Anopheles Fluviatilis Research Articles

Updated checklist with new records and molecular data for the mosquitoes (Diptera: Culicidae) of Hong Kong.

An extensive mosquito survey was carried out in Hong Kong from September to October 2022, employing a variety of collection methods. Specimens were identified using a combination of morphology and mitochondrial cytochrome C oxidase subunit 1 (COI) barcode sequences. Twenty-nine species, including three new records, i.e., Culex bicornutus (Theobald), Culex cinctellus Edwards, and Lutzia chiangmaiensis Somboon & Harbach, were collected. Phylogenetic analysis of COI sequences of Culex annulus Theobald and Culex vishnui Theobald collected in Hong Kong and elsewhere revealed that the sequences of the two nominal species are genetically very similar and are included in the same clade. Consequently, the synonymy of Cx. annulus with Cx. vishnui is reinstated. Lutzia halifaxii (Theobald) is removed from the list of species in Hong Kong and is replaced with Lutzia vorax Edwards, the identification of which is confirmed in the present study. The record of Culex spiculothorax Bram recorded in Hong Kong is replaced with the senior synonym Culex sasai Kano, Nitahara & Awaya. The occurrence of Anopheles fluviatilis James and Aedes aegypti (Linnaeus) is discussed. Finally, an updated checklist of the mosquitoes of Hong Kong, which now includes 76 species representing 14 genera, is provided, with notation of those species that vector pathogens of human diseases.

Population genetic structure of the malaria vector Anopheles fluviatilis species T (Diptera: Culicidae) in India.

Anopheles fluviatilis James (Diptera: Culicidae) represents a complex that comprises four sibling species (S, T, U, and V). Among these, species T is widely distributed in India. Chromosomal inversion polymorphism exists among different geographic populations of An. fluviatilis species T; however, population genetic structure is not understood. This study inferred a genetic structure among six geographically diverse populations of species T using a panel of microsatellite markers. Analyses indicated a significant but low genetic differentiation among the majority of the studied populations. A significant correlation was observed between genetic and geographic distances, exhibiting stepwise migration patterns among populations.

Malaria vector sibling species distribution in different endemic districts of Punjab, India.

Malaria transmission in Punjab, India is mainly seasonal with variation in its endemicity that may be due to varying vector behaviour in different areas of the state, primarily attributed to the existence of sibling species complexes among the vector species. So far there is no report regarding the existence of malaria vectors sibling species in the state of Punjab, therefore, the present study was planned to investigate the status of sibling species of two main vectors of malaria viz. Anopheles culcifacies and Anopheles fluviatilis in different districts of Punjab. Mosquito collections were made through hand catch in the morning hours. Malaria vector species An. culicifacies and An. fluviatilis were morphologically identified and man hour density was calculated. Both the vector species were subjected to molecular assays for sibling species identification through amplification of D3 domain of 28S ribosomal DNA by allele-specific PCR. Four sibling species of An. culicifacies, were identified viz. A, B, C and E. Species A was identified from Bhatinda district, species B, C and E from. S.A.S. Nagar and species C from Hoshiarpur. Two sibling species S and T of An. fluviatilis were identified from districts S.A.S. Nagar and Rupnagar. Presence of four sibling species of An. culicifacies and two sibling species of An. fluviatilis in Punjab necessitates planning of longitudinal studies to ascertain their role in disease transmission so that appropriate interventions may be applied to achieve malaria elimination.

Are members of the Anopheles fluviatilis complex conspecific?

Anopheles fluviatilis sensu lato, a primary malaria vector in India, has been identified to be comprised of four cryptic species, provisionally designated as species S, T, U and V. However, Kumar et al. (Mol Ecol Resour, 2013;13:354-61) considered all of the then known three members of this species complex (S, T and U) conspecific. The specific status of species S and T was refuted based on the lack of sufficient barcode gap in mitochondrial-CO1 and the perceived presence of heterozygotes in populations as detected through one of the two species-specific PCR assays employed for the cryptic species identification. The existence of species U was refuted claiming that earlier investigations have already refuted their existence. Here we discuss problems associated with the CO1-based barcode approach for delimitation of cryptic species, the perceived heterozygosity between species S and T based on a species-specific PCR assay, and interpretation of published reports. We demonstrated that fixed differences do exist in the ITS2-rDNA sequence of species S and T with no evidence of heterozygotes in sympatric populations and, that the observed heterozygosity by Kumar et al. in the ITS2-based species diagnostic PCR is due to the high mispriming tendency of the T-specific primer with species S. We infer that mitochondrial DNA-based ‘barcoding gap’, an arbitrary threshold recommended for species delimitation, alone, is inadequate to delimit the members of An. fluviatilis complex.

Molecular characterization of Anopheles fluviatilis species complex in the Islamic Republic of Iran

A species-specific polymerase chain reaction (PCR) assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran.

Population Dynamics and Insecticide Susceptibility of Anopheles culicifacies in Malaria Endemic Districts of Chhattisgarh, India.

Simple SummaryMalaria is a complex disease in part due to multiple vectors having different biological characteristics. In India, there are six primary vectors of malaria viz., Anopheles culicifacies, An. fluviatitlis, An. stephensi, An. subpictus, An. Minimus, and An. epiroticus. All these vectors have different ecological and seasonal distributions, transmission potential, and insecticide susceptibility status. In addition, except An. stephensi, all the five vectors comprise species complexes having sibling species which again differ in characteristics. Therefore, it is imperative to know the characteristics of the local vector population when it comes to planning control strategies. We carried out a study in tribal areas of Chhattisgarh state to investigate the seasonal and ecotype-wise distribution, breeding habits, sibling species composition, insecticide susceptibility, and role in the transmission of the local vector population. A high diversity of species was observed with around 16 species of Anopheles. An. culicifacies was the most dominant species and also was found to play a role in malaria transmission. The species was found to be resistant to dichlorodiphenyltrichloroethane (DDT) and Malathion, while an increasing trend of pyrethroid resistance was observed at some sites. Overall, our findings provide a picture of the characteristics of the local vector population in malaria-endemic regions.A study was undertaken in the villages of Korea and Bastar district (Chhattisgarh) during the years 2012–2015 to investigate the bionomics of malaria vectors and the prevalence of their sibling species complexes. Entomological surveys carried out every month included indoor resting collections, pyrethrum spray catches, light trap catches, and insecticide susceptibility status of Anopheles culicifacies using World Health Organization (WHO) methods. Anopheles culicifacies and Anopheles fluviatilis species were assayed by polymerase chain reaction (PCR) for the detection of malaria parasite, and sibling species were identified using PCR and DNA sequencing. A total of 13,186 samples of Anopheles comprising 15 species from Bastar and 16 from Korea were collected. An. Culicifacies was recorded as the most dominant species and also the only active vector at both sites. This species was found to be resistant to dichlorodiphenyltrichloroethane (DDT) and Malathion, showing signs of emerging resistance against pyrethroids. Among the sibling species of An. culicifacies, the group BCE was found in maximum numbers, while sibling species T of the An. fluviatilis was recorded to be dominant among its complex. The study provides a comprehensive view of the vector bionomics in the highly malarious regions of India that may have importance in developing vector control strategies.

Ecology of Malaria Vectors in an Endemic Area, Southeast of Iran.

Background:Malaria has long been regarded as one of the most important public health issues in Iran. Although the country is now in the elimination phase, some endemic foci of malaria are still present in the southeastern areas of the country. In some endemic foci, there are no data on the malaria vectors. To fill this gap, the present study was designed to provide basic entomological data on malaria vectors in the southeastern areas of Iran.Methods:Adult and larval stages of Anopheles mosquitoes were collected by using different catch methods. Resistance of the main malaria vector in the study area to selected insecticides was evaluated using diagnostic doses advised by the World Health Organization in 2013–2014.Results:A total of 3288 larvae and 1055 adult Anopheles mosquitoes were collected, and identified as: Anopheles stephensi (32.1%), Anopheles culicifacies s.l. (23.4%), Anopheles dthali (23.2%), Anopheles superpictus s.l. (12.7%), and Anopheles fluviatilis s.l. (8.6%). Anopheles stephensi was the most predominant mosquito species collected indoors at the study area, with two peaks of activity in May and November. This species was found to be resistant to DDT 4%, tolerant to malathion 5% and susceptible to other tested insecticides.Conclusion:All the five malaria vectors endemic to the south of Iran were collected and identified in the study area. Our findings on the ecology and resting/feeding habitats of these malaria vectors provide information useful for planning vector control program in this malarious area.

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

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. 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 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. 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.

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. 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. 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. 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.

A study of malaria vector surveillance as part of the Malaria Elimination Demonstration Project in Mandla, Madhya Pradesh

BackgroundUnderstanding of malaria vector density, distribution, insecticide resistance, vector incrimination, infection status, and identification of sibling species are some of the essential components of vector control measures for achieving malaria elimination goals.MethodsAs part of the malaria elimination demonstration project, entomological surveillance was carried out from October 2017 to October 2019 by collecting indoor resting mosquitoes using hand catch method. Susceptibility test was done for determining the insecticide resistance status of vector mosquito Anopheles culicifacies using standard protocols by the World Health Organization. The cone bioassay method was used for determining the efficacy and quality of insecticide sprayed. Mosquitoes collected from different ecotypes were identified and processed for parasite identification, vector incrimination and sibling species determination.ResultsThe two known malaria vector species (Anopheles culicifacies and Anopheles fluviatilis) were found in the study area, which have been previously reported in this and adjoining areas of the State of Madhya Pradesh. The prevalence of An. culicifacies was significantly higher in all study villages with peak in July while lowest number was recorded in May. Proportion of vector density was observed to be low in foothill terrains. The other anopheline species viz, Anopheles subpictus, Anopheles annularis, Anopheles vagus, Anopheles splendidus, Anopheles pallidus, Anopheles nigerrimus and Anopheles barbirostris were also recorded in the study area, although their prevalence was significantly less compared to the An. culicifacies. In 2017, An. culicifacies was found to be resistant to dichloro-diphenyl-trichloroethane (DDT) and malathion, with possible resistance to alphacypermethrin and susceptible to deltamethrin. However, in 2019, the species was found to be resistant to alphacypermethrin, DDT, malathion, with possible resistance to deltamethrin. The bioassays revealed 82 to > 98% corrected % mortality of An. culicifacies on day-one post-spraying and 35 to 62% on follow-up day-30. Anopheles culicifacies sibling species C was most prevalent (38.5%) followed by A/D and E while B was least pre-dominant (11.9%). Anopheles fluviatilis sibling species T was most prevalent (74.6%) followed by U (25.4%) while species S was not recorded. One An.culicifacies (sibling species C) was found positive for Plasmodium falciparum by PCR tests in the mosquitoes sampled from the test areas.ConclusionBased on the nine entomologic investigations conducted between 2017–2019, it was concluded that An. culicifacies was present throughout the year while An. fluviatilis had seasonal presence in the study areas. Anopheles culicifacies was resistant to alphacypermethrin and emerging resistance to deltamethrin was observed in this area. Anopheles culicifacies was confirmed as the malaria vector. This type of information on indigenous malaria vectors and insecticide resistance is important in implementation of vector control through indoor residual spraying (IRS) and use of insecticide-impregnated bed nets for achieving the malaria elimination goals.

Biting and resting behaviour of malaria vectors in Bandar-Abbas County, Islamic Republic of Iran.

Blood feeding and resting behaviour of malaria vectors are the most influential factors in malaria transmission. To measure blood feeding and resting behaviour, conventional mosquito sampling methods were performed in an area with potential for malaria transmission. Adult mosquitoes were collected monthly from indoor/outdoor places by conventional sampling methods, and larval habitats were investigated. Enzyme-linked immunosorbent assay was used to measure the human blood index (HBI) of the mosquitoes. Anopheles stephensi, Anopheles culicifacies, Anopheles dthali and Anopheles fluviatilis, were collected. Overall, 1249 female Anopheles mosquitoes were captured on human and animal baits, but no human-vector contact occurred indoors. A. dthali, A. fluviatilis and A. culicifacies showed a greater tendency to outdoor resting places in contrast to A. stephensi, which had a propensity to indoor resting places. The seasonal biting activities of all species occurred at average temperatures between 23 and 27°C. HBI was measured as 27.2%, 20.7%, 19.1%, and 23.0% for A. fluviatilis, A. stephensi, A. culicifacies and A. dthali, respectively. Vector control strategy depends upon mosquito behaviour. Therefore, using appropriate sampling methods based on mosquito behaviour is critical for malaria control planning. Exophilic/exophagic habit of mosquito vectors leads to fewer human bites, resulting in biting protection. Exophilic behaviour also requires specific larvicidal operations in order to prevent and control malaria transmission.

Modified PCR-based assay for the differentiation of members of Anopheles fluviatilis complex in consequence of the discovery of a new cryptic species (species V)

BackgroundAnopheles fluviatilis is a species-complex comprising of four cryptic species provisionally designated as species S, T, U and V. Earlier, a 28S-rDNA based allele-specific polymerase chain reaction (ASPCR) assay was developed for the differentiation of the then known three members of the An. fluviatilis complex, i.e., species S, T, and U. This assay was modified in consequence of the discovery of a new cryptic member, species V, in the Fluviatilis Complex to include identification of new species.MethodsIn the modified procedure, the ASPCR assay was performed first, followed by restriction digestion of PCR product with an enzyme BamH I, which cleaves specifically PCR amplicon of species V and the resultant PCR–RFLP products can differentiate all the four cryptic members of the complex. Morphologically identified An. fluviatilis samples were subjected to sibling species identification by modified PCR-based assay and standard cytotaxonomy. The result of PCR-based assay was validated through cytotaxonomy as well as DNA sequencing of some representative samples.ResultsThe modified PCR-based assay differentiates all four sibling species. The result of modified PCR-based assay tested on field samples was in agreement with results of cytotaxonomy as well as DNA sequencing of representative samples.ConclusionsThe modified PCR-based assay unambiguously differentiates all four known members of the An. fluviatilis species complex. This assay will be useful in studies related to bionomics of members of the Fluviatilis Complex in their role in malaria transmission.

Variations in palpal ornamentation of Anopheles fluviatilis species T and U (Diptera: Culicidae) and their taxonomic consequence

Anopheles fluviatilis sensu lato James is a highly efficient malaria vector in Indian subcontinent and Iran which is comprised of at least four sibling species provisionally designated as species S, T, U and V. An important morphological characteristic for differentiation of this species complex from other closely related anopheline species complex, the Minimus Complex, is the ratio of length of subapical pale band to dark band intervening apical and subapical pale bands on the maxillary palps of female mosquito. Here, we report variation in the subapical pale band in An. fluviatilis , especially in species U, to the extent that palps of some specimens resemble members of Minimus Complex, inferring that palpal ornamentation may not be reliable characteristics for identification of An. fluviatilis . Taxonomic consequence of such variation is discussed

Climate Change and the Risk of Malaria Transmission in Iran

Climate change is an important factor affecting the dynamics of the vectors population and, hence, the risk of vector-borne diseases. This study aimed to predict the environmental suitability for malaria vectors in Iran under climate change scenarios in 2030s and 2050s. Literature search was performed to find documents on the spatial distribution of Anopheles stephensi Liston, 1901, Anopheles culicifacies s.l. Giles, 1901, Anopheles fluviatilis s.l. James, 1902, Anopheles superpictus s.l. Grassi, 1899, Anopheles dthali Patton, 1905, Anopheles maculipennis s.l. Meigen, 1818, and Anopheles sacharovi Favre, 1903 (Diptera: Culicidae) published between 1970 and 2017. The bioclimatic data under three climate change scenarios (representative concentration pathway 2.6 [RCP2.6], RCP4.5, and RCP8.5) and MaxEnt model were used to predict the ecological niches for each species. Comparison between the two study periods under the three scenarios for each species revealed that RCP8.5 would reduce the area at risk for An. culicifacies s.l., An. dthali and An. superpictus s.l. in the 2050s compared to the 2030s, but the reverse will be induced by RCP2.6 and RCP4.5 scenarios. For An. fluviatilis s.l., RCP2.6 will reduce the risk areas in the 2050s, whereas an increase is expected under the two other scenarios. Moreover, all scenarios would decrease the high-risk areas of An. maculipennis s.l. in the 2050s. For An. sacharovi, RCP2.6 would increase its high-risk areas, whereas RCP4.5 and RCP8.5 would decrease its exposure. The high-risk area of An. stephensi is expected to increase under RCP8.5 in the 2030s and RCP4.5 in 2050s, but it will be almost unchanged or reduced under other scenarios.

Species composition, seasonal abundance and distribution of potential anopheline vectors in a malaria endemic area of Iran: field assessment for malaria elimination

BackgroundDespite decreases in incidence, malaria remains a major public health challenge in south and southeast Iran, where vector control is considered one of the main strategies for elimination of the disease. The efficacy of this strategy depends on understanding malaria vector ecology, which varies by species. This study was conducted to determine the species composition, seasonal abundance and distribution of potential anopheline vectors in Bashagard County, one of the important malaria-endemic areas in south Iran.MethodsIn this cross-sectional study, four typical foothill and mountainous villages were selected by simple random sampling. Anopheline mosquitoes were collected by the standard dipping method for larvae and total catch for adults. Anopheline specimens were morphologically identified using taxonomic keys. Statistical analyses were performed using SPSS ver.20 software.ResultsIn total, 1211 anopheline specimens, including 1055 (87.12%) larvae and 156 (12.88%) adults, were collected and identified. They consisted of 9 species: Anopheles moghulensis (27.89%), Anopheles dthali (18.91%), Anopheles culicifacies (16.60%), Anopheles stephensi (15.38%), Anopheles turkhudi (9.83%), Anopheles superpictus (9.66%), Anopheles apoci (1.40%), Anopheles fluviatilis (0.17%), and Anopheles sergentii (0.08%). The most prevalent species in adult catches were An. culicifacies (44.23%), An. dthali (21.15%) and An. stephensi (16.03%), and the most prevalent species caught as larvae were An. moghulensis (31.94%), An. dthali (18.85%) and An. stephensi (15.26%). Most of the anopheline species were distributed in different topographical areas and two proven malaria vectors, An. culicifacies and An. superpictus, were significantly associated with altitude and collected more frequently in the foothill regions. Most of the anopheline species were present almost throughout the year with a major peak in April and a smaller peak in October.ConclusionThe results of this study revealed that there are five malaria vectors in Bashagard County and some of them are more abundant in areas with special topographic features and are reproductively active throughout the year. These findings can be applied to successful planning and providing effective control measures in problematic areas during the malaria elimination phase in Iran.

Comparative efficacy of two rounds of indoor residual spraying of DDT 75% @ 1g/m2 with that of DDT 50% @ 1g/m2 against the malaria vectors in India

While, dichlorodiphenyl-trichloroethane (DDT) water dispersible powder (WDP) 75% is considered as the high performance long lasting formulation for indoor residual spraying (IRS), no information is available regarding the comparative epidemiological effectiveness of the two DDT formulations when used for IRS in Indian conditions. The current study was undertaken to compare the effectiveness of IRS using DDT WDP 75% @ one g active ingredient (AI)/m2 with that using DDT WDP 50% @ one g AI/m2 in controlling Anopheles fluviatilis and An. culicifacies, the primary vectors of malaria in the selected endemic areas of Odisha State. Although, cone-bioassay mortality after 8 months of post spraying on DDT 75% sprayed surfaces as well as on sprayed but mud plastered surfaces was higher than DDT 50%, the six entomological parameters viz. resting density indoors and outdoors, trap density indoors, parous rate, human blood index and infection rate of An. fluviatilis and An. culicifacies did not show any statistically significant difference in reduction/ changes from pre- to post-spray period between the two DDT formulations.

Evaluation of MAGNet, a long-lasting insecticidal mosquito net against Anopheles fluviatilis in experimental huts in India

BackgroundMAGNet LN is a wash resistant long-lasting insecticidal (polyethylene) net (LLIN) in which the alpha-cypermethrin insecticide is incorporated within the 150 denier high density polyethylene monofilaments of the nets. The bio-efficacy of MAGNet LN was reported to be high even after 25 washes. The LN met the WHO criteria of Phase I evaluation and obtained recommendation from the World Health Organization Pesticide Evaluation Scheme (WHOPES) for Phase II trial. For registration of the LN in India, the current study was conducted to evaluate its efficacy after 20 or 25 washes compared to negative control (untreated net) and positive control (Duranet LN) in experimental huts against a wild, free flying pyrethroid susceptible population of Anopheles fluviatilis in terms of deterrence, blood-feeding inhibition, mortality and induced exophily.MethodsThe evaluation was carried out in six experimental huts located at Kandhaguda village in Malkangiri district, Odisha state following the WHO guidelines.ResultsThe study showed that 25 times washed MAGNet LN produced 100% mortality in cone bioassays before and after hut evaluation. MAGNet washed 25 times did not differ significantly from all other treated nets in terms of deterring hut entry, induced exophily, blood feeding inhibition and causing mortality of An. fluviatilis.ConclusionsMAGNet LN showed extended wash resistance retaining its bio-efficacy up to 25 washes and met the WHOPES requirement of passing Phase II evaluation.

Bio-efficacy of LifeNet, a deltamethrin incorporated long-lasting insecticidal net, as assessed in experimental huts against Anopheles fluviatilis, a major malaria vector in east-central India

LifeNet, a deltamethrin incorporated long-lasting insecticidal (polypropylene) net (LLIN), was qualified by the World Health Organization Pesticide Evaluation Scheme (WHOPES) for Phase-II trial in India. The purpose of this trial was to assess the bio-efficacy of unwashed and 20 and 30 times washed LifeNet in comparison to the nets conventionally treated with deltamethrin against the natural population of Anopheles fluviatilis, a major malaria vector, in terms of deterring hut entry, inhibiting blood feeding, inducing exophily and causing mortality. The trial was carried out in six experimental huts constructed at Kandhaguda village in Malkangiri district, Odisha State. The efficacy of unwashed and washed (20 or 30 times) LifeNet was compared with untreated polypropylene and conventionally treated (with deltamethrin) polyester net washed to just before exhaustion or washed 20 times. The study showed a significant reduction of entry (treatment: 1.61–4.78; control: 7.61 per hut) and an increase in exit (50.7–64.4% and 39.1%) of An. fluviatilis in the treated arms compared to the control arm (untreated net) (P < 0.05). Blood feeding rates reduced in treated arms (20.7–68.0%) compared to the control (80.3%) (P < 0.05). Total mortality was significantly higher in LifeNet arms (73.8–98.3%) than the control (2.2%) (P < 0.05). After 30 washes, the active ingredient (AI) retention in LifeNet was 62%. Performance of the three LifeNet arms against the susceptible population of An. fluviatilis met the WHO efficacy criteria of Phase II evaluation for LLINs.

Malaria epidemiology in an area of stable transmission in tribal population of Jharkhand, India

BackgroundMalaria remains an important health problem in India with approximately 1 million cases in 2014. Of these, 7% occurred in the Jharkhand state mainly in the tribal population.MethodsThis study was conducted in Dumargarhi, a tribal village about 42 km east of Ranchi city, Jharkhand, from May 2014 to September 2016. Four point prevalence surveys were carried out during consecutive high (October–December) and low (June–August) transmission seasons. Malaria cases were recorded from April 2015 to April 2016 through fortnightly visits to the village. Adult mosquito densities were monitored fortnightly by manual catching using suction tube method.ResultsThe study area consists of five hamlets inhabited by 945 individuals living in 164 households as recorded through a house-to-house census survey performed at enrollment. The study population consisted predominantly of the Munda (n = 425, 45%) and Oraon (n = 217, 23%) ethnic groups. Study participants were categorized as per their age 0–5, 6–10, 11–15 and >15 years. There were 99 cases of clinical malaria from April 2015 to April 2016 and all malaria cases confirmed by microscopy were attributed to Plasmodium falciparum (94 cases) and Plasmodium vivax (5 cases), respectively. During the high transmission season the mean density of P. falciparum parasitaemia per age group increased to a peak level of 23,601 parasites/μl in the 6–10 years age group and gradually declined in the adult population. Malaria attack rates, parasite prevalence and density levels in the study population showed a gradual decrease with increasing age. This finding is consistent with the phenomenon of naturally acquired immunity against malaria. Three vector species were detected: Anopheles fluviatilis, Anopheles annularis, and Anopheles culicifacies. The incoherence or complete out of phase pattern of the vector density peaks together with a high prevalence of parasite positive individuals in the study population explains the year-round malaria transmission in the study region.ConclusionsThe collection of clinical data from a well-characterized tribal cohort from Jharkhand, India, has provided evidence for naturally acquired immunity against malaria in this hyperendemic region. The study also suggests that enforcement of existing control programmes can reduce the malaria burden further.

Bionomics of Anopheles fluviatilis and Anopheles culicifacies (Diptera: Culicidae) in Relation to Malaria Transmission in East-Central India.

The southern districts of Odisha State in east-central India have been highly endemic for falciparum malaria for many decades. However, there is no adequate information on the abundance of the vector species or their bionomics in relation to space and time in these districts. Therefore, a study was carried out on the entomological aspects of malaria transmission to generate such information. Collections of mosquitoes were made once during each of the three seasons in 128 villages selected from eight districts. Villages within the foot-hill ecotype had a significantly greater abundance of Anopheles fluviatilis James s. l., whereas the abundance of Anopheles culicifacies Giles s. l. was significantly greater in the plain ecotype. The abundance of An. fluviatilis was maximum during the cold season, whereas An. culicifacies abundance was highest during summer and rainy seasons. The maximum likelihood estimation of the malaria infection rate in An. fluviatilis was 1.78%, 6.05%, and 2.6% in Ganjam, Kalahandi, and Rayagada districts, respectively. The infection rate of An. culicifacies was 1.39% only in Kandhamal district; infected females were not detected elsewhere. Concurrently, the annual malaria parasite incidence (MPI) was significantly higher in hill-top (17.6) and foot-hill (14.4) villages compared to plain villages (4.1). The districts with more villages in hill-top and foot-hill ecotypes also had a greater abundance of An. fluviatilis, the major malaria vector, and exhibited a higher incidence of malaria than villages within the plain ecotype, where An. culicifacies was the most abundant vector.