Multidrug-resistant Enteroaggregative Escherichia coli (EAEC) is gaining recognition as an important diarrheagenic pathogen with enhanced virulence characteristics. In this study, six typical EAEC isolates from clinical and animal sources were characterized phenotypically and genotypically. All isolates demonstrated the characteristic stacked-brick adherence pattern on HEp-2 cells and were positive for key EAEC-associated genes (aggR, cvd432, fimA, ecp, and irp2). Antimicrobial susceptibility testing identified three isolates as multidrug-resistant (MDR), with resistance observed against antibiotics such as, tetracycline, ampicillin, and ciprofloxacin. ESBL production was confirmed in MDR strains via double-disc synergy, nitrocefin, and PCR assays targeting blaTEM and blaCTXM9 genes. Under simulated gut stress conditions (bile salts, acidic pH, and oxidative stress), MDR strains exhibited enhanced survival and faster growth kinetics compared to non-MDR strains. Biofilm assays revealed stronger biofilm formation by MDR strains on diverse surfaces, with confocal microscopy confirming greater bio-volume and viability of MDR biofilms. In vivo survival assays using Galleria mellonella larvae showed significantly higher virulence of MDR strains, with increased mortality rates over 96 hours. The findings highlight the enhanced stress tolerance, biofilm-forming capacity, and pathogenicity of MDR-EAEC strains, underscoring their potential role in persistent infections and public health concerns.

Developing sustainable and edible active packaging materials is critical to mitigate microbial contamination and enhance food safety. This study developed a biodegradable and edible active film incorporating green-synthesized silver-zinc oxide nanocomposites (Ag/ZnO NCs) entrapping cinnamaldehyde (Ag/ZnO-N) in a food-grade alginate matrix for preservation of chilled chicken meat. The Ag/ZnO-N exhibited antibacterial activity against multi-drug-resistant (MDR) enteroaggregative Escherichia coli, Salmonella spp., and methicillin-resistant Staphylococcus aureus, with a minimum inhibitory concentration and minimum bactericidal concentration (MBC) of 7.80 and 62.50 µg mL-1, respectively. UV-vis and Fourier-transform infra-red spectroscopic analyses confirmed nanoparticle formation and cinnamaldehyde entrapment, while X-ray diffraction and scanning electron microscopy revealed polycrystalline morphology with reduced lattice crystallinity. The NCs exhibited minimal cytotoxicity to Vero cells (83.35% viability at 10-5 mg mL-1). Alginate (5%) films incorporating Ag/ZnO-N at MBC levels demonstrated enhanced surface roughness by atomic force microscopy, functional group integration, and potent antioxidant capacity (74.50 ± 0.14% ABTS˙+ and 8.38 ± 1.18% DPPH radical scavenging). The films were non-inhibitory to commensal microflora and exhibited significant antibacterial efficacy against MDR pathogens. In an ex vivo study on vacuum-packed chicken meat stored for 15 days under chilling conditions, the film significantly (P < 0.05) reduced aerobic plate, psychrotrophic, E. coli, and S. aureus counts, while Salmonella spp. were undetected. Lipid oxidation remained negligible and inductively coupled plasma mass spectrometry confirmed the absence of Ag+ and Zn2+ migration. These findings demonstrate that alginate-Ag/ZnO-N film offers a safe, edible, functional, and environmentally sustainable biomaterial platform for meat preservation, supporting circular bioeconomy-driven food systems.

Rising levels of pollution caused by multi-drug-resistant (MDR) pathogens, industrial dyes, and toxic heavy metals underscore the urgent need for efficient, eco-friendly, and sustainable remediation strategies. In this context, the present study synthesized biochar from poultry litter and assessed its antioxidant, antibacterial, and antibiofilm activities against MDR pathogens of public health relevance. The photocatalytic degradation of industrial dyes-methylene blue (MB), crystal violet (CV), and rhodamine B (RhB) was also evaluated under sunlight, LED, and UV irradiation, along with the adsorption of heavy metals (Cr, Pb, and Fe). Biochar synthesis was confirmed through UV-Vis and Fourier transform infrared spectroscopy analyses, while thermogravimetricdifferential thermal analysis, X-ray diffraction, Brunauer-Emmett-Teller and electron microscopy demonstrated its high thermal stability, distinct crystallinity, and predominantly mesoporous architecture. Elemental profiling revealed high carbon and oxygen content along with essential mineral constituents, reflecting strong carbonization and enhanced suitability for environmental applications. The biochar effectively inhibited biofilm formation in the tested MDR strains and showed dose-dependent free radical scavenging activity. Among the photocatalytic assays, sunlight provided superior dye degradation, with RhB degrading more slowly than MB and CV across all light sources. In heavy metal removal, the biochar exhibited higher adsorption capacity for Pb(II) and Cr(VI) compared to Fe. Overall, the findings demonstrate that poultry-litter-derived biochar is a promising multifunctional material with strong potential for environmental remediation and photocatalytic applications.

Phosphonates are chemically distinctive natural products characterized by a stable carbon-phosphorus (C-P) bond and are traditionally associated with a limited number of well-studied bacterial genera. We performed comparative genome mining of four publicly available Chelatococcus asaccharovorans genomes along with related Chelatococcus species to investigate their potential for phosphonate biosynthesis. Genome analysis using antiSMASH identified putative phosphonate biosynthetic gene clusters (BGCs) in two of the four C. asaccharovorans strains, whereas no phosphonate BGCs were detected in the remaining C. asaccharovorans strains and other analyzed Chelatococcus species. Both identified clusters contained phosphoenolpyruvate mutase (PepM), responsible for C-P bond formation in phosphonate biosynthesis. Functional annotation using BLASTp and conserved domain database analyses revealed a conserved set of phosphonate-associated genes, including aminotransferases, phosphocholine cytidylyltransferase family protein, regulatory proteins, and S-adenosyl-L-methionine-dependent methyltransferase. Although antiSMASH-predicted cluster architectures differed between the two strains, the core phosphonate biosynthetic gene content was largely conserved. Comparison with the MIBiG database revealed moderate similarity to previously characterized phosphonate pathways, including FR-900098, dehydrophos, fosfomycin, and dehydrofosmidomycin, indicating that the identified clusters share core phosphonate biosynthetic components but are not closely related to any currently characterized pathway. The restricted occurrence of PepM-containing phosphonate BGCs within the analyzed dataset indicates strain-specific phosphonate biosynthetic potential and suggests that phosphonate biosynthesis is not uniformly distributed across the genus Chelatococcus. Collectively, our findings expand the phylogenetic scope of phosphonate biosynthesis and identify C. asaccharovorans as a previously unrecognized reservoir of putative phosphonate biosynthetic diversity. This genome-guided investigation provides a foundation for future experimental validation and natural product discovery efforts targeting novel phosphonate compounds.

Environmental persistence of Bacillus anthracis spores sustains anthrax transmission, necessitating rapid and field-deployable detection tools. This study aimed to develop and quantitatively evaluate a LAMP-assisted CRISPR-Cas12a assay for sensitive and specific detection of B. anthracis spores in soil and meat meal matrices. Two B. anthracis-specific target genes plasmid-encoded lef and chromosomal SNP locus (CR5)- were selected. Target-specific LAMP primers and CRISPR crRNAs were designed using Primer Explorer, CRISPOR, and RNAfold platforms. The LAMP-CRISPR/Cas12a assay was optimised for reaction conditions and evaluated for analytical sensitivity and specificity using UV-inactivated spores and closely related Bacillus spp. Environmental applicability was assessed via spiking experiments in sterile soil and meat meal using the GABRI recovery method. Field validation was performed on 100 samples from anthrax-endemic regions of India, with performance compared against the WOAH-recommended lef gene-based real-time PCR. LAMP-CRISPR-Cas12a assay achieved detection limits of 10 spores/ml (lef) and 102 spores/ml (CR5), with no cross-reactivity against related species. In spiked matrices, spore recovery ranged from 50 to 75%, and detection sensitivity remained consistent. Field evaluation demonstrated a sensitivity of 91.70% and specificity of 100%, with near-perfect agreement (κ = 0.95) relative to real-time PCR. The assay delivered results within ∼70 min, including amplification and detection. The LAMP-assisted CRISPR-Cas12a platform provides a rapid, sensitive, and cost-effective approach for environmental detection of B. anthracis spores. The minimal equipment requirements and high diagnostic accuracy support its applicability for field-level surveillance and biosafety monitoring in resource-limited settings.

Early diagnosis of leptospirosis remains challenging because the low and transient leptospiraemia during the acute phase often results in bacterial DNA concentrations below the analytical detection limit of conventional molecular assays. This proof-of-concept study evaluated the feasibility of integrating aptamer-functionalised silver nanoparticles (AgNPs) as a pre-analytical enrichment step with loop-mediated isothermal amplification (LAMP) to improve the molecular detection of pathogenic Leptospira interrogans. Unlike previously reported aptamer-nanoparticle systems that primarily function as biosensors, the proposed approach employs aptamer-functionalised AgNPs as an upstream enrichment platform prior to nucleic acid amplification. Silver nanoparticles were synthesised by sodium borohydride reduction and characterised using UV-visible spectroscopy, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and transmission electron microscopy (TEM), while aptamer conjugation was supported by a characteristic surface plasmon resonance shift from 390 to 404 nm. Compared with unfunctionalised AgNPs and unenriched controls, aptamer-functionalised AgNP enrichment resulted in significantly greater DNA recovery (21.6% versus 7.3% at 103 CFU/mL; p = 0.018). Enrichment-assisted LAMP achieved an analytical limit of detection of 10 CFU/mL, representing a 100-fold improvement over conventional LAMP (103 CFU/mL), and demonstrated improved amplification performance in plasma samples containing low bacterial concentrations. Preliminary evaluation using 52 clinically suspected canine samples yielded a diagnostic sensitivity of 88.5% and specificity of 100%. Although additional mechanistic studies, comprehensive nanoparticle characterisation and larger multicentre clinical validation are required, these findings demonstrate the feasibility of aptamer-functionalised AgNP enrichment as a promising pre-analytical strategy for improving molecular detection of low-abundance Leptospira in resource-limited laboratory settings.

The environmental persistence of Bacillus anthracis spores continues to challenge rapid detection and surveillance of anthrax, necessitating simple, field-deployable analytical tools. This study aimed to develop a peptide-directed IgY-based latex agglutination test (LAT) as a rapid, cost-effective analytical method for the selective detection of B. anthracis spores. Four species-specific peptides derived from protective antigen (PA) and S-layer (EA1) proteins were identified through bioinformatic screening and used to generate polyclonal IgY antibodies. Latex beads (1.25%) were functionalized with optimized antibody concentrations to develop the LAT. Analytical performance was evaluated in terms of sensitivity, specificity, and cross-reactivity using inactivated spores of B. anthracis and related Bacillus spp., with validation by indirect ELISA. Applicability in complex matrices was assessed through spiking studies in soil and meat meal samples, followed by evaluation using 257 field samples from anthrax-endemic regions, benchmarked against WOAH-recommended pag gene PCR. The optimized LAT (200 µg ml-1 IgY) achieved a detection limit of 105 spores per ml, with high specificity for peptides PA-1 and EA-1 and minimal cross-reactivity. ELISA corroborated the specificity of peptide-derived IgY antibodies. In spiked matrices, recovery efficiency ranged from 50-75%, with a practical detection limit of 106 spores per g. Field validation demonstrated 72.7% sensitivity and 100% specificity (κ = 0.83) relative to PCR. This study presents a novel peptide-based immunoanalytical platform for on-site biosurveillance and resource-limited analytical settings that integrates specificity, rapid response, and low cost, offering a practical alternative to conventional methods for environmental monitoring of B. anthracis.

Newcastle disease virus (NDV) is a highly contagious avian pathogen requiring effective antiviral strategies. This study evaluated the in vitro antiviral efficacy of chitosan nanoparticles (CNPs) and curcumin-loaded chitosan nanoparticles (Cur-CNPs) against virulent NDV isolates from poultry in Kerala, India. Screening of 40 suspected flocks by reverse transcription polymerase chain reaction (RT-PCR) targeting the fusion (F) gene revealed a 12.5 % positivity rate. Sequencing confirmed virulent strains based on amino acid motifs at the F protein cleavage site. Two isolates, L2/MIB/PKD/23 and B3/MIB/PKD/23, were propagated in embryonated chicken eggs, and their virulence was confirmed through hemagglutination, hemagglutination inhibition, and mean death time assays. Virus adaptation to cell culture demonstrated higher replication efficiency in chicken embryo fibroblast cells. CNPs and Cur-CNPs were synthesised via ionic gelation and characterized by spectroscopic and microscopic techniques. Cytotoxicity assessment determined minimum non-cytotoxic concentrations of 187.5 μg/mL for CNPs and 1.47 μg/mL for Cur-CNPs. Antiviral activity, evaluated by MTT assay, demonstrated the highest protection with Cur-CNPs (0.75:1 ratio), achieving 44.18 % and 47.12 % protection for the two isolates. Viral titre reduction assays indicated a decrease of 3.00 log10 in TCID50 and a twofold reduction in hemagglutination titres. Quantitative real-time PCR confirmed significant viral load reductions (p < 0.001) with Cur-CNPs compared to CNPs. These findings indicate that Cur-CNPs exhibit strong antiviral activity against NDV and may serve as potential alternatives to conventional antiviral agents for Newcastle disease control.

This study envisaged the synthesis of a novel porous cobalt-based framework (Co-MOF) using 1,4-diazabicyclo[2.2.2]octane (DABCO) for the first of its kind as an organic ligand. The synthesis of Co-MOF-D evidenced by the color change from deep emerald-green to light parmesan was then confirmed by physicochemical characterization and further assessed for the in vitro cytotoxicity in HEK 293T, RAW 264.7 and Vero cell lines as well as antibacterial activity against multi-drug-resistant (MDR) bacteria of public health importance. UV-Vis spectrum exhibited distinguishable surface plasmonic resonance peak at 400 nm, while Fourier transform infrared spectra peaks demonstrated the functional groups presented in the Co-MOF-D framework. While X-ray diffraction analysis confirmed the crystalline nature of Co-MOF-D, electron microscopy revealed an aggregated polycrystalline morphology of Co-MOF-D with a mean size of 11.06 +/- 1.16 nm. In addition, Co-MOF-D demonstrated MIC and well as MBC values ranged from 250 to 1000 and >= 1000 mu M, respectively, against the test strains of MDR bacteria. Regardless of the cell lines tested, dose-dependent viability was observed, with values not <77.21 +/- 1.30 % for HEK293T, 68.09 +/- 2.07 % for RAW 264.7, and 67.39 +/- 1.75 % for Vero cell lines at the highest tested concentration (1 x 10(2) M). Overall, this study proposes promising biomedical applications of Co-MOF-D due to its tuneable porosity that would facilitate efficient drug delivery and controlled release to tackle the public health menace of drug resistance.

This study sought to evaluate the antibacterial potential of carvacrol and eugenol entrapped in green synthesized silver/zinc oxide (Ag/ZnO) nanocomposites (NCs) against multi-drug-resistant (MDR) isolates of enteroaggregative E. coli (EAEC), Salmonella enterica Typhimurium and S. Enteritidis as well as to assess their antioxidant and cytotoxic capacity. Initially, UV-Visible (UV-Vis) spectra verified the entrapment of Ag/ZnO NCs with different concentrations of eugenol (Ag/ZnO-E) and carvacrol (Ag/ZnO-R), while Fourier transform infrared spectroscopy (FTIR) spectra confirmed the functional groups on Ag/ZnO-E and Ag/ZnO-R. The X-ray diffraction (XRD) peak intensities were either attenuated (38.2 degrees) or nearly disappeared (44.3 degrees) after entrapment. Scanning electron microscopy (SEM) suggested an agglomerated irregularly polycrystalline structure of Ag/ZnO-E and Ag/ ZnO-R. Minimum inhibitory and bactericidal concentrations of 7.81 and 62.50 mu g/mL, respectively, were observed irrespective of tested bacterial strains, at all levels of eugenol and carvacrol incorporation. Besides, the concentration-dependent viability observed in Vero cell lines for Ag/ZnO-E and Ag/ZnO-R was determined to be 52.35 +/- 0.49 % and 46.32 +/- 2.28 %, even at the highest concentration tested (10-1 mg/mL). Both Ag/ZnO-E and Ag/ZnO-R exhibited significant dose-dependent free radical scavenging activity, though lower than ascorbic acid. Overall, this study indicated that Ag/ZnO-E and Ag/ZnO-R were effective antioxidant and antibacterial agents against MDR foodborne pathogens, suggesting their potential use in functionalized packaging to enhance the shelf life of food.

Escalating contamination of water bodies with toxic synthetic dyes and antibiotic-resistant bacteria underscores an urgent need for effective, eco-friendly remediation strategies. This study investigated antioxidant, anti-biofilm, and photocatalytic potential of silver-zinc oxide nanocomposites (Ag/ZnO NCs) synthesized from methanolic extract of Curcuma longa. UV-Vis spectroscopy, X-ray diffraction, and electron microscopy of Ag/ZnO NCs confirmed the synthesis. The Ag/ZnO NCs exhibited antibacterial activity against multidrug-resistant (MDR) pathogens, with minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of 31.25 and 62.50 µg/mL, respectively. These pathogens included enteroaggregative E. coli (EAEC), Salmonella Typhimurium, S. Enteritidis, and methicillin-resistant Staphylococcus aureus (MRSA). DPPH- and ABTS-based antioxidant assays revealed dose-dependent increase in free radical scavenging activity. Biofilm formation on polystyrene plates was significantly (P < 0.001) inhibited across all the MDR test strains, with higher inhibition observed at 48 h, and against MRSA. Photocatalytic activity was assessed by exposing the bacteria to Ag/ZnO NCs under LED light (460 nm). At MIC (1X and 1/2X), MDR-EAEC was eliminated within 30–120 min, and Salmonella spp. and MRSA were undetectable at 120 min. Sub-MIC concentrations (1/5X and 1/10X) increased clearance time to 180–240 min. The Ag/ZnO NCs demonstrated significant dye degradation efficiency (23–85

This study aimed to synthesize and characterize silver-based metal–organic frameworks (Ag-MOFs) using 1,4-diazabicyclo[2.2.2]octane (DABCO) as the organic ligand and to assess their antibacterial and cytotoxic properties.

Abstract Background Drug-resistant pathogens and industrial dye wastes have emerged as critical global public health concerns, posing significant risks to human and animal health, as well as to environmental sustainability. Green synthesized nano absorbents were found to be a viable strategy for treating drug-resistant pathogens and in wastewater. Hence, this study endeavored the synthesis of piperine-driven nano-zinc oxide (ZnONPs) and evaluated them for antibacterial, antibiofilm, and photocatalytic disinfection potential against multi-drug resistant (MDR) foodborne strains of non-typhoidal Salmonella (NTS). Besides, the dye degradation potential of ZnONPs when exposed to UV, sunlight, and LED lights and their antioxidant capacity were assessed. Results Initially, in silico analysis of piperine revealed drug-likeliness with minimal toxicity and strong interaction between piperine and OmpC motifs of Salmonella spp. UV spectroscopy of ZnONPs revealed a prominent absorption peak at 340 nm, while PXRD analysis confirmed the hexagonal wurtzite structure of ZnONPs by exhibiting peaks at 30°, 35.6°, 41.3°, 43.6°, 44.3°, 48°, 53°, 58°, and 59.2°, which corresponded to the lattice planes (102), (110), (103), (200), (112), (004), (104), (210), and (211). Additionally, the TEM images demonstrated predominantly spherical ZnONPs with hexagonal wurtzite crystalline SAED pattern. The minimum inhibitory concentration and minimum bactericidal concentration values (µg/mL) of the ZnONPs were found to be 62.50 and 125, respectively. The ZnONPs were observed to be safe with minimal hemolysis (less than 2%) in chicken RBCs, and no cytopathic effects were observed in the MTT assay using HEK cell lines. The NPs were found to be variably stable (high-end temperatures, proteases, cationic salts, and diverse pH), and were tested safe towards commensal gut lactobacilli. Additionally, in vitro time-kill kinetic assay indicated that the MDR-NTS strains were eliminated after co-incubating with ZnONPs for 6 h. The photocatalytic studies exhibited complete bacterial elimination under visible light at 4 h. Interestingly, the ZnONPs significantly inhibited the biofilm formation in the crystal violet staining assay by MDR-NTS strains (P < 0.001) at 24 and 48 h. Besides, a dose-dependent reducing power assay and 2,2′- azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) assay were exhibited. Moreover, ZnONPs significantly degraded methylene blue, crystal violet, and rhodamine-B under different light sources (sunlight, UV light, and LED). Conclusions This study revealed a sustainable one-pot method of synthesizing ZnONPs from piperine, which might be used as a viable antibacterial candidate with antioxidant, antibiofilm, and photocatalytic properties with eco-friendly implications and wastewater treatment.

This study aimed to encapsulate the Cecropin A (1-7)-Melittin (CAMA) peptide within chitosan nanoparticles (CS NPs) and evaluate its antimicrobial activity against multi-drug-resistant (MDR) strains of non-typhoidal Salmonella, employing both in vitro and in vivo assays. The CAMA-loaded CS NPs, synthesized using the ionic gelation technique, exhibited a particle size of 219.31 +/- 38.24 nm and a zeta potential of 8.2 +/- 0.2 mV, as measured by dynamic light scattering. The encapsulation efficiency was 75.45 +/- 2.5 %. Additionally, Fouriertransform infrared spectroscopy, scanning electron microscopy and transmission electron microscopy determined the functional groups, morphology and size of the CAMA-loaded CS NPs. The in vitro release kinetics revealed a pH-dependent release profile, with the highest cumulative release observed within 24 h at an alkaline pH (8.20), followed by physiological pH (7.40). Intracellular antimicrobial efficacy was assessed using HEp-2 cell lines, where CAMA-loaded CS NPs effectively cleared intracellular MDR Salmonella strains. Furthermore, CAMAloaded CS NPs were found to be safe for use with sheep erythrocytes, HEp-2 and RAW 264.7 cell lines, and beneficial gut lactobacilli, and were stable in the presence of proteolytic enzymes and simulated biological fluids. In vivo assays in Galleria mellonella larvae demonstrated an improved survival rate, reduced bacterial count, and minimal cytotoxicity, as confirmed by the lactate dehydrogenase assay. These results correlated with histopathological examination, suggesting that CAMA-loaded CS NPs could be a promising therapeutic candidate for combating intracellular Salmonella.

Food safety is a critical global priority, as foodborne diseases continue to pose significant public health challenges. Ensuring effective hazard management strategies. A key tool for managing microbial risks is microbial risk assessment (MRA), which involves hazard identification, hazard characterization, exposure assessment, and risk characterization. Various MRA approaches - qualitative, semiquantitative, and quantitative - are employed depending on available data and complexity. Predictive microbiology, utilizing mathematical models, plays a vital role in simulating microbial behavior under different food conditions, enhancing risk predictions. Advances in molecular technologies, such as multi-omics and metagenomics, alongside rapid detection methods, offer new insights into microbial behavior and improve detection accuracy. Risk ranking tools enable targeted interventions by comparing microbial hazards across different food-pathogen scenarios. Integrating these strategies will improve food safety, reduce the burden of foodborne illnesses, and enhance public health globally.

Coxiellaburnetii is an airborne bacterial zoonotic pathogen that causes Q fever/coxiellosis in humans and animals. Although dogs are suspected of transmitting Q fever to humans in past outbreaks, the prevalence of C. burnetii in the Indian dog population and risk factors for infection remain unknown. In this study, 452 dogs from pet clinics in three Indian states were screened for coxiellosis using molecular (Trans-PCR, Com 1-PCR) and serological (IFAT) tests. C. burnetii DNA was detected in 0.44% of blood samples using Trans-PCR, and pathogen-specific antibodies were found in 4.20% of sera using IFAT. Contact with stray dogs and ownership by farmers were identified as risk factors for canine coxiellosis. This study appears to be the first systematic assessment of coxiellosis and associated risk factors among dogs in India. A large-scale assessment of canine coxiellosis and its risk factors is warranted among pets and high-risk occupational groups in India.

Nanotechnology has become a sustainable strategy to combat drug resistance. As agro-waste management has become a concern, efficient management of produced waste has been an imposing global issue. This study evaluated antioxidant, as well as antibacterial and antibiofilm potential of zinc oxide nanoparticles (ZnO NPs) synthesized by hydrothermal approach using ethanolic extract of ‘Monsooned Malabar Robusta coffee’ husk against multi-drug-resistant (MDR) strains of enteroaggregative Escherichia coli, Salmonella Enteritidis, S. Typhimurium and methicillin-resistant Staphylococcus aureus. The fabrication of ZnO NPs was confirmed by spectroscopy, whereas thermogravimetric analysis-differential thermogravimetric analysis confirmed stability of ZnO NPs, while the X-ray diffraction pattern confirmed the wurtzite crystalline structure. The agglomerated nature of ZnO NPs with a nearly spherical shape was evident with scanning electron microscopy, whereas transmission electron microscopy revealed a poly-crystalline nature with a mean diameter of 26.33±3.778 nm. The microbroth dilution technique revealed a minimum inhibitory concentration (MIC) of 250 μg/mL and a minimum bactericidal concentration (MBC) of 500 μg/mL for the ZnO NPs. Furthermore, ZnO NPs exhibited significant antibiofilm activity against the MDR-test strains. Moreover, ZnO NPs were tested safe at MIC and MBC doses in chicken erythrocytes, and commensal gut microflora tested were not inhibited. Besides, a dose-dependent antioxidant property was exhibited by ZnO NPs. Additionally, in vitro time-kill kinetic assay of MDR-test strains treated with ZnO NPs revealed a complete bacterial clearance at 24 h. Overall, the synthesis of ZnO NPs from coffee husk demonstrated a simple, eco-friendly and valorization approach that could be devised as a potential delivery molecule.