In this study, we developed a lateral flow immunoassay (LFIA) for detecting duck adulteration in meat products. The assay employs a sandwich format targeting the molecular biomarker duck immunoglobulin (IgY) for accurate species identification. The system demonstrates high specificity and sensitivity, with no observed cross-reactivity among six commonly encountered mammalian and avian species. It is capable of detecting duck meat adulteration at levels as low as 1

BACKGROUND:Antibiotic fermentation residues, which contain antibiotic remnants and other potentially hazardous components, present increasing risks when illegally incorporated into animal feed. Their addition can promote the spread of antimicrobial resistance, drug accumulation, and ultimately threaten food safety and public health. Although regulatory control is required, existing analytical methods suffer from poor specificity, low accuracy, and limited applicability in practical settings. Therefore, there remains an urgent need for a rapid, accurate, and practical detection method to identify antibiotic fermentation residues directly within diverse feed matrices. RESULTS:We established three real-time PCR detection systems by designing primer-probe sets targeting OxyA, NeoN and AveD, the core biosynthetic genes of oxytetracycline-, neomycin- and avermectin-producing strains. The assay workflow is streamlined and requires only simple DNA extraction, without the need for chemical cleanup or feed matrix pretreatment. All reactions were completed within 2 h, demonstrating high operational efficiency suitable for routine monitoring. Sensitivity evaluation showed that fermentation residues could be reliably detected at a minimum level of 1% (w/w) in feed, and no cross-amplification occurred with non-target antibiotic residues or diverse feed ingredients. Artificially adulterated samples verified robust applicability across matrices including soybean meal, cottonseed meal and compound feed additives. Repeatability assessment further confirmed excellent stability, with intra- and inter-assay CV values maintained below 2%. These results collectively demonstrate that the developed assays are rapid, accurate and highly adaptable to real feed-testing environments. SIGNIFICANCE:This work represents the first demonstration of real-time PCR detection of multiple antibiotic fermentation residues in feed based on strain-origin specific biosynthetic genes. The method provides a rapid, sensitive, accurate, and high-specificity molecular tool for on-site regulatory screening, enabling early identification of the illicit addition of antibiotic fermentation residues. The application of this technology strengthens feed safety surveillance and contributes to preventing antibiotic-related hazards and safeguarding public health.

Bioactive packaging made from biopolymers can decrease pollution in the environment in addition to prolonging the shelf life of chilled beef. A novel composite antibacterial preservative film was prepared using gelatin (Gel)/chitosan (Ch) as the film material and Litsea cubeba essential oil (LCEO) as the antibacterial agent. Its physical properties, structure, antioxidant activity, and antibacterial performance were studied. When the ratio of gelatin to chitosan was 6:4, the film exhibited good tensile strength, flexibility, and water resistance. Adding 2% LCEO to the film material further enhanced its tensile strength and improved the composite film's hydrophobicity, antioxidant activity, and antibacterial performance. The water solubility and moisture content of the composite film were reduced to 29.73% and 14.91%, respectively, while the antioxidant activity increased to 82.86%. The composite film with 2% LCEO was applied to preserve chilled beef. Compared to the composite film without essential oils, the 10th day pH, thiobarbituric acid reactive substances (TBARS), and total viable count (TVC) decreased by 0.48, 0.34 mg MDA/kg, and 1.83 log CFU/g, respectively. This indicates that the composite film containing Litsea cubeba essential oil has an excellent preservative effect on beef, extending the shelf life of chilled beef by 6 days.

Accurately detecting viable foodborne pathogenic bacteria is essential for food safety risk assessments and public health interventions. Traditional plate counting is time-consuming and operationally cumbersome. Immunological assays are unable to distinguish viable from dead cells, whereas conventional nucleic acid amplification is often affected by residual DNA originating from dead bacteria. These limitations render conventional approaches inadequate for rapid and precise field detection. Functional nucleic acids (FNAs) offer a promising alternative for viability detection because of their high sensitivity, specificity, target diversity, and programmable integrability. This review provides a systematic overview of molecular recognition strategies and FNA-based detection technologies for identifying viable foodborne microorganisms. We categorize the biomarkers targeted by FNAs into nucleic acids, surface structures, and metabolic activities. Building on this categorization, we examine the core principles and technological evolution of primers, aptamers, DNAzymes, guide nucleic acids, and oligonucleotide probes in viability discrimination. We then outline the practical applications of these technologies across the food supply chain and discuss the remaining challenges and future directions in the field. Ultimately, this work provides a theoretical reference and practical guidance for ensuring food safety and advancing precise microbial risk management.

Acetylcholinesterase inhibition-luciferase bioluminescence system enables rapid detection of organophosphate and carbamate pesticide residues. However, firefly luciferase (FLuc) suffers from poor stability and short luminescence duration, thereby introducing uncertainties in detection. To address this problem, nanoflower-immobilized FLuc (FLuc@NFs) was prepared using a coprecipitation method, which markedly enhanced stability and activity. FLuc@NFs exhibited intact nanoflower structures and allowed storage at 25 °C and at 4 °C for 2 weeks, overcoming the limitation of free enzyme storage at -20 °C. In the bioluminescence reaction, FLuc@NFs at 1 mg/mL achieved 62.14 % higher luminescence than free FLuc, with stable signals extending from 10 s to 30 min, enabling high-throughput detection. The system achieved detection limits of 5 and 10 ng/mL for chlorpyrifos and carbaryl, respectively, demonstrating improved reliability and sensitivity in pesticide detection.

Simultaneous determination of Somatic Cell Count (SCC) and Total Plate Count (TPC) in raw milk reflects milk cleanliness and cow health. The luciferase was thermally unstable and required storage at -20 °C, with repeated freeze-thaw cycles reducing activity. To overcome this limitation, an inorganic hybrid nanoflower luciferase was synthesized by mixing genetically engineered free luciferase with PBS and zinc acetate, followed by incubation, centrifugation, washing, and resuspension. Transmission electron microscopy confirmed successful assembly of luciferase into nanoflower structures. The nanoflower luciferase remained stable over 75 % of its initial activity after 21 days at 4 °C, whereas free enzyme required -20 °C.Compared with genetically engineered free luciferase, the nanoflower form effectively and simultaneously detected SCC and TPC in raw milk even after 2-fold dilution. The limits of quantification were 1 × 104 cells/mL for SCC and 1 × 105 CFU/mL for TPC.

Probiotics are beneficial microorganisms that support host health. They are extensively used in food, healthcare and pharmaceutical industries. Accurate viability assessment is imperative not only to ensure product quality and efficacy but also to meet consumer expectations. This review provides a systematic analysis of probiotic history, international regulatory standards and functional characteristics across strains. It provides a comprehensive overview of probiotic enumeration techniques, which range from traditional culture methods to modern emerging technologies that drive current research and applications. By critically evaluating the strengths and limitations of these methods, this review explores future directions for probiotic enumeration, offering valuable insights for quality control in probiotic products and supporting advancements in related scientific research.

BACKGROUND:Chilled beef is valued for its nutrition and tenderness but is vulnerable to contamination and spoilage during refrigeration. This study applied non-targeted metabolomics (ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS)) to track metabolite changes and their impact on beef quality across storage days 0, 3, 5, 7, and 10. RESULTS:A total of 63 potential marker compounds were identified, with amino acid, carbohydrate, and nucleotide degradation as the main drivers of spoilage and flavor change. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment revealed purine, pentose, and amino acid metabolism as key pathways. Pearson correlation analysis highlighted hypoxanthine, citric acid, xanthosine, tyrosine, phenylalanine, xanthine, d-glucose-6-phosphate, and inosine 5'-monophosphate as strongly associated with quality decline, suggesting their potential as biomarkers of spoilage. CONCLUSION:Metabolomics effectively captured biochemical dynamics during beef refrigeration. Identified pathways and metabolites provide valuable insights into spoilage mechanisms, with several compounds serving as promising biomarkers for monitoring quality changes. © 2025 Society of Chemical Industry.

As a slaughter by-product, animal keratin exhibits a wide range of bioactive functions, including antioxidant, immunomodulatory, and skin-repair-promoting effects, making it an important biomaterial with excellent biocompatibility and significant application potential. Bioactive peptides derived from animal keratin can enhance physiological functions and improve health through multiple mechanisms, and these bioactivities have attracted increasing research attention in the fields of food, medicine, and cosmetics in recent years. While considerable research has focused on the physicochemical properties of keratin, substantial challenges remain in its functional development, extraction and processing, and commercialization. This review provides an in-depth analysis of the nutritional functions, extraction and processing techniques, and current status of product development for animal keratin, and discusses its potential applications and future research directions, offering a reference for its further development.

Feed safety is critical to the quality of livestock and poultry products and plays a key role in ensuring food safety and public health. To address the growing issue of illicit addition of antibiotic fermentation residues, such as penicillin mycelial waste, into feed, we developed a real-time PCR method targeting the orf70c gene, a specific marker within the biosynthetic gene cluster of industrial penicillin-producing strains. Specific primers and a probe were designed to enable accurate detection across nine types of feed. The assay requires no complex sample pretreatment and completes the entire process from DNA extraction to result within 2 h. It reliably detects as little as 0.5 % (w/w) of penicillin fermentation residue in feed. The method demonstrated high reproducibility, with inter-assay variation between 0.46 similar to 0.55 % and intra-assay variation between 0.37 similar to 0.69 %. To our knowledge, this is the first real-time PCR assay developed for detecting antibiotic fermentation residues in feed. The method provides a practical tool for feed safety monitoring and supports sustainable practices in the feed and animal production industries.

Edible oil adulteration is a significant food safety concern in today's food fraud landscape. DNA-based detection methods emerging as the most reliable approach for identifying adulterated products. However, the processed oil matrix contains only trace amounts of highly degraded DNA, which significantly increases the difficulty of DNA extraction and greatly reduces the success rate of PCR amplification. To address these technical challenges, we designed species-specific primers for amplicons of varying lengths and systematically evaluated their performance in real-time PCR using DNA extracted from soybean, peanut, and rapeseed oils, followed by application in adulterated oil detection. All primer sets demonstrated high specificity, sensitivity and, most importantly, equal amplification efficiency. Results indicated that shorter amplicons were consistently detectable across all three vegetable oils, while longer amplicons were more likely to yield false negatives in real-time PCR reactions. Soybean oil showed CT values of 29 (74 bp) versus 38 (170 bp), peanut oil exhibited a CT value of 31 (54 bp) with no amplification for 86 bp, and rapeseed oil displayed a CT value of 34 (66 bp) with no signal for 101 bp. The use of shorter primers successfully facilitated the detection of adulterated oil.These findings suggest that amplicon length being a critical determinant for reliable DNA-based identification and authentication of edible oils. Thus, designing amplicons within the 50-80 bp range can significantly enhance the success rate of DNAbased identification and authentication of edible oils.

Organophosphorus pesticides (OPs) and carbamate pesticides (CBs) are the most commonly used pesticides in agricultural cultivation. The pesticide residues in plant products can easily enter dairy cows through feed, resulting in the pesticide low-concentration residues in milk. Traditional acetylcholinesterase (AChE) inhibitionbased colorimetric methods have low sensitivity and could not satisfy the detection of low-concentrations of OPs and CBs residues in dairy products. In this study, we combined firefly luciferase (FLuc)-mediated bioluminescence with the inhibition of AChE-catalyzed substrate activity by pesticides to develop a highly sensitive and rapid method for detecting OPs and CBs residues in milk. AChE breaks down D-luciferin acetate to produce Dluciferin, which is recognized by the FLuc and emits luminescence in the presence of ATP. However, the presence of OPs and CBs inhibits AChE, causing the reduction or disappearance of the luminescence signal. The luminescence signal can be detected using a hand-held luminescence photometer, eliminating the need for large instrumentation. The AChE-FLuc system accurately detected OPs and CBs in milk within 30 min, with detection limits of 7.89 ng/mL and 1.75 ng/mL, respectively. The sensitivity of this method is approximately ten times higher than that of traditional AChE inhibition methods, meeting the pesticide residue limits in milk set by China and the European Union.

Probiotics play a critical role in maintaining or improving gut microbiome balance, offering numerous health benefits. Therefore, the accurate quantification of viable probiotics is essential for ensuring the efficacy of probiotic products. However, traditional culture‐based methods are cumbersome and time‐intensive, limiting their applications in scenarios requiring real‐time decisions. Here, we report a portable, cost‐effective system (approximately $2 per test, including reagents and one‐time microfluidic chip usage) that enables rapid (3 min) on‐site quantification of viable probiotics through a microfluidic chip integrated with a fluorescent biosensor and advanced image recognition technology. The system's reliability and accuracy were validated by comparison with the standard flow cytometry and plate counting, with results showing no significant differences in viable probiotics numbers. Furthermore, the platform demonstrated excellent specificity in distinguishing dead probiotics from viable ones with a large quantification range of 10 7 –10 11 colony‐forming units (CFU) mL −1 , which encompasses the concentration range of commonly encountered probiotics, thereby meeting practical detection requirements. In summary, this portable system offers an innovative solution for rapid, on‐site identification, and precise quantification of live and dead probiotics. Given these attributes, the system demonstrates significant commercial potential, particularly in situations requiring immediate results and efficient decision‐making.

Cattle-yaks are increasingly cultivated to boost yak dairy production while reports claim that will decrease the nutritional value of yak milk. However, studies on protein composition differences, especially functional proteins, between yak and cattle-yak milk are limited. This study explores these differences using data-independent acquisition (DIA) proteomics to compare whey proteins and milk fat globule membrane (MFGM) proteins in both types of milk. The results showed most up-regulated differentially expressed proteins (DEPs) in cattle-yak milk are more involved in promoting calcium absorption, immune regulation, and intestinal health which are essential nutrients for the growth and development of infants. Notably, the most important functional protein lactotransferrin and osteopontin content in cattle-yak milk are 8.03 and 3.44 times higher than that in yak milk respectively. This study sheds light on the potential crossbreeding advantages of cattle-yak milk and provides scientific evidence for cattle-yak breeding and the exploration of its milk.

Pingliang Red Cattle, a renowned geographical indication product in China, is distinguished by its superior meat quality, yet the scientific basis for its unique attributes remains underexplored. This study integrated metabolomic and transcriptomic analyses to elucidate the biochemical and physiological factors underlying the enhanced flavor, color stability, and tenderness of Pingliang Red Cattle beef compared to Qinchuan and Simmental cattle. Metabolomic profiling revealed significantly elevated levels of inosine monophosphate (IMP, 2.86–3.96× higher) and glutathione (GSH, 2.42–5.43× higher) in Pingliang Red Cattle, contributing to intense umami flavor and prolonged meat color retention. Notably, ergothioneine (EGT), a potent antioxidant, was identified for the first time in Pingliang Red Cattle beef, with concentrations 2.55× and 4.25× higher than in Qinchuan and Simmental, respectively. Transcriptomic analysis highlighted the upregulation of 21 tenderness-related genes (e.g., FABP3, PRDX6, CAST) and key enzymes in purine and glutathione metabolism pathways (e.g., PDE4D, ADSL, GGT1), correlating with meat tenderness and the improved meat quality. Additionally, Pingliang Red Cattle’s natural forage-rich diet and low-density rearing practices were critical in enhancing these traits. These findings provide a scientific foundation for Pingliang Red Cattle’s premium quality, offering actionable insights for GI product branding, quality optimization, and market competitiveness. The multi-omics approach established here serves as a paradigm for quality assessment and improvement of other GI agricultural products, bridging traditional reputation with molecular evidence.

Adulteration of meat is a global issue, necessitating rapid, inexpensive, and simple on-site testing methods. Therefore, the present study aimed to develop a one-minute toothpick-based DNA extraction method, a handheld microfluidic chip, and a smartphone-controlled portable analyzer for detecting multiple meat adulterations. A toothpick was inserted into the meat to promote DNA release and adsorption. Furthermore, a handheld micro- fluidic chip was designed for DNA elution on toothpicks and fluid distribution. Finally, a smartphone-actuated portable analyzer was developed to function as a heater, signal detector, and result reader. The portable device comprises a microcontroller, a fluorescence detection module, a step scanning unit, and a heating module. The proposed device is portable, and the app is user-friendly. This simple design, easy operation, and fast- response system could rapidly detect as little as 1% of simulated adulterated samples (following UK standards) within 40 min at a cost of less than USD 1 per test.

With the globalization and complexity of the food supply chain, the market is becoming increasingly competitive and food fraudulent activities are intensifying. The current state of food detection faced two primary challenges. Firstly, existing testing methods were predominantly laboratory-based, requiring complex procedures and precision instruments. Secondly, there was a lack of accurate and efficient quantitative detection methods. Taking cow's milk as an example, this study introduced a novel method for nucleic acid quantification in dairy products, based on lateral flow strips (LFS). The core idea of this method is to design single-stranded DNA (ssDNA) probes to hybridize with mitochondrial genes, which are abundant, stable, and species-specific in dairy products, as detection targets. Drawing inspiration from the principles of nucleic acid amplification, this research innovatively established a new DNA hybridization method, named LAMP-Like Hybridization (HybLAMP-Like). Leveraging the denaturation and DNA polymerization functions of the bst enzyme, efficient binding of the probe and template strand was achieved. This method eliminated the need for nucleic acid amplification, simplifying the procedure and mitigating aerosol contamination, thereby ensuring the accuracy of the detection results. The method exhibited exceptional sensitivity, capable of detecting extremely low to 12.5 ng in visual inspection and 3.125 ng when using a reader. In terms of practicality, it could achieve visual detection of cow's milk content as low as 1% in adulterated dairy products. When combined with a portable LFS reader, it also enabled precise quantitative analysis of milk adulteration.

Species identification has become a significant concern due to the growing use of food alternatives that may cause allergies and reduce nutritional value. To address the issue of fraudulent adulteration of goat milk products with cow milk, we have developed an affordable, portable, and user-friendly platform called microfluidicintegrated nucleic acid lateral flow strips (LFS). This platform enables simultaneous detection of components derived from both goats and cows in goat milk. In this study, we have introduced an innovative nucleic acid labeling method. The loop primers of loop-mediated isothermal amplification (LAMP) have been modified with amplification terminator spacer C3 and an oligonucleotide sequence, thus eliminating the requirement for costly antibodies in traditional nucleic acid LFS. This modification not only lowers costs but also enables multiple detections. Additionally, we have integrated the LAMP and LFS assay steps into a microfluidic chip, allowing convenient on-site detection while effectively preventing aerosol contamination of LAMP products. The testing process includes rapid DNA extraction, followed by a short nucleic acid addition and incubation for visualized results in about 50 min. This platform is user-friendly, requiring no specialized equipment or extensive training, making it suitable for rapid on-site detection of dairy products by personnel in diverse fields.

Genetically modified food has come under suspicion because of the potential safety risks. Only a limited number of target genes could be detected by traditional methods. Here, customized-fast DNA extraction methods and a LAMP (Loop-mediated isothermal amplification) system were developed combining a hand-held chip, portable analyzer, and an Android app for the qualitative detection of genetically modified crops. DNA of seeds and leaves was obtained within 5minutes. Primers were designed with high specificity and sensitivity (10 copies) for later LAMP amplification. Crude DNA was added to the chip pre-embedded with customizable primers by quantitative dropper and the whole detection process was less than 40minutes without any large-scale instrument. The limit of detection was approximately 0.1% (w/w). The detection system with its low workload, high accuracy, and stability offers a novel, effective approach for the rapid detection of GM corps and many other food safety areas in the field.

Monitoring and evaluating food quality, especially meat quality, has received a growing interest to ensure human health and decrease waste of raw materials. Standard analytical approaches used for meat spoilage assessment suffer from time consumption, being labor-intensive, operation complexity, and destructiveness. To overcome shortfalls of these traditional methods and monitor spoilage microorganisms or related metabolites of meat products across the supply chain, emerging analysis devices/systems with higher sensitivity, better portability, on-line/in-line, non-destructive and cost-effective property are urgently needed. Herein, we first overview the basic concepts, causes, and critical monitoring indicators associated with meat spoilage. Then, the conventional detection methods for meat spoilage are outlined objectively in their strengths and weaknesses. In addition, we place the focus on the recent research advances of emerging non-destructive devices and systems for assessing meat spoilage. These novel strategies demonstrate their powerful potential in the real-time evaluation of meat spoilage.