title: Anti-hemolytic, hemagglutination inhibition and bacterial membrane disruptive properties of selected herbal extracts attenuate virulence of Carbapenem Resistant Escherichia coli.
abstract: Expression of a multitude of virulence factors by multi-drug resistant microbial strains, e.g., Carbapenem Resistant Escherichia coli (Family: Enterobacteriaceae; Class: Gammaproteobacteria), is responsible for resistance against beta-lactam antibiotics. Hemolysin production and induction of hemagglutination by bacterial surface receptors inflicts direct cytotoxicity by destroying host phagocytic and epithelial cells. We have previously reported that Berberis aristata, Camellia sinensis, Cyperus rotundus Holarrhena antidysenterica and Andrographis paniculata are promising herbal leads for targeting Carbapenem resistant Escherichia coli. These herbal leads were analyzed for their anti-hemolytic potential by employing spectrophotometric assay of hemoglobin liberation. Anti-hemagglutination potential of the extracts was assessed by employing qualitative assay of visible RBC aggregate formation. Camellia sinensis (PTRC-31911-A) exhibited anti-hemolytic potential of 73.97 ± 0.03%, followed by Holarrhena antidysenterica (PTRC-8111-A) i.e., 68.32 ± 0.05%, Berberis aristata (PTRC-2111-A) i.e., 60.26 ± 0.05% and Cyperus rotundus (PTRC-31811-A) i.e., 53.76 ± 0.03%. Comprehensive, visual analysis of hemagglutination inhibition revealed that only Berberis aristata (PTRC-2111-A) and Camellia sinensis (PTRC-31911-A) exhibited anti-hemagglutination activity. However, Andrographis paniculata (PTRC-11611-A) exhibited none of the inhibitory activities. Furthermore, the pair wise correlation analysis of the tested activities with quantitative phytochemical descriptors revealed that an increased content of alkaloid; flavonoids; polyphenols, and decreased content of saponins supported both the activities. Additionally, flow cytometry revealed that cell membrane structures of CRE were damaged by extracts of Berberis aristata (PTRC-2111-A) and Camellia sinensis (PTRC-31911-A) at their respective Minimum Inhibitory Concentrations, thereby confirming noteworthy antibacterial potential of both these extracts targeting bacterial membrane; hemolysin and bacterial hemagglutination.
authors.affiliation: Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, India.
authors.forename: Pallavi
authors.initials: P
authors.lastname: Thakur
authors.affiliation: Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, India. Electronic address: ptrcmanuscripts@gmail.com.
authors.forename: Raman
authors.initials: R
authors.lastname: Chawla
authors.affiliation: Department of Biotechnology, Jamia Hamdard, Delhi, India.
authors.forename: Alka
authors.initials: A
authors.lastname: Narula
authors.affiliation: Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, India.
authors.forename: Rajeev
authors.initials: R
authors.lastname: Goel
authors.affiliation: Directorate General-Life Sciences, Defence Research and Development Organisation, DRDO Bhawan, Delhi, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Arora
authors.affiliation: Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, India.
authors.forename: Rakesh Kumar
authors.initials: RK
authors.lastname: Sharma
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology*; D015780:Carbapenems / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects*; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects* / Q000302:isolation & purification / Q000472:pathogenicity; D005434:Flow Cytometry; D006384:Hemagglutination / Q000187:drug effects; D006454:Hemoglobins / Q000032:analysis; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008826:Microbial Sensitivity Tests; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D013053:Spectrophotometry; D014774:Virulence / Q000187:drug effects; D018440:beta-Lactam Resistance
chemical_list: D000900:Anti-Bacterial Agents; D015780:Carbapenems; D006454:Hemoglobins; D010936:Plant Extracts
keywords: Carbapenem Resistant Escherichia coli; Hemagglutination; Hemolysis; Membrane disruption; Natural plant products