Introduction:Mass dog vaccination (MDV) is essential for eliminating dog-mediated rabies, responsible for over 95% of the estimated 74,000 annual human rabies deaths. Achieving ≥ 70% vaccination coverage necessary for herd immunity remains challenging, underscoring the need for effective vaccination strategies and reliable post-vaccination evaluation methods. Methods:MDV campaigns were conducted in four site in Bangladesh (two urban, two peri-urban) using three strategies: standard capture-vaccinate-release (CVR), enhanced CVR (eCVR), and roaming static point (RSP). Post-vaccination sight re-sight (SRS) and household surveys were used to characterize dog populations and estimate coverage. Three enumeration methods were compared to assess reliability and utility for campaign evaluation. Results:Over 12 working days, 9,195 dogs were vaccinated. eCVR achieved the highest operational efficiency (40.2 dogs/vaccinator/day), followed by standard CVR (36.6) and RSP (18.2). Post-vaccination surveys indicated that over 93% of dogs were free-roaming, and around 30% were unowned, highlighting limitations of static point strategies. Human-to-dog ratios (HDRs) were lower than the 100:1 planning estimate and varied widely across sites (mean: 67.8; range: 21.1-129.6), with no significant difference between urban and peri-urban areas (p = 0.479). Coverage estimates differed by enumeration method: 19% (dog density), 32% (HDR), and 47% (SRS), with comparable SRS- and HDR-based estimates (p = 0.920), and dog density formula estimates significantly lower (p = 0.014). Averaging across enumeration methods and sites, eCVR achieved the highest coverage (45%), followed by standard CVR (37%) and RSP (16%), with estimated RSP coverage significantly lower than eCVR (p = 0.028). Discussion:The wide heterogeneity in HDRs highlights the limitations of using a single ratio for national dog population extrapolation. Effective rabies control requires locally tailored vaccination strategies, real-time monitoring, and robust enumeration techniques to guide planning and ensure reliable evaluation of campaign impact.
The co-infection of DENV-1 and DENV-3 during endemic outbreaks can be potentially fatal and complicate the diagnostic process. In our study, we have focused on the development of a multiepitope vaccine against DENV-1 and DENV-3 co-infection, utilizing non-structural protein 1 (NS1) and envelope protein (E) as key antigens. B cell and T cell epitopes were predicted for their immunogenicity, antigenicity, and ability to elicit an IFN-γ response. The final construct showed predicted stability (Instability Index: 30.63), antigenicity (0.5509), non-allergenicity, and hydrophilic character (GRAVY: −0.226) based on computational assessments. Tertiary structural validation revealed 90.1% of residues in a favoured region. Molecular docking revealed a stronger binding of the DENV-TLR3 complex. The receptor and vaccine have stable interactions, according to molecular dynamics simulations and free energy estimations (-90 kJ/mol). Strong B and T cell memory responses were demonstrated by immune simulations, accompanied by increased levels of IgG, IFN-γ, and TGF-β. The codon-optimized sequence was successfully cloned into the pcDNA™3.1/V5-His-TOPO® expression vector for potential experimental validation. As a result of this in silico approach, a targeted vaccine for DENV-1 and DENV-3 co-infections is possible, which merits further experimental evaluation.
Introduction: Rapid urbanization in India has intensified the need for robust urban primary healthcare systems, especially for marginalized slum communities. This study presents a comprehensive, mixed-methods rapid assessment of Urban Ayushman Arogya Mandirs (USHC-AAMs) across 11 Indian states, analyzing their operational status, utilisation, service delivery, and governance in comparison to Urban Primary Health Centres (UPHCs). Methods: A mixed-methods rapid assessment, conducted from April to July 2025 in 11 states/UTs, combined quantitative analysis of service delivery and infrastructure data from national portals (HMIS, AAM, IPHS dashboards) with qualitative in-depth interviews of health department and urban local body officials, facility staff, and community beneficiaries. Data was analyzed for functionality, utilization, service delivery, human resources, governance, and coordination. Results: USHC-AAMs were found to be operational in most surveyed states, with rapid expansion in recent months. States like Kerala, Gujarat and Telangana demonstrated innovative upgrading of existing structures and robust financial/logistical support from urban local bodies, while others lagged due to infrastructure gaps and unclear administrative links. Results show that the USHC-AAM model has enabled decentralised, community-proximate health access, with improvements in non-communicable disease management and outpatient care footfall. However, persistent challenges remain, including infrastructure constraints, human resource shortfalls, inter-departmental coordination gaps, and funding sustainability. States with strong alignment between Health Departments and Urban Local Bodies, along with context-adapted implementation, reported better outcomes. Conclusion: The study recommends institutionalizing flexible planning per local need, dedicated funding streams, permanent infrastructure, sanctioned critical staffing, improved governance frameworks, and strategic partnerships, including public-private models for diagnostics and capacity building. These steps are essential for consolidating USHC-AAMs as the backbone of an equitable and resilient urban health architecture in India. Keywords: Urban primary healthcare, Ayushman Arogya Mandir, health system resilience, policy implementation
The bacterial endotoxin method uses amebocyte lysate reagents from the horseshoe crab (Limulus polyphemus or Tachypleus tridentatus) to detect and quantify bacterial endotoxin in injectable medications and medical devices. Two types of recombinant reagents have been introduced by various manufacturers, and the equivalency of their results to those of the natural Limulus amebocyte lysate (LAL) reagents has been recently assessed by the United States, European, and Japanese Pharmacopoeias. Recombinant reagents and amebocyte lysate reagents appear to be highly comparable, according to several investigations. LAL assays are currently the gold standard. LAL is used in vitro for the precise detection of endotoxins and is based on the endotoxin-activated Factor C-mediated clotting cascade. In our quality control laboratory, we tested several categories of drug samples using different compendial methods for detection of bacterial endotoxin that provide reliable and consistent results. Since 2024, a bacterial endotoxin test based on recombinant cascade reagent (rCR)-composed of the endotoxin sensor recombinant Factor C (rFC), Factor B, and pro-clotting enzyme inside of LAL-has been used as an animal-free alternative to LAL. The non-animal-derived reagent rCR uses cloned genes from the genome of the Limulus polyphemus horseshoe crab to detect and measure bacterial endotoxins. In addition to providing ethical and environmental benefits over traditional LAL assays, rCR eliminates interfering horseshoe crab blood components, allowing for extremely specific endotoxin detection. For rCR test performance in a routine setting, a comparative study was conducted of compendial bacterial endotoxin testing by LAL with the alternative animal-free reagent rCR, and the evidence was summarized and presented by analyzing different categories of pharmaceutical products. According to the compendial endotoxin testing requirements, all results were acceptable. When the rCR assay was applied rather than the LAL test, no interference was seen in certain samples. In this study, the rCR and LAL were equivalent and comparable.
Increasing amount of municipal solid waste is a growing global concern harming the health of the humans and animals and affecting the sustainability of the environment. While citizens are expected to take responsibility for waste sorting and segregation, limited engagement, lack of awareness, and dependency on others often hinder effective waste management at the source. This study aims to understand participants’ perceptions of waste segregation and management using a photovoice-based crowdsourcing approach. Through qualitative analysis of photographs and accompanying narratives submitted in a national contest, we explored how individuals interpret waste-related challenges, their motivations or barriers to action, and developed a conceptual framework. While many participants demonstrated awareness of the consequences of poor waste management and advocated for the principles of reduce, reuse, and recycle, a key theme that emerged was the lack of community participation and engagement. The conceptual framework was mapped against the Theory of Planned Behaviour and the Motivation-Opportunity-Ability-Behaviour model to contextualize and highlight the interaction of attitudes, perceived norms, and abilities. This study highlights the utility of participatory, photovoice-driven methods in shaping effective and inclusive waste management strategies.
