Insecticide susceptibility status of malaria vectors, Anopheles culicifacies, Anopheles fluviatilis and Anopheles minimus in the tribal districts of Jharkhand state of India.

BACKGROUND AND OBJECTIVES: Insecticide resistance in malaria vectors has been a major challenge to vector control programs and updated information to the commonly used insecticides is essential for planning appropriate vector control measures. Anopheles culicifacies and An. fluviatilis are the two main vectors prevalent in Jharkhand state of India and role of An. minimus is contemplated in the transmission of malaria in this state. All the districts in the state are predominantly inhabited by the tribal population and are endemic for malaria. A study was undertaken in 12 districts of Jharkhand state to determine the insecticide susceptibility status of the 3 prevalent primary vector species, An. culicifacies, An. fluviatilis, and An. minimus. METHODS: Wild-caught adult female An. culicifacies, An. fluviatilis, and An. minimus, mosquitoes were collected from stratified ecotypes from different localities of 12 tribal districts of Jharkhand state during 2018 and 2019. Susceptibility tube tests were conducted following the WHO method using test kits. Mosquitoes were exposed to WHO impregnated papers with the prescribed discriminatory dosages of DDT - 4%, malathion - 5%, deltamethrin - 0.05%, permethrin - 0.75%, cyfluthrin - 0.15% and lambda cyhalothrin - 0.05%. RESULTS: Results indicated that An. culicifacies has developed multiple insecticide resistance in all the 12 districts of Jharkhand state. An. fluviatilis was reported resistant for the first time to DDT in all the districts but was susceptible to malathion, deltamethrin, and permethrin whereas in one district it showed possible resistance to malathion. An. minimus was studied in Noamundi CHC of West Singhbhum district, showed possible resistance against DDT but was susceptible to malathion, deltamethrin, and permethrin. INTERPRETATION & CONCLUSION: The development of multiple insecticide resistance in An. culicifacies including to pyrethroids, has been a concern for malaria control programmes for effective vector management but a report of resistance to DDT for the first time in An. fluviatlis in all the districts in the state is alarming, An. minimus was found in possible resistance category to DDT in one district and both the species were reported susceptible to malathion, deltamethrin, and permethrin. The result of the present study indicates a need for regular monitoring to assess the insecticide susceptibility to formulate effective vector control measures and resistance management.

Prevalence of sibling-species of Anopheles (Cellia) fluviatilis complex in Himachal Pradesh, India.

BACKGROUND & OBJECTIVES: Malaria is one of the most infectious and life-threatening vector borne disease in the tropics. Climate change can significantly influence malaria epidemiology and expansion of malaria vectors to hilly regions of Himachal Pradesh in India, hitherto considered areas of low transmission. Entomological surveillance in Kangra district of Himachal Pradesh revealed high density of a proven efficient vector of malaria, Anopheles fluviatilis, but transmission intensity of malaria was found very low. It was therefore considered prudent to investigate the sibling-species composition of An. fluviatilis complex in Kangra valley to ascertain their role in transmission of malaria. METHODS: The study was undertaken in six villages in Kangra district of Himachal Pradesh, India. A total of 4446 mosquitoes were collected during the one-year study period (2018) and processed in pools of ten for molecular characterization. DNA extraction and multiplex PCR was performed on 900 An. fluviatilis mosquitoes for differentiation of sibling-species. ELISA was used to detect Plasmodium falciparum and Plasmodium vivax circumsporozoite proteins in 3790 An. fluviatilis samples. RESULTS: Among prevalent mosquito species, An. fluviatilis was the predominant species constituting 69.5% of total mosquito collection. Sibling-species U was found in 92.22% and species T in 7.78% samples assayed. ELISA confirmed the absence of evidence of malaria parasite in any of the An. fluviatilis mosquitoes screened. Based on the difference in the sequences of conserved regions of the 28SrDNA, sibling-species U was confirmed as prevalent in the study villages. INTERPRETATION & CONCLUSION: Study revealed that in Kangra district, An. fluviatilis sibling-species U is predominant followed by species T, and both are non-vectors. The absence of malaria parasite and zoophagic nature of An. fluviatilis established through blood meal analysis, confirmed that both U and T are non-vector sibling-species.

Larvicidal potential of essential oils against Musca domestica and Anopheles stephensi.

The larvicidal activity of Mentha piperita, Cymbopogan citratus (lemongrass), Eucalyptus globulus and Citrus sinensis (orange) essential oils and their combinations was evaluated against Musca domestica (housefly) and Anopheles stephensi (mosquitoes) through contact toxicity assay. Among all the tested essential oils/combinations, Me. piperita was found to be the most effective larvicidal agent against Mu. domestica and An. stephensi with LC50 values of 0.66 µl/cm(2) and 44.66 ppm, respectively, after 48 h. The results clearly highlighted that the addition of mentha oil to other oils (1:1 ratio) improved their larvicidal activity. The order of effectiveness of essential oils/combinations indicated that the pattern for An. stephensi follows the trend as mentha > mentha + lemongrass > lemongrass > mentha + eucalyptus > eucalyptus > mentha + orange > orange and for Mu. domestica as mentha > mentha + lemongrass > lemongrass > mentha + orange > orange > mentha + eucalyptus > eucalyptus. The images obtained from scanning electron microscopy (SEM) analysis indicated the toxic effect of Me. piperita as the treated larvae were observed to be dehydrated and deformed. This study demonstrates the effectiveness of tested essential oils/combinations against the larval stages of Mu. domestica and An. stephensi and has the potential for development of botanical formulations.

Impact of container material on the development of Aedes aegypti larvae at different temperatures.

BACKGROUND & OBJECTIVES: Aedes aegypti, the primary vector of dengue generally breeds in intradomestic and peridomestic containers made up of different materials, i.e. plastic, iron, rubber, earthen material etc. The material of container is likely to affect the temperature of water in container with variation in environmental temperature. The present study was aimed to determine the effect of different container materials on larval development of Ae. aegypti at different temperatures. METHODS: Newly hatched I instar larvae (2-4 h old) were used in the study and experiments were conducted using three different containers made up of plastic, iron and earthen material. Three replicates for each type of container at 22, 26, 30, 34, 38, 40, and 42°C were placed in environmental chamber for the development of larvae. RESULTS: At temperatures >22°C, 50% pupation was completed in earthen pot within 4.3±0.6 to 6.3±0.6 days followed by plastic containers (5±0 to 8±0 days) and iron containers (6±0 to 9±0 days). Developmental time for 50% pupation in the three containers differed significantly (p < 0.05) at all the experimental temperature ranges. A significant variation was found in the temperature of environmental chamber and the temperature of water in three types of containers (p < 0.05). The difference in the temperatures of water in different containers resulted in significant variations in the developmental period of larvae. More than 35°C temperature of water was found inimical for pupal development. INTERPRETATION & CONCLUSION: The results revealed the variation in temperature of water in different types of containers depending on the material of container, affecting duration of larval development. As the larval development was faster in earthen pot as compared to plastic and iron containers, community should be discouraged for storing the water in earthen pots. However, in view of containers of different materials used by the community in different temperature zones in the country, further studies are required for devising area-specific preventive measures for Aedes breeding.

Anopheline mosquitoes in District Ramgarh (Jharkhand), India.

BACKGROUND & OBJECTIVES: Jharkhand is one of the highly malaria endemic states in India and experiencing vast ecological and human-induced changes over the years. These changes have provided more favourable conditions for malaria transmission in the region. The present study was carried out to find out the distribution and prevalence of anopheline vector and non-vector species in District Ramgarh of Jharkhand state. METHODS: Daytime indoor resting adult female anopheline mosquitoes were collected from four subcentres comprising of eight study villages in District Ramgarh. The collections were made from fixed as well as random human dwellings and cattlesheds on fortnightly basis using manual aspiration method from January to December 2012. Mosquito identification was done by using standard identification keys. RESULTS: A total of 18,875 anophelines belonging to 19 species were collected. Of these, 61.87% were vector species (An. culicifacies, An. fluviatilis and An. annularis). Of total vector collection, 57.44% was observed in Gola block and 42.55% in Ramgarh. An. culicifacies was predominant species followed by An. fluviatilis and An. annularis in the study area. Out of 19, eight anopheline species exhibited successional changes in their composition over the period of years. Statistical analysis revealed positive correlation between meteorological variables and man hour density in case of An. culicifacies, whereas these were negatively correlated in case of An. fluviatilis and An. annularis. INTERPRETATION & CONCLUSION: The study revealed the prevalence of three recognised malaria vector species (An. culicifacies, An. fluviatilis and An. annularis) in high density throughout the year in this area, which indicates possibility of widening of malaria transmission window in the presence of malaria parasites. The shifting of anopheline species in Ramgarh also indicate alteration in ecological, environmental and sociological conditions, which necessitate routine monitoring on ecology and successional changes of vector species as well as malariological survey for management and adoption of appropriate vector control strategies in this area.

Bionomics and vectorial capacity of Anopheles annularis with special reference to India: a review.

Anopheles annularis is widely distributed mosquito species all over the country. An. annularis has been incriminated as a malaria vector in India, Sri Lanka, Bangladesh, Myanmar, Indonesia, Malaysia and China. In India, it has been reported to play an important role in malaria transmission as a secondary vector in certain parts of Assam, West Bengal and U.P. In Odisha and some neighbouring countries such as Sri Lanka, Nepal and Myanmar it has been recognised as a primary vector of malaria. This is a species complex of two sibling species A and B but the role of these sibling species in malaria transmission is not clearly known. An. annularis is resistant to DDT and dieldrin/HCH and susceptible to malathion and synthetic pyrethorides in most of the parts of India. In view of rapid change in ecological conditions, further studies are required on the bionomics of An. annularis and its role in malaria transmission in other parts of the country. Considering the importance of An. annularis as a malaria vector, the bionomics and its role in malaria transmission has been reviewed in this paper. In this communication, an attempt has been made to review its bionomics and its role as malaria vector. An. annularis is a competent vector of malaria, thus, due attention should be paid for its control under the vector control programmes specially in border states where it is playing a primary role in malaria transmission.

Insecticide susceptibility status of Phlebotomus argentipes, a vector of visceral leishmaniasis in different foci in three states of India.

BACKGROUND & OBJECTIVES: Phlebotomus argentipes is the vector for visceral leishmaniasis in India. The development of resistance in kala-azar vector to DDT has been reported from various parts of India. The main objective of this study was to generate information on insecticides susceptibility status of P. argentipes to DDT, malathion and deltamethrin in different parts in three states of India. METHODS: Phlebotomus argentipes were collected from different villages, identified and used to investigate the susceptibility status against DDT, malathion and deltamethrin as per the WHO standard methods. RESULTS: Phlebotomus argentipes was resistant to DDT in different areas, viz. PHCs Murumgaon in Maharashtra; Ramgarh in Jharkhand; Kodah, Falka, Mahua and Lalganj in Bihar. In Phulwari Shareef PHC of Patna district in Bihar, DDT produced 89% mortality in P. argentipes, indicating resistant/tolerance (verification required) to DDT. The corrected percent mortality to malathion (5%) in different areas ranged between 98 and 100%; and to deltamethrin (0.05%) between 98.4 and 100%. The results showed that the tested P. argentipes are susceptible to malathion and deltamethrin. CONCLUSION: Phlebotomus argentipes are still susceptible to malathion and deltamethrin, but resistant to DDT. The susceptibility status of P. argentipes should be monitored regularly in diversified situations to ascertain the judicious use of insecticides being used for indoor residual spraying in the programme for rational use of appropriate insecticide.

Evaluation of indoor residual spray and insecticide treated bed nets in a malaria endemic area of Santhal Pargana, Dumka district (Jharkhand).

The study was carried out for evaluation of various activities of malaria control programme in five different tribal and malaria endemic Primary Health Centres of Dumka district (Jharkhand) during 2007-08. A total of 321 houses of 18 villages were surveyed on use of indoor residual spray (IRS) and insecticide-treated bed nets (ITN) and other activities as tool for vector control and interrupting the transmission of malaria. Out of 690 living rooms and 343 verandahs examined, IRS with Dichlorodiphenyl-trichloro-ethane (DDT) was done only in 16.23% living rooms and 64.72% verandahs. Refusal rate of IRS in living rooms was 81.93% due to lack of knowledge regarding the importance of IRS, no prior information to villagers, houses locked, reluctance to remove domestic articles, dislike of smell of DDT spray. Compliance rate of ITN uses was 71.66% during the night, which might be a factor for decline in malaria cases in the study area. Various important components of the programme, viz. surveillance and compliance to treatment activities, use of rapid diagnostic test kits (RDKs), involvement of accredited social health activist (ASHA's) and fever treatment depots (FTDs), laboratory activities, adult mosquito collection, other activities like constitution of village health sanitation committee, information education and communication activities, capacity building, use of larvivorous fishes, supervision of IRS etc. require much strengthening. However, 100% community acceptance was recorded for ITN in the villages surveyed. In addition, an entomological study was carried out for information on prevalence of mosquito species in this area to find out effectiveness of IRS activities. Eleven anopheline species, including three malaria vectors i.e., An. culicifacies, An. stephensi, An. fluviatilis, An. annularis, An. subpictus, An. nigerrimus, An. pallidus, An. aconitus, An. vagus, An. jamsii and An. splendidus were collected from cattle and human dwellings.

Prevalence of Aedes aegypti Linnaeus and Aedes albopictus Skuse in Koderma, Jharkhand.

Entomological survey was carried out in different localities of Koderma district of Jharkhand with a view to study the prevalence, distribution and stratification of areas for Aedes mosquito species. A total of 233 houses were covered during house to house larval and adult survey. Aedes breeding could be detected in 157 houses. In all, a total of 942 domestic water containers were searched, out of which 461 were found positive. The overall house index(HI) container index(CI) breteau index(B1) and pupal index(PI) were 67.38%, 48.94%, 197.85% and 79.4%, respectively. The survey revealed that Aedes aegypti Linnaeus and Aedes albopictus Skuse are well established in Koderma with most of the areas showing high adult and larval indices. The preventive strategy needs to be directed towards minimizing the breeding potential of Aedes and water management practice by individuals along with implementation of urban bye-laws as well as IEC activities to contain Aedes breeding in future.

Situation analysis of malaria in Godda district of Jharkhand during malaria epidemic.

The epidemiological and entomological investigations were carried out in 18 tribal villages of Godda district of Jharkhand state revealed average slide positivity rate (SPR) 9.9% and slide positivity rate for Plasmodium falciparum (SfR) 5.0%, respectively. P. falciparum was the dominant parasite accounting 51.2 per cent of the total infections. All the villages are situated in the deep forest and forest fringes. A total of 416 blood slides were collected and examined. Out of which 41 slides were found positive for malaria parasite (21 positive for P. falciparum and 20 positive for P. vivax). All the positive cases were treated with Blister Packs of anti-malarial of chloroquine and primaquine as per NBVDCP schedule to prevent further transmission of malaria, which were available. Rapid diagnostic Kits were used selectively only on the recommendation of the Physician/Medical Officer. Results of house to house fever survey indicated the presence of high percentage of symptomatic carriers of malaria parasites in the local population and in our study Pf % is more than reported data by District Malaria Office, Godda district, Jharkhand. In entomological studies, for mosquito fauna with reference to both Anopheline, 11 species belonging to one genus Anopheles were collected and identified during the survey; among them some species were most prevalent. A total of 599 mosquitoes were recorded and average density of mosquitoes recorded was 13.19 in human dwellings and 86.11 in cattle sheds.

Population structure and geographical subdivision of the Leishmania major vector Phlebotomus papatasi as revealed by microsatellite variation.

Multi-locus microsatellite typing (MLMT) has been employed to infer the population structure of Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) sandflies and assign individuals to populations. Phlebotomus papatasi sandflies were collected from 35 sites in 15 countries. A total of 188 P. papatasi individuals were typed using five microsatellite loci, resulting in 113 different genotypes. Unique microsatellite signatures were observed for some of the populations analysed. Comparable results were obtained when the data were analysed with Bayesian model and distance-based methods. Bayesian statistic-based analyses split the dataset into two distinct genetic clusters, A and B, with further substructuring within each. Population A consisted of five subpopulations representing large numbers of alleles that were correlated with the geographical origins of the sandflies. Cluster B comprised individuals collected in the Middle East and the northern Mediterranean area. The subpopulations B1 and B2 did not, however, show any further correlation to geographical origin. The genetic differentiation between subpopulations was supported by F statistics showing statistically significant (Bonferroni-corrected P < 0.005) values of 0.221 between B2 and B1 and 0.816 between A5 and A4. Identification of the genetic structure of P. papatasi populations is important for understanding the patterns of dispersal of this species and to developing strategies for sandfly control.

Development, malaria and adaptation to climate change: a case study from India.

India has reasons to be concerned about climate change. Over 650 million people depend on climate-sensitive sectors, such as rain-fed agriculture and forestry, for livelihood and over 973 million people are exposed to vector borne malarial parasites. Projection of climatic factors indicates a wider exposure to malaria for the Indian population in the future. If precautionary measures are not taken and development processes are not managed properly some developmental activities, such as hydro-electric dams and irrigation canal systems, may also exacerbate breeding grounds for malaria. This article integrates climate change and developmental variables in articulating a framework for integrated impact assessment and adaptation responses, with malaria incidence in India as a case study. The climate change variables include temperature, rainfall, humidity, extreme events, and other secondary variables. Development variables are income levels, institutional mechanisms to implement preventive measures, infrastructure development that could promote malarial breeding grounds, and other policies. The case study indicates that sustainable development variables may sometimes reduce the adverse impacts on the system due to climate change alone, while it may sometimes also exacerbate these impacts if the development variables are not managed well and therefore they produce a negative impact on the system. The study concludes that well crafted and well managed developmental policies could result in enhanced resilience of communities and systems, and lower health impacts due to climate change.

Studies on mosquito larvicidal properties of Eucalyptus citriodora Hook (family-Myrtaceae).

Hexane extract obtained from leaves of Eucalyptus citriodora Hook was tested against larvae of Anopheles stephensi Liston, Culex quinquefasciatus Say and Aedes aegypti Linn to assess its toxicity and growth inhibiting activity. The results showed better activity of hexane extract against larvae of An. stephensi Liston as compared to the larvae of Cx. quinquefasciatus Say and Ae. aegypti Linn. The LC50 values against IVth instar larvae of three species were 69.86, 81.12 & 91.76 ppm respectively after 24 hours and 26.7, 29.9 & 38.8 ppm respectively after 72 hours. At lowest concentration viz. 10 ppm, 73% larvae of An. stephensi Liston failed to emergence as adult mosquito while in Cx. quinquefasciatus Say and Ae. aegypti Linn only 10 & 6% larvae failed to emerge. Further observations showed some morphological abnormalities in same pupae which could not moult normally. These results clearly indicated growth inhibition effect of the hexane extract of Eucalyptus against mosquito larvae, particularly Anopheles species. Thus Eucalyptus citriodora Hook, which is commonly found in Northern India, has a great potential for the development of an eco-friendly herbal insecticidal product for control of mosquito breeding.

Laboratory study on larvicidal properties of leaf extract of Calotropis procera (Family-Asclepiadaceae) against mosquito larvae.

Fresh leaf extract of milkweed (Calotropis procera) showed larvicidal properties against mosquito larvae of Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. Methanolic extracts of the same plant were, however, more effective as larvicide. C. procera is a weed growing in uncultivated soils as well as in dry, arid zones in Indian subcontinent throughout the year. The results of this study suggest the utility of milkweed as potential technology for control of mosquito larvae.

Epidemiology of kala-azar in rural Bihar (India) using village as a component unit of study.

It is assumed that some features like intra-macrophage habitat of the Leishmania parasites, rare availability of infected macrophages in peripheral blood for vector sandflies to suck in, short flight range of sandflies, non-availability of an animal reservoir encountered in visceral leishmaniasis indicate slow and limited transmission potential and even so epidemics occur every 15 to 20 yr. To verify if these assumptions are true, the natural history of kala-azar was studied, using an endemic village in Bihar (India) as an unit of study, over a period of 5 yr (1984-1988). Village Jethuli is bound by the river Ganga on the north and separated from neighbouring endemic villages on other three sides by agricultural land, is isolated entomologically (as regards sandflies). The village has a population of 3236 persons of different social status and depending on economic conditions have three types of dwellings, brick made with cement plaster, brick made with mud plaster and mud houses. The first case of kala-azar was reported in a migrant from district Vaishali on the other side of Ganga where kala-azar appeared in an epidemic form. Studies showed that the infection is built up slowly, first in the same house and then in the immediate neighbourhood. In this village, maximum number of cases occurred in 1984 and 1985, and they were treated by our Institute and cured. In subsequent years, only a few cases occurred (i.e., 6 in 1986 and 4 in 1987) while no case occurred in 1988.(ABSTRACT TRUNCATED AT 250 WORDS)