Blood typing has revolutionized the field of medical science since its discovery about a century ago. Besides its established role in life-saving blood transfusions, researchers have always been curious about the relationship between blood groups and human ailments. The effect of blood groups on disease outcomes, susceptibility, and mortality has been widely explored. According to a particular school of thought, the endemicity of diseases shapes the distribution of blood group frequency in human populations and exert selection pressure favoring one blood type over another. Here we discuss the scope and association of different blood groups in the context of malaria.

Background & objectives: Recently, the incidences of chikungunya, dengue and Zika infections have increased due to globalization and urbanization. It is vital that reliable detection tools become available to assess the viral prevalence within mosquito populations. Methods: Based on the previous publications on clinical diagnosis in human infections, for the first time, we described a customized triplex RT-qPCR protocol for simultaneous detection of chikungunya virus (CHIKV), dengue virus serotypes 1-4 (DENV1-4) and Zika virus (ZIKV) in mosquitoes. Results: In preliminary assessment to determine the specificity and sensitivity of primers and probes, all six targets were detected individually with the following thresholds as indicated by calculated pfu equivalents: 3.96x10⁰ for CHIKV, 3.80x10¹ for DENV1, 3.20x10¹ for DENV2, 8.00x10⁴ for DENV3, 1.58x10⁰ for DENV4, and 6.20x10⁰ for ZIKV When tested in a full combination of six targets (CDZ mix), CHIKV, DENV1-4 mix or ZIKV were all detected with the thresholds of 1.32x10⁰ for CHIKV, 3.79x10⁰ for DENV1-4 and 2.06x10⁰ for ZIKV All targets, individually or in full combination were detected in the mixtures of Aedes aegypti (L.) homogenate and viral lysates. A robust evaluation with three replicates in each of three plates for CHIKV, DENV1-4 and ZIKV individually or in full combination was conducted. In individual assays, CHIKV was detected to 3.96x10^-1, DENV1-4 to 1.14x10⁰ and ZIKV to 3.20x10⁰. In full combination assays, CHIKV was detected to 1.32x10⁴, DENV1-4 to 3.79x10¹ and ZIKV to 1.07x10⁰. Interpretation & conclusion: This triplex RT-qPCR assay appears to consistently detect all six targets and does not cross react with Ae. aegypti homogenate, making it a feasible, practical, and immediately adoptable protocol for use among vector control and other entities, particularly in the endemic areas of CHIKV, DENVs and ZIKV.

Background & objectives: A successful blood meal acquisition process by an adult female mosquito is accomplished through salivary glands, which releases a cocktail of proteins to counteract the vertebrate host’s immune homeostasis. Here, we characterize a salivary-specific Heme peroxidase family member HPX12, originally identified from Plasmodium vivax infected salivary RNAseq data of the mosquito Anopheles stephensi. Methods: To demonstrate we utilized a comprehensive in silico and functional genomics approach. Results: Our dsRNA-mediated silencing experiments demonstrate that salivary AsHPX12 may regulate pre-blood meal-associated behavioral properties such as probing time, probing propensity, and host attraction. Altered expression of the salivary secretory and antennal proteins expression may have accounted for salivary homeostasis disruption resulting in the unusual fast release of salivary cocktail proteins and delayed acquisition of blood meal in the AsHPX12 knockdown mosquitoes. We also observed a significant parallel transcriptional modulation in response to blood feeding and P. vivax infection. Interpretation & conclusion: With this work, we establish a possible functional correlation of AsHPX12 role in the maintenance of salivary physiological-homeostasis, and Plasmodium sporozoites survival/transmission, though the mechanism is yet to unravel.

Background & objectives: Mosquitoes are insects of public health importance that act as a vector to transmit various vector-borne diseases in humans including dengue, malaria, filariasis and yellow fever. The continually employed synthetic insecticides have developed resistance in mosquitoes. Nano-based botanical insecticides can be considered as the best alternative due to several advantages like being simple, non-pathogenic, biodegradable and safe to the environment. The present work reported the maximum larvicidal potential of green synthesized silver nanoparticles (AgNPs) derived from the leaf extract of Solanum xanthoearpum against the third instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus over its crude leaf extract. Methods: The synthesis of AgNPs was done by adding leaf extract into silver nitrate solution in a conical flask. The characterization of AgNPs was done using different techniques such as UV-Vis, SEM, TEM, XRD, DLS and SAED. FT-IR analysis was done to find out the compound responsible for bio-reduction of silver nitrate. Larvicidal activity of AgNPs was checked against An. stephensi, Ae. aegypti, and Cx. quinquefasciatus according to WHO standard protocol and toxicity was evaluated against Poecilia reticulate. Results: A change in colour was observed indicating the synthesis of AgNPs which was further confirmed by a strong surface plasmon resonance peak at 421nm under the UV-Vis spectrum. SEM and TEM micrographs exhibited that the most common shape of AgNPs was spherical. XRD spectrum showed crystalline nature of silver nanoparticles. FT-IR spectrum showed the presence of various functional groups such as carboxyl and hydroxyl which might be responsible for bio-reduction and capping of silver nanoparticles. Further, silver nanoparticles were very effective against An. stephensi, Ae. aegypti, and Cx. quinquefasciatus with LC₅₀ and LC₉₀ values of 1.90, 2.36, 2.93, 3.82, 4.31 and 7.63 ppm, respectively, as compared to aqueous leaf extract after 72 h of exposure and were non-toxic against non-target organism P. retieulata. Interpretation & eonelusion: From the above finding, it can be concluded that fabricated AgNPs can be promising eco-friendly tools for controlling mosquito vectors.

Background & objectives: Transmission of dengue virus by Aedes aegypti mosquito is one of the major global health concerns. The present study was aimed to explore the larvicidal potential of oil extracted from kinnow peel waste to be used as an efficient, economic and safe agent against Ae. aegypti. Methods: Kinnow peel oil was extracted and its five concentrations at 40, 50, 60, 70 and 80 ppm were tested against 4^th instar larvae of Ae. aegypti. Larval mortality (%) and LC₅₀ and LC₉₀ values of toxicity were determined followed by evaluation of the residual activity effect of its leftover effective concentration on larval mortality, development and emergence. Effect of storage (2, 4 and 6 months) on larvicidal potential of kinnow peel oil was also determined. Results: Out of the tested concentrations, 70 ppm of kinnow peel oil was found to be the effective concentration against 4^th instar larvae of Ae. aegypti. LC₅₀ and LC₉₀ toxicity values were 47.26 and 61.56 ppm, respectively. No residual activity effect in terms of larval mortality was found, however a significant delay in development (L4 to adult) was observed after placing new larvae in the leftover effective oil concentration. No effect of storage on larvicidal potential of 2, 4 and 6 months old kinnow peel oil in comparison to freshly extracted oil was observed. Interpretation & conclusion: Kinnow peel oil proved to have a good potential as a biolarvicide against Ae. aegypti and could be used as an effective and eco-friendly mosquito control agent in the future.

Background & objectives: The state of Himachal Pradesh is one of the hilly forested states of India. Warming of climate has been evidenced in the state due to the ongoing climate change which may cause the upsurge/introduction of mosquito-borne diseases. To curb disease transmission, an effective vector control strategy will be required. Methods: Insecticide susceptibility status of available malaria vectors was determined using the standard WHO method in six districts Kangra, Una, Mandi, Bilaspur, Solan and Mandi of the state. An. culicifacies and An. fluviatilis were tested against DDT (4%), malathion (5%) and deltamethrin (0.05%) using WHO insecticide susceptibility kits. Results: Overall, An. culicifacies was found resistant to DDT in all the six districts, susceptible to malathion in all districts except Bilaspur and Solan where it showed possible resistance. It was susceptible to deltamethrin in all the study districts. An. fluviatilis was resistant to DDT and susceptible to malathion and deltamethrin in Kangra and Una districts. Interpretation & conclusion: At present, indoor residual spraying (IRS) is not being undertaken in Himachal Pradesh. However, with the information generated through the present study, the state government can plan evidence-based IRS at least for focal spray in limited foci reporting malaria incidence.

Background & objectives: Crimean-Congo Hemorrhagic Fever (CCHF) is a deadly viral infection reported from more than 30 countries. It is considered a zoonosis↱ and tick bites are the main route of transmission in nature. So far, the virus has been identified in 31 species of hard (Ixodidae) and soft (Argasidae) ticks. The aim of this study was to determine the rate of CCHF virus infection in hard ticks from South-Khorasan province, east of Iran. Methods: In this study, 684 livestock including 302 sheep, 344 goats, 16 cows and 22 camels were sampled from Birjand, Qaen, Khusf, Darmian and Sarbisheh counties. Genus and species of the ticks were diagnosed under stereomicroscope according to valid morphological keys. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the CCHF virus genome based on S segment in 100 ticks. Results: RT-PCR detected CCHF virus genome in 7 out of 100 ticks. Positive ticks belonged to Hyalomma and Rhipicephalus genera. CCHF virus infected species were Rhipicephalus sanguineus, Hyalomma detritium and Hyalomma asiaticum. All the infected ticks were isolated from goat and sheep and were from Birjand county. Interpretation & conclusion: Our results suggest that Hyalomma and Rhipicephalus may be the main vectors of CCHF virus in the study area.

Background & objectives: Bacillus subtilis subsp. subtilis (VCRC B471) and Pseudomonas fluorescens (B426) produce mosquitocidal biosurfactant, surfactin and di-rhamnolipid. The objective of the study was to carry out a small-scale field evaluation of the two biosurfactants to determine the efficacy, application dosage, residual activity and frequency of application against Anopheles stephensi immatures in selected sites in Goa, India. Methods: Surfactin (VCRC B471) and di-rhamnolipid (VCRC B426) were formulated as aqueous suspensions (5% AS), and were applied at the dosages of 34, 51 and 68 mL/m² and 27, 41 and 54 mL/m² respectively. Two experiments were carried out with the two formulations. Results: Surfactin (VCRC B471) formulation was effective at all the dosages and there was sustained reduction (>80%) in immature density in the treated sites up to 18 days in experiment 1 and up to 15 days in experiment 2. No pupae were found in the treated sites throughout the study. Di-rhamnolipid (VCRC B426) formulation was also found to reduce the immature density in the treated sites up to 14 days in experiment 1 and up to 15 days in experiment 2. Interpretation & conclusion: For VCRC B471, the optimum application dosage determined was 51 mL/m² and for VCRC B426, 27mL/m². The formulations are to be applied fortnightly for effective control of Anopheles. The application dosage determined in the present study can be used for large scale field evaluation to assess their suitability for use in public health programmes for the control of Anopheles mosquitoes vectoring malaria.

Background & objectives: An. annularis van der Wulp (1884) is the secondary malaria vector of importance in India. In Jharkhand state it is present in almost all the districts abundantly and transmits malaria. The development of resistance to Dichlorodipheny ltrichloroethane (DDT) in An. annularis was reported from various parts of India. The main objective of this study was to generate information on insecticide susceptibility status of An. annularis to DDT, malathion, deltamethrin and permethrin in different districts of Jharkhand state. Methods; Adult An. annularis female mosquitoes were collected form villages of six tribal districts Simdega (Kurdeg and Simdega CHC), Khunti (Murhu and Khunti CHCs), Gumla (Bharno and Gumla CHCs), West Singhbhum (Chaibasa and Bada Jamda CHCs), Godda (Poraiyahat and Sunderpahari (CHCs) and Sahibganj (Borio and Rajmahal CHCs). Insecticide susceptibility status was determined by using WHO tube test method against prescribed discriminatory dosages of insecticides, DDT - 4.0%, malathion - 5.0%, deltamethrin - 0.05% and permethrin - 0.75%. Results: An. annularis was reported resistant to DDT in six districts, possible resistant to malathion in districts Gumla, Khuntiand Sahibganj and susceptible to deltamehrin (98% to100% mortality) and permethrin (100% mortality). Interpretation & conclusion: An. annularis, the secondary vector species is associated with the transmission of malaria reported resistant to DDT and susceptible to pyrerthroids deltamethrin and permethrin. In view of large-scale distribution of long-lasting insecticidal nets (LLINs) in all the districts, the response to synthetic pyrethroid needs to be periodically monitored to assess the effectiveness.

Background & objectives: Previously there were reports from all over India about the changing spectrum of severe manifestations of Plasmodium falciparum malaria. Consequently, the present retrospective study was conducted to compare the severity of malaria caused by P. falciparum and P. vivax during 2007–08 and 2017–18. Methods: The present study was conducted on the patients admitted with severe malaria in a classified malaria ward of a tertiary care hospital in Bikaner, Rajasthan (Northwest India) during 2007–08 and 2017–18. It included adult patients of both sexes belonging to all age groups. The diagnosis was done by peripheral blood film (PBF), rapid diagnostic test (RDT), and validated by polymerase chain reaction (PCR). All patients were treated with intravenous oral quinine. The specific individual malaria complications registered in 2007–08 and 2017–18 were treated by following the standard WHO protocol. Results: In 2007–08, severe manifestations caused by P. falciparum were dominated by thrombocytopenia (25.98%) followed by jaundice (24.39%), multi-organ dysfunction (MODS) (16.66%), severe anemia (16.17%), cerebral malaria (5.39%), bleeding manifestation (3.92%) and shock (0.49%). While in the same year, P. vivax associated clinical spectrum of complications were observed to be dominated by thrombocytopenia (26.47%) followed by jaundice (25.00%), MODS (14.70%), severe anemia (5.88%), cerebral malaria (5.88%), renal failure (4.41%), bleeding manifestation (2.45%), shock (0.98%) and acute respiratory distress syndrome (ARDS) (0.49%). However, in 2017–18, the clinical spectrum of malaria complications caused by both species has changed. Relative to P. falciparum infections, P. vivax individual complications like thrombocytopenia (51.78%) (p<0.001) followed by jaundice (19.13%) (p<0.001) and severe anemia (4.22%) (p<0.05) were found to have increased significantly. Interpretation & conclusion: Over the last decade there is an apparent spatial and temporal shift in the clinical manifestations of severe malaria caused by the both Plasmodium species. As evident from the patient’s data from 2007–08 and 2017–18, the severity is more inclined towards Plasmodium vivax than Plasmodium falciparum malaria. Moreover, individual P. falciparum-associated complications were decreased significantly in the Bikaner region of Rajasthan, India.

Background & objectives: This study focuses on modulating dexterity of some ecological variables of Aedes vittatus classically breeding in rocky habitats. The study provides a useful insight into ecological variables that underpin or hinder profuse breeding of Ae. vittatus in rock pools and its probable role in disease transmission. Methods: HANNA HI98129 pH/EC/TDS/TEMP meter was used in situ while standard protocols were used to determine other hydro-chemical variables. Aedes vittatus larvae were obtained with soup ladle and modified ladle dippers. D-frame net was used to capture macroinvertebrates while plankton net was used to obtain samples of microalgae. Tadpoles and water turtles were collected with fine mesh invertebrate net. Macrophytes were uprooted and identified at the Herbarium Unit, Department of Botany, Ahmadu Bello University, Zaria. The influence of physicochemical variables was correlated with distribution of Ae. vittatus using Principal Component Analysis. Regression and ANOVA were used to test for association between predictor variables and mosquito abundance and for the difference amongst inselbergs. Results: Linear larval density of Ae. vittatus in rock pools which tapered across Guinea savanna were obtained from twenty-one sites with average density of 139.6 in Sudan savanna. Guinea savanna had an average larval density of 75.5 with lower subsets of moving average densities compared to Sudan savanna. One hundred and sixty-one aquatic insects belonging to four insect orders cohabited rock pools with Ae. vittatus. Toads and frogs’ tadpoles were of Bufonidae and Pyxicephalidae families while water turtles belong to Emydidae. pH, TDS (ppm), EC (μs/cm) and alkalinity (mg/l) differed significantly (p<0.05) with the abundance of Ae. vittatus in rock pools. Temperature, depth, water hardness and total suspended solid had direct influence on the distribution of Ae. vittatus in rock pools across sites. Significant positive correlation exists between aquatic insects and abundance of Ae. vittatus. Hydroperiod length, concentration of nitrate and pH were determinants that leverage profuse breeding of Ae. vittatus and survival of rock pool biota. Interpretation & conclusion: Results revealed that the bearing influence of rock pool variables is inevitable for breeding of Ae. vittatus. A well defined measure of efficacy incorporating indigenous communities for sustained vector control on inselbergs will go a long way in decimating population of Ae. vittatus and limit the risk of spread of yellow fever hitherto areas not thriving.

Background & objectives: Malaria remains a public health problem in Côte d’Ivoire. To cope with this issue, the Ministry of Health established strategies through Long-Lasting Insecticidal Nets (LLINs) and artemisinin-based medicines. To better understand the influence of periodic mass distribution of LLINs on malaria transmission, this entomological survey was conducted in three regions of Côte d’Ivoire. Methods: Mosquitoes were sampled by Human Landing Catches (HLC) in urban and rural settings of Korhogo, Man and Abengourou. Mosquitoes were identified morphologically and by molecular methods. Plasmodium falciparum infection was assessed by ELISA, and the Entomological Inoculation Rates (EIR) were calculated for each species and setting. Results: Only An. gambiae s.l. was identified in Korhogo and in Abengourou while An. gambiae s.l. and An. funestus s.l. was reported in Man. An. coluzzii was the most abundant species of the An. gambiae siblings collected in Abengourou, and in Man while An. gambiae was most abundant in Korhogo. In urban settings, malaria vectors showed high aggressiveness (>11 bites per person per night) and the annual EIR was high (83.22-438.44 infectious bites per person per year). In rural settings, malaria vectors showed also high aggressiveness (19-52 b/p/n). The annual EIR is very high (>94 ib/p/yr). However, the weakest EIR was recorded in the northern region with 94.90 ib/p/yr. Interpretation & conclusion: This work indicates that malaria transmission remains high and heterogeneous across Côte d’Ivoire, despite repeated mass distribution of LLINs. Also, in Man, malaria transmission is more intense with the involvement of two main vectors. Furthermore, in the village of Korhogo, the EIR remained relatively low.

Background & objectives: Although, the number of considerable cases of dengue and lymphatic filariasis have been reported from Galle District, Sri Lanka in the past several years, contribution of abandoned boats to spread vector mosquitoes of Aedes and Culex in the coast is not well studied. Our aim was to assess the positive composition of different types of abandoned boats by larval vector mosquitoes to investigate their occurrence and habitat preference, and the monsoonal and co-existence variation of Aedes mosquitoes. Methods: The 4^th instar larvae of Aedes and Culex species from three different abandoned boat types in nine subsampling sites at three municipal areas in Galle District were collected during 2017–2019. In total, 15 sampling rounds were conducted in each site for three years duration as five sample rounds per year. Larval collections and identifications were carried out using standard techniques and identification keys according to WHO guidelines. The occurrence of mosquito larvae was analysed by boat type, locations within area and year. Further, the average infestation variation of Aedes mosquitoes were compared with monsoonal and co-existence changers. Results; Out of the total abandoned boats, majority (51%) were engine boats and, 32.7%, 16.35% were troller boats and canoe boats, respectively. Troller boats were highly infested boat type for vector mosquitoes. Aedes albopictus was the dominant vector in abandoned boats other than recorded Ae. aegypti and Culex quinquefasciatus. Culex quinquefasciatus showed comparatively low infestation percentages. The average infestation of Ae. albopictus showed a consistent pattern with monsoon variation. A competitive rapid invasion of Aedes aegypti was observed with the suppressing Ae. albopictus in abandoned boats after 2018. Interpretation & conclusion: Abandoned boats contribute noteworthy to spread of Aedes and Culex vector mosquitoes in coastal belt, Galle District. These study findings would be helpful for researchers and health authorities to design appropriate vector control measures and to mitigate future dengue and filariasis outbreaks.

Background & objectives: There is a need to evaluate novel techniques for dengue control in India. Several formulations of pyriproxyfen have been assessed for efficacy and duration of action. Pyriproxyfen is also used as a microencapsulated ready-to-use formulation against the Aedes vector. We evaluated a novel pyriproxyfen-based microencapsulated formulation. This slow-release, ready-to-use aqueous spray is a larvicidal formulation, and we assessed its efficacy and residual action through laboratory and semi-field trials against Aedes immature stages. Methods: The study was carried out as per the guidelines for laboratory and field/small-scale field testing of mosquito larvicides by the World Health Organization. The evaluation was conducted in laboratory and semi-field conditions from August to December 2018. We tested the novel formulation on three materials (plastic, ceramic, and enamel) in the laboratory for its action as an antilarval. Four containers of each kind were sprayed with the formulation and kept as replicates. Four controls were used in the laboratory trials - 120 larvae (third instar) were introduced in the replicates and the controls each. Readings were taken daily till complete adult emergence or larval and pupal mortality. In the semi-field trials, we applied this formulation to the inside of desert coolers and observed larvicidal and pupicidal activity over five months. Data is presented in numbers and percentages, along with mean and standard deviation. Adult emergence and Emergence Inhibition was calculated. Results: There was 100% adult emergence inhibition amongst the exposed larvae in the treated containers in the laboratory trials. In the untreated controls, adult emergence ranged from 80–95% in all types of containers. In the semifield trials, Inhibition Emergence was 100% in the treated desert coolers during the five months of the study period. Interpretation & conclusion: This advancement in insecticide formulation technology promises to make dengue control more effective and efficient.

Mediterranean spotted fever (MSF) is a tick-borne acute endemic infectious disease caused by Rickettsia conorii. While MSF may progress asymptomatically, it may lead to clinical pictures like severe hemorrhagic fever. In this article, we are presenting an MSF case with signs of high fever, headache, nausea, weakness and generalized maculopapular rash. The diagnosis of the female patient who had a history of contact with a tick-infested dog was confirmed with her clinical and laboratory data. The clinical and laboratory findings of the patient who was given doxycycline by 200 mg/day for 7 days were improved in a short time. Rickettsia conorii serology by indirect immunofluorescence assay method confirmed the diagnosis of MSF. In cases of severe sepsis accompanied by high fever and generalized maculopapular rash where the source of the infection cannot be determined in the short term, carefully questioning exposure to ticks by considering the existing geographical, seasonal and endemic environmental factors may be life-saving in terms of early diagnosis and treatment of MSF, which may become fatal even in the absence of eschars (tache noire). The symptomatology of hemorrhagic fever associated with Rickettsia conorii may be confused with that of sepsis in clinical practice.