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1.
Food Chem ; 446: 138805, 2024 Jul 15.
Article in English | MEDLINE | ID: mdl-38422639

ABSTRACT

Non-specific binding in fluorescence resonance energy transfer (FRET) remains a challenge in foodborne pathogen detection, resulting in interference of high background signals. Herein, we innovatively reported a dual-mode FRET sensor based on a "noise purifier" for the ultrasensitive quantification of Escherichia coli O157:H7 in food. An efficient FRET system was constructed with polymyxin B-modified nitrogen-sulfur co-doped graphene quantum dots (N, S-GQDs@PMB) as donors and aptamer-modified yellow carbon dots (Y-CDs@Apt) as acceptors. Magnetic multi-walled carbon nanotubes (Fe@MWCNTs) were employed as a "noise purifier" to reduce the interference of the fluorescence background. Under the background purification mode, the sensitivity of the dual-mode signals of the FRET sensor has increased by an order of magnitude. Additionally, smartphone-assisted colorimetric analysis enabled point-of-care detection of E. coli O157:H7 in real samples. The developed sensing platform based on a "noise purifier" provides a promising method for ultrasensitive on-site testing of trace pathogenic bacteria in various foodstuffs.


Subject(s)
Nanotubes, Carbon , Quantum Dots , Fluorescence , Smartphone , Escherichia coli , Quantum Dots/chemistry , Point-of-Care Testing
2.
Int J Food Microbiol ; 414: 110615, 2024 Apr 02.
Article in English | MEDLINE | ID: mdl-38325260

ABSTRACT

Burkholderia gladiolus (B. gladiolus) is foodborne pathogenic bacteria producing bongkrekic acid (BA), which causes food poisoning and has a mortality rate of up to 40 % or more. However, no drugs have been reported in the literature for the prevention and treatment of this infection. In this study, a phage was identified to control B. gladiolus. The novel phage vB_BglM_WTB (WTB), which lyse B. gladiolus with high efficiency, was isolated from sewage of Huaihe Road Throttle Well Sewage Treatment Plant in Hefei. Transmission electron microscopy showed that WTB had an icosahedral head (69 ± 2 nm) and a long retractable tail (108 ± 2 nm). Its optimal temperature and pH ranges to control B. gladiolus were 25 °C -65 °C and 3-11 respectively. The phage WTB was identified as a linear double-stranded DNA phage of 68, 541 bp with 60.04 % G + C content, with a long latent period of 60 min. Phylogenetic analysis and comparative genetic analysis indicated that phage WTB has low identity (<50 %) with other phages, with the highest similarity to Burkholderia phage Maja (25.7 %), which showed that it does not belong to any previous genera recognized by the International Committee on Taxonomy of Viruses (ICTV) and was a candidate for a new genus within the Caudoviricetes. We have submitted a new proposal to ICTV to create a new genus, Bglawtbvirus. No transfer RNA (tRNA), virulence associated and antibiotic resistance genes were detected in phage WTB. Experimental results indicated that WTB at 4 °C and 25 °C had excellent inhibition activity against B. gladiolus in the black fungus, with an inhibition efficiency of over 99 %. The amount of B. gladiolus in the black fungus was reduced to a minimum of 89 CFU/mL when treated by WTB at 25 °C for 2 h. The inhibition rate remained at 99.97 % even after 12 h. The findings showed that the phage WTB could be applied as a food-cleaning agent for enhancing food safety and contributed to our understanding of phage biology and diversity.


Subject(s)
Bacteriophages , Burkholderia , Bacteriophages/genetics , Burkholderia/genetics , Sewage , Phylogeny , Genome, Viral , DNA, Viral/genetics , Fungi/genetics
3.
Food Microbiol ; 119: 104455, 2024 May.
Article in English | MEDLINE | ID: mdl-38225055

ABSTRACT

Cronobacter (seven species) can survive in dry powdered infant formula for a long time, but the thorough molecular mechanism of resistance to desiccation remains elusive. Here we examine the regulation mechanism of Cronobacter's tolerance to desiccation by the typical two-component system (TCS) EnvZ/OmpR. When exposed to desiccation conditions, Cronobacter showed higher survival than other pathogens, as well as significantly up-regulated expression of ompR and otsAB genes with markedly decreased survival of their mutants, suggesting their relationship with desiccation tolerance. OmpR directly binds to the promoter of trehalose biosynthesis operon otsBA, significantly enhancing their expression, and boosting the trehalose levels. The ompR-deletion in other six species further confirmed its positive regulation in desiccation tolerance. Our data present a hypothesis that EnvZ/OmpR increases intracellular trehalose levels against damage to the cells, which prompts Cronobacter to survive in desiccation conditions. This study reveals a universal molecular mechanism for desiccation resistance in Cronobacter species.


Subject(s)
Cronobacter , Humans , Infant , Cronobacter/genetics , Trehalose , Desiccation , Promoter Regions, Genetic , Bacterial Outer Membrane Proteins/genetics , Bacterial Outer Membrane Proteins/metabolism
4.
Biosens Bioelectron ; 248: 115991, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38169279

ABSTRACT

The effective combination of ultra-precise detection and on-demand sterilization stands out as one of the most valuable antifouling methods to combat pathogenic bacteria source and ensure the environment and food safety. Herein, an innovative "five birds one stone" aptasensor has been reported. It integrates magnetic separation, tri-modal precision detection, and efficient sterilization for monitoring of Staphylococcus aureus. Firstly, as a switch of the aptasensor, aptamer-modified potassium chloride-doped carbon dots (apt/KCl@CDs) could be adsorbed onto the surface of magnetic multi-walled carbon nanotube composites (M-MWCNTs) through π-π stacking, which could be replaced by the specific binding of the target bacteria to the aptamer. The mutual interference between the nanomaterials could be eliminated by this reverse magnetosorption strategy, enhancing the test sensitivity. In addition to the fluorescence properties, the peroxidase activity possessed by apt/KCl@CDs enables the conversion of the (3,3',5,5'-tetramethylbenzidine) TMB-H2O2 colorimetric system to a photothermal modal. Then, the ultra-precision detection in the assay was achieved by the fluorescence-colorimetric-photothermal tri-modal sensing from the formation of S. aureus-apt/KCl@CDs in the supernatant. Besides, the efficient sterilization could be ensured by adsorbing the apt/KCl@CDs on the surface of S. aureus, generating toxic •OH for direct attacking cells. This was the first report that was more beneficial for bacterial eradication. The detection limits of fluorescence, colorimetric and photothermal modals were 4.81 cfu/mL, 3.40 cfu/mL and 6.74 cfu/mL, respectively. The magnetic nanoplatform integrating tri-modal detection-sterilization meets the demand for highly sensitive and precise detection in different scenarios, providing immediate control for pathogens and broad application prospects.


Subject(s)
Anti-Infective Agents , Aptamers, Nucleotide , Biosensing Techniques , Staphylococcus aureus/chemistry , Hydrogen Peroxide , Biosensing Techniques/methods , Bacteria , Magnetic Phenomena , Limit of Detection , Aptamers, Nucleotide/chemistry
5.
Food Res Int ; 174(Pt 1): 113642, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37986541

ABSTRACT

Low-moisture foods (LMF) have arisen an increasing concern as vehicles of foodborne pathogens. Cronobacter genus, a class A pathogen in powdered infant formula (PIF), is crucial to the safety of LMF. Researchers have concentrated more on the bacterial survival caused by key hazardous factors, yet they often ignore the alteration of virulence properties in the surviving strains following rehydration of LMF mediated by the key factors. Our previous transcriptional profiling showed that luxS might participate in desiccation response. Herein, we further investigated the role of Cronobacter LuxS under desiccation stress by combining with the phenotypic and gene analysis between the Cronobacter parent and luxS mutant strains. Desiccation stress destructing assays confirmed that luxS can significantly enhance the resistance of Cronobacter towards desiccation. Our results also showed that cell hydrophobicity, aggregation, motility, the content of polysaccharide, and AI-2 synthesis pathway involved in luxS-mediated desiccation response. The luxS mutant strain exhibited higher swimming and swarming motility, more content of capsular polysaccharide, and more rapid of aggregation, but lower hydrophobicity than that of the wild-type strain, whereas desiccation stress would result in a opposite effect on these cell surface properties in ΔluxS during rehydration. Additionally, the comparation of gene expression profiles indicated that low moisture would trigger Cronobacter luxS to promote transport osmoprotectants by regulating the expression of proX, proW, and treC, and suppress the expression of cpsG associated with polysaccharide colanic acid. Notably, this study also discovered for the first time that the luxS-deficiency dramatically attenuated adhesion and invasion to intestinal and brain cells, but ΔluxS subjected to desiccation could aggravate the cell virulence instead. Therefore, thinking the alteration of toxicity caused by low-moisture, approach based on blocking the expression of the luxS gene to prevent Cronobacter in LMF needs to be adopted with caution.


Subject(s)
Cronobacter , Infant , Humans , Cronobacter/metabolism , Virulence/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Fluid Therapy , Polysaccharides
6.
Food Res Int ; 172: 113143, 2023 10.
Article in English | MEDLINE | ID: mdl-37689845

ABSTRACT

Cronobacter malonaticus (C. malonaticus) is a food-borne pathogen inducing severe infections both in infants and adults, and it could survive in dry powdered infant formula (PIF) for a long time, implying its strong tolerance to desiccation. However, the thorough molecular mechanism of resistance to desiccation remains elusive. When C. malonaticus was exposed to desiccation conditions (7, 15, and 30 d), transcriptomic analysis provided a universal adaptation strategy to withstand desiccation with the increased compatible solutes accumulation, activated stress resistance-related regulators, suppressed protein export and bacterial secretion system, and reduced other unessential survival functions including adhesion, invasion, virulence, and flagellar motility. Importantly, type VI secretion system (T6SS) genes exhibited significantly downregulated expressions, as well as markedly increased survival and viability of their mutants after desiccation treatment, revealing the negative regulation of T6SS in desiccation tolerance. Meanwhile, the decreased expressions of T6SS structure genes in other six species further confirmed the vital role of T6SS in desiccation tolerance of Cronobacter spp. Thus, our studies present a novel hypothesis of desiccation resistance in Cronobacter based on type VI secretion system inhibition, causing the reduction of macromolecule secretion such as effectors and hyperosmolality development within the cytomembrane, which allow Cronobacter to survive in desiccation.


Subject(s)
Cronobacter , Type VI Secretion Systems , Adult , Infant , Humans , Desiccation , Transcriptome , Cronobacter/genetics , Infant Formula
7.
Food Res Int ; 172: 113214, 2023 10.
Article in English | MEDLINE | ID: mdl-37689848

ABSTRACT

Contamination of infant formula with Cronobacter sakazakii (C. sakazakii) can cause fatal infections in neonates. Phages have emerged as promising antibacterial agents for food safety, but their effectiveness may be limited by thermal processing. In this study, we isolated 27 C. sakazakii phages from environmental water samples and selected LPCS28 due to its broad lysis spectrum. The titer of LPCS28 will not be significantly affected by heating at a temperature of 60 °C for one hour. In both reconstituted powdered infant formula (RPIF) and liquid milk, the pre-added LPCS28, after the thermal processing at 63 °C for 30 min, significantly inhibited the post-contaminated C. sakazakii (103 CFU/mL) and eventually reduced the number of C. sakazakii to below the limit of detection (<10 CFU/mL) within 9 h at 37 °C and significantly delayed the increase of bacterial concentration in the samples at 23 °C. The phylogenetic analysis revealed that LPCS28 belonged to a new genus, we proposed as Nanhuvirus, under the family Straboviridae. These findings suggest that phage LPCS28 is a promising biological control agent for pathogenic C. sakazakii in the dairy industry.


Subject(s)
Bacteriophages , Cronobacter sakazakii , Humans , Infant , Infant, Newborn , Animals , Milk , Infant Formula , Phylogeny , Powders
8.
Foods ; 12(13)2023 Jun 30.
Article in English | MEDLINE | ID: mdl-37444298

ABSTRACT

This research investigates the antibacterial potential of plant essential oil components including thymol, carvacrol, citral, cinnamaldehyde, limonene, and ß-pinene against Salmonella Enteritidis (S. Enteritidis). Through the determination of minimum inhibitory concentration, three kinds of natural antibacterial agents with the best inhibitory effect on S. Enteritidis were determined, namely thymol (128 µg/mL), carvacrol (256 µg/mL), and cinnamaldehyde (128 µg/mL). Physical, chemical, microbial, and sensory characteristics were regularly monitored on days 0, 2, 4, and 6. The findings of this study reveal that both thymol at MIC of 128 µg/mL and carvacrol at MIC of 256 µg/mL not only maintained the sensory quality of chicken, but also decreased the pH, moisture content, and TVB-N value. Additionally, thymol, carvacrol and cinnamaldehyde successfully inhibited the formation of S. Enteritidis biofilm, thereby minimizing the number of S. Enteritidis and the total aerobic plate count in chicken. Hence, thymol, carvacrol, and cinnamaldehyde have more effective inhibitory activities against S. Enteritidis, which can effectively prevent the spoilage of chicken and reduce the loss of its functional components.

9.
Article in English | MEDLINE | ID: mdl-37458925

ABSTRACT

Aspergillus fungi are widely used in the traditional fermentation of food products, so their safety risks and functions are worthy of investigation. In this study, one Aspergillus luchuensis YZ-1 isolated from Liubao tea was identified based on phylogenetic analyses of sequences of three genes coding for internal transcribed spacer 1 (ITS1), ß-tubulin (benA), and calmodulin (CaM). The results of hemolytic activity, DNase activity, cytotoxicity assay, and antibiotic resistance assay indicated that the strain is potentially safe. The excellent gastrointestinal fluid tolerance, acid tolerance, bile tolerance, auto-aggregation, co-aggregation, cell surface hydrophobicity, and adhesion to human colon adenocarcinoma (HT29) cell line were observed on analysis of the probiotic properties. Furthermore, the results of the antibacterial activity of A. luchuensis YZ-1 indicated that the strain had strong antagonistic effects against Gram-negative and Gram-positive bacteria as well as fungi. Simultaneously, the water extracts and 80% ethanolic extracts of A. luchuensis YZ-1 cells also showed strong ABTS, DPPH, and OH- scavenging ability. Taken together, our results suggest that A. luchuensis YZ-1 has desirable functional probiotic properties and can be proposed as a biocontrol agent in the food industry.

10.
Acta Biomater ; 168: 580-592, 2023 09 15.
Article in English | MEDLINE | ID: mdl-37451659

ABSTRACT

Healing bacterial chronic wounds caused by hyperglycemia is of great significance to protect the physical and mental health of diabetic patients. In this context, emerging chemodynamic therapy (CDT) and photothermal therapy (PTT) with broad antibacterial spectra and high spatiotemporal controllability have flourished. However, CDT was challenged by the near-neutral pH and inadequate H2O2 surrounding the chronic wound site, while PTT showed overheating-triggered side effects (e.g., damaging the normal tissue) and poor effects on thermotolerant bacterial biofilms. Therefore, we engineered an all-in-one glucose-responsive photothermal nanozyme, GOX/MPDA/Fe@CDs, consisting of glucose oxidase (GOX), Fe-doped carbon dots (Fe@CDs), and mesoporous polydopamine (MPDA), to efficiently treat chronic diabetic wound bacterial infections and eradicate biofilms without impacting the surrounding normal tissues. Specifically, GOX/MPDA/Fe@CDs produced a local temperature (∼ 45.0°C) to enhance the permeability of the pathogenic bacterium and its biofilm upon near-infrared (NIR) 808 nm laser irradiation, which was seized to initiate endogenous high blood glucose to activate the catalytic activity of GOX on the GOX/MPDA/Fe@CD surface to achieve the simultaneous self-supplying of H2O2 and H+, cascade catalyzing •OH production via a subsequent peroxidase-mimetic activity-induced Fenton/Fenton-like reaction. As such, the in vivo diabetic wound infected with methicillin-resistant Staphylococcus aureus was effectively healed after 12.0 days of treatment. This work was expected to provide an innovative approach to the clinical treatment of bacterially infected diabetic chronic wounds. STATEMENT OF SIGNIFICANCE: An all-in-one glucose-responsive photothermal nanozyme GOX/MPDA/Fe@CDs was constructed. Cascade nanozyme GOX/MPDA/Fe@CDs self-supply H2O2 and H+ to break H2O2 and pH limits to fight bacterial infections. Synergistic chemotherapy and photothermal therapy with nanozyme GOX/MPDA/Fe@CDs accelerates healing of biofilm-infected diabetic wounds.


Subject(s)
Diabetes Mellitus , Hyperglycemia , Methicillin-Resistant Staphylococcus aureus , Humans , Hydrogen Peroxide/pharmacology , Photothermal Therapy , Anti-Bacterial Agents/pharmacology , Carbon/pharmacology , Glucose , Glucose Oxidase/pharmacology , Nanotechnology
11.
Food Res Int ; 167: 112646, 2023 05.
Article in English | MEDLINE | ID: mdl-37087237

ABSTRACT

Aspergillus flavus contamination is common in various food and feed ingredients, and it poses to serious threats to human and animal health. Curcumin is a plant-derived polyphenol that exhibits antifungal activity. In this study, the antifungal effect of curcumin on A. flavus was evaluated, and the underlying mechanism was investigated. Curcumin effectively decreased aflatoxin B1 synthesis and suppressed A. flavus infection in peanut. Curcumin inhibited the mycelial growth and sporulation of A. flavus. Ergosterol biosynthesis in A. flavus was suppressed, and cell membrane permeability was enhanced. The pathogenicity of A. flavus was also reduced by curcumin treatment. Curcumin induced ROS burst in the hyphae of A. flavus, and those damages could be reversed by exogenous superoxide dismutase, suggesting that curcumin inhibited A. flavus possibly via inducing oxidative stress. These results indicate that curcumin has the potential to be used as a preservative to control A. flavus contamination in food and feedstuff.


Subject(s)
Aflatoxins , Curcumin , Humans , Aflatoxins/metabolism , Aspergillus flavus , Reactive Oxygen Species/metabolism , Curcumin/pharmacology , Curcumin/metabolism , Antifungal Agents/pharmacology , Antifungal Agents/metabolism
12.
Food Res Int ; 165: 112454, 2023 03.
Article in English | MEDLINE | ID: mdl-36869473

ABSTRACT

Thermal processing is the most frequently used method to destruct bacteria in food processing. However, insufficient thermal processing may lead to the outbreak of foodborne illness. This study combined thermal processing with thermostable phage to prevent food contamination. The thermostable phages were screened which can retain activity at 70 °C for 1 h. Among them, the polyvalent phage LPEK22 was obtained to lyse Escherichia coli and Salmonella enterica, especially several multi-drug resistant bacteria. In milk (liquid food matrix), LPEK22 significantly reduced the E. coli by 5.00 ± 0.18 log10 CFU/mL and S. enterica by 4.20 ± 0.23 log10 CFU/mL after thermal processing at 63 °C for 30 min. For beef sausage (solid food matrix), LPEK22 significantly reduced the E. coli by 2.34 ± 0.17 log10 CFU/cm2 and S. enterica by 1.54 ± 0.13 log10 CFU/cm2 after thermal processing at 66 °C for 90 s. Genome analysis revealed that LPEK22 was a novel phage with a unique tail spike protein belonging to the family of Ackermannviridae. LPEK22 did not contain lysogenic, drug-resistant, and virulent genes that may compromise the safety of food application. These results determined that LPEK22, a novel polyvalent Ackermannviridae phage, could combine with thermal processing to prevent drug-resistant E. coli and S. enterica both in vitro and in foods.


Subject(s)
Bacteriophages , Meat Products , Salmonella enterica , Cattle , Animals , Escherichia coli , Disease Outbreaks
13.
J Dairy Sci ; 106(3): 1576-1585, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36631321

ABSTRACT

Green hydrophobically modified butyrylated dextrin (BD) was used to modulate casein (CN). The CN/BD complex nanoparticles were formed at different CN-to-BD mass ratios based on a pH-driven technology. The interaction force, stability, and emulsifying properties of complex nanoparticles were investigated. The nanoparticles had a negative charge and a small particle size (160.03, 152.6, 155.9, 206.13, and 231.67 nm) as well as excellent thermal stability and environmental stability (pH 4.5, 5.5, 6.6, 7.5, 8.5, and 9.5; ionic strength, 50, 100, 200, and 500 mM). Transmission electron microscopy demonstrated the successful preparation of complex nanoparticles and their spherical shape. Fourier transform infrared spectroscopy, fluorescence spectroscopy, and dissociation analysis results showed that the main driving forces of formed CN/BD nanoparticles were hydrogen bonding and hydrophobic interaction. Furthermore, the CN/BD nanoparticles (CN/BD mass ratio, 1:1; weight/weight) exhibited the lowest creaming index, and optical microscopy showed that it has the most evenly dispersed droplets after 7 d of storage, which indicates that the CN/BD nanoparticles had excellent emulsifying properties. Butyrylated dextrin forms complex nanoparticles with CN through hydrogen bonding and hydrophobic interaction to endow CN with superior properties. The results showed that it is possible to use pH-driven technology to form protein-polysaccharide complex nanoparticles, which provides some information on the development of novel food emulsifiers based on protein-polysaccharide nanoparticles. The study provided significant information on the improvement of CN properties and the development of emulsions based on CN.


Subject(s)
Caseins , Nanoparticles , Animals , Caseins/chemistry , Dextrins , Emulsifying Agents , Emulsions/chemistry , Polysaccharides , Nanoparticles/chemistry , Particle Size
14.
Front Nutr ; 9: 1089890, 2022.
Article in English | MEDLINE | ID: mdl-36505266
15.
Front Immunol ; 13: 931871, 2022.
Article in English | MEDLINE | ID: mdl-36211338

ABSTRACT

Cronobacter has attracted considerable attention due to its association with meningitis and necrotizing enterocolitis (NEC) in newborns. Generally, lipopolysaccharide (LPS) facilitates bacterial translocation along with inflammatory responses as an endotoxin; however, the pathogenicity of Cronobacter LPS and the strategies to alleviate the toxicity were largely unknown. In this study, inflammatory responses were stimulated by intraperitoneal injection of Cronobacter malonaticus LPS into Sprague-Dawley young rats. Simultaneously, Bacteroides fragilis NCTC9343 were continuously fed through gavage for 5 days before or after injection of C. malonaticus LPS to evaluate the intervention effect of B. fragilis. We first checked the morphological changes of the ileum and colon and the intestinal microbiota and then detected the generation of inflammatory factors, including tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and interleukin-10 (IL-10) and the expression of Toll-like receptor 4 (TLR4), occludin, claudin-4, and iNOs. The results indicated that C. malonaticus LPS exacerbated intestinal infection by altering gut microbe profile, tight junction protein expression, and releasing inflammatory factors in a time- and dose-dependent manner. Intriguingly, treatment with B. fragilis obviously diminished the pathological injuries and expression of TLR4 caused by C. malonaticus LPS while increasing gut microbes like Prevotella-9. We note that Shigella, Peptoclostridium, and Sutterella might be positively related to C. malonaticus LPS infection, but Prevotella-9 was negatively correlated. The results suggested that the intestinal microbiota is an important target for the prevention and treatment of pathogenic injuries induced by C. malonaticus LPS.


Subject(s)
Cronobacter , Enterocolitis, Necrotizing , Gastrointestinal Microbiome , Animals , Bacteroides fragilis , Claudin-4/metabolism , Cronobacter/metabolism , Interleukin-10/metabolism , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Lipopolysaccharides/pharmacology , Occludin/metabolism , Rats , Rats, Sprague-Dawley , Toll-Like Receptor 4/metabolism , Tumor Necrosis Factor-alpha/metabolism
16.
J Dairy Sci ; 105(12): 9450-9462, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36207178

ABSTRACT

Foodborne pathogens detection is important to ensure food safety and human health. In this study, we designed a comet structure to rapidly and sensitively detect foodborne Listeria monocytogenes. This method combined isothermal sequence exchange amplification (SEA) and surface-enhanced Raman spectroscopy. Listeria monocytogenes DNA could be rapidly amplified at a constant temperature via SEA with a pair of modified primers, which rendered the precise thermal control instrumentation unnecessary. Efficient SEA amplification generated a large number of DNA duplexes that could be easily captured by streptavidin-modified magnetic bead and AuMB@Ag-isothiocyanate fluorescein antibody (anti-FITC). AuMB@Ag-anti-FITC was used as a signal probe, which generated a significant excitation signal at 1,616 cm-1 for quantitative detection and analysis. The results displayed sensitive detection of L. monocytogenes in cheese from 2.0 × 101 cfu/mL to 2.0 × 106 cfu/mL within 1.0 h with a detection limit of 7.8 cfu/mL. Furthermore, this comet structure displayed the desirable specificity as its specific primers and amplified DNA ends were attached to streptavidin-modified magnetic beads and AuMB@Ag-anti-FITC, respectively. We expected that the method devised would provide a promising new approach to screening for L. monocytogenes and guarantee the microbiological safety of dairy products.


Subject(s)
Cheese , Food Contamination , Listeria monocytogenes , Cheese/microbiology , DNA Primers/genetics , Food Microbiology , Listeria monocytogenes/isolation & purification , Spectrum Analysis, Raman , Streptavidin
17.
J Hazard Mater ; 439: 129582, 2022 10 05.
Article in English | MEDLINE | ID: mdl-35863223

ABSTRACT

Development of a simple and efficient dual-mode analytical technique with the built-in cross reference correction feature is benefit to achieve the highly accurate detection of the target pollutants and avoid the false-positive outputs in environmental media. Here, we synthesized a Fe-doped polydopamine (Fe@PDA)-based nanozyme with prominent peroxide-mimetic enzyme activity and high fluorescence emission ability. On this basis, we designed a dual-recognition strategy-driven fluorescence-colorimetric dual-mode detection platform, consisting of Listeria monocytogenes (L. monocytogenes) recognition aptamer-modified Fe@PDA (apt/Fe@PDA) and vancomycin-functionalized Fe3O4 (van/Fe3O4), for L. monocytogenes. Owing to van/Fe3O4-powered magnetic separation, there was a L. monocytogenes concentration-dependent fluorescence enhancement of apt/Fe@PDA for performing fluorescence assay in the precipitate. In this case, the prominent peroxide-mimetic enzyme activity of the residual apt/Fe@PDA in the precipitation could catalyze H2O2 to further oxidate colorless 3,3',5,5'-tetramethylbenzidine (TMB) into blue oxTMB, which displayed a L. monocytogenes concentration-dependent absorbance enhancement for carrying out colorimetric assay as well. As a result, a fluorescence-colorimetric dual-mode analytical platform was proposed to successfully detect the residual L. monocytogenes in real environmental media with acceptable results. This work showed the great prospects by integrating dual-recognition strategy into fluorescence nanozyme to develop efficient and reliable dual-mode analytical platforms for safeguarding environmental health.


Subject(s)
Colorimetry , Listeria monocytogenes , Colorimetry/methods , Hydrogen Peroxide , Indoles , Iron , Limit of Detection , Polymers
18.
Foods ; 11(15)2022 Jul 22.
Article in English | MEDLINE | ID: mdl-35892762

ABSTRACT

Lactiplantibacillus plantarum is a kind of extensively utilized probiotic species, which plays a critical role in the prevention of pathogenic bacteria and development of functional probiotics. Our group previously isolated one Lactiplantibacillus from Jiang Shui, a traditional Chinese fermented vegetable, which remarkably inhibited the growth of Aspergillus flavus. Herein, the safety of this isolate was assessed to ensure its application feasibility in food industry. Firstly, the phenotypic analyses including tolerance to low pH and bile salt, aggregation ability, and hemolytic activity detection, indicated the isolate could survive and colonize in the gastrointestinal tract, without hemolysin activity. The susceptibilities of the isolate to eight antibiotics and the absence of most resistance genes were demonstrated by agar disk diffusion and PCR, respectively. Furthermore, no mortality or toxicity was observed in mice by in vivo tests using gross autopsy, hematology, serum biochemistry, and HE-staining. Taken together, this study demonstrated the safety of Lactiplantibacillus plantarum WYH as a probiotic strain in terms of phenotypic analyses, absence of antimicrobial resistance and toxin-related genes, as well as mice toxicity test, while supported the prospect of applying isolate in suppression of fungal growth and mycotoxin biosynthesis.

19.
Mikrochim Acta ; 189(6): 233, 2022 05 27.
Article in English | MEDLINE | ID: mdl-35622176

ABSTRACT

Accurate and low-cost onsite assay of residual antibiotics in food and agriculture-related matrixes (e.g., milk) is of significant importance for evaluating and controlling food pollution risk. Herein, we employed hybrid Cu-doped-g-C3N4 nanozyme to engineer smartphone-assisted onsite visual sensor for reliable and precise reporting the levels of tetracycline (TC) residues in milk through π-π stacking-triggered blocking effect. Benefiting from the synergetic effects of Cu2+ and g-C3N4 nanosheet, Cu-doped-g-C3N4 nanocomposite exhibited an improved peroxidase-like activity, which could effectively catalyze H2O2 to oxidate colorless TMB into steel-blue product oxTMB. Interestingly, owing to the blocking effect caused by the π-π stacking interaction between TC tetraphenyl skeleton and Cu-doped-g-C3N4 nanozyme, the affinity of Cu-doped-g-C3N4 nanocomposite toward the catalytic substrates was remarkably blocked, resulting in a TC concentration-dependent fading of solution color. Using smartphone-assisted detection a simple, low-cost, reliable, and sensitive portable colorimetric sensor-based nanozyme for onsite visual monitoring the residual TC in milk was successfully developed with a detection limit of 86.27 nM. Of particular mention is that this detection limit is comparable to most other reported colorimetric methods and below most official allowable residue thresholds in milk matrixes. This work gave a novel insight to integrate two-dimensional (2D) artificial nanozymes-based π-π stacking-triggered blocking effect with smartphone-assisted detection for developing efficient and low-cost colorimetric point-of-care testing of the risk factors in food and agriculture-related matrixes.


Subject(s)
Colorimetry , Milk , Animals , Anti-Bacterial Agents/analysis , Colorimetry/methods , Hydrogen Peroxide/analysis , Milk/chemistry , Tetracycline/analysis
20.
J Hazard Mater ; 435: 128996, 2022 08 05.
Article in English | MEDLINE | ID: mdl-35487006

ABSTRACT

Bacterial biofilms have evoked worldwide attention owing to their serious threats to public health, but how to effectively eliminate bacterial biofilms still remains great challenges. Here, we rationally designed a novel and vigorous chitosan grafted Fe-doped-carbon dots (CS@Fe/CDs) as an efficient artificial nanozyme to combat rigid bacterial biofilms through the selective activation of Fenton-like reaction-triggered peroxidase-like catalytic activity and the synergistic antibacterial activity of CS. On the one hand, the peroxidase-like catalytic activity made CS@Fe/CDs catalyze H2O2 for producing hydroxyl radicals (•OH), resulting in efficient cleavage of extracellular DNA (eDNA). On the other hand, CS was capable of binding with the negatively charged cell membrane through electrostatic interaction, changing the cell membrane permeability and causing cell death within bacterial biofilms. Based on their synergistic effects, the fragments of bacterial biofilm and exposed bacteria were persistently eradicated. Remarkably, CS@Fe/CDs-based nanozyme not only enabled the effective destroying of gram-positive Staphylococcus aureus (S. aureus) biofilms, but also completely eliminated gram-negative Pseudomonas aeruginosa (P. aeruginosa) biofilms, showing great potential as a promising anti-biofilm agent against bacteria biofilms. This proposed synergistic strategy for bacterial biofilm eradication might offer a powerful modality to manage of bacterial biofilm fouling in food safety and environmental protection.


Subject(s)
Chitosan , Anti-Bacterial Agents/pharmacology , Bacteria , Biofilms , Carbon , Chitosan/pharmacology , Hydrogen Peroxide/pharmacology , Peroxidases , Pseudomonas aeruginosa , Staphylococcus aureus/physiology
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