Rapid Detection and Identification of Vancomycin-Sensitive Bacteria Using an Electrochemical Apta-Sensor.

In order to combat the complex and diverse infections caused by bacteria, it is essential to develop efficient diagnostic tools. Current techniques for bacterial detection rely on laborious multistep procedures, with high costs and extended time of analysis. To overcome these limitations, we propose here a novel portable electrochemical biosensor for the rapid detection and identification of Gram-positive bacteria that leverages the recognition capabilities of vancomycin and aptamers. A vancomycin-modified screen-printed carbon electrode was used to selectively capture Gram-positive bacteria susceptible to this antibiotic. Electrochemical impedance spectroscopy and scanning electron microscopy demonstrated that capture was achieved in 10 min, with a limit of detection of only 2 CFU/mL. We then tested the device's potential for aptamer-based bacterial identification using Staphylococcus aureus and Bacillus cereus as the test strains. Specifically, electrodes with captured bacteria were exposed to species-specific aptamers, and the resulting changes in current intensity were analyzed using differential pulse voltammetry. When used directly in untreated milk or serum, the system was able to successfully identify a small amount of S. aureus and B. cereus (100 CFU/mL) in less than 45 min. This novel biosensor has the potential to serve as an invaluable tool that could be used, even by inexperienced staff, in a broad range of settings including clinical diagnostics, food safety analysis, environmental monitoring, and security applications.

Luminescent Metal-Organic Frameworks for Electrochemiluminescent Detection of Water Pollutants.

The modern lifestyle has increased our utilization of pollutants such as heavy metals, aromatic compounds, and contaminants that are of rising concern, involving pharmaceutical and personal products and other materials that may have an important environmental impact. In particular, the ultimate results of the intense use of highly stable materials, such as heavy metals and chemical restudies, are that they turn into waste materials, which, when discharged, accumulate in environmental water bodies. In this context, the present review presents the application of metal-organic frameworks (MOFs) in electrochemiluminescent (ECL) sensing for water pollutant detection. MOF composites applied as innovative luminophore or luminophore carriers, materials for electrode modification, and the enhancement of co-reaction in ECL sensors have enabled the sensitive monitoring of some of the most common contaminants of emerging concern such as heavy metals, volatile organic compounds, pharmaceuticals, industrial chemicals, and cyanotoxins. Moreover, we provide future trends and prospects associated with ECL MOF composites for environmental sensing.

Comparative meta-analysis of antimicrobial resistance from different food sources along with one health approach in the Egypt and UK.

BACKGROUND: Antimicrobial resistance (AMR) is a critical global issue that poses significant threats to human health, animal welfare, and the environment. With the increasing emergence of resistant microorganisms, the effectiveness of current antimicrobial medicines against common infections is diminishing. This study aims to conduct a competitive meta-analysis of surveillance data on resistant microorganisms and their antimicrobial resistance patterns in two countries, Egypt and the United Kingdom (UK). METHODS: Data for this study were obtained from published reports spanning the period from 2013 to 2022. In Egypt and the UK, a total of 9,751 and 10,602 food samples were analyzed, respectively. Among these samples, 3,205 (32.87%) in Egypt and 4,447 (41.94%) in the UK were found to contain AMR bacteria. RESULTS: In Egypt, the predominant resistance was observed against ß-lactam and aminoglycosides, while in the United Kingdom, most isolates exhibited resistance to tetracycline and ß-lactam. The findings from the analysis underscore the increasing prevalence of AMR in certain microorganisms, raising concerns about the development of multidrug resistance. CONCLUSION: This meta-analysis sheds light on the escalating AMR problem associated with certain microorganisms that pose a higher risk of multidrug resistance development. The significance of implementing One Health AMR surveillance is emphasized to bridge knowledge gaps and facilitate accurate AMR risk assessments, ensuring consumer safety. Urgent actions are needed on a global scale to combat AMR and preserve the effectiveness of antimicrobial treatments for the well-being of all living beings.

Direct glyphosate soil monitoring at the triazine-based covalent organic framework with the theoretical study of sensing principle.

Covalent organic frameworks (COFs) are emerging as promising sensing materials due to their controllable structure and function properties, as well as excellent physicochemical characteristics. Here, specific interactions between a triazine-based COF and a mass-used herbicide - glyphosate (GLY) have been utilized to design a disposable sensing platform for GLY detection. This herbicide has been extensively used for decades, however, its harmful environmental impact and toxicity to humans have been recently proven, conditioning the necessity for the strict control and monitoring of its use and its presence in soil, water, and food. Glyphosate is an organophosphorus compound, and its detection in complex matrices usually requires laborious pretreatment. Here, we developed a direct, miniaturized, robust, and green approach for disposable electrochemical sensing of glyphosate, utilizing COF's ability to selectively capture and concentrate negatively charged glyphosate molecules inside its nanopores. This process generates the concentration gradient of GLY, accelerating its diffusion towards the electrode surface. Simultaneously, specific COF-glyphosate binding catalyses the oxidative cleavage of the C-P bond and, together with pore nanoconfinement, enables sensitive glyphosate detection. Detailed sensing principles and selectiveness were scrutinized using DFT-based modelling. The proposed electrochemical method has a linear working range from 0.1 µM to 10 µM, a low limit of detection of 96 nM, and a limit of quantification of 320 nM. The elaborated sensing approach is viable for use in real sample matrices and tested for GLY determination in soil and water samples, without pretreatment, preparation, or purification. The results showed the practical usefulness of the sensor in the real sample analysis and suggested its suitability for possible out-of-laboratory sensing.

Nanobiotics and the One Health Approach: Boosting the Fight against Antimicrobial Resistance at the Nanoscale.

Antimicrobial resistance (AMR) is a growing public health concern worldwide, and it poses a significant threat to human, animal, and environmental health. The overuse and misuse of antibiotics have contributed significantly and others factors including gene mutation, bacteria living in biofilms, and enzymatic degradation/hydrolyses help in the emergence and spread of AMR, which may lead to significant economic consequences such as reduced productivity and increased health care costs. Nanotechnology offers a promising platform for addressing this challenge. Nanoparticles have unique properties that make them highly effective in combating bacterial infections by inhibiting the growth and survival of multi-drug-resistant bacteria in three areas of health: human, animal, and environmental. To conduct an economic evaluation of surveillance in this context, it is crucial to obtain an understanding of the connections to be addressed by several nations by implementing national action policies based on the One Health strategy. This review provides an overview of the progress made thus far and presents potential future directions to optimize the impact of nanobiotics on AMR.

Recent Advances in Electrochemical Biosensors for Food Control.

The rapid and sensitive detection of food contaminants is becoming increasingly important for timely prevention and treatment of foodborne disease. In this review, we discuss recent developments of electrochemical biosensors as facile, rapid, sensitive, and user-friendly analytical devices and their applications in food safety analysis, owing to the analytical characteristics of electrochemical detection and to advances in the design and production of bioreceptors (antibodies, DNA, aptamers, peptides, molecular imprinted polymers, enzymes, bacteriophages, etc.). They can offer a low limit of detection required for food contaminants such as allergens, pesticides, antibiotic traces, toxins, bacteria, etc. We provide an overview of a broad range of electrochemical biosensing designs and consider future opportunities for this technology in food control.

Colorimetric aptasensor for detection of Bacillus cytotoxicus spores in milk and ready-to-use food.

The high incidence of foodborne diseases caused by pathogenic bacteria raises concerns worldwide and imposes considerable public healthcare challenges. This is especially observed with dormant spores of Bacilli, which can often survive treatments used by the food industry to kill growing bacteria. The early and rapid detection of bacterial spores is essential to ensure food safety. Commercial availability of such a test will present a high potential for food sector. We present a point-of-need colorimetric assay for detection of Bacillus cytotoxicus spores in food. The detection principle is based on spore-enhanced peroxidase-like catalytic activity of gold nanoparticles. The sensing platform consists of a microtube containing gold nanoparticles (AuNPs), and magnetic particles (MPs), both conjugated with specific aptamer BAS6R that recognize B. cytotoxicus spores. Upon the addition of the sample, spores were determined as present by the enhanced color change of the solution, due to the oxidation of tetramethylbenidine (TMB) with H2O2. The assay was evaluated by the naked eye (on/off) and quantitatively with use of a spectrophotometer. BAS6R@AuNPs aptasensor coupled to BAS6R@MPs proved to be highly sensitive, achieving the naked-eye limit of detection as low as 102 cfu/mL in water and milk, and 104 cfu/mL in mashed potatoes. Moreover, discrimination between spores of B. cytotoxicus and B. subtilis as well as bacterial vegetative cells was achieved in contaminated food samples, providing a good selectivity. This work provides a promising proof of concept for the development of instrument-free, low-cost and rapid assay for Bacillus cytotoxicus spore detection, which is able to compete in sensitivity with conventional costly and time-consuming laboratory analyses.

The Tolerance of Gut Commensal Faecalibacterium to Oxidative Stress Is Strain Dependent and Relies on Detoxifying Enzymes.

Obligate anaerobic bacteria in genus Faecalibacterium are among the most dominant taxa in the colon of healthy individuals and contribute to intestinal homeostasis. A decline in the abundance of this genus is associated with the occurrence of various gastrointestinal disorders, including inflammatory bowel diseases. In the colon, these diseases are accompanied by an imbalance between the generation and elimination of reactive oxygen species (ROS), and oxidative stress is closely linked to disruptions in anaerobiosis. In this work, we explored the impact of oxidative stress on several strains of faecalibacteria. An in silico analysis of complete genomes of faecalibacteria revealed the presence of genes encoding O2- and/or ROS-detoxifying enzymes, including flavodiiron proteins, rubrerythrins, reverse rubrerythrins, superoxide reductases, and alkyl peroxidase. However, the presence and the number of these detoxification systems varied greatly among faecalibacteria. These results were confirmed by O2 stress survival tests, in which we found that strains differed widely in their sensitivity. We showed the protective role of cysteine, which limited the production of extracellular O2•- and improved the survival of Faecalibacterium longum L2-6 under high O2 tension. In the strain F. longum L2-6, we observed that the expression of genes encoding detoxifying enzymes was upregulated in the response to O2 or H2O2 stress but with different patterns of regulation. Based on these results, we propose a first model of the gene regulatory network involved in the response to oxidative stress in F. longum L2-6. IMPORTANCE Commensal bacteria in the genus Faecalibacterium have been proposed for use as next-generation probiotics, but efforts to cultivate and exploit the potential of these strains have been limited by their sensitivity to O2. More broadly, little is known about how commensal and health-associated bacterial species in the human microbiome respond to the oxidative stress that occurs as a result of inflammation in the colon. In this work, we provide insights regarding the genes that encode potential mechanisms of protection against O2 or ROS stress in faecalibacteria, which may facilitate future advances in work with these important bacteria.

Therapeutic applications of nanobiotechnology.

Nanobiotechnology, as a novel and more specialized branch of science, has provided a number of nanostructures such as nanoparticles, by utilizing the methods, techniques, and protocols of other branches of science. Due to the unique features and physiobiological characteristics, these nanostructures or nanocarriers have provided vast methods and therapeutic techniques, against microbial infections and cancers and for tissue regeneration, tissue engineering, and immunotherapies, and for gene therapies, through drug delivery systems. However, reduced carrying capacity, abrupt and non-targeted delivery, and solubility of therapeutic agents, can affect the therapeutic applications of these biotechnological products. In this article, we explored and discussed the prominent nanobiotechnological methods and products such as nanocarriers, highlighted the features and challenges associated with these products, and attempted to conclude if available nanostructures offer any scope of improvement or enhancement. We aimed to identify and emphasize the nanobiotechnological methods and products, with greater prospect and capacity for therapeutic improvements and enhancements. We found that novel nanocarriers and nanostructures, such as nanocomposites, micelles, hydrogels, microneedles, and artificial cells, can address the associated challenges and inherited drawbacks, with help of conjugations, sustained and stimuli-responsive release, ligand binding, and targeted delivery. We recommend that nanobiotechnology, despite having few challenges and drawbacks, offers immense opportunities that can be harnessed in delivering quality therapeutics with precision and prediction. We also recommend that, by exploring the branched domains more rigorously, bottlenecks and obstacles can also be addressed and resolved in return.

Low-cost goldleaf electrode as a platform for Escherichia coli immunodetection.

Gold electrodes are one of most prevalent substrates in electrochemical biosensors because they can be easily and highly efficiently functionalized with thiolated biomolecules. However, conventional methods to fabricate gold electrodes are costly, time-consuming and require onerous equipment. Here, an affordable method for rapid fabrication of an electrochemical immunosensor for Escherichia coli detection is presented. The gold electrode was generated using 24-karat gold leaves and lowcost polyvinyl chloride adhesive sheets covered with an insulating PTFE layer. The goldleaf electrode (GLE) was patterned using laser ablation and characterized by cyclic voltammetry, electrochemical impedance spectroscopy, scanning electronic microscopy, contact angle and 3D profiling. The GLEs were modified by a self-assembled mercaptopropionic monolayer, followed by surface activation to allow binding of the specific anti-E. coli antibody via carbodiimide linking. The biosensor showed a detection limit of 2 CFU/mL and a linear dynamic range of 10-107 CFU/mL for E. coli cells. No false positive signals were obtained from control bacteria. The obtained results demonstrated suitability of GLE for use in biosensors with high reliability and reproducibility. It is foreseeable that our work will inspire design of point-of-need biosensors broadly applicable in low-resource settings.

Experimental Methods for the Biological Evaluation of Nanoparticle-Based Drug Delivery Risks.

Many novel medical therapies use nanoparticle-based drug delivery systems, including nanomaterials through drug delivery systems, diagnostics, or physiologically active medicinal products. The approval of nanoparticles with advanced therapeutic and diagnostic potentials for applications in medication and immunization depends strongly on their synthesizing procedure, efficiency of functionalization, and biological safety and biocompatibility. Nanoparticle biodistribution, absorption, bioavailability, passage across biological barriers, and biodistribution are frequently assessed using bespoke and biological models. These methods largely rely on in vitro cell-based evaluations that cannot predict the complexity involved in preclinical and clinical studies. Therefore, assessing the nanoparticle risk has to involve pharmacokinetics, organ toxicity, and drug interactions manifested at multiple cellular levels. At the same time, there is a need for novel approaches to examine nanoparticle safety risks due to increased constraints on animal exploitation and the demand for high-throughput testing. We focus here on biological evaluation methodologies that provide access to nanoparticle interactions with the organism (positive or negative via toxicity). This work aimed to provide a perception regarding the risks associated with the utilization of nanoparticle-based formulations with a particular focus on assays applied to assess the cytotoxicity of nanomaterials.

Recent Advances on Peptide-Based Biosensors and Electronic Noses for Foodborne Pathogen Detection.

Foodborne pathogens present a serious issue around the world due to the remarkably high number of illnesses they cause every year. In an effort to narrow the gap between monitoring needs and currently implemented classical detection methodologies, the last decades have seen an increased development of highly accurate and reliable biosensors. Peptides as recognition biomolecules have been explored to develop biosensors that combine simple sample preparation and enhanced detection of bacterial pathogens in food. This review first focuses on the selection strategies for the design and screening of sensitive peptide bioreceptors, such as the isolation of natural antimicrobial peptides (AMPs) from living organisms, the screening of peptides by phage display and the use of in silico tools. Subsequently, an overview on the state-of-the-art techniques in the development of peptide-based biosensors for foodborne pathogen detection based on various transduction systems was given. Additionally, limitations in classical detection strategies have led to the development of innovative approaches for food monitoring, such as electronic noses, as promising alternatives. The use of peptide receptors in electronic noses is a growing field and the recent advances of such systems for foodborne pathogen detection are presented. All these biosensors and electronic noses are promising alternatives for the pathogen detection with high sensitivity, low cost and rapid response, and some of them are potential portable devices for on-site analyses.

Comparative meta-analysis of antimicrobial resistance from different food sources along with one health approach in Italy and Thailand.

Antimicrobial resistance (AMR) is increasing worldwide due to overuse, misuse and incomplete treatment of antibiotics. Many countries are facing the excessive issue due to the spreading of AMR not only in humans and animals, but also in water and agri-food sector. Our main aim was to perform a competitive meta-analysis of surveillance-resistant microbes and their antimicrobial superintendence in Italy and Thailand. Data have been collected from reports published for the period 2012-2021. A total of 9507 and 11,753 food samples contained 3905 (41.07%) and 3526 (30%) AMR bacteria in Italy and Thailand, respectively. In Italy, the highest microbial prevalence was ß-lactam and tetracycline, while in Thailand mostly isolates showed resistance to cephalosporin and aminoglycoside. Our findings contribute to highlighting the increment of AMR related to different microbes with tendency to become multidrug resistant.

Exploring Disease Management and Control through Pathogen Diagnostics and One Health Initiative: A Concise Review.

The "One Health" initiative is a critical strategy that recognizes the interconnectedness between human, animal, and environmental health in the spread and containment of infectious pathogens. With the ease of global transportation, transboundary disease outbreaks pose a significant threat to food safety and security, endangering public health and having a negative economic impact. Traditional diagnostic techniques based on genotypic and phenotypic analyses are expensive, time-consuming, and cannot be translated into point-of-care tools, hindering effective disease management and control. However, with advancements in molecular methods, biosensors, and new generation sequencing, rapid and reliable diagnostics are now available. This review provides a comprehensive insight into emergent viral and bacterial pathogens and antimicrobial resistance, highlighting the importance of "One Health" in connecting detection and effective treatment. By emphasizing the symbiotic relationship between human and animal health, this paper underscores the critical role of "One Health" initiatives in preventing and controlling infectious diseases.

Naked-eye detection of Staphylococcus aureus in powdered milk and infant formula using gold nanoparticles.

Nonspecific binding of proteins from complex food matrices is a significant challenge associated with a biosensor using gold nanoparticles (AuNPs). To overcome this, we developed an efficient EDTA chelating treatment to denature milk proteins and prevent their adsorption on AuNPs. The use of EDTA to solubilize proteins enabled a sensitive label-free apta-sensor platform for colorimetric detection of Staphylococcus aureus in milk and infant formula. In the assay, S. aureus depleted aptamers from the test solution, and the reduction of aptamers enabled aggregation of AuNPs upon salt addition, a process characterized by a color change from red to purple. Under optimized conditions, S. aureus could be visually detected within 30 min with the detection limit of 7.5 × 104 CFU/mL and 8.4 × 104 CFU/mL in milk and infant formula, respectively. The EDTA treatment provides new opportunities for monitoring milk contamination and may prove valuable for biosensor point-of-need applications.

Antioxidant and cell-friendly Fe2TiO5 nanoparticles for food packaging application.

An emerging technology of active packaging enables prolongation of food shelf life by limiting the oxygen transfer and the reactivity of free radicals, which both destruct food freshness. In this work, Fe2TiO5 nanoparticles were synthesized using a modified sol-gel method and evaluated as an enforcement of alginate food packaging film. Pure phase Fe2TiO5 nanoparticles had an average particle size of 44 nm and rhombohedral morphology. Fe2TiO5 nanoparticles induce no cell damage of human Caco-2 epithelial cells and show no inhibitory effect towards growth of a panel of bacterial strains, suggesting good biocompatibility. Films obtained by incorporation of Fe2TiO5 nanoparticles into alginate using the solvent casting method show no migration of iron or titanium ions from films to food simulants again suggesting their safety as a packaging material. Fe2TiO5 nanoparticles also showed strong antioxidant efficiency as determined using the DPPH assay, and confirmed further in a preservation test on fresh fruit.

Specific and sensitive detection of Influenza A virus using a biotin-coated nanoparticle enhanced immunomagnetic assay.

This study reports the development of a sensitive magnetic bead-based enzyme-linked immunoassay (MELISA) for the pan-reactive detection of the Influenza A virus. The assay combines immunomagnetic beads and biotin-nanoparticle-based detection to quantify a highly conserved viral nucleoprotein in virus lysates. At the capture step, monoclonal antibody-coated magnetic microbeads were used to bind and concentrate the nucleoprotein in samples. The colorimetric detection signal was amplified using biotinylated silica nanoparticles (NP). These nanoparticles were functionalized on the surface with short DNA spacers bearing biotin groups by an automated supported synthesis method performed on nano-on-micro assemblies with a DNA/RNA synthesizer. A biotin-nanoparticle and immunomagnetic bead-based assay was developed. We succeeded in detecting Influenza A viruses directly in the lysis buffer supplemented with 10% saliva to simulate the clinical context. The biotin-nanoparticle amplification step enabled detection limits as low as 3 × 103 PFU mL-1 and 4 × 104 PFU mL-1 to be achieved for the H1N1 and H3N2 strains respectively. In contrast, a classical ELISA test based on the same antibody sandwich showed detection limit of 1.2 × 107 PFU mL-1 for H1N1. The new enhanced MELISA proved to be specific, as no cross-reactivity was found with a porcine respiratory virus (PRRSV). Graphical abstract.

Applications of biosensors for bacteria and virus detection in food and water-A systematic review.

Biosensors for sensitive and specific detection of foodborne and waterborne pathogens are particularly valued for their portability, usability, relatively low cost, and real-time or near real-time response. Their application is widespread in several domains, including environmental monitoring. The main limitation of currently developed biosensors is a lack of sensitivity and specificity in complex matrices. Due to increased interest in biosensor development, we conducted a systematic review, complying with the PRISMA guidelines, covering the period from January 2010 to December 2019. The review is focused on biosensor applications in the identification of foodborne and waterborne microorganisms based on research articles identified in the Pubmed, ScienceDirect, and Scopus search engines. Efforts are still in progress to overcome detection limitations and to provide a rapid detection system which will safeguard water and food quality. The use of biosensors is an essential tool with applicability in the evaluation and monitoring of the environment and food, with great impact in public health.

Rapid point-of-need detection of bacteria and their toxins in food using gold nanoparticles.

Biosensors need to meet the rising food industry demand for sensitive, selective, safe, and fast food safety quality control. Disposable colorimetric sensors based on gold nanoparticles (AuNPs) and localized surface plasmon resonance are low-cost and easy-to-perform devices intended for rapid point-of-need measurements. Recent studies demonstrate various facile and versatile AuNPs-based analytical platforms for the detection of bacteria and their toxins in milk, meat, and other foods. In this review, we introduce the general characteristics and mechanisms of AuNPs calorimetric biosensors, and highlight optimizations needed to strengthen and improve the quality of devices for their application in food matrices.

Enrichment Free qPCR for Rapid Identification and Quantification of Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis in Chicken Meat Samples by a New Couple of Primers.

Campylobacter is the main cause of bacterial foodborne disease and poultry meat is the principal source of human infections. Rapid methods for Campylobacter detection are urgently needed to decrease high bacterial prevalence in poultry products. In this study, we developed new primers, CampyPFw and CampyPRv, that target the 16S-23S rRNA genes of Campylobacter jejuni, C. coli, C. lari and C. upsaliensis. The primers were tested on positive and negative reference strains in pure cultures and in inoculated poultry meat samples before their application in real-time PCR (qPCR) protocol for analyzing chicken meat samples. In parallel, the samples were tested by using the ISO 10272-1:2006 method. The qPCR protocol based on CampyPFw and CampyPRv showed good sensitivity, with the limit of detection of 4.6 × 102 cells/mL in chicken samples without enrichment steps.