Behenic Acid as a multi-target inhibiting antibacterial phytochemical against Vibrio parahaemolyticus and Aeromonas hydrophila for effective management of aquaculture infections: an in-silico, in-vitro & in-vivo experimentation.

Multi-Target Inhibitors are the upcoming frontrunners of the antibiotic world as they provide significant advantage over drug resistance development. Antibacterial drug discovery research, requires more robust and innovative approaches such as multi-target inhibiting drugs, which over comes the innate hurdles in the field of antibiotics. In this current study, a curated set of 5,112 phytochemical molecules were virtually screened for its multi-target inhibition potential against 7 antibacterial protein drug-targets. Behenic Acid was identified to be the most significant phytochemical molecule with potential to inhibit Catalase Peroxidase (KatG), Adenylosuccinate Synthetase (ADSS) and Pyridoxine 5'-Phosphate Synthase (PdxJ), based on SeeSAR and AutoDock Vina results. Further, the inhibition potential of Behenic Acid was validated using 500 ns Molecular Dynamics (MD) Simulation based on Desmond analysis. Behenic Acid was further investigated in-vitro using agar-well-diffusion and Minimal Inhibitory Concentration (MIC) assay, where it demonstrated 20 ± 1mm zone-of-inhibition and 50 µg/ml MIC value against both Vibrio parahaemolyticus and Aeromonas hydrophila. Zebrafish based investigations was carried to confirm the in-vivo antibacterial efficacy of Behenic Acid. It was observed that, there is a progressive dose-dependent recovery from the bacterial infection, with highest recovery and survival observed in fishes fed with 100 µg/day of Behenic Acid. Results of the in-vitro and in-vivo assays strongly support the in-silico prediction of the antibacterial activity of Behenic Acid. Based on the results presented in this study, it is concluded that, Behenic Acid is a strong multi-target antibacterial phytochemical, that exerts antagonism against aquaculture bacterial pathogens such as V. parahaemolytics and A. hydrophila.Communicated by Ramaswamy H. Sarma.

Metagenomics reveals impact of geography and acute diarrheal disease on the Central Indian human gut microbiome.

BACKGROUND: The Central Indian gut microbiome remains grossly understudied. Herein, we sought to investigate the burden of antimicrobial resistance and diarrheal diseases, particularly Clostridioides difficile, in rural-agricultural and urban populations in Central India, where there is widespread unregulated antibiotic use. We utilized shotgun metagenomics to comprehensively characterize the bacterial and viral fractions of the gut microbiome and their encoded functions in 105 participants. RESULTS: We observed distinct rural-urban differences in bacterial and viral populations, with geography exhibiting a greater influence than diarrheal status. Clostridioides difficile disease was more commonly observed in urban subjects, and their microbiomes were enriched in metabolic pathways relating to the metabolism of industrial compounds and genes encoding resistance to 3rd generation cephalosporins and carbapenems. By linking phages present in the microbiome to their bacterial hosts through CRISPR spacers, phage variation could be directly related to shifts in bacterial populations, with the auxiliary metabolic potential of rural-associated phages enriched for carbon and amino acid energy metabolism. CONCLUSIONS: We report distinct differences in antimicrobial resistance gene profiles, enrichment of metabolic pathways and phage composition between rural and urban populations, as well as a higher burden of Clostridioides difficile disease in the urban population. Our results reveal that geography is the key driver of variation in urban and rural Indian microbiomes, with acute diarrheal disease, including C. difficile disease exerting a lesser impact. Future studies will be required to understand the potential role of dietary, cultural, and genetic factors in contributing to microbiome differences between rural and urban populations.

Recent trends in the impurity profile of pharmaceuticals.

Various regulatory authorities such as the International Conference on Harmonization (ICH), the United States Food and Drug administration (FDA), and the Canadian Drug and Health Agency (CDHA) are emphasizing on the purity requirements and the identification of impurities in Active Pharmaceutical Ingredients (APIs). The various sources of impurity in pharmaceutical products are - reagents, heavy metals, ligands, catalysts, other materials like filter aids, charcoal, and the like, degraded end products obtained during \ after manufacturing of bulk drugs from hydrolysis, photolytic cleavage, oxidative degradation, decarboxylation, enantiomeric impurity, and so on. The different pharmacopoeias such as the British Pharmacopoeia, United State Pharmacopoeia, and Indian Pharmacopoeia are slowly incorporating limits to allowable levels of impurities present in APIs or formulations. Various methods are used to isolate and characterize impurities in pharmaceuticals, such as, capillary electrophoresis, electron paramagnetic resonance, gas-liquid chromatography, gravimetric analysis, high performance liquid chromatography, solid-phase extraction methods, liquid-liquid extraction method, Ultraviolet Spectrometry, infrared spectroscopy, supercritical fluid extraction column chromatography, mass spectrometry, Nuclear magnetic resonance (NMR) spectroscopy, and RAMAN spectroscopy. Among all hyphenated techniques, the most exploited techniques for impurity profiling of drugs are Liquid Chromatography (LC)-Mass Spectroscopy (MS), LC-NMR, LC-NMR-MS, GC-MS, and LC-MS. This reveals the need and scope of impurity profiling of drugs in pharmaceutical research.