Molecular detection and analysis of virulence genes in multi-drug resistant Escherichia coli from infected broilers.

Escherichia coli associated infections are major threats in poultry industry owing to severe economic losses each year. This study was conducted to identify E. coli isolates, to evaluate their antibiotic sensitivity and to find out their virulence patterns from infected broilers of Sylhet city in Bangladesh. Using polymerase chain reaction, a total 20 isolates were identified as E. coli from 11 chickens, exhibiting symptoms like colibacillosis and/or diarrhea. All isolates were positive for type-1 fimbrial adhesion (fimH), followed by putative avian hemolysin (hlyF) in 17 isolates; while none of the isolates was amplified with intimin (eaeA). Among 10 tested antibiotics, 100% of the isolates (n = 20) showed resistance to ampicillin, amoxicillin and tetra-cycline; but they were 100% sensitive to gentamicin. Organ specific correlations of antibiotic sensitivity were obtained among the isolates through principal component analysis (PCA) and Agglomerative Hierarchical Clustering (AHC). The 16S rRNA data of two multi-drug resistant isolates revealed closed clustering with clinical E. coli strains which could be indication of their zoonotic potential. In conclusion, the results depict higher prevalence of fimH and hlyF genes and drug resistance patterns of E. coli isolates from broilers in Sylhet city of Bangladesh.

Screening and druggability analysis of some plant metabolites against SARS-CoV-2: An integrative computational approach.

The sudden outbreak of novel coronavirus has caused a global concern due to its infection rate and mortality. Despite extensive research, there are still no specific drugs or vaccines to combat SARS-CoV-2 infection. Hence, this study was designed to evaluate some plant-based active compounds for drug candidacy against SARS-CoV-2 by using virtual screening methods and various computational analyses. A total of 27 plant metabolites were screened against SARS-CoV-2 main protease proteins (MPP), Nsp9 RNA binding protein, spike receptor binding domain, spike ecto-domain and HR2 domain using a molecular docking approach. Four metabolites, i.e., asiatic acid, avicularin, guajaverin, and withaferin showed maximum binding affinity with all key proteins in terms of lowest global binding energy. The crucial binding sites and drug surface hotspots were unravelled for each viral protein. The top candidates were further employed for ADME (absorption, distribution, metabolism, and excretion) analysis to investigate their drug profiles. Results suggest that none of the compounds render any undesirable consequences that could reduce their drug likeness properties. The analysis of toxicity pattern revealed no significant tumorigenic, mutagenic, irritating, or reproductive effects by the compounds. However, withaferin was comparatively toxic among the top four candidates with considerable cytotoxicity and immunotoxicity. Most of the target class by top drug candidates belonged to enzyme groups (e.g. oxidoreductases hydrolases, phosphatases). Moreover, results of drug similarity prediction revealed two approved structural analogs of Asiatic acid i.e. Hydrocortisone (DB00741) (previously used for SARS-CoV-1 and MERS) and Dinoprost-tromethamine (DB01160) from DrugBank. In addition, two other biologically active compounds, Mupirocin (DB00410) and Simvastatin (DB00641) could be an option for the treatment of viral infections. The study may pave the way to develop effective medications and preventive measure against SARS-CoV-2. Due to the encouraging results, we highly recommend further in vivo trials for the experimental validation of our findings.

Immunoinformatics approaches for designing a novel multi epitope peptide vaccine against human norovirus (Norwalk virus).

Norovirus is known as a major cause of several acute gastroenteritis (AGE) outbreaks each year. A study was conducted to develop a unique multi epitope subunit vaccine against human norovirus by adopting reverse vaccinology approach. The entire viral proteome of Norwalk virus was retrieved and allowed for further in silico study to predict highly antigenic epitopes through antigenicity, transmembrane topology screening, allergenicity assessment, toxicity analysis, population coverage analysis and molecular docking approach. Capsid protein VP1 and protein VP2 were identified as most antigenic viral proteins which generated a plethora of antigenic epitopes. Physicochemical properties and secondary structure of the designed vaccine were assessed to ensure its thermostability, hydrophilicity, theoretical PI and structural behavior. Molecular docking analysis of the refined vaccine with different MHCs and human immune TLR8 receptor demonstrated higher binding interaction as well. Complexed structure of the modeled vaccine and TLR8 showed minimal deformability at molecular level. The designed construct was reverse transcribed and adapted for E. coli strain K12 prior to insertion within pET28a(+) vector for its heterologous cloning and expression, and sequence of vaccine constructs showed no similarity with human proteins. However, the study could initiate in vitro and in vivo studies regarding effective vaccine development against human norovirus.