BackgroundAccurate and rapid identification of dipteran vectors is integral for entomological surveys and is a vital component of control programs for mosquito-borne diseases. Conventionally, morphological features are used for mosquito identification, which suffer from biological and geographical variations and lack of standardization. We used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for protein profiling of mosquito species from North India with the aim of creating a MALDI-TOF MS database and evaluating it.MethodsMosquito larvae were collected from different rural and urban areas and reared to adult stages. The adult mosquitoes of four medically important genera, Anopheles, Aedes, Culex and Armigerus, were morphologically identified to the species level and confirmed by ITS2-specific PCR sequencing. The cephalothoraces of the adult specimens were subjected to MALDI-TOF analysis and the signature peak spectra were selected for creation of database, which was then evaluated to identify 60 blinded mosquito specimens.ResultsReproducible MALDI-TOF MS spectra spanning over 2–14 kDa m/z range were produced for nine mosquito species: Anopheles (An. stephensi, An. culicifacies and An. annularis); Aedes (Ae. aegypti and Ae. albopictus); Culex (Cx. quinquefasciatus, Cx. vishnui and Cx. tritaenorhynchus); and Armigerus (Ar. subalbatus). Genus- and species-specific peaks were identified to create the database and a score of > 1.8 was used to denote reliable identification. The average numbers of peaks obtained were 55–60 for Anopheles, 80–100 for Aedes, 30–60 for Culex and 45–50 peaks for Armigeres species. Of the 60 coded samples, 58 (96.67%) were correctly identified by MALDI-TOF MS with a score > 1.8, while there were two unreliable identifications (both Cx. quinquefasciatus with scores < 1.8).ConclusionsMALDI-TOF MS appears to be a pragmatic technique for accurate and rapid identification of mosquito species. The database needs to be expanded to include species from different geographical regions and also different life-cycle stages to fully harness the technique for entomological surveillance programs.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-2854-0) contains supplementary material, which is available to authorized users.

-Mosquito cytogenetic studies have revealed that the majority of disease vector species exist as cryptic species complexes. In relevance to this, the present results include the sequence analysis of r-DNA internal transcribed spacer 2 (ITS2) of two populations of Anopheles subpictus Grassi collected from Chandigarh (Pop.A) and Hoshiarpur, Punjab, India (Pop.B). These sequences were compared with fi ve other populations C, D, E, F and G worked out earlier. ITS2 sequence in Pop. A was 681 bp compared to 491 bp in Pop. B. All the sequences were G:C rich. In populations A-E, which form the Indian component of An. subpictus, the ts/tv frequencies ranged from 0.25 to 0.77. Populations F and G, which represent the Srilankan fauna, were much higher at 0.83 to 0.90 in inland and coastal populations respectively. The SRF revealed various common interspersed repeats in the form of dimers, trimers, tetramers, pentamers and polymers. The TG repeat motif, which was repeated 50 to 60 times, was common in Indian and Srilankan populations of An. subpictus. Further, there were as many as seven different types of polymers/repeats in populations A-E of Indian region but none was shared by F and G of Srilanka. Phylogeny analysis of ITS2 sequence had well supported India and Srilanka clades in accordance with their allopatric status.

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Vector control is an imperative method for the control of vector borne diseases. Over the last few decades, many methods have been developed for their control and the main goal of these strategies is to reduce the number of mosquito populations to overcome the epidemic situations. Though despite continuous efforts of the present interventions being deployed in the vector control programs we are unable to control the disease transmission and outbreaks. Therefore, it highlights the importance of exploring the challenges which are hindering the success of these strategies and also alternative solutions for the same so as to boost the vector control interventions.

Background & Objectives: Understanding the distribution and prevalence of three major mosquito borne diseases in an area is critical for the development of effective vector control strategies and public health interventions. The present study is therefore aimed to explore the epidemiological trend of malaria, dengue and chikungunya from 2012 to 2018 in Punjab. Methods: Quantitatively retrospective data was collected from Department of Health and Family Welfare, the National Vector Borne Disease Control Programme (NVBDCP), Punjab from 2012 to March 2018. The collected data was statistically analysed. Results: In case of malaria highest prevalent districts in Punjab are Mansa and Bathinda, for dengue Patiala, Ludhiana and S.A.S. Nagar and Patiala for chikungunya. Malaria was reported mainly from rural areas while dengue and chikungunya were found more in urban areas. For all three mosquito borne diseases, males were infected more as compared to females. Malaria cases were reported in months of August and September while dengue cases increased from July to November whereas chikungunya cases were highest in months of August to October. Conclusions: These findings help in concluding the trend analysis which in turn helps us to increase our focus on disease endemic districts along with boosting the vector control strategies in respective districts. Further the control of these mosquito borne diseases can be solved by employing adequate human resources, by increasing awareness in the community by conducting health camps and strengthening the entomological surveillance for timely reduction in the breeding of the vectors, especially before the repeated rise in cases during the period from July to November each year.

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Abstract:The present paper deals with the RAPD-PCR based genomic characterization of Culex quinquefasciatus Say which is a major vector of filariasis in several parts of the Indian subcontinent. One population of the test organism used in the study was procured from Goa (pop.A) while the other (pop.B) was collected from a village Nadasahib (20 kms from Chandigarh). The RAPD-PCR amplification of whole body homogenate of freshly hatched individual specimens was carried out by using three random primers: primer I-5'-GTCCCGACGA -3'; primer II-5' -TGATCCCTGG -3' and primer III-5'-GTGACGTAGG -3'. Primer I produced 5 distinct bands from the DNA of pop. A, whose base composition ranged from 200-1000 bp. Likewise, 7 bands ranging from 130-750 bp and 4 bands ranging from 270-950 bp were generated with primers II and III respectively. In case of pop.B, a total of 8 bands ranging from 200-1000 bp were generated with primer I. Similarly, a total of 6 bands ranging from 250-900 bp with primer II and 4 bands ranging from 180-950 bp with primer III were produced. Based on the band sharing coefficient and the application of Nearest Neighbour Joining (NJ) analysis it was found that primer I was more suitable for detecting genomic differences at the species and generic levels while primer II was ideal for detecting variations in the number of bp in RAPD generated bands among different populations of Cx. quinquefasciatus.