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This study examined the impact of the 2017 Veterinary Feed Directive (VFD) rule changes on the prevalence of tetracycline-resistant and erythromycin-resistant bacteria (Salmonella spp., Campylobacter spp., and Escherichia coli) in cecal samples of food animals (cattle, swine, chicken, and turkey) at US slaughterhouses. Multivariable logistic regression was used to analyze 2013–2019 cecal samples of food-producing animals surveillance data from the National Antimicrobial Resistance Monitoring System (NARMS) in the U.S. The variables included year (used to evaluate VFD rule changes), host, and quarter of year. The analysis of surveillance data showed that the VFD rule changes have varying effects on tetracycline-resistant and erythromycin-resistant bacteria in food animals. For example, the odds of detecting tetracycline-resistant Salmonella spp. decreased in cattle but increased in chickens following the implementation of the VFD rule changes. Similarly, the odds of detecting tetracycline-resistant Escherichia coli decreased in chickens but increased in swine after the VFD rule changes. The odds of detecting erythromycin-resistant Campylobacter spp. increased in cattle but decreased in chickens after the VFD rule changes. In conclusion, the implementation of VFD rule changes has been beneficial in reducing the odds of detecting tetracycline-resistant Escherichia coli and erythromycin-resistant Campylobacter spp. in chickens, as well as tetracycline-resistant Salmonella spp. in cattle at US slaughterhouses. [ABSTRACT FROM AUTHOR]

In March 2024, clade 2.3.4.4b H5N1 highly pathogenic avian influenza virus (HPAIV) was detected in dairy cattle in the US, and it was discovered that the virus could be detected in raw milk. Although affected cow’s milk is diverted from human consumption and current pasteurization requirements are expected to reduce or eliminate infectious HPAIV from the milk supply, a study was conducted to characterize whether the virus could be detected by quantitative real-time RT-PCR (qrRT-PCR) in pasteurized retail dairy products and, if detected, to determine whether the virus was viable. From 18 April to 22 April 2024, a total of 297 samples of Grade A pasteurized retail milk products (23 product types) were collected from 17 US states that represented products from 132 processors in 38 states. Viral RNA was detected in 60 samples (20.2%), with qrRT-PCRbased quantity estimates (non-infectious) of up to 5.4log1050% egg infectious doses per mL, with a mean and median of 3.0log10/mL and 2.9log10/mL, respectively. Samples that were positive for type A influenza by qrRT-PCR were confirmed to be clade 2.3.4.4 H5 HPAIV by qrRT-PCR. No infectious virus was detected in any of the qrRT-PCR-positive samples in embryonating chicken eggs. Further studies are needed to monitor the milk supply, but these results provide evidence that the infectious virus did not enter the US pasteurized milk supply before control measures for HPAIV were implemented in dairy cattle. IMPORTANCE Highly pathogenic avian influenza virus (HPAIV) infections in US dairy cattle were first confirmed in March 2024. Because the virus could be detected in raw milk, a study was conducted to determine whether it had entered the retail food supply. Pasteurized dairy products were collected from 17 states in April 2024. Viral RNA was detected in one in five samples, but infectious virus was not detected. This provides a snapshot of HPAIV in milk products early in the event and reinforces that with current safety measures, infectious viruses in milk are unlikely to enter the food supply. [ABSTRACT FROM AUTHOR]

Abstract Recently, the use of D‐amino acids as food preservatives has attracted considerable attention because these natural compounds do not have adverse effects on human health. In addition, D‐amino acids such as D‐tryptophan can reduce the harmful effects of other treatments. For instance, the use of D‐tryptophan in food reduces the requirement for high temperatures and their damaging effects on nutrients such as proteins and vitamins. The purpose of this systematic review was to investigate the antimicrobial effect of D‐tryptophan on food‐borne pathogens in vitro and in food models. To identify related studies, scientific digital databases such as PubMed, Science Direct, and Google Scholar were searched from January 2000 to February 2023. The results of the studies showed that when D‐tryptophan was used with other stresses such as using different salt concentrations, refrigeration, or high temperatures, it showed significant antimicrobial effects on Gram‐positive and Gram‐negative food‐borne pathogens, and antibiofilm impacts were also observed with D‐tryptophan. Since studies have shown that the antimicrobial activity of D‐tryptophan depends on several factors, including the pathogen strain, the type of stress, and the concentration of D‐tryptophan, and every article has focused on one of these factors, there is a need for a systematic review that summarizes and concludes the effect of all these factors on the antimicrobial activity of D‐tryptophan against food‐borne pathogens.

Recently, the use of D‐amino acids as food preservatives has attracted considerable attention because these natural compounds do not have adverse effects on human health. In addition, D‐amino acids such as D‐tryptophan can reduce the harmful effects of other treatments. For instance, the use of D‐tryptophan in food reduces the requirement for high temperatures and their damaging effects on nutrients such as proteins and vitamins. The purpose of this systematic review was to investigate the antimicrobial effect of D‐tryptophan on food‐borne pathogens in vitro and in food models. To identify related studies, scientific digital databases such as PubMed, Science Direct, and Google Scholar were searched from January 2000 to February 2023. The results of the studies showed that when D‐tryptophan was used with other stresses such as using different salt concentrations, refrigeration, or high temperatures, it showed significant antimicrobial effects on Gram‐positive and Gram‐negative food‐borne pathogens, and antibiofilm impacts were also observed with D‐tryptophan. Since studies have shown that the antimicrobial activity of D‐tryptophan depends on several factors, including the pathogen strain, the type of stress, and the concentration of D‐tryptophan, and every article has focused on one of these factors, there is a need for a systematic review that summarizes and concludes the effect of all these factors on the antimicrobial activity of D‐tryptophan against food‐borne pathogens. [ABSTRACT FROM AUTHOR]

The extensive use of synthetic antibiotics and insecticides in controlling microbes and insect spreads led to a build-up of resistance strains and caused negative impacts on human health through bioaccumulation in food and the environment. The present study assessed the antimicrobial activity of Amphora coffeaeformis (A. coffeaeformis) ethanolic extract against foodborne pathogenic microbes as well as assessed its ability to control Culex pipiens L. (C. pipiens) larvae. The gas chromatography-mass spectrum (GC/MS) and high-performance liquid chromatography (HPLC) analysis showed that A. coffeaeformis extract was rich with phenolic compounds, organosulfur compounds, carboxylic acids, amino acids, organic quinoline, dipeptide and monosaccharide. The extract of A. coffeaeformis showed antibacterial capability against all tested bacteria, with minimal inhibition concentration (MIC) values ranging from 20.2 to 66.7 g/mL, as well as antifungal effects against all tested mycotoxigenic fungi, with MIC values ranging from 113.62, 68.95 and 49.37 µg/mL mg/mL. Furthermore, the extract showed high larvicidal activity against C. pipiens larvae at 24, 48, and 72 h, respectively. LC50 values decreased gradually with the treatment period. Likewise, the adult emergence and growth index were significantly negatively related to increasing extract concentrations. Moreover, the extract exhibited a noteworthy depletion in the reproductive potential and increased sterility index of C. pipiens females developed of treated larvae. Besides a reduction in egg hatching percent, increase pupal malformations, increase and adult abnormalities. C. pipiens biochemical markers; glutathione-S-transferase, carboxylesterase, acetylcholinesterase, a-esterase and cytochrome P-450 showed significant alteration after the extract exposure. Overall, these findings promise the application possibility of A. coffeaeformis extract as a biopreservator against foodborne pathogens and as bioinsecticides for mosquito control as well. [ABSTRACT FROM AUTHOR]

The presence of microbial pathogens in ready-to-eat produce represents a serious health problem. The antibacterial activity of cinnamon (Cinnamomum zeylanicum) and clove (Syzygium aromaticum L. Merr. & Perry) essential oils (EOs) was determined toward food-borne pathogens by agar disk diffusion and minimum inhibitory concentration (MIC) assays. The growth kinetics of all strains, both in a buffer suspension assay and "on food" in artificially contaminated samples, were also investigated. The two EOs demonstrated a good antibacterial effect both alone and in combination (EO/EO). The use of EO/EO led to a synergistic antibacterial effect, also confirmed by the growth kinetics studies, where the EOs were active after 10 h of incubation (p < 0.0001) at significantly lower concentrations than those when alone. In the "on food" studies performed on artificially contaminated fruit samples stored at 4 °C for 8 days, the greatest killing activity was observed at the end of the trial (8 days) with a reduction of up to 7 log CFU/g compared to the control. These results confirm the good antibacterial activity of the EOs, which were more effective when used in combination. Data from the "on food" studies suggest cinnamon and clove essential oils, traditionally used in the food industry, as a possible natural alternative to chemical additives. [ABSTRACT FROM AUTHOR]

Cryptophytes are a promising source of bioactive compounds that have not been fully explored. This research investigated the antimicrobial activity of total phenolic compounds (TPC) and exopolysaccharides (EPS) extracted from several cryptophytes against a range of harmful foodborne bacteria and fungi. To measure the minimum inhibitory concentration (MIC) value, the broth microdilution method was used. In the antibacterial evaluation of TPC, the MIC ranged between 31.25 and 500 μg/mL, while for the antifungal activity test, it varied from 31.25 to 125 μg/mL. In the antibacterial activity test of EPS, the MIC values ranged from 125 to 1,000 μg/mL, whereas in the antifungal susceptibility test, it ranged between 62.5 and 1,000 μg/mL. The most resistant pathogen against TPC was Escherichia coli, while Campylobacter jejuni was the most susceptible. In the case of EPS, the most resistant pathogen was Salmonella Typhimurium, while Aspergillus versicolor exhibited the highest susceptibility. Overall, in terms of antimicrobial activity, TPC was more effective than EPS. Finally, the tolerance level (TL) for TPC and EPS was ≤4 in all tested samples, indicating their bactericidal/fungicidal mechanism of action. In conclusion, TPC and EPS isolated from cryptophytes demonstrated remarkable antimicrobial properties and ability to fully eradicate pathogens, and could be considered as natural preservatives in the food industry.

In the food industry, successful bacterial pathogen colonization and persistence begin with their adhesion to a surface, followed by the spatial development of mature biofilm of public health concerns. Compromising bacterial settlement with natural inhibitors is a promising alternative to conventional anti-fouling treatments typically based on chemical biocides that contribute to the growing burden of antimicrobial resistance. In this study, three extracellular polymeric substance (EPS) fractions extracted from microalgae biofilms of Cylindrotheca closterium (fraction C) and Tetraselmis suecica (fraction Ta rich in insoluble scale structure and fraction Tb rich in soluble EPS) were screened for their anti-adhesive properties, against eight human food-borne pathogens belonging to Escherichia coli, Staphylococcus aureus, Salmonella enterica subsp. enterica, and Listeria monocytogenes species. The results showed that the fraction Ta was the most effective inducing statistically significant reduction for three strains of E. coli, S. aureus, and L. monocytogenes. Overall, EPSs coating on polystyrene surfaces of the different fractions increased the hydrophilic character of the support. Differences in bacterial adhesion on the different coated surfaces could be explained by several dissimilarities in the structural and physicochemical EPS compositions, according to HPLC and ATR-FTIR analysis. Interestingly, while fractions Ta and Tb were extracted from the same microalgal culture, distinct adhesion patterns were observed, highlighting the importance of the extraction process. Overall, the findings showed that EPS extracted from microalgal photosynthetic biofilms can exhibit anti-adhesive effects against food-borne pathogens and could help develop sustainable and non-toxic anti-adhesive surfaces for the food industry. Key points: •EPSs from a biofilm-based culture of C. closterium/T. suecica were characterized. •Microalgal EPS extracted from T. suecica biofilms showed bacterial anti-adhesive effects. •The anti-adhesive effect is strain-specific and affects both Gram − and Gram + bacteria. [ABSTRACT FROM AUTHOR]

The addition of carminative essential oils could be an approach for food preservation and would minimize or substitute chemical preservatives. In the present study, essential oils (n=11) namely, Anethum sowa, Cinnamomum zeylanicum, Citrus bergamia, Cymbopogon flexuosus, Cymbopogon martini, Cymbopogon winterianus, Elettaria cardamomum, Mentha arvensis, Ocimum basilicum, Salvia sclarea and Trachyspermum ammi, were screened against Aeromonas hydrophila and Listeria monocytogenes. The largest diameters of zone of inhibition, 19.9 ± 0.33 mm and 21.7 ± 0.58 mm, were exhibited by T. ammi essential oil against Aeromonas hydrophila and Listeria monocytogenes, respectively. Growth inhibition studies for T. ammi essential oil, sodium nitrite and their combinations were also carried out on cucumber, apple, gram flour soup and mutton broth models. The combination of T. ammi essential oil and sodium nitrite depicted synergism and was also effective in reducing the bacterial counts in artificially inoculated food systems. [ABSTRACT FROM AUTHOR]

Purpose: Nature and occurrence of food-borne pathogens in raw and processed food products evolved greatly in the past few years due to new modes of transmission and resistance build-up against sundry micro-/macro-environmental conditions. Assurance of food health and safety thus gained immense importance, for which bio-sensing technology proved very promising in the detection and quantification of food-borne pathogens. Considering the importance, different studies have been performed, and different biosensors have been developed. This study aims to summarize the different biosensors used for the deduction of food-borne pathogens. Design/methodology/approach: The present review highlights different biosensors developed apropos to food matrices, factors governing their selection, their potential and applicability. The paper discusses some related key challenges and constraints and also focuses on the needs and future research prospects in this field. Findings: The shift in consumers' and industries' perceptions directed the further approach to achieve portable, user and environmental friendly biosensing techniques. Despite of these developments, it was still observed that the comparison among the different biosensors and their categories proved tedious on a single platform; since the food matrices tested, pathogen detected or diagnosed, time of detection, etc., varied greatly and very few products have been commercially launched. Conclusively, a challenge lies in front of food scientists and researchers to maintain pace and develop techniques for efficiently catering to the needs of the food industry. Research limitations/implications: Biosensors deduction limit varied with the food matrix, type of organism, material of biosensors' surface, etc. The food matrix itself consists of complex substances, and various types of food are available in nature. Considering the diversity of food there is a need to develop a universal biosensor that can be used for all the food matrices for a pathogen. Further research is needed to develop a pathogen-specific biosensor that can be used for all the food products that may have accuracy to eliminate the traditional method of deduction. Originality/value: The present paper summarized and categorized the different types of biosensors developed for food-borne pathogens. [ABSTRACT FROM AUTHOR]

The present study aimed to characterize the mode of action of a novel antimicrobial peptide isolated from egg yolk hydrolysate. The EYHp6, KGGDLGLFEPTL, exhibited inhibition against Salmonella enterica serovar Typhimurium TISTR 292 and S. enterica serovar Enteritidis DMST 15679 with a MIC value of 2 mM. In contrast, S. enterica serovar Newport ATCC 6962 and other strains of Typhimurium and Enteritidis were inhibited at 4 mM. EYHp6 increased the cell membrane permeability of S. Typhimurium TISTR 292, leading to DNA leakage. Membrane integrity determined by propidium iodide and SYTO9 staining visualized by confocal microscopy demonstrated that EYHp6 at 1 × MIC induced disruption of cell membranes. Electron microscopy revealed that treatment of S. Typhimurium with EYHp6 led to damage to the cell membrane, causing the leakage of intracellular contents. Synchrotron-based Fourier-transform infrared spectroscopy indicated that EYHp6 killed S. Typhimurium by targeting fatty acids and nucleic acids in the cell membrane. The peptide did not show hemolytic activity up to 4 mM. These findings suggest that EYHp6 could be a promising antibacterial agent for controlling the growth of S. enterica. [ABSTRACT FROM AUTHOR]

The addition of carminative essential oils could be an approach for food preservation and would minimize or substitute chemical preservatives. In the present study, essential oils (n=11) namely, Anethum sowa, Cinnamomum zeylanicum, Citrus bergamia, Cymbopogon flexuosus, Cymbopogon martini, Cymbopogon winterianus, Elettaria cardamomum, Mentha arvensis, Ocimum basilicum, Salvia sclarea and Trachyspermum ammi, were screened against Aeromonas hydrophila and Listeria monocytogenes. The largest diameters of zone of inhibition, 19.9 ± 0.33 mm and 21.7 ± 0.58 mm, were exhibited by T. ammi essential oil against Aeromonas hydrophila and Listeria monocytogenes, respectively. Growth inhibition studies for T. ammi essential oil, sodium nitrite and their combinations were also carried out on cucumber, apple, gram flour soup and mutton broth models. The combination of T. ammi essential oil and sodium nitrite depicted synergism and was also effective in reducing the bacterial counts in artificially inoculated food systems.

Background: Food safety is an important subject that global cheese industry increases the awareness upon. This urges these economic sectors to elevate the level of research to minimize cheese contamination with pathogenic bacteria, such as Salmonella. Aim: Based on these merits, this study was conducted to genotype Salmonella spp isolated from cheese samples of local stores in Al-Diwaniyah City, Iraq. Methods: The study used 41 samples of local fresh unsalted white cheese in a selective-growth based isolation of Salmonella. These isolates were confirmed utilizing a slide-agglutination (SA) test and VITEK® 2 system (V2S). Then, the isolates were subjected to conventional PCR and sequencing techniques that both targeted the 16S rRNA gene. For subtyping, the Salmonella isolates were subjected to a random amplified polymorphic DNA (RAPD)-PCR method. Results: The results of both SA and V2S revealed the presence of 14 (34.2%) isolates of Salmonella spp. in the cheese samples. The PCR confirmed 6 (42.9%) of these isolates, which further were defined with close nucleotide similarity (98.03%) and (97.88%) to different world isolates, such as Salmonella enterica subsp. Arizonae and Salmonella enterica subsp. enterica serovar Typhi, respectively. The RAPD-PCR findings showed different fragments for all the tested isolates. Conclusion: The present study indicates that the samples of the local fresh unsalted white cheese contain different Salmonella genotypes, which could be originated from different contamination sources. [Open Vet J 2023; 13(10.000): 1277-1282]

This study examined the impact of the 2017 Veterinary Feed Directive (VFD) rule changes on the prevalence of tetracycline-resistant and erythromycin-resistant bacteria (Salmonella spp., Campylobacter spp., and Escherichia coli) in cecal samples of food animals (cattle, swine, chicken, and turkey) at US slaughterhouses. Multivariable logistic regression was used to analyze 2013–2019 cecal samples of food-producing animals surveillance data from the National Antimicrobial Resistance Monitoring System (NARMS) in the U.S. The variables included year (used to evaluate VFD rule changes), host, and quarter of year. The analysis of surveillance data showed that the VFD rule changes have varying effects on tetracycline-resistant and erythromycin-resistant bacteria in food animals. For example, the odds of detecting tetracycline-resistant Salmonella spp. decreased in cattle but increased in chickens following the implementation of the VFD rule changes. Similarly, the odds of detecting tetracycline-resistant Escherichia coli decreased in chickens but increased in swine after the VFD rule changes. The odds of detecting erythromycin-resistant Campylobacter spp. increased in cattle but decreased in chickens after the VFD rule changes. In conclusion, the implementation of VFD rule changes has been beneficial in reducing the odds of detecting tetracycline-resistant Escherichia coli and erythromycin-resistant Campylobacter spp. in chickens, as well as tetracycline-resistant Salmonella spp. in cattle at US slaughterhouses.

Bangladesh's commercial poultry production is growing rapidly, including the commercial processing of poultry. This expansion of poultry processing plants is fueled by the belief that this sub-sector provides safer food and has less food-borne disease risks compared to traditional live bird markets (LBMs). The purpose of this study is to describe Bangladesh's dressed and processed poultry production and distribution network (PDN), identify what and where quality control occurs, and suggest where improvements could be made. Engaging with PDN for dressed and processed poultry, we used in-depth interviews with key informants to identify the stakeholders involved and their connections with other poultry PDNs. In addition, we mapped out the supply and distribution of dressed and processed poultry and quality control processes occurring throughout the network. We argue that dressed and processed poultry PDNs are closely connected with traditional PDNs such as LBMs, with multiple crossover points between them. Also, there is a lack of consistency in quality control testing and a lack of meat traceability. Consequently, perceptions of dressed and processed poultry being safer than birds from LBMs needs to be treated with caution. Otherwise, unsubstantiated consumer confidence in dressed poultry may inadvertently increase the risk of food-borne diseases from these products. [ABSTRACT FROM AUTHOR]

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ABSTRACTThe resolution of variation within species is critical for interpreting and acting on many microbial measurements. In the key foodborne pathogens Salmonella and Escherichia coli, the primary subspecies classification scheme used is serotyping: differentiating variants within these species by surface antigen profiles. Serotype prediction from whole-genome sequencing (WGS) of isolates is now seen as comparable or preferable to traditional laboratory methods where WGS is available. However, laboratory and WGS methods depend on an isolation step that is time-consuming and incompletely represents the sample when multiple strains are present. Community sequencing approaches that skip the isolation step are, therefore, of interest for pathogen surveillance. Here, we evaluated the viability of amplicon sequencing of the full-length 16S rRNA gene for serotyping Salmonella enterica and E. coli. We developed a novel algorithm for serotype prediction, implemented as an R package (Seroplacer), which takes as input full-length 16S rRNA gene sequences and outputs serovar predictions after phylogenetic placement into a reference phylogeny. We achieved over 89% accuracy in predicting Salmonella serotypes on in silico test data and identified key pathogenic serovars of Salmonella and E. coli in isolate and environmental test samples. Although serotype prediction from 16S rRNA gene sequences is not as accurate as serotype prediction from WGS of isolates, the potential to identify dangerous serovars directly from amplicon sequencing of environmental samples is intriguing for pathogen surveillance. The capabilities developed here are also broadly relevant to other applications where intraspecies variation and direct sequencing from environmental samples could be valuable.IMPORTANCEIn order to prevent and stop outbreaks of foodborne pathogens, it is important that we can detect when pathogenic bacteria are present in a food or food-associated site and identify connections between specific pathogenic bacteria present in different samples. In this work, we develop a new computational technology that allows the important foodborne pathogens Escherichia coli and Salmonella enterica to be serotyped (a subspecies level classification) from sequencing of a single-marker gene, and the 16S rRNA gene often used to surveil bacterial communities. Our results suggest current limitations to serotyping from 16S rRNA gene sequencing alone but set the stage for further progress that we consider likely given the rapid advance in the long-read sequencing technologies and genomic databases our work leverages. If this research direction succeeds, it could enable better detection of foodborne pathogens before they reach the public and speed the resolution of foodborne pathogen outbreaks.

Food-borne pathogens pose significant risks to public health and food safety, necessitating the development of rapid, sensitive, and multiplex detection techniques. Biosensor is one of the important tool to detect the analyte with high sensitivity. Now a days multiplex based approaches were in high demands due to its various advantages. One of the important advantages of multiplex aptasensor is it can detects various food borne pathogens on a single platform which also reduced the coast of detection and shows high selectivity due to integration of multiplex platform with aptamer. The emergence of aptamers as versatile recognition elements is discussed, emphasizing their advantages over antibodies in terms of stability, cost-effectiveness, and ease of synthesis. A critical analysis of state-of-the-art multiplex aptasensor platforms is then presented. These platforms encompass a wide range of detection modalities, such as electrochemical, optical, and microfluidic systems. Moreover, the review explores recent examples of multiplex aptasensor applications for detecting a variety of Food-borne pathogens, including bacteria (e.g., Salmonella, Escherichia coli, Listeria) and viruses (e.g., Norovirus, Hepatitis A). The discussion encompasses both in vitro and on-site detection scenarios, demonstrating the versatility and potential impact of multiplex aptasensors across different stages of the food supply chain. The paper concludes by highlighting the transformative potential of multiplex aptasensor detection techniques in enhancing food safety, public health, and surveillance strategies.

This study aimed to genotype isolates of Toxoplasma gondii obtained from samples of brain, diaphragm and heart of goats and sheep intended for human consumption in the State of Paraíba, Brazil. Tissue samples from 14 animals, goats (n = 5) and lambs (n = 9), were sourced from public slaughterhouses in seven cities and bio-assayed in mice. The brains of the mice were utilized for DNA extraction. Genotyping was carried out by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) using 10 markers (SAG1, SAG2, SAG3, BTUB, c22-8, PK1, GRA6, L358, c-29-2 and Apico). A total of 10 isolates were fully genotyped (i.e. at all loci), three from goats and seven from sheep, revealing five distinct genotypes: #13 (n = 4); #48 (n = 3); #57 (n = 1); #273 (n = 1); and one new genotype that had not been previously described. Genotype #13 is frequently found in the Northeast of Brazil and represents a clonal lineage circulating in this region and was the most prevalent genotype identified (n = 4). Moreover, in the present study genotypes #13, #48, #57, and #273 were documented for the first time in sheep from Brazil, and the novel genotype was isolated from a goat. Our findings align with previous studies on T. gondii from Brazil, where new genotypes are continuously being identified, highlighting a high level of genetic diversity of T. gondii isolates in the country.

ABSTRACT Microorganisms frequently colonize surfaces within food production facilities. Detection of Listeria monocytogenes in this setting relies on culture-dependent methods, but the complex dynamics of bacterial interactions within these environments and their impact on pathogen detection remain largely unexplored. To address this challenge, we applied both 16S rRNA and shotgun quasimetagenomic (enriched microbiome) sequencing of swab culture enrichments from five seafood and seven dairy production environments. Utilizing 16S rRNA amplicon sequencing, we observed variability between 355 samples taken from these 12 production facilities and a distinctive microbiome for each environment. With shotgun quasimetagenomic sequencing, we were able to assemble L. monocytogenes metagenome-assembled genomes (MAGs) from 28 of the 32 culture-positive samples. We compared these MAGs to their corresponding whole-genome sequencing assemblies, which resulted in two polyphyletic clades consisting of L. monocytogenes lineages I and II with 13,195 and 25,556 single-nucleotide polymorphism sites, respectively. The remaining four MAGs did not produce sufficient genome coverage. To understand and establish limits for pathogen detection and subtyping using shotgun quasimetagenomics, these same data sets were downsampled in slilico to produce a titration series of abundances of L. monocytogenes and analyzed. Pathogen detection was achieved for all downsampled data sets, even those with only 3× genome coverage. This study contributes to the understanding of microbial diversity within food production environments and presents insights into the level of genome coverage needed in a metagenome sequencing data set to detect, subtype, and source track a foodborne pathogen IMPORTANCE In developed countries, the human diet is predominated by food commodities, which have been manufactured, processed, and stored in a food production facility. Little is known about the application of metagenomic sequencing approaches for detecting foodborne pathogens, such as L. monocytogenes, and characterizing microbial diversity in food production ecosystems. In this work, we investigated the utility of 16S rRNA amplicon and quasimetagenomic sequencing for the taxonomic and phylogenetic classification of Listeria culture enrichments of environmental swabs collected from dairy and seafood production facilities. We demonstrated that single-nucleotide polymorphism (SNP) analyses of L. monocytogenes metagenome-assembled genomes (MAGs) from quasimetagenomic data sets can achieve similar resolution as culture isolate whole-genome sequencing. To further understand the impact of genome coverage on MAG SNP cluster resolution, an in silico downsampling approach was employed to reduce the percentage of target pathogen sequence reads, providing an initial estimate of required MAG coverage for subtyping resolution of L. monocytogenes.

Pseudomonas aeruginosa is a Gram-negative bacillus associated with waterborne diseases. The objective of this study was to determine whether particular P. aeruginosa sequence types (STs) were associated with drinking water contamination in Brazil. This was achieved by searching the Pseudomonas PubMLST database, which contains the records for 8358 strains collected between 1938 and 2023. The majority (97.2%) had the complete 7-loci multilocus sequence typing profile and were assigned to 3486 STs. After eBURST (an algorithm used to infer patterns of evolutionary descent among clusters), 1219 groups with single-locus variant and 575 groups with double-locus variant were formed. Brazil was the South American country with the most isolates (n  = 219, 58.24%), and the Simpson's index was 0.9392. Of the 219 Brazilian isolates, eight were isolated in water and identified as STs 252, 1417, 1605, 2502, 2620, 3078, and 3312. ST252, 1417, and 3078 have already been isolated from clinical cases worldwide. Furthermore, ST1605 and 2620, after the eBURST, they were grouped in the same clonal complex as STs involved in human infections. In conclusion, P. aeruginosa STs involved in human infections were found in bottled drinking water commercialized in Brazil, revealing that these types of drinking waters can be a vehicle of contamination. [ABSTRACT FROM AUTHOR]

The presence of microbial pathogens in ready-to-eat produce represents a serious health problem. The antibacterial activity of cinnamon (Cinnamomum zeylanicum) and clove (Syzygium aromaticum L. Merr. & Perry) essential oils (EOs) was determined toward food-borne pathogens by agar disk diffusion and minimum inhibitory concentration (MIC) assays. The growth kinetics of all strains, both in a buffer suspension assay and “on food” in artificially contaminated samples, were also investigated. The two EOs demonstrated a good antibacterial effect both alone and in combination (EO/EO). The use of EO/EO led to a synergistic antibacterial effect, also confirmed by the growth kinetics studies, where the EOs were active after 10 h of incubation (p < 0.0001) at significantly lower concentrations than those when alone. In the “on food” studies performed on artificially contaminated fruit samples stored at 4 °C for 8 days, the greatest killing activity was observed at the end of the trial (8 days) with a reduction of up to 7 log CFU/g compared to the control. These results confirm the good antibacterial activity of the EOs, which were more effective when used in combination. Data from the "on food" studies suggest cinnamon and clove essential oils, traditionally used in the food industry, as a possible natural alternative to chemical additives.

Although their nutritive value and popularity, milk and dairy products frequently serve as a vehicle for the various pathogens of both human and animal origins. A wide variety of artisanal dairy products are produced in rural areas and villages of Egypt and due to lack of strict hygienic measures and lack of thermal treatment, these products usually harbor a variety of spoilage and possibly food-poisoning microorganisms. The current study aimed to investigate the contamination incidence of raw milk, Kariesh cheese and artisanal Yoghurt (Zabady) by staphylococci, E. coli and Bacillus cereus. Overall, 75 samples of raw milk (100.00%) were contaminated by staphylococci, 54 samples of karish cheese (72.00 %) contained E. coli, meanwhile, 45 samples of Zabady (60.00 %) yielded Bacillus cereus. Additionally, 29 out of 34 E. coli strains recovered from examined samples, were successfully serotyped with correspondence to 9 different serogroups, meanwhile 43 strains from different samples were untypable by available antisera. 80 out of all 162 E. coli isolates (49.38 %) carried haemolytic activity feature, which reflect a great threat towards consumers. Furthermore, the phenotypic AR of S. aureus and E. coli isolates was checked against eleven selected antibiotics. A remarkable variation in phenotypic AR was noticed among strains. Our results denote the high incidence of health hazards in raw milk and its products and the existence of AR S. aureus and E. coli strains isolated from milk and dairy products in rural areas, which could cause human illnesses that are difficult to be treated.

Food contaminated with bacterial pathogens is a great threat to human health and food spoilage, having an impact on public health and the food industry. Research in food safety seeks to develop a practical, rapid, and sensitive detection technique for food-borne pathogens. In the past few decades, real-time quantitative polymerase chain reaction (qPCR) has been developed, and multiplex qPCR is a preferred feature. Multiplex qPCR enables the simultaneous amplification of many targets of interest in one reaction by using more than one pair of primers. In this study, we have developed and evaluated a hydrolysis (TaqMan) probe-based system for simultaneous detection of eight of the most common food-borne pathogens in a single-step procedure by multiplex qPCR. A multicolor combinational probe coding (MCPC) strategy was utilized that allows multiple fluorophores to label different probes in combinatorial manner. This strategy enabled simultaneous detection, identification, and quantification of targeted genes. The efficiency of the individual qPCR reactions for each target gene had values comparable to those established for multiplex qPCR, with detection limits of approximately < 10 copies of DNA per reaction. Pathogen load helps to predict bacteriological quality status in food products and serves to validate the efficiency of procedures to minimize or eliminate their presence, so newly developed multiplex qPCR was quantitative for each pathogen. During sample preparation, a step to concentrate the target organism from a relatively large sample size, remove all potential PCR inhibitors, and yield samples in a volume suitable for qPCR was incorporated.

Toxoplasmosis, caused by the obligate intracellular protozoan Toxoplasma gondii, is a worldwide parasitic zoonosis. A cross‐sectional study was carried out to determine the exposure to T. gondii in equids in Europe. Serum samples from 1399 equids (1085 horses, 238 donkeys, and 76 mules/hinnies) bred in four European countries (Italy, Spain, United Kingdom [UK], and Ireland) were collected during the period of 2013–2021. The overall seroprevalence of T. gondii was 18.9% (95% Confidence Interval [CI]: 16.9–21.0) by using the modified agglutination test (MAT) at a cut‐off of 1:25. Seropositivity by country was 27.1% in Italy, 16.6% in Spain, 12.0% in UK and 7.0% in Ireland. Anti‐T. gondii antibodies were detected in 12.8% of the horses, 43.7% of the donkeys, and in 28.9% of the mules/hinnies. A Generalized Estimating Equation (GEE) analysis was carried out to study the associations between seropositivity and explanatory variables related to individuals, herds, and management measures on these herds, selected based on the bivariate analysis. The risk for being seropositive for T. gondii was 5.3 and 2.7 times higher in donkeys and mules/hinnies than in horses, respectively. In addition, significantly higher seropositivity was observed in horses from herds that used disinfectants less than once a week (13.9%; p = 0.038, odds ratio [OR] = 1.6; 95% CI: 1.03–2.62) compared with those from herds that performed weekly disinfection of the facilities (9.4%). This is the first large‐scale seroepidemiological study on T. gondii comprising horses, donkeys, and mules/hinnies in Europe and the first report of T. gondii exposure in horses from Ireland and UK. We found a widespread distribution of T. gondii among equid populations in different European countries. The seroprevalence found in these species, especially in donkeys and mules/hinnies, highlights the potential risk of human infection through the consumption of their raw/undercooked milk or meat. [ABSTRACT FROM AUTHOR]

Considering the objectives of "One Health" and the Sustainable development Goals "Good health and well-being" for the development of effective strategies to apply against bacterial resistance, food safety dangers, and zoonosis risks, this project explored the isolation and identification of Lactobacillus strains from the intestinal tract of recently weaned mice; as well as the assessment of antibacterial activity against clinical and zoonotic pathogens. For molecular identification, 16S rRNA gene-specific primers were used and, via BLAST-NCBI, 16 Ligilactobacillus murinus, one Ligilactobacillus animalis, and one Streptococcus salivarius strains were identified and registered in GenBank after the confirmation of their identity percentage and the phylogenetic analysis of the 16 Ligilactobacillus murinus strains and their association with Ligilactobacillus animalis. The 18 isolated strains showed antibacterial activity during agar diffusion tests against Listeria monocytogenes ATCC 15313, enteropathogenic Escherichia coli O103, and Campylobacter jejuni ATCC 49943. Electrophoretic and zymographic techniques confirmed the presence of bacteriolytic bands with a relative molecular mass of 107 kDa and another of 24 kDa in Ligilactobacillus murinus strains. UPLC-MS analysis allowed the identification of a 107 kDa lytic protein as an N-acetylmuramoyl-L-amidase involved in cytolysis and considered a bacteriolytic enzyme with antimicrobial activity. The 24 kDa band displayed similarity with a portion of protein with aminopeptidase function. It is expected that these findings will impact the search for new strains and their metabolites with antibacterial activity as an alternative strategy to inhibit pathogens associated with major health risks that help your solution. [ABSTRACT FROM AUTHOR]

Aim of this study was to comparatively evaluate antibacterial activities of methanol (MetOH), acetone (Ace), petroleum ether (PE) and aqueous (dw) leaf (L), root (R), and seed (S) extracts of Corchorus olitorius L. on both food- and plant-borne pathogens, with DPPH radical scavenging activities (DRSA), and quantitative and qualitative constituent analysis. Leaf PE has the highest strain susceptibility on both food- and plant-borne pathogens. Clavibacter michiganensis, Pseudomonas tomato, and Erwinia caratovora were susceptible to nearly all the leaf and seed extracts. Very low minimum inhibitory concentration (8-128 μg mL−1) and minimum bactericidal concentration (32-2048 μg mL−1) were determined for both leaf and seed extracts against C. michiganensis. Total phenolic contents were correlated to DRSA. The phenolic compounds tested were higher in the leaf MetOH, cholorogenic acid being the most abundant one. Palmitic acid was determined in leaf PE and seed PE extracts. Results presented here demonstrate high antibacterial activity of C. olitorius leaf seed extracts against phytopathogens for the first time, and provide the most comprehensive data on the antibacterial activity screening against food-borne pathogens. Considering limitations in plant disease control, antibacterial activities of these extracts would be important in plant disease control.

According to World Health Organization report, each year worldwide, unsafe food causes 600 million food-borne diseases and 420,000 deaths. Additionally, 30% of deaths occur in children under 5 years old. On the other hand, with the increase in people's awareness about nutritional value of food, the dietary trends have led to a continuously increasing demand for ready-to-eat foods. Hence, enhancing food safety is very important for the consumers. The application of modern food preservation methods is quite necessary in order to improve safety and extend shelf life with the lowest adverse effects on the sensory characteristics and nutritional value of seafood products. Non-thermal methods such as high pressure, ultrasonic, cold plasma techniques, and electrolyzed water have been developed to maintain food hygiene or disinfection. In recent years, the use of electrolyzed water (EW), which is obtained by electrolysis of dilute salt solution (NaCl) or hydrogen chloride (HCl), is known as an effective disinfection technique against a wide range of food pathogens. In this review, the effectiveness of using electrolyzed water in aquatic products, as well as their antimicrobial properties against food pathogens, have been investigated. [ABSTRACT FROM AUTHOR]

This study examined the chemical composition and antimicrobial activities of essential oils isolated from the leaves, twigs, and fruits of Neolitsea sericea var. aurata from Taiwan. The major compounds responsible for antimicrobial activity were isolated and identified. The leaf, twig, and fruit essential oils were obtained through hydrodistillation in a Clevenger-type apparatus and were characterized through gas chromatography (GC) with flame ionization detection (GC/FID) and GC/mass spectrometry (GC/MS). The major compounds were (E)-β-ocimene (49.3%) and sericenine (31.6%) in the leaf oil; (E)-β-ocimene (73.7%), α-cadinol (6.8%), and α-muurolol (4.0%) in the twig oil; and (E)-β-ocimene (84.7%) in the fruit oil. The twig oil exhibited the strongest antibacterial and antifungal activities. We isolated α-cadinol and α-muurolol, the main components of NTO4 fraction, and used them in antimicrobial tests; the two compounds exhibited excellent antimicrobial activities. The twig essential oil contains α-cadinol and α-muurolol and exhibited excellent antimicrobial activities against food-borne pathogens; thus, they are worthy of further research and development. [ABSTRACT FROM AUTHOR]

The problem of microbial biofilms has come to the fore alongside food, pharmaceutical, and healthcare industrialization. The development of new antibiofilm products has become urgent, but it includes bioprospecting and is time and money-consuming. Contemporary efforts are directed at the pursuit of effective compounds of natural origin, also known as "green" agents. Mushrooms appear to be a possible new source of antibiofilm compounds, as has been demonstrated recently. The existing modeling methods are directed toward predicting bacterial biofilm formation, not in the presence of antibiofilm materials. Moreover, the modeling is almost exclusively targeted at biofilms in healthcare, while modeling related to the food industry remains under-researched. The present study applied an Artificial Neural Network (ANN) model to analyze the anti-adhesion and anti-biofilm-forming effects of 40 extracts from 20 mushroom species against two very important food-borne bacterial species for food and food-related industries—Listeria monocytogenes and Salmonella enteritidis. The models developed in this study exhibited high prediction quality, as indicated by high r2 values during the training cycle. The best fit between the modeled and measured values was observed for the inhibition of adhesion. This study provides a valuable contribution to the field, supporting industrial settings during the initial stage of biofilm formation, when these communities are the most vulnerable, and promoting innovative and improved safety management. [ABSTRACT FROM AUTHOR]

Introduction: Due to the increasing resistance of bacteria and fungi to antimicrobials, it is necessary to search for effective alternatives to prevent and treat pathogens causing diseases in humans, animals, and plants. In this context, the mycosynthesized silver nanoparticles (AgNPs) are considered as a potential tool to combat such pathogenic microorganisms. Methods: AgNPs were synthesized from Fusarium culmorum strain JTW1 and characterized by Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), Fourier Transform Infrared (FTIR) spectroscopy, Nanoparticle Tracking Analysis (NTA), Dynamic Light Scattering (DLS) and Zeta potential measurement. The minimum inhibitory (MIC) and biocidal concentrations (MBC) were determined against 13 bacterial strains. Moreover, the combined effect of AgNPs with antibiotics (streptomycin, kanamycin, ampicillin, tetracycline) was also studied by determining the Fractional Inhibitory Concentration (FIC) index. The anti-biofilm activity was examined by crystal violet and fluorescein diacetate (FDA) assays. Furthermore, antifungal activity of AgNPs was evaluated against a panel of phytopathogenic fungi viz., Botrytis, Colletotrichum, Fusarium, Phoma, Sclerotinia, and an oomycete pathogen Phytophthora by agar well-diffusion and micro-broth dilution method to evaluate the minimal AgNPs concentrations that inhibit fungal spore germination. Results: Fungi-mediated synthesis resulted in the formation of small (15.56± 9.22 nm), spherical and stable (zeta potential of - 38.43 mV) AgNPs with good crystallinity. The results of FTIR spectroscopy indicated the presence of various functional groups, namely hydroxyl, amino, and carboxyl ones, from the biomolecules on the surface of AgNPs. The AgNPs showed antimicrobial and antibiofilm formation activities against Gram-positive and Gram-negative bacteria. The values of MIC and MBC ranged between 16-64 and 32-512 ㎍ mL-1, respectively. The enhanced effect of AgNPs in combination with antibiotics was confirmed against human pathogens. The highest synergistic effect (FIC = 0.0625) was demonstrated by the combination of AgNPs with streptomycin against two strains of Escherichia coli (ATCC 25922 and ATCC 8739), followed by Klebsiella pneumoniae and Pseudomonas aeruginosa (FIC = 0.125). Enhanced effects of AgNPs with ampicillin were also shown against Staphylococcus aureus ATCC 25923 (FIC = 0.125) and P. aeruginosa (FIC = 0.25), as well as kanamycin against S. aureus ATCC 6538 (FIC = 0.25). The crystal violet assay revealed that the lowest concentration of AgNPs (0.125 ㎍ mL-1) reduced the development of biofilms of Listeria monocytogenes and Salmonella enterica, while the maximum resistance was shown by Salmonella infantis, its biofilm was reduced after exposure to a concentration of 512㎍ mL-1. A high inhibitory effect on the activity of bacterial hydrolases was observed by the FDA assay. AgNPs at a concentration of 0.125 ㎍ mL-1 reduced the hydrolytic activity of all biofilms formed by the tested pathogens, except E. coli ATCC 25922, P. aeruginosa, and Pectobacterium carotovorum (efficient concentration was 2-fold higher, at 0.25 ㎍ mL-1), while the hydrolytic activity of E. coli ATCC 8739, Salmonella infantis and S. aureus ATCC 6538 was suppressed after treatment with AgNPs at concentrations of 0.5, 2 and 8㎍ mL-1, respectively. Moreover, AgNPs inhibited fungal growth and spore germination of Botrytis cinerea, Phoma lingam, and Sclerotinia sclerotiorum. MIC and MFC values of AgNPs against spores of these fungal strains were determined at 64, 256, and 32 ㎍ mL-1, and zones of growth inhibition were 4.93, 9.54, and 3.41 mm, respectively. Discussion: Fusarium culmorum strain JTW1 was found to be an eco-friendly biological system for an easy, efficient and inexpensive synthesis of AgNPs. In our study, the mycosynthesised AgNPs demonstrated remarkable antimicrobial (antibacterial and antifungal) and antibiofilm activities against a wide range of human and plant pathogenic bacteria and fungi singly and in combination with antibiotics. These AgNPs could be applied in medicine, agriculture, and food industry to control such pathogens that cause numerous human diseases and crop losses. However, before using them extensive animal studies are required to evaluate the toxicity, if any. [ABSTRACT FROM AUTHOR]

The present study aimed to characterize the mode of action of a novel antimicrobial peptide isolated from egg yolk hydrolysate. The EYHp6, KGGDLGLFEPTL, exhibited inhibition against Salmonella enterica serovar Typhimurium TISTR 292 and S. enterica serovar Enteritidis DMST 15679 with a MIC value of 2 mM. In contrast, S. enterica serovar Newport ATCC 6962 and other strains of Typhimurium and Enteritidis were inhibited at 4 mM. EYHp6 increased the cell membrane permeability of S. Typhimurium TISTR 292, leading to DNA leakage. Membrane integrity determined by propidium iodide and SYTO9 staining visualized by confocal microscopy demonstrated that EYHp6 at 1 × MIC induced disruption of cell membranes. Electron microscopy revealed that treatment of S. Typhimurium with EYHp6 led to damage to the cell membrane, causing the leakage of intracellular contents. Synchrotron-based Fourier-transform infrared spectroscopy indicated that EYHp6 killed S. Typhimurium by targeting fatty acids and nucleic acids in the cell membrane. The peptide did not show hemolytic activity up to 4 mM. These findings suggest that EYHp6 could be a promising antibacterial agent for controlling the growth of S. enterica.

Campylobacter and pathogenic Escherichia coliillnesses have been attributed to the consumption of fresh produce. The leafy green, kale, is increasingly consumed raw. In comparison to other leafy greens, kale has a longer shelf-life. Due to the extended shelf-life of kale, it is warranted to examine the survival of pathogenic Campylobacter jejuni and E. coli O157:H7 inoculated on the surface of kale stored in a controlled environment at 4 ± 1.4°C, and average humidity of 95 ± 1.9% over a 23-day period. At predetermined time points (days 0, 1, 2, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21), inoculated kale was destructively sampled and the surviving bacteria determined by serial dilution and plating onto Tryptic soy agar, Charcoal cefoperozone deoxycholate agar, and Eosin methylene blue for total aerobic bacteria, C. jejuni, and E. coli O157:H7, respectively. Enrichment and PCR were used for detection when pathogens were not detected using serial dilution and plating. Aerobic heterotrophic bacteria increased over the 23-day period, in contrast, significant declines in the inoculated pathogens were observed. Inoculated E. coli O157:H7 survived longer on kale (up to 19 d); in comparison, C. jejuni was undetectable by day 13 using enrichment and PCR or plating. In conclusion, C. jejuni and E. coli O157:H7 declined on fresh kale over time when held at refrigerated temperatures but were still detected during the majority of the time when the kale would likely still be considered edible by consumers.

IntroductionDue to the increasing resistance of bacteria and fungi to antimicrobials, it is necessary to search for effective alternatives to prevent and treat pathogens causing diseases in humans, animals, and plants. In this context, the mycosynthesized silver nanoparticles (AgNPs) are considered as a potential tool to combat such pathogenic microorganisms.MethodsAgNPs were synthesized from Fusarium culmorum strain JTW1 and characterized by Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), Fourier Transform Infrared (FTIR) spectroscopy, Nanoparticle Tracking Analysis (NTA), Dynamic Light Scattering (DLS) and Zeta potential measurement. The minimum inhibitory (MIC) and biocidal concentrations (MBC) were determined against 13 bacterial strains. Moreover, the combined effect of AgNPs with antibiotics (streptomycin, kanamycin, ampicillin, tetracycline) was also studied by determining the Fractional Inhibitory Concentration (FIC) index. The anti-biofilm activity was examined by crystal violet and fluorescein diacetate (FDA) assays. Furthermore, antifungal activity of AgNPs was evaluated against a panel of phytopathogenic fungi viz., Botrytis, Colletotrichum, Fusarium, Phoma, Sclerotinia, and an oomycete pathogen Phytophthora by agar well-diffusion and micro-broth dilution method to evaluate the minimal AgNPs concentrations that inhibit fungal spore germination.ResultsFungi-mediated synthesis resulted in the formation of small (15.56 ± 9.22 nm), spherical and stable (zeta potential of – 38.43 mV) AgNPs with good crystallinity. The results of FTIR spectroscopy indicated the presence of various functional groups, namely hydroxyl, amino, and carboxyl ones, from the biomolecules on the surface of AgNPs. The AgNPs showed antimicrobial and antibiofilm formation activities against Gram-positive and Gram-negative bacteria. The values of MIC and MBC ranged between 16–64 and 32–512 μg mL−1, respectively. The enhanced effect of AgNPs in combination with antibiotics was confirmed against human pathogens. The highest synergistic effect (FIC = 0.0625) was demonstrated by the combination of AgNPs with streptomycin against two strains of Escherichia coli (ATCC 25922 and ATCC 8739), followed by Klebsiella pneumoniae and Pseudomonas aeruginosa (FIC = 0.125). Enhanced effects of AgNPs with ampicillin were also shown against Staphylococcus aureus ATCC 25923 (FIC = 0.125) and P. aeruginosa (FIC = 0.25), as well as kanamycin against S. aureus ATCC 6538 (FIC = 0.25). The crystal violet assay revealed that the lowest concentration of AgNPs (0.125 μg mL−1) reduced the development of biofilms of Listeria monocytogenes and Salmonella enterica, while the maximum resistance was shown by Salmonella infantis, its biofilm was reduced after exposure to a concentration of 512 μg mL−1. A high inhibitory effect on the activity of bacterial hydrolases was observed by the FDA assay. AgNPs at a concentration of 0.125 μg mL−1 reduced the hydrolytic activity of all biofilms formed by the tested pathogens, except E. coli ATCC 25922, P. aeruginosa, and Pectobacterium carotovorum (efficient concentration was 2-fold higher, at 0.25 μg mL−1), while the hydrolytic activity of E. coli ATCC 8739, Salmonella infantis and S. aureus ATCC 6538 was suppressed after treatment with AgNPs at concentrations of 0.5, 2 and 8 μg mL−1, respectively. Moreover, AgNPs inhibited fungal growth and spore germination of Botrytis cinerea, Phoma lingam, and Sclerotinia sclerotiorum. MIC and MFC values of AgNPs against spores of these fungal strains were determined at 64, 256, and 32 μg mL−1, and zones of growth inhibition were 4.93, 9.54, and 3.41 mm, respectively.DiscussionFusarium culmorum strain JTW1 was found to be an eco-friendly biological system for an easy, efficient and inexpensive synthesis of AgNPs. In our study, the mycosynthesised AgNPs demonstrated remarkable antimicrobial (antibacterial and antifungal) and antibiofilm activities against a wide range of human and plant pathogenic bacteria and fungi singly and in combination with antibiotics. These AgNPs could be applied in medicine, agriculture, and food industry to control such pathogens that cause numerous human diseases and crop losses. However, before using them extensive animal studies are required to evaluate the toxicity, if any.

In this paper, we report a sensitive surface‐enhanced Raman scattering (SERS) biosensor for the detection of food‐borne pathogens on the basis of recognition of bacterial cells through the specific targeting agent vancomycin. Vancomycin‐functionalized gold nanostars (GNSs–4–MBA–van) were synthesized as SERS tags for specific and sensitive quantification of food pathogens. The Raman reporter molecule 4–mercaptobenzoic acid (4–MBA) was also a linker molecule to attach vancomycin to gold nanostars. The proposed vancomycin‐based SERS technique achieved a detection limit of 5.7 and 8.2 CFU/ml for Staphylococcus aureus and Escherichia coli, respectively. The logarithm of the bacterial concentration ranged from 101 to 107 CFU/ml and demonstrated a linear relationship with the SERS intensity of the characteristic 4–MBA Raman peak at 1,081 cm−1. Principle component analysis was used to discriminate the two pathogens simultaneously by using reproducible and specific intrinsic fingerprints and intracellular components obtained through SERS. Moreover, the spiking tests revealed that GNSs–4–MBA–van is effective as a potential biosensor for detecting pathogens in the fresh meat and food‐processing industries. [ABSTRACT FROM AUTHOR]

Background: The disinfection process represents an important activity closely linked to the removal of micro-organisms in common processing systems. Traditional disinfectants are often not sufficient to avoid the spread of food pathogens; therefore, innovative strategies for decontamination are crucial to countering microbial transmission. This study aims to assess the antimicrobial efficiency of tetrapotassium iminodisuccinic acid salt (IDSK) against the most common pathogens present on surfaces, especially in food-borne environments. Methods: IDSK was synthesized from maleic anhydride and characterized through nuclear magnetic resonance (NMR) spectroscopy (both 1H-NMR and 13C-NMR), thermogravimetric analysis (TGA) and Fourier Transform Infrared (FTIR) spectroscopy. The antibacterial activity was performed via the broth microdilution method and time-killing assays against Escherichia coli, Staphylococcus aureus, Salmonella enterica, Enterococcus faecalis and Pseudomonas aeruginosa (IDSK concentration range: 0.5–0.002 M). The biofilm biomass eradicating activity was assessed via a crystal violet (CV) assay. Results: The minimum inhibitory concentration (MIC) of IDSK was 0.25 M for all tested strains, exerting bacteriostatic action. IDSK also reduced biofilm biomass in a dose-dependent manner, reaching rates of about 50% eradication at a dose of 0.25 M. The advantages of using this innovative compound are not limited to disinfecting efficiency but also include its high biodegradability and its sustainable synthesis. Conclusions: IDSK could represent an innovative and advantageous disinfectant for food processing and workers' activities, leading to a better quality of food and safer working conditions for the operators. [ABSTRACT FROM AUTHOR]

Food safety is a significant public health issue in both developed and developing countries. Previous detection methods struggle to meet the current demands. We have proposed a new way to detect pathogens, allowing detection to be visualized by the naked eye. Using our newly developed assay, when target genes are present in the reaction, corresponding padlock probes form closed-loop molecules. Each reaction tube contains a pair of universal primers for identifying target genes. The ring padlock probes and corresponding universal primers start hyperbranched rolling circle amplification (HRCA) under the action of the polymerase, so as to gain branched chain amplification products, which are irreversibly entangled with magnetic particles to form aggregated magnetic particle clusters, and the detection results are visible to naked eyes. On the contrary, by using linear probes, the clustering of magnetic particles will not be produced. This method was applied to the detection of five food-borne pathogens enterohemorrhagic Escherichia coli (EHEC), enterotoxigenic Escherichia coli (ETEC), enteropathogenic Escherichia coli (EPEC), enteroinvasive Escherichia coli (EIEC) and Escherichia coli (E. coli), with detection limits of 1 × 103, 1 × 104, 1 × 103, 1 × 104 and 1 × 102 CFU/mL, respectively. This method can realize multiplex automatic detection of nucleic acid and shows great development potential in the field of molecular diagnosis. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is one of the most challenging public health problems worldwide, and integrated surveillance is a key aspect in a One Health control strategy. Additionally, Salmonella is the second most common zoonosis in Europe. We aimed to investigate the circulation of Salmonella strains and their related antimicrobial resistance in human, environmental, and wild boar samples from the northwest of Italy, from 2018 to 2020, to obtain a more comprehensive epidemiological picture. Salmonella Typhimurium 1,4,[5],12:i:-, S. Veneziana and S. Newport were the most common serotypes occurring in humans, the environment, and wild boar, respectively. Antimicrobial resistance was rather common in Salmonella isolates, with those from human displaying the highest degree of resistance against sulfadiazine–sulfamerazine–sulfamethazine (>90% of resistance). Moreover, resistance against azithromycin were exclusively observed in environmental samples, while only 7.7% (95% CI = 1.6–20.8) of wild boar isolates experienced resistance against trimethoprim–sulfamethoxazole. Multidrug resistance concurrently involved up to seven antimicrobial classes in human isolates, including third-generation cephalosporins and fluoroquinolones. Salmonella Typhimurium in humans and serotypes Goldcoast and Rissen from environmental sources showed the highest levels of resistance. This study shows diverse antimicrobial resistance patterns in Salmonella strains isolated from different sources and gives a broad picture of antimicrobial resistance spread in wild animals, humans, and the environment. [ABSTRACT FROM AUTHOR]

Background: The present study aimed to evaluate antimicrobial properties of corn starch edible films prepared with Bunium persicum essential-oil nanoemulsions (BPEOne) and BPEOne fortified with cinnamaldéhyde (Fortified BPEOne with CIN) against some common food-borne pathogens. Methods: Treatments (films containing BPEO, CIN, BPEOne, BPEO+CIN, BPEOne+CIN, Fortified BPEOne with CIN in 0.5, 1, 2, and 4 % concentrations) were prepared using starch solutions. In vitro antimicrobial activity of BPEOne films was evaluated using agar disk diffusion and plate count assay methods against Salmonella enteritidis, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus. Results: The inhibitory effect of each treatment against some food-borne pathogen bacteria increased significantly with the enhancement of concentrations (P<0.05). The highest bacterial inhibitory effect was achieved in higher concentrations (2%, 4%). The antimicrobial activity of BPEO nanoemulsion was higher than pure BPEO-containing films. S. aureus and S. enteritidis were more susceptible and resistant bacteria to BPEO, BPEO nanoemulsion-containing films, respectively. Conclusion: The films containing a combination of the BPEO (or its nanoemulsion) with CIN have better antimicrobial activity, however the fortification of nanoemulsions with other antimicrobial agents may not show the same results. [ABSTRACT FROM AUTHOR]

Alpha-phellandrene is a very common cyclic monoterpene found in several EOs, which shows extensive biological activities. Therefore, the main focus of the present systematic review was to provide a comprehensive and critical analysis of the state of the art regarding its biological activities and pharmaceutical and food applications. In addition, the study identified essential oils rich in alpha-phellandrene and summarized their main biological activities as a preliminary screening to encourage subsequent studies on their single components. With this review, we selected and critically analyzed 99 papers, using the following bibliographic databases: PubMed, SciELO, Wiley and WOS, on 8 July 2022. Data were independently extracted by four authors of this work, selecting those studies which reported the keyword "alpha-phellandrene" in the title and/or the abstract, and avoiding those in which there was not a clear correlation between the molecule and its biological activities and/or a specific concentration from its source. Duplication data were removed in the final article. Many essential oils have significant amounts of alpha-phellandrene, and the species Anethum graveolens and Foeniculum vulgare are frequently cited. Some studies on the above-mentioned species show high alpha-phellandrene amounts up to 82.1%. There were 12 studies on alpha-phellandrene as a pure molecule showed promising biological functions, including antitumoral, antinociceptive, larvicidal and insecticidal activities. There were 87 research works on EOs rich in alpha-phellandrene, which were summarized with a focus on additional data concerning potential biological activities. We believe this data is a useful starting point to start new research on the pure molecule, and, in particular, to distinguish between the synergistic effects of the different components of the OEs and those due to alpha-phellandrene itself. Toxicological data are still lacking, requiring further investigation on the threshold values to distinguish the boundary between beneficial and toxic effects, i.e., mutagenic, carcinogenic and allergenic. All these findings offer inspiration for potential applications of alpha-phellandrene as a new biopesticide, antimicrobial and antitumoral agent. In particular, we believe our work is of interest as a starting point for further studies on the food application of alpha-phellandrene. [ABSTRACT FROM AUTHOR]

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The purpose of this experiment was to study the antibacterial effect of saponins from different sources on common food-borne pathogens, and to develop the application of saponins in the field of food. The size of inhibition zone, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of saponins from three different sources (Gleditsia sinensis shell, green tea and Camellia oleifera seed) were determined by Oxford cup method and double dilution method, respectively. The results showed that the three saponins from different sources all had antibacterial effect on four common food-borne pathogenic bacteria, namely Bacillus subtilis, Staphylococcus aureus, Salmonella sp. and Escherichia coli. Among them, the saponins derived from Gleditsia sinensis shell had the best antibacterial effect. The double dilution method showed that their MICs for the two Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus were both 4 mg/mL, and the MBCs were 16 mg/mL and 8 mg/mL respectively; The MIC of Salmonella and E.coli of two Gram-negative bacteria were 4 mg/mL and 8 mg/mL, and MBC were 8 mg/mL and 16 mg/mL respectively. The results provide devidence for the research and development of safe, high effective and natural plant food preservatives.

Toxoplasma gondii is a worldwide occurring apicomplexan parasite. Due to its high seroprevalence in livestock as well as in game animals, T. gondii is an important food-borne pathogen and can have significant health implications for humans as well as for pets. This article describes the prevalence of T. gondii in free-ranging macropods hunted for consumption. All hunted macropod species (commercial as well as non-commercial hunt) show a positive seroprevalence for T. gondii. This seroprevalence is influenced by various factors, such as sex or habitat. Furthermore, the parasite shows a high level of genetic variability in macropods. Genetically variable strains have already caused outbreaks of toxoplasmosis in the past (Canada and the US). These were attributed to undercooked game meat like venison. Despite this risk, neither Australia nor New Zealand currently have food safety checks against foodborne pathogens. These conditions scan pose a significant health risk to the population. Especially, since cases of toxoplasmosis have already been successfully traced back to insufficiently cooked kangaroo meat in the past.

The healthy olive pickles are those ones which are made by using lactic acid bacteria (LAB) as starter and protective cultures. Three lactic acid bacteria (LAB) strains namely: Lactobacillus plantarum LPS10 (L. plantarum), Lactobacillus fermentum PP17 (L. fermentum) and Pediococcus acidilactici MH512904 (P. acidilactici) were used for inhibition of some food-borne bacterial pathogens such as Listeria monocytogenes LMG10470 (L. monocytogenes), Staphylococcus aureus ATCC25923 (S. aureus), Bacillus cereus ATCC14579 (B. cereus) and Escherichia coli ATCC25922 (E. coli) in Brain Heart Infusion Broth (BHIB) and during pickling of green olive fruits. Cell free supernatants (CFS) from LAB showed distinctive inhibition of the indicator bacterial pathogens used and the inhibitory activity was more pronounced against Gram positive bacteria than that found against Gram negative E. coli strain used; the inhibitory activity of CFS was more pronounced than that obtained by neutralized cell free supernatants (NCFS). Cultures of LAB were used for inhibition of the food-borne pathogens during pickling of green olive fruits. The food-borne bacterial cells grew in olive pickles brine (control) and their growth (CFU/mL) decreased in treated pickles samples and distinctive differences in growth values (CFU/mL) were observed between control and treated samples. CFS from L. plantarum affected target cells of both L. monocytogenes and E. coli and caused cell deformations, cell shrinkage and cell lysis as showed by Transmission Electron Microscopic (TEM) examinations.

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The effectiveness of current antimicrobial methods for addressing for food-borne Gram-positive pathogens has dropped with the emergence of resistant strains. Consequently, new methods for addressing Gram-positive strains have to be developed continuously. This includes establishing novel targets for antimicrobial discovery efforts. Eukaryotic choline kinases have been highly developed as drug targets for the treatment of cancer, rheumatoid arthritis, malaria and many other conditions and diseases. Recently, choline kinase (ChoK) has been proposed as a drug target for Gram-positive species generally. The aim of this work was to discover novel, natural sources of inhibitors for bacterial ChoK from tea extracts. We report the first natural bacterial ChoK inhibitor with antimicrobial activity against Streptococcus pneumoniae: quercetin. [ABSTRACT FROM AUTHOR]