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ABSTRACT: Objectives: The use of antibiotics in human medicine and livestock production has contributed to the widespread occurrence of Antimicrobial Resistance (AMR). Recognizing the relevance of AMR to human and livestock health, it is important to assess the occurrence of genetic determinants of resistance in medical, veterinary, and public health settings in order to understand risks of transmission and treatment failure. Advances in next-generation sequencing technologies have had a significant impact on research in microbial genetics and microbiome analyses. The aim of the present study was to compare the Illumina MiSeq and Ion Torrent S5 Plus sequencing platforms for the analysis of AMR genes in a veterinary/public health setting. Methods: All samples were processed in parallel for the two sequencing technologies, subsequently following a common bioinformatics workflow to define the occurrence and abundance of AMR gene sequences. The Comprehensive Antibiotic Resistance Database (CARD), QIAGEN Microbial Insight - Antimicrobial Resistance, Antimicrobial resistance database, and Comprehensive Antibiotic Resistance Database developed by CLC bio (CARD-CLC) databases were compared for analysis, with the most genes identified using CARD. Results: Drawing on these results, we described an end-to-end workflow for the analysis of AMR genes a using advances in next-generation sequencing. No statistically significant differences were observed among any other genes except the tet-(40) gene between two sequencing platforms, which may be due to the short amplicon length. Conclusions: Irrespective of sequencing chemistry and platform used, comparative analysis of AMR genes and candidate host organism suggest that the Illumina MiSeq and Ion Torrent platforms performed almost equally. Regardless of sequencing platform, the results were closely comparable with minor differences.

Emerging SARS-CoV-2 variants with higher transmissibility and immune escape remain a persistent threat across the globe. This is evident from the recent outbreaks of the Delta (B.1.617.2) and Omicron variants. These variants have originated from different continents and spread across the globe. In this study, we explored the genomic and structural basis of these variants for their lineage defining mutations of the spike protein through computational analysis, protein modeling, and molecular dynamic (MD) simulations. We further experimentally validated the importance of these deletion mutants for their immune escape using a pseudovirus-based neutralization assay, and an antibody (4A8) that binds directly to the spike protein's NTD. Delta variant with the deletion and mutations in the NTD revealed a better rigidity and reduced flexibility as compared to the wild-type spike protein (Wuhan isolate). Furthermore, computational studies of 4A8 monoclonal antibody (mAb) revealed a reduced binding of Delta variant compared to the wild-type strain. Similarly, the MD simulation data and virus neutralization assays revealed that the Omicron also exhibits immune escape, as antigenic beta-sheets appear to be disrupted. The results of the present study demonstrate the higher possibility of immune escape and thereby achieved better fitness advantages by the Delta and Omicron variants, which warrants further demonstrations through experimental evidences. Our study, based on in-silico computational modelling, simulations, and pseudovirus-based neutralization assay, highlighted and identified the probable mechanism through which the Delta and Omicron variants are more pathogenically evolved with higher transmissibility as compared to the wild-type strain.

Cumin (Cuminum cyminum L.; Jeera) is a popular herbal seed spice used in culinary preparation in India. Gujarat and Rajasthan are the largest producer states of cumin seeds from India, while India is also the largest supplier and exporter of cumin across the globe. However, production of cumin is suffering from loss of crop production due to the plant pathogen infections, especially from Fusarium oxysporium sp. Rhizomicrobiome is the key modulator of plant health, revitalizing nutrients and disease response against plant pathogens. The secretion of different metabolites such as root exudates plays an important role in host plant rhizosphere microbial interactions influencing the plant health, growth and development, nutrient acquisition, and disease resistance. Therefore, in this research study, we have examined the microbial diversity from the healthy and fungal infected rhizosphere samples of the three different Gujarat Cumin (GC-2, GC-3, and GC-4) varieties using 16S ribosomal RNA (rRNA) gene sequencing on Ion Torrent S5 sequencing platform. The findings revealed the major dominant family represented by Bacillaceae, Solibacteraceae, Nostocaceae, Paenibacillaceae, Scytonemataceae, and Halothiobacillaceae, while at genera level of taxonomic abundance were represented by Bacillus, Candidatus Solibacter, Synechococcus, Nostoc, Anabaena, and Oscillatoria. The research findings should enhance our understanding of healthy and infected plant rhizosphere microbiome for better crop productivity, disease resistance and management of the crop varieties against plant pathogens.

Wastewater-based epidemiology (WBE) is a promising approach to understand the actual prevalence of COVID-19 disease at the community level. Different studies have cited the presence of SARS-CoV-2 in wastewater samples. In the present study, eighteen influent wastewater samples from different sewage treatment plants and pumping stations (5 samples from Vadodara city, 4 from Gandhinagar, and 9 from Ahmedabad city) were collected and analyzed for the presence of SARS-CoV-2 RNA in Gujarat state, India. The results showed the highest SARS-CoV-2 effective gene concentration in Vadodara (3078 copies/L), followed by Ahmedabad (2968 copies/L) and Gandhinagar (354 copies/L). On comparing the virus gene concentration in wastewater samples, the SARS-CoV-2 genetic material exhibited a positive relationship with the number of confirmed and active cases in in all three cities. However, a minor variation in SARS-CoV-2 effective gene concentration was seen between Vadodara and Ahmedabad despite a >2.5 and >1.5 folds differences in the cumulative number of confirmed and active cases, respectively. This may occur primarily due to the greater test positivity ratio in Vadodara (3.30%) than Ahmedabad (1.40%) and might be the higher number of asymptomatic patients in Vadodara. The study confirms the potential of the WBE that can be used at a large scale around the globe for better dealing with the pandemic situation.

Novel severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) has claimed more than 3.3 million lives worldwide and still counting. As per the GISAID database, the genomics of SARS-CoV-2 has been extensively studied, with more than 500 genome submissions per day. Out of several hotspot mutations within the SARS-CoV-2 genome, recent research has focused mainly on the missense variants. Moreover, significantly less attention has been accorded to delineate the role of the untranslated regions (UTRs) of the SARS-CoV-2 genome in the disease progression and etiology. One of the most frequent 5’ UTR variants in the SARS-CoV-2 genome is the C241T, with a global frequency of more than 95 %. In the present study, the effect of the C241T mutation has been studied with respect to the changes in RNA structure and its interaction with the host replication factors MADP1 Zinc finger CCHC-type and RNA-binding motif 1 (hnRNP1). The results obtained from molecular docking and molecular dynamics simulation indicated weaker interaction of C241T mutant stem-loops with the host transcription factor MADP1, indicating a reduced replication efficiency. The results are also correlated with increased recovery rates and decreased death rates of global SARS-CoV-2 cases.

This paper describes the additional data to our research article “Bacterial line of defense in Dirinaria lichen from two different ecosystems: First genomic insights of its mycobiont Diriniria sp. GBRC AP01” by Puvar et al. [1]. In this manuscript we are presenting the data obtained during the annotation of the genome enriched from metagenomic data from the lichen samples.