Effects of ohmic heating with different voltages on the quality and microbial diversity of cow milk during thermal treatment and subsequent cold storage.

Ohmic heating (OH), an innovative heating technology, presents potential applications in the pasteurization of liquid foods. Therefore, the study was conducted to evaluate the effect of OH at various voltage gradients (10 V/cm, 12.5 V/cm, and 15 V/cm) and water bath (WB) on microbial inactivation, physicochemical and sensory properties and microbial flora of pasteurized milk. Results indicated that OH with higher voltage could effectively inactivate microorganisms in milk, requiring less heating time and energy. Moreover, OH treatment at higher voltages could decelerate lipid oxidation and better maintain the sensory quality and essential amino acids content of milk. Additionally, all treatments significantly altered the microbial community, and during storage, the microbial community in milk treated with 10 V/cm and 12.5 V/cm OH remained relatively stable. OH treatments with voltage gradients exceeding 12.5 V/cm could effectively inactive microorganisms and maintain the quality attributes of milk.

A low-background and wash-free signal amplification F-CRISPR biosensor for sensitive quantitative and visible qualitative detection of Salmonella Typhimurium.

In traditional CRISPR-based biosensors, the cleavage-induced signal generation is insufficient because only a signals is generated at a CRISPR-induced cleavage. Herein, we developed an improved CRISPR/Cas12a-based biosensor with an enlarged signal generation which integrated the hybridization chain reaction (HCR) and low-background Förster Resonance Energy Transfer (FRET) signal output mode. The HCR with nucleic acid self-assembly capability was used as a signal carrier to load more signaling molecules. To get the best signal amplification, three different fluorescence signal output modes (fluorescence recovery, FRET and low-background FRET) generated by two fluoresceins, FAM and Cy5, were fully investigated and compared. The results indicated that the low-background FRET signal output mode with the strictest signal generation conditions yielded the highest signal-to-noise ratio (S/N) (19.17) and the most obvious fluorescence color change (from red to yellow). In optimal conditions, the proposed biosensor was successfully applied for Salmonella Typhimurium (S. Typhimurium) detection with 6 h (including 4 h for sample pre-treatment) from the initial target processing to the final detection result. The qualitative sensitivity, reliant on color changes, was 103 CFU/mL. The quantitative sensitivity, calculated by the fluorescence value, were 1.62 × 101 CFU/mL, 3.72 × 102 CFU/mL, and 8.71 × 102 CFU/mL in buffer solution, S. Typhimurium-spiked milk samples, and S.Typhimurium-spiked chicken samples, respectively. The excellent detection performance of the proposed biosensor endowed its great application potential in food and environment safety monitoring.

Bactericidal efficacy difference between air and nitrogen cold atmospheric plasma on Bacillus cereus: Inactivation mechanism of Gram-positive bacteria at the cellular and molecular level.

As an emerging food processing technology, cold atmospheric plasma (CAP) has attracted great attention in the field of microbial inactivation. Although CAP has been proven to effectively inactivate a variety of foodborne pathogens, there is less research on the inactivation of Bacillus cereus, and the exact inactivation mechanism is still unclear. Elucidating the inactivation mechanism will help to develop and optimize this sterilization method, with the prospective application in industrialized food production. This study aims to explore the bactericidal efficacy difference between air and nitrogen CAP on B. cereus, a typical Gram-positive bacterium, and reveals the inactivation mechanism of CAP at the cellular and molecular level, by observing the change of the cell membrane, cell morphological damage, intracellular antioxidant enzyme activity and cellular biomacromolecules changes. The results showed that both air CAP and nitrogen CAP could effectively inactivate B. cereus, which was due to the reactive oxygen and nitrogen species (RONS) generated by the plasma causing bacterial death. The damage pathways of CAP on Gram-positive bacteria could be explained by disrupting the bacterial cell membrane and cell morphology, disturbing the intracellular redox homeostasis, and destroying biomacromolecules in the cells. The differences in active species generated by the plasma were the main reason for the different bactericidal efficiencies of air CAP and nitrogen CAP, where air CAP producing RONS with stronger oxidative capacity in a shorter time. This study indicates that air CAP is an effective, inexpensive and green technology for B. cereus inactivation, providing a basis for industrial application in food processing.

Effects of mitochondria on postmortem meat quality: characteristic, isolation, energy metabolism, apoptosis and oxygen consumption.

Meat quality holds significant importance for both consumers and meat producers. Various factors influence meat quality, and among them, mitochondria play a crucial role. Recent studies have indicated that mitochondria can sustain their functions and viability for a certain duration in postmortem muscles. Consequently, mitochondria have an impact on oxygen consumption, energy metabolism, and apoptotic processes, which in turn affect myoglobin levels, oxidative stress, meat tenderness, fat oxidation, and protein oxidation. Ultimately, these factors influence the color, tenderness, and flavor of meat. However, there is a dearth of comprehensive summaries addressing the effects of mitochondria on postmortem muscle physiology and meat quality. Therefore, this review aims to describe the characteristics of muscle mitochondria and their potential influence on muscle. Additionally, a suitable method for isolating mitochondria is presented. Lastly, the review emphasizes the regulation of oxygen consumption, energy metabolism, and apoptosis by postmortem muscle mitochondria, and provides an overview of relevant research and recent advancements. The ultimate objective of this review is to elucidate the underlying mechanisms through which mitochondria impact meat quality.

Inactivation mechanisms of atmospheric pressure plasma jet on Bacillus cereus spores and its application on low-water activity foods.

Bacillus cereus spore is one of the most easily contaminated bacterial spores in low-water activity foods such as black pepper. Atmospheric-pressure plasma jet (APPJ) has emerged as an emerging and promising method for microbial inactivation in food processing. This study aimed to investigate the efficacy of APPJ in inactivating spores under various treatment parameters and to examine the resulting alterations in spore structures and internal membrane properties. Meanwhile, the practical application of APPJ for spore inactivation in black pepper was also evaluated. The results indicated that air-APPJ had superior spore inactivation capability compared to N2 and O2-APPJ. After 20 min of APPJ treatment (50 L/min, 800 W, and 10 cm), the reduction in spore count (>2 log CFU/g) was significantly greater than that achieved by heat treatment (80℃). The damage of inner membranes was considered as the major reason of the dried spore inactivation by APPJ treatment. Moreover, it achieved a reduction in spore count of > 1 log CFU/g on inoculated black pepper without significantly affecting its color and flavor. Although the antioxidant activity of black pepper was slightly reduced, the overall quality of the product was not considerably affected by plasma treatment. This study concluded that APPJ is an effective technique for spore inactivation, offering promising potential for application in the decontamination of low-water activity foods.

Effects of vegetable oil and ethylcellulose on the oleogel properties and its application in Harbin red sausage.

The effects of ethylcellulose (EC) concentration (6-12 %) and types of vegetable oil (sunflower, peanut, corn, and flaxseed oils) on the color, hardness, oil loss, lipid oxidation, and rheology property of oleogels were investigated in this study. Peanut oil (PO) oleogel was selected to replace pork fat partially in Harbin red sausage. Meanwhile, the fatty acid profile, texture, and sensory attributes of the reformulated sausages were analyzed. Oleogels formulated with higher EC concentration had higher brightness and hardness, a higher degree of lipid oxidation, and greater storage (G') and loss (G'') moduli. Oleogels formulated with PO had lower oil loss, flaxseed oil oleogel had higher hardness. Corn oil and PO oleogels had a lower level of lipid oxidation. The texture, lipid oxidation, and sensory attributes of the reformulated sausages, in which 10-30 % pork fat was replaced with PO oleogel, did not significantly differ from those without oleogel replacement. Meanwhile, the reformulated sausages had a healthier fatty acids profile and higher nutritional value.

Preparation of lemon essential oil nanoemulsion and its effect on the microbial community of pork patties.

The purpose of this study was to prepare an essential oil nanoemulsion and apply it to meat products to achieve antimicrobial effect. The nanoemulsion of lemon essential oil was produced by high power ultrasonication. The influences of ultrasonic power, ultrasonic time, and mass ratio of sodium caseinate to essential oil on the particle size were examined. The optimal conditions for preparing nanoemulsion were ultrasonic power 327 W, ultrasonic time 18 min, and sodium caseinate to essential oil mass ratio 4.23:1. Lower temperature was more conducive to the preservation of nanoemulsion. Compared to control and essential oil group, the nanoemulsion could decrease the total volatile basic nitrogen content and total bacterial colony in pork patties. When applied to pork patties, the nanoemulsion decreased the microbial diversity and inhibited the growth of a variety of microorganisms such as Bacillus, extending the storage time of pork patties. This study developed a novel and workable nanoemulsion for inhibiting bacteria of meat products.

Sublethally injured microorganisms in food processing and preservation: Quantification, formation, detection, resuscitation and adaption.

Sublethally injured state has been recognized as a survival strategy for microorganisms suffering from stressful environments. Injured cells fail to grow on selective media but can normally grow on nonselective media. Numerous microorganism species can form sublethal injury in various food matrices during processing and preservation with different techniques. Injury rate was commonly used to evaluate sublethal injury, but mathematical models for the quantification and interpretation of sublethally injured microbial cells still require further study. Injured cells can repair themselves and regain viability on selective media under favorable conditions when stress is removed. Conventional culture methods might underestimate microbial counts or present a false negative result due to the presence of injured cells. Although the structural and functional components may be affected, the injured cells pose a great threat to food safety. This work comprehensively reviewed the quantification, formation, detection, resuscitation and adaption of sublethally injured microbial cells. Food processing techniques, microbial species, strains and food matrix all significantly affect the formation of sublethally injured cells. Culture-based methods, molecular biological methods, fluorescent staining and infrared spectroscopy have been developed to detect the injured cells. Cell membrane is often repaired first during resuscitation of injured cells, meanwhile, temperature, pH, media and additives remarkably influence the resuscitation. The adaption of injured cells negatively affects the microbial inactivation during food processing.

Inactivation effects and mechanism of ohmic heating on Bacillus cereus.

The inactivation effects and mechanism of ohmic heating (OH) on Bacillus cereus ATCC 11778 were investigated in this study, conventional heating (CH) was also carried out and served as control. All OH treatments (10 V/cm 50 Hz, 10 V/cm 500 Hz, 5 V/cm 50 Hz and 5 V/cm 500 Hz) could achieve a comparable inactivation effect with CH, while OH treatments significantly shortened the processing time. OH treated cells exhibited significantly higher leakage of metal ions (Mg2+ and K+) and biomacromolecules (nucleic acids and proteins) than those treated with CH when bacterial suspensions were heated to the same temperature. Moreover, OH treatment caused more damage on membrane structure, greatly decreased the cell membrane potential and endogenous enzyme activity than that of CH. The results of this study indicated that OH is more efficient in the inactivation of bacteria.

Muscle fiber characteristics and apoptotic factor differences in beef Longissimus lumborum and Psoas major during early postmortem.

This study was conducted to study muscle fiber characteristics and apoptotic factor differences within 24 h postmortem of bovine longissimus lumborum (LL) and psoas major (PM). Compared to LL, PM had a higher proportion of type I fibers and lower proportion of type II fibers. PM also had higher ROS levels. For bcl-2 family proteins, anti-apoptotic BCL-2 level was lower and pro-apoptotic BAX level was higher in PM. For caspases, at 1 h postmortem, gene and protein expression level of caspase-3 and caspase-9 was higher in PM than that of LL. The levels of DNA damage apoptotic factors ABL1, AIF and ENDOG was higher in PM than in LL. The results suggested that apoptotic gene and protein expression were different in muscles with different fiber type composition. These findings provided insights into muscle fiber and apoptotic factor differences during early postmortem in bovine PM and LL.

Changes of mitochondrial lipid molecules, structure, cytochrome c and ROS of beef Longissimus lumborum and Psoas major during postmortem storage and their potential associations with beef quality.

This study investigated changes in mitochondrial lipid molecules and their potential associations with Longissimus lumborum (LL) and Psoas major (PM) quality during storage. A total of 1610 lipid species that matched with 36 lipid classes were identified from isolated mitochondria. PM had more key lipid molecules at storage d-1 including diacylglycerol and triacylglycerol (may play roles in membrane stability), phosphatidylethanolamine, acyl carnitine and cardiolipin (involved in energy metabolism), and cardiolipin and phosphatidylserine (important factors in apoptosis). Correspondingly, with extended storage time, mitochondrial structure, cytochrome c and reactive oxygen species (ROS) were changed, and muscle oxidation intensified. These changes have close associations with shear force and water holding capacity (WHC). Compared with LL, PM had higher content of lipid classes, more mitochondrial ROS, greater muscle oxidation, and lower shear force and WHC. These findings provided new insights into the effects of lipidome on mitochondrial structure, ROS and cytochrome c, and their potential associations with beef quality.

A CRISPR/Cas12a-mediated, DNA extraction and amplification-free, highly direct and rapid biosensor for Salmonella Typhimurium.

CRISPR/Cas-based biosensors were typically used for nucleic-acid targets detection and complex DNA extraction and amplification procedures were usually inevitable. Here, we report a CRISPR/Cas12a-mediated, DNA extraction and amplification-free, highly direct and rapid biosensor (abbreviated as "CATCHER") for Salmonella Typhimurium (S. Typhimurium) with a simple (3 steps) and fast (∼2 h) sensing workflow. Magnetic nanoparticle immobilized anti-S. Typhimurium antibody was worked as capture probe to capture the target and provide movable reaction interface. Colloidal gold labeled with anti-S. Typhimurium antibody and DNase I was used as detection probe to bridge the input target and output signal. First, in the presence of S. Typhimurium, an immuno-sandwich structure was formed. Second, DNase I in sandwich structure degraded the valid, complete activator DNA to invalid DNA fragments which can't trigger the trans-cleavage activity of Cas12a. Finally, the integrity of reporter DNA was preserved presenting a low fluorescence signal. Conversely, in the absence of S. Typhimurium, strong fluorescence recovery appeared owing to the cutting of reporter by activated Cas12a. Significantly, the proposed "CATCHER" showed satisfactory detection performance for S. Typhimurium with the limit of detection (LOD) of 7.9 × 101 CFU/mL in 0.01 M PBS and 6.31 × 103 CFU/mL in spiked chicken samples, providing a general platform for non-nucleic acid targets.

Employment of cold atmospheric plasma in chilled chicken breasts and the analysis of microbial diversity after the shelf-life storage.

Cold atmospheric plasma, featured as a non-thermal food processing technology has gained prominent attention for its practical application. The demand for chilled chicken has increased rapidly due to its nutritious value. However, chilled chicken often suffers from a short shelf life due to its susceptibility to spoilage. In this study, air- and nitrogen plasma were applied to investigate microbial inactivation effects and shelf-life extension of chilled chicken. Meanwhile, the microbial diversity of chicken breasts after air and nitrogen-CAP treatment and at the end of shelf-life was analyzed. Results illustrated that the inactivation effects of air plasma were more potent than that of nitrogen plasma, while nitrogen plasma could better maintain the sensory properties. Also, further investigation illustrated that air- and nitrogen-CAP could change the microbial community significantly. Both treatment time and working gases were essential experimental parameters that affected the microbial community composition.

Subcellular inactivation mechanisms of Pseudomonas aeruginosa treated by cold atmospheric plasma and application on chicken breasts.

As a novel microbe inactivation strategy, cold atmospheric plasma (CAP) technology has attracted great attractions in the past two decades. This study demonstrated that the CAP treatment was a robust inactivation approach for P. aeruginosa. Air and nitrogen-CAP achieved 100 % inactivation efficiency in 5 and 9 min, respectively. Furthermore, the inactivation mechanisms were explained by measuring several physicochemical indexes, including the characteristics of bacterial suspension, cell membrane integrity, cell viability, and the concentration of intracellular substances. The possible inactivation mechanisms might be that the RONS generated by air and nitrogen attacked the cell envelope, resulted in the leakage of intracellular substances; meanwhile, RONS also destroyed the intracellular anti-oxidative system, accelerated the oxidative stress and disrupted the intracellular redox homeostasis, subsequently the death of the cells. Moreover, the inactivation application in chicken breasts proved CAP could be a novel sanitizing process in practical industries.

Effect of Xanthan Gum, Kappa-Carrageenan, and Guar Gum on the Functional Characteristics of Egg White Liquid and Intermolecular Interaction Mechanism.

This study evaluated the effects of three polysaccharides, xanthan gum (XG), kappa-carrageenan (CA), and guar gum (GG), on the foaming and emulsifying properties of egg white liquid (EWL) and explored the intermolecular interactions and aggregation states in the initial polysaccharide−EWL complex. The results showed that the addition of XG and GG significantly improved the foaming stability of EWL on the one hand, from 66% to 78% and 69%, respectively (p < 0.05). On the other hand, the addition of XG and GG significantly improved the foam uniformity and density, and the average foam area decreased from 0.127 to 0.052 and 0.022 mm2, respectively (p < 0.05). The addition of XG and CA significantly improved the emulsification activity index (from 13.32 to 14.58 and 14.36 m2/mg, respectively, p < 0.05) and the emulsion stability index (from 50.89 to 53.62 and 52.18 min, respectively, p < 0.05), as well as the interfacial protein adsorption at the oil−water interface; it also reduced the creaming index. However, GG negatively affected these indicators. Furthermore, the electrostatic and hydrophobic interactions among molecules in EWL due to XG and the electrostatic, hydrogen bonding, and hydrophobic interactions among molecules in EWL due to CA ultimately led to the irregular aggregation of egg white proteins. Hydrophobic interactions and disulfide bonds between molecules in EWL−containing GG formed filamentous aggregations of egg white proteins. This work reveals that molecules in the polysaccharide−egg white complexes aggregate by interaction forces, which in turn have different effects on the foaming and emulsifying properties of egg white proteins.

Alteration of Mitochondrial Lipidome and Its Potential Effect on Apoptosis, Mitochondrial Reactive Oxygen Species Production, and Muscle Oxidation in Beef during Early Postmortem.

Lipid molecules are important participants in mitochondria-mediated apoptosis. This study explored the effect of mitochondrial lipids on mitochondria-mediated apoptosis, mitochondrial reactive oxygen species (ROS) production, and muscle oxidation of beef longissimus lumborum (LL, n = 6) and psoas major (PM, n = 6) during 24 h postmortem. A total of 432 lipid species matched with 21 lipid classes were identified. Remarkably, at 12 h postmortem, the levels of cardiolipin and phosphatidylserine in PM and ceramide, cardiolipin, phosphatidylserine, and sphingosine in LL increased significantly compared with 1 h postmortem, indicating that mitochondria-mediated apoptosis in beef muscle was accelerated during early postmortem. Moreover, PM had higher levels of cardiolipin and phosphatidylserine than LL, also suggesting a higher degree of apoptosis in PM during postmortem. Lipid molecules may assist the production of mitochondrial ROS and decrease the mitochondrial membrane potential (MMP) during postmortem apoptosis, resulting in muscle oxidation and the damage of antioxidant system. Notably, compared with LL, PM had higher abundance of apoptosis-related lipid molecules, a higher amount of ROS, faster diminishment in MMP, and then a higher degree of apoptosis. These findings provided new insights into the apoptosis and muscle biochemistry in beef during early postmortem.

Light activation of gold nanorods but not gold nanospheres enhance antibacterial effect through photodynamic and photothermal mechanisms.

Plasmonic nanomaterials of gold and silver have been reported to have antibacterial effect. In this study, three gold nanomaterials (NMs) of different aspect rations (Gold nanospheres (AuNSs, aspect ratio 1), and two gold nanorods (AuNRs636, aspect ratio 2.79; AuNRs772, aspect ratio 3.42)) and silver nanoparticles (AgNPs) were synthesized, characterized and the effect of incandescent light on their antibacterial properties were examined. Bacterial inactivation during photoinactivation of nanomaterials and antibacterial mechanisms (biotic ROS, membrane potential, membrane damage) were investigated using Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Salmonella enterica serovar Typhimurium, and methicillin-resistant S. aureus. The results indicated that AuNSs had no antibacterial activity in the tested concentration (0.49-250 µg/mL), while AuNR636 and AuNRs772 showed significant bactericidal effect on all tested bacteria. Notably, AuNRs636 presented higher antibacterial effect than AuNRs772, which could result from higher surface reactivity of AuNRs636 owing to higher dangling bonds. Further studies showed that AuNRs but not AuNSs generated hydroxyl radicals (·OH) (photodynamic effect) and photothermal effect when exposed to incandescent light. The combined photodynamic and photothermal effect resulted in bacterial inactivation through cell membrane damage, lowering of cell membrane potential and DNA degradation. In summary, this investigation showed that Au NRs but not Au NSs exhibit photodynamic and photothermal effects suggesting the potential of fabricating material surfaces with Au NRs for photoactivated bacterial inactivation.

The Use of Electronic Nose in the Quality Evaluation and Adulteration Identification of Beijing-You Chicken.

The objective of this study was to reveal the secrets of the unique meat characteristics of Beijing-you chicken (BJY) and to compare the difference of quality and flavor with Luhua chicken (LH) and Arbor Acres broiler (AA) at their typical market ages. The results showed the meat of BJY was richer in essential amino acids, arachidonic acid contents, inosine monophosphate (IMP), and guanosine monophosphate (GMP). The total fatty acid and unsaturated fatty acid contents of BJY chicken and LH chicken were lower than that of AA broilers, whereas the ratios of unsaturated fatty acids/saturated fatty acids (2.31) and polyunsaturated fatty acids/monounsaturated fatty acids (1.52) of BJY chicken were the highest. The electronic nose and SPME-GC/MS analysis confirmed the significant differences among these three chickens, and the variety and relative content of aldehydes might contribute to a richer flavor of BJY chicken. The meat characteristics of BJY were fully investigated and showed that BJY chicken might be favored among these three chicken breeds with the best flavor properties and the highest nutritional value. This study also provides an alternative way to identify BJY chicken from other chickens.

Effect of the Partial Substitution of Sodium Chloride on the Gel Properties and Flavor Quality of Unwashed Fish Mince Gels from Grass Carp.

Excessive salt is usually required to maintain good gel properties and quality characteristics for unwashed fish mince gels (UFMG). This study aimed to investigate the effects of partial sodium chloride substitution (30%) with different substitutes (potassium chloride, disodium inosine-5'-monophosphate, basil) on the gel and flavor properties of UFMG from Ctenopharyngodon idellus. The results indicated that the texture and gel strength of NK (30% NaCl was replaced with 30% KCl) were fairly similar to that of N group (NaCl only), and the whiteness had improved significantly (p < 0.05), while the product eventually yielded a certain bitter taste. The addition of disodium inosine-5'-monophosphate (DIMP) significantly (p < 0.05) increased the hardness, chewiness, buriedness degree of tryptophan and gel strength, decreased the content of α-helix structure in the gels, while less change occurred in gel whiteness and network structure. Basil significantly (p < 0.05) reduced the buriedness degree of tryptophan, gel strength and whiteness, and deteriorated the gel structure. Nevertheless, the addition of DIMP or basil reduced the bitterness induced by KCl and improved the overall acceptability scores of gels of the N group. Moreover, there was no distinct difference in moisture content and water-holding capacity between all groups. Therefore, replacing sodium chloride in UFMG with 25% potassium chloride and 5% DIMP may be an ideal sodium salt substitution strategy.

Recovery and virulence factors of sublethally injured Staphylococcus aureus after ohmic heating.

Ohmic heating (OH) is an alternative thermal processing technique, which is widely used to pasteurize or sterilize food. However, sublethally injured Staphylococcus aureus induced by OH is a great concern to food safety. The recovery of injured S. aureus by OH and virulence factor changes during recovery were investigated in this study. The liquid media (phosphate-buffered saline, buffered peptone water and nutrient broth (NB)), temperature (4, 25 and 37 °C) and pH (6.0, 7.2 and 8.0) influenced the recovery rate and the injured cells completely repaired in NB at 37 °C, pH 7.2 with the shortest time of 2 h. The biofilm formation ability, mannitol fermentation, hemolysis, and coagulase activities decreased in injured S. aureus and recovered during repair process. Quantitative real-time PCR showed the expression of sek, clfB and lukH involved in virulence factors increased during recovery. The results indicated that the virulence factors of injured S. aureus recovered after repair.