Effect of lignin on the formation of polycyclic aromatic hydrocarbons in smoked and grilled meat products.

In order to explore the influence of wood types on formation of polycyclic aromatic hydrocarbons (PAHs) in traditional smoked and grilled meat products, the effect of lignin in woods on formation of PAHs was investigated in meat model systems. The results showed that PAHs formation was much dependent on the heating conditions. The addition of lignin led to significantly increased PAHs, which being connected with lignin structure. In comparison, the formation of PAHs was more facilitated by lignin with G structure than that with G/S structure. However, further study of adding lignin precursors demonstrated that lignin precursors with S structure were more favorable to the formation of PAHs than those with G structure. It was proposed that the relative content and activity of G/S structure of lignin in wood played a significant role in the formation of PAHs, which might provide theoretical reference for inhibition of PAHs fundamentally.

Development of fluorescent GO-AgNPs-Eu3+ nanoparticles based paper visual sensor for foodborne spores detection.

Foodborne spores are ubiquitous with extremely strong resistance, and pose a serious threat to food safety and human health. Therefore, rapid, sensitive, and selective detection of spores are crucial. In this study, a fluorescent probe was developed based on lanthanide ion (Eu3+)-labeled nano-silver-modified graphene oxide (GO-AgNPs-Eu3+) for the detection of 2,6-dipicolinic acid (DPA), a biomarker unique to spores, to allow quantitative spores detection. The GO-AgNPs-Eu3+ nano-fluorescent probe was loaded onto a polyvinylidene fluoride microfiltration membrane, and a smartphone-assisted portable GO-AgNPs-Eu3+ nanoparticles-based paper visual sensor was designed for rapid on-site quantitative and real-time online detection of spores. The results indicated that the developed probe achieved equilibrium binding with DPA within 5 min, and enhanced fluorescence emission through antenna effect. The fluorescence detection presented a good linear relationship in the DPA concentration range of 0-45 µM, with a DPA detection limit of 4.62 nM and spore detection limit of 104 cfu/mL. The developed sensor showed a change in fluorescence from blue to red with increasing DPA concentration, and this color change was quantitatively detected through smartphone RGB variations, with a detection limit of 13.1 µM for DPA and 6.3 cfu/mL for Bacillus subtilis spores. Subsequently, the sensitivity and selectivity of the developed sensor were verified using actual milk and water samples spiked with B. subtilis spores. The results of this study provided objective technological support for rapid detection of spores, which is important for reducing the occurrence of foodborne diseases and improving food safety.

Selective purification and rapid quantitative detection of spores using a "stepped" magnetic flow device.

A study on the inactivation and germination mechanism of spores is very important in the application of spores, as such high-purity spores are the basis of related research. However, spores and vegetative cells of bacteria often coexist, and it is difficult to separate them. In this study, a magnetic flow device for the purification of spores in the culture medium system was developed based on a "stepped" structure with a magnetic force that could absorb vegetative cells with magnetic nanoparticles. The operation process was as follows: first, vancomycin functionalized nanoparticles were used to prepare Van-Fe3O4 NPs, which were then combined with vegetative cells to form a magnetic conjugate. Subsequently, the magnetic conjugate (vegetative cells) flowed through the "stepped" magnetic flow device and was adsorbed. Meanwhile, the spores moved through the channel and were collected. The achieved purity of the collected spores was more than 95%. Further, the number of the obtained spores was quickly quantified using Raman spectroscopy. The entire purification and quantitative process can be completed within 30 min and the limit of detection was 5 CFU mL-1. This study showed outstanding spore purification ability and provided a new method for purification and rapid quantitative detection of spores.

Effect of Molecular Weight on the Structural and Emulsifying Characteristics of Bovine Bone Protein Hydrolysate.

The emulsifying capacity of bovine bone protein extracted using high-pressure hot water (HBBP) has been determined to be good. Nevertheless, given that HBBP is a blend of peptides with a broad range of molecular weights, the distinction in emulsifying capacity between polypeptide components with high and low molecular weights is unclear. Therefore, in this study, HBBP was separated into three molecular weight components of 10-30 kDa (HBBP 1), 5-10 kDa (HBBP 2), and <5 kDa (HBBP 3) via ultrafiltration, and the differences in their structures and emulsifying properties were investigated. The polypeptide with the highest molecular weight displayed the lowest endogenous fluorescence intensity, the least solubility in an aqueous solution, and the highest surface hydrophobicity index. Analysis using laser confocal Raman spectroscopy showed that with an increase in polypeptide molecular weight, the α-helix and ß-sheet contents in the secondary structure of the polypeptide molecule increased significantly. Particle size, rheological characteristics, and laser confocal microscopy were used to characterize the emulsion made from peptides of various molecular weights. High-molecular-weight peptides were able to provide a more robust spatial repulsion and thicker interfacial coating in the emulsion, which would make the emulsion more stable. The above results showed that the high-molecular-weight polypeptide in HBBP effectively improved the emulsion stability when forming an emulsion. This study increased the rate at which bovine bone was utilized and provided a theoretical foundation for the use of bovine bone protein as an emulsifier in the food sector.

Construction of Graphene@Ag-MLF composite structure SERS platform and its differentiating performance for different foodborne bacterial spores.

A new surface-enhanced Raman spectroscopy (SERS) biosensor of Graphene@Ag-MLF composite structure has been fabricated by loading AgNPs on graphene films. The response of the biosensor is  based on plasmonic sensing. The results showed that the enhancement factor of three different spores reached 107 based on the Graphene@Ag-MLF substrate. In addition, the SERS performance was stable, with good reproducibility (RSD<3%). Multivariate statistical analysis and chemometrics were used to distinguish different spores. The accumulated variance contribution rate was up to 96.35% for the top three PCs, while HCA results revealed that the spectra were differentiated completely. Based on optimal principal components, chemometrics of KNN and LS-SVM were applied to construct a model for rapid qualitative identification of different spores, of which the prediction set and training set of LS-SVM achieved 100%. Finally, based on the Graphene@Ag-MLF substrate, the LOD of three different spores was lower than 102 CFU/mL. Hence, this novel Graphene@Ag-MLF SERS substrate sensor was rapid, sensitive, and stable in detecting spores, providing strong technical support for the application of SERS technology in food safety.

Effects of Different Bacteriostats on the Dynamic Germination of Clostridium perfringens Spores.

Bacteriostats, as chemical substances that inhibit bacterial growth, are widely used in the sterilization process; however, their effects on spindle spores are unclear. In this study, the effects of bacteriostats, including nine commonly used food additives and four detergents, on the growth of Clostridium perfringens spores were investigated. The results showed that 0.07‱ ethylenediaminetetraacetate had a good inhibitory effect on C. perfringens spore growth, and the spore turbidity decreased by 4.8% after incubation for 60 min. Furthermore, 0.3‱ tea polyphenols, 0.8‱ D-isoascorbic acid, and 0.75‱ potassium sorbate promoted leakage of contents during spore germination. Among the four detergents, 5‱ glutaraldehyde solution presented the best inhibitory effect on the growth of C. perfringens spores, and the spore turbidity decreased by 5.6% after incubation for 60 min. Further analysis of the inactivation mechanism of spores by the bacteriostats was performed by comparing the leakage of UV-absorbing substances during germination. The results revealed that bacteriostats could not directly kill the spores, but could inactivate them by inhibiting germination or damaging the spore structure during germination, thus preventing the formation of bacterial vegetative bodies. These findings provide important information and reference for the mechanism underlying the effects of different bacteriostatic agents on spore growth.

Effects of sporulation conditions on the growth, germination, and resistance of Clostridium perfringens spores.

Clostridium perfringens can form metabolically dormant spores that can survive in meat preservation processes and cause food spoilage and human disease upon germination and outgrowth. The characteristics of spores in food products are closely related to the sporulation environment. To control or inactivate C. perfringens spores in food industry, the effects of sporulation conditions on the spores characteristics should be examined. This study aimed to investigate the effects of temperature (T), pH, and water activity (aw) on the growth, germination, and wet-heat resistance of C. perfringens C1 spores isolated from food product. The results showed that C. perfringens C1 spores produced at T = 37 °C, pH = 8, and aw = 0.997 had the highest sporulation rate and germination efficiency and lowest wet-heat resistance. A further increase in pH and sporulation temperature reduced the spore counts and germination efficiency, but enhanced spores' wet-heat resistance. By using air-drying method and Raman spectroscopy analysis, the water content, composition, and levels of calcium dipicolinic acid, proteins, and nucleic acids in spores produced under different sporulation conditions were determined. The results obtained revealed that sporulation conditions should be carefully considered during food production and processing, thus providing a novel insight into prevention and control of spores in food industry.

Preparation of AgNPs self-assembled solid-phase substrate via seed-mediated growth for rapid identification of different bacterial spores based on SERS.

In this study, we developed a novel and sensitive AgNPs self-assembled solid-phase (AgNPs-SASP) SERS substrate platform using seed-mediated and liquid interface self-assembly methods and applied the platform for the detection of three bacterial spores. Multivariate statistical analysis (HCA and LDA) were employed for identification and categorization of the obtained data. The results illustrated that AgNPs-SASP exhibited high reproducibility and Raman enhancement effect (1.43 × 104). The Raman shift bands of the three bacterial spores ranged from 400 to 1800 cm-1 demonstrating the difference in reigon and intensity. And the intensities of their Ca2+-DPA Raman bands are significantly different. HCA results revealed that the spectra for the three bacterial spores were statistically different while LDA completely differentiated the spectra for the three bacterial spores with 100 % sensitivity and specificity. Overall, the novel SERS platform based on AgNPs-SASP provides an effective tool for food safety risk control and detection.

Preparation of an AgNPs@Polydimethylsiloxane (PDMS) multi-hole filter membrane chip for the rapid identification of food-borne pathogens by surface-enhanced Raman spectroscopy.

The consumption of food infected with food-borne pathogens has become a global public health problem. Therefore, it is monitor food-borne infections to avoid health and financial consequences. The rapid detection and differentiation of bacteria for biomedical and food safety applications continues to be a significant challenge. Herein, we present a label-free surface-enhanced Raman scattering approach for separating harmful bacteria from food. The method relies on the ascorbic acid reduction method to synthesize silver nanoparticles (AgNPs) and a polydimethylsiloxane (PDMS) multi-hole filter membrane chip (AgNPs@PDMS multi-hole filter membrane chip). Surface-enhanced Raman spectroscopy (SERS) was used, followed by multivariate statistical analysis to differentiate five important food-borne pathogens, including Staphylococcus aureus, Salmonella typhimurium, Listeria monocytogenes, Clostridium difficiles and Clostridium perfringens. The results demonstrated that compared to normal Raman signals, the intensity of the SERS signal was greatly enhanced with an analytical enhancement factor of 5.2 × 103. The spectral ranges of 400-1800 cm-1 were analyzed using principal component analysis (PCA) and stepwise linear discriminant analysis (SWLDA) were used to determine the optimal parameters for the discrimination of food-borne pathogens. The first three principal components (PC1, PC2, and PC3) accounted for 87.3% of the total variance in the spectra. The established SWLDA model had 100% accuracy and cross-validation accuracy, which accurately distinguished the SERS spectra of the five species. In conclusion, the SERS technology based on the AgNPs@PDMS multi-hole filter membrane chip was useful for the rapid identification of food-borne pathogens and can be employed for food quality management.

Novel Insights into Total Flavonoids of Rhizoma Drynariae against Meat Quality Deterioration Caused by Dietary Aflatoxin B1 Exposure in Chickens.

Aflatoxin B1 (AFB1) is a group of highly toxic mycotoxins that are commonly found in human and animal foods and threaten animal and human food safety. Total flavonoids of Rhizoma Drynaria (TFRD), a traditional Chinese medicinal herb, exert multiple biological activities such as immunomodulatory, anti-inflammatory, and anti-oxidation effects. Here, a total of 160 healthy 21-day-old male broilers were randomly divided into four groups: the CON group, the TFRD group, the AFB1 group, and the AFB1 + TFRD group. The study found that AFB1 exposure altered the breast meat quality-related indicators, including meat sensory and physical indicators. Metabolomics analysis further showed that the change in meat quality was closely associated with significantly differential metabolites of breast muscle. Furthermore, spotlighted amino acid content contributes to changes in the secondary structure of the myofibrillar protein by Raman spectroscopy analysis, which was associated with the oxidative stress and inflammatory response in AFB1-exposed breast meat. Meanwhile, dietary 125 mg/kg TFRD supplementation could effectively restore the changes in breast meat quality. Taken together, these results by multi-technical analysis revealed that AFB1 exposure causes deterioration of chicken meat quality and that TFRD may be a potential herbal extract to antagonize mycotoxicity.

Near-infrared spectroscopy coupled with chemometrics algorithms for the quantitative determination of the germinability of Clostridium perfringens in four different matrices.

Clostridium perfringens (C. perfringens) has the ability to form metabolically-dormant spores that can survive food preservation processes and cause food spoilage and foodborne safety risks upon germination outgrowth. This study was conducted to investigate the effects of different AGFK concentrations (0, 50, 100, 200 mM/mL) on the spore germination of C. perfringens in four matrices, including Tris-HCl, FTG, milk, and chicken soup. C. perfringens spore germinability was investigated using near infrared spectroscopy (NIRS) combined with chemometrics. The spore germination rate (S), the OD600%, and the Ca2+-DPA% were measured using traditional spore germination methods. The results of spore germination assays showed that the optimum germination rate was obtained using 100 mM/L concentrations of AGFK in the FTG medium, and the S, OD600% and Ca2+-DPA% were 98.6%, 59.3% and 95%, respectively. The best prediction models for the S, OD600% and Ca2+-DPA% were obtained using SNV as the preprocessing method for the original spectra, with the competitive adaptive weighted resampling method (CARS) as the characteristic variables related to the selected spore germination methods from NIRS data. The results of the S showed that the optimum model was built by CARS-PLSR (RMSEV = 0.745, Rc = 0.897, RMSEP = 0.769, Rp = 0.883). For the OD600%, interval partial least squares regression (CARS-siPLS) was performed to optimize the models. The calibration yielded acceptable results (RMSEV = 0.218, Rc = 0.879, RMSEP = 0.257, Rp = 0.845). For the Ca2+-DPA%, the optimum model with CARS-siPLS yielded acceptable results (RMSEV = 44.7, Rc = 0.883, RMSEP = 50.2, Rp = 0.872). This indicated that quantitative determinations of the germinability of C. perfringens spores using NIR technology is feasible. A new method based on NIR was provided for rapid, automatic, and non-destructive determination of the germinability of C. perfringens spores.

Detection of triterpene acids distribution in loquat (Eriobotrya japonica) leaf using hyperspectral imaging.

Hyperspectral images (431-962nm) and partial least squares (PLS) were used to detect the distribution of triterpene acids within loquat (Eriobotrya japonica) leaves. 72 fresh loquat leaves in the young group, mature group and old group were collected for hyperspectral imaging; and triterpene acids content of the loquat leaves was analyzed using high performance liquid chromatography (HPLC). Then the spectral data of loquat leaf hyperspectral images and the triterpene acids content were employed to build calibration models. After spectra pre-processing and wavelength selection, an optimum calibration model (Rp=0.8473, RMSEP=2.61mg/g) for predicting triterpene acids was obtained by synergy interval partial least squares (siPLS). Finally, spectral data of each pixel in the loquat leaf hyperspectral image were extracted and substituted into the optimum calibration model to predict triterpene acids content of each pixel. Therefore, the distribution map of triterpene acids content was obtained. As shown in the distribution map, triterpene acids are accumulated mainly in the leaf mesophyll regions near the main veins, and triterpene acids concentration of young group is less than that of mature and old groups. This study showed that hyperspectral imaging is suitable to determine the distribution of active constituent content in medical herbs in a rapid and non-invasive manner.

A rapid and nondestructive method to determine the distribution map of protein, carbohydrate and sialic acid on Edible bird's nest by hyper-spectral imaging and chemometrics.

Edible bird's nest (EBN) is a precious functional food in Southeast Asia. A rapid and nondestructive method for determining the distribution map of protein content (PC), carbohydrate content (CC) and sialic acid content (SAC) on EBN sample was proposed. Firstly, 60 EBNs were used for hyperspectral image acquisition, and components content (PC, CC and SAC) were determined by chemical analytical methods. Secondly, the spectral signals of EBN hyperspectral image and EBN components content were used to build calibration models. Thirdly, spectra of each pixel in EBN hyperspectral image were extracted, and these spectra were substituted in the calibration models to predict the PC, CC and SAC of each pixel in the EBN image, so the visual distribution maps of PC, CC and SAC on the whole EBN were obtained. It is the first time to show the distribution tendency of PC, CC and SAC on the whole EBN sample.

Microfabricated interdigitated Au electrode for voltammetric determination of lead and cadmium in Chinese mitten crab (Eriocheir sinensis).

An in-situ plating bismuth modified interdigitated Au electrode (IAE) was developed for determination of Pb(2+) and Cd(2+) in crab. The IAE was fabricated and used as counter electrode and working electrode. Cyclic voltammetry (CV) was performed with the IAE for studying electrochemical performance, showing clear redox peaks. The key operational parameters were optimized, which were 600 µg L(-1) of Bi(3+) concentration, 0.1 mol L(-1) of acetate buffer (pH 4.5), -1.2V of deposition potential and 180 s of deposition time. Under optimized condition, the linear range of IAE was from 5 to 50 µg L(-1) for both metal ions, with detection limit (threefold signal-to-noise) of 0.74 µg L(-1) for Pb(2+) and 0.86 µg L(-1) for Cd(2+). Finally, the developed sensor was applied to detect Pb(2+) and Cd(2+) in crab extract solutions by standard addition method. The results were in good agreement with outcomes obtained by inductively coupled plasma mass spectrometry.