Black fly (Simulium sp) composition, daytime biting activity and possible onchocerciasis infection in north-east, India.

Pupal stages of Simulium were collected and identified from various breeding habitats of twelve locations in northeastern India. Simulium flies, while attempting to land on human were collected between 07:00-17:00 hrs to understand the biting pattern. Seven species belonging to three sub-genera, Eusimulium (1), Gomphostilbia (1) and Simulium (5) belonging genus Simulium were encountered. Out of total seven species recorded, S. (E) aureohirtum, S. (G) tenuistylum and S. (S) rufibasis were predominant and shared 30.3%, 29.9% and 27.6% of total collection. Stream breeding habitat contributed 47.3% of total catch and was found to be preferred breeding habitat (p < 0.0001). S. (S) christophersi and S. (G) tenuistylum were recorded for the first time from the northeastern region of India. Simuliids biting rhythm showed bimodal pattern and were more active during sunny day (p < 0.0001). Microscopic dissection of simuliids (n = 266) did not incriminate simuliids as vector of onchocerciasis.

Nested PCR detection of Plasmodium malariae from microscopy confirmed P. falciparum samples in endemic area of NE India.

The present study evaluates the performance of OptiMAL-IT test and nested PCR assay in detection of malaria parasites. A total of 76 randomly selected blood samples collected from two malaria endemic areas were tested for malaria parasites using microscopy and OptiMAL-IT test in the field. PCR assays were performed in the laboratory using DNA extracted from blood spots of the same samples collected on the FTA classic cards. Of the total of 61 field confirmed malaria positive samples, only 58 (95%) were detected positive using microscopy in the laboratory. Sensitivity, specificity, positive predictive value, negative predictive value and false discovery rate of OptiMal-IT in comparison to the microscopy were 93%, 83%, 95%, 79% and 5%, respectively. On the other hand, the sensitivity and specificity of PCR assay were 97% and 100%, respectively, whereas positive predictive value, negative predictive value and false discovery rate were 100%, 90% and 0%, respectively. The overall performance of OptiMal-IT and PCR assays for malaria diagnosis was 76% and 97%, respectively. PCR assay enabled the identification of infection with Plasmodium malariae Laveran, 1881 in four samples misidentified by microscopy and Plasmodium-specific antigen (PAN) identified by the OptiMAL-IT test. In addition to the standard methods, such PCR assay could be useful to obtain the real incidence of each malaria parasite species for epidemiological perspectives.

Influence of water physicochemical characteristics on Simuliidae (Diptera) prevalence in some streams of Meghalaya, India.

BACKGROUND & OBJECTIVES: Simuliids (Diptera) are one of the medically important biting insects group and have worldwide distribution. Their immature stages proliferate in fast flowing river or stream water and have been considered as important ecological indicator. METHODS: Aquatic stages of simuliids were collected and speciated from 16 different fresh water rivers and streams. Water flow rate was determined and water samples were analysed for various water variables such as water temperature, pH, dissolved oxygen concentration (DOC), dissolved oxygen saturation (DOS), conductivity, total dissolved solute (TDS), turbidity, resistivity and salinity. Linear regression was used to determine relationship between simuliid density and water variables, whereas multiple regression was used to determine the fitness for the presence of simuliid species. Principal component analysis (PCA) was used to determine the water parameters association with simuliid distribution. RESULTS: Total 565 specimens comprising of three species namely, Simulium (S) barraudi Puri, S. (S) striatum Brunetti, and S. (S) himalayense Puri were recorded in the present study. Simulium barraudi was the most abundant (56.8%) and its density was high (χ2 = 289.3; df = 2; p <0.0001) as compared to the others. The average population size of each species was 188.3, whereas Simpson and Shannon-Wiener diversity indices were 0.4466 and 1.306 respectively. Linear regression showed that simuliid density was associated with the water flow rate. Principal component analysis indicated that the water parameters accounted for 42.25% variation along D1 axis, while 24.1% variation along D2 axis. Atleast two principal components have eigenvalue >1 and accounted for 32.6% of variation. INTERPRETATION & CONCLUSION: Our study provides new information on simuliid species association with breeding water parameters in a little studied region of high biological interest. Turbidity, water flow and pH are important water parameters affecting the simuliid species prevalence. Each simuliid species preferred different sets of physicochemical parameters of breeding habitat, which are specific to that particular species. Therefore, simuliid species community as a whole cannot be considered as a suitable indicator of the streams water quality. In addition to describing simuliids, the information provided herein will be useful for the conservation of aquatic ecology and environment in Meghalaya state of India.

Molecular detection and pathogenicity of a nucleopolyhedrovirus isolated from looper caterpillar (Hyposidra talaca), a tea pest.

Hyposidra talaca is a major defoliating pest of tea plants in north-eastern part of India. In this study, we look for variations (if any) in the attacking virus. Viral samples were collected from different regions of the northern part of West Bengal in India and were analyzed by PCR technique to study the variations across the region. The partial segment of the HytaNPV polyhedrin gene was cloned and sequenced. A 527 bp nucleotide sequence containing highly conserved region from polyhedrin gene of HytaNPV was observed. The blast homology search for studied polyhedrin gene showed 98% sequence identity with the sequence of previous reported NPV of H. talaca, H. infixaria and Buzura suppressaria. Pathogenicity study against second instar H. talaca indicated that the LC50 values ranged from 4.61 × 105 to 7.57 × 105 polyhedral occlusion bodies per ml (POBs/ml) and the LT50 values ranged from 4.2 to 6.66 days. Sequencing result reveals that the same HytaNPV strain dominates across this area and the pathogenicity indicates its potential as an alternative to chemical insecticides to control H. talaca.

Polymerase chain reaction detection of human host preference and Plasmodium parasite infections in field collected potential malaria vectors.

This study was carried out to determine the human host preference and presence of Plasmodium parasite in field collected Anopheles mosquitoes among four villages around a military cantonment located in malaria endemic Sonitpur district of Assam, India. Encountered malaria vector mosquitoes were identified and tested for host preference and Plasmodium presence using PCR method. Human host preference was detected using simple PCR, whereas vectorial status for Plasmodium parasite was confirmed using first round PCR with genus specific primers and thereafter nested PCR with three Plasmodium species specific primers. Out of 1874 blood fed vector mosquitoes collected, 187 (10%) were processed for PCR, which revealed that 40·6% had fed on human blood; 9·2% of human blood fed mosquito were harbouring Plasmodium parasites, 71·4% of which were confirmed to Plasmodium falciparum. In addition to An. minimus, An. annularis and An. culicifacies were also found positive for malaria parasites. The present study exhibits the human feeding tendency of Anopheles vectors highlighting their malaria parasite transmission potential. The present study may serve as a model for understanding the human host preference of malaria vectors and detection of malaria parasite inside the anopheline vector mosquitoes in order to update their vectorial status for estimating the possible role of these mosquitoes in malaria transmission. The study has used PCR method and suggests that PCR-based method should be used in this entire malarious region to correctly report the vectorial position of different malaria vectors.