Identification Of Foodborne Pathogens Research Articles

Rapid and visual detection of Listeria monocytogenes based on polymerase spiral reaction in fresh-cut fruit

Widespread foodborne pathogen Listeria monocytogenes exhibits a pronounced fatality rate among individuals with compromised immune systems, the elderly, and pregnant women. A simple and user-friendly approach is required to identify L. monocytogenes. A distinctive thermostatic nucleic acid amplification technology called polymerase spiral reaction (PSR) has been extensively employed in the identification of foodborne pathogens. In the study, a straightforward and intuitive approach for identifying L. monocytogenes in fresh-cut fruit was established using PSR in conjunction with hydroxy naphthol blue (HNB). Following optimization, the system's primary constitutes-betaine, dNTP, and MgSO4 were found to be 0.5 M, 1.0 mM, and 6 mM, respectively. Six strains of non- L. monocytogenes were used to test the assay's specificity. The PSR assay performed at 65 °C for 50 min demonstrated the capability to detect L. monocytogenes at level as low as 1 × 10−4 ng/μL DNA per tube and 5.1 × 101 CFU/g of freshly cut fruit. Notably, the sensitivity of this approach surpassed that of PCR by10-fold (1 × 10−3 ng/μL). In order to detect L. monocytogenes, the PSR assay was designed to be rapid, accurate, effective, and apparatus free. This innovative assay has provided researchers a fresh perspective on identification harmful microorganisms.

Pathogenic Escherichia coli (E. coli) food borne outbreak: Detection methods and controlling measures

Food-borne illnesses and diseases are major threats to human health. Common food borne pathogens include bacteria such as Escherichia coli Serogroup (O157), Bacillus cereus, Clostridium botulinum, Campylobacter sp., C. perfringens, some Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococcus aureus, and Vibrio spp. etc. Pathogenic E. coli infection usually causes severe diarrhea. Escherichia coli is the most common cause of acute urinary tract infections as well as urinary tract sepsis. Diarrheagenic E. coli pathotypes can be passed in the feces of humans and other animals. Safeguarding public health during mass gatherings is a big challenge. Despite the recent advances in food preservation techniques and food safety, significant disease outbreaks linked to food borne pathogens such as bacteria, fungi, and viruses still occur worldwide indicating that these pathogens still constitute significant risks to public health. However, at present the conventional method for E. coli detection requires several days, especially in cases where E. coli concentrations are low. Enrichment is a commonly used method for bacterial isolation to increase the cell counts of target bacteria. However, traditional culturing method is also a slow and laborious process requiring a series of steps and may require the use of adjunct methods (e.g. biochemical, serological, nucleic acid-based methods for conclusive identification) and can take up to a week for bacteria. The culture-based approaches coupled with other methods such as PCR, immunoassays, bacteriophages, NGS, Biosensors, and MALDI TOF MS are increasingly being used for the detection and identification of food borne pathogens. Nucleic acid-based methods such as Real-Time PCR and vPCR combined with sequencing approaches are more widely used than immunoassay and NGS-based approaches for pathogen detection. Nanobiotechnology is the latest approach for the detection of pathogens. Bacteriophages are one such unique biological entity that showed excellent host selectivity and have been actively used as recognition probes for pathogen detection. Antibiotics are efficient, powerful, and the most commonly used treatment against pathogenic E. coli. Other ways to treat or prevent E. coli contamination, such as probiotics, antimicrobial nanoparticles, and radiation treatment had been reported.

Foodborne Bacterial Pathogens in Animal, Food, and Environmental Samples Collected From the Physical Exposure of Children With Diarrhea in Ethiopia: A One Health Approach.

Foodborne pathogens (FBPs) are transmitted principally through the consumption of contaminated food or drinking water and pose a remarkable public health risk, particularly in low-income countries. A cross-sectional study was conducted between November 2021 and January 2023 to estimate the prevalence, co-occurrence, and monthly patterns of FBPs in the physical exposures of children with diarrhea in Harar town and Kersa district, Ethiopia. Animal, food, and environmental samples were collected from direct or indirect contact sites of children with diarrhea. The isolation and identification of FBPs, including nontyphoidal Salmonella (NTS), diarrheagenic E. coli (DEC), and Shigella, was performed using selective and deferential culture media and a series of biochemical tests. Among the 438 analyzed samples, the overall prevalence of these pathogens was 18.3%, with 3.9% co-occurrence and 14.4% single pathogen occurrence rates. The highest prevalence was observed in wastewater (40.9%; AOR = 3.3; 95% CI: 1.1-10.1). The pathogen detection rate in food was 17.9% (AOR = 1.2; 95% CI: 0.4-3.6), with no significant difference between animal-sourced and other food categories. The occurrence rates of NTS, DEC, and Shigella in the meat samples were 13.9%, 5.4%, and 6.5%, respectively. Interestingly, DEC and Shigella were detected in cooked food. Moreover, Shigella was detected in drinking water (5%) and other water sources (10%). A significantly higher prevalence of FBPs was detected in poultry than in cattle and camel feces. This study revealed fluctuations in the monthly occurrence patterns of FBPs, with a peak of 37.1% during the dry season. In conclusion, the study revealed a high prevalence of FBPs, with no significant differences between rural and urban areas or food and water sources, highlighting the need for food safety measures in both settings. Further studies with larger sample sizes and advanced diagnostics are recommended to determine the relative contribution of each source.

Novel next generation sequencing panel method for the multiple detection and identification of foodborne pathogens in agricultural wastewater.

Detecting and identifying the origins of foodborne pathogen outbreaks is a challenging. The Next-Generation Sequencing (NGS) panel method offers a potential solution by enabling efficient screening and identification of various bacteria in one reaction. In this study, new NGS panel primer sets that target 18 specific virulence factor genes from six target pathogens (Bacillus cereus, Yersinia enterocolitica, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus) were developed and optimized. The primer sets were validated for specificity and selectivity through singleplex PCR, confirming the expected amplicon size. Crosscheck and multiplex PCR showed no interference in the primer set or pathogenic DNA mixture. The NGS panel analysis of spiked water samples detected all 18 target genes in a single reaction, with pathogen concentrations ranging from 108 to 105 colony-forming units (CFUs) per target pathogen. Notably, the total sequence read counts from the virulence factor genes showed a positive association with the CFUs per target pathogen. However, the method exhibited relatively low sensitivity and occasional false positive results at low pathogen concentrations of 105 CFUs. To validate the detection and identification results, two sets of quantitative real-time PCR (qPCR) analyses were independently performed on the same spiked water samples, yielding almost the same efficiency and specificity compared to the NGS panel analysis. Comparative statistical analysis and Spearman correlation analysis further supported the similarity of the results by showing a negative association between the NGS panel sequence read counts and qPCR cycle threshold (Ct) values. To enhance NGS panel analysis for better detection, optimization of primer sets and real-time NGS sequencing technology are essential. Nonetheless, this study provides valuable insights into applying NGS panel analysis for multiple foodborne pathogen detection, emphasizing its potential in ensuring food safety.

Conventional and advanced detection techniques of foodborne pathogens: A comprehensive review

Foodborne pathogens are a major public health concern and have a significant economic impact globally. From harvesting to consumption stages, food is generally contaminated by viruses, parasites, and bacteria, which causes foodborne diseases such as hemorrhagic colitis, hemolytic uremic syndrome (HUS), typhoid, acute, gastroenteritis, diarrhea, and thrombotic thrombocytopenic purpura (TTP). Hence, early detection of foodborne pathogenic microbes is essential to ensure a safe food supply and to prevent foodborne diseases. The identification of foodborne pathogens is associated with conventional (e.g., culture-based, biochemical test-based, immunological-based, and nucleic acid-based methods) and advances (e.g., hybridization-based, array-based, spectroscopy-based, and biosensor-based process) techniques. For industrial food applications, detection methods could meet parameters such as accuracy level, efficiency, quickness, specificity, sensitivity, and non-labor intensive. This review provides an overview of conventional and advanced techniques used to detect foodborne pathogens over the years. Therefore, the scientific community, policymakers, and food and agriculture industries can choose an appropriate method for better results.

Advances in Traceability Typing and Identification of Foodborne Pathogens

Advances in Traceability Typing and Identification of Foodborne Pathogens

Monitoring of Microbial Safety of Foods Using Lectins: A Review

Food-borne diseases are on the rise, and these will likely continue as a public health concern into the coming decades. Majority of foodborne outbreaks are linked to infections by emerging foodborne pathogens such as Campylobacter, Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7. Foodborne pathogen identification becomes crucial in such scenarios to control these pathogens, associated outbreaks, and diseases. Pathogen detection systems have evolved as essential food safety tools to combat microbial threats and experts are striving to develop robust, accurate and ergonomic rapid pathogen-detection kits. Lectin, a ubiquitous biomolecule (sugar binding proteins) present in almost all domains of life is a promising alternative to molecular based methods as a bio-recognition molecule in detection of foodborne pathogens for biosensor applications, owing to its multivalency and spatial organization of ligands. Due to their extensive prevalence, lectin-based biosensors have become the most sought-after bio-recognition molecules in biosensor applications because of increased sensitivity and reduced cost when compared to immune-based biosensors. The current paper discusses the claimed benefits of lectin as a superior bio-recognition molecule, as well as its numerous applications in biosensor creation.

The food microbiological analyst: pairing tradition with the future

Choosing methods for the detection, isolation and identification of foodborne pathogens or spoilage microorganisms from foodstuffs is a complex task. Although there are standard food microbiology methods available around the world, many have changed little in decades, while some tests take many days to perform. The use of automated technologies, genomics and rapid methods are now replacing many traditional tests. Food microbiology analysts need to understand the underlying science and limitations of these methods for food analysis and the crucial importance of validation protocols for correct results reported to customers. This article discusses the current status of food microbiology analysis in commercial laboratories, what the future food microbiology analytic laboratory looks like, and the education changes that may be required to become a future food microbiology analyst.

Emerging Role of Biosensors for Detection of Foodborne Pathogens

Numerous techniques are employed for the detection of various types of pathogens in different foods. Rapid detection of foodborne pathogens is very important by using biosensors. The objectives of this review are to highlight the application of the biosensors in detecting foodborne pathogens, future trends of biosensors technology in foodborne pathogens detection, and limitations of biosensors in the food industry. Biosensors are analytical devices and one of the rapid detection methods that can combat certain limitations of conventional detection techniques, such as portability, time, cost, sensitivity, and others. Biosensors have many applications in different fields. Biosensors in the food industry are used for the detection and identification of foodborne pathogens. The uses of biosensors in food are increasing from day to day due to different factors. In the future, the latest generations, which are autonomous and nanomaterial-based biosensors will emerge. There is also a need to develop a biosensor that is easy to use and has a short detection time.

Identification of foodborne pathogens and antibacterial activity of ethanolic extract of Alangium salviifolium

Identification of foodborne pathogens and antibacterial activity of ethanolic extract of Alangium salviifolium

A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens.

Foodborne infections continue to plague Europe. Food safety monitoring is in crisis as the existing techniques for detecting pathogens do not keep up with the global rising of food production and consumption. Thus, the development of innovative techniques for detecting and identifying pathogenic bacteria has become critical. The aim of the present study was firstly to develop an innovative simple and low cost method of extracting bacterial DNA from contaminated food and water samples with Salmonella enteric(a) subsp. enteric(a) serovar Typhimurium and Listeria monocytogenes and its comparison with two commercial DNA extraction kits (Qiagen, Macherey-Nagel). Finally, pathogens' detection using two molecular techniques (PCR-electrophoresis, LAMP), in order to evaluate the best combination of DNA extraction and identification based on their sensitivity, cost, rapidity and simplicity. Considering the above criteria, among them, best was proved an in-house bacterial DNA extraction method, based on the chloroform-isoamyl alcohol protocol, with certain modifications. This technique showed statistically similar results in terms of sensitivity, compared to the commercial kits, while at the same time maintained high rapidity and much lower cost. Lastly, between the molecular techniques, LAMP was found more promising considering its simplicity, high rapidity and sensitivity. Conclusively, the in-house DNA extraction method along with the LAMP technique, was proven to be the best among the presented combinations.

Surface-Enhanced Raman Scattering (SERS) With Silver Nano Substrates Synthesized by Microwave for Rapid Detection of Foodborne Pathogens.

Rapid and sensitive methods have been developed to detect foodborne pathogens, a development that is important for food safety. The aim of this study is to explore Surface-enhanced Raman scattering (SERS) with silver nano substrates to detect and identify the following three foodborne pathogens: Escherichia coli O157: H7, Staphylococcus aureus and Salmonella. All the cells were resuspended with 10 mL silver colloidal nanoparticles, making a concentration of 107 CFU/mL, and were then exposed to 785 nm laser excitation. In this study, the results showed that all the bacteria can be sensitively and reproducibly detected directly by SERS. The distinctive differences can be observed in the SERS spectral data of the three food-borne pathogens, and the silver colloidal nanoparticles can be used as highly sensitive SERS-active substrates. In addition, the assay time required only a few minutes, which indicated that SERS coupled with the silver colloidal nanoparticles is a promising method for the detection and characterization of food-borne pathogens. At the same time, principle component analysis (PCA) and hierarchical cluster analysis (HCA) made the different bacterial strains clearly differentiated based on the barcode spectral data reduction. Therefore, the SERS methods hold great promise for the detection and identification of food-borne pathogens and even for applications in food safety.

Biotin-exposure-based immunomagnetic separation coupled with nucleic acid lateral flow biosensor for visibly detecting viable Listeria monocytogenes

Infectious diseases caused by Listeria monocytogenes pose a great threat to public health worldwide. Therefore, a rapid and efficient method for L. monocytogenes detection is needed. In this study, a biotin-exposure-based immunomagnetic separation (IMS) method was developed. That is, biotinylated antibody was first targeted to L. monocytogenes. Then, streptavidin-functionalized magnetic nanoparticles were added and anchored onto L. monocytogenes cells indirectly through the strong noncovalent interaction between streptavidin and biotin. Biotin-exposure-based IMS exhibited an excellent capability to enrich L. monocytogenes. Specifically, more than 90% of L. monocytogenes was captured when the bacterial concentration was lower than 104 colony-forming units (CFU)/mL. Importantly, the antibody dosage was reduced by 10 times of that in our previous study, which used antibody direct-conjugated magnetic nanoparticles. Propidium monoazide (PMA) treatment prior to PCR amplification could eliminate the false-positive results from dead bacteria and detected viable L. monocytogenes sensitively and specifically. For viable L.monocytogenes detection, enriched L. monocytogenes was treated with PMA prior to asymmetric PCR amplification. The detection limits of the combined IMS with nucleic acid lateral flow (NALF) biosensor for viable L. monocytogenes detection were 3.5 × 103 CFU/mL in phosphate buffer solution and 3.5 × 104 CFU/g in lettuce samples. The whole assay process of recognizing viable L. monocytogenes was completed within 6 h. The proposed biotin-exposure-mediated IMS combined with a disposable NALF biosensor platform posed no health risk to the end user, and possessed potential applications in the rapid screening and identification of foodborne pathogens.

Developments in Micro- and Nanotechnology for Foodborne Pathogen Detection.

In response to the potential hazards associated with the globalization of the food industry, research has been focused on the development of new sensing techniques to provide the means of contamination detection at any stage in the food supply chain. The demand for on-site detection is growing as pre-emptive sensing of pathogens could eliminate foodborne-related outbreaks and associated healthcare costs. Reduction in food waste is also a driver for point-of-use (POU) sensing, from both an economic and environmental standpoint. The following review discusses the latest advancements in platforms that have the greatest potential for inexpensive, real-time detection, and identification of foodborne pathogens. Specific focus has been placed on the development techniques, which utilize micro- and nanoscale technology. Sample preparation-free techniques are also discussed, as the growing demand to enable POU sensing at any stage in the food supply chain will be a major driver toward the advancements of these nondestructive methods.

Foodborne bacteria in dairy products: Detection by molecular techniques

Because of their unique composition and properties, milk and dairy products represent excellent growth media for many pathogenic microorganisms. Staphylococcus aureus , Salmonella spp. , Listeria monocytogenes and Escherichia coli O157:H7 are the most frequent potential pathogens associated with milk or dairy products in industrialized countries and are therefore the main microbiological hazards linked to raw milk and raw cheese. This review summarizes the scientific information about outbreaks related to foodborne pathogens in dairy products and highlights the increasing application of molecular approaches to detect and identify the bacteria responsible for these outbreaks. Molecular techniques have facilitated the rapid detection and identification of foodborne pathogens, which has been crucial for current surveillance and outbreak control.

Advanced molecular diagnostic techniques for detection of food-borne pathogens: Current applications and future challenges

ABSTRACTThe elimination of disease-causing microbes from the food supply is a primary goal and this review deals with the overall techniques available for detection of food-borne pathogens. Now-a-days conventional methods are replaced by advanced methods like Biosensors, Nucleic Acid-based Tests (NAT), and different PCR-based techniques used in molecular biology to identify specific pathogens. Bacillus cereus, Staphylococcus aureus, Proteus vulgaris, Escherichia coli, Campylobacter, Listeria monocytogenes, Salmonella spp., Aspergillus spp., Fusarium spp., Penicillium spp., and pathogens are detected in contaminated food items that cause always diseases in human in any one or the other way. Identification of food-borne pathogens in a short period of time is still a challenge to the scientific field in general and food technology in particular. The low level of food contamination by major pathogens requires specific sensitive detection platforms and the present area of hot research looking forward to new nanomolecular techniques for nanomaterials, make them suitable for the development of assays with high sensitivity, response time, and portability. With the sound of these, we attempt to highlight a comprehensive overview about food-borne pathogen detection by rapid, sensitive, accurate, and cost affordable in situ analytical methods from conventional methods to recent molecular approaches for advanced food and microbiology research.

Rapid and visual detection of Listeria monocytogenes based on nanoparticle cluster catalyzed signal amplification

Foodborne pathogens pose a significant threat to human health worldwide. The identification of foodborne pathogens needs to be rapid, accurate and convenient. Here, we constructed a nanoparticle cluster (NPC) catalyzed signal amplification biosensor for foodborne pathogens visual detection. In this work, vancomycin (Van), a glycopeptide antibiotic for Gram-positive bacteria, was used as the first molecular recognition agent to capture Listeria monocytogenes (L. monocytogenes). Fe3O4 NPC modified aptamer, was used as the signal amplification nanoprobe, specifically recognize to the cell wall of L. monocytogenes. As vancomycin and aptamer recognize L. monocytogenes at different sites, the sandwich recognition showed satisfied specificity. Compared to individual Fe3O4 nanoparticle (NP), NPC exhibit collective effect-enhanced catalytic activity for the color reaction of chromogenic substrate. The change in absorbance or color could represent the concentration of target. Using the Fe3O4 NPC-based signal amplification method, L. monocytogenes whole cells could be directly assayed within a linear range of 5.4×103–108 cfu/mL and a visual limit of detection of 5.4×103 cfu/mL. Fe3O4 NPC-based method was more sensitive than the Fe3O4 NP-based method. All these attractive characteristics of highly sensitivity, visual and labor-saving, make the biosensor possess a potential application for foodborne pathogenic bacteria detection.

Técnicas moleculares para la detección e identificación de patógenos en alimentos: ventajas y limitaciones

Foodborne diseases, caused by pathogenic microorganisms, are a major public health problem worldwide. Microbiological methods commonly used in the detection of these foodborne pathogens are laborious and time consuming. This situation, coupled with the demand for immediate results and with technological advances, has led to the development of a wide range of rapid methods in recent decades. On this basis, this review describes the advantages and limitations of the main molecular methods used in detection and identification of foodborne pathogens. To this end, we considered how recent the information was published, the objective analysis of the topic and its scope. Recent literature reports a significant number of alternative, sensitive and selective molecular techniques for detection, enumeration and identification of pathogenic microorganisms in food. Polymerase chain reaction (PCR) is the most popular platform, while high performance sequencing is emerging as a technique of wide applicability for the future. However, even with all the advantages of these new methodologies, their limitations should not be overlooked. For example, molecular methods are not standardized protocols, which hinders its use in some cases. For this reason, hard work should be done to overcome these limitations and improve the application of these techniques in complex matrices such as food systems.

Development of a Novel Hexa-plex PCR Method for Identification and Serotyping of Salmonella Species

Salmonella is one of the most important foodborne pathogens, which causes a huge economic burden worldwide. To detect Salmonella rapidly is very meaningful in preventing salmonellosis and decreasing economic losses. Currently, isolation of Salmonella is confirmed by biochemical and serobased serotyping methods, which are time consuming, labor intensive, and complicated. To solve this problem, a hexa-plex polymerase chain reaction (PCR) method was developed using comparative genomics analysis and multiplex PCR technology to detect Salmonella and Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Agona, Salmonella Choleraesuis, and Salmonella Pullorum simultaneously. The accuracy of this method was tested by a collection of 142 Salmonella. Furthermore, the strategy described in this article to mine serovar-specific fragments for Salmonella could be used to find specific fragments for other Salmonella serotypes and bacteria. The combination of this strategy and multiplex PCR is promising in the rapid identification of foodborne pathogens.

Development of a Magnetic Nanoparticles Microarray for Simultaneous and Simple Detection of Foodborne Pathogens

Foodborne diseases are a widespread and growing public health problem affecting both developed and developing countries, microbiologically contaminated food and water are the major causes of diarrhoeal diseases. Methods based on polymerase chain reaction (PCR) and microarrays are rapid and sensitive enough to detect very small quantities of microorganisms, however, the requirement for expensive equipments limits their application. In the present paper, we describe a method based on multiplex PCR and magnetic nanoparticles labelling for simultaneous detection of four major foodborne pathogens, including Escherichia coli O157:H7, Salmonella enterica, Vibrio cholera and Campylobacter jejuni. The process utilizes an oligonucleotide array onto which 5' biotinylated single strand PCR products were hybridized and visualized with streptavidin-coated magnetic nanoparticles (SA-MNPs), the signal from which could be detected by the naked eye, microscope or CCD camera. By employing SA-MNPs as visible labels, the microarray unambiguously distinguished all 4 pathogens with detection sensitivity up to 316 CFU/mL. Due to its high sensitivity, specificity and simple detection procedure, the magnetic bead assay provides a powerful tool for the detection and identification of foodborne pathogens in a modestly equipped laboratory.