Identifying Putative Biomarkers of Foodborne Pathogens Using a Metabolomic Approach.

Metabolomics is the study of low molecular weight biochemical molecules (typically <1500 Da) in a defined biological organism or system. In case of food systems, the term "food metabolomics" is often used. Food metabolomics has been widely explored and applied in various fields including food analysis, food intake, food traceability, and food safety. Food safety applications focusing on the identification of pathogen-specific biomarkers have been promising. This chapter describes a nontargeted metabolite profiling workflow using gas chromatography coupled with mass spectrometry (GC-MS) for characterizing three globally important foodborne pathogens, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica, from selective enrichment liquid culture media. The workflow involves a detailed description of food spiking experiments followed by procedures for the extraction of polar metabolites from media, the analysis of the extracts using GC-MS, and finally chemometric data analysis using univariate and multivariate statistical tools to identify potential pathogen-specific biomarkers.

Propidium monoazide (PMA) qPCR assay compared to the plate count method for quantifying the growth of Salmonella enterica serotypes in vacuum-packaged turkey breast combined with a mathematical modeling approach.

This study compares the plate count (PC) and the Propidium Monoazide-quantitative Polymerase Chain Reaction (PMA-qPCR) methods to assess the growth of a cocktail of three serotypes of Salmonella enterica (Heidelberg, Typhimurium, and Enteritidis) in cooked, sliced, and vacuum-packaged turkey breast (STB) under isothermal storage temperatures (8 °C-20 °C), using predictive models. Standard curves were developed for PMA-qPCR, demonstrating high efficiency (101%) and sensitivity, with quantification limits ranging from 1 to 2 log10 CFU/g for all temperatures studied. Comparative analysis revealed a significant correlation (R2 = 0.99; 95% CI) between the PC and PMA-qPCR methods; however, the agreement analysis indicated a mean difference (Bias) of -0.11 log10 CFU/g (p < 0.05), suggesting underestimation by the PC method. This indicates the presence of stressed or viable but nonculturable (VBNC) cells, detectable by PMA-qPCR but not by PC. The Baranyi and Roberts model showed a good ability to describe the behavior of S. enterica cocktail in STB for PC and PMA-qPCR data under all isothermal conditions. The exponential secondary model more accurately represented the temperature dependence of the maximum specific growth rate compared to the Ratkowsky square root model, with R2 values ≥ 0.984 and RMSE values ≤ 0.011 for both methods. These results suggest that combining PMA-qPCR with predictive modeling allows for a more accurate prediction of S. enterica growth, compared to PC method. In the event of cold chain disruptions of meat products, the use of PMA-qPCR method allow the quantification of VBNC cells, that can still pose a health risk to consumers, especially in ready-to-eat products.

The applicability of the SERS technique in food contamination testing - The detailed spectroscopic, chemometric, genetic, and comparative analysis of food-borne Cronobacter spp. strains.

Microorganisms assigned as Cronobacter are Gram-negative, facultatively anaerobic, bacteria widely distributed in nature, home environments, and hospitals. They can also be detected in foods, milk powder, and powdered infant formula (PIF). Additionally, as an opportunistic pathogen, Cronobacter may cause serious infections, sometimes leading to the death of neonates and infants. Thus, it is essential to test food products for the presence of Cronobacter spp. The currently used standard described in ISO 22964:2017 is a laborious method that could be easily replaced by surface-enhanced Raman scattering (SERS). Here, we demonstrate that SERS allows the identification of food-borne bacteria belonging to Cronobacter spp. based on their SERS spectra. For this purpose, twenty-six Cronobacter strains from different food samples were analyzed. Additionally, it was shown that it is possible to differentiate them from other closely related pathogens such as Salmonella enterica subsp. enterica, Escherichia coli, or Enterobacter spp. The SERS results were supported by principal component analysis (PCA), as well as and sequencing of 16S rRNA, rpoB and fusA genes. Last but not least, it was demonstrated that the cells of Cronobacter sakazakii may be easily separated from PIF using an appropriate filter, microfluidic chip, and dielectrophoresis (DEP) technique.

Fresh produce-associated foodborne disease outbreaks in Australia, 2001 to 2017.

Abstract: Fresh produce is an important source of foodborne outbreaks in Australia. Using descriptive analysis, we examined confirmed and suspected foodborne outbreaks associated with fresh produce in Australia recorded in the OzFoodNet outbreak register from 2001 to 2017. The outbreak register contains reports of foodborne disease outbreaks collected by OzFoodNet epidemiologists and public health officials. A fresh produce outbreak was defined as the occurrence of two or more cases of the same illness in which the investigation had implicated a common food and this food contained fresh produce. A total of 92 fresh produce outbreaks were reported, encompassing 3,422 reported illnesses, 446 hospitalisations and four deaths. Of these outbreaks, 76.1% (70/92) were caused by a known pathogen, with the majority caused by either Salmonella enterica (n = 30) or Norovirus (n = 29). Most outbreaks (77.2%; 71/92) were associated with consumption of foods containing multiple ingredients, some of which were not fresh produce. The largest outbreaks associated with a single fresh produce item included bean sprouts contaminated with S. enterica serovar Saintpaul (419 illnesses and 76 hospitalisations) and semi-dried tomatoes contaminated with Hepatitis A (372 illnesses and 169 hospitalisations). Restaurants (45.7%; n = 42/92) and commercial catering (15.2%: n = 14/92) were common settings for fresh produce outbreaks. Outbreaks occurred in all states and territories of Australia and in all seasons, with an increased frequency in the warmer months (September-May). Although the number of fresh produce-associated outbreaks did not seem to be increasing in Australia, integrated surveillance is needed to rapidly identify sources of infection due to the propensity of these outbreaks to be large and widespread.

Role of migratory birds as a risk factor for the transmission of multidrug resistant Salmonella enterica and Escherichia coli to broiler poultry farms and its surrounding environment.

Multidrug resistance (MDR) considered as global crisis facing poultry industry. Migratory birds play very important role in the dissemination of antimicrobial resistant pathogen during their fly way specially to poultry farms. Therefore, 750 samples from migratory birds and 300 samples from broiler chicken farms and its environment were collected during the winter seasons of five years (2019 to 2023). The samples were subjected to the isolation of Salmonella enterica and Escherichia coli with the detection of antimicrobial resistance (phenotypic and genotypic) with insight to the genetic similarity between the isolates from migratory birds and broiler chickens' farms. Different members of Enterobacteriaceae were isolated; Salmonella enterica, Escherichia coli, Citrobacter, Enterobacter, Klebsiella, Proteus, Providencia, Serratia, Hafnia. 298 (28.4%) of S. enterica strains belonging to 27 serovars. S. Typhimurium, S. Kentucky, S. Enteritidis and S. Shangani were the common 4 serotypes between migratory birds and farms. Meanwhile, we found 489 (46.6%) isolates of E. coli belonging to 24 serogroups and O91, O128, O26, O125, O55, O103 and O159 were the common 7serogroups between migratory birds and farms samples. The majority of Salmonella (91.6%; 274 out of 298) and E. coli (92%; 450 out of 489) were MDR. The MDRI range of Salmonella and E. coli was 0.08- 1.The genetic similarity between the isolates of migratory birds and broiler chicken farms were detected by ERICPCR and hot map. This study suggests the continuous applications of surveillance programs for migratory birds and biosecurity measures in poultry farms.

Genomic epidemiology of ceftriaxone-resistant non-typhoidal Salmonella enterica strain in China.

Non-typhoidal Salmonella (NTS) is one of the top causes of diarrhea worldwide. Ceftriaxone is commonly recommended as the initial treatment option for Salmonella infections due to its antibacterial effectiveness. The objective of this study was to investigate the molecular epidemiological characteristics of NTS and to compare the phenotypic and genotypic profiles of antimicrobial resistance in multidrug-resistant Salmonella strains by sequencing 329 NTS strains collected from a county-level hospital between 2018 and 2021. Multi-locus sequence typing (MLST), antimicrobial resistance genes and plasmid types were identified by BacWGSTdb 2.0 webserver. Phylogenetic analysis of all NTS strains was carried out using Snippy and Gubbins software. The transferability of ceftriaxone resistant plasmids was confirmed through plasmid conjugation assays, and verified by S1-PFGE-Southern blot assays. The predominant serotypes among all NTS strains were Typhimurium (161/329), Enteritidis (49/329) and London (45/329). The most common sequence type observed was ST34 (86/329), followed by ST19 (72/329) and ST11 (47/329). The antimicrobial resistance of Salmonella to a wide range of antimicrobials showed an overall increase. Out of these 37 (11.24%) ceftriaxone-resistant strains, with the majority of them (33/37) being blaCTX-M. The predominant plasmid types identified were IncHI2 (14/21) and IncI1 (6/21), ranging in size from 70 kb to 360 kb. The conjugation efficiency was calculated with the high conjugation efficiency of 1.1 × 10- 5 to 9.3 × 10- 2. The strains varied widely, ranging from 3 to 45,024 single nucleotide polymorphisms (SNPs). There are close linkages observed among the predominant lineage, with an average of 78 SNPs between each pair of ST34 strains. The findings contribute to our understanding of the transmission and resistance mechanisms of multidrug-resistant Salmonella, thereby facilitating the development of effective control strategies.

Antimicrobial resistance and clonal relationships of Salmonella enterica Serovar Gallinarum biovar pullorum strains isolated in China based on whole genome sequencing.

BACKGROUND: Pullorum disease is a serious problem in many countries. Caused by Salmonella enterica serovar Gallinarum biovar Pullorum (S. Pullorum), it creates huge economic losses in the poultry industry. Although pullorum disease has been well-controlled in many developed countries, it is still a critical problem in developing countries. However, there is still a lack of information on S. Pullorum strains isolated from different regions and sources in China. The objective of this study was to supply the antimicrobial resistance patterns and clonal relationships of S. Pullorum from breeder chicken farms. METHODS: In this study, a total of 114 S. Pullorum strains recovered from 11 provinces and municipalities in China between 2020 and 2021 were selected. These 114 S. Pullorum strains were analyzed using whole genome sequencing (WGS). Antimicrobial resistance (AMR) was tested both by genotypic prediction using the WGS method and using disc diffusion to assess phenotypic AMR. RESULTS: These 114 sequenced S. Pullorum strains were divided into three sequence types (STs), the dominant STs was ST92 (104/114). Further core genome multi-locus sequence typing analysis indicated that 114 S. Pullorum strains may have a close relationship, which could be clonally transmitted among different provinces and municipalities. Our results showed a close relationship between the S. Pullorum strains found in different regions, indicating these strains may have been transmitted in China a long time ago. Nearly all S. Pullorum strains 94.74% (n = 108) were resistant to at least one antimicrobial class, and 35.96% of the examined Salmonella strains were considered multiple drug resistant. CONCLUSION: Overall, this study showed that S. Pullorum strains in China have a close genetic relationship in terms of antimicrobial resistance, suggesting widespread clonal transmission.

A global genome dataset for Salmonella Gallinarum recovered between 1920 and 2024.

Salmonella enterica serovar Gallinarum (S. Gallinarum) is an avian-specific pathogen responsible for fowl typhoid, a severe systemic disease with high mortality in chickens. This disease poses a substantial burden to the poultry industry, particularly in developing countries like China. However, comprehensive genome datasets on S. Gallinarum are lacking. Here, we present the most extensive S. Gallinarum genome dataset, comprising 574 well-collated samples. This dataset consists of 366 genomes sequenced in our laboratory and 208 publicly available genomes, collected from various continents over the past century. Using in silico prediction, we categorized S. Gallinarum into three distinct biovars. Regarding antimicrobial resistance, 238 strains (41.5%) carried antimicrobial resistance genes (ARGs) with a total of 635 records, while 232 strains (40.4%) exhibited multi-drug resistance. Mobile genomic elements (MGEs) serve as critical drivers for ARGs. Our dataset includes 5,636 MGEs records, with most MGEs belonging to prophages and plasmids. This dataset expands our understanding of the genomic characteristics of S. Gallinarum, providing valuable resources for future genomic studies to improve disease management.

First report on whole genome sequencing and comparative genomics of Salmonella enterica serovar Abortusequi isolated from Donkey in India.

Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi) is a leading cause of abortion in equines that hinders the rapid growth of equine industry. S. Abortusequi infection in equids has re-emerged over last ten years. In the present study, S. Abortusequi was isolated and characterized from donkeys during an abortion storm in the southern peninsular region of India. Further, whole genome sequencing and phylogenomic analysis revealed that the present isolate was clustered among S. Abortusequi clade. The core genome MLST (cgMLST) analysis based on hierarchical clustering and single nucleotide polymorphism (SNP) core-genome dendrogram of the present isolate against 10 S. Abortusequi isolates revealed that the present isolate established a distinct clade compared to all previously reported isolates. A comparison of cgMLST and SNP analyses revealed the same clustering concordance between isolates. In addition, comparative genomics and phylogenetic analysis was carried out with six S. Abortusequi serovars showed a higher number of core genes than accessory genes. Further, comparative analysis of phenotype and genotype antimicrobial resistance revealed a concordance of 32% and discordance of 68% respectively.

Salmonella enterica virulence databases and bioinformatic analysis tools development.

Salmonella enterica, a prominent foodborne pathogen, contributes significantly to global foodborne illnesses annually. This species exhibits significant genetic diversity, potentially impacting its infectivity, disease severity, and antimicrobial resistance. Whole genome sequencing (WGS) offers comprehensive genetic insights that can be utilized for virulence assessment. However, existing bioinformatic tools for studying Salmonella virulence have notable limitations. To address this gap, a Salmonella Virulence Database with a non-redundant, comprehensive list of putative virulence factors was constructed. Two bioinformatic analysis tools, Virulence Factor Profile Assessment and Virulence Factor Profile Comparison tools, were developed. The former provides data on similarity to the reference genes, e-value, and bite score, while the latter assesses the presence/absence of virulence genes in Salmonella isolates and facilitates comparison of virulence profiles across multiple sequences. To validate the database and associated bioinformatic tools, WGS data from 43,853 Salmonella isolates spanning 14 serovars was extracted from GenBank, and WGS data previously generated in our lab was used. Overall, the Salmonella Virulence database and our bioinformatic tools effectively facilitated virulence assessment, enhancing our understanding of virulence profiles among Salmonella isolates and serovars. The public availability of these resources will empower researchers to assess Salmonella virulence comprehensively, which could inform strategies for pathogen control and risk evaluations associated with human illnesses.

Assessing antimicrobial resistance profiles of Salmonella enterica in the pork production system.

Introduction. Salmonella enterica is a significant enteric pathogen affecting human and livestock health. Pork production is a common source of Salmonella contamination, with emerging multidrug resistance (MDR) posing a global health threat.Gap statement. Salmonella contamination and antimicrobial resistance (AMR) profiles in the pig production chain are underreported.Aim. To investigate the prevalence of S. enterica in the pig production chain and characterise their AMR profiles.Methodology. We collected 485 samples from pig farms, a standard pig abattoir and retail markets in Patthalung and Songkhla provinces in southern Thailand. Antimicrobial susceptibility testing was performed on these samples, and AMR profiles were determined.Results. S. enterica was detected in 68.67% of farm samples, 45.95% of abattoir samples and 50.67% of retail market samples. Analysis of 264 isolates, representing 18 serotypes, identified S. enterica serotype Rissen as the most prevalent. The predominant resistance phenotypes included ampicillin (AMP, 91.29%), tetracycline (TET, 88.26%) and streptomycin (STR, 84.47%). Over 80% of isolates showed resistance to three or more antimicrobial classes, indicating MDR. The AMP-STR-TET resistance pattern was found in nearly 70% of all MDR isolates across the production chain.Conclusions. The high prevalence of MDR is consistent with extensive antimicrobial use in the livestock sector. The presence of extensively resistant S. enterica highlights the urgent need for antimicrobial stewardship. Strengthening preventive strategies and control measures is crucial to mitigate the risk of MDR Salmonella spreading from farm to fork.

Prevalence, antimicrobial resistance, and genomic characterization of Salmonella strains isolated in Hangzhou, China: a two-year study.

This study explored the molecular epidemiology and resistance mechanisms of 271 non-duplicate Salmonella enterica (S. enterica) strains, isolated mainly from adults (209/271) in a tertiary hospital in Hangzhou between 2020 and 2021. Through whole-genome sequencing and bioinformatics, the bacterial strains were classified into 46 serotypes and 54 sequence types (ST), with S. Enteritidis, S. 1,4,[5],12:i:-, and S. Typhimurium being the most prevalent serotypes and ST11, ST34, and ST19 the most common STs. The strains isolated from adults were primarily S. Enteritidis (59/209), while from children were mainly S. 1,4,[5],12:i:- (20/62). Worryingly, 12.55% strains were multi-drug resistant (MDR), with resistance rates to cefepime (FEP), ceftazidime (CAZ), ceftriaxone (CRO) and cefotaxime (CTX) of 7.38%, 9.23%, 15.87% and 16.24%, respectively, and resistance rates to levofloxacin (LEV) and ciprofloxacin (CIP) of 8.49% and 19.19%, respectively. It is worth noting that the resistance rates of CRO and CTX in children reached 30.65%. A total of 34 strains carried extended-spectrum ß-lactamase (ESBL) genes, dominated by blaCTX-M-65 (13/34) and blaCTX-M-55 (12/34); it is notable that one strain of S. Saintpaul carried both blaCTX-M-27 and blaCTX-M-55. The resistance mechanism to cephalosporins was mainly due to ESBL genes (20/43), and other genes included AmpC and ß-lactamase genes. The strains resistant to quinolones mainly carried qnrS1 (27/53), and others included qnrB6, aac(6')-Ib-cr, and mutations in gyrA and parC. One strain did not carry common quinolone resistance genes but had a parC (p.T57S) mutation to cause CIP resistance. This research provides vital insights into the molecular epidemiology and resistance mechanisms of clinical S. enterica, implicating possible infection control strategies.

Integration of surface swab with optical microscopy for detection and quantification of bacterial cells from stainless-steel surfaces.

Swab sampling is a common method for recovering microbes on various environmental surfaces. Its successful application for a specific target depends on the proper swab method and the following detection assay. Herein, we evaluated critical factors influencing surface swab sampling, aiming to achieve the optimal detection and quantification performance of optical detection for bacterial cells on stainless-steel surfaces. Our results showed the recovery rate of Salmonella enterica (SE1045) cells from the 10 × 10 cm2 stainless-steel surface reached up to 92.71 ± 2.19% when using ammonia bicarbonate-moistened polyurethane foam swabs for gentle collection, followed by ultrasound-assisted release in NH4HCO3 solution. Among the six different foam swabs, the Puritan™ Sterile Large Foam Swab contributed the lowest background noise and highest recovery efficiency when integrated with the optical detection assay. Notably, our method exhibited a strong linear relationship (r2 = 0.9983) between the detected cell numbers and the theoretical number of SE1045 cells seeded on surfaces in the range of 104-107 Colony Forming Units (CFU), with a limit of detection of 7.2 × 104 CFU 100 cm-2. This integration was completed within 2 h, exhibiting the applicable potential in various settings.

Genome-wide fitness analysis of Salmonella enterica reveals aroA mutants are attenuated due to iron restriction in vitro.

Salmonella enterica is a globally disseminated pathogen that is the cause of over 100 million infections per year. The resulting diseases are dependent upon host susceptibility and the infecting serovar. As S. enterica serovar Typhimurium induces a typhoid-like disease in mice, this model has been used extensively to illuminate various aspects of Salmonella infection and host responses. Due to the severity of infection in this model, researchers often use strains of mice resistant to infection or attenuated Salmonella. Despite decades of research, many aspects of Salmonella infection and fundamental biology remain poorly understood. Here, we use a transposon insertion sequencing technique to interrogate the essential genomes of widely used isogenic wild-type and attenuated S. Typhimurium strains. We reveal differential essential pathways between strains in vitro and provide a direct link between iron starvation, DNA synthesis, and bacterial membrane integrity.IMPORTANCESalmonella enterica is an important clinical pathogen that causes a high number of deaths and is increasingly resistant to antibiotics. Importantly, S. enterica is used widely as a model to understand host responses to infection. Understanding how Salmonella survives in vivo is important for the design of new vaccines to combat this pathogen. Live attenuated vaccines have been used clinically for decades. A widely used mutation, aroA, is thought to attenuate Salmonella by restricting the ability of the bacterium to access particular amino acids. Here we show that this mutation limits the ability of Salmonella to acquire iron. These observations have implications for the interpretation of many previous studies and for the use of aroA in vaccine development.

Synthesis and pharmacological evaluation of Andrographolide and Ajwain as promising alternatives to antibiotics for treating Salmonella gallinarum infection in chicken.

The emergence of antibiotic resistance in pathogenic bacteria, including Salmonella gallinarum, poses a significant challenge to poultry health and food safety. In response, alternative strategies are urgently needed to mitigate bacterial infections without exacerbating antibiotic resistance. Phytoremediation, a sustainable and environmentally friendly approach, harnesses the natural detoxification capabilities of plants to remediate contaminants. This study explores the potential of combined phytoremediation using Andrographolide, derived from Andrographis paniculata, and Ajwain derived from Trachyspermum ammi as promising alternatives to antibiotics for treating Salmonella gallinarum infection in poultry. Andrographolide, known for its potent antimicrobial properties, exhibits inhibitory effects while Ajwain, rich in bioactive compounds, possesses antimicrobial and immunomodulatory properties. By leveraging their combined phytoremediation potential, Andrographolide and Ajwain offer a multifaceted approach to combat Salmonella gallinarum within the poultry environment. The study employed a rigorous experimental design, including in vitro assessments of antimicrobial susceptibility, cytotoxicity, and optimal concentration determination. Following this, in vivo experiments were conducted using a chicken model infected with Salmonella gallinarum. Results demonstrated that the selected combinations effectively reduced mortality rates, alleviated clinical symptoms, and mitigated gross pathological signs associated with Salmonella infection. Gene expression studies indicated a downregulation of proinflammatory cytokines, underscoring potential implications of a combined phytoremediation strategy as an innovative and sustainable solution to address Salmonella gallinarum infections in poultry production systems.

Optical detection and enumeration of Escherichia coli and Salmonella enterica using a low-magnification light microscope.

A rapid and cost-effective method for detecting bacterial cells from surfaces is critical to food safety, clinical hygiene, and pharmacy quality. Herein, we established an optical detection method based on a gold chip coating with 3-mercaptophenylboronic acid (3-MPBA) to capture bacterial cells, which allows for the detection and quantification of bacterial cells with a standard light microscope under low-magnification (10×) objective lens. Then, integrate the developed optical detection method with swab sampling to detect bacterial cells loading on stainless-steel surfaces. Using Salmonella enterica (SE1045) and Escherichia coli (E. coli OP50) as model bacterial cells, we achieved a capture efficiency of up to 76.0 ± 2.0 % for SE1045 cells and 81.1 ± 3.3 % for E. coli OP50 cells at 103 CFU/mL upon the optimized conditions, which slightly decreased with the increasing bacterial concentrations. Our assay showed good linear relationships between the concentrations of bacterial cells with the cell counting in images in the range of 103 -107 CFU/mL for SE1045, and 103 -108 CFU/mL for E. coli OP50 cells. The limit of detection (LOD) was 103 CFU/mL for both SE1045 and E. coli OP50 cells. A further increase in sensitivity in detecting E. coli OP50 cells was achieved through a heat treatment, enabling the LOD to be reduced as low as 102 CFU/mL. Furthermore, a preliminary application succeeded in assessing bacterial contamination on stainless-steel surfaces following integration with the approximately 40 % recovery rate, suggesting prospects for evaluating the bacteria from surfaces. The entire process was completed within around 2 h, costing merely a few dollars per sample. Considering the low cost of standard light microscopes, our method holds significant potential for practical industrial applications in bacterial contamination control on surfaces, especially in low-resource settings.

Biotransformation of Sumatriptan by Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Salmonella enterica subsp. enterica.

This study aimed at the biotransformation of sumatriptan by Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Salmonella enterica subsp. enterica and the identification of the drug metabolites by liquid chromatography-mass spectrometry. The drug was incubated with the organisms in tryptic soya broth at 37 °C. The broth was filtered and subjected to liquid chromatography-mass spectrometry. The metabolites identified by the use of mass spectral (+ve ion mode) fragmentation patterns were (3-methylphenyl)methanethiol (Bacillus subtilis), 1-(4-amino-3-ethylphenyl)-N-methylmethanesulfonamide (Salmonella enterica subsp. enterica) and 1-{4-amino-3-[(1E)-3-(dimethylamino)prop-1-en-1-yl]phenyl}methanesulfinamide (Salmonella enterica subsp. enterica, Bacillus subtilis, Pseudomonas aeruginosa, Staphylococcus aureus). These metabolites exhibit high gastrointestinal absorption, no blood-brain barrier permeability (except (3-methylphenyl)methanethiol), a bioavailability score of 0.55 and no inhibitory effect on CYP2C19, CYP2C9, CYP2D6, CYP3A4 or cytochrome P450 1A2 (except (3-methylphenyl)methanethiol), as determined by SwissADME software ver. 2024. The metabolites appear to be more toxic than the parent drug, as suggested by their calculated median lethal dose values. All four organisms under investigation transformed sumatriptan to different chemical substances that were more toxic than the parent drug.

High-contrast imaging of cellular non-repetitive drug-resistant genes via in situ dead Cas12a-labeled PCR.

In situ imaging of genes of pathogenic bacteria can profile cellular heterogeneity, such as the emergence of drug resistance. Fluorescence in situ hybridization (FISH) serves as a classic approach to image mRNAs inside cells, but it remains challenging to elucidate genomic DNAs and relies on multiple fluorescently labeled probes. Herein, we present a dead Cas12a (dCas12a)-labeled polymerase chain reaction (CasPCR) assay for high-contrast imaging of cellular drug-resistant genes. We employed a syncretic dCas12a-green fluorescent protein (dCas12a-GFP) to tag the amplicons, thereby enabling high-contrast imaging and avoiding multiple fluorescently labeled probes. The CasPCR assay can quantify quinolone-resistant Salmonella enterica in mixed populations and identify them isolated from poultry farms.

Prevalence of streptomycin and tetracycline resistance and increased transmissible third-generation cephalosporin resistance in Salmonella enterica isolates derived from food handlers in Japan from 2006 to 2021.

AIMS: The increasing prevalence of AmpC ß-lactamase (AmpC)- and extended-spectrum ß-lactamase (ESBL)- producing food pathogens is a serious public health concern. AmpC- and ESBL-producing Salmonella species pose a high risk of food contamination. This study aimed to investigate changes in the prevalence of Salmonella among food handlers in Japan from 2006 to 2021 using 100 randomly selected isolates from 2006, 2012, 2018, and 2021 with different serotypes and antimicrobial resistance patterns. METHODS AND RESULTS: The average Salmonella isolation rate was 0.070% (19 602/27 848 713). Serotyping revealed that the most common serotypes were Enteritidis in 2006, Infantis in 2012, Agoueve/Cubana in 2018, and Schwarzengrund in 2021. Antimicrobial susceptibility testing showed that Salmonella isolates exhibited the highest resistance to streptomycin (<40%), followed by tetracycline (<20%-40%). Moreover, 6% of the Salmonella isolates produced cephalosporinases with the blaCMY-2, blaCTX-M-14, and blaTEM genes. The annual incidence of cephalosporin resistance has increased. Plasmid conjugation assays revealed that cephalosporin-resistant Salmonella spp. transmitted their resistance to Escherichia coli. Additionally, plasmid genome analysis showed that the insertion sequence IS26 was encoded in the upstream and downstream regions of blaCTX-M-14 and qnrS1 in the IncHI1 plasmid, which could be transmitted to other bacteria. CONCLUSIONS: The tested Salmonella isolates showed high resistance to specific antibiotics, with differences in resistance depending on the serotype. Further increase and spread of transmissible cephalosporin-resistant strains should be noted.