Isolation and characterization of emerging Mpox virus from India.

Mpox (previously known as Monkeypox) has recently re-emerged, primarily through human-to-human transmission in non-endemic countries including India. Virus isolation is still considered as the gold standard for diagnosis of viral infections. Here, the qPCR positive skin lesion sample from a patient was inoculated in Vero E6 cell monolayer. Characteristic cytopathic effect exhibiting typical cell rounding and detachment was observed at passage-02. The virus isolation was confirmed by qPCR. The replication kinetics of the isolate was determined that revealed maximum viral titre of log 6.3 PFU/mL at 72 h postinfection. Further, whole genome analysis through next generation sequencing revealed that the Mpox virus (MPXV) isolate is characterized by several unique SNPs and INDELs. Phylogenetically, it belonged to A.2 lineage of clade IIb, forming a close group with all other Indian MPXV along with few from USA, UK, Portugal, Thailand and Nigeria. This study reports the first successful isolation and phenotypic and genotypic characterization of MPXV from India.

Effects of weak static magnetic fields on the development of seedlings of Arabidopsis thaliana.

To study magnetoreception of Arabidopsis thaliana, we analysed several developmental responses including cryptochrome-independent seed germination and the phytochrome- and cryptochrome-dependent hypocotyl elongation and photo-accumulation of anthocyanins and chlorophylls in weak static magnetic fields ranging from near null to 122 µT. A field of 50 µT accelerated seed germination by about 20 h relative to samples maintained in a near-null field. The double mutant, cry1cry2, lacking cryptochromes 1 and 2 displayed the same magnetic field-induced germination acceleration under blue light as the wild-type strain. Magnetic field-induced germination acceleration was masked in the presence of exogenous sucrose. Stimulus-response curves for hypocotyl elongation in a range between near-null to 122 µT indicated maxima near 9 and 60 µT for the wild-type strain as well as mutant cry1cry2. The photo-accumulation of anthocyanins and chlorophylls could be effectively modulated by magnetic fields in the presence of low-irradiance red and blue light, respectively. The findings indicate that Arabidopsis thaliana possesses light-independent mechanisms of magnetic field reception, which remain presently unidentified. Our results are in better agreement with predictions of the level crossing mechanism (LCM) of magnetoreception rather than those of the cryptochrome-associated radical-pair mechanism (RPM).

Harnessing heterosis and male sterility in soybean [Glycine max (L.) Merrill]: A critical revisit.

Soybean is a predominantly self-pollinated crop. It is also one of the important oilseed legumes. Soybean is an excellent crop having industrial, traditional, culinary, feeding, and cultural roles. Genetic diversity in breeding programs is of prime importance as it ensures the success of any breeding by enhancing the outcomes and results of the plants. The phenomenon wherein the progeny exhibits greater biomass (yield) and a faster rate of development and fertility than its parents is referred to as heterosis. As of now, heterosis is mainly limited to the trait of seed yield and is considered the basis for the development of better (superior) varieties. Male sterility (MS) is extensively used for the production of seeds and the improvement of crops coupled with the traditional breeding programs and molecular technology. Therefore, deployment of MS and heterosis in breeding soybean could yield better outcomes. This review aims to focus on two aspects, namely, MS and heterosis in soybean with its scope for crop improvement.

Deltamethrin Contact Exposure Mediated Toxicity and Histopathological Aberrations in Tissue Systems of Public Health Importance Cockroach Species Periplaneta americana and Blattella germanica.

Cockroach species Periplaneta americana and Blattella germanica potentially survive in locations close to human activity. Besides spoiling food material, cockroaches also transfer pathogens of different diseases among human beings. Since the insecticides have been used extensively to control cockroaches, information on their insecticide susceptibility and toxicity at the cellular level may be crucial. In the study, deltamethrin toxicity as well as the deltamethrin-mediated cytomorphological changes in the brain, ovary and midgut of the two important cockroach species have been assessed. Different concentrations [0.00025% (0.0025 mg/ml), 0.0025% (0.025 mg/ml), 0.025% (0.25 mg/ml), 0.25% (2.5 mg/ml), 0.5% (5 mg/ml), 1% (10 mg/ml)] of deltamethrin in acetone were used to expose test species in WHO bottle assay. Knockdown was recorded after 5 min interval while delayed mortality was observed after 24 h. Brain, ovary and gut were dissected post 1 h exposure and 24 h holding (for 0.25, 0.5 and 1% concentration), and tissues were processed for microscopic analysis. Deltamethrin exposed cockroaches and dissected tissues were used to estimate deltamethrin using HPLC. At 0.00025% (lowest concentration), the percentage knock-down observed was 66.7% for P. americana and 80% B. germanica respectively (R 2 = 0.78; p = 0.0001) in 1 h. KDT50 value was found to be 8.7 min (95% CI: 7.3-10.2), while KDT99 was 20.7 min (95% CI: 16.0-35.7) in P. americana at 1% concentration. Whereas, the KDT50 and KDT99 values for B. germanica were 7.4 min (95% CI: 5.4-9.1) and 27.4 min (95% CI: 18.2-80.0) at a similar concentration. LD50 and LD95 values (for 60 min standard exposure) were 0.0006% (95% CI: 0.00-0.001) and 0.034% (95% CI: 0.013-0.49) respectively for P. americana, while these values were 0.0005 (95% CI: 0.00-0.001) and 0.04 (95% CI: 0.01-0.23) for B. germanica. Exposure to 1% deltamethrin induced a considerable toxic effect in the epithelial cells in the midgut. HPLC estimated 0.21 ± 0.05 mg (95% CI: 0.18-0.25; CoV 23.9%) deltamethrin in P. americana post 1% exposure. Even short term exposure to a low concentration of synthetic pyrethroid deltamethrin displayed immediate knockdown and delayed mortality in both the test species. Considerable histological damage was observed in both the insects at 1% exposure. In India, resistance to deltamethrin may have been reported among different insects due to its extensive use. However, the formulations such as insecticide paints, attractant baits etc. developed using deltamethrin as an active ingredient could be useful in cockroach control operations.

Deltamethrin Microencapsulation in Emulsion Paint Binder and Its Long-Term Efficacy Against Dengue Vector Aedes aegypti.

Various control interventions have been effective in the control of arthropod vectors to a certain extent; still, sustained vector control is an existing problem globally. Insecticide-based formulations have been found to be useful, however the proper delivery of active molecules to target vectors is important. Currently, synthetic pyrethroid deltamethrin (DM) has been microencapsulated in the emulsion paint binder and evaluated for long-term effectiveness against dengue vector Aedes aegypti. Different compositions of emulsion binder were prepared by varying the content of monomer and DM. A selection was made for the composition yielding the best combination of properties like solid content, intrinsic viscosity, and DM content. Developed formulation was tested against laboratory-reared and pathogen-free Ae. aegypti mosquitoes. Encapsulation of DM in emulsion binder during polymerization showed a uniform distribution. The optimized formulation was stable and did not have a considerable plasticizing effect. Scanning electron microscopy revealed that grain-like micro crystals of DM and surfactant sodium lauryl sulfate (SDS) were uniformly distributed on the formulation surface. The best optimized formulation was highly effective against dengue vector Ae. aegypti and found to provide efficacy for up to 18 months of application. The knockdown time (KDT) values KDT10 and KDT50 were 7.4 min (95% CI: 5.6-9.1) and 22.1 min (95% CI: 19.7-24.3) respectively, whereas 24 h corrected mortality was 90% (95% CI: 82.5-97.5) after 18 months of application (T18). The probit model used to determine knockdown values did not deviate from the linearity and displayed normal distribution of knockdown % with time for different formulations (p ≥ 0.1). Presently developed DM microencapsulated emulsion binder was stable, smooth, and uniform. The binder displayed excellent anti-insect property and was capable of providing long-term effectiveness against dengue vectors Ae. aegypti. Such a formulation after field-scale evaluation could be very useful in attaining long-term protection from arthropod vectors.

Behavioural response of mosquito vectors Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus to synthetic pyrethroid and organophosphorus-based slow-release insecticidal paint.

BACKGROUND: The direct toxicological impact of insecticides on vector mosquitoes has been well emphasized; however, behavioural responses such as excito-repellency and physical avoidance as a result of insecticide exposure have not been much studied. We have demonstrated the excito-repellency and behavioural avoidance in certain vector mosquito species on exposure to a slow-release insecticidal paint (SRIP) formulation in addition to direct toxicity. METHODS: A SRIP formulation developed by the Defence Research and Development Establishment, Gwalior, contains chlorpyriphos, deltamethrin and pyriproxyfen as active insecticides. Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti mosquitoes were used to study the excito-repellency response of the formulation. The experiments were performed in a specially designed dual-choice exposure and escape chamber made of transparent polymethyl methacrylate. For the experiments, the SRIP formulation was applied undiluted at a rate of 8 m2 per kg on 15 cm2 metallic surfaces. Mosquitoes were introduced into the exposure chamber, and observations of the movement of mosquitoes into the escape chamber through the exit portal were taken at 1-min intervals for up to 30 min. RESULTS: The evaluated formulation displayed strong excito-repellency against all three tested vector mosquito species. Results showed that the ET50 (escape time 50%) for Ae. aegypti, An. stephensi and Cx. quinquefasciatus was 20.9 min, 14.5 min and 17.9 min for contact exposure (CE) respectively. Altogether in CE, the escape rates were stronger in An. stephensi mosquitoes at different time intervals compared to Ae. aegypti and Cx. quinquefasciatus mosquitoes. The probit analysis revealed that the determined ET did not deviate from linearity for both non-contact exposure (NCE) and placebo exposure (PE) (χ2 ≤ 7.9; p = 1.0) for Ae. aegypti mosquitoes and for NCE (χ2 = 8.3; p = 1.0) and PE (χ2 = 1.7; p = 1.0) treatments in Cx. quinquefasciatus. Mortality (24 h) was found to be statistically higher (F = 6.4; p = 0.02) in An. stephensi for CE but did not vary for NCE (p ≥ 0.3) and PE (p = 0.6) treatments among the tested mosquito species. Survival probability response suggested that all the three tested species displayed similar survival responses for similar exposures (χ2 ≤ 2.3; p ≥ 0.1). CONCLUSION: The study demonstrates the toxicity and strong behavioural avoidance in known vector mosquito species on exposure to an insecticide-based paint formulation. The combination of insecticides in the present formulation will broaden the overall impact spectrum for protecting users from mosquito bites. The efficacy data generated in the study provide crucial information on the effectiveness of the tested formulation and could be useful in reducing the transmission intensity and disease risk in endemic countries.

Pyriproxyfen treated surface exposure exhibits reproductive disruption in dengue vector Aedes aegypti.

BACKGROUND: Reduced susceptibility of mosquito vectors to currently used insecticides hampers control interventions. Recently pyriproxyfen, an insect growth regulator has been demonstrated to effectively reduce the reproductive potential in vector mosquitoes. METHODS: Pyriproxyfen (PPF), in different concentrations (0.75%, 0.075%, 0.0075%, and 0.00075%) was applied on papers and Indian wild type Aedes aegypti female mosquitoes (N ≥ 20 for each treatment) were exposed onto it as per WHO guidelines, to study the reproductive disruption. PPF concentration on treated papers was quantitatively cross-determined using HPLC method. Reduction in fecundity, fertility and adult emergence in exposed female Ae. aegypti was determined. Abnormal development in ovary and eggs of exposed females was studied microscopically after different time intervals. RESULTS: Eggs laid, eggs hatched, pupae formed and adults emerged per female exposed in both before blood meal and after blood meal groups declined significantly from lowest to highest concentration of PPF (F ≥ 5.2; p < 0.02). Adult emergence inhibition in females exposed to PPF before and after blood meal groups ranged from 58.8% [OR = 0.18 (95% CI = 0.09-0.36)] to 79.2% [OR = 0.04 (95% CI = 0.02-0.10)] and 64.4% [OR = 0.12 (95% CI = 0.05-0.28)] to 77.1% [OR = 0.05 (95% CI = 0.02-0.14)] respectively in different concentrations. The probit model used suggested that FI50 (50% fertility inhibition) and EI50 (50% emergence inhibition) were 0.002% (p = 0.82) and 0.0001% (p = 0.99) for females exposed before blood meal, while 0.01% (p = 0.63) and <0.0001% (p = 0.98) for the females exposed after blood meal, respectively. The eggs laid by the females exposed to PPF-treated surface showed altered body organization, desegmentation and disoriented abdominal and cervical regions in the developing embryo. Quantification of PPF on impregnated papers showed that it was uniformly distributed throughout the matrix. CONCLUSIONS: The present study has shown that tarsal contact to PPF-treated surface for a small time drastically influenced the fecundity, fertility and adult emergence in Indian wild Ae. aegypti mosquitoes. Results suggest that a certain minimum concentration of PPF through contact exposure can reduce the abundance of vector mosquitoes to a considerable level. The formulations based on combination of PPF and other compatible insecticides may be an impactful approach where susceptible mosquitoes are killed by the insecticide component while resistant mosquitoes are sterilised by PPF.

Correction to: Are malaria elimination efforts on right track? An analysis of gains achieved and challenges ahead.

AbstractIn the original publication of this article [1], the legends in Fig. 9 are missing during the process of typesetting.

Are malaria elimination efforts on right track? An analysis of gains achieved and challenges ahead.

BACKGROUND: Malaria causes significant morbidity and mortality each year. In the past few years, the global malaria cases have been declining and many endemic countries are heading towards malaria elimination. Nevertheless, reducing the number of cases seems to be easy than sustained elimination. Therefore to achieve the objective of complete elimination and maintaining the elimination status, it is necessary to assess the gains made during the recent years. MAIN TEXT: With inclining global support and World Health Organisation (WHO) efforts, the control programmes have been implemented effectively in many endemic countries. Given the aroused interest and investments into malaria elimination programmes at global level, the ambitious goal of elimination appears feasible. Sustainable interventions have played a pivotal role in malaria contraction, however drug and insecticide resistance, social, demographic, cultural and behavioural beliefs and practices, and unreformed health infrastructure could drift back the progress attained so far. Ignoring such impeding factors coupled with certain region specific factors may jeopardise our ability to abide righteous track to achieve global elimination of malaria parasite. Although support beyond the territories is important, but well managed integrated vector management approach at regional and country level using scrupulously selected area specific interventions targeting both vector and parasite along with the community involvement is necessary. A brief incline in malaria during 2016 has raised fresh perturbation on whether elimination could be achieved on time or not. CONCLUSIONS: The intervention tools available currently can most likely reduce transmission but clearing of malaria epicentres from where the disease can flare up any time, is not possible without involving local population. Nevertheless maintaining zero malaria transmission and checks on malaria import in declared malaria free countries, and further speeding up of interventions to stop transmission in elimination countries is most desirable. Strong collaboration backed by adequate political and financial support among the countries with a common objective to eliminate malaria must be on top priority. The present review attempts to assess the progress gained in malaria elimination during the past few years and highlights some issues that could be important in successful malaria elimination.

Effects of weak static magnetic fields on the gene expression of seedlings of Arabidopsis thaliana.

Magnetic-field reception of animals and plants is currently discussed in the framework of a cryptochrome-based radical-pair mechanism. Efforts to unravel magnetoreception in plants suffered historically from several shortcomings, most prominently, the conspicuous absence of detailed stimulus-response relationships. To determine the sensitivity of seedlings of Arabidopsis thaliana to weak static magnetic fields we generated stimulus-response curves between near zero and 188 µT for the transcript levels of the genes rbcl, cab4, pal4 and ef1. The moderate magneto-responsiveness of dark-grown seedlings was greatly enhanced under blue light, and for rbcl and pal4 also under red light. The stimulus-response curves obtained under blue light of constant photon-fluence rate displayed multiple maxima and thus a pattern fundamentally different from that prevalent in plant and animal physiology. A double mutant lacking cryptochromes 1 and 2 displayed altered stimulus-response curves without losing, however, magneto-responsiveness completely. A reversal of the magnetic field direction substantially affected the gene expression and the quantity of CAB-protein (chlorophyll a,b-binding protein). The majority of our results are at variance with the notion of cryptochromes acting as the only magnetic-field sensors. They do not, however, exclude the possibility that cryptochromes participate in the magnetic field reception of Arabidopsis. The findings have the unexpected implication that cryptochrome- and phytochrome-mediated plant responses can be modulated by the strength and the orientation of the local geomagnetic field.

Disappearance of Anopheles minimus and Anopheles dirus from Certain Malaria Endemic Areas of Assam, India.

BACKGROUND: Orang Primary Health Centre (OPHC) and Balipara Primary Health Centre (BPHC) of Assam (India) report mosquito borne diseases annually. Current study was performed to ascertain the prevalence of known malaria and Japanese Encephalitis (JE) vectors and their possible role in disease transmission. METHODS: Malaria epidemiological data for 2006-2010 and JE data for 2008-2013 of Assam, India were obtained from the health authority. Mosquitoes were collected using CDC light traps and identified morpho-taxonomically. RESULTS: Plasmodium falciparum cases (81.5%, 95% CI= 72.0-91.1) were statistically higher in OPHC (P< 0.0001, t= 8.0) during the recent years. There was 4.4 folds rise in the confirmed acute encephalitis syndrome (AES) and 3.2 folds increase in the confirmed JE cases during 2013 as compared to 2008. Altogether 9,218 mosquito specimens (PTND= 153.6), comprising of 44.1% anophelines (PTND= 67.7), 42.3% culicines (PTND= 65.0) and 9.5% mansonia (PTND= 14.6) were recorded. In BPHC, Anopheles vagus was recorded in high density (P< 0.0001), whereas Culex quinquefasciatus was the predominant JE vector (P= 0.04). In OPHC, among the known malaria vectors, the density of Anopheles annularis was significantly high (P< 0.0001). However Culex bitaeniorhynchus was the predominant known JE vector (P< 0.0001) followed by Cx. quinquefasciatus. CONCLUSION: Even in the absence of known efficient vectors, many Anopheles species are still involved in malaria transmission. There was disappearance of Anopheles minimus and Anopheles dirus and establishment of An. annularis, An. vagus and An. philippinensis/nivipes mosquitoes in study area.

Morphometric and morphological appraisal of the eggs of Anopheles stephensi (Diptera: Culicidae) from India.

BACKGROUND & OBJECTIVES: Anopheles stephensi is one of the most important urban malaria vectors in India and contribute about 12% of total malaria cases. An. stephensi has three ecological variants; type, intermediate and mysorensis that can be differentiated on the basis of differences in number of ridges on egg float and on the basis of spiracular indices. Because of its anthropophilic nature the 'type' form is an efficient malaria vector. In the present study, the egg surface morphometry and morphology of An. stephensi 'type' form was studied and detail distinguish- ing characters were recorded for its correct identification. METHODS: Eggs of An. stephensi 'type' form were studied by scanning electron microscopy (SEM) after sputter- coating with gold. In total 23 egg characters were analysed morphologically and morphometrically, which included egg attributes, deck attributes, ventral tubercles, micropyle and float attributes. RESULTS: The dorsal surface of the egg of 'type' form was curved while the ventral surface was concave and both anterior and posterior ends were blunt. The average length and width of egg was 473.94 + 11.18 and 154.69 + 2.66 µm respectively. The number of float ribs observed was 20.33 ± 0.33. The maximum length of float was found to be 246.57 + 15.27 µm, whereas maximum width was 87.16 + 3.83 µm. INTERPRETATION & CONCLUSION: The present study has generated some important data which is specific to An. Stephensi 'type' form and provided significant morphological and morphometric standards for its correct identification. This information could be useful in differentiation of An. stephensi 'type' form from other ecological forms of the same species as well as other species of Anopheles.

Protection against mosquito vectors Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus using a novel insect repellent, ethyl anthranilate.

Growing concern on the application of synthetic mosquito repellents in the recent years has instigated the identification and development of better alternatives to control different mosquito-borne diseases. In view of above, present investigation evaluates the repellent activity of ethyl anthranilate (EA), a non-toxic, FDA approved volatile food additive against three known mosquito vectors namely, Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus under laboratory conditions following standard protocols. Three concentration levels (2%, 5% and 10% w/v) of EA were tested against all the three selected mosquito species employing K & D module and arm-in-cage method to determine the effective dose (ED50) and complete protection time (CPT), respectively. The repellent activity of EA was further investigated by modified arm-in-cage method to determine the protection over extended spatial ranges against all mosquito species. All behavioural situations were compared with the well-documented repellent N,N-diethylphenyl acetamide (DEPA) as a positive control. The findings demonstrated that EA exhibited significant repellent activity against all the three mosquitoes species. The ED50 values of EA, against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus were found to be 0.96%, 5.4% and 3.6% w/v, respectively. At the concentration of 10% w/v, it provided CPTs of 60, 60 and 30min, respectively, against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus mosquitoes. Again in spatial repellency evaluation, EA was found to be extremely effective in repelling all the three tested species of mosquitoes. Ethyl anthranilate provided comparable results to standard repellent DEPA during the study. Results have concluded that the currently evaluated chemical, EA has potential repellent activity against some well established mosquito vectors. The study emphasizes that repellent activity of EA could be exploited for developing effective, eco-friendly, acceptable and safer alternative to the existing harmful repellents for personal protection against different hematophagous mosquito species.

Malaria vector Anopheles culicifacies sibling species differentiation using egg morphometry and morphology.

BACKGROUND: The malaria vector Anopheles culicifacies (sensu lato) is an important malaria vector in Southeast Asia which comprises of five sibling species namely A, B, C, D and E. However, only a few forms have been identified as malaria vectors in various endemic countries. Currently, for the first time egg morphometry and morphology has been used to differentiate the three known vector sibling species of Anopheles culicifacies collected from malaria endemic Madhya Pradesh state of central India. METHODS: The adult An. culicifacies (s.l.) was collected from five districts using standard mosquito collection methods. Adult female mosquitoes were allowed to lay eggs individually. The emerged mosquitoes were identified using allele specific polymerase chain reaction (AS-PCR) to sibling species. Eggs of sibling species A, D and E were studied using scanning electron microscopy (SEM) for morphometric and morphological characteristics. RESULTS: Currently AS-PCR identified four known sibling species (B, C, D and E) of An. culicifacies in the study area. The surface morphology and morphometric attributes of the sibling species A, D and E eggs considerably differed from each other. An. culicifacies E had a narrow deck as compared to A and D, while An. culicifacies A had a bigger micropyle with 6-7 sectors as compared to D and E that had 6 sectors. An. culicifacies D had the smallest float (the structure present on sides of the egg surface in which air is filled that help in floating) and the number of ribs was also fewer than for An. culicifacies A and E. CONCLUSIONS: The present study provides the first evidence that in addition to PCR assay, sibling species of An. culicifacies can also be differentiated using morphological and morphometric characteristics of the egg stage. The results also advocate that the sibling species of An. culicifacies are morphologically dissimilar and can be resolved using advanced microscopy.

Evaluation of Insecticides Susceptibility and Malaria Vector Potential of Anopheles annularis s.l. and Anopheles vagus in Assam, India.

During the recent past, development of DDT resistance and reduction to pyrethroid susceptibility among the malaria vectors has posed a serious challenge in many Southeast Asian countries including India. Current study presents the insecticide susceptibility and knock-down data of field collected Anopheles annularis sensu lato and An. vagus mosquito species from endemic areas of Assam in northeast India. Anopheles annularis s.l. and An. vagus adult females were collected from four randomly selected sentinel sites in Orang primary health centre (OPHC) and Balipara primary health centre (BPHC) areas, and used for testing susceptibility to DDT, malathion, deltamethrin and lambda-cyhalothrin. After insecticide susceptibility tests, mosquitoes were subjected to VectorTest™ assay kits to detect the presence of malaria sporozoite in the mosquitoes. An. annularis s.l. was completely susceptible to deltamethrin, lambda-cyhalothrin and malathion in both the study areas. An. vagus was highly susceptible to deltamethrin in both the areas, but exhibited reduced susceptibility to lambda-cyhalothrin in BPHC. Both the species were resistant to DDT and showed very high KDT50 and KDT99 values for DDT. Probit model used to calculate the KDT50 and KDT99 values did not display normal distribution of percent knock-down with time for malathion in both the mosquito species in OPHC (p<0.05) and An. vagus in BPHC (χ2 = 25.3; p = 0.0), and also for deltamethrin to An. vagus in BPHC area (χ2 = 15.4; p = 0.004). Minimum infection rate (MIR) of Plasmodium sporozoite for An. vagus was 0.56 in OPHC and 0.13 in BPHC, while for An. annularis MIR was found to be 0.22 in OPHC. Resistance management strategies should be identified to delay the expansion of resistance. Testing of field caught Anopheles vectors from different endemic areas for the presence of malaria sporozoite may be useful to ensure their role in malaria transmission.

Preliminary in vitro antiplasmodial activity of Aristolochia griffithii and Thalictrum foliolosum DC extracts against malaria parasite Plasmodium falciparum.

BACKGROUND: Resistance development in human malaria parasites against commonly used antimalarial drugs has necessitated the scientific exploration of traditionally used antimalarial plants. Plant derivatives have been used for curing malaria historically. The present study involves in vitro evaluation of two medicinally important plants Aristolochia griffithii and Thalictrum foliolosum DC used in antimalarial chemotherapy by the tribes of northeast India. METHOD: Chloroform, ethyl acetate and n-butanol extracts of Aristolochia griffithii and Thalictrum foliolosum DC were evaluated in vitro against chloroquine sensitive (SS) and chloroquine resistance strains (RS) of P. falciparum. The tests were conducted following WHO standard method and the inhibition of parasite (IC50) was calculated. RESULTS: In A. griffithii, the IC50 value for ethyl acetate extracts against SS was 6.2 ± 0.02 µg/ml and found to be lower than chloroform extracts, which exhibited an IC50 value of 14.1 ± 0.1 µg/ml (t = 191.1; p < 0.0001). The IC50 values of both chloroform and ethyl acetate extracts for RS were higher as compared to the SS (p < 0.0001). In T. foliolosum DC the IC50 concentration of chloroform extracts for SS and RS were 0.5 ± 0.0 and 1.1 ± 0.0 µg/ml respectively (t = 54.2; p < 0.0001). CONCLUSION: The present findings, although preliminary, but scientifically demonstrate that identification and isolation of active compounds of these two plant materials and testing against different Plasmodium species could lead to the development of potential antimalarial drugs for future.

Absence of asymptomatic malaria in a cohort of 133 individuals in a malaria endemic area of Assam, India.

BACKGROUND: Malaria in northeast India affects children and adults annually. The number of malaria cases reported has declined over the past few years. Nevertheless, it is not clear whether there is an actual decline in parasitaemia or whether asymptomatic malaria infections are on the rise, especially in forested and forest-fringed areas. Asymptomatic malaria forms a parasite reservoir that acts as an epicentre for malaria spread during high-transmission season. Therefore it is important to understand the quantum of asymptomatic malaria infections among the vulnerable population. METHOD: Four forest fringed historically malaria endemic villages were selected for the study. A total of 133 individuals without a fever history in the past four weeks were tested for malaria parasite using rapid diagnostic test (RDT), microscopy and polymerase chain reaction (PCR) assay during January - February 2014. Indoor resting Anopheles vectors were collected, identified and tested for sporozoite using VectorTest™ panel assay during October 2013 to March 2014, which is a low transmission season for malaria. Social and demographic data were recorded during the study. RESULTS: Mean age (± SEM) of the participants was 16.1 ± 1.2 years (95 % CI: 13.8-18.4). All participants (100 %) reported to use mosquito nets. Altogether, 43.6 % of participants had education below primary level and only 9 % reported a travel history during the past four weeks. All RDT, microscopy and PCR assays were found negative indicating no asymptomatic malaria parasitaemia. Seven known malaria vector species namely, Anopheles nivipes, An. minimus, An. annularis, An. vagus, An. aconitus, An. philippinensis and An. culicifacies, were recorded in the present study. VectorTest™ sporozoite panel assay conducted on 45 pools (N = 224) of vector mosquitoes were found negative for Plasmodium sporozoite. DISCUSSION: Northeastern states of India report asymptomatic malaria parasitemia along with high malaria transmission. An. minimus and An. dirus are recognised as efficient vectors, but An. culicifacies, An. philippinensis and An. annularis also play role in malaria transmission. Currently all participants were found negative for asymptomatic malaria, however the small sample size may restrict the scope of present results to the population living in more remote areas. CONCLUSION: No cases of asymptomatic malaria infections parasitaemia was found in the present study conducted during a low transmission season indicating that asymptomatic malaria parasitaemia may not be prevalent in the region. Mosquito specimens were tested negative for the malaria sporozoites. Study findings encourage the ongoing malaria intervention efforts and recommends similar investigations in different ecological areas involving large populations.

Essential oil based polymeric patch development and evaluating its repellent activity against mosquitoes.

Essential oil based insect repellents are environment friendly and provide dependable personal protection against the bites of mosquitoes and other blood-sucking insects. In the present study, optimized mixture of three essential oils was embedded into the ethylcellulose (EC) and polyvinylpyrrolidone (PVP K-30) polymers to develop essential oils based patch type mosquito repellent formulation. The developed formulation was characterized for various physico-chemical properties, oil release efficiency and essential oil-polymer interaction. Repellent activity of the formulation was evaluated against Ae. (S) albopictus mosquitoes and compared with commercially available synthetic insecticide based mosquito repellent cream Odomos(®) in the laboratory. The developed patches were 100% flat and there was no interaction between oil components and the excipients. Patches were smooth, homogenous and provided excellent mosquito repellent activity comparable to Odomos(®) under laboratory condition. Morphological and physico-chemical characterization indicated that the formulation was stable and suitable with the polymeric combination. The patch formulation did not show any inhalation toxicity in experimental Wistar rat. The repellent patches developed and evaluated currently, may provide a suitable, eco-friendly, acceptable and safe alternative to the existing synthetic repellent formulations for achieving protection against mosquitoes.

Multi-insecticide susceptibility evaluation of dengue vectors Stegomyia albopicta and St. aegypti in Assam, India.

BACKGROUND: Dengue is rapidly expanding mosquito-borne viral infection globally facing operational challenges due to insecticide resistance in dengue vectors. We have studied the susceptibility status of potential dengue vectors St. albopicta and St. aegypti to the commonly used insecticides. METHODS: Stegomyia larval bioassays were carried out to determine LC10, LC50 and LC99 values and resistance ratios (RR50 and RR99) for temephos. Adult susceptibility bioassay to 4% DDT, 0.05% deltamethrin, 5% malathion was assessed following standard procedure to determine the corrected mortality. Knock-down times (KDT50 and KDT99) were estimated and the knock-down resistance ratios (KRR50 and KRR99) were calculated. RESULTS: St. albopicta wild population (WP) of Sotia was resistant to temephos as the LC99 value was 0.12 mg/l and found to be 2.3 fold high than the reference population (RP). St. aegypti WP of Borgong, Kusumtola and Serajuli displayed a RR99 of 2.5, 5.4 and 4.5 respectively suggesting high level of resistance to temephos. Results suggested that both St. albopicta and St. aegypti WP were fully resistant to DDT (mortality < 90%) in all the study locations. Both the species were completely susceptible to deltamethrin and malathion (corrected mortality > 98%), except for St. albopicta at Sotia which displayed low level of resistance to malathion (corrected mortality =95.4%). The estimated KDT values for both the species indicated high level of knock-down resistance to DDT and susceptibility to deltamethrin. CONCLUSION: WP of both the dengue vectors showed varied response to temephos, while resistant to DDT and completely susceptible to deltamethrin. Both the species were susceptible to malathion at majority of the testing sites. Current results strongly advocate that DDT is no longer effective against dengue vectors, while thorough monitoring of malathion susceptibility in geographical area at dengue risk is inexorable to ascertain whether or not the resistance to malathion is focal. Information generated herein may be useful in better planning and implementing in dengue control strategy using insecticides.

Synergistic mosquito-repellent activity of Curcuma longa, Pogostemon heyneanus and Zanthoxylum limonella essential oils.

Mosquito repellents play an important role in preventing man-mosquito contact. In the present study, we evaluated the synergistic mosquito-repellent activity of Curcuma longa, Pogostemon heyneanus and Zanthoxylum limonella essential oils. The mosquito repellent efficacies of three essential oils were evaluated separately and in combination under laboratory and field conditions. N,N-Diethylphenylacetamide (DEPA) and dimethylphthalate (DMP) were used for comparison of the protection time of the mixture of essential oils. At an optimum concentration of 20%, the essential oils of C. longa, Z. limonella and P. heyneanus provided complete protection times (CPTs) of 96.2, 91.4 and 123.4 min, respectively, against Aedes albopictus mosquitoes in the laboratory. The 1:1:2 mixture of the essential oils provided 329.4 and 391.0 min of CPT in the laboratory and field trials, respectively. The percent increases in CPTs for the essential oil mixture were 30 for DMP and 55 for N,N-diethylphenylacetamide (DEPA). The synergistic repellent activity of the essential oils used in the present study might be useful for developing safer alternatives to synthetic repellents for personal protection against mosquitoes.