Abirami Arasu , Nagaram Prabha , Durga Devi , Praveen Kumar Issac , Khaloud Mohammed Alarjani , Dunia A. Al Farraj , Reem A. Aljeidi , Dina S. Hussein , Magesh Mohan , Jehad Zuhair Tayyeb , Ajay Guru , Jesu Arockiaraj
J. Microbiol. 2023;61(11):993-1011. Published online December 4, 2023
Listeria monocytogenes is an important food-borne pathogen that causes listeriosis and has a high case fatality rate despite
its low incidence. Medicinal plants and their secondary metabolites have been identified as potential antibacterial substances,
serving as replacements for synthetic chemical compounds. The present studies emphasize two significant medicinal plants,
Allium cepa and Zingiber officinale, and their efficacy against L. monocytogenes. Firstly, a bacterial isolate was obtained
from milk and identified through morphology and biochemical reactions. The species of the isolate were further confirmed
through 16S rRNA analysis. Furthermore, polar solvents such as methanol and ethanol were used for the extraction of secondary
metabolites from A. cepa and Z. officinale. Crude phytochemical components were identified using phytochemical
tests, FTIR, and GC–MS. Moreover, the antibacterial activity of the crude extract and its various concentrations were tested
against L. monocytogenes. Among all, A. cepa in methanolic extracts showed significant inhibitory activity. Since, the A.
cepa for methanolic crude extract was used to perform autography to assess its bactericidal activity. Subsequently, molecular
docking was performed to determine the specific compound inhibition. The docking results revealed that four compounds
displayed strong binding affinity with the virulence factor Listeriolysin-O of L. monocytogenes. Based on the above results,
it can be concluded that the medicinal plant A. cepa has potential antibacterial effects against L. monocytogenes, particularly
targeting its virulence.
Fusarium solani has drawn phytopathogenic, biotechnological,
and medical interest. In humans, it is associated with
localized infections, such as onychomycosis and keratomycosis,
as well as invasive infections in immunocompromised
patients. One pathogenicity factor of filamentous fungi is biofilm
formation. There is still only scarce information about
the in vitro mechanism of the formation and composition of
F. solani biofilm. In this work, we describe the biofilm formed
by a clinical keratomycosis isolate in terms of its development,
composition and susceptibility to different antifungals and
ultraviolet light (UV) at different biofilm formation stages.
We found five biofilm formation stages using scanning electron
microscopy: adherence, germination, hyphal development,
maturation, and cell detachment. Using epifluorescence
microscopy with specific fluorochromes, it was elucidated
that the extracellular matrix consists of carbohydrates, proteins,
and extracellular DNA. Specific inhibitors for these
molecules showed significant biofilm reductions. The antifungal
susceptibility against natamycin, voriconazole, caspofungin,
and amphotericin B was evaluated by metabolic activity
and crystal violet assay, with the F. solani biofilm preformation
to 24 h increased in resistance to natamycin, voriconazole,
and caspofungin, while the biofilm preformation
to 48 h increased in resistance to amphotericin B. The preformed
biofilm at 24 h protected and reduced UV light
mortality. F. solani isolate could produce a highly structured
extra biofilm; its cellular matrix consists of carbohydrate polymers,
proteins, and eDNA. Biofilm confers antifungal resistance
and decreases its susceptibility to UV light. The fungal
biofilm functions as a survival strategy against antifungals
and environmental factors.
Cellular DNA in prokaryotes is organized in nucleic acid-protein self-assemblies referred to as the nucleoid. The physical forces responsible for its stability inside the poor solvent properties of the cytoplasm and their functional implications are not understood. Studies on the organisation and functioning of the cytosol of cells largely rely on experimental protocols performed in highly dilute solutions using biochemically purified molecules, which is not a reliable substitute for the situation existing in vivo. Our current research interest is focused on the characterization of biological and physical forces determining the compaction and phase separation of DNA in Escherichia coli cytoplasm. We have emphasized the effect of excluded volume in solutions with high macromolecular concentrations (macromolecular crowding) upon self-association patterns of reactions. The prokaryotic cytosol was simulated by addition of inert polymer polyethylene glycol (PEG) (average molecular weight 20000), as an agent which afterwards facilitates the self-association of macromolecules. Fluorescence microscopy was used for direct visualization of nucleoids in intact cells, after staining with DAPI (4',6-diamidino-2-phenylindole dihydrochloride). Addition of the crowding agent PEG 20,000, in increasing concentrations generated progressively enhanced nucleoid compaction, the effect being stronger in the presence of 0.2 M NaCl and 5 uM MgCl_2. Under these conditions, the nucleoids were compacted to volumes of around 2㎛^3 or comparable sizes with that of living cells.