Insecticide susceptibility of Phlebotomus argentipes sandflies, vectors of visceral leishmaniasis in India.

OBJECTIVES: Indoor residual spraying (IRS) with insecticides is the main vector control intervention for the elimination of visceral leishmaniasis in India. After a change in IRS policy in 2015 due to widespread resistance of Phlebotomus argentipes to DDT, IRS with DDT was replaced with alpha-cypermethrin IRS in 2016. The objective of the present study was to evaluate the susceptibility of P. argentipes to DDT and its alternatives, namely malathion and pirimiphos-methyl (organophosphates); alpha-cypermethrin, deltamethrin, lambda-cyhalothrin and permethrin (pyrethroids), and bendiocarb and propoxur (carbamates), in support of visceral leishmaniasis elimination in India. METHODS: Phlebotomus argentipes sandflies were collected from the visceral-leishmaniasis endemic states of Bihar, Jharkhand and West Bengal. In the WHO tube tests, the phenotypic susceptibility of F1, 2-day old, non-blood fed females were determined against filter papers impregnated with DDT 4%, malathion 5%, pirimiphos-methyl 0.25%, alpha-cypermethrin 0.05%, deltamethrin 0.05%, lambda-cyhalothrin 0.05%, permethrin 0.75%, bendiocarb 0.1% and propoxur 0.1%, which were sourced from Universiti Sains Malaysia. The knockdown of sandflies after 1-h exposure and mortality at 24 h after the 1-h exposure period were scored. RESULTS: Mean mortality of P. argentipes 24 h after exposure in tube tests was 22.6% for DDT and ≥ 98% for other insecticide-impregnated papers tested. CONCLUSION: Phlebotomus argentipes continues to be highly resistant to DDT with no reversal of resistance after DDT's withdrawal from IRS. P. argentipes was fully susceptible to pyrethroid, organophosphate and carbamate insecticides tested. Regular monitoring is warranted for insecticide resistance management in sandfly vectors.

Insecticide susceptibility of Phlebotomus argentipes & assessment of vector control in two districts of West Bengal, India.

BACKGROUND & OBJECTIVES: Kala-azar or visceral leishmanisis (VL) is known to be endemic in several States of India including West Bengal (WB). Only meager information is available on the vector dynamics of its vector species, Phlebotomus argentipes particularly in relation to control measure from this State. Hence, a pilot study was undertaken to assess the control strategy and its impact on vector in two endemic districts of WB, India. METHODS: Two villages each from the two districts, Maldah and Burdwan, were selected for the study. Seasonal variation of sandflies was observed during pre-monsoon, post-monsoon and winter seasons. Susceptibility test of P. argentipes against DDT and bioassay on DDT sprayed wall and on long lasting insecticide nets (LN) Perma Net [®] 2.0 were conducted as per the WHO standard methods. RESULTS: P. argentipes density was high during March to October. Susceptibility status of P. argentipes ranged from 40 to 61.54 per cent. Bioassay test showed 57.89 per cent mortality against LN PermaNet [®] -2.0. and 50 per cent against DDT on wall within 30 min of exposure. INTERPRETATION & CONCLUSIONS: Despite the integrated vector management approach, the sandfly population was high in the study area. The reason could be development of resistance in P. argentipes against DDT and low effectiveness of LN PermaNet [®] -2.0. The more pragmatic step will be to conduct large studies to monitor the susceptibility level in P. argentipes against DDT.

Spatial and temporal variation and hotspot detection of kala-azar disease in Vaishali district (Bihar), India.

BACKGROUND: An improved understanding in transmission variation of kala-azar is fundamental to conduct surveillance and implementing disease prevention strategies. This study investigated the spatio-temporal patterns and hotspot detection for reporting kala-azar cases in Vaishali district based on spatial statistical analysis. METHODS: Epidemiological data from the study area during 2007-2011 was used to examine the dynamic space-time pattern of kala-azar outbreaks, and all cases were geocoded at a village level. Spatial smoothing was applied to reduce random noise in the data. Inverse distance weighting (IDW) is used to interpolate and predict the pattern of VL cases distribution across the district. Moran's I Index (Moran's I) statistics was used to evaluate autocorrelation in kala-azar spatial distribution and test how villages were clustered or dispersed in space. Getis-Ord Gi*(d) was used to identify the hotspot and cold spot areas within the study site. RESULTS: Mapping kala-azar cases or incidences reflects the spatial heterogeneity in the incidence rate of kala-azar affected villages in Vaishali district. Kala-azar incidence rate map showed most of the highest endemic villages were located in southern, eastern and northwestern part of the district; in the middle part of the district generally show the medium occurrence of VL. There was a significant positive spatial autocorrelation of kala-azar incidences for five consecutive years, with Moran's I statistic ranging from 0.04-0.17 (P <0.01). The results revealed spatially clustered patterns with significant differences by village. The hotspots showed the spatial trend of kala-azar diffusion (P < 0.01). CONCLUSIONS: The results pointed to the usefulness of spatial statistical approach to improve our understanding the spatio-temporal dynamics and control of kala-azar. The study also showed the north-western and southern part of Vaishali district is most likely endemic cluster region. To employ exact and geographically suitable risk-reduction programmes, apply of such spatial analysis tools should suit a vital constituent in epidemiology research and risk evaluation of kala-azar.

User friendliness, efficiency & spray quality of stirrup pumps versus hand compression pumps for indoor residual spraying.

BACKGROUND & OBJECTIVES: Indoor residual spraying (IRS) is a proven tool to reduce visceral leishmaniasis vectors in endemic villages. In India IRS is being done with stirrup pumps, whereas Nepal, Bangladesh, and other countries use compression pumps. The present study was conducted with the objectives to compare the efficiency, cost and user friendliness of stirrup and compression pumps. METHODS: The study was carried out in Gorigawan village of the Vaishali district in north Bihar and included a total population of 3259 inhabitants in 605 households. Spraying with 50 per cent DDT was done by two teams with 6 persons per team under the supervision of investigators over 5 days with each type of pump (10 days in total using 2 stirrup pumps and 3 compression pumps) by the same sprayers in an alternate way. The spraying technique was observed using an observation check list, the number of houses and room surfaces sprayed was recorded and an interview with sprayers on their satisfaction with the two types of pumps was conducted. RESULTS: On average, 65 houses were covered per day with the compression pump and 56 houses were covered with the stirrup pump. The surface area sprayed per squad per day was higher for the compression pump (4636 m²) than for the stirrup pump (4102 m²). Observation showed that it was easy to maintain the spray swath with the compression pump but very difficult with the stirrup pump. The wastage of insecticide suspension was negligible for the compression pump but high for the stirrup pump. INTERPRETATION & CONCLUSIONS: The compression pump was found to be more user friendly due to its lower weight, easier to operate, lower operation cost, higher safety and better efficiency in terms of discharge rate and higher area coverage than the stirrup pump.

Oviposition behaviour of Phlebotomus argentipes--a laboratory-based study.

The breeding habitat of sandflies is a little studied and poorly understood phenomenon. More importantly, oviposition behaviour is a largely neglected aspect of sandfly biology and this knowledge gap further undermines our understanding of the biology of sandflies. Pheromones released by the eggs play an important role in identifying good sites for oviposition by female insects. Several recent studies have examined the oviposition pheromone. The present study provides a preliminary report on the oviposition behaviour of Phlebotomus argentipes, the only vector of kala-azar (or visceral leishmaniasis) on the Indian sub-continent. Sandflies prefer to oviposit their eggs on surfaces that contain organic substances, especially substances with an odour of decaying animal products and the remains of conspecific eggs. The results presented here suggest that the odour released by the organic substances of old sandfly colony remains that contain dead flies, old unhatched eggs, larval food containing vertebrate faeces, frass and other organic matter serves as an attractant for the ovipositing females of P. argentipes and hence greatly increases the number of oviposited eggs compared to eggs deposited in controlled oviposition pots. This result will be helpful in maintaining an efficient colony of P. argentipes and may be a promising tool for monitoring and controlling the target insect as part of a synergistic approach.

Appraisal of Phlebotomus argentipes habitat suitability using a remotely sensed index in the kala-azar endemic focus of Bihar, India.

Visceral leishmaniasis, or kala-azar, is recognised as a serious emerging public health problem in India. In this study, environmental parameters, such as land surface temperature (LST) and renormalised difference vegetation indices (RDVI), were used to delineate the association between environmental variables and Phlebotomus argentipes abundance in a representative endemic region of Bihar, India. The adult P. argentipes were collected between September 2009-February 2010 using the hand-held aspirator technique. The distribution of P. argentipes was analysed with the LST and RDVI of the peak and lean seasons. The association between environmental covariates and P. argentipes density was analysed a multivariate linear regression model. The sandfly density at its maximum in September, whereas the minimum density was recorded in January. The regression model indicated that the season, minimum LST, mean LST and mean RDVI were the best environmental covariates for the P. argentipes distribution. The final model indicated that nearly 74% of the variance of sandfly density could be explained by these environmental covariates. This approach might be useful for mapping and predicting the distribution of P. argentipes, which may help the health agencies that are involved in the kala-azar control programme focus on high-risk areas.

Delimitation of kala-azar risk areas in the district of Vaishali in Bihar (India) using a geo-environmental approach.

Remote sensing and geographical information technologies were used to discriminate areas of high and low risk for contracting kala-azar or visceral leishmaniasis. Satellite data were digitally processed to generate maps of land cover and spectral indices, such as the normalised difference vegetation index and wetness index. To map estimated vector abundance and indoor climate data, local polynomial interpolations were used based on the weightage values. Attribute layers were prepared based on illiteracy and the unemployed proportion of the population and associated with village boundaries. Pearson's correlation coefficient was used to estimate the relationship between environmental variables and disease incidence across the study area. The cell values for each input raster in the analysis were assigned values from the evaluation scale. Simple weighting/ratings based on the degree of favourable conditions for kala-azar transmission were used for all the variables, leading to geo-environmental risk model. Variables such as, land use/land cover, vegetation conditions, surface dampness, the indoor climate, illiteracy rates and the size of the unemployed population were considered for inclusion in the geo-environmental kala-azar risk model. The risk model was stratified into areas of "risk"and "non-risk"for the disease, based on calculation of risk indices. The described approach constitutes a promising tool for microlevel kala-azar surveillance and aids in directing control efforts.

Impact of intensified control on visceral leishmaniasis in a highly-endemic district of Bihar, India: an interrupted time series analysis.

Visceral leishmaniasis (VL) is declining in India and the World Health Organization's (WHO) 2020 'elimination as a public health problem' target has nearly been achieved. Intensified combined interventions might help reach elimination, but their impact has not been assessed. WHO's Neglected Tropical Diseases 2021-2030 roadmap provides an opportunity to revisit VL control strategies. We estimated the combined effect of a district-wide pilot of intensified interventions in the highly-endemic Vaishali district, where cases fell from 3,598 in 2012-2014 to 762 in 2015-2017. The intensified control approach comprised indoor residual spraying with improved supervision; VL-specific training for accredited social health activists to reduce onset-to-diagnosis time; and increased Information Education & Communication activities in the community. We compared the rate of incidence decrease in Vaishali to other districts in Bihar state via an interrupted time series analysis with a spatiotemporal model informed by previous VL epidemiological estimates. Changes in Vaishali's rank among Bihar's endemic districts in terms of monthly incidence showed a change pre-pilot (3rd highest out of 33 reporting districts) vs. during the pilot (9th) (p<1e-10). The rate of decline in Vaishali's incidence saw no change in rank at 11th highest, both pre-pilot & during the pilot. Counterfactual model simulations suggest an estimated median of 352 cases (IQR 234-477) were averted by the Vaishali pilot between January 2015 and December 2017, which was robust to modest changes in the onset-to-diagnosis distribution. Strengthening control strategies may have precipitated a substantial change in VL incidence in Vaishali and suggests this approach should be piloted in other highly-endemic districts.

A comparative evaluation of end-emic and non-endemic region of visceral leishmaniasis (Kala-azar) in India with ground survey and space technology.

In visceral leishmaniasis, phlebotomine vectors are targets for control measures. Understanding the ecosystem of the vectors is a prerequisite for creating these control measures. This study endeavours to delineate the suitable locations of Phlebotomus argentipes with relation to environmental characteristics between endemic and non-endemic districts in India. A cross-sectional survey was conducted on 25 villages in each district. Environmental data were obtained through remote sensing images and vector density was measured using a CDC light trap. Simple linear regression analysis was used to measure the association between climatic parameters and vector density. Using factor analysis, the relationship between land cover classes and P. argentipes density among the villages in both districts was investigated. The results of the regression analysis indicated that indoor temperature and relative humidity are the best predictors for P. argentipes distribution. Factor analysis confirmed breeding preferences for P. argentipes by landscape element. Minimum Normalised Difference Vegetation Index, marshy land and orchard/settlement produced high loading in an endemic region, whereas water bodies and dense forest were preferred in non-endemic sites. Soil properties between the two districts were studied and indicated that soil pH and moisture content is higher in endemic sites compared to non-endemic sites. The present study should be utilised to make critical decisions for vector surveillance and controlling Kala-azar disease vectors.

Kala-azar elimination in a highly-endemic district of Bihar, India: A success story.

BACKGROUND: Visceral leishmaniasis (VL) or Kala-azar has been a major public health problem in Bihar, India, for several decades. A few VL infected districts including Vaishali have reported >600 cases annually. Hence, in 2015, the Government of India entrusted ICMR-Rajendra Memorial Research Institute of Medical Sciences, Patna, to implement an integrated control strategy for achieving the VL elimination target (<1 case per 10,000 people at the block level) in the Vaishali District of Bihar. METHODOLOGY: This study was conducted between January 2015 and December 2016. An integrated control strategy including the spatio-temporal mapping of VL-case distribution, active case detection, chemical-based vector control using indoor residual spraying (IRS), community awareness campaigns, the training of IRS members, the training of medical doctors for effective treatment, daily monitoring and the supervision of IRS activities, logistic management, post-IRS quality assurance, epidemiological surveillance, and entomological monitoring was performed. An insecticide quantification test was performed for evaluating the IRS quality on sprayed walls. A modern compression pump was used to maintain spray quality on different wall surfaces. The impact of IRS was assessed through sand fly collection in human dwellings and cattle sheds in pre- and post-IRS. The insecticide susceptibility of local P. argentipes was performed before each IRS round (in February and June) during 2015-2016. Statistical analysis such as the mean, percentage, and 95% CI were used to summarize the results. FINDINGS: All 16 blocks of the Vaishali District achieved the VL elimination target in 2016. The integrated VL control strategy helped reduce the number of VL cases from 664 in 2014 to 163 in 2016 and the number of endemic villages from 282 in 2014 to 142 in 2016. The case reduction rate was increased from 22.6% in 2014 to 58.8% in 2016. On average, 74 VL infected villages became Kala-azar free each year from 2015 to 2016. CONCLUSIONS: The results of this study suggest that the elimination of VL is possible from all endemic blocks of Bihar if the integrated Vaishali VL control strategy is applied under strong monitoring and supervision.

Assessing the combined effects of household type and insecticide effectiveness for kala-azar vector control using indoor residual spraying: a case study from North Bihar, India.

BACKGROUND: Indoor residual spraying (IRS) is the mainstay for vector control intervention of visceral leishmaniasis (VL) in India. Little is known on the control effects of IRS on different household types. Here, we assessed if IRS with insecticides has an equal residual and interventional effect on all household types in a village. We also developed a combined spatial-risk map and a sand fly, Phlebotomus argentipes density analytical model based on household characteristics, insecticide susceptibility and IRS-status to explore the spatio-temporal distributions of the vector at a micro-scale level. METHODS: This study was carried out in two villages of Mahnar block in Vaishali district, Bihar. IRS using two insecticides [dichlorodiphenyltrichloroethane (DDT 50%) and synthetic pyrethroid (SP 5%)] was evaluated for VL-vector (P. argentipes) control. Temporal residual efficacy of the insecticides on different wall-surface types was evaluated using the cone-bioassay technique according to WHO guidelines. Insecticide susceptibility of local P. argentipes was explored using the tube-bioassay method. Pre- and post-IRS sand fly densities were monitored in human dwellings and animal shelters using Centers for Disease Control light-traps installed between 18:00-6:00 h. A best-fit model for sand fly density analysis was developed using multiple logistic regression analysis. Geographical information system based spatial analysis techniques were employed to map the household type distribution of insecticide susceptibility of the vector, and IRS-status of the households to interpret the spatio-temporal distributions of P. argentipes. RESULTS: Phlebotomus argentipes was highly susceptible to SP (100%) but showed high resistance to DDT with a 49.1% mortality rate. SP-IRS has been reported as having better community acceptance than DDT-IRS in all household types. Residual efficacies were varied between wall-surfaces; both insecticides failed to achieve the duration of IRS effectiveness recommended by the WHO. Reduction in P. argentipes counts due to SP-IRS was higher than DDT-IRS between household groups (i.e. sprayed and sentinel), in all intervals post-IRS. Combined spatial risk-maps revealed a better control effect of SP-IRS on sand flies than DDT-IRS in all household types risk-zones. The multilevel logistic regression analysis explored five risk-factors that were strongly associated with the density of P. argentipes. CONCLUSIONS: The results contribute to furthering current understanding of IRS-practices for control of visceral leishmaniasis in endemic Bihar, which may help in future actions for improvements.

Trends in spatio-temporal dynamics of visceral leishmaniasis cases in a highly-endemic focus of Bihar, India: an investigation based on GIS tools.

BACKGROUND: Visceral leishmaniasis (VL) in Bihar State (India) continues to be endemic, despite the existence of effective treatment and a vector control program to control disease morbidity. A clear understanding of spatio-temporal distribution of VL may improve surveillance and control implementation. This study explored the trends in spatio-temporal dynamics of VL endemicity at a meso-scale level in Vaishali District, based on geographical information systems (GIS) tools and spatial statistical analysis. METHODS: A GIS database was used to integrate the VL case data from the study area between 2009 and 2014. All cases were spatially linked at a meso-scale level. Geospatial techniques, such as GIS-layer overlaying and mapping, were employed to visualize and detect the spatio-temporal patterns of a VL endemic outbreak across the district. The spatial statistic Moran's I Index (Moran's I) was used to simultaneously evaluate spatial-correlation between endemic villages and the spatial distribution patterns based on both the village location and the case incidence rate (CIR). Descriptive statistics such as mean, standard error, confidence intervals and percentages were used to summarize the VL case data. RESULTS: There were 624 endemic villages with 2719 (average 906 cases/year) VL cases during 2012-2014. The Moran's I revealed a cluster pattern (P < 0.05) of CIR distribution at the meso-scale level. On average, 68 villages were newly-endemic each year. Of which 93.1% of villages' endemicity were found to have occurred on the peripheries of the previous year endemic villages. The mean CIR of the endemic villages that were peripheral to the following year newly-endemic villages, compared to all endemic villages of the same year, was higher (P < 0.05). CONCLUSION: The results show that the VL endemicity of new villages tends to occur on the periphery of villages endemic in the previous year. High-CIR plays a major role in the spatial dispersion of the VL cases between non-endemic and endemic villages. This information can help achieve VL elimination throughout the Indian subcontinent by improving vector control design and implementation in highly-endemic district.

Spatial Distribution of Phlebotomus argentipes (Diptera: Psychodidae) in Eastern India, a Case Study Evaluating Multispatial Resolution Remotely Sensed Environmental Evidence and Microclimatic Data.

Remote sensing, a powerful tool for analyzing landscape factors, is being used to explore the spatial ecology of vectors of several diseases. This study aims to explore the role of buffer size in identification and quantification of geo-environmental factors from multispatial resolution satellite data and its application along with microclimatic data to kala-azar vector abundance modeling.Sand fly abundance and microclimatic data were collected from 210 sample sites during the premonsoon and postmonsoon season of 2014 from Muzaffarpur district of Bihar (India). Linear imaging self-scanning sensor (LISS-III; 23.5 m) and advanced wide field sensor (AWiFS; 56 m) imageries were used for generating environmental variables at 300- and 500-m buffer zones. Four analytical models of sand fly density were developed and evaluated for predictive accuracy.A total of 33 geo-environmental and four microclimatic variables were tested for the prediction of sand fly density, of which the best four were maximum temperature, relative humidity, Euclidean nearest-neighbor distance of settlement area to mixed bush-grass land, and surface water body. Predictive accuracy of the LISS-III models was found to be higher than AWiFS models at all buffer sizes.The results show that geo-environmental parameters and microclimatic data are the best predictors for sand fly density modeling. Buffer sizes play an important role in identifying the explanatory variables. Model parameters may be useful in identifying predisposing factors of sand fly habitat suitability at the micro level.

Studying DDT Susceptibility at Discriminating Time Intervals Focusing on Maximum Limit of Exposure Time Survived by DDT Resistant Phlebotomus argentipes (Diptera: Psychodidae): an Investigative Report.

Extensive application of routine insecticide i.e., dichlorodiphenyltrichloroethane (DDT) to control Phlebotomus argentipes (Diptera: Psychodidae), the proven vector of visceral leishmaniasis in India, had evoked the problem of resistance/tolerance against DDT, eventually nullifying the DDT dependent strategies to control this vector. Because tolerating an hour-long exposure to DDT is not challenging enough for the resistant P. argentipes, estimating susceptibility by exposing sand flies to insecticide for just an hour becomes a trivial and futile task.Therefore, this bioassay study was carried out to investigate the maximum limit of exposure time to which DDT resistant P. argentipes can endure the effect of DDT for their survival. The mortality rate of laboratory-reared DDT resistant strain P. argentipes exposed to DDT was studied at discriminating time intervals of 60 min and it was concluded that highly resistant sand flies could withstand up to 420 min of exposure to this insecticide. Additionally, the lethal time for female P. argentipes was observed to be higher than for males suggesting that they are highly resistant to DDT's toxicity. Our results support the monitoring of tolerance limit with respect to time and hence points towards an urgent need to change the World Health Organization's protocol for susceptibility identification in resistant P. argentipes.

Spatial distribution of P. argentipes in association with agricultural surrounding environment in North Bihar, India.

INTRODUCTION: Phlebotomus argentipes is considered to be one of the major control measure targets in eradicating visceral leishmaniasis (VL). This study demonstrates the spatial association of P. argentipes abundance in relation to agricultural environment in the endemic area of north Bihar. METHODOLOGY: Vector data were collected from the 95 villages of the study area using a handheld aspiration technique. Space technology and ground observationwere made to estimate the environmental characteristics. RESULTS: A total of 1,663 P. argentipes were collected between April and November 2011. Results showed negative and significant association between P. argentipes density and proximity to the agricultural land in both the dry (r = -0.39) and wet seasons (r = -0.55), respectively. A strong and positive association was observed between relative humidity and P. argentipes abundance (r = 0.55). Results illustrated a very strong positive association between soil moisture and P. argentipes abundance (r = 0.58). The agricultural land class density and the spatial abundance of P. argentipes showed a very strong and positive association (r = 0.46). CONCLUSIONS: This study will help in understanding the advantage of agricultural land use practices to delineate of P. argentipes habitat suitability, which may strengthen existing control strategies in this endemic area.

Seasonal relationship between normalized difference vegetation index and abundance of the Phlebotomus kala-azar vector in an endemic focus in Bihar, India.

Remote sensing was applied for the collection of spatio-temporal data to increase our understanding of the potential distribution of the kala-azar vector Phlebotomus argentipes in endemic areas of the Vaishali district of Bihar, India. We produced monthly distribution maps of the normalized difference vegetation index (NDVI) based on data from the thematic mapper (TM) sensor onboard the Landsat-5 satellite. Minimum, maximum and mean NDVI values were computed for each month and compared with the concurrent incidence of kala-azar and the vector density. Maximum and mean NDVI values (R2 = 0.55 and R2 = 0.60, respectively), as well as the season likelihood ratio (X2 = 17.51; P <0.001), were found to be strongly associated with kala-azar, while the correlation with between minimum NDVI values and kala-azar was weak (R2 = 0.25). Additionally, a strong association was found between the mean and maximum NDVI values with seasonal vector abundance (R2 = 0.60 and R2 = 0.55, respectively) but there was only a marginal association between minimum NDVI value and the spatial distribution of kala-azar vis-à-vis P. argentipes density.

Telehealth: a perspective approach for visceral leishmaniasis (kala-azar) control in India.

Visceral leishmaniasis, also known as kala-azar, is a vector borne disease caused by the protozoan parasite, L. donovani. Poor and neglected populations in Indian sub-continent are particularly affected by this disease. Due to the diversity of epidemiological situations, no single diagnosis, treatment, or control will be suitable for all. Control measures through case findings, treatment, and vector control are seldom used, even where they could be useful. Modern tools like telehealth, using space technology, have now come in handy to address issues of disease surveillance, control checking, and evaluation. The present study focuses on telehealth as a current vector control strategy, perspectives on diagnosis, treatment, and control of visceral leishmaniasis as these deserve more attention and research.

Localization of kala-azar in the endemic region of Bihar, India based on land use/land cover assessment at different scales.

Land cover, a critical variable in the epidemiology of kala-azar, can be remotely characterized by customizing and integrating "state-of-the-art" imagery at different spatial scales from different sensors onboard satellites. A study was conducted at four levels (national, state, district and village) investigating the role of land use/land cover (LULC) for leishmaniasis transmission resulting in a framework highlighting the links between LULC and areas endemic for the disease. Distribution maps were analysed by a probabilistic approach (Bayesian classifier) which produced a set of "suitability estimates" based on the probability of sand fly presence. The development of a sound knowledge of each link in the predicted sequence of satellite views offering an extraordinary opportunity to support the mapping of kala-azar endemicity and stratification of areas suitable for sand fly habitats across the country as well as at the local scale.

Role of remote sensing, geographical information system (GIS) and bioinformatics in kala-azar epidemiology.

Visceral leishmaniasis or kala-azar is a potent parasitic infection causing death of thousands of people each year. Medicinal compounds currently available for the treatment of kala-azar have serious side effects and decreased efficacy owing to the emergence of resistant strains. The type of immune reaction is also to be considered in patients infected with Leishmania donovani (L. donovani). For complete eradication of this disease, a high level modern research is currently being applied both at the molecular level as well as at the field level. The computational approaches like remote sensing, geographical information system (GIS) and bioinformatics are the key resources for the detection and distribution of vectors, patterns, ecological and environmental factors and genomic and proteomic analysis. Novel approaches like GIS and bioinformatics have been more appropriately utilized in determining the cause of visearal leishmaniasis and in designing strategies for preventing the disease from spreading from one region to another.

Incidence of visceral leishmaniasis in the Vaishali district of Bihar, India: spatial patterns and role of inland water bodies.

The role of the distribution of inland water bodies with respect to the transmission of visceral leishmaniasis (VL) and its dominant vector, Phlebotomous argentipes, has been studied at the regional scale in Bihar, eastern India. The Landsat TM sensor multispectral scanning radiometer, with a spatial resolution of 30 m in the visible, reflective-infrared and shortwave-infrared (SWIR) bands, was used to identify water bodies using the normalized differential pond index (NDPI) calculated as follows: (Green - SWIR I)/(Green + SWIR I). Nearest neighbour and grid square statistics were used to delineate spatial patterns and distribution of the sandfly vector and the disease it transmits. The female P. argentipes sandfly was found to be associated with the distance from open water and particularly abundant near non-perennial river banks (68.4%; P <0.001), while its association with rivers was focused further away from the water source (χ(2) = 26.3; P <0.001). The results also reveal that the distribution of VL is clustered around non-perennial riverbanks, while the pattern is slightly random around the perennial river banks. The grid square technique illustrate that the spatial distribution of the disease has a much stronger correlation with lower density of open waters surfaces as well as with sandfly densities (χ(2) = 26.0; P <0.001). The results of our study suggest that inland water presence poses a risk for VL by offering suitable breeding sites for P. argentipes, a fact that should be taken into account when attempting to control disease transmission.