Antibacterial Activity and Mechanisms of Action of Inorganic Nanoparticles against Foodborne Bacterial Pathogens: A Systematic Review.

Foodborne disease outbreaks due to bacterial pathogens and their toxins have become a serious concern for global public health and security. Finding novel antibacterial agents with unique mechanisms of action against the current spoilage and foodborne bacterial pathogens is a central strategy to overcome antibiotic resistance. This study examined the antibacterial activities and mechanisms of action of inorganic nanoparticles (NPs) against foodborne bacterial pathogens. The articles written in English were recovered from registers and databases (PubMed, ScienceDirect, Web of Science, Google Scholar, and Directory of Open Access Journals) and other sources (websites, organizations, and citation searching). "Nanoparticles," "Inorganic Nanoparticles," "Metal Nanoparticles," "Metal-Oxide Nanoparticles," "Antimicrobial Activity," "Antibacterial Activity," "Foodborne Bacterial Pathogens," "Mechanisms of Action," and "Foodborne Diseases" were the search terms used to retrieve the articles. The PRISMA-2020 checklist was applied for the article search strategy, article selection, data extraction, and result reporting for the review process. A total of 27 original research articles were included from a total of 3,575 articles obtained from the different search strategies. All studies demonstrated the antibacterial effectiveness of inorganic NPs and highlighted their different mechanisms of action against foodborne bacterial pathogens. In the present study, small-sized, spherical-shaped, engineered, capped, low-dissolution with water, high-concentration NPs, and in Gram-negative bacterial types had high antibacterial activity as compared to their counterparts. Cell wall interaction and membrane penetration, reactive oxygen species production, DNA damage, and protein synthesis inhibition were some of the generalized mechanisms recognized in the current study. Therefore, this study recommends the proper use of nontoxic inorganic nanoparticle products for food processing industries to ensure the quality and safety of food while minimizing antibiotic resistance among foodborne bacterial pathogens.

Multimodal Biosensing of Foodborne Pathogens.

Microbial foodborne pathogens present significant challenges to public health and the food industry, requiring rapid and accurate detection methods to prevent infections and ensure food safety. Conventional single biosensing techniques often exhibit limitations in terms of sensitivity, specificity, and rapidity. In response, there has been a growing interest in multimodal biosensing approaches that combine multiple sensing techniques to enhance the efficacy, accuracy, and precision in detecting these pathogens. This review investigates the current state of multimodal biosensing technologies and their potential applications within the food industry. Various multimodal biosensing platforms, such as opto-electrochemical, optical nanomaterial, multiple nanomaterial-based systems, hybrid biosensing microfluidics, and microfabrication techniques are discussed. The review provides an in-depth analysis of the advantages, challenges, and future prospects of multimodal biosensing for foodborne pathogens, emphasizing its transformative potential for food safety and public health. This comprehensive analysis aims to contribute to the development of innovative strategies for combating foodborne infections and ensuring the reliability of the global food supply chain.

Machine learning approach as an early warning system to prevent foodborne Salmonella outbreaks in northwestern Italy.

Salmonellosis, one of the most common foodborne infections in Europe, is monitored by food safety surveillance programmes, resulting in the generation of extensive databases. By leveraging tree-based machine learning (ML) algorithms, we exploited data from food safety audits to predict spatiotemporal patterns of salmonellosis in northwestern Italy. Data on human cases confirmed in 2015-2018 (n = 1969) and food surveillance data collected in 2014-2018 were used to develop ML algorithms. We integrated the monthly municipal human incidence with 27 potential predictors, including the observed prevalence of Salmonella in food. We applied the tree regression, random forest and gradient boosting algorithms considering different scenarios and evaluated their predictivity in terms of the mean absolute percentage error (MAPE) and R2. Using a similar dataset from the year 2019, spatiotemporal predictions and their relative sensitivities and specificities were obtained. Random forest and gradient boosting (R2 = 0.55, MAPE = 7.5%) outperformed the tree regression algorithm (R2 = 0.42, MAPE = 8.8%). Salmonella prevalence in food; spatial features; and monitoring efforts in ready-to-eat milk, fruits and vegetables, and pig meat products contributed the most to the models' predictivity, reducing the variance by 90.5%. Conversely, the number of positive samples obtained for specific food matrices minimally influenced the predictions (2.9%). Spatiotemporal predictions for 2019 showed sensitivity and specificity levels of 46.5% (due to the lack of some infection hotspots) and 78.5%, respectively. This study demonstrates the added value of integrating data from human and veterinary health services to develop predictive models of human salmonellosis occurrence, providing early warnings useful for mitigating foodborne disease impacts on public health.

Degradation of Toxins Derived from Foodborne Pathogens by Atmospheric-Pressure Dielectric-Barrier Discharge.

Foodborne diseases can be attributed not only to contamination with bacterial or fungal pathogens but also their associated toxins. Thus, to maintain food safety, innovative decontamination techniques for toxins are required. We previously demonstrated that an atmospheric-pressure dielectric-barrier discharge (APDBD) plasma generated by a roller conveyer plasma device is effective at inactivating bacteria and fungi in foods. Here, we have further examined whether the roller conveyer plasma device can be used to degrade toxins produced by foodborne bacterial pathogens, including aflatoxin, Shiga toxins (Stx1 and Stx2), enterotoxin B and cereulide. Each toxin was spotted onto an aluminum plate, allowed to dry, and then treated with APDBD plasma applied by the roller conveyer plasma device for different time periods. Assessments were conducted using a competitive enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results demonstrate a significant time-dependent decrease in the levels of these toxins. ELISA showed that aflatoxin B1 concentrations were reduced from 308.6 µg/mL to 74.4 µg/mL within 1 min. For Shiga toxins, Stx1 decreased from 913.8 µg/mL to 65.1 µg/mL, and Stx2 from 2309.0 µg/mL to 187.6 µg/mL within the same time frame (1 min). Enterotoxin B levels dropped from 62.67 µg/mL to 1.74 µg/mL at 15 min, and 1.43 µg/mL at 30 min, but did not display a significant decrease within 5 min. LC-MS/MS analysis verified that cereulide was reduced to below the detection limit following 30 min of APDBD plasma treatment. Taken together, these findings highlight that a range of foodborne toxins can be degraded by a relatively short exposure to plasma generated by an APDBD using a roller conveyer device. This technology offers promising advancements in food safety, providing a novel method to alleviate toxin contamination in the food processing industry.

Interference of gastrointestinal barriers with antibiotic susceptibility of foodborne pathogens: an in vitro case study of ciprofloxacin and tetracycline against Salmonella enterica and Listeria monocytogenes.

Minimum inhibitory concentrations (MIC) assays are often questioned for their representativeness. Especially when foodborne pathogens are tested, it is of crucial importance to also consider parameters of the human digestive system. Hence, the current study aimed to assess the inhibitory capacity of two antibiotics, ciprofloxacin and tetracycline, against Salmonella enterica and Listeria monocytogenes, under representative environmental conditions. More specifically, aspects of the harsh environment of the human gastrointestinal tract (GIT) were gradually added to the experimental conditions starting from simple aerobic lab conditions into an in vitro simulation of the GIT. In this way, the effects of parameters including the anoxic environment, physicochemical conditions of the GIT (low gastric pH, digestive enzymes, bile acids) and the gut microbiota were evaluated. The latter was simulated by including a representative consortium of selected gut bacteria species. In this study, the MIC of the two antibiotics against the relevant foodborne pathogens were established, under the previously mentioned environmental conditions. The results of S. enterica highlighted the importance of the anaerobic environment when conducting such studies, since the pathogen thrived under such conditions. Inclusion of physicochemical barriers led to exactly opposite results for S. enterica and L. monocytogenes since the former became more susceptible to ciprofloxacin while the latter showed lower susceptibility towards tetracycline. Finally, the inclusion of gut bacteria had a bactericidal effect against L. monocytogenes even in the absence of antibiotics, while gut bacteria protected S. enterica from the effect of ciprofloxacin.

Contaminação ambiental de uma agroindústria familiar de queijo colonial no sudoeste paranaense / Environmental contamination of a family colonial cheese agroindustry in the southwest of Parana

As falhas na higienização em um estabelecimento de alimentos podem refletir em problemas causando a contaminação ou deterioração do produto produzido. Esta pesquisa foi motivada por reclamações de consumidores informando que os queijos apresentaram fungos, mesmo estando dentro do prazo de validade e por solicitação do Serviço de Inspeção Municipal. O objetivo desta pesquisa foi avaliar a contaminação ambiental em uma agroindústria da agricultura familiar produtora de queijo colonial no Sudoeste Paranaense. Foram realizadas a contagem para aeróbios mesófilos em equipamentos e superfícies que entram em contato com o alimento e análise microbiológica ambiental de bolores e leveduras na sala de secagem dos queijos. A coleta foi realizada com método de esfregaço de suabe estéril para aeróbios mesófilos e semeadas em placas de Petri com Ágar Padrão de Contagem. Para a coleta ambiental foram expostas placas de Petri com ágar Saboraund durante 15 minutos. Os resultados demonstraram ausência de contaminação nas superfícies, mas foram encontrados bolores e leveduras de forma acentuada na sala de secagem dos queijos, o que pode contribuir para a deterioração do produto, diminuindo sua validade. Para minimizar as perdas por contaminação é necessário que o processo de higienização dos ambientes seja realizado de forma eficiente.

Failures in hygiene in a food establishment can result in problems causing contamination or deterioration of the product produced. This research was motivated by complaints from consumers reporting that the cheeses had mold, even though they were within their expiration date and at the request of the Municipal Inspection Service. This research was to evaluate environmental contamination in an agroindustry in the family farm producing colonial cheese in Southwest Paraná. For the microbiological assessment of environmental contamination, counting for mesophilic aerobes was carried out on equipment and surfaces that come into contact with food and, environmental microbiological analysis of molds and yeast in the cheese drying room. The collection was carried out using the sterile swab smear for mesophilic aerobes and seeded in Petri dishes with Counting Standard Agar. For environmental collection, sheets of Petri with Saboraund agar for 15 minutes. The results demonstrated absence of contamination on surfaces. But the presence of molds and yeasts in the drying room cheeses, which can contribute to the deterioration of the product and thus reduce the validity. To minimize losses due to contamination, it is It is necessary that the process of cleaning and disinfecting environments is carried out efficiently.

Higienização de ambiente em agroindústria familiar de embutidos cárneos / Environmental hygienization in family meat product

A higienização é um procedimento importante na indústria de alimentos e sua realização deve ocorrer rotineiramente para evitar que os alimentos sejam contaminados. Além disso, todos os manipuladores de alimentos devem receber treinamentos de modo a entender como ocorrem as contaminações e como evitá-las, para que não ocorra deterioração antecipada dos alimentos e para que não exponham os consumidores ao risco de doenças transmitidas por alimentos em caso de contaminação. Esta pesquisa avaliou o processo de higienização e sua eficiência em superfícies presentes em uma agroindústria da agricultura familiar produtora de embutidos cárneos. Apesar de ter instalações adequadas a agroindústria apresentava inadequações quanto aos produtos utilizados e a frequência inadequada para uma higienização eficiente. Foi realizada análise microbiológica das superfícies dos equipamentos para contagem de aeróbios mesófilos e notou-se uma elevada carga microbiana que indicou uma baixa eficiência no processo de higienização. Sugeriu-se melhorias na higiene ambiental associado à instrução dos colaboradores, para contribuir na promoção da qualidade dos produtos, aumento dos lucros e salvaguardando a saúde do consumidor.

Hygiene is an important procedure in the food industry, and its performance must occur routinely to prevent food from being contaminated. In addition, all food handlers must receive training in order to understand how contamination occurs and how to avoid it, so that there is no anticipated deterioration of food and that consumers are not exposed to the risk of foodborne diseases. in case of contamination by pathogenic microorganisms. Thus, this research evaluated the cleaning process and its efficiency on surfaces present in a family farming agroindustry that produces meat products, which despite having adequate facilities, had some difficulties such as product use and inadequate frequency for eficiente cleaning. After performing a microbiological analysis to count surface mesophilic aerobes, a high level of contamination was noted, relating to low efficiency in the cleaning process. Improvements in environmental hygiene are suggested, associated with the instruction of employees for the implementation of the Standard Operating Hygiene Procedure, promoting improvements in product quality, increasing profits and safeguarding consumer health.

CRISPR/Cas-based colorimetric biosensors: a promising tool for the diagnosis of bacterial foodborne pathogens in food products.

Some physical phenomena and various chemical substances newly introduced in nanotechnology have allowed scientists to develop valuable devices in the field of food sciences. Regarding such progress, the identification of foodborne pathogenic microorganisms is an imperative subject nowadays. These bacterial species have been found to cause severe health impacts after food ingestion and can result in high mortality in acute cases. The rapid detection of foodborne bacterial species at low concentrations is in high demand in recent diagnostics. CRISPR/Cas-mediated biosensors possess the potential to overcome several challenges in classical assays such as complex pretreatments, long turnaround time, and insensitivity. Among them, colorimetric nanoprobes based on the CRISPR strategy afford promising devices for POCT (point-of-care testing) since they can be visualized with the naked eye and do not require diagnostic apparatus. In this study, we briefly classify and discuss the working principles of the different CRISPR/Cas protein agents that have been employed in biosensors so far. We assess the current status of the CRISPR system, specifically focusing on colorimetric biosensing platforms. We discuss the utilization of each Cas effector in the detection of foodborne pathogens and examine the restrictions of the existing technology. The challenges and future opportunities are also indicated and addressed.

Food-Borne Biotoxin Neutralization in Vivo by Nanobodies: Current Status and Prospects.

Food-borne biotoxins from microbes, plants, or animals contaminate unclean, spoiled, and rotten foods, posing significant health risks. Neutralizing such toxins is vital for human health, especially after food poisoning. Nanobodies (Nbs), a type of single-domain antibodies derived from the genetic cloning of a variable domain of heavy chain antibodies (VHHs) in camels, offer unique advantages in toxin neutralization. Their small size, high stability, and precise binding enable effective neutralization. The use of Nbs in neutralizing food-borne biotoxins offers numerous benefits, and their genetic malleability allows tailored optimization for diverse toxins. As nanotechnology continues to evolve and improve, Nbs are poised to become increasingly efficient and safer tools for toxin neutralization, playing a pivotal role in safeguarding human health and environmental safety. This review not only highlights the efficacy of these agents in neutralizing toxins but also proposes innovative solutions to address their current challenges. It lays a solid foundation for their further development in this crucial field and propels their commercial application, thereby contributing significantly to advancements in this domain.

Optical sensing for real-time detection of food-borne pathogens in fresh produce using machine learning.

Contaminated fresh produce remains a prominent catalyst for food-borne illnesses, prompting the need for swift and precise pathogen detection to mitigate health risks. This paper introduces an innovative strategy for identifying food-borne pathogens in fresh produce samples from local markets and grocery stores, utilizing optical sensing and machine learning. The core of our approach is a photonics-based sensor system, which instantaneously generates optical signals to detect pathogen presence. Machine learning algorithms process the copious sensor data to predict contamination probabilities in real time. Our study reveals compelling results, affirming the efficacy of our method in identifying prevalent food-borne pathogens, including Escherichia coli (E. coli) and Salmonella enteric, across diverse fresh produce samples. The outcomes underline our approach's precision, achieving detection accuracies of up to 95%, surpassing traditional, time-consuming, and less accurate methods. Our method's key advantages encompass real-time capabilities, heightened accuracy, and cost-effectiveness, facilitating its adoption by both food industry stakeholders and regulatory bodies for quality assurance and safety oversight. Implementation holds the potential to elevate food safety and reduce wastage. Our research signifies a substantial stride toward the development of a dependable, real-time food safety monitoring system for fresh produce. Future research endeavors will be dedicated to optimizing system performance, crafting portable field sensors, and broadening pathogen detection capabilities. This novel approach promises substantial enhancements in food safety and public health.

Designing a Multiepitope Vaccine against the Foodborne Pathogenic Bacteria Listeria monocytogenes Using Subtractive Immunoinformatics Approaches.

BACKGROUND: Listeria monocytogenes, a Gram-positive bacterium, is a prominent foodborne pathogen that causes listeriosis and poses substantial health hazards worldwide. The continuing risk of listeriosis outbreaks underlies the importance of designing an effective prevention strategy and developing a robust immune response by reverse vaccinology approaches. This study aimed to provide a critical approach for developing a potent multiepitope vaccine against this foodborne disease. METHODS: A chimeric peptide construct containing 5 B-cell epitopes, 16 major histocompatibility complex I (MHC-I) epitopes, and 18 MHC-II epitopes were used to create a subunit vaccination against L. monocytogenes. The vaccine safety was evaluated by several online methods, and molecular docking was performed using ClusPro to determine the binding affinity. Immune simulation was performed using the C-ImmSimm server to demonstrate the immune response. RESULTS: The results validated the antigenicity, non-allergenicity, and nontoxicity of the chimeric peptide construct, confirming its suitability as a subunit vaccine. Molecular docking showed a good score of 1276.5 and molecular dynamics simulations confirmed the construct's efficacy, demonstrating its promise as a good candidate for listeriosis prophylaxis. The population coverage was as high as 91.04% with a good immune response, indicating good antigen presentation with dendritic cells and production of memory cells. CONCLUSIONS: The findings of this study highlight the potential of the designed chimeric peptide construct as an effective subunit vaccine against Listeria, paving the way for future advances in preventive methods and vaccine design.

A Reliable Multifaceted Solution against Foodborne Viral Infections: The Case of RiLK1 Decapeptide.

Food-borne transmission is a recognized route for many viruses associated with gastrointestinal, hepatic, or neurological diseases. Therefore, it is essential to identify new bioactive compounds with broad-spectrum antiviral activity to exploit innovative solutions against these hazards. Recently, antimicrobial peptides (AMPs) have been recognized as promising antiviral agents. Indeed, while the antibacterial and antifungal effects of these molecules have been widely reported, their use as potential antiviral agents has not yet been fully investigated. Herein, the antiviral activity of previously identified or newly designed AMPs was evaluated against the non-enveloped RNA viruses, hepatitis A virus (HAV) and murine norovirus (MNV), a surrogate for human norovirus. Moreover, specific assays were performed to recognize at which stage of the viral infection cycle the peptides could function. The results showed that almost all peptides displayed virucidal effects, with about 90% of infectivity reduction in HAV or MNV. However, the decapeptide RiLK1 demonstrated, together with its antibacterial and antifungal properties, a notable reduction in viral infection for both HAV and MNV, possibly through direct interaction with viral particles causing their damage or hindering the recognition of cellular receptors. Hence, RiLK1 could represent a versatile antimicrobial agent effective against various foodborne pathogens including viruses, bacteria, and fungi.

Assessment of the Microbiological Quality and Effect of Public Health of Ready-to-Eat Salad Samples in Isparta.

Salmonella spp. and Citrobacter spp. are among the microorganisms causing important foodborne outbreaks. In this study, it was tried to determine the presence and rate of Salmonella spp. and Citrobacter spp. in salad samples collected from certain regions of province of Isparta in Türkiye. A total of 50 salad samples were analyzed. Classical culture technique was used for microbiological analysis of salad samples. Suspected isolates obtained were identified using the VITEK-2 system. Although no negative visual changes were observed in the salad samples used in the study, it was determined that the number of Gram-negative microorganisms was very high and six salad samples were not suitable for public health. In 50 salad samples, 2% Salmonella and 4% Citrobacter freundii were detected. In addition, it was determined that the Salmonella strain isolated from the salad sample was resistant to three different antibiotics and Citrobacter was resistant to two different antibiotics. Salmonella spp. and Citrobacter spp. are considered very dangerous to public health because they are associated with foodborne outbreaks and can develop antibiotic resistance very quickly. Salad producers should try to reduce the possibility of microbial contamination by using different technologies.

Characterization of Antimicrobial Activities of Bifidobacterium lactis BB-12 and Their Inhibitory Effect Against Some Foodborne Pathogens.

Bifidobacterium animalis subsp. lactis BB-12, a probiotic, has shown potential to promote health benefits and control pathogens. This study aimed to investigate the effectiveness of BB-12 and its cell-free supernatant (CFS) in inhibiting the growth of Listeria monocytogenes and Salmonella enterica serovar Typhimurium. To assess the antimicrobial activity of BB-12, agar well diffusion, disk diffusion, and minimum inhibitory concentration (MIC) tests were conducted. The bicinchoninic acid (BCA) assay was performed to measure the protein concentration in CFS. The study's results indicated that the BB-12 strain inhibited the pathogens' growth. The disk diffusion test using BB-12 showed inhibitory results ranging from 11 to 14 mm for both bacteria. The agar well diffusion test reported the zone of inhibition ranging from 11.6 to 16 mm for both bacteria. The MIC test was conducted as a confirmatory test, which demonstrated the highest inhibitory zone using 2 McFarland (6 × 108 CFU/mL) concentrations of probiotics on L. monocytogenes (44.98%) and S. Typhimurium (66.41%). The disk diffusion test revealed that the probiotic CFS had a significant inhibitory impact on S. Typhimurium with a 16.6 mm zone of inhibition. The BCA test findings indicated that the 24- and 48-h CFSs exhibited inhibitory properties against infections. Notably, the 24-h CFS, including a protein level of 78.47 µg/mL, demonstrated a more pronounced inhibitory impact on both pathogens. The findings highlight that utilizing the BB-12 strain and its CFS can serve as a viable approach to battle infections, enhancing food safety and public health.

Antibacterial activity of flavonoid extracts from Enteromorpha intestinalis and Caulerpa prolifera against multidrug-resistant foodborne bacterial isolates.

Background: Food poisoning caused by bacterial agents is a worldwide problem, usually accompanied by unpleasant symptoms and may be severe leading to death. Natural compounds from marine algae namely flavonoids may play a role in the remedy of this condition. Aim: This research aims to assess the potency of flavonoids extracted from Enteromorpha intestinalis and Caulerpa prolifera as antibacterial agents. Methods: Enteromorpha intestinalis was collected from Western Libyan Coast and C. prolifera was collected from Farwa Island. The antimicrobial activity and determination of minimum inhibitory concentration of algal flavonoid-containing extracts was performed in vitro against some positive and negative Gram bacteria. Results: Crude extract containing flavonoids from E. intestinalis was more effective than C. prolifera extract against Staphylococcus aureus with antimicrobial essay (25-28 + 1 and 14.5-37.5 + 0.5-1.5), MIC (50 and 50-250 µg/ml), MBC (75 and 75-250 µg/ml). In Bacillus cereus, the antimicrobial assay (19-24.5 + 0.5-1.5: 24 + 1), MIC (50-250 + 100 µg/ml), and MBC (250 and 125 µg/ml). On the other hand, flavonoids containing extract from C. prolifera were more effective than E. intestinalis against Enterohemorrhagic Escherichia coli O157 EHEC O157 (25-28 + 1: 14-18.5 + 0.5-1.5), MIC (100-250:100-500 µg/ml), and MBC (150-250 and 250-500 µg/ml). Salmonella enterica qualitatively combat by flavonoid from E. intestinalis (13.5-14 + 0.5-1: 10.5-13.5 + 0.5-1.5), MIC (100-250: 250 µg/ml), and MBC (100-250: 250 µg/ml). Flavonoids from C. prolifera (4 strains: 2 strains) were effective against S. enterica. Crude flavonoids from both algae were not effective against Bacillus pumilus. Conclusion: Data from this study could conclude that flavonoid extracts from E. intestinalis and C. prolifera could be used against foodborne bacterial agents.

Nanomaterials with Enzyme-like Properties for Combatting Foodborne Pathogen Infections: Classifications, Mechanisms, and Applications in Food Preservation.

During the transportation and storage of food, foodborne spoilage caused by bacterial and biofilm infection is prone to occur, leading to issues such as short shelf life, economic loss, and sensory quality instability. Therefore, the development of novel and efficient antibacterial agents capable of efficiently inhibiting bacteria throughout various stages of food processing, transportation, and storage is strongly recommended by researchers. The emergence of nanozymes is considered to be an effective candidate for inhibiting foodborne bacteria agents in the food industry. As potent antibacterial agents, nanozymes have the advantages of low cost, high stability, strong broad-spectrum antibacterial ability, and biocompatibility. Herein, we aim to summarize the classification status of various nanozymes. Furthermore, the general catalytic bacteriostatic mechanism of nanozymes against intracellular bacteria, planktonic bacteria, and biofilm activities are highlighted, mainly concerning the destruction of cell walls and/or membranes, reactive oxygen species regulation, HOBr/Cl generation, damage of intracellular components, and so forth. In particular, the review focuses on the pivotal role of nanozymes as antibacterial agents and delivery vehicles in the fields of food preservation applications. We look forward to the future prospects, especially in the field of food preservation, to promote broader applications based on antimicrobial nanozymes.

A One-Step RPA-CRISPR Assay Using crRNA Based on Suboptimal Protospacer Adjacent Motif for Vibrio vulnificus Detection.

Vibrio vulnificus is a hazardous foodborne pathogen responsible for approximately 95% of seafood-related deaths. This highlights the urgent requirement for specialized detection tools to be developed and used by food enterprises and food safety authorities. The DETECTR (DNA endonuclease targeted CRISPR trans reporter) system that combines CRISPR/Cas and recombinase polymerase amplification (RPA) has been utilized to develop a molecular detection assay for V. vulnificus. However, because the incompatibility between RPA and Cas12a cleavage has not been addressed, it is a two-step assay that lacks convenience and presents contamination risk. Here, we developed a one-step RPA-CRISPR assay for V. vulnificus using a special crRNA targeting a sequence with a suboptimal protospacer adjacent motif (PAM). The entire assay, conducted at 37°C, takes only 40-60 min, yields results visualized under blue light, and exhibits exceptional specificity and sensitivity (detecting 4 pathogen genome copies per reaction). This study offers a valuable tool for detecting V. vulnificus, aiding in foodborne infection prevention, and exemplifies one-step RPA-CRISPR assays managing Cas-cleavage activity through PAM adjustments.

The Knowledge and Practices Toward Food Safety Measures at Home in the Lebanese Community.

Hygienic measures practiced at home are highly related to the occurrence of food-borne diseases during food production, storage, and handling. Contaminated food remains a major cause of several diarrheal diseases, hospitalizations, and spikes in medical expenses. In our current study, we aimed to assess the knowledge of food safety and the food safety and hygiene practices at home among the Lebanese population. A cross-sectional study was conducted through an online questionnaire including two sections. The first section included socio-demographic characteristics of participants, whereas the second section included questions related to practices and knowledge about food safety, divided into five parts; personal hygiene practices, dry and cold storage, sanitizing and cleaning and food intoxication. A total of 1101 Lebanese above 18 years participated and provided their responses to the questionnaire. Overall, the majority of participants had fair knowledge about food safety where 96.8% of the participants answered correctly about preventing microbial growth on food. 77.9% of those participants acquired their knowledge about food safety from articles, workshops, or the internet. Moreover, females, people with children and those who cook for themselves scored significantly higher than others (68.8, 70.6, and 70%, respectively). In comparison to younger participants (67.8%), older participants (50+ and 30-49) scored higher at 69.7% and 68.9%, respectively. Higher scores were obtained for questions related to storing dried foods/meat and poultry products with percentages 91.4 and 87.8%, respectively. However, lower scores were noticed on questions related to washing raw chicken before handling and storing eggs (9.7 and 12.3%, respectively). Altogether, our results revealed the need for directed food safety awareness campaigns at the national level to educate the Lebanese community about domestic food handling practices. We believe these campaigns can significantly reduce related diseases and hospitalizations.

Impact of Foodborne Disease in Taiwan during the COVID-19 Pandemic.

Background and Objectives: The coronavirus disease 2019 (COVID-19) pandemic originated in Wuhan, China, in December 2019, the first case diagnosed since January 2020 in Taiwan. The study about the potential impact of the COVID-19 pandemic on event, location, food source, and pathogens of foodborne disease (FBD) is limited in Taiwan. Our aim in this study is to investigate FBD in the context of the COVID-19 pandemic. Materials and Methods: We collected publicly available annual summary data from the FBD dataset in the Taiwan Food and Drug Administration and Certifiable Disease on reported FBD in Taiwan from 2019 to 2020. We used logistic regression to evaluate changes in the occurrence or likelihood of FBD cases and Poisson regression to examine the relative risk (RR) between FBD and climate factors. Results: Similar events occurred in 2019 and 2020, but the total number of FBD cases decreased from 6935 in 2019 to 4920 in 2020. The places where FBD decreased were in schools, hospitals, outdoors, vendors, and exteriors. The top place in FBD shifted from schools to restaurants. The top food source for FBD has changed from boxed food to compound food. Bacillus cereus and Salmonella emerged as the top two observed bacterial pathogens causing FBD. The risk of FBD cases increased with a higher air temperature, with an RR of 1.055 (1.05-1.061, p < 0.001) every 1 °C. Conclusion: The incidence of FBD decreased significantly during the COVID-19 pandemic in Taiwan. This decline may be attributed to protective measures implemented to control the spread of the virus. This shift in locations could be influenced by changes in public behavior, regulations, or other external factors. The study emphasizes the importance of understanding the sources and effectiveness of severe infection prevention policies. The government can use these findings to formulate evidence-based policies aimed at reducing FBD cases and promoting public health. Consumers can reduce the risk of FBD by following safe food handling and preparation recommendations.

Systematic review on microbiome-related nutritional interventions interfering with the colonization of foodborne pathogens in broiler gut to prevent contamination of poultry meat.

This systematic review aimed to compile the available body of knowledge about microbiome-related nutritional interventions contributing to improve the chicken health and having an impact on the reduction of colonization by foodborne pathogens in the gut. Original research articles published between 2012 and 2022 were systematically searched in Scopus and PubMed. A total of 1,948 articles were retrieved and 140 fulfilled the inclusion criteria. Overall, 73 papers described 99 interventions against colonization by Escherichia coli and related organisms; 10 papers described 15 interventions against Campylobacter spp.; 36 papers described 54 interventions against Salmonella; 40 papers described 54 interventions against Clostridium perfringens. A total of 197 microbiome-related interventions were identified as effective against one or more of the listed pathogens and included probiotics (n = 80), prebiotics (n = 23), phytobiotics (n = 25), synbiotics (n = 12), organic acids (n = 12), enzymes (n = 4), essential oils (n = 14) and combination of these (n = 27). The identified interventions were mostly administered in the feed (173/197) or through oral gavage (11/197), in the drinking water (7/197), in ovo (2/197), intra amniotic (2/197), in fresh or reused litter (1/197) or both in the feed and water (1/197). The interventions enhanced the beneficial microbial communities in the broiler gut as Lactic acid bacteria, mostly Lactobacillus spp., or modulated multiple microbial populations. The mechanisms promoting the fighting against colonization by foodborne pathogens included competitive exclusion, production of short chain fatty acids, decrease of gut pH, restoration of the microbiome after dysbiosis events, promotion of a more stable microbial ecology, expression of genes improving the integrity of intestinal mucosa, enhancing of mucin production and improvement of host immune response. All the studies extracted from the literature described in vivo trials but performed on a limited number of animals under experimental settings. Moreover, they detailed the effect of the intervention on the chicken gut without details on further impact on poultry meat safety.