Listeriosis caused by Listeria monocytogenes is an important emerging foodborne disease worldwide and possesses remarkable environmental adaptability, enabling its evolution under diverse host and environmental conditions. To address this, the present study investigated the occurrence, virulence potential, biofilm-forming ability, and antimicrobial resistance profiles of L. monocytogenes isolated from animal- and plant-derived foods collected from four states of India during 2023–2025. A total of 941 food samples, comprising animal-derived foods (n = 690) and plant-derived foods (n = 251), were analyzed using EN ISO 11290-1:2017 protocols, VITEK® 2 identification, and duplex PCR confirmation. Overall, 21 isolates (2.23

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.

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.

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.

Developing rapid, accurate, and sensitive methods to detect bacterial pathogens such as Listeria monocytogenes and Salmonella typhimurium is very important, given the global rise in foodborne outbreaks. To address this, we developed a duplex real-time PCR assay with high-resolution melting analysis (qPCR-HRMA) to detect these pathogens in meat products. The assay was standardized and validated according to ISO 22118:2011. The assay was optimized for basic PCR parameters and melting rate for HRM analysis. The reaction sensitivity was determined to be 2 pg of DNA, equivalent to 124 copies for Listeria monocytogenes and 100 copies for Salmonella typhimurium. The method sensitivity was found to be 150 CFU/mL for both pathogens in spiked meat samples. The assay was validated with proficiency test samples and was finally used to test real-world samples, where 4 samples were detected positive for the pathogens. This assay holds significant potential for regulatory food testing and clinical investigations.

Emerging antimicrobial resistance (AMR) against common foodborne pathogens, has been observed in an increasing trend recently. The situation becomes grave when multi-drug resistance is reported toward the antibiotics of front-line therapy. Therefore, novel alternate therapeutics such as cationic antimicrobial peptides (AMPs), nanoparticles, etc., have attracted considerable attention to combat AMR. This study evaluated the in vivo antimicrobial efficacy of previously characterized bactenecin and green synthesized silver nanoparticles (AgNPs) against multidrug-resistant field isolates of Listeria monocytogenes and Staphylococcus aureus originating from poultry meat. To assess the in vivo antimicrobial efficacy, the Galleria mellonella (Wax moth) larval model is used as a substitute for traditional murine in vivo models which have ethical, financial, and logistical limitations. The micro broth dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of bactenecin and AgNPs against both pathogens. The AgNPs and bactenecin showed 64 and 128 mu M MBC against MDR-L. monocytogenes and MDR-S. aureus, respectively. Further, to assess the in vivo antimicrobial efficacy of these therapeutic candidates against both bacteria, an LD50 assessment, survival rate, bacterial count, melanization rate, hemocyte enumeration, and cytotoxicity assays were carried out in the G. mellonella larvae model. Overall, the infected G. mellonella larvae treated with either bactenecin or AgNPs revealed an improved survival rate, immunomodulatory effect, and reduced bacterial counts thus leading to a lesser degree of bacterial-induced cytotoxicity. However, the current study demonstrated that AgNPs was more efficacious in terms of MBC value, inducing hemocyte count and providing a steady decline in bacterial-induced cytotoxicity as compared to bactenecin, whereas, bactenecin exhibited a better survival rate in MDR-L. monocytogenes infected larvae. To conclude, the results suggest that both bactenecin and AgNPs as alternate therapeutic candidates showed an effective antimicrobial effect on MDR-L. monocytogenes and MDRS. aureus of poultry origin. However, further investigation of these therapeutic candidates against the targeted pathogens in appropriate animal models is recommended before its application in the target host.

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.

Coxiellosis in animals is caused by the zoonotic pathogen, Coxiella burnetii. Although the disease is of public health importance it remains underdiagnosed and underreported. The cross- sectional study was aimed to estimate the occurrence of the disease in livestock of study area and also to identify the risk factors associated with the disease in animals. Blood, serum, and vaginal swabs samples were collected from 200 ruminants (cattle, sheep, and goats), across various farms in Karnataka, India. These samples were then screened using ELISA and PCR (com1 and IS1111). A questionnaire was administered to the farm owners to collect the risk factor-related information. About 5.26% cattle, 12.3% sheep, and 12.5% goats were positive by ELISA. By PCR, 9.47% cattle, 9.3% sheep, and 10% goats were positive. Overall, the occurrence of 14.73%, 18.46% and 17.5% was estimated in cattle, sheep and goat, respectively. PCR targeting the IS1111 gene detected higher number of samples as positive as compared to the com1 gene PCR. Higher number of vaginal swab samples were detected as positive as compared to blood. History of reproductive disorders (OR: 4.30; 95%CI:1.95- 9.46), abortion (OR: 30.94; 95%CI:6.30- 151.84) and repeat breeding (OR:11.36; 95%CI:4.16- 30.99) were significantly associated with coxiellosis (p<0.005). Multivariable analysis by logistic regression model analysis suggested retained abortion, repeat breeding and rearing of animal in semi-intensive system as factors significantly associated with the infection. Cultural identification of the PCR positive samples were cultured using embryonated egg propagation and cell culture techniques and positivity was confirmed in six samples. Phylogenetic analysis of the com1 and IS1111 gene revealed clustering based on similar geographic locations. The study estimated the occurrence of the disease in the study area and identified the potential risk factors.

The study was carried out to investigate two important foodborne pathogens, Salmonella and Listeria spp. in five organic farms in Uttarakhand state in the year 2018. The samples from soil, manure, water, and plants/plant parts were collected and screened for pathogens. The covered isolates were assayed for their antimicrobial susceptibility and the presence of representative beta-lactamase antibiotic-resistant gene (ARG) by PCR. A total of 2.2% (11/500) of samples tested positive for the genus Salmonella. However, none of the samples tested positive for Listeria spp. All the Salmonella isolates were recovered from environmental sources. On serotyping, 4 isolates were identified as serogroup Group C1, 3 as Salmonella Miyazaki, 2 as Salmonella Virchow and 1 each as Salmonella Infantis and Salmonella Gabon. Two Salmonella isolates, belonging to S. Group C1 and one S. Miyazaki isolate were pan-susceptible to all the antibiotics tested, while S. Virchow and S. Infantis showed multidrug-resistant. On PCR screening for three β-lactamase resistant genes (blaCTX-M-9, blaTEM, and blaAmpC), only three S. Miyazaki isolates amplified the blaCTX-M-9 gene. The study highlights the presence of MDR Salmonella in the organic farm environment warranting further studies in this direction to ensure safe organic farm produce for consumers.

The present study successfully synthesized nano CoAl2O4 by the solution combustion method, employing sucrose as an ergonomic fuel. The synthesis of CoAl2O4 was confirmed by X-ray diffraction, Raman spectroscopy as well as electron microscopy, and a mean size of 70–80 nm was observed. Besides, the antioxidant, antimicrobial, photocatalytic and antibiofilm potentials of nano CoAl2O4 were evaluated against multi-drug resistant (MDR) bacterial strains. Initially, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CoAl2O4 (µg/mL) were determined to be within the ranges of 62.50–250 and 250–500, respectively. The poultry red blood cell-based hemolysis and secondary cell line-based MTT assays revealed concentration-dependent toxicity with an inhibitory concentration-50 (µg/mL) of 75.0 and 25.0 against MDA-MB468 and HT29 cell lines, respectively. Interestingly, CoAl2O4 was found to generate reactive oxygen species and revealed potential antioxidant properties by DPPH and reducing power assays. Moreover, the synthesized materials permeabilized the membranes of both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria and were found to be bactericidal and exhibited antibiofilm activity against the MDR- E. coli strain. The nano CoAl2O4, when treated with the MIC and 1/2X MIC, eliminated MDR-methicillin-resistant S. aureus within 60 and 120 min, respectively, and was found to be an excellent photosensitizer. In short, this study concludes that CoAl2O4 was found to be a promising antimicrobial candidate with antioxidant as well as antibiofilm properties for application in wastewater treatment and therapeutic purposes.

Bioinspired nanomaterials have widely been employed as suitable alternatives for controlling biofilm and pathogens due to their distinctive physico-chemical properties. This study explored the antibiofilm as well as photocatalytic potential of silver (Ag) nanoparticles (NPs) synthesized using the cell-free supernatant of Lactobacillus acidophilus for the disinfection of multi-drug-resistant (MDR) strains of enteroaggregative E. coli (EAEC), Salmonella Typhimurium, S. Enteritidis and methicillin-resistant Staphylococcus aureus (MRSA) on exposure to LED light. In addition, the removal of toxic cationic dyes i.e., methylene blue (MB), rhodamine B (RhB) and crystal violet (CV) was explored on exposure to sunlight, LED and UV lights. Initially, the synthesis of AgNPs was verified using UV- Vis spectroscopy, X-ray diffraction and transmission electron microscopy. The synthesized AgNPs exhibited MIC and MBC values of 7.80 and 15.625 µg/mL, respectively. The AgNPs exhibited significant inhibition (P < 0.001) in the biofilm-forming ability of all the tested MDR isolates. On exposure to LED light, the AgNPs could effectively eliminate all the tested MDR isolates in a dose-dependent manner. While performing photocatalytic assays, the degradation of RhB was observed to be quite slower than MB and CV irrespective of the tested light sources. Moreover, the sunlight as well as UV light exhibited better photodegradation capacity than LED light. Notwithstanding the light sources, RhB followed zero-order kinetics; however, MB and CV followed primarily second-order kinetics. The green synthesized AgNPs were found to be an effective photocatalytic as well as antifouling candidate that could be applied in therapeutics and wastewater treatment.

Coxiella burnetii infection is an emerging/re-emerging public health problem affecting several countries worldwide. In India, the disease is mainly underdiagnosed, creating hindrances in its effective control. This study investigated the occurrence of C. burnetii among apparently healthy cattle and cattle with a history of reproductive disorders by both PCR and indirect-ELISA. A total of 731 clinical samples (serum: 531, and vaginal swabs as well as blood: 100 each) from 531 cattle were screened for coxiellosis. The serum, blood, and vaginal swabs each collected from 100 cattle with a history of reproductive disorders were screened using Com1-PCR, Trans-PCR, and indirect-ELISA. Conversely, serum samples obtained from apparently healthy cattle were exclusively screened using indirect ELISA. None of the samples tested could detect C. burnetii in PCR assays, while 13.37% of serum samples were found to be seropositive in i-ELISA. Seropositivity noted among clinically healthy and those suffering from reproductive disorders were 12.76% and 16%, respectively, exhibiting a non-significant difference observed between these two categories. The obtained results suggested that the occurrence of coxiellosis did not differ significantly between clinically healthy animals and those with reproductive disorders; hence, in farms affected with C. burnetii infection, screening healthy and symptomatic animals is crucial to implement appropriate preventive measures.

Background & objectivesQ fever is an important zoonotic disease affecting humans as well as animals. The objective of this study was to assess the burden of Q fever in individuals with acute febrile illness, particularly those in close contact with animals. Various diagnostic methods were also evaluated in addition to clinical examination analysis and associated risk factors.MethodsIndividuals presenting with acute febrile illness who had animal exposure were enrolled (n=92) in this study. Serum samples were tested using IgG and IgM phase 2 enzyme linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA). The PCR targeting thecom1andIS1111genes was performed on blood samples. PCR amplicons were sequenced and phylogenetically analysed. Demographic data, symptoms, and risk factors were collected through a structured questionnaire.ResultsAmong individuals with acute febrile illness, 34.7 per cent (32 out of 92) were found to be infected withCoxiella burnetii.PCR exhibited the highest sensitivity among the diagnostic methods employed. The most common clinical manifestations included headache, chills, arthralgia, and fatigue. Individuals engaged in daily livestock-rearing activities were found to be at an increased risk of infection.Interpretation & conclusionsQ fever is underdiagnosed due to its varied clinical presentations, diagnostic complexities, and lack of awareness. This study underscores the importance of regular screening for Q fever in individuals with acute febrile illness, particularly those with animal exposure. Early diagnosis and increased awareness among healthcare professionals are essential for the timely management and prevention of chronic complications associated with Q fever.

The present study assessed the presence of ESBL-producing Escherichia coli in livestock farm wastewater (LFWW), hospital wastewater (HWW), and natural water sources (NWS) from five districts (Prayagraj, Mirzapur, Varanasi, Sonbhadra, and Jaunpur) of eastern parts of Uttar Pradesh, India (n = 134). Phenotypic ESBL production among cefotaxime-resistant E. coli isolates (91.29

Coxiella burnetii is a gram-negative, intracellular bacterium that shows extremely high infectivity (at an infection dose of down to a single organism) and causes diseases in a range of hosts including humans. Transmitted mainly through aerosols and ingestion of fomites from infected animals, human coxiellosis may manifest as acute Q fever (fever, headache, myalgia, arthralgia and diarrhea), chronic Q fever (endocarditis), Q fever during pregnancy (abortion) and Q fever fatigue syndrome (mental illness), depending on host immune status. As C. burnetii attacks monocytes and macrophages, which are important immune cells, and replicates within acidic phagolysosomes, it is not surprising that host innate and adaptive immune responses appear largely ineffective in the elimination of invading bacteria. The ability of C. burnetii to alter the structure of its lipopolysaccharide and to use other strategies further enhances its evasion of host defenses and its persistence within the host. Current approaches for diagnosing Q fever rely on the detection of C. burnetii DNA and/or specific antibodies in blood samples. The mainstay treatments for coxiellosis are doxycycline and hydroxychloroquine. Although one vaccine, Q-Vax, is commercially available for the prevention of human coxiellosis, additional research and development are warranted towards the design of innovative, tailor-made prophylaxes with increasing efficacy against coxiellosis in humans as well as animals.

This study attempted green synthesis of silver/zinc oxide nanocomposites (Ag/ZnO NCs) using methanolic extract of stem and leaves of Curcuma longa and its antibacterial efficacy against multi-drug-resistant (MDR) pathogens. UV-Vis spectroscopy suggested green synthesis of Ag/ZnO NCs which was verified by Fourier transform infrared spectroscopy, while thermogravimetric analysis with differential thermogravimetric analysis revealed its thermal stability between 400 degrees C and 700 degrees C. Powder X-ray diffraction results revealed irregular poly-crystalline morphology of Ag/ZnO NCs with certain extent of agglomeration, which was further confirmed by field emission-scanning electron microscopy and transmission electron microscopy. Further, minimum inhibitory concentration of 31.25 mu g/mL and minimum bactericidal concentration ranging from 62.50 to 125 mu g/ mL were observed against MDR strains of non-typhoidal Salmonella spp., enteroaggregative Escherichia coli and methicillin-resistant Staphylococcus aureus. This study demonstrated facile synthesis of Ag/ZnO NCs with promising antibacterial activity which could further be explored as potential antibiotic alternative.

Zeolite imidazole framework is one of the sub-classes of metal-organic frameworks which can enhance the antimicrobial activity of antibiotics against multi-drug resistant (MDR) bacteria. In this study, the authors report nanoscale dual metal cluster-based ZIF-8(Fe) to encapsulate citral. UV-DRS and FTIR spectra confirmed the encapsulation of citral, while PXRD illustrated an average particle size of 36 nm with uniform-sized nanoparticles. Drug encapsulation and loading efficiency of citral into ZIF-8(Fe) were found as 19.30 and 77.20