunctional immune responses of antigen-specific T cells. Investigations have revealed that T cells expand into functionally defective phenotypes with poor recall/memory abilities to parasitic antigens. The exploitation of co-inhibitory pathways represent a highly viable area of translational research that has very well been utilized against certain cancerous conditions. Malaria, at times, evolve into a sustained chronic state where T cells express several co-inhibitory molecules (negative immune checkpoints) facilitating parasite escape and sub-optimal protective responses. Experimental evidence suggests that blockade of co-inhibitory molecules on T cells in malaria could result in the sustenance of protective responses together with dramatic parasite clearance. The role of several co-inhibitory molecules in malaria infection largely remain unclear, and here we discussed the potential applicability of co-inhibitory molecules in the management of malaria with a view to harness protective host responses against chronic disease and associated consequences.

Control of Aedes aegypti and Ae. albopictus, the vectors of dengue and chikungunya, by using pheromone C21 with an insect growth regulator: Results of multicentric trials from 2007-12 in India.

BACKGROUND & OBJECTIVES: Aedes mosquito control has gained much importance nowadays in view of rise in number of reported cases of dengue and chikungunya in India and other countries. In the present study, C21 attracticide (containing a pheromone and an insect growth regulator­IGR, developed by Defence Research and Development Establishment (DRDE), Gwalior, India was tested for its feasibility for surveillance and control of Aedes mosquito in a multicentric mode from October 2007 to June 2012 in urban (Delhi, and Bengaluru district, Karnataka) and suburban (Alappuzha district, Kerala) settings of the country in three phases. METHODS: Across the randomly selected households in each study area, two to four containers treated with attracticide (experimental) and untreated (control) were placed and monitored by trained surveillance workers on weekly/ fortnightly basis for determining the presence of eggs, larvae and pupae. Container positivity, percent larvae, egg and pupae collected were determined during different phases and analyzed statistically using SPSS 18.0. RESULTS: Container positivity was found statistically significant at Bengaluru and Alappuzha, Kerala while in Delhi, it was found non-significant. Eggs collected from experimental containers were significantly higher in comparison to control at all the locations except Delhi. Also larvae collected from control containers were significantly higher at all the locations except Bengaluru. Pupae collected from control containers remained significantly higher at all the locations as no pupal formation was recorded from experimental containers. INTERPRETATION & CONCLUSION: The use of C21 attracticide hampered pupal formation, thus inhibiting adult population in the study areas. The study established that C21 attracticide was efficacious in the field conditions and has potential for use in surveillance and management of dengue and chikungunya mosquitoes.

A note on the insecticide susceptibility status of principal malaria vector Anopheles culicifacies in four states of India.

BACKGROUND & OBJECTIVES: The major malaria vector, Anopheles culicifacies Giles is reported to contribute ~ 65% of the malaria cases in India. This species developed resistance to DDT and later to HCH, malathion and also to pyrethroids in some states due to their use in the national malaria control programme. In the present study, insecticide susceptibility of this species was monitored in four states of India. METHODS: To determine insecticide susceptibility status of the major malaria vector An. culicifacies, adult mosquitoes were collected from different localities of 32 tribal districts in the states of Andhra Pradesh, Odisha, Jharkhand and West Bengal during October/November 2009-10. Mosquitoes were collected from stratified ecotypes comprising a group of districts in West Bengal and individual districts in three other states. Mosquitoes were exposed to papers treated with WHO diagnostic dose: 4% DDT, 5% malathion and 0.05% deltamethrin following the WHO tube method. RESULTS: RESULTS provided the susceptibility status of An. culicifacies to different insecticides used in the public health programme in 32 districts in four states. An. culicifacies was found resistant to DDT (mortality range 0-36%) in all the 32 districts; to malathion it was resistant in 14 districts, verification required in 10 districts and susceptible in eight districts (mortality range 32.2-100%). It was resistant to deltamethrin in four districts, verification required in 11 districts and susceptible in 17 districts (mortality range 43.3-100%). INTERPRETATION & CONCLUSION: Development of widespread resistance to insecticides used in public health sprays for vector control including to pyrethroids in An. culicifacies in the surveyed districts is of great concern for the malaria control programme as the major interventions for vector control are heavily reliant on chemical insecticides, mainly synthetic pyrethroids used both for indoor residual spraying and for long-lasting insecticidal nets. Thus, there is a need to periodically monitor and update the susceptibility status of malaria vector(s) to suggest alternative vector control strategies for effective disease management.

Emerging and re-emerging arboviral diseases in Southeast Asia.

Arthropod-borne viruses (arboviruses) have become significant public health problems, with the emergence and re-emergence of arboviral diseases nearly worldwide. The most populated Southeast Asia region is particularly vulnerable. The arboviral diseases such as dengue (DEN), Japanese encephalitis (JE), West Nile virus (WNV), chikungunya fever (CHIK), hemorrhagic fevers such as Crimean-Congo hemorrhagic (CCHF) fever, Kyasanur forest disease virus (KFDV), etc. are on the rise and have spread unprecedentedly, causing considerable burden of disease. The emergence/re-emergence of these diseases is associated with complex factors, such as viral recombination and mutation, leading to more virulent and adaptive strains, urbanization and human activities creating more permissive environment for vector-host interaction, and increased air travel and commerce. Climate is a major factor in determining the geographic and temporal distribution of arthropods, the characteristics of arthropod life cycles, the consequent dispersal patterns of associated arboviruses, the evolution of arboviruses; and the efficiency with which they are transmitted from arthropods to vertebrate hosts. The present and future arboviral threats must be mitigated by priority actions such as improving surveillance and outbreak response, establishing collaboration and communication intersectorally, and strengthening the prevention and control programmes along with improving biosafety aspects with regards to highly infectious nature of these arboviral diseases. Evidence from research needs to be generated and priority areas for research defined.

Safety, efficacy and population pharmacokinetics of fixed-dose combination of artesunate-mefloquine in the treatment of acute uncomplicated Plasmodium falciparum malaria in India.

BACKGROUND & OBJECTIVES: India has switched over to artemisinin-based combination therapy (ACT) for the treatment of acute uncomplicated Plasmodium falciparum malaria and the ACT used in the national programme is artesunate + sulphadoxine-pyrimethamine. Since the efficacy of ACT is dependent also on the partner drug, there is a need to evaluate and deploy multiple ACTs. METHODS: This multicentre, single-arm, open-label clinical trial was carried out to assess the efficacy, safety and population pharmacokinetics of a fixed dose combination (FDC) artesunate mefloquine (ASMQ) in P. falciparum infected, Indian adults at Panjim, Goa, and Mangalore, Karnataka between December 2007 and November 2008. RESULTS: A total of 77 patients (males 74) were screened and enrolled: 42 at Goa and 35 at Mangalore with a median age of 25 yr (range 18-55 yr). One patient failed in treatment on D53, a PCR proven new infection, seven developed recurrent vivax parasitaemia and 11 did not have a parasitological endpoint. By per protocol analysis, the D63 cure rate was 58/59 (98.3; 95% C.I. 90.9-99.9%), and 58/58, with PCR correction. ASMQ was well-tolerated and no serious adverse events were reported. INTERPRETATION & CONCLUSION: The study showed that the ASMQ FDC was efficacious and well-tolerated for the treatment of acute, uncomplicated P. falciparum malaria in highly endemic, chloroquine resistant areas of Goa and Mangalore. It is a viable option for India.

Resting behaviour of Anopheles stephensi type form to assess its amenability to control malaria through indoor residual spray.

BACKGROUND & OBJECTIVES: In the present study, an attempt was made to find reasons of ineffectiveness of indoor residual spray to interrupt malaria transmission by investigating the behaviour of two variants of Anopheles stepnesi, viz. type form and mysorensis in rural areas of Rajasthan, India. METHODS: Both low malarious (DDT spray from 1958 to 1976) and high malarious (DDT spray from 1958 to April 2006) villages were selected for the study in Arid zone district Jodhpur, Rajasthan during March-April 2006. Resting behaviour of the species during all its movement rhythms covering 24 h period related to: (i) swarming/ mating; (ii) pre- and post-biting rest; (iii) after feed resting between hoping movements; (iv) night and day time resting and; (v) diel activity movements in response to temperature changes, were carried out. RESULTS: The results showed that household objects like cupboards, furnitures, hanging clothes, goods stacked on loft, stored clothes, cobwebs and floor were the dominant resting sites both in sprayed and unsprayed villages. About 95 and 97% of An. stephensi preferred to rest on household objects of unsprayed and sprayed villages respectively. There was no significant difference in resting behaviour of the species in both groups of villages (p >0.05). The pre-biting resting time was recorded as 5 to 15 min whereas post-biting resting time lasted for 15 to 25 min. After biting outdoor (in courtyard--open to sky) species starts entering the rooms at around 2330 hrs. It was observed that during III quarter (0100 to 0400 hrs) maximum species entered into the room were 56% in unsprayed and 62% in sprayed villages. Statistically there was no significant difference in the entry of mosquitoes (p >0.05) in both the groups of villages. CONCLUSION: Before DDT era, An. stephensi was found resting at all heights of the walls inside the human dwellings. Present study revealed that An. stephensi is trying to avoid sprayable surfaces and tend to rest on unsprayable surfaces during all its movement rhythms starting from swarming, pre-/post-biting and during hopping movements in the courtyard and thereafter household objects inside the room as final day time resting. It was concluded that changed behaviour of resting of An. stephensi on unsprayable objects in sprayed rooms largely accounted for failure of malaria control. Control of An. stephensi, thus requires an integrated vector control strategy based upon interesectoral, environmental, larviciding with chemical/biolarvicide and use of larvivorous fish wherever feasible. Such a control strategy offers cost-effective and sustainable option than indoor residual spray.

Field evaluation of Icon®Life, a long-lasting insecticidal net (LLIN) against Anopheles culicifacies and transmission of malaria in District Gautam Budh Nagar (Uttar Pradesh), India.

BACKGROUND & OBJECTIVES: In the present study, Icon®Life net, a long-lasting polyethylene net, 100 denier and bursting strength of minimum 280 kpa incorporated with deltamethrin @ 65 mg/m2 was evaluated for its efficacy in reducing the density of malaria vector Anopheles culicifacies and impact on malaria prevalence in a malaria endemic area of District Gautam Budh Nagar, India. METHODS: Wash resistance of Icon®Life LLIN was determined up to 20 serial washings using An. culicifacies in cone bioassays under field conditions. Efficacy of Icon®Life LLIN was determined in the field in three sets of villages in District Gautam Budh Nagar (Uttar Pradesh), India, selected randomly for the intervention with Icon®Life LLIN, untreated nets and a control without any intervention for the period of August 2008-July 2009. Entomological and malariometric indices in all the three villages were compared during pre- and post-intervention periods for one year against An. culicifacies. A survey was also conducted in the village provided with Icon®Life LLIN to assessing the perception of community regarding acceptance of these nets by the community. RESULTS: In cone bioassays on Icon®Life LLIN with An. culicifacies, >95% knockdown within 1 h and 100% mortality after 24 h exposure were reported even after 20× serial washings under field conditions. Results of the field study revealed reduced entry rate, resting density and parity rate of An. culicifacies in the village with Icon®Life LLIN when compared to no net and untreated net villages. Number of malaria cases reported were less in the Icon®Life LLIN used villages when compared to other two villages. The community compliance and acceptance was high and no adverse health events were reported by the households using these nets. CONCLUSIONS: Icon®Life LLIN is an effective intervention for the control of An. culicifacies transmitted malaria in India. Long-term studies are indicated for the duration of effectiveness and to ascertain the epidemiological impact of the use of Icon®Life nets.

In vitro assessment of drug resistance in Plasmodium falciparum in five States of India.

BACKGROUND & OBJECTIVES: In vitro assays are an important tool to assess baseline sensitivity and monitor the drug response of Plasmodium falciparum over time and place and, therefore, can provide background information for the development and evaluation of drug policies. This study was aimed at determining the in vitro sensitivity of P. falciparum isolates to antimalarials. METHODS: The in vitro activity of 108 P. falciparum isolates obtained from five States of India was evaluated using WHO microtest (Mark III) to chloroquine, monodesethylamodiaquine, dihydroartesunate and mefloquine. Samples were collected from the States of Orissa, Jharkhand, Karnataka, Goa and Chhattisgarh from September 2007 to August 2009. In addition, representative samples from different States of India cryopreserved and culture adapted in the Malaria Parasite Bank of National Institute of Malaria Research, New Delhi, were also evaluated. RESULTS: The proportion of isolates resistant to chloroquine and monodesethylamodiaquine was 44.4 and 25 per cent, respectively. Of the 27 isolates resistant to monodesethylamodiaquine, 16 (59.3%) were cross-resistant to chloroquine. No isolate showed resistance to dihydroartesunate and mefloquine. Isolates from Orissa showed the highest degree of resistance to chloroquine and amodiaquine followed by Jharkhand. Forty two isolates were genotyped for pfcrt T76K chloroquine resistant mutation; mutations were seen in 38 (90.47%) isolates. INTERPRETATION & CONCLUSIONS: The Indian P. falciparum isolates showed a high degree of resistance to chloroquine followed by monodesethylamodiaquine. No resistance was recorded to mefloquine and dihydroartesunate.

Retrospective study of chikungunya outbreak in urban areas of India.

BACKGROUND & OBJECTIVES: A retrospective study on chikungunya outbreak in India in five States viz. Delhi, Madhya Pradesh, Orissa, Maharashtra and Kerala was conducted in 2007-2008 to know the distribution and determinants of chikungunya fever outbreak in India. METHODS: On the basis of high and low incidence of chikungunya fever, two districts from each State and two wards from the selected district were taken for random selection of 1000 households from 10 districts and 5 States. Semi-structured questionnaires were administered to individuals, patients, qualified health professionals and to stakeholders for collecting information. RESULTS: The educational background and occupation of the respondents showed variations across the study States. Only in high incidence ward of Maharashtra, water storage period for 3-6 days and emptying, drying of water containers on weekly basis was noted. The study through knowledge, attitude, belief, practice (KABP) obtained individual's perception of chikungunya fever, its prevention and control. Patients' expenditure on treatment was mainly recorded less than Rs 500 across study States. Health facility survey obtained an overview of the capacity of local health facilities. Stakeholders' perception regarding chikungunya fever was also noted. INTERPRETATION & CONCLUSIONS: The study revealed differences in awareness of chikungunya, cause of the disease, vector responsible, mode of transmission, biting time and elimination of breeding of mosquitoes statistically significant among high and low incidence wards of all the States. Expenditure on treatment was independent of economically active status and loss of man-days across all the States. Education and occupation did not have any relation with emptying/drying of water containers in high incidence wards. Strengthening of surveillance, information, education and communication (IEC) activities along with case management facilities may be provided by the State health department for prevention of chikungunya outbreaks in future. Stakeholders should be more involved in outbreak management and future planning.

Evaluation of toxicity of plant extracts against vector of lymphatic filariasis, Culex quinquefasciatus.

BACKGROUND & OBJECTIVES: Conventional insecticides are generally used as larvicides to control Culex quinquefasciatus, vector of lymphatic filariasis. This study was undertaken to evaluate the larvicidal activity of some potential larvicidal plants leaf extracts against Cx. quinquefasciatus larvae. METHODS: The toxic effects of petroleum ether leaf extracts of plants viz., Argemone mexicana (Mexican prickly poppy), Clausena dentata (Dentate), Cipadessa baccifera (Rana bili), Dodonaea angustifolia (Hop bush) and Melia dubia (Pride of India) were evaluated under laboratory conditions in individual and in combination against 3 rd - 4 th instar larvae of Cx. quinquefasciatus. RESULTS: The results indicated that among the selected plants, A. mexicana showed maximum larvicidal activity with an LC 50 value of 48.89 ppm. Its toxicity was enhanced when the extract was mixed (1:1) with that of C. dentata as the LC 50 value became 28.60 ppm indicating synergistic action of A. mexicana. INTERPRETATION & CONCLUSIONS: Our results showed high larvicidal potential in A. mexicana leaf extract, and it also showed additive effect when mixed with C. dentata extract.

Evaluation of the impact of ZeroFly®, an insecticide incorporated plastic sheeting on malaria incidence in two temporary labour shelters in India.

BACKGROUND & OBJECTIVES: Prevention of malaria is a major technical and operational problem in displaced and mobile populations such as refugee camps and temporary labour settlements. Insecticide incorporated plastic sheeting is a new technology to control mosquitoes in emergency shelters and also temporary habitations at different locations. In view of this, efficacy of ZeroFly® , an insecticide incorporated plastic sheeting (factory treated with deltamethrin 2.0 g/kg or 265 mg/m2 ) was evaluated for its efficacy against malaria vectors and its impact on malaria incidence in temporary labour settlements in two urban areas in India. METHODS: This trial was conducted in two labour settlements in two urban areas, Delhi and Noida (U.P.), India with ~ 250 populations. In an area, two localities were selected for intervention with ZeroFly and untreated plastic sheets (control). Entomological and epidemiological data were collected using standard methods for one year. RESULTS: Baseline studies on the susceptibility of mosquitoes in Delhi and Noida areas revealed 100% susceptibility of the malaria vector species Anopheles culicifacies and An. stephensi to deltamethrin. Cone bioassay tests performed against An. culicifacies and An. stephensi to determine the efficacy of ZeroFly sheets showed 100% mortality against An. culicifacies and An. stephensi with 3 min exposure and after 24 h recovery period. Against Culex quinquefasciatus and housefly 100% mortality was obtained after 30 min of exposure period. Intervention with the ZeroFly plastic sheets resulted in almost complete reduction in the resting density of An. culicifacies and An. stephensi, the two major malaria vectors and also in the reduction of malaria cases in ZeroFly camps as compared to control camps. The ZeroFly plastic sheeting was found to be safe for human. Barring some complaints of skin irritation and itching, which were temporary in nature, no adverse health effects were reported by the users. The community acceptance was high. CONCLUSION: Results of the present study revealed that ZeroFly® plastic sheeting is highly effective in reducing the indoor resting density of mosquitoes, man -vector contact and malaria incidences in labour populations living in temporary shelters.

Efficacy of Advanced Odomos repellent cream (N, N-diethyl-benzamide) against mosquito vectors.

BACKGROUND & OBJECTIVES: Repellents are commonly used personal protection measures to avoid mosquito bites. In the present study, Advanced Odomos cream (12% N, N-diethyl-benzamide) was tested for its efficacy against mosquitoes in comparison to DEET (N,N-diethyl-3-methyl benzamide). METHODS: Bioassays were conducted to assess the repellency of Advanced Odomos and DEET creams against Anopheles stephensi and Aedes aegypti. Their efficacy was tested on human volunteers applied with different concentrations of test creams ranging from 1 to 12 mg/cm 2 and by exposing them to mosquitoes at hourly intervals. Field evaluation was also carried out to test the duration of protection of the test creams against Anopheles and Aedes mosquitoes during whole night and day time collections, respectively on human volunteers. Mosquito collections were done using torch light and aspirator. RESULTS: Complete (100%) protection was achieved at 10 mg/cm 2 cream formulation of Advanced Odomos (1.2 mg a.i/cm 2 ) dose against An. stephensi and 12 mg/cm 2 (1.44 mg a.i./cm 2 ) against Ae. aegypti on human baits. There was no statistically significant differences in per cent protection against mosquito bites between Advanced Odomos and DEET cream (P>0.05) in respective doses. Complete protection up to 11 h was observed against Anopheles mosquitoes during whole night collections and up to 6 h against Ae. aegypti in day time collections. No adverse reactions such as itching, irritation, vomiting, nausea, etc. were reported by the volunteers. INTERPRETATION & CONCLUSIONS: Advanced Odomos cream applied at 10 mg/cm 2 concentration provided 100% protection from Anopheles mosquitoes up to 11 h whereas about 6 h protection was recorded against Ae. aegypti. The laboratory and field trials indicate that for longer protection against Anopheles mosquitoes 10 mg/cm 2 will be appropriate and in case of Ae. aegypti more than 10 mg/cm 2 application is required for complete protection. In conclusion, the Advanced Odomos cream was comparable to the known repellent cream DEET for prolonged protection against malaria and dengue vectors.

Field evaluation of lambda-cyhalothrin (ICON 10 CS) indoor residual spraying against Anopheles culicifacies in India.

BACKGROUND & OBJECTIVES: Field trials of lambda-cyhalothrin 10 CS (ICON 10 CS) in indoor residual spraying (IRS) with 25 mg a.i./m2 against Anopheles culicifacies was undertaken vs malathion IRS (25% WP-2 g a.i./m2) in Tumkur district, Karnataka; vs deltamethrin IRS (2.5% WP-20 mg a.i./m2) in Dharmapuri district; and vs lambda-cyhalothrin (10 WP-25 mg a.i./m2) in Ramanathapuram district, Tamil Nadu, India. METHODS: Spray operations in the experimental villages were done by the National Institute of Malaria Research (NIMR) and in the control villages by the respective State Health Department staff. Persistence of efficacy of insecticide sprayed in villages was assessed by contact bioassays against vector mosquitoes. Entomological indicators such as per structure density, parity rates of vector mosquitoes and sporozoite rates were measured in all the three study areas using standard procedures. Mass blood surveys and active fever case detections were carried out in experimental and control villages to study the impact of IRS on malaria transmission. RESULTS: Persistence of effectiveness of ICON 10 CS was observed up to 2-3 months in all the three study areas. ICON 10 CS was found effective at par with or better than the insecticides used in the national programme in reducing the mosquito densities and in interrupting malaria transmission in the study villages. Vector density, parity rates and malaria cases considerably reduced in the ICON 10 CS-sprayed villages. CONCLUSION: Field trials at three sites have established that ICON 10 CS formulation was relatively more effective than malathion 25% WP, deltamethrin 2.5% WP and lambda-cyhalothrin 10% WP in some evaluation parameters like indoor resting mosquitoes, parity rates in vector mosquitoes and persistence of effectiveness. It can be used for IRS for malaria vector control with two rounds of spray at an interval of 3 months for curtailing the malaria transmission and an additional round is recommended in perennial malaria transmission areas.

Evaluation of the pyrrole insecticide chlorfenapyr for the control of Culex quinquefasciatus Say.

Culex quinquefasciatus Say (Diptera: Culicidae) is a widely distributed mosquito vector species in India and also in other tropical regions of the world. This species is implicated in the transmission of lymphatic filariasis in many countries. This species is reported to be widely resistant to insecticides of different classes in current use. In the present study, bio-efficacy of chlorfenapyr, an insecticide of pyrrole class with a novel mode of action was tested for the control of Cx. quinquefasciatus. Studies were performed to determine the diagnostic dosage; residual efficacy on different artificially fabricated substrates, namely wood, mud, mud+lime, cement and cement+distemper; to assess cross-resistance with different insecticides; and synergism/antagonism using piperonyl butoxide (PBO). A dosage of 5.0% chlorfenapyr was determined as diagnostic dosage with 2 h exposure and 48 h holding period for assessing the susceptibility of mosquitoes. The residual efficacy was observed up to 34 weeks on wood and mud+lime substrates while on other substrates, it was about 15 weeks at a dosage of 400mg a.i./m(2). Laboratory-reared strains of Cx. quinquefasciatus showed cross-resistance, whereas field-collected mosquitoes showed absence of cross-resistance to chlorfenapyr. Potentiation bioassays showed antagonistic effect of PBO to chlorfenapyr toxicity owing to the involvement of oxidases in the initial step of a conversion of pro-insecticide chlorfenapyr to toxic form CL 303268. The present study results have shown that chlorfenapyr can be a potential insecticide for the control of multiple insecticide resistant strains of Cx. quinquefasciatus. However, in countries where indoor residual spray (IRS) is not targeted for the control of this species, like in India, chlorfenapyr used in IRS for the control of malaria vectors in rural and peri-urban areas can additionally provide control of Cx. quinquefasciatus also.

Molecular differentiation of nocturnally periodic and diurnally sub-periodic Wuchereria bancrofti by Randomly Amplified Polymorphic DNA (RAPD).

Wuchereria bancrofti, a nematode parasite, is responsible for causing 90% of lymphatic filariasis infection in the world. In India, W. bancrofti exists in two physiological forms, nocturnally periodic in the main land and diurnally sub-periodic in Car Nicobar group of islands. Differentiation of these two parasitic forms by conventional microscopic methods difficult requiring good skill and hence tedious. Therefore, we developed a simple and rapid Random Amplified Polymorphic DNA (RAPD) assay to differentiate these parasitic forms. Also, the phylogenetic relationship between periodic and sub-periodic W. bancrofti and also Brugia malayi populations was analyzed using RAPD profile generated. Distinct RAPD profiles were observed among the three parasites with the formation of three distinct clusters in the phylogenetic tree. Substantial genetic diversity (Nei's genetic diversity H) was observed among periodic (H = 0.0577) and sub-periodic (H = 0.1415) W. bancrofti populations.

Wash resistance and efficacy of Olyset net and Permanet 2.0 against Anopheles stephensi in India.

Long-lasting insecticidal nets (LLIN) have been developed for wash resistance and long-lasting effects against mosquito vectors. In this study we evaluated 2 LLIN products, Olyset net and Permanet 2.0, washed for 0, 5, 10, 15, and 20 times, against Anopheles stephensi, an urban malaria vector in India. We assessed the wash resistance and efficacy of these nets in relation to bloodfeeding inhibition and percent mortality in cone and tunnel test bioassays. Both Olyset and Permanet showed >80% mortality of An. stephensi in cone bioassays after 20 washes. In tunnel tests there was no significant difference between Olyset and Permanet 2.0 in causing total mosquito mortality (immediate and delayed) up to 10 washes and bloodfeeding inhibition and entry rate up to 15 washes. After the 20th wash, Permanet 2.0 was significantly more effective than the Olyset net in causing total mosquito mortality, whereas Olyset net showed less bloodfeeding and entry of mosquitoes as compared to Permanet 2.0. There was a gradual decline in efficacy of both LLIN products as the number of washings increased. Cone bioassays indicated a strong wash resistance in both the LLIN products after 20 washes. However, the tunnel tests demonstrated a gradual decline in efficacy of both products with the number of washings.

Community awareness, perceptions, acceptability and preferences for using LLIN against malaria in villages of Uttar Pradesh, India.

BACKGROUND & OBJECTIVES: Long-lasting insecticidal nets (LLINs) have been introduced in India recently into the vector control programme. A study was undertaken to assess the community perception regarding use of LLIN, their acceptability, collateral benefits, etc. in certain villages of District Gautam Budh Nagar, Uttar Pradesh, India where LLINs are in use by the community. METHOD: A randomized community-based survey was undertaken during April- May 2009 covering 596 respondents in LLIN villages where LLINs have been distributed along with 307 respondents in untreated net villages where untreated nets were distributed using structured questionnaire. Qualitative data were collected and the difference in proportion was calculated by z-test. RESULTS: A substantial number of respondents had good knowledge of the symptoms of malaria. According to respondents, LLINs were very much effective in bringing down the malaria incidence in their families. About 98.3% of the respondents asserted the use of LLINs as their use not only reduced the number of mosquitoes as well as other non-target insects, but also reduced the malaria incidence. About 93.2% of the respondents were ready to purchase LLINs if available at nominal prices. All the respondents were satisfied about the performance of the LLINs in reducing the mosquito nuisance, safety of use and collateral benefits in LLIN villages. CONCLUSION: LLINs are safe, socially acceptable and should be promoted for vector control to reduce the disease burden in the communities.

Persistence of DDT, malathion & deltamethrin resistance in Anopheles culicifacies after their sequential withdrawal from indoor residual spraying in Surat district, India.

BACKGROUND & OBJECTIVES: Under the national antimalaria programme DDT was introduced in early 1950s for vector control and later hexachloro cyclohexane (HCH) followed by malathion and recently synthetic pyrethroids in 1990s to manage the insecticide resistance in Anopheles culicifacies. Subsequent replacement led to development of multiple resistances in An. culicifacies in Surat district in Gujarat State. Indoor residual spray (IRS) was completely withdrawn in southern villages in Surat in 2002. This study was undertaken in these areas to study the persistence of resistance to DDT, malathion and deltamethrin after sequential withdrawal of IRS with these insecticides at different times. METHODS: Susceptibility tests on An. culicifacies were conducted using standard WHO methods and kits. Mortality, knockdown time and lethal times were calculated for An. culicifacies exposed to WHO prescribed diagnostic concentrations of different insecticide impregnated papers. RESULTS: Persistence of DDT-resistance was observed even after 30 yr of its withdrawal from IRS. Similarly, persistence of malathion resistance was also observed after 9 yr of its withdrawal from IRS, while reversal of deltamethrin-resistance was observed very fast within 2-3 yr after its withdrawal from IRS in 2002. INTERPRETATION & CONCLUSION: Present data indicate that the quantum of reversion of insecticide resistance in a population is relative and depends on the genetic stability of the respective resistance genes in the mosquitoes. In the present study withdrawal of pyrethroid-IRS resulted in increased susceptibility against pyrethroids alone and was independent of existence of resistance to insecticides of other groups. This study emphasizes that appropriate rotation of different insecticides; including carbamates may prevent or delay the onset of resistance.