Detection Of Foodborne Pathogens Research Articles

Induction into viable but non culturable state and outgrowth heterogeneity of Listeria monocytogenes is affected by stress history and type of growth

Exposure to sublethal stresses related to food-processing may induce a heterogenous mixture of cells that co-exist, comprising healthy, sublethally injured, dormant and dead cells. Heterogeneity in survival capacity and dormancy of single cells may impede the detection of foodborne pathogens. In this study, we exposed Listeria monocytogenes Scott A strain, to peracetic acid (PAA; 20–40 ppm) and to acidic conditions (hydrochloric (HCl) and acetic (AA) acid, adjusted to pH 2.7–3.0, to evaluate the resuscitation capacity and outgrowth kinetics of metabolically active cells in two different media. Injury and the viable-but-non-culturable (VBNC) status of cells were assessed by flow cytometry using CFDA (metabolically active) and PI (dead) staining. Stressed CFDA+PI− cells were sorted on Tryptic Soy (TS) Agar or in TS broth, both supplemented with 0.6 % Yeast Extract (TSAYE or TSBYE), to evaluate culturability. Resuscitation capacity of CFDA+PI−sorted cells (10 events/well) was monitored by visual inspection on TSAYE and by optical density measurement in TSBYE for 5 days. Sorting of L. monocytogenes viable cells (CFDA+PI−) in Ringer's solution on TSAYE and TSBYE showed 100 % recovery in both media (control condition), while the mean lag time in TSBYE was 9.6 h. Treatment with 20 ppm PAA for 90 and 180 min resulted in 74.79 % and 85.82 % of non-culturable cells in TSBYE and increased the average lag time to 41.7 h and 43.8 h, respectively, compared to the control (9.6 h). The longest average lag time (79.5 h) was detected after treatment with 30 ppm PAA for 90 min, while at the same condition sorting of CFDA+PI− cells resulted in 95.05 % and 93.94 % non-culturable cells on TSAYE and TSBYE, respectively. The highest percentage of wells with non-culturable cells (96.17 %) was detected on TSAYE after treatment with 40 ppm PAA for 30 min. Fractions of VBNC cells were detected in TSBYE after treatment with HCl pH 3.0 for 60 and 240 min, and in TSAYE and TSBYE after exposure to AA pH 2.7. Treatment with AA pH 2.7 for 150–300 min increased the range of recorded lag time values compared to 60 min, from 8.6 h up to 13.3 h, as well as the mean lag times in TSBYE. Modelling of the outgrowth kinetics comparing the two types of stress (oxidative vs acid) and the two systems of growth (colonial vs planktonic) revealed that low starting concentrations hindered the detection of viable L. monocytogenes cells, either due to VBNC induction or cell heterogeneity.

Rapid and accurate detection of foodborne pathogens: impedimetric immunosensor at the forefront

Microorganisms can be categorized into five categories: bacteria, algae, fungi, viruses, and protozoa which can cause microbiological hazards, contaminating food during production, manufacture, transportation, and storage. Biosensor technology is one of the most reliable and effective analytical technique for determining a biological system's sensitivity and specificity at a very low scale. So many techniques exist for identifying and detecting food-borne illness causing bacteria. But biosensor technology is used to detect and monitor with accuracy, thus increasing its interest globally with time. Biosensors have been used for a long time to illustrate the process of regulating data in the pharmaceutical, food manufacturing, and processing industries. An immunosensor is a type of biosensor that uses the molecular recognition specificity of antigens to form a stable complex of an antibody. It is divided into two subcategories- labeled and label-free. A labeled immunosensor is primarily used to sense the immune reaction and generate signals that allow versatile detection of complex, and detect the microorganisms like Listeria monocytogenes, Salmonella spp., and Escherichia coli. Label-free immunosensors detect physical changes during the immune complex formation and have been explored because of their potential as a specific detection technique which can reduce the time and cost of analysis. It has been developed using several detection methods like optical changes for Salmonella sp., electrochemical changes for Listeria monocytogenes and Escherichia coli, and Hepatitis B. An impedimetric immunosensor for pathogens has been created utilizing a biosensor, electrochemical impedance spectroscopy, antibodies, affinity proteins, affimers, and other binding proteins like bio-receptors, which exhibit good selectivity. This review discusses the techniques used by immunosensors to recognize microorganisms that might cause food poisoning. These techniques consider the electrodes and base-layer components, the makeup and characteristics of distinct bacterial species, as well as the interactions between antigens and antibodies that are utilized to detect bacteria.

Cured or fresh? Between fish maturation trends in restaurants and food safety: The case of dry-aged rainbow trout

The application of dry aging on fish is increasing in restaurants worldwide. This process determines fluid loss, changes in muscle texture, and the development of a more concentrated flavour appreciated by consumers. Despite the growing popularity in the foodservice industry, there is a lack of data on potential risks and quality conditions of dry-aged fish. This study was aimed at assessing the safety for consumption of whole rainbow trout subjected to dry aging in a local restaurant (Turin, Italy). Quantitative microbiological analyses and detection of foodborne pathogens were performed on dorsal, ventral muscles, and skin samples at day 0, 3, 6, 10, and 14 of dry aging. Changes in water activity (aw), pH, colour, and biogenic amines in muscle were checked at each sampling point. Almost all the microbiological indicators were below the quantification limits in muscle until day 6 of dry aging, while higher but still acceptable loads were observed at day 10. At day 14, yeasts and moulds exceeded the acceptable level in one case (3.3 ± 0.4 log10 CFU/g) in ventral muscle and counts of Pseudomonas spp. (from 6.1 ± 0.8 to 6.4 ± 0.5 log10 CFU/g), total mesophilic (from 4.4 ± 0.3 to 5.3 ± 0.7 log10 CFU/g), and psychrophilic (from 5.4 ± 0.1 to 5.8 ± 0.3 log10 CFU/g) bacteria indicated the approaching of spoilage. Detection of Listeria monocytogenes and Staphylococcus aureus was always negative during the trials. Minimal changes of pH and colour were observed, while aw values followed a decreasing trend during dry aging. Putrescine was the only detectable amine, with a maximum concentration of 2.05 ± 0.02 mg/kg at day 14. In this study, dry-aged rainbow trout was safe for consumption and 10 days was the most appropriate curing time with 3 °C and 78% of relative humidity. Further studies at different environmental conditions and on other fish species are needed for a comprehensive risk assessment of this fast-growing practice in the restaurant sector.

D-Glucose-Mediated Gold Nanoparticle Fabrication for Colorimetric Detection of Foodborne Pathogens.

Gold nanoparticle (AuNP) fabrication via the oxidation of D-glucose is applied for detecting two foodborne pathogens, Enterococcus faecium (E. faecium) and Staphylococcus aureus (S. aureus). D-glucose is used as a reducing agent due to its oxidation to gluconic acid by sodium hydroxide (NaOH), resulting in the formation of AuNPs. Based on this mechanism, we develop AuNP-based colorimetric detection in conjunction with loop-mediated isothermal amplification (LAMP) for accurately identifying the infectious bacteria. Here, Au+ ions bind to the base of double-stranded DNA. In the presence of D-glucose and NaOH, the LAMP amplicon-Au+ complex maintains its bound state at 65 °C for 10 min while it is reduced to AuNPs in a dispersed form, exhibiting a red color. We aimed to pre-mix D-glucose with LAMP reagents before amplification and induce successful colorimetry without inhibiting amplification to simplify the experimental process and decrease the reaction time. Therefore, the entire process, including LAMP and colorimetric detection, is accomplished in approximately 1 h. The limit of detection of E. faecium and S. aureus is confirmed using the introduced method as 101 CFU/mL and 100 fg/μL, respectively. We expect that colorimetric detection using D-glucose-mediated AuNP synthesis offers an application for simple and immediate molecular diagnosis.

3D printed microfluidic chip integrated with nanointerferometer for multiplex detection of foodborne pathogens

The current foodborne pathogen detection methods, such as culture-based methods, polymerase chain reaction, and optical and electrochemical biosensors with nucleic acid, have high sensitivity and selectivity. However, they are slow, expensive, and require well-trained operators. In this study, we utilized a 3D printer to develop a novel chip with an aptamer-based nanointerferometer capable of identifying four distinct foodborne pathogens: Listeria monocytogenes, Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus. The aptamer sensor on the chip achieved a limit-of-detection of 10 colony forming unit (CFU)/ml. With its high sensitivity and specificity, this chip offers a cost-effective platform for distinguishing and screening different foodborne pathogens.

Gold Nanoparticle-Based Plasmonic Detection of Escherichia coli, Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes from Bovine Fecal Samples.

Current diagnostic methods for detecting foodborne pathogens are time-consuming, require sophisticated equipment, and have a low specificity and sensitivity. Magnetic nanoparticles (MNPs) and plasmonic/colorimetric biosensors like gold nanoparticles (GNPs) are cost-effective, high-throughput, precise, and rapid. This study aimed to validate the use of MNPs and GNPs for the early detection of Escherichia coli O157:H7, Salmonella enterica spp., Campylobacter jejuni, and Listeria monocytogenes in bovine fecal samples. The capture efficiency (CE) of the MNPs was determined by using Salmonella Typhimurium (ATCC_13311) adjusted at an original concentration of 1.5 × 108 CFU/mL. One (1) mL of this bacterial suspension was spiked into bovine fecal suspension (1 g of fecal sample in 9 mL PBS) and serially diluted ten-fold. DNA was extracted from Salmonella Typhimurium to determine the analytical specificity and sensitivity/LOD of the GNPs. The results showed that the CE of the MNPs ranged from 99% to 100% and could capture as little as 1 CFU/mL. The LOD of the GNPs biosensor was 2.9 µg/µL. The GNPs biosensor was also tested on DNA from 38 naturally obtained bovine fecal samples. Out of the 38 fecal samples tested, 81.6% (31/38) were positive for Salmonella enterica spp., 65.8% (25/38) for C. jejuni, 55.3% (21/38) for L. monocytogenes, and 50% (19/38) for E. coli O157:H7. We have demonstrated that MNP and GNP biosensors can detect pathogens or their DNA at low concentrations. Ensuring food safety throughout the supply chain is paramount, given that these pathogens may be present in cattle feces and contaminate beef during slaughter.

Evaluating Commercial Loop-Mediated Isothermal Amplification Master Mixes for Enhanced Detection of Foodborne Pathogens.

Loop-mediated isothermal amplification, LAMP, is nowadays the most popular isothermal nucleic acid amplification technique, and as such, several commercial, ready-to-use master mixes have flourished. Unfortunately, independent studies to determine their performance are limited. The current study performed an independent evaluation of the existing ready-to-use commercial LAMP master mixes WarmStart® LAMP Kit, LavaLAMP™ DNA Master Mix, Saphir Bst Turbo GreenMaster, OptiGene Fast Master Mix ISO-004, and SynLAMP Mix. To reduce bias, three different genes, namely ttr (Salmonella spp.), rfbE (E. coli O157), and hly (Listeria monocytogenes), were targeted. The comparison was based on amplification speed, performance with decreasing DNA concentrations, and the effect of five typical LAMP reaction additives (betaine, DMSO, pullulan, TMAC, and GuHCl). Significant differences were observed among the different master mixes. OptiGene provided the fastest amplification and showed less detrimental effects associated with the supplements evaluated. Out of the chemicals tested, pullulan provided the best results in terms of amplification speed. It is noteworthy that the different additives impacted the master mixes differently. Overall, the current study provides insights into the performance of commercial LAMP master mixes, which can be of value for the scientific community to better select appropriate reagents when developing new methods.

Flexible Biosensors for Food Pathogen Detection

AbstractFood contamination poses a significant threat to public health, the economy, and human health worldwide, occurring at any stage of the food supply chain, from farm to fork. Efficient and effective real‐time monitoring methods for the early identification and rapid detection of pathogen contamination are critical to preventing possible food safety issues. In the past decade, flexible electrochemical biosensors have rapidly expanded in the detection of foodborne pathogens, owing to their ability to function well at biological interfaces that may be soft, intrinsically curvy, irregular, or deformable. The most important features of flexible sensors are their flexibility, multifunctionality, low cost, and good compatibility with solution processing methods. These properties enable flexible sensors to be used in various applications, including smart packaging. This review focuses on the materials, structures, and fabrication methods of flexible biosensors, as well as their applications in food safety monitoring. It examines recent advances in electrochemical immunosensors and aptasensors for pathogen detection, with a focus on their key components and their application in food analysis, while critically examining their current trends, particularly in conjunction with nanomaterials such as carbon nanotubes, graphene, gold nanoparticles, Indium tin oxide, etc. Finally, current issues and prospects for the future are discussed.

Research progress on the detection of foodborne pathogens based on aptamer recognition.

Foodborne diseases caused by bacterial contamination are a serious threat to food safety and human health. The classical plate culture method has the problems of long detection cycle, low sensitivity and specificity, and complicated operation, which cannot meet the growing demand for rapid quantitative detection of pathogenic bacteria. The frequent outbreak of foodborne diseases has put forward higher requirements for rapid and simple detection technology of foodborne pathogens. Aptamer is a kind of oligonucleotide fragment that can recognize targets with the advantages of high affinity and good specificity. The target can be range from proteins, small molecules, cells bacteria, and even viruses. Herein, the latest advances in sensitive and rapid detection of foodborne pathogens based on aptamer recognition was reviewed. Special attention has been paid to the obtained sequences of aptamers to various foodborne pathogens, the optimization of sequences, and the mechanism of aptamer recognition. Then, the research progress of biosensors for the detection of pathogenic bacteria based on aptamer recognition were summarized. Some challenges and prospects for the detection of foodborne pathogens based on aptamer recognition were prospected. In summary, with the further deepening of aptamer research and improvement of detection technology, aptamer-based recognition can meet the needs of rapid, sensitive, and accurate detection in practical applications.

Comparing two magnetic separation pretreatment methods for molecular detection using (q)PCR assays targeting Staphylococcus aureus in milk as models

Magnetic separation presents significant potential for culture-independent detection of foodborne pathogens in food samples. In this study, we compared two magnetic separation pretreatment strategies for molecular detection using (q)PCR assays targeting Staphylococcus aureus in milk as models. The first approach employed amino-modified silica magnetic particles (ASMPs) for DNA separation, while the second utilized pig IgG-labeled magnetic beads (IgG-MBs) for cell separation. In the ASMPs-based DNA separation strategy, a sensitivity of 36 CFU/mL was achieved when ASMPs-DNA complexes were employed as templates for PCR analysis. To mitigate noise signals originating from ASMPs during qPCR, DNA on the surface of ASMPs was replaced with dNTP, resulting in a sensitivity of 1.6 × 103 CFU/mL. The IgG-MBs-based cell separation approach yielded sensitivities of 3.6 × 104 CFU/mL for PCR and 1.6 × 103 CFU/mL for qPCR following DNA isolation from the bacteria-IgG-MBs complexes. Both methods exhibited exceptional specificity and robustness against background bacteria interference. However, neither approach effectively discriminated between live and dead bacteria. In comparison, the ASMPs-based DNA separation strategy exhibited superior potential especially when ASMPs do not influence the detection system. This research contributes to the optimization of magnetic separation pretreatment strategies for the molecular detection of foodborne pathogens, highlighting the potential of culture-independent detection methods.

WO3 Nanoplates Decorated with Au and SnO2 Nanoparticles for Real-Time Detection of Foodborne Pathogens.

Nowadays, metal oxide semiconductor gas sensors have diverse applications ranging from human health to smart agriculture with the development of Internet of Things (IoT) technologies. However, high operating temperatures and an unsatisfactory detection capability (high sensitivity, fast response/recovery speed, etc.) hinder their integration into the IoT. Herein, a ternary heterostructure was prepared by decorating WO3 nanoplates with Au and SnO2 nanoparticles through a facial photochemical deposition method. This was employed as a sensing material for 3-hydroxy-2-butanone (3H-2B), a biomarker of Listeria monocytogenes. These Au/SnO2-WO3 nanoplate-based sensors exhibited an excellent response (Ra/Rg = 662) to 25 ppm 3H-2B, which was 24 times higher than that of pure WO3 nanoplates at 140 °C. Moreover, the 3H-2B sensor showed an ultrafast response and recovery speed to 25 ppm 3H-2B as well as high selectivity. These excellent sensing performances could be attributed to the rich Au/SnO2-WO3 active interfaces and the excellent transport of carriers in nanoplates. Furthermore, a wireless portable gas sensor equipped with the Au/SnO2-WO3 nanoplates was assembled, which was tested using 3H-2B with known concentrations to study the possibilities of real-time gas monitoring in food quality and safety.

One-pot dual-CRISPR/Cas12b-mediated dUTP-LAMP for rapid and sensitive detection of foodborne pathogens without carryover contamination

Rapid and accurate detection of foodborne pathogens is of great significance for ensuring food safety. Currently, the CRISPR-Dx based on Cas12b, a new and better performing Cas enzyme, is very rare, and the vast majority require a two-step method, facing aerosol contamination. In this work, we reported a one-pot dual-CRISPR/Cas12b mediated dUTP-LAMP method for rapid and sensitive detection of foodborne pathogens without nucleic acid extraction and contamination-free. The design mechanism of sgRNAs in the one-pot CRISPR/Cas12b-mediated LAMP method was investigated in detail for the first time. Moreover, a novel PAM sequence UUN of AapCas12b was discovered, and combined with dUTP-LAMP system for one-pot dual-sgRNA detection without carryover contamination. Our developed method can achieve high specificity, nucleic acid extraction-free detection of 2.33 × 104 CFU/mL Salmonella within 1 h. Furthermore, accurate detection of Salmonella was successfully performed in real samples of milk, fruit and egg, demonstrating the practical significance of the developed method.

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

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

A CRISPR-Cas12a-powered, quantum dot-based and magnetic nanoparticle-assisted (QD-CRISPR-MNP) biosensor for the screening of Salmonella

Rapid detection methods are urgently needed to help prevent the spread of Salmonella among food products. Herein, a CRISPR-Cas12a-powered, quantum dot-based and magnetic nanoparticle-assisted (QD-CRISPR-MNP) biosensor was proposed for the screening of Salmonella. The QD-CRISPR-MNP biosensor was designed and fabricated relying on the CRISPR-Cas12a system for better guidance of the coupling between magnetic nanoparticles (MNPs) and quantum dots (QDs). In this biosensor, the activity of the CRISPR-Cas12a system was unlocked by Salmonella and displayed collateral cleavage activity towards linker DNAs, which inhibited the coupling of the MNPs and QDs. Then, Salmonella could be quantitatively detected by simply measuring the fluorescence emitted by the QDs on the MNPs surface. This biosensor showed good detection performance for Salmonella with a detection limit of 86 CFU mL−1, and exhibited a satisfactory specificity as well. Moreover, it overcomes the low fluorescence quantum yields and the possible high fluorescence background noise in conventional dye-based CRISPR detection platforms, acting as a powerful alternative for the detection of foodborne pathogens.

Photothermal-Based Multiplex Nested Digital PCR System for Rapid Detection of Foodborne Pathogens.

The rapid and sensitive detection of foodborne pathogens is crucial for ensuring food safety. Among virus testing methods, polymerase chain reaction (PCR) has served as the gold-standard technique in most food safety regulation organizations. However, to enhance the speed and efficiency of PCR, novel approaches are continually being explored. In this work, leveraging the photothermal effects and high thermal conductivity of gold nanoparticles, we have significantly improved the heating and cooling rates of thermal cycles, enabling ultra-fast PCR detection. Specifically, we present a pre-degassing multiplex digital PCR chip integrated with gold nanoparticles. We further developed a portable system with a light source for photothermal heating cycling, along with an optoelectronic sensor to analyze PCR amplification products after rapid thermal cycling. As proof of concept, the proposed chip and portable device was applied for the on-site detection of several types of foodborne pathogens, including Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, and Salmonella. The whole system could distinguish those pathogens within 20 min, showing good potential for the rapid detection of multiple types of foodborne pathogens.

Sensitive colorimetric aptasensor for multiple foodborne pathogens detection based on PCN-Mo peroxidase-like activity

A rapid and sensitive colorimetric biosensor was developed for simultaneous detection of various pathogenic bacteria based on aptamer-mediated peroxidase (POD)-like activity of PCN-Mo nanozyme. Initially, a porphyrin-based PCN-Mo metal organic framework (MOF) nanozyme is constructed through a simple hydrothermal method, as thoroughly figured out by a series of characterization techniques. The study showed that the pathogenic bacteria aptamers could adhere to the surface of PCN-Mo, and significantly enhanced the POD-like activity by reinforcing their transport of electrons to 3,3′,5,5′-tetramethylbenzidine (TMB). In the presence of different target bacteria, the aptamers tend to bind to bacteria specifically, causing the further enhancement of POD-like activity. Based on the signal-amplification by the PCN-Mo@aptamer (PCN-Mo@Apt) nanozyme, the constructed pathogenic bacteria sensor demonstrated a wide linear range between 101 to 105 CFU/mL, with the detection limits (LODs) of 0.578, 0.439, and 0.643 CFU/mL for E. coli, S. typhimurium and S. aureus, respectively. Furthermore, the PCN-Mo sensor was capable of detection pathogenic bacteria in water, milk, classic milk tea, chicken, and beef with the recoveries of 90.1%–104.1%, indicating the sensor's practicability for pathogenic bacteria detection in real samples.

Mesoporous Silica Nanoparticles-Enhanced Microarray Technology for Highly Sensitive Simultaneous Detection of Multiplex Foodborne Pathogens.

Ensuring food safety is paramount for the food industry and global health concerns. In this study, we have developed a method for the detection of prevalent foodborne pathogenic bacteria, including Escherichia coli, Salmonella spp., Listeria spp., Shigella spp., Campylobacter spp., Clostridium spp., and Vibrio spp., utilizing antibody-aptamer arrays. To enhance the fluorescence signals on the microarray, the mesoporous silica nanoparticles (MSNs) conjugated with fluorescein, streptavidin, and seven detection antibodies-biotin were employed, forming fluorescein doped mesoporous silica nanoparticles conjugated with detection antibodies (MSNs-Flu-SA-Abs) complexes. The array pattern was designed for easy readability and enabled the simultaneous detection of all seven foodborne pathogens, referred to as the 7FP-biochip. Following the optimization of MSNs-Flu-SA-Abs complexes attachment and enhancement of the detection signal in fluorescent immunoassays, a high level of sensitivity was achieved. The detection limits for the seven pathogens in both buffer and food samples were 102 CFU/mL through visual screening, with fluorescent intensity quantification achieving levels as low as 20-34 CFU/g were achieved on the antibody-aptamer arrays. Our antibody-aptamer array offers several advantages, including significantly reduced nonspecific binding with no cross-reaction between bacteria. Importantly, our platform detection exhibited no cross-reactivity among the tested bacteria in this study. The multiplex detection of foodborne pathogens in canned tuna samples with spiked bacteria was successfully demonstrated in real food measurements. In conclusion, our study presents a promising method for detecting multiple foodborne pathogens simultaneously. With its high sensitivity and specificity, the developed antibody-aptamer array holds great potential for enhancing food safety and public health.

Development of a 3D Microfluidic Analytical Device for the Detection of Pathogenic Bacteria in Commercial Food Samples with Loop-Mediated Isothermal Amplification

Traditional methods of detecting foodborne pathogens take several days to produce the required results. Furthermore, various molecular techniques (e.g., PCR) that also produce reliable results in the detection of pathogenic bacteria have been introduced, but the cost–time ratio required does not allow them to be considered a substantial solution to this specific problem. Three-dimensional (3D) printing technology provides the ability to design and manufacture microfluidic analytical devices using conventional 3D printers, which, in combination with colorimetric loop-mediated isothermal amplification (LAMP), may further simplify the process. The overall reduction in time and cost may provide the opportunity to upscale this diagnostic modality. Moreover, unlike most microfluidic analytical devices, this technique is simpler and more user-friendly, as it does not require any expertise or additional equipment apart from a conventional oven. A 3D-printed microfluidic analytical device in combination with LAMP was developed and tested for the simultaneous detection of foodborne pathogens in food samples. A total of 150 commercial food specimens (50 milk, 50 chicken, 50 lettuce samples) were analyzed for possible contamination with Salmonella typhimurium, Listeria monocytogenes and Escherichia coli. The 3D-printed microfluidic device was 100% precise for both negative (80 samples) and positive samples (7 samples were positive for S. typhimurium, 28 for L. monocytogenes, and 35 for E. coli) for all pathogens. Overall, the amount of data analyzed led to a high level of confidence in the precision of this device. As such, this new 3D device in combination with LAMP provides a precise detection method for food pathogens with a low detection limit.

Bacteriophage-assisted lysis and eluted genomic DNA-based detection of pathogenic bacterial contamination in food

Foodborne diseases represent a significant global health concern, with a substantial number of illnesses and fatalities reported annually owing to the consumption of contaminated food. Current methods for identifying foodborne pathogens cannot effectively distinguish live from dead bacteria or efficiently detect multiple targets. Therefore, we developed a selective detection method for three pathogenic bacteria in food samples that leverages bacteriophage-assisted live-cell lysis and exposed genomic DNA amplification. The antimicrobial activity and host specificity of these three bacteriophages were employed to confirm the target bacteria selectivity. Bacterial DNA purification using bacteriophage-assisted lysis demonstrated the superior activity of the selected phages in extracting DNA from Escherichia coli O157:H7 and Salmonella Typhimurium. The results from reactions involving the three strains, including Bacillus cereus, highlighted that a 1 h incubation period yielded the most efficient DNA extraction and purification. The polymerase chain reaction (PCR) products obtained were comparable in terms of concentration and purity with those generated using bacterial DNA prepared using a commercial kit. In the practical food sample application experiment, bacterial strains were successfully recovered and concentrated using a polyethersulfone filter, resulting in a concentration factor >3 for all samples. Furthermore, the concentration of purified bacterial DNA from contaminated kimchi cabbage exceeded 10 ng/μL. Notably, we could detect E. coli O157:H7, S. Typhimurium, and B. cereus spiked at 103 colony-forming units/mL using multiplex PCR. This innovative bacteriophage-based approach holds substantial promise for the selective and practical detection of live foodborne pathogens, thereby significantly advancing food safety standards.

Dynamically reconfigurable complex liquid crystal emulsions

ABSTRACT Complex emulsions, containing liquid crystals (LCs) and immiscible oils such as fluorocarbon or silicone oils, present innovative functionalities in soft materials. These emulsions exhibit dynamic reconfiguration in droplet morphology and internal LC organisation, expanding applications in optics, sensing, and active matter. This review highlights the evolution and potential of LC-containing complex emulsions. Emphasis is placed on emulsions featuring dynamic and reversible morphological transformations and LC reorganisation, underpinning their suitability for tunable micro-lenses, chemical sensing, and biosensing. Nematic LCs within complex emulsions allow for topological defect-driven functionalization, attaching antibodies to induce substantial changes in the LC director field upon interfacial recognition events. This feature may facilitate the development of ultrasensitive chemical and biological sensors. Moreover, cholesteric LCs, through their selective reflection, enable the rapid and sensitive detection of foodborne pathogens. The controlled assembly of magnetic nanoparticles at interfaces illustrates advancements towards magnetic field-responsive and active colloids. Future perspectives include exploiting these properties for intelligent colloidal systems capable of autonomous behaviour, and furthering applications in dynamic photonic devices, high-security identification tags, and responsive lens systems.