Molecular surveillance of dengue virus in field-collected Aedes mosquitoes from Bhopal, central India: evidence of circulation of a new lineage of serotype 2.

Introduction: Dengue fever is hyperendemic in several Southeast and South Asian countries, including India, with all four serotypes (DENV 1-4) circulating at different periods and in different locations. Sustainable and improved virological and entomological surveillance is the only tool to prevent dengue and other vector-borne diseases. Objectives: The present study has been carried out to detect and characterize the circulating dengue virus (DENV) in field-collected Aedes mosquitoes in Bhopal, Central India. Methods: Aedes mosquitoes were collected from 29 localities within Bhopal city during October 2020 to September 2022. DENV infection was assessed in the individual head and thorax regions of Aedes mosquitoes using reverse transcriptase PCR. Positive samples were sequenced, and the circulating serotypes and genotypes were determined using phylogenetic analysis. Results: DENV RNA was detected in 7 Aedes aegypti and 1 Aedes albopictus, with infection rates of 0.59 and 0.14%, respectively. Phylogenetic analysis revealed all the isolates belonged to DENV serotype 2 and distinctly clustered with the non-Indian lineage (cosmopolitan genotype 4a), which was not recorded from the study area earlier. The time to most common recent ancestor (TMRCA) of these sequences was 7.4 years old, with the highest posterior density (HPD) of 3.5-12.2 years, indicating that this new lineage emerged during the year 2014. This is the first report on the DENV incrimination in both Ae. aegypti and Ae. albopictus mosquitoes collected from Bhopal, Central India. Conclusion: The observed emergence of the non-Indian lineage of DENV-2 in Bhopal, which again is a first report from the area, coincides with the gradual increase in DENV cases in Bhopal since 2014. This study emphasizes the importance of DENV surveillance and risk assessment in this strategically important part of the country to decipher its outbreak and severe disease-causing potential.

An assessment of remotely sensed environmental variables on Dengue epidemiology in Central India.

In recent decades, dengue has been expanding rapidly in the tropical cities. Even though environmental factors and landscape features profoundly impact dengue vector abundance and disease epidemiology, significant gaps exist in understanding the role of local environmental heterogeneity on dengue epidemiology in India. In this study, we assessed the role of remotely sensed climatic factors (rainfall, temperature and humidity) and landscape variables (land use pattern, vegetation and built up density) on dengue incidence (2012-2019) in Bhopal city, Central India. Dengue hotspots in the city were assessed through geographical information system based spatial statistics. Dengue incidence increased from 0.59 cases in 2012 to 9.11 cases in 2019 per 10,000 inhabitants, and wards located in Southern Bhopal were found to be dengue hotspots. Distributed lag non-linear model combined with quasi Poisson regression was used to assess the exposure-response association, relative risk (RR), and delayed effects of environmental factors on dengue incidence. The analysis revealed a non-linear relationship between meteorological variables and dengue cases. The model shows that the risk of dengue cases increases with increasing mean temperature, rainfall and absolute humidity. The highest RR of dengue cases (~2.0) was observed for absolute humidity ≥60 g/m3 with a 5-15 week lag. Rapid urbanization assessed by an increase in the built-up area (a 9.1% increase in 2020 compared to 2014) could also be a key factor driving dengue incidence in Bhopal city. The study sheds important insight into the synergistic effects of both the landscape and climatic factors on the transmission dynamics of dengue. Furthermore, the study provides key baseline information on the climatic variables that can be used in the micro-level dengue prediction models in Bhopal and other cities with similar climatic conditions.

Influence of Host Blood Meal Source on Gut Microbiota of Wild Caught Aedes aegypti, a Dominant Arboviral Disease Vector.

Blood feeding is an important behavior of Aedes aegypti, a dominant arboviral disease vector, as it can establish and transmit viruses to humans. Bacteria associated with the mosquito gut can modulate the biological characteristics and behavior of disease vectors. In this study, we characterized the gut microbiota composition of human-blood-fed (HF), non-human-blood-fed (NHF) and non-fed (NF) field-collected Ae. aegypti mosquitoes, using a 16S metagenomic approach, to assess any association of bacterial taxa with the blood-feeding behavior of Ae. aegypti. A significant difference in the microbiota composition between the HF and NF mosquito group was observed. A significant association was observed in the relative abundance of families Rhodobacteraceae, Neisseriaceae and Dermacoccaceae in the HF group in contrast to NF and NHF Ae. aegypti mosquitoes, respectively. At the class level, two classes (Rhodobacterales and Neisseriales) were found to be in higher abundance in the HF mosquitoes compared to a single class of bacteria (Caulobacterales) in the NF mosquitoes. These results show that human-blood feeding may change the gut microbiota in wild Ae. aegypti populations. More research is needed to determine how changes in the midgut bacterial communities in response to human-blood-feeding affect the vectorial capacity of Ae. aegypti.

Diagnosis of Human Leptospirosis: Comparison of Microscopic Agglutination Test with Recombinant LigA/B Antigen-Based In-House IgM Dot ELISA Dipstick Test and Latex Agglutination Test Using Bayesian Latent Class Model and MAT as Gold Standard.

Leptospirosis is a spirochaetal infection that possesses a broad host range affecting almost all mammals. In the present study, the microscopic agglutination test (MAT) was compared with recombinant LigA/B antigen-based point-of-care diagnostics such as the in-house IgM dot ELISA dipstick test (IgM- DEDT) and the latex agglutination test (LAT) for the serodiagnosis of human leptospirosis. The comparison of the MAT with these two point-of-care diagnostics was performed using the MAT as the gold standard test and using Bayesian latent class modelling (BLCM), which considers all diagnostic tests as imperfect. The N-terminal conserved region of the LigA/B protein spanning the first to fifth big tandem repeat domains (rLigA/BCon1-5) was employed as a serodiagnostic marker in both of the bedside assays. A total of 340 serum samples collected from humans involved in high risk occupations were screened using the MAT, IgM DEDT and LAT. During the early phase of leptospirosis, BLCM analysis showed that the IgM DEDT and LAT had similar sensitivities (99.6 (96.0-100)) and (99.5 (95.2-100)), respectively, while the single acute phase MAT had the lowest sensitivity (83.3 (72.8-91.3)). Both the IgM DEDT and the LAT may be superior to the single acute phase MAT in terms of sensitivity during the early phase of infection and may be suitable for the early diagnosis of leptospirosis. However, BLCM analysis revealed that the use of both acute and convalescent samples substantially increased the sensitivity of the final MAT (98.2% (93.0-99.8%)) as a test to diagnose human leptospirosis. Both the IgM DEDT and LAT can be employed as bedside spot tests in remote locations where the MAT is not easily accessible.

Seroprevalence and risk factors of brucellosis in livestock in the wildlife and livestock interface area of Similipal Biosphere Reserve, India.

BACKGROUND AND AIM: Brucellosis is an important zoonotic disease that affects fertility in farm animals. The risk factors of brucellosis have not been well studied. This study aimed to understand the seroprevalence and risk factors of brucellosis among livestock in Bangriposi block of Mayurbhanj district in Odisha, a region that borders Similipal wildlife reserve. MATERIALS AND METHODS: Rose Bengal plate test (RBPT) was carried out to estimate the seroprevalence of the livestock in this region. Bivariate analysis was carried out to analyze the association between the variables and brucellosis. Binary logistic regression was performed to assess the risk factors associated with brucellosis in the livestock. RESULTS: Based on RBPT, the seroprevalence of brucellosis among cattle and goats was estimated to be 1.1% and 11.2%, respectively. Binary logistic regression analysis indicates that study area, age, goats, animals with a history of abortion, and rearing practices were the major risk factors in this region. CONCLUSION: This is one of the first studies in India to shed light on risk factors of brucellosis, an important neglected disease that affects the health of animals and humans and nation's economy.

Magnetic field induced synthesis and self-assembly of super paramagnetic particles in a protein matrix.

Aqueous solution of a globular protein named bovine serum albumin was homogeneously mixed with ferrous and ferric ions and allowed to gel at ambient conditions. Gels were then oxidized using sodium hydroxide, in the presence of magnetic field of magnitude 0.13 T. The effect of magnetic field on the above biomimetic synthesis was a reduction in particle size and a directional assembly of synthesized super paramagnetic particles into a regular pattern in the protein film. The microstructural revelation was complimentary to Mossbauer results and magnetic measurement studies, i.e., an interesting variation in the magnetic behaviour of self-assembled super paramagnetic particles as a function of dc magnetic field induced ordering.