Larvicidal activity of few select indigenous plants of North East India against disease vector mosquitoes (Diptera: Culicidae)

Mosquitoes are the vectors of several life threatening diseases like dengue, malaria, Japanese encephalitis and lymphatic filariasis, which are widely present in the north-eastern states of India. Investigations on five local plants of north-east India, selected on the basis of their use by indigenous communities as fish poison, were carried out to study their mosquito larvicidal potential against Anopheles stephensi (malaria vector), Stegomyia aegypti (dengue vector) and Culex quinquefasciatus (lymphatic filariasis vector) mosquitoes. Crude Petroleum ether extracts of the roots of three plants viz. Derris elliptica, Linostoma decandrum and Croton tiglium were found to have remarkable larvicidal activity; D. elliptica extract was the most effective and with LC50 value of 0.307 μg/ml its activity was superior to propoxur, the standard synthetic larvicide. Half-life of larvicidal activity of D. elliptica and L. decandrum extracts ranged from 2-4 days.

Mosquito biodiversity of Dibru-Saikhowa biosphere reserve in Assam, India.

Entomological surveys were conducted for three consecutive years in core and buffer zone of the Dibru-Saikhowa biosphere reserve in pre monsoon, monsoon and post-monsoon seasons. A total of 52 species of mosquitoes under eleven genera have been detected. The genus Anopheles (18 species) was the predominant followed by Culex, Aedes, Mansonia, Armigeres, Mimomyia, Ochlerostatus, Malaya, Toxorhynchites, Ficalbia and Aedeomyia. The buffer zone of the forest reserve where human habitations are there exhibited the presence of maximum number of species (49 species under 10 genera) in comparison to core zone (42 species under 10 genera). In buffer zone, maximum numbers of species (38) were recorded in monsoon season followed by post- monsoon (35 species) and Pre-monsoon season (34 species). Whereas in core zone, maximum number of species were collected in post monsoon season followed by pre monsoon season and monsoon season. In Core and buffer zone, the maximum species were recorded from the ground pool habitat and slow flowing stream respectively. Among the disease vectors, the potential Japanese encephalitis vectors incriminated in India were very much prevalent. This study provides the list of available mosquito species recorded for the first time in the Dibru-Saikhowa biosphere reserve.

Community practices of using bed nets & acceptance & prospects of scaling up insecticide treated nets in north-east India.

BACKGROUND & OBJECTIVE: The National Vector Borne Disease Control Programme (NVBDCP), India has been promoting and scaling up the use of insecticide-treated nets (ITNs) in the tribal dominated malarious areas of north-east India. But, information on sleeping habits, bed net ownership and use practices, and feedback of communities in ITN--targeted areas is needed for formulating a strategic framework for upscaling the coverage of ITNs. We carried out a community-based cross-sectional survey in select areas of Nagaland and Mizoram (where ITNs were introduced) along with Assam (where ITNs were not introduced) to know the response of community. METHODS: Following large scale introduction of ITNs during 2001-2002 in the north-eastern States of Nagaland and Mizoram by NVBDCP, India, a cross-sectional community-based survey was undertaken in April-May, 2003 covering 435 households of Nagaland and 464 households in Mizoram, using a structured questionnaire, to assess the demographic variables relevant to bed net use, bed net washing practices and acceptability of ITNs etc., for upscaling the coverage of ITNs in the surveyed communities. A total of 448 households in 8 villages in a non-ITN PHC area of Assam were served as the control area. RESULTS: The average bet net usage per family (2.01 to 2.65). Bed net use was mostly seasonal in Nagaland where a majority (65.1-78.7%) used bed nets only during summers and monsoon as compared to the year round use in Mizoram (83.5%) and Assam (78.9%). Frequent washing of nets was most common in Assamese communities with 77 per cent households washing their nets at least once in a month. More than two third users favoured use of ITNs over the conventional indoor residual spray of DDT for malaria control. INTERPRETATION & CONCLUSION: ITNs are widely acceptable in the user communities and a demand for ITNs is evident in the non-user communities of north-east India. Approach of treating community owned nets through an efficient service delivery mechanism will be a viable option for upscaling the ITN coverage.

Community practices of using bed nets & acceptance & prospects of scaling up insecticide treated nets in north-east India.

Background & objectives: The National Vector Borne Disease Control Programme (NVBDCP), India has been promoting and scaling up the use of insecticide-treated nets (ITNs) in the tribal dominated malarious areas of north-east India. But, information on sleeping habits, bed net ownership and use practices, and feedback of communities in ITN - targeted areas is needed for formulating a strategic framework for upscaling the coverage of ITNs. We carried out a community-based cross-sectional survey in select areas of Nagaland and Mizoram (where ITNs were introduced) along with Assam (where ITNs were not introduced) to know the response of community. Methods: Following large scale introduction of ITNs during 2001-2002 in the north-eastern States of Nagaland and Mizoram by NVBDCP, India, a cross-sectional community-based survey was undertaken in April-May, 2003 covering 435 households of Nagaland and 464 households in Mizoram, using a structured questionnaire, to assess the demographic variables relevant to bed net use, bed net washing practices and acceptability of ITNs etc., for upscaling the coverage of ITNs in the surveyed communities. A total of 448 households in 8 villages in a non-ITN PHC area of Assam were served as the control area. Results: The average bet net usage per family (2.01 to 2.65). Bed net use was mostly seasonal in Nagaland where a majority (65.1-78.7%) used bed nets only during summers and monsoon as compared to the year round use in Mizoram (83.5%) and Assam (78.9%). Frequent washing of nets was most common in Assamese communities with 77 per cent households washing their nets at least once in a month. More than two third users favoured use of ITNs over the conventional indoor residual spray of DDT for malaria control. Interpretation & conclusion: ITNs are widely acceptable in the user communities and a demand for ITNs is evident in the non-user communities of north-east India. Approach of treating community owned nets through an efficient service delivery mechanism will be a viable option for upscaling the ITN coverage.

Detection & molecular confirmation of a focus of Plasmodium malariae in Arunachal Pradesh, India.

BACKGROUND & OBJECTIVE: During a malaria epidemiological study in Arunachal Pradesh, Plasmodium malariae like human malaria parasites were seen in blood smears from fever cases. The study was undertaken to detect the presence of P. malariae and to confirm its identity through DNA based polymerase chain reaction approach. METHODS: Fever survey was carried out in 22 villages in Indo-Myanmar bordering district of Lohit, Arunachal Pradesh in 2005. Morphologically suspected P. malariae cases were confirmed using nested PCR based on 18S small subunit ribosomal DNA gene sequence. RESULTS: Screening of 1,995 fever cases resulted in 9 probable cases of P. malariae based on morphological identification in Chakma tribe people residing in 2 villages. Nested PCR confirmed the identity of all probable cases of P. malariae by producing diagnostic band of 144 bp. PCR method was able to detect mixed infection of P. malariae with P. vivax and with P. falciparum. INTERPRETATION & CONCLUSION: P. malariae may have been present in Arunachal Pradesh but most probably is being misdiagnosed due to its close resemblance with P. vivax, especially in ring forms. Estimation of actual case load of P. malariae in north-east India is, therefore, important with accurate species identification using molecular methods.

Characterization of ITS2 rDNA of Anopheles philippinensis and Anopheles nivipes (Diptera: Culicidae) from north-east India.

Anopheles (Cellia) philippinensis Ludlow and Anopheles (Cellia) nivipes (Theobald) are two closely related, morphologically very similar, mosquito species in the Annularis group, which play a supportive role in malaria transmission in north-east India. We amplified and performed sequence analysis for the internal transcribed spacer 2 (ITS2) locus of ribosomal DNA (rDNA) gene of morphologically confirmed specimens of these two species from the states of Assam and Nagaland. An. philippinensis and An. nivipes shared 85.2% sequence similarity and no intra-species variation was found in the nucleotide sequences of the two species. Overall, sequence data of the ITS2 marker revealed that both these species from north-east India differed by as much as have been reported from specimens of eastern Thailand.

Potential of Anopheles philippinensis-nivipes complex mosquitoes as malaria vector in north-east India.

Anopheles philippinensis-nivipes complex mosquitoes, captured in outdoor human landing catches and light traps in human dwellings from four different sites in Assam state and adjoining areas, were examined (n=1670) for the presence of circumsporozoite antigen (CSA) through enzyme linked immunosorbent assays (ELISA), using species specific capture monoclonal antibodies, of Plasmodium falciparum and two Plasmodium vivax polymorphs (Pv 210 and VK 247). In ELISA, 28 pools were found positive for CSA that accounted for minimum sporozoite rate of 1.7% (95% CI 1.11-2.41). Twenty five percent (7/28) of the positive pools were reactive for P. falciparum and between the two polymorphs of P. vivax, VK 247 was predominant with 77% (20/26) of all P. vivax positive pools. Results were suggestive of most likely involvement of Anopheles philippinensis-nivipes complex mosquitoes in malaria transmission in north-east India.

Taxonomical observations on Anopheles philippinensis/nivipes group mosquitoes in north-east India.

To examine the distribution pattern of Anopheles philippinensis and An. nivipes, mosquitoes belonging to An. philippinensis/nivipes group were collected from five sites in north-east Indian state of Assam and its adjoining areas and identified. Presence of both An. philippinensis and An. nivipes, with predominance of the latter taxon, was recorded with conformity on the basis of pupal paddle characteristics. Based on adult wing characters, 91.4% specimens were identified as An. philippinensis and 8.6% as An. nivipes with gradually increasing proportion of An. nivipes in westwardly direction. An. philippinensis/ nivipes was noted to be mid night biter, mainly exophagic and exclusively exophilic. Of the two species, An. philippinensis appears to be mostly anthropophilic and available in broken forest areas whereas An. nivipes seems to be mostly zoophilic and generally found in ecotone zones in plains areas.

Malaria control in a forest camp in an oil exploration area of Upper Assam.

BACKGROUND: Assam, in north-east India, is extremely rich in hydrocarbon deposits and the oil industry is the major contributor to its economy. A large number of oil fields and related installations in Assam are located in forest areas or on their fringes where malaria is a serious problem among field staff and security personnel, adversely affecting oil production. We carried out an operational research study for one year in a forest-based industrial security camp of Dibrugarh district and developed an effective malaria control strategy for such areas. METHODS: The specific strategy was formulated and implemented after taking into account the local epidemiology of malaria, vector's ecology and malaria risk behaviour of the camp inmates. The strategy was based on reducing the man-vector contact, using deltamethrin-treated mosquito nets in conjunction with mosquito repellent cream and weekly chemoprophylaxis with 300 mg chloroquine. The impact of the strategy was monitored entomologically and epidemiologically for one year after implementation. RESULTS: The mean landing rate of Anopheles dirus, the vector mosquito in the camp area, was 5.03 per person per night during the monitoring. In spite of such a high density of the vector, the man-vector contact was effectively checked by the intervention measures adopted. As a result, the incidence of malaria in the camp was reduced by > 90% as compared to previous years and the number of malaria cases came down from 6.7 per 1000 man-nights in 1998-99 to 0.06 in 2000-01. Mortality due to malaria was completely eliminated. CONCLUSION: Control of malaria should be based on the local determinants of transmission. The use of insecticide-treated mosquito nets in conjunction with a mosquito repellent cream is a good intervention for controlling Anopheles dirus-transmitted malaria in the forests of north-east India. The control module developed on the principle of reducing man-mosquito contact is easy to implement, cost-effective and replicable in similar forest-based locations.

Physico-chemical characteristics of breeding habitats of Anopheles dirus (Diptera:Culicidae) in Assam, India.

Larval ecology of Anopheles dirus, the main vector of forest malaria in north-eastern region of India, was studied in relation to physico-chemical characteristics of its breeding habitats in a rain forest area of Assam, India. Shady stream side pools, positive for the breeding of An. dirus, had significantly higher amounts of total hardness (P < 0.024) in comparison to negative pools of similar type. When compared with An. dirus negative breeding habitats, in hot-wet as well as in cool-dry seasons, An. dirus positive shady ground pools showed higher mean values of total alkalinity, hardness and chloride content, whereas lower pH, dissolved oxygen along with higher total alkalinity and hardness were possessed by An. dirus positive stream side pools. Specificity of breeding habitats of An. dirus in relation to its ecology has been discussed.

Risk factors of malaria in the fringes of an evergreen monsoon forest of Arunachal Pradesh.

BACKGROUND: The forested hilly and foothill regions of north-east India are highly endemic for malaria and have a distinct epidemiological pattern. Nearly half the reported cases of malaria are from these areas. A knowledge of the risk factors in this eco-geographic entity may be helpful in formulating a specific control strategy. Hence, we conducted a community-based epidemiological study in a hilly, forested terrain of Arunachal Pradesh and examined different socio-demographic factors to identify those predisposing to the occurrence of malaria, especially Plasmodium falciparum infection, in such areas. METHODS: Four epidemiological surveys were carried out during 1997 in 7 villages located at the fringes of a forest (total population: 1177) under Nompong Primary Health Centre of Changlang district, Arunachal Pradesh. Blood slides were collected randomly from the inhabitants, irrespective of their fever status, ensuring at least 50% coverage. One hundred and thirty-four microscopically confirmed Plasmodium falciparum cases were identified and 536 controls were randomly selected from the list of uninfected inhabitants. Relevant socio-demographic information was obtained from both cases and controls. The data were analysed by simple and multiple logistic regression using the unconditional maximum likelihood method. RESULTS: Factors which were found to be strongly associated with Plasmodium falciparum malaria on univariate analysis were age, ethnicity, village of residence and accessibility to the nearest health care facility. However, in multiple regression analysis, after controlling for the effects of confounding variables, the only risk factor identified was accessibility to the nearest health care facility (adjusted odds ratio: 4.5; 95% CI: 1.8-11.3; p < 0.0001 for those at a distance of 2-8 km and adjusted odds ratio: 11.1; 95% CI: 4.1-30.0; p < 0.0001 for those > 8 km away). CONCLUSION: Distance, particularly non-motorable distance, from the place of residence to the nearest health care facility was a major risk factor for malaria in this hilly forested terrain. This indicates the need for special efforts to detect cases early and institute treatment promptly in such areas of the north-eastern region, so as to reduce the morbidity and mortality of malaria.

Epidemiology of malaria outbreak (April/May, 1999) in Titabor Primary Health Centre, district Jorhat (Assam).

An investigation was undertaken of a malaria outbreak in the Primary Health Centre Titabor, district Jorhat, Assam during May/June 1999. The fever rate in the community since March 1999, was 44.4 per cent with an average case load of 2.5 per family. The fever cases peaked in the third week of May. Slide positive and slide falciparum rates in mass blood survey, in the study village were 16.1 and 14.5 per cent respectively with 90 per cent infection of Plasmodium falciparum. Males (SPR 17.5%) suffered relatively more than females (SPR 14.7%). Malaria prevalence was significantly less in individuals above 15 yr of age (SPR 11.0%) as compared to those below 15 yr (SPR 22.9%). Prevalence of malaria as well as mosquito densities in different clusters of the village were inversely related to the distance from the forested Naga hills. Anopheles minimus and A. dirus were collected in good numbers with comparatively higher densities of the former. Several factors like unusual climatic conditions, inadequate surveillance, unsatisfactory laboratory services and inadequate indoor residual insecticide spray were instrumental for the outbreak.

Epidemiological importance of younger age group during malaria epidemic in PHC Tamulpur, Assam.

An investigation of a malaria epidemic was carried out in Tamulpur Primary Health Centre, district Nalbari, Assam during April 1995. The analysis revealed that children between 3 and 12 years of age who were treated and who recovered clinically from fever during the epidemic were instrumental in the progression of the epidemic by acting as Plasmodium falciparum gametocyte reservoirs. Special efforts are required for treatment of children below 12 years during an epidemic.

Breeding and day resting habitats of Anopheles dirus in Assam, India.

Longitudinal surveys at monthly frequency were carried out during 1995-1996 in a forest fringed village of district Dibrugarh, Assam to decipher the breeding and day resting habitats of Anopheles dirus. It regularly bred in small, shallow, rain filled, transient, shady or partly shady puddles/ground pools in the rainy months and in the perinneal streams in the adjoining forest of the village during dry months. In pools, the degree of interspecific association (0.238 +/- 0.174) and index of association (0.428) of An. dirus breeding was highest with Aedes caecus. An. dirus was completely exophilic and rested during day time in the forest mostly on tree trunks upto the height of 4-5 feet from the ground in dark moist niches avoiding direct sun light.

Feeding patterns of Anopheles dirus, the major vector of forest malaria in north east India.

The feeding behavior of Anopheles dirus, the forest breeding, major malaria vector in northeast India was studied. The analysis of blood meals collected from this mosquito revealed that the species was highly anthropophilic in nature, the anthropophilic index being 90.5. The results of bait collection on human and cattle bait also confirmed its biting preference for human hosts. The species was observed to land on human bait throughout the night, showing prominent biting time at 20.00-21.00, 23.00-24.00 and 02.00-03.00 hours during the study period.