Evidence on the prevalence of emerging and re-emerging tick- and flea-borne rickettsial agents in acute encephalitis syndrome endemic areas of northeast Uttar Pradesh, India.

Outbreaks of acute encephalitis syndrome (AES) with unknown aetiology are reported every year in Gorakhpur district, Uttar Pradesh, India, and Orientia tsutsugamushi, the rickettsial pathogen, responsible for scrub typhus has been attributed as the primary cause of AES problem. However, information on the prevalence of other rickettsial infections is lacking. Hence, this study was carried out to assess any occurrence of tick- and flea-borne rickettsial agents in villages reporting AES cases in this district. In total, 825 peridomestic small mammals were trapped, by setting 9254 Sherman traps in four villages with a trap success rate of 8.9%. The Asian house shrew, Suncus murinus, constituted the predominant animal species (56.2%) and contributed to the maximum number (87.37%) of ectoparasites. In total, 1552 ectoparasites comprising two species of ticks and one species each of flea and louse were retrieved from the trapped rodents/shrews. Rhipicephalus sanguineus, the brown dog tick, was the predominant species retrieved from the trapped rodents/shrews, and the overall infestation rate was 1.75 per animal. In total, 4428 ectoparasites comprising five tick species, three louse species and one flea species were collected from 1798 domestic animals screened. Rhipicephalus microplus was the predominant tick species collected from the domestic animals. The cat flea, Ctenocephalides felis, constituted 1.5% of the total ectoparasites. Of all the ectoparasite samples (5980) from domestic animals and rodents, tested as 1211 pools through real-time PCR assays, 64 pools were positive for 23S rRNA gene of rickettsial agents. The PCR-positive samples were subjected to multi-locus sequence typing (MLST). In BLAST and phylogenetic analysis, the ectoparasites were found to harbour Rickettsia asembonensis (n = 9), Rickettsia conorii (n = 3), Rickettsia massiliae (n = 29) and Candidatus Rickettsia senegalensis (n = 1). A total of 22 pools were detected to have multiple rickettsial agents. The prevalence of fleas and high abundance of tick vectors with natural infections of rickettsial agents indicates the risk of transmission of tick- and flea-borne rickettsial diseases in rural villages of Gorakhpur. Further epidemiological studies are required to confirm the transmission of these agents to humans.

Probable sociobehavioural and environmental risk factors for dengue transmission in Puducherry, India - An exploratory study.

Context: Dengue is one of the most extensively spread mosquito borne disease. Puducherry has experienced outbreaks during the post monsoon season almost every year since 2003. Understanding the dynamics of disease transmission and the conducive factors favourable for its spread is necessary to plan early control measures to prevent outbreaks. Objective: To describe the sociodemographic details of the dengue recovered cases, their clinical features, management, probable sociobehavioural and environmental risk factors for acquiring infection that could favour disease spread. Methodology: An exploratory descriptive study was conducted among 23 individuals recovered from dengue during the outbreak in Puducherry in 2018. An interview guide was used to elicit details regarding the course of illness from its onset until recovery as well as the probable sociobehavioural and environmental risk factors from each participant. Descriptive statistics were reported as frequency, percentage, and mean scores. Results: All 23 were primary cases of dengue with fever and myalgia being the commonest presentation. Two of them developed dengue haemorrhagic fever, of which one completely recovered. Five were found to have dengue-chikungunya coinfection. Lack of awareness about dengue, noncompliance regarding proper solid waste management and environmental sanitation among the public was clearly evident. Conclusion: Local transmission was evident as most cases did not have any relevant travel history outside the State and from the prevailing mosquitogenic environmental conditions. Dengue being a preventable disease can be controlled only with the active participation of all stakeholders including primary care physicians and the community.

VectorInfo: A web resource for medically important Indian arthropod disease vectors.

VectorInfo is a freely accessible web resource, emphasised on medically important Indian arthropods funded by Indian Council of Medical Research (ICMR) and maintained by one of its premier institute, Vector Control Research Centre (VCRC). VectorInfo elucidates and curates medically important Indian arthropod's biological, omics technologies to adopt a holistic view of the molecules that make up an organism, aimed at the detection of genomics, transcriptomics, proteomics, enzymes & pathways and immune specific genes. The nitty-gritty of VectorInfo is aimed at scrutinizing all the possible information on Indian disease vectors in a single window for the scientific community. The database affords 53 medically important Indian arthropod's biological and omics information well-structured and provided with downloadable facilities. In addition to this, huge number of research articles were mined in the quest for gathering the recommended insecticide targets and their mechanisms, that pave ways to design and develop novel lead molecules through computational means. This current up-to-date database contains 2,498 omics entries beneficial for the molecular studies and analysis. In order to maintain regular updates, user forms were provided for the scientific community to submit research data to the database administrator. The VectorInfo furthermore conveys various resources for vector control and diagnostics and the links to the crucial software tools used for the Bioinformatics analysis.

Evidence of natural infection of Orientia tsutsugamushi in vectors and animal hosts - Risk of scrub typhus transmission to humans in Puducherry, South India.

BACKGROUND: Scrub typhus infection is endemic in India and reported to be the major cause for acute encephalitis syndrome (AES) in humans. Periodic occurrence of scrub typhus cases and presence of pathogen in rodents were also reported in areas with human cases of scrub typhus in Puducherry. OBJECTIVES: This study was carried out to screen Orientia tsutsugamushi in rodent/shrew reservoirs and vectors in villages of Puducherry with no reports of human scrub typhus cases. METHODS: This study was conducted during October 2017 to January 2018 in ten randomly selected villages in Puducherry. Rodents/shrews in the peridomestic area were trapped using Sherman traps. Screening of O. tsutsugamushi in rodents/shrews and mite vectors was done by polymerase chain reaction (PCR). Weil-Felix test was done to screen antibodies against O. tsutsugamushi in rodent serum samples. RESULTS: Among the 54 rodents trapped, Suncus murinus was the major small animal and Leptotrombidium deliense was the major mite species retrieved. PCR screening revealed pathogen positivity in 8 rodent blood and 3 pooled mite samples. Phylogenetic analysis has shown that Kato was the circulating serotype of O. tsutsugamushi. None of the rodent serum samples was tested positive for antibodies against O. tsutsugamushi by Weil-Felix test. CONCLUSIONS: The presence of pathogen in both vectors and reservoir animal hosts imposes a risk for scrub typhus transmission to the inhabitants; hence, initiation of vector control measures before the start of winter is recommended in the study area. It is also recommended to screen scrub typhus in patients with undifferentiated acute febrile illness and AES.

Evaluation of long-lasting indoor residual spraying of deltamethrin 62.5 SC-PE against malaria vectors in India.

BACKGROUND: Deltamethrin 62.5 polymer-enhanced suspension concentrate (SC-PE) is one of the World Health Organization-approved insecticides for indoor residual spraying and was recommended to evaluate its residual activity for determination of appropriate spray cycles in different eco-epidemiologic settings. In the current study, efficacy of deltamethrin 62.5 SC-PE was evaluated against vectors of malaria and its impact on malaria incidence in a Plasmodium falciparum hyper-endemic area in Koraput district, Odisha State, India. METHODS: The trial had two comparable arms, arm 1 with residual spraying of deltamethrin 62.5 SC-PE and arm 2 with deltamethrin 2.5% WP (positive control). Comparative assessment of the impact of each intervention arm on entomological (density, parity, infection and human blood index), epidemiological (malaria incidence) parameters, residual efficacy and adverse effects were evaluated. RESULTS: Both the arms were comparable in terms of entomological and epidemiological parameters. While, deltamethrin 62.5 SC-PE was found to be effective for 150 days in mud and wood surfaces and 157 days in cement surfaces; deltamethrin 2.5% was effective only for 105 days on mud surfaces and 113 days on cement and wood surfaces. CONCLUSIONS: Deltamethrin 62.5 SC-PE had prolonged killing effectiveness up to 5 months. Hence, one round of IRS with deltamethrin 62.5 SC-PE would be sufficient to cover two existing malaria peak transmission seasons (July-August and October-November) in many parts of India.

Frequency and Clinical Significance of Localized Adverse Events following Mass Drug Administration for Lymphatic Filariasis in an Endemic Area in South India.

Fear of adverse events (AEs) negatively affects compliance to mass drug administration (MDA) for lymphatic filariasis (LF) elimination program. Systemic AEs are believed to occur because of killing of microfilariae, whereas localized soft tissue reactions might be due to the death of adult worms following therapy. Most AEs are mild and self-limited. However, localized AEs are sometimes more significant and of concern to participants. Here, we describe localized AEs that were noted during a large community study that evaluated the safety of a triple-drug regimen (ivermectin, diethylcarbamazine, and albendazole) for the treatment of LF in India. We have also discussed the importance of timely detection and careful management of AEs for preserving community confidence in MDA.

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

BACKGROUND: MAGNet 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. METHODS: The evaluation was carried out in six experimental huts located at Kandhaguda village in Malkangiri district, Odisha state following the WHO guidelines. RESULTS: The 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. CONCLUSIONS: MAGNet LN showed extended wash resistance retaining its bio-efficacy up to 25 washes and met the WHOPES requirement of passing Phase II evaluation.

Multiple insecticide resistance in Anopheles culicifacies s.l. (Diptera: Culicidae) in east-central India.

Development of resistance among the vectors to different insecticides poses a potential threat to vector control programme. Regular monitoring of susceptibility status of vector species to commonly used insecticides is recommended for planning appropriate vector control measures. In this communication, we have determined the phenotypic resistance of Anopheles culicifacies s.l., the major malaria vector against commonly used various insecticides in ten highly malaria endemic districts of Odisha State in east-central India. Bioassays were conducted before and after mass distribution of long-lasting insecticidal nets (LLINs) on field caught female mosquitoes with dichlorodiphenyl-trichloroethane (DDT), malathion and deltamethrin following the standard World Health Organization (WHO) guidelines. From the bioassays using 1x diagnostic concentrations (DC) recommended by WHO, we confirmed a higher frequency of resistant phenotype in An. culicifacies s.l. against DDT (range: 72% to 90%; average: 82%) compared to that against malathion(range: 17% to 34%; average: 26.7%) and deltamethrin (range: 14% to 24%; average: 19.1%) during 2018. Since, resistance to pyrethroid is detected; it is recommended to carry out bioassays of An. culicifacies s.l. exposing to higher concentrations of deltamethrin which would yield relevant information on the intensity of resistance and be useful to select suitable insecticide for resistance management vector control interventions.

Molecular modelling studies of kdr mutations in voltage gated sodium channel revealed significant conformational variations contributing to insecticide resistance.

Voltage gated sodium channels (VGSC) of mosquito vectors are the primary targets of dichlorodiphenyltrichloroethane (DDT) and other synthetic pyrethroids used in public health programmes. The knockdown resistant (kdr) mutations in VGSC are associated with the insecticide resistance especially in Anophelines. The present study is aimed to emphasize and demarcate the impact of three kdr-mutations such as L1014S, L1014F and L1014H on insecticide resistance. The membrane model of sodium transport domain of VGSC (STD-VGSC) was constructed using de novo approach based on domain and trans-membrane predictions. The comparative molecular modelling studies of wild type and mutant models of STD-VGSC revealed that L1014F mutant was observed to be near native to the wild type model in all the respects, but, L1014S and L1014H mutations showed drastic variations in the energy levels, root mean square fluctuations (RMSF) that resulted in conformational variations. The predicted binding sites also showed variable cavity volumes and RMSF in L1014S and L1014H mutants. Further, DDT also found be bound in near native manner to wild type in L1014F mutant and with variable orientation and affinities in L1014S and L1014H mutants. The variations and fluctuations observed in mutant structures explained that each mutation has its specific impact on the conformation of VGSC and its binding with DDT. The study provides new insights into the structure-function-correlations of mutant STD-VGSC structures and demonstrates the role and effects of kdr mutations on insecticide resistance in mosquito vectors.

Abundance & distribution of trombiculid mites & Orientia tsutsugamushi, the vectors & pathogen of scrub typhus in rodents & shrews collected from Puducherry & Tamil Nadu, India.

BACKGROUND & OBJECTIVES: Human cases of scrub typhus are reported every year from Puducherry and adjoining areas in southern India. However, information on the presence of causative agent, Orientia tsutsugamushi, and its vectors is lacking. Hence, the objective of the study was to find out the vector as well as pathogen distribution in rodents and shrews present in the scrub typhus-reported areas in southern India. METHODS: Trombiculid mites were collected by combing rats and shrews collected using Sherman traps and identified to species level following standard taxonomical keys. The serum samples of the animals were used for Weil-Felix test and the clots containing blood cells were used for DNA extraction and polymerase chain reaction (PCR). RESULTS: A total of 181 animals comprising four rodent species and one shrew species were collected from 12 villages. High proportion of chiggers was collected from the shrew, Suncus murinus (79.1%) and Rattus rattus (47.6%). A total of 10,491 trombiculid mites belonging to nine species were collected. Leptotrombidium deliense, the known vector of scrub typhus pathogen, was the predominant species (71.0%) and the chigger (L. deliense) index was 41.1 per animal. Of the 50 animals screened for the pathogen, 28 showed agglutination against OX-K in Weil-Felix test indicating the presence of antibodies against O. tsutsugamushi, the causative agent of scrub typhus. PCR carried out with the DNA extracted from blood samples of two of the animals were positive for GroEl gene of O. tsutsugamushi. INTERPRETATION & CONCLUSIONS: L. deliense index was well above the critical limit of chigger load, indicating that all the villages were receptive for high risk of transmission of scrub typhus to human. Pathogen positivity was higher among animals collected from villages recorded for higher chigger indices due to active transmission between the chigger mites and reservoir host animals. The results are suggestive of routine vector/pathogen surveillance at hot spots to initiate timely preventive measures.

Epidemiological assessment of eight rounds of mass drug administration for lymphatic filariasis in India: implications for monitoring and evaluation.

BACKGROUND: Monitoring and evaluation guidelines of the programme to eliminate lymphatic filariasis require impact assessments in at least one sentinel and one spot-check site in each implementation unit (IU). Transmission assessment surveys (TAS) that assess antigenaemia (Ag) in children in IUs that have completed at least five rounds of mass drug administration (MDA) each with >65% coverage and with microfilaria (Mf) levels <1% in the monitored sites form the basis for stopping the MDA. Despite its rigour, this multi-step process is likely to miss sites with transmission potential ('hotspots') and its statistical assumptions for sampling and threshold levels for decision-making have not been validated. We addressed these issues in a large-scale epidemiological study in two primary health centres in Thanjavur district, India, endemic for bancroftian filariasis that had undergone eight rounds of MDA. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence and intensity of Mf (per 60 µl blood) were 0.2% and 0.004 respectively in the survey that covered >70% of 50,363 population. The corresponding values for Ag were 2.3% and 17.3 Ag-units respectively. Ag-prevalence ranged from 0.7 to 0.9%, in children (2-10 years) and 2.7 to 3.0% in adults. Although the Mf-levels in the survey and the sentinel/spot check sites were <1% and Ag-level was <2% in children, we identified 7 "residual" (Mf-prevalence ≥ 1%, irrespective of Ag-status in children) and 17 "transmission" (at least one Ag-positive child born during the MDA period) hotspots. Antigenaemic persons were clustered both at household and site levels. We identified an Ag-prevalence of ~1% in children (equivalent to 0.4% community Mf-prevalence) as a possible threshold value for stopping MDA. CONCLUSIONS/SIGNIFICANCE: Existence of 'hotspots' and spatial clustering of infections in the study area indicate the need for developing good surveillance strategies for detecting 'hotspots', adopting evidence-based sampling strategies and evaluation unit size for TAS.

Description of Sergentomyia (Parrotomyia) rectangulata sp. nov. (Diptera: Psychodidae: Phlebotominae) from southern India.

Both sexes of Sergentomyia (Parrotomyia) rectangulata sp. nov. from southern India are described with illustrations. The specimens were collected in tree holes, termite mounds, cattle sheds and mud-walled, thatched roof human dwellings in rural areas. Our findings reveal similarity with Sergentomyia (Parrotomyia) modii Lewis.

Description of Sergentomyia (Parrotomyia) rectangulata sp. nov. (Diptera: Psychodidae: Phlebotominae) from southern India

Both sexes of Sergentomyia (Parrotomyia) rectangulata sp. nov. from southern India are described with illustrations. The specimens were collected in tree holes, termite mounds, cattle sheds and mud-walled, thatched roof human dwellings in rural areas. Our findings reveal similarity with Sergentomyia (Parrotomyia) modii Lewis.