The role of SARS-CoV-2 genomic surveillance and innovative analytical platforms for informing public health preparedness in Bengaluru, India.

A comprehensive SARS-CoV-2 genomic surveillance programme that integrates logistics, laboratory work, bioinformatics, analytics, and timely reporting was deployed through a public-private partnership in the city of Bengaluru, Karnataka in India. As a result, 12461 samples have been sequenced and reported to the Karnataka State public health officials as time-sensitive, decision support during the last one year and uploaded in global public databases in a timely manner. This programme has developed an analytics platform for studying SARS-CoV-2 sequences and their epidemiological context. Continuous sequencing effort enabled timely detection of emergence of Omicron variant in India and the subsequent spread of the same and its sub-lineages with more logistic growth (BA.10, BA.12 and BA.5) in Bengaluru. Our data also helped to provide timely information on variants to determine which of the Variants of Concern tracked globally, were observed in Bengaluru, ensuring targeted efforts and reducing unwarranted fear. This effort highlights the importance of, and the urgent need to, increase genomic surveillance to support the states with limited sequencing and bioinformatics capacity. We describe the development and deployment of this end-to-end solution for genomic surveillance of SARS-CoV-2 in the city of Bengaluru.

In Vitro and In Vivo Antibiofilm Potential of Eicosane Against Candida albicans.

Candida albicans is the most prevalent fungus in humans, producing infections ranging from mucosal to systemic. C. albicans colonizes mucosal surfaces asymptomatically as commensal, but, if the host environment is disrupted, or if the host immune system is compromised, C. albicans can multiply and infect almost all places in the host. The present study was aimed to identify a promising antibiofilm agent against Candida albicans biofilm. Through the molecular docking approach, it was identified that Eicosane was the top hit among the alkanes screened. Furthermore, in vitro analysis revealed that Eicosane at 100 µg/mL was able to inhibit 60% of C. albicans biofilm without inhibiting the growth. Moreover, light microscopic investigation unveiled the significant reduction in the adhesion and colonization of yeast cells to the matrix on Eicosane-treated samples. The CLSM images showing a reduction in biomass and thickness of C. albicans biofilm in the presence of Eicosane were validated using COMSTAT. The results were well corroborated with SEM micrograph in which a pellucid gap between the cells was observed and colonization was considerably reduced. Further from qPCR analysis, the genes responsible for biofilm formation and hyphal growth were found to be downregulated in the presence of Eicosane. Similarly, Eicosane at BIC was able to significantly inhibit the adhesion and colonization of yeast cells on the chorion of the zebrafish embryos. Moreover, the binding ability of Eicosane to ALS3 was revealed through docking and molecular dynamics (MD) simulation studies.

Evaluation of antibiofilm potential of four-domain α-amylase from Streptomyces griseus against exopolysaccharides (EPS) of bacterial pathogens using Danio rerio.

Biofilm formation is a major issue in healthcare settings as 75% of nosocomial infection arises due to biofilm residing bacteria. Exopolysaccharides (EPS), a key component of the biofilm matrix, contribute to the persistence of cells in a complex milieu and defends greatly from exogenous stress and demolition. It has been shown to be vital for biofilm scaffold and pathogenic features. The present study was aimed to investigate the effectiveness of four domain-containing α-amylase from Streptomyces griseus (SGAmy) in disrupting the EPS of multidrug-resistant bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. In vitro analysis of preformed biofilm unveiled the antibiofilm efficacy of SGAmy against MRSA (85%, p < 0.05) and P. aeruginosa (82%, p < 0.05). The total carbohydrate content in the EPS matrix of MRSA and P. aeruginosa was significantly reduced to 71.75% (p < 0.01) and 74.09% (p < 0.01), respectively. The findings inferred from in vitro analysis were further corroborated through in vivo studies using an experimental model organism, Danio rerio. Remarkably, the survival rate was extended to 88.8% (p < 0.05) and 74.2% (p < 0.05) in MRSA and P. aeruginosa infected fishes, respectively. An examination of gills, kidneys, and intestines of D. rerio organs depicted the reduced level of microbial colonization in SGAmy-treated cohorts and these findings were congruent with bacterial enumeration results.

Suppression of Thiol-Dependent Antioxidant System and Stress Response in Methicillin-Resistant Staphylococcus aureus by Docosanol: Explication Through Proteome Investigation.

The present study was aimed to investigate the effect of docosanol on the protein expression profile of methicillin-resistant Staphylococcus aureus (MRSA). Thus, two-dimensional gel electrophoresis coupled with MALDI-TOF MS technique was utilized to identify the differentially regulated proteins in the presence of docosanol. A total of 947 protein spots were identified from the intracellular proteome of both control and docosanol treated samples among which 40 spots were differentially regulated with a fold change greater than 1.0. Prominently, the thiol-dependent antioxidant system and stress response proteins are downregulated in MRSA, which are critical for survival during oxidative stress. In particular, docosanol downregulated the expression of Tpx, AhpC, BshC, BrxA, and YceI with a fold change of 1.4 (p = 0.02), 1.4 (p = 0.01), 1.6 (p = 0.002), 4.9 (p = 0.02), and 1.4 (p = 0.02), respectively. In addition, docosanol reduced the expression of proteins involved in purine metabolic pathways, biofilm growth cycle, and virulence factor production. Altogether, these findings suggest that docosanol could efficiently target the antioxidant pathway by reducing the expression of bacillithiol and stress-associated proteins.