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22 pages, 7118 KiB  
Article
A Novel Natural Chromogenic Visual and Luminescent Sensor Platform for Multi-Target Analysis in Strawberries and Shape Memory Applications
by Hebat-Allah S. Tohamy
Foods 2025, 14(16), 2791; https://doi.org/10.3390/foods14162791 - 11 Aug 2025
Viewed by 364
Abstract
Carboxymethyl cellulose (CMC) films, derived from sugarcane bagasse agricultural waste (SCB) incorporated with Betalains-nitrogen-doped carbon dots (Betalains-N–CQDs), derived from beet root waste (BR), offer a sustainable, smart and naked-eye sensor for strawberry packaging due to their excellent fluorescent and shape memory properties. These [...] Read more.
Carboxymethyl cellulose (CMC) films, derived from sugarcane bagasse agricultural waste (SCB) incorporated with Betalains-nitrogen-doped carbon dots (Betalains-N–CQDs), derived from beet root waste (BR), offer a sustainable, smart and naked-eye sensor for strawberry packaging due to their excellent fluorescent and shape memory properties. These CMC-Betalains-N–CQDs aim to enhance strawberry preservation and safety by enabling visual detection of common food contaminants such as bacteria, fungi and Pb(II). Crucially, the CMC-Betalains-N–CQD film also exhibits excellent shape memory properties, capable of fixing various shapes under alkaline conditions and recovering its original form in acidic environments, thereby offering enhanced physical protection for delicate produce like strawberries. Optical studies reveal the Betalains-N–CQDs’ pH-responsive fluorescence, with distinct emission patterns observed across various pH levels, highlighting their potential for sensing applications. Scanning Electron Microscopy (SEM) confirms the successful incorporation of Betalains-N–CQDs into the CMC matrix, revealing larger pores in the composite film that facilitate better interaction with analytes such as bacteria. Crucially, the CMC-Betalains-N–CQD film demonstrates significant antibacterial activity against common foodborne pathogens like Escherichia coli, Staphylococcus aureus, and Candida albicans, as evidenced by inhibition zones and supported by molecular docking simulations showing strong binding interactions with bacterial proteins. Furthermore, the film functions as a fluorescent sensor, exhibiting distinct color changes upon contact with different microorganisms and Pb(II) heavy metals, enabling rapid, naked-eye detection. The film also acts as a pH sensor, displaying color shifts (brown in alkaline, yellow in acidic) due to the betalains, useful for monitoring food spoilage. This research presents a promising, sustainable, and multifunctional intelligent packaging solution for enhanced food safety and extended shelf life. Full article
(This article belongs to the Section Food Packaging and Preservation)
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21 pages, 3124 KiB  
Article
Prevalence and Characterization of the Antimicrobial Resistance and Virulence Profiles of Staphylococcus aureus in Ready-to-Eat (Meat, Chicken, and Tuna) Pizzas in Mansoura City, Egypt
by Sara Amgad Elsalkh, Amira Ibrahim Zakaria, Samir Mohammed Abd-Elghany, Kálmán Imre, Adriana Morar and Khalid Ibrahim Sallam
Antibiotics 2025, 14(8), 817; https://doi.org/10.3390/antibiotics14080817 - 10 Aug 2025
Viewed by 474
Abstract
Introduction: Staphylococcus aureus is a high-priority foodborne pathogen contributing to several food poisoning outbreaks. Methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA), pose significant public health concerns due to their potential for serious illness, antibiotic resistance, and transmission within both healthcare and [...] Read more.
Introduction: Staphylococcus aureus is a high-priority foodborne pathogen contributing to several food poisoning outbreaks. Methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA), pose significant public health concerns due to their potential for serious illness, antibiotic resistance, and transmission within both healthcare and community settings. These bacteria can cause numerous infections, ranging from skin and soft tissue infections to life-threatening conditions like bloodstream infections, pneumonia, and endocarditis. Although several publications are concerned with Staphylococcus aureus contamination in ready-to-eat (RTE) food products, little published data is available about its prevalence in pizza, which is widely distributed and consumed worldwide. Methods: The current study is intended to determine the prevalence, virulence genes, and antimicrobial resistance profiles of S. aureus in three hundred ready-to-eat pizza samples (100 each of meat, chicken, and canned tuna pizzas) collected from different restaurants in Mansoura City, Egypt. The typical colonies on Baird–Parker selective agar supplemented with egg yolk tellurite emulsion were counted and further confirmed based on Gram staining, coagulase testing, catalase testing, carbohydrate fermentation, and thermostable nuclease production. The genomic DNA of the confirmed coagulase-positive isolates was prepared and subjected to PCR analyses for detecting the nuc gene, mecA (methicillin resistance gene), and vancomycin resistance gene (vanA), as well as six selected S. aureus virulence genes: sea, seb, sec, sed, hla, and tsst. The antimicrobial resistance profile of the S. aureus isolates was determined against 16 antimicrobial agents belonging to six classes using the agar disc diffusion method according to the Clinical and Laboratory Standards Institute guidelines (CLSI), except for oxacillin and vancomycin, which were assessed using the MIC test. Results: The results revealed that 56% (56/100), 56% (56/100), and 40% (40/100) of chicken, meat, and canned tuna pizzas were positive for S. aureus, with an overall prevalence of 50.7% (152/300). All 560 isolates (100%) were verified as S. aureus based on molecular confirmation of the nuc gene. Interestingly, 48.6% (272/560) and 8.6% (48/560) of the isolates tested were identified as methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA) through detection of mecA and vanA genes, respectively. Among the S. aureus isolates tested, the hla gene was detected in 87.1% (488/560), while the enterotoxin genes sea, seb, sec, and sed were identified in 50% (280/560), 78.6% (440/560), 9.8% (55/560), and 24.5% (137/560) of isolates, respectively. All recovered isolates (n = 560) were classified as multidrug-resistant and were resistant to penicillin, oxacillin, and ampicillin. Moreover, 77% (431/560), 24% (134/560), 8% (45/560), and 8.6% (48/560) of isolates were resistant to cefotaxime, ciprofloxacin, azithromycin, and vancomycin, respectively. Conclusions: The current study emphasizes that ready-to-eat pizza is highly contaminated with multidrug-resistant S. aureus, highlighting the urgent need for rationalizing antibiotic use in both veterinary and human medicine to prevent the transmission of resistant bacteria through the food chain. Additionally, strict adherence to good hygienic practices throughout all stages of the food chain is essential to minimize overall contamination and enhance food safety. Full article
(This article belongs to the Special Issue The Antimicrobial Resistance in the Food Chain)
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15 pages, 1339 KiB  
Article
Efficacy of Lactobacillus rhamnosus and Its Metabolites to Mitigate the Risk of Foodborne Pathogens in Hydroponic Nutrient Solution
by Esther Oginni, Robin Choudhury and Veerachandra Yemmireddy
Microorganisms 2025, 13(8), 1858; https://doi.org/10.3390/microorganisms13081858 - 8 Aug 2025
Viewed by 194
Abstract
Hydroponic nutrient solution (HNS) has been established as an ideal conduit for pathogen contamination and proliferation. This study evaluated the efficacy of lactic acid bacteria and their metabolites in mitigating the risk of foodborne pathogens in HNS when compared to conventional chemical treatments. [...] Read more.
Hydroponic nutrient solution (HNS) has been established as an ideal conduit for pathogen contamination and proliferation. This study evaluated the efficacy of lactic acid bacteria and their metabolites in mitigating the risk of foodborne pathogens in HNS when compared to conventional chemical treatments. Hoagland’s HNS were prepared according to the manufacturer’s instructions and inoculated with Salmonella Typhimurium, Escherichia coli 0157:H7, and Listeria innocua at 105 CFU/mL cell concentration. These nutrient solutions were subjected to treatment with various concentrations of Lactobacillus rhamnosus live cells, a cell-free extract (CFE) of L. rhamnosus metabolites, sodium hypochlorite and peroxyacetic acid at 22 ± 1 °C for up to 96 h using appropriate controls. The survived cells were enumerated on respective selective media at regular intervals. Additionally, the impact of these treatments on lettuce growth and the physico-chemical properties of HNS, such as pH, electrical conductivity, salinity, total dissolved solids, and % lactic acid content, were determined over 21 days using standard procedures. Both S. Typhimurium and E. coli O157: H7, when in combination with L. rhamnosus, remained stable in HNS over a 96 h period, while L. innocua showed a 3-log reduction. Whereas CFE treatment of HNS showed a significant reduction in Salmonella and E. coli O157: H7 (both undetectable after 96 h; LOD: <1 log CFU/mL). Interestingly, L. innocua levels remained stable after CFE treatment. PAA treatments at 12 mg/L notably reduced Salmonella and L. innocua growth, but not E. coli O157:H7. Lettuce plants in untreated control were significantly taller and heavier compared to those treated with CFE. These findings highlight the potential of biological interventions while emphasizing their limitations in hydroponic systems for pathogen control. Full article
(This article belongs to the Special Issue Feature Papers in Food Microbiology)
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20 pages, 1404 KiB  
Article
Bacteriophage PCSE1 as a Potential Strategy Against Salmonella Enteritidis in Liquid Egg Products
by Márcia Braz, Carla Pereira, Gabriela Matos, Jorge A. Saraiva, Carmen S. R. Freire and Adelaide Almeida
Antibiotics 2025, 14(8), 811; https://doi.org/10.3390/antibiotics14080811 - 8 Aug 2025
Viewed by 388
Abstract
Background/Objectives: The consumption of liquid egg products is rising. While thermal pasteurization improves safety and shelf life, it can affect product quality. Furthermore, egg products continue to cause many foodborne illnesses, especially those caused by Salmonella enterica subspecies enterica serovar Enteritidis (Salmonella [...] Read more.
Background/Objectives: The consumption of liquid egg products is rising. While thermal pasteurization improves safety and shelf life, it can affect product quality. Furthermore, egg products continue to cause many foodborne illnesses, especially those caused by Salmonella enterica subspecies enterica serovar Enteritidis (Salmonella Enteritidis). Bacteriophages (or phages) are an effective alternative to specifically fight foodborne bacteria. This study aimed to evaluate (i) the stability of phage vB_SeEM_UALMA_PCSE1 (PCSE1) under different conditions of temperature and pH; (ii) the effect of multiplicity of infection (MOI) and temperature on phage efficacy; (iii) the bactericidal effect of phage PCSE1 against S. Enteritidis in liquid whole eggs compared to thermal pasteurization; and (iv) the effect of both treatments on the physicochemical and functional properties of liquid whole eggs. Methods: For this, stability tests, bacterial growth inhibition assays in culture media and liquid eggs, and physicochemical and functional analyses were conducted. Results: Phage PCSE1 was (i) stable at pH 7 and 8, and at 4, 25, and 37 °C for 56 days; (ii) effectively prevented S. Enteritidis growth in TSB (reduction of 1.8, 4.5, and 4.5 log colony-forming units (CFU)/mL at 4, 10, and 25 °C, respectively, relative to the bacterial control); (iii) controlled S. Enteritidis in liquid whole eggs at 25 °C (reduction of 5.8 log CFU/mL relative to the bacterial control) comparable to pasteurization (reduction of 5.2 log CFU/mL); and (iv) preserved eggs’ properties, contrarily to pasteurization. Conclusions: These findings suggest PCSE1 is a promising strategy to fight S. Enteritidis in liquid egg products, though further studies on shelf-life are needed. Full article
(This article belongs to the Section Bacteriophages)
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17 pages, 701 KiB  
Article
Hydroethanolic Extracts of Raspberry (Rubus idaeus) Pomace as Ingredients of Functional Foods: Characterization and Effect of Gastrointestinal Digestion
by Ziva Vipotnik, Majda Golob and Alen Albreht
Plants 2025, 14(15), 2444; https://doi.org/10.3390/plants14152444 - 7 Aug 2025
Viewed by 374
Abstract
The extract of powdered raspberry pomace was characterized in terms of its phenolic profile and antioxidant and antimicrobial activity. Kuromanin, chlorogenic acid, protocatechuic acid, and pelargonidin-3-O-glucoside were found to be the major phenolic compounds, while the antioxidant activity of the extract [...] Read more.
The extract of powdered raspberry pomace was characterized in terms of its phenolic profile and antioxidant and antimicrobial activity. Kuromanin, chlorogenic acid, protocatechuic acid, and pelargonidin-3-O-glucoside were found to be the major phenolic compounds, while the antioxidant activity of the extract correlated positively with the total phenolic content (TPC), which was 472.9 ± 0.1 mg GAE/g dw. The extract also showed good antimicrobial activity against Gram-positive foodborne bacteria. More importantly, in vitro bioaccessibility of phenols from the raspberry pomace extract was 5-fold higher when the extract was incorporated into meringue cookies. Although the concentrations of anthocyanins, flavonoids, and tannins decreased after the oral, gastric, and intestinal phases of digestion, the TPC slightly increased as the compounds were released from the food matrix. The content of available phenolics was 4-fold lower in the case of a commercial raspberry colorant, demonstrating that the waste from raspberry pomace could serve as a valuable health-promoting ingredient for functional food formulations. Full article
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19 pages, 1579 KiB  
Article
Plasma-Treated Water Effect on Sporulating Bacillus cereus vs. Non-Sporulating Listeria monocytogenes Biofilm Cell Vitality
by Samantha Nestel, Robert Wagner, Mareike Meister, Thomas Weihe and Uta Schnabel
Appl. Microbiol. 2025, 5(3), 80; https://doi.org/10.3390/applmicrobiol5030080 - 5 Aug 2025
Viewed by 284
Abstract
Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have [...] Read more.
Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have compounding effects on the color, texture, flavor, and nutritional quality of the product or do not effectively reduce the pathogens that food can be exposed to. Some bacterial pathogens particularly possess traits and tactics that make them even more difficult to mitigate such as biofilm formation. Non-thermal plasma sanitation techniques, including plasma-treated water (PTW), have proven to be promising methods that significantly reduce pathogenic bacteria that food is exposed to. Published work reveals that PTW can effectively mitigate both gram-positive and gram-negative bacterial biofilms. This study presents a novel analysis of the differences in antimicrobial effects of PTW treatment between biofilm-forming gram-positive bacteria, commonly associated with foodborne illness, that are sporulating (Bacillus cereus) and non-sporulating (Listeria monocytogenes). After treatment with PTW, the results suggest the following hypotheses: (1) that the non-sporulating species experiences less membrane damage but a greater reduction in metabolic activity, leading to a possible viable but non-culturable (VBNC) state, and (2) that the sporulating species undergoes spore formation, which may subsequently convert into vegetative cells over time. PTW treatment on gram-positive bacterial biofilms that persist in food processing environments proves to be effective in reducing the proliferating abilities of the bacteria. However, the variance in PTW’s effects on metabolic activity and cell vitality between sporulating and non-sporulating species suggest that other survival tactics might be induced. This analysis further informs the application of PTW in food processing as an effective sanitation method. Full article
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13 pages, 1476 KiB  
Article
Molecular Detection and Antibiogram of Bacteria and Fungi in Table Eggs Under Different Storage Durations with Organoleptic Properties
by Md Shahab Uddin, Md Ahosanul Haque Shahid, Saiduzzaman, Marzia Rahman and K. H. M. Nazmul Hussain Nazir
Bacteria 2025, 4(3), 40; https://doi.org/10.3390/bacteria4030040 - 4 Aug 2025
Viewed by 303
Abstract
This study was undertaken to identify foodborne bacteria and fungi from different parts of eggs depending on their storage duration, organoleptic properties, total viable count, and antibiotic resistance profile. Thirty-two samples were randomly collected from commercial layer farms in Mymensingh. Following the protocol [...] Read more.
This study was undertaken to identify foodborne bacteria and fungi from different parts of eggs depending on their storage duration, organoleptic properties, total viable count, and antibiotic resistance profile. Thirty-two samples were randomly collected from commercial layer farms in Mymensingh. Following the protocol of sample preparation, outer-surface and inner-content samples were streaked onto various selective media. Isolation and identification were carried out by observing Gram staining and biochemical properties. Molecular detection was confirmed through a PCR assay using specific primers for Salmonella spp., E. coli, Staphylococcus spp., and fungus (Simplicillium spp. and Saccharomyces spp.). To determine the antibiotic resistance profile, the disk diffusion method was followed against nine antibiotic disks. The isolation rate of E. coli, Salmonella spp., and Staphylococcus spp. was 53.13%, 40.63%, and 40.63%, respectively, in the outer eggshell and 15.63%, 25%, and 15.63%, respectively, in the inner content of the eggs. Regarding the fungus content (yeast and mold), 100% was obtained in the outer eggshell, whereas there was an absence of fungus in the inner content. It was observed that all the isolates of E. coli, Salmonella spp., and Staphylococcus spp. were highly sensitive to either Ciprofloxacin or Levofloxacin and extremely resistant to Amoxicillin or Azithromycin drug disks or both. The data also shows that storage duration had a proportional relationship with TVC and an inversely proportional relationship with organoleptic properties. This study indicates that eggs harbor multidrug-resistant foodborne bacteria, which might constitute a public health hazard if these antibiotic-resistant bacteria are transferred to humans. Full article
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13 pages, 1944 KiB  
Article
A Sliding Microfluidic Chip-Integrated Colorimetric Biosensor Using MnO2 Nanoflowers for Rapid Salmonella Detection
by Yidan Niu, Juntao Jiang, Xin Zhi, Jiahui An and Yuhe Wang
Micromachines 2025, 16(8), 904; https://doi.org/10.3390/mi16080904 - 31 Jul 2025
Viewed by 318
Abstract
Rapid screening of foodborne pathogens is critical for food safety, yet current detection techniques often suffer from low efficiency and complexity. In this study, we developed a sliding microfluidic colorimetric biosensor for the fast, sensitive, and multiplex detection of Salmonella. First, the [...] Read more.
Rapid screening of foodborne pathogens is critical for food safety, yet current detection techniques often suffer from low efficiency and complexity. In this study, we developed a sliding microfluidic colorimetric biosensor for the fast, sensitive, and multiplex detection of Salmonella. First, the target bacteria were specifically captured by antibody-functionalized magnetic nanoparticles in the microfluidic chip, forming magnetic bead–bacteria complexes. Then, through motor-assisted sliding of the chip, manganese dioxide (MnO2) nanoflowers conjugated with secondary antibodies were introduced to bind the captured bacteria, generating a dual-antibody sandwich structure. Finally, a second sliding step brought the complexes into contact with a chromogenic substrate, where the MnO2 nanoflowers catalyzed a colorimetric reaction, and the resulting signal was used to quantify the Salmonella concentration. Under optimized conditions, the biosensor achieved a detection limit of 10 CFU/mL within 20 min. In spiked pork samples, the average recovery rate of Salmonella ranged from 94.9% to 125.4%, with a coefficient of variation between 4.0% and 6.8%. By integrating mixing, separation, washing, catalysis, and detection into a single chip, this microfluidic biosensor offers a user-friendly, time-efficient, and highly sensitive platform, showing great potential for the on-site detection of foodborne pathogens. Full article
(This article belongs to the Section B1: Biosensors)
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36 pages, 7948 KiB  
Review
Advancing Food Safety Surveillance: Rapid and Sensitive Biosensing Technologies for Foodborne Pathogenic Bacteria
by Yuerong Feng, Jiyong Shi, Jiaqian Liu, Zhecong Yuan and Shujie Gao
Foods 2025, 14(15), 2654; https://doi.org/10.3390/foods14152654 - 29 Jul 2025
Viewed by 751
Abstract
Foodborne pathogenic bacteria critically threaten public health and food industry sustainability, serving as a predominant trigger of food contamination incidents. To mitigate these risks, the development of rapid, sensitive, and highly specific detection technologies is essential for early warning and effective control of [...] Read more.
Foodborne pathogenic bacteria critically threaten public health and food industry sustainability, serving as a predominant trigger of food contamination incidents. To mitigate these risks, the development of rapid, sensitive, and highly specific detection technologies is essential for early warning and effective control of foodborne diseases. In recent years, biosensors have gained prominence as a cutting-edge tool for detecting foodborne pathogens, owing to their operational simplicity, rapid response, high sensitivity, and suitability for on-site applications. This review provides a comprehensive evaluation of critical biorecognition elements, such as antibodies, aptamers, nucleic acids, enzymes, cell receptors, molecularly imprinted polymers (MIPs), and bacteriophages. We highlight their design strategies, recent advancements, and pivotal contributions to improving detection specificity and sensitivity. Additionally, we systematically examine mainstream biosensor-based detection technologies, with a focus on three dominant types: electrochemical biosensors, optical biosensors, and piezoelectric biosensors. For each category, we analyze its fundamental principles, structural features, and practical applications in food safety monitoring. Finally, this review identifies future research priorities, including multiplex target detection, enhanced processing of complex samples, commercialization, and scalable deployment of biosensors. These advancements are expected to bridge the gap between laboratory research and real-world food safety surveillance, fostering more robust and practical solutions. Full article
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21 pages, 1797 KiB  
Article
Surface Functionalization of Cellulose-Based Packaging with a New Antimicrobial Decapeptide: A Sustainable Solution to Improve the Quality of Meat Products
by Bruna Agrillo, Rosa Luisa Ambrosio, Valeria Vuoso, Emanuela Galatola, Marta Gogliettino, Monica Ambrosio, Rosarita Tatè, Aniello Anastasio and Gianna Palmieri
Foods 2025, 14(15), 2607; https://doi.org/10.3390/foods14152607 - 24 Jul 2025
Viewed by 457
Abstract
The need for renewable and eco-friendly materials is driving the increasing demand for biobased polymers for food applications, with cellulose emerging as a promising option due to its degradability and environmental sustainability. Therefore, in the present study, a strategy to obtain cellulose-based materials [...] Read more.
The need for renewable and eco-friendly materials is driving the increasing demand for biobased polymers for food applications, with cellulose emerging as a promising option due to its degradability and environmental sustainability. Therefore, in the present study, a strategy to obtain cellulose-based materials with antimicrobial properties was explored by using a selected antimicrobial peptide named RKT1, which was stably and efficiently tethered to cellulose films via physical adsorption, harnessing the high number of functional groups on the polymeric surface. Firstly, the peptide, identified among the previous or new projected compounds, was structurally and functionally characterized, evidencing high conformational stability under a wide range of environmental conditions and efficient antibacterial activity against the foodborne pathogens Escherichia coli, Salmonella Typhimurium, and Listeria monocytogenes and the spoilage bacteria Enterococcus and Pseudomonas koreensis, all isolated from meat products. Moreover, in an extended application, the RKT1-activated cellulose films were tested in vivo on beef carpaccio. The results supported their effectiveness in increasing the shelf life of carpaccio by least two days without affecting its organoleptic properties. Therefore, RKT1, physically adsorbed on cellulose, still retains its activity, and the newly generated biopolymers show potential for use as a green food packaging material. Full article
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14 pages, 2465 KiB  
Article
Polymerase Chain Reaction-Lateral Flow Strip for Detecting Escherichia coli and Salmonella enterica Harboring blaCTX-M
by Rujirat Hatrongjit, Sumontha Chaisaeng, Kulsatree Sitthichotthumrong, Parichart Boueroy, Peechanika Chopjitt, Ratchadaporn Ungcharoen and Anusak Kerdsin
Antibiotics 2025, 14(8), 745; https://doi.org/10.3390/antibiotics14080745 - 24 Jul 2025
Viewed by 421
Abstract
Background: Salmonella enterica and Escherichia coli are common foodborne pathogens of global concern, particularly due to their antimicrobial resistance, notably to cephalosporins. Objective: This study aimed to evaluate a polymerase chain reaction-based lateral flow strip (PCR-LFS) assay for the detection of Salmonella [...] Read more.
Background: Salmonella enterica and Escherichia coli are common foodborne pathogens of global concern, particularly due to their antimicrobial resistance, notably to cephalosporins. Objective: This study aimed to evaluate a polymerase chain reaction-based lateral flow strip (PCR-LFS) assay for the detection of Salmonella spp. and E. coli harboring the blaCTX-M gene, which confers resistance to third-generation cephalosporins. Methods: Two duplex PCRs (dPCR) were established to detect E. coli-harboring blaCTX-M (set 1) and Salmonella-harboring blaCTX-M (set 2). 600 bacterial isolates and raw pork mince spiked with blaCTX-M-harboring E. coli and Salmonella were used to evaluated. Results: Both dPCR assays successfully detected blaCTX-M-positive E. coli or Salmonella strains, while strains lacking the gene showed no amplification. Non-E. coli and non-Salmonella strains were PCR-negative unless they carried blaCTX-M. The dPCR-LFS showed 100% validity including accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for both E. coli or Salmonella spp. harboring or lacking blaCTX-M. The assay accurately detected target strains without cross-reactivity with other bacteria or antimicrobial resistance genes. Cohen’s Kappa coefficient indicated perfect agreement (κ = 1), reflecting the high reliability of the dPCR-LFS. The assay could detect as low as 25 CFU/mL for blaCTX-M-positive E. coli and 40 CFU/mL for blaCTX-M-positive Salmonella in spiked raw pork mince. Conclusions: This assay is rapid, easy to interpret, and suitable for large-scale screening in surveillance programs. Full article
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12 pages, 2083 KiB  
Article
Modification of Silver-Loaded Biodegradable Polymer Nanoparticles with Bacterial Membrane Vesicles for Treating Intracellular Bacterial Infections
by Wei Xu, Sayo Maruyama and Takuro Niidome
Materials 2025, 18(15), 3470; https://doi.org/10.3390/ma18153470 - 24 Jul 2025
Viewed by 270
Abstract
Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen capable of surviving and replicating within macrophages, which causes foodborne diseases such as gastroenteritis. To develop a strategy against intracellular bacteria in macrophages, we designed silver-loaded biodegradable polymer nanoparticles functionalized [...] Read more.
Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen capable of surviving and replicating within macrophages, which causes foodborne diseases such as gastroenteritis. To develop a strategy against intracellular bacteria in macrophages, we designed silver-loaded biodegradable polymer nanoparticles functionalized with S. Typhimurium membrane vesicles (MVs). Silver nanoparticles (Ag NPs) were initially encapsulated within biodegradable poly(lactic-co-glycolic) nanoparticles (Ag-P NPs), which were then surface-modified with polyethyleneimine to form Ag-PP NPs. These were subsequently fused with S. Typhimurium MVs via a sonication method to generate Ag-PP@MV NPs. The resulting MV-coated nanoparticles displayed a similar protein profile to that of native MVs and exhibited antibacterial activity against intracellular S. Typhimurium. Notably, the enhanced cellular uptake of the MV-modified NPs contributed to their intracellular bactericidal efficacy. This study highlights MV modification as a promising strategy to improve NP delivery to macrophages for treating persistent intracellular infections. Full article
(This article belongs to the Section Biomaterials)
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19 pages, 7706 KiB  
Article
Genomic Insights into Vaccinium spp. Endophytes B. halotolerans and B. velezensis and Their Antimicrobial Potential
by Ingrida Mažeikienė, Birutė Frercks, Monika Kurgonaitė, Neringa Rasiukevičiūtė and Irena Mačionienė
Int. J. Mol. Sci. 2025, 26(14), 6677; https://doi.org/10.3390/ijms26146677 - 11 Jul 2025
Cited by 1 | Viewed by 314
Abstract
Plant microbiota contributes to nutrient absorption, and the production of hormones and vitamins, and plays a crucial role in responding to environmental stress. We hypothesized that Vaccinium spp. harbour a unique microbiota that enables them to coexist in extreme environments such as saline, [...] Read more.
Plant microbiota contributes to nutrient absorption, and the production of hormones and vitamins, and plays a crucial role in responding to environmental stress. We hypothesized that Vaccinium spp. harbour a unique microbiota that enables them to coexist in extreme environments such as saline, nutrient-poor, and waterlogged conditions. Upon examining Bacillus spp. endophytes isolated from blueberries, cranberries and lingonberries in vitro, we identified B. halotolerans (Bil-LT1_1, Bil-LT1_2) and B. velezensis (Cran-LT1_8, Ling-NOR4_15) strains that inhibit the growth of five pathogenic fungi and five foodborne bacteria. Whole-genome sequencing provided insights into genome organization and plasticity, helping identify mobile elements and genes potentially acquired through horizontal gene transfer. Functional annotation identified genes associated with plant colonization, stress tolerance, biocontrol activity, and plant growth promotion. Comparative genomic analyses revealed key biosynthetic gene clusters (BGCs) responsible for producing antifungal metabolites, including lipopeptides and polyketides. Genes supporting plant nutrition, growth, and environmental adaptation were present also in these strains. Notably, isolated endophytes exhibited particularly high levels of genomic plasticity, likely due to horizontal gene transfer involving gene ontology (GO) pathways related to survival in polymicrobial and foreign environments. Full article
(This article belongs to the Special Issue Microbial Omics: Decoding Microbial Life)
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23 pages, 3213 KiB  
Article
Multidrug Resistance and Virulence Traits of Salmonella enterica Isolated from Cattle: Genotypic and Phenotypic Insights
by Nada A. Fahmy, Sumin Karna, Angel Bhusal, Ajran Kabir, Erdal Erol and Yosra A. Helmy
Antibiotics 2025, 14(7), 689; https://doi.org/10.3390/antibiotics14070689 - 8 Jul 2025
Viewed by 874
Abstract
Background/Objective: Non-typhoidal Salmonella is a leading cause of foodborne illness worldwide and presents a significant One Health concern due to zoonotic transmission. Although antibiotic therapy remains a standard approach for treating salmonellosis in severe cases in animals, the widespread misuse of antibiotics has [...] Read more.
Background/Objective: Non-typhoidal Salmonella is a leading cause of foodborne illness worldwide and presents a significant One Health concern due to zoonotic transmission. Although antibiotic therapy remains a standard approach for treating salmonellosis in severe cases in animals, the widespread misuse of antibiotics has contributed to the emergence of multidrug-resistant (MDR) Salmonella strains. This study provides insights into the genotypic and phenotypic characteristics among Salmonella isolates from necropsied cattle. Methods: A total of 1008 samples were collected from necropsied cattle. Salmonella enterica subspecies were identified by MALDI-TOF MS and subsequently confirmed by serotyping. The biofilm-forming ability of the isolated bacteria was assessed using a crystal violet assay. The motility of the isolates was assessed on soft agar plates. Additionally, the antimicrobial resistance genes (ARGs) and virulence genes were investigated. Antimicrobial resistance patterns were investigated against 19 antibiotics representing 9 different classes. Results:Salmonella species were isolated and identified in 27 necropsied cattle. Salmonella Dublin was the most prevalent serotype (29.6%). Additionally, all the isolates were biofilm producers at different levels of intensity, and 96.3% of the isolates exhibited both swarming and swimming motility. Furthermore, virulence genes, including invA, hilA, fimA, and csgA, were detected in all the isolates. The highest resistance was observed to macrolides (azithromycin and clindamycin) (100%), followed by imipenem (92.6%), and chloramphenicol (85.2%). All isolates were multidrug-resistant, with a multiple antibiotic resistance (MAR) index ranging between 0.32 and 0.74. The aminoglycoside resistance gene aac(6′)-Ib was detected in all the isolates (100%), whereas the distribution of other antimicrobial resistance genes (ARGs) varied among the isolates. Conclusions: The increasing prevalence of MDR Salmonella poses a significant public health risk. These resistant strains can reduce the effectiveness of standard treatments and elevate outbreak risks. Strengthening surveillance and regulating antibiotic use in livestock are essential to mitigating these threats. Full article
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14 pages, 428 KiB  
Review
Potential Future Applications of Postbiotics in the Context of Ensuring Food Safety and Human Health Improvement
by Zorica Tomičić, Ljubiša Šarić and Ružica Tomičić
Antibiotics 2025, 14(7), 674; https://doi.org/10.3390/antibiotics14070674 - 3 Jul 2025
Cited by 2 | Viewed by 840
Abstract
Postbiotics are defined as non-viable metabolites or compounds produced by probiotic microorganisms with significant impact on human health. The growing interest in postbiotics is supported by numerous studies due to their additional benefits over probiotics that show positive outcomes for specific conditions, as [...] Read more.
Postbiotics are defined as non-viable metabolites or compounds produced by probiotic microorganisms with significant impact on human health. The growing interest in postbiotics is supported by numerous studies due to their additional benefits over probiotics that show positive outcomes for specific conditions, as well as their application as biopreservatives in the food industry. Their potential in functional foods and therapeutic applications is increasingly recognized as they exhibit stability, safety, and diverse biological activities. As for their most important biological roles, postbiotics have been shown to have effective anti-inflammatory, antimicrobial, antioxidant, and anticancer properties, in addition to reducing food allergies. The application of postbiotics in functional foods contributes to improving intestinal health and reducing the risk of foodborne diseases. The concept of postbiotics is relatively new in the food industry. They offer a promising alternative to conventional food preservatives due to their ability to inhibit pathogenic bacteria and extend shelf life. Considering the diversity of postbiotic compounds and their significant biological activities, this review presents and discusses the mechanisms of action and future trends in their application in the food industry and their impact on human health. Increasing research and development in the production and formulation of postbiotics will play a key role in the upward trajectory of the market. Full article
(This article belongs to the Special Issue Bioactive Natural Products in Antimicrobial Resistance Management)
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