The ability of Klebsiella pneumoniae to form biofilm renders the pathogen recalcitrant to various antibiotics. The difficulty in managing K. pneumoniae related chronic infections is due to its biofilm-forming ability and associated virulence factors, necessitating the development of efficient strategies to control virulence factors. This study aimed at evaluating the inhibitory potential of selected phytochemical compounds on biofilm-associated virulence factors in K. pneumoniae, as well as authenticating their antibiofilm activity. Five phytochemical compounds (alpha-terpinene, camphene, fisetin, glycitein and phytol) were evaluated for their antibacterial and anti-biofilm-associated virulence factors such as exopolysaccharides, curli fibers, and hypermucoviscosity against carbapenem-resistant and extended-spectrum beta-lactamase-positive K. pneumoniae strains. The antibiofilm potential of these compounds was evaluated at initial cell attachment, microcolony formation and mature biofilm formation, then validated by in situ visualization using scanning electron microscopy (SEM). Exopolysaccharide surface topography was characterized using atomic force microscopy (AFM). The antibacterial activity of the compounds confirmed fisetin as the best anti-carbapenem-resistant K. pneumoniae, demonstrating a minimum inhibitory concentration (MIC) value of 0.0625 mg/mL. Phytol, glycitein and α-terpinene showed MIC values of 0.125 mg/mL for both strains. The assessment of the compounds for anti-virulence activity (exopolysaccharide reduction) revealed an up to 65.91% reduction in phytol and camphene. Atomic force microscopy detected marked differences between the topographies of untreated and treated (camphene and phytol) exopolysaccharides. Curli expression was inhibited at both 0.5 and 1.0 mg/mL by phytol, glycitein, fisetin and quercetin. The hypermucoviscosity was reduced by phytol, glycitein, and fisetin to the shortest mucoid string (1 mm) at 1 mg/mL. Phytol showed the highest antiadhesion activity against carbapenem-resistant and extended-spectrum beta-lactamase-positive K. pneumoniae (54.71% and 50.05%), respectively. Scanning electron microscopy correlated the in vitro findings, with phytol significantly altering the biofilm architecture. Phytol has antibiofilm and antivirulence potential against the highly virulent K. pneumoniae strains, revealing it as a potential lead compound for the management of K. pneumoniae-associated infections.

Bacterial quorum sensing (QS) system regulates pathogenesis, virulence, and biofilm formation, and together they contribute to nosocomial infections. Opportunistic pathogens, such as Pseudomonas aeruginosa, rely on QS for regulating virulence factors. Therefore, blocking the QS system may aid management of various infectious diseases caused by human pathogens. Plant secondary metabolites can thwart bacterial colonization and virulence. As such, this study was undertaken to evaluate three extracts from the medicinal plant, Melianthus comosus, from which phytochemical compounds were identified with potential to inhibit QS-dependent virulence factors in P. aeruginosa. Chemical profiling of the three extracts identified 1,2-benzene dicarboxylic acid, diethyl ester, neophytadiene and hexadecanoic acid as the common compounds. Validation of antibacterial activity confirmed the same MIC values of 0.78 mg/mL for aqueous, methanol and dichloromethane extracts while selected guanosine showed MIC 0.031 mg/mL. Molecular docking analysis showed anti-quorum sensing (AQS) potential of guanosine binding to CviR’ and 2UV0 proteins with varying docking scores of −5.969 and −8.376 kcal/mol, respectively. Guanosine inhibited biofilm cell attachment and biofilm development at 78.88% and 34.85%, respectively. Significant swimming and swarming motility restriction of P. aeruginosa were observed at the highest concentration of plant extracts and guanosine. Overall, guanosine revealed the best swarming motility restrictions. M. comosus extracts and guanosine have shown clear antibacterial effects and subsequent reduction of QS-dependent virulence activities against P.aeruginosa. Therefore, they could be ideal candidates in the search for antipathogenic drugs to combat P.aeruginosa infections.

The crisis of antibiotic resistance necessitates the search of phytochemicals as potential antibacterial, anti-quorum sensing and antibiofilm forming agents. For the present study, fifteen (15) selected medicinal plants were evaluated to inhibit the biological activities of multi-drug resistant (MDR) pathogenic bacteria (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis) associated with diabetic foot ulcer. Antibacterial activities revealed noteworthy minimum inhibitory concentration (MIC) values ≤1 mg/mL for thirteen (13) out of the sixty (60) plant extracts screened. The potent extracts included Euclea natalensis ethyl acetate (0.25 mg/mL), Aloe ferox methanol (0.5 mg/ml) and Warburgia salutaris aqueous (0.5 mg/mL) extracts. Chemical profiling of the active extracts using gas chromatography-mass spectrometry (GC-MS) identified neophytadiene, guanosine, squalene, cis megastigma-5,8-diene-4-one and sorbitol as prevalent compounds among the active extracts. Anti-quorum sensing activities of E. natalensis (ethyl acetate), A. ferox (methanol) and W. salutaris (aqueous) extracts ranged from 4.81 - 58.34% with E. natalensis (ethyl-acetate) showing the highest activity. Molecular docking against CviR protein showed selected compounds having high docking scores with sorbitol showing the highest score of -7.04 kcal/mol. Warburgia salutaris aqueous extract exhibited the highest biofilm inhibition (73%) against E. coli. Euclea natalensis, Aloe ferox and Warburgia salutaris compounds act as antagonist of N-acyl homoserine lactone (AHL) signaling, thus may serve as candidates in antipathogenic and antibiofilm phytomedicine development for MDR foot ulcer bacterial pathogens.

The food industry relies on citrus fruits for juice, canned fruit, and jam, creating significant waste from peels, seeds, and pomace. This waste contains valuable phytochemicals like carotenoids, essential oils, (poly)phenols, pectin, and vitamins, which can be used as nutraceuticals or key ingredients in functional foods for managing diabetes and obesity. Repurposing citrus peel waste offers an excellent opportunity to advance biorefineries and the bioeconomy. Compounds derived from citrus have attracted attention for their potential therapeutic effects on diabetes and obesity, and their effectiveness depends on various mechanisms. This review summarises citrus-derived phytochemicals that inhibit α-glucosidase and pancreatic lipase in vitro, highlighting their potential as anti-diabetic and anti-obesity compounds. We also discuss progress in using molecular docking screening against key drug targets linked to type II diabetes and obesity. This review explores novel citrus phytochemicals for the development of nutraceuticals and functional food ingredients with enhanced health benefits.

α‐Glucosidase inhibitors (AGIs) are compounds used to treat type 2 diabetes (T2D) by preventing the breakdown of dietary starch into monosaccharides, which reduces their absorption by the body and lowers blood glucose levels. AGIs often cause gastrointestinal issues such as diarrhea and flatulence due to excessive α‐amylase inhibition, leading to excess residual starch reaching the colon and being fermented by microbes. There is a need to prospect for novel AGIs that are effective and have fewer adverse effects. This study investigated the potential of citrus‐derived flavonoids as AGIs targeting amylolytic enzymes: α‐amylase and α‐glucosidase. Firstly, flavonoids were extracted from Citrus reticulata (tangerines) peels using an ultrasound‐assisted methanolic procedure, followed by C18 column‐purification and profiling with liquid chromatography‐mass spectrometry. Select citrus peel‐derived flavonoids, quercetin (−9.2 kcal/mol) and rutin (−10.8 kcal/mol), and the commercial AGI, acarbose (−8.7 kcal/mol), showed strong binding affinities against α‐glucosidase. Molecular dynamics simulations of the compounds were also assessed, revealing flexibility and stability in response to ligand interactions with the α‐glucosidase. The in silico data correlated positively with the results from the in vitro inhibition assays; acarbose (Ki = 0.14 mg/mL), quercetin (Ki = 0.12 mg/mL) and rutin (Ki = 0.19 mg/mL) recorded low inhibition constant values. The cytotoxicity profile of the selected compounds was also conducted on Caco‐2 cells, with flavonoids showing no significant cytotoxic effects. Flavonoids could be used as AGIs with minimal gastrointestinal impacts, reducing residual starch entering the colon and decreasing glucose uptake.

The emergence of multidrug‐resistant (MDR) Klebsiella pneumoniae remains a global health threat due to its alarming rates of becoming resistant to antibiotics. Therefore, identifying plant‐based treatment options to target this pathogen’s virulence factors is a priority. This study examined the antivirulence activities of twelve plant extracts obtained from three South African medicinal plants (Lippia javanica, Carpobrotus dimidiatus, and Helichrysum populifolium) against carbapenem‐resistant (CBR) and extended‐spectrum beta‐lactamase (ESBL) positive K. pneumoniae strains. The plant extracts (ethyl acetate, dichloromethane, methanol, and water) were validated for their inhibitory activities against bacterial growth and virulence factors such as biofilm formation, exopolysaccharide (EPS) production, curli expression, and hypermucoviscosity. The potent extract on K. pneumoniae biofilm was observed with a scanning electron microscope (SEM), while exopolysaccharide topography and surface parameters were observed using atomic force microscopy (AFM). Chemical profiling of the potent extract in vitro was analysed using liquid chromatography‐mass spectrometry (LC‐MS). Results revealed a noteworthy minimum inhibitory concentration (MIC) value for the C. dimidiatus dichloromethane extract at 0.78 mg/mL on CBR‐ K. pneumoniae. L. javanica (ethyl acetate) showed the highest cell attachment inhibition (67.25%) for CBR‐ K. pneumoniae. SEM correlated the in-vitro findings, evidenced by a significant alteration of the biofilm architecture. The highest EPS reduction of 34.18% was also noted for L. javanica (ethyl acetate) and correlated by noticeable changes observed using AFM. L. javanica (ethyl acetate) further reduced hypermucoviscosity to the least length mucoid string (1 mm‐2 mm) at 1.00 mg/mL on both strains. C. dimidiatus (aqueous) showed biofilm inhibition of 45.91% for the ESBL‐positive K. pneumoniae and inhibited curli expression at 0.50 mg/mL in both K. pneumoniae strains as observed for H. populifolium (aqueous) extract. Chemical profiling of L. javanica (ethyl acetate), C. dimidiatus (aqueous), and H. populifolium (aqueous) identified diterpene (10.29%), hydroxy‐dimethoxyflavone (10.24%), and 4,5‐dicaffeoylquinic acid (13.41%), respectively, as dominant compounds. Overall, the ethyl acetate extract of L. javanica revealed potent antivirulence properties against the studied MDR K. pneumoniae strains. Hence, it is a promising medicinal plant that can be investigated further to develop alternative therapy for managing K. pneumoniae‐associated infections.