Contribution of mono- and co-culture of Pseudomonas paralactis, Acinetobacter MN21 and Stenotrophomonas maltophilia to the spoilage of chill-stored lamb.

Exploring the contribution of common microorganisms to spoilage is of great significance in inhibiting spoilage in lamb. This work investigated the extent of protein degradation and profile changes of free amino acids (FAAs), free fatty acids (FFAs) and volatile organic compounds (VOCs) in lamb caused by single- and co-culture of the common aerobic spoilage bacteria, P. paralactis, Ac. MN21 and S. maltophilia. Meanwhile, some key VOCs produced by the three bacteria during lamb spoilage were also screened by orthogonal partial least square discriminant analysis and difference value in VOCs content between inoculated groups and sterile group. Lamb inoculated with P. paralactis had the higher total viable counts, pH, total volatile base nitrogen and TCA-soluble peptides than those with the other two bacteria. Some FAAs and FFAs could be uniquely degraded by P. paralactis but not Ac. MN21 and S. maltophilia, such as Arg, Glu, C15:0, C18:0 and C18:1n9t. Co-culture of the three bacteria significantly promoted the overall spoilage, including bacterial growth, proteolysis and lipolysis. Key VOCs produced by P. paralactis were 2, 3-octanedione, those by Ac. MN21 were 1-octanol, octanal, hexanoic acid, 1-pentanol and hexanoic acid methyl ester, and that by S. maltophilia were hexanoic acid. The production of extensive key-VOCs was significantly and negatively correlated with C20:0, C23:0 and C18:ln9t degradation. This study can provide a basis for inhibiting common spoilage bacteria and promoting high-quality processing of fresh lamb.

Acetylation and Phosphorylation Regulate the Role of Pyruvate Kinase as a Glycolytic Enzyme or a Protein Kinase in Lamb.

Protein post-translational modifications (PTMs) play an essential role in meat quality development. However, the effect of specific PTM sites on meat proteins has not been investigated yet. The characteristics of pyruvate kinase M (PKM) were found to exhibit a close correlation with final meat quality, and thus, serine 99 (S99) and lysine 137 (K137) in PKM were mutated to study their effect on PKM function. The structural and functional properties of five lamb PKM variants, including wild-type PKM (wtPKM), PKM_S99D (S99 phosphorylation), PKM_S99A (PKM S99 dephosphorylation), PKM_K137Q (PKM K137 acetylation), and PKM_K137R (PKM K137 deacetylation), were evaluated. The results showed that the secondary structure, tertiary structure, and polymer formation were affected among different PKM variants. In addition, the glycolytic activity of PKM_K137Q was decreased because of its weakened binding with phosphoenolpyruvate. In the PKM_K137R variant, the actin phosphorylation level exhibited a decrease, suggesting a low kinase activity of PKM_K137R. The results of molecular simulation showed a 42% reduction in the interface area between PKM_K137R and actin, in contrast to wtPKM and actin. These findings are significant for revealing the mechanism of how PTMs regulate PKM function and provide a theoretical foundation for the development of precise meat quality preservation technology.

Fluctuation of flavor quality in roasted duck: The consequences of raw duck preform's repetitive freeze-thawing.

This study aimed to investigate the changes in flavor quality of roasted duck during repetitive freeze-thawing (FT, -20 ℃ for 24 h, then at 4 ℃ for 24 h for five cycles) of raw duck preforms. HS-SPME/GC-MS analysis showed that more than thirty volatile flavor compounds identified in roasted ducks fluctuated with freeze-thawing of raw duck preforms, while hexanal, nonanal, 1-octen-3-ol, and acetone could as potential flavor markers. Compared with the unfrozen raw duck preforms (FT-0), repetitive freeze-thawing increased the protein/lipid oxidation and cross-linking of raw duck preforms by maintaining the higher carbonyl contents (1.40 âˆ¼ 3.30 nmol/mg), 2-thiobarbituric acid reactive substances (0.25 âˆ¼ 0.51 mg/kg), schiff bases and disulfide bond (19.65 âˆ¼ 30.65 µmol/g), but lower total sulfhydryl (73.37 âˆ¼ 88.94 µmol/g) and tryptophan fluorescence intensity. Moreover, A lower protein band intensity and a transformation from α-helixes to ß-sheets and random coils were observed in FT-3 âˆ¼ FT-5. The obtained results indicated that multiple freeze-thawing (more than two cycles) of raw duck preforms could be detrimental to the flavor quality of the roasted duck due to excessive oxidation and degradation.

Research progress of chilled meat freshness detection based on nanozyme sensing systems.

It is important to develop rapid, accurate, and portable technologies for detecting the freshness of chilled meat to meet the current demands of meat industry. This report introduces freshness indicators for monitoring the freshness changes of chilled meat, and systematically analyzes the current status of existing detection technologies which focus on the feasibility of using nanozyme for meat freshness sensing detection. Furthermore, it examines the limitations and foresees the future development trends of utilizing current nanozyme sensing systems in evaluating chilled meat freshness. Harmful chemicals are produced by food spoilage degradation, including biogenic amines, volatile amines, hydrogen sulfide, and xanthine, which have become new freshness indicators to evaluate the freshness of chilled meat. The recognition mechanisms are clarified based on the special chemical reaction with nanozyme or directly inducting the enzyme-like catalytic activity of nanozyme.

Development of a rhodamine-based fluorescent probe for ATP detection for potential applications in meat freshness assessment.

Adenosine triphosphate (ATP) plays a vital role in physiological processes and is an essential indicator of microbial content in food. Herein, a new sensitive, rapid and water-soluble probe for ATP detection was developed. Rhodamine B and pentaethylenehexamine were employed to design and synthesise the probe rhodamine-pentaethylenehexamine (RP) for selective ATP detection. The synthesised probe RP was characterized using Fourier transform infrared, NMR and dynamic light scattering size distributions. Upon the addition of ATP, the probe exhibited a distinct change in fluorescence intensity, with fluorescence emission at 580 nm. A linear relationship was observed between fluorescence intensity and ATP concentrations at 0-50 µmol/L, with a limit of detection of 10.97 × 10-9 mol/L. The results of the zeta potential and molecular dynamics simulation demonstrated that the detection mechanism of the probe RP is associated with the electrostatic adsorption interaction between the multi-positively charged sites of RP and the negatively charged triphosphate structure of ATP. Our study provides new insights into improving charge site identification in small molecule detection. Furthermore, the successful detection of ATP on meat surfaces indicates that RP has the potential to assess meat freshness.

Conformational changes induced by selected flavor compounds from spices regulate the binding ability of myofibrillar proteins to aldehyde compounds.

Interactions among flavor compounds from spices (FCS) and myofibrillar proteins (MP) were investigated. Fluorescence and Fourier transform infrared spectroscopy showed that hydrogen bonding and hydrophobic interactions were the main binding forces between FCS and MP. The FCS increased the particle size and SH content of MP and caused a reduction of zeta potential from -5.23 to -6.50 mV. Furthermore, FCS could modify the binding ability of MP and aldehydes. Eugenol reduced the ability of MP to bond with aldehydes by 22.70-47.87 %. Molecular dynamics simulations demonstrated that eugenol may combat nonanal to attain binding site of amino acid residue (PHE165) and induce protein conformational changes. Electrostatic interactions and van der Waals forces within myosin-nonanal may be disrupted by these alterations, which could reduce stability of complex and cause release of nonanal. This study could provide new insights into regulating the ability of proteins to release and hold flavors.

Simultaneous determination of advanced glycation end products and heterocyclic amines in roast/grilled meat by UPLC-MS/MS.

Advanced glycation end products (AGEs) and heterocyclic amines (HAs) are main harmful Maillard reaction products of meat products. Simultaneous quantification of both with high sensitivity, selectivity and accuracy remains a major challenge due to inconsistencies in their pre-treatment and instrumental methods and the different polarity of AGEs and HAs. We developed a method for the simultaneous determination of AGEs and HAs in roast/grilled meat by ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) using dynamic multiple reaction monitoring (D-MRM). The instrument parameters and pre-treatment method were optimized to achieve reasonably good separation and high response for the 11 target analytes within 8 min. From 10 to 200 ng/mL, the limits of detection (LODs) and limits of quantitation (LOQs) ranged from 0.3 to 5.5 µg/L and 0.9 to 6.3 µg/L, respectively, and the correlation coefficient (R2) was >0.99. It was acceptable to recoveries, standard deviations (RSDs), and matrix effects. Six types of roast/grilled meat samples were then tested using the developed method.

A hydrogel-based ratiometric fluorescent sensor relying on rhodamine B labelled AIE-featured hyperbranched poly(amido amine) for heparin detection.

The fluorescent flexible sensor for point-of-care quantification of clinical anticoagulant drug, Heparin (Hep), is still an urgent need of breakthrough. In this research, a hyperbranched poly(amido amine) (HPA) was decorated with tetraphenylethene (TPE) and Rhodamine B (RhB), constructing a ratiometric fluorescent sensor (TR-HPA) for Hep. When the sensor was exposed to Hep, the TPE units within the probe skeleton would aggregate, resulting in an increasing fluorescent emission at 483 nm. The 580 nm of fluorescence came from RhB enhance, simultaneously, due to the fluorescence resonance energy transfer. As a result, there are two good linear correlation between the fluorescence emission ratio (E483/E580) of TR-HPA and the Hep concentration over a range of 0-1.0 µM, with a low limit of detection of 3.0 nM. Furthermore, we incorporate the TR-HPA probe into a polyvinyl alcohol (PVA) hydrogel matrix to create a flexible fluorescent sensing system platform, denoted as TR-HPA/PVA. This approach offers a straightforward visual detection method by causing a fluorescence color change from pink to blue when trace amounts of Hep are present. The hydrogel-based fluorescent sensor streamlines the detection procedures for Hep in biomedical applications. It shows great potential in rapid and point-of-care human blood clotting condition monitoring, making it suitable for next-generation wearable medical devices.

Insights into flavor formation of braised chicken: Based on E-nose, GC-MS, GC-IMS, and UPLC-Q-Exactive-MS/MS.

Effects of braising duration on volatile organic compounds (VOCs) and lipids in chicken were investigated. Aroma profiles identified by an electronic nose were effective in differentiating braising stages. During braising process, a total of 25 key VOCs were detected in braised chicken, and sample braised for 210 min exhibited the highest level of key VOCs. Additionally, a gas chromatography mass spectrometry fingerprint was established to evaluate the distribution of VOCs throughout the braising process. Partial least square discriminant analysis indicated that 2-heptanone, 3-methyl-2-butanone, octanal, nonanal, butanal, (E)-2-pentenal, 1-octen-3-ol, 1-hexanol, pentanal, hexanal, and 1-pentanol significantly affected flavor characteristics of braised chicken. Furthermore, 88 differential lipids were screened, and glycerolipids metabolic was found to be main metabolic pathway during braising process. Triglycerides (TG) and phosphatidyl ethanolamine (PE), such as TG (16:0/18:1/18:2), TG (18:0/18:1/18:2), TG (18:1/18:2/18:3), TG (18:1/18:1/18:2), PE (O-18:2/18:2), PE(O-18:2/18:1), and TG (16:0/16:1/18:2), played a vital role in the generation of VOCs.

Zein and tannic acid hybrid particles improving physical stability, controlled release properties, and antimicrobial activity of cinnamon essential oil loaded Pickering emulsions.

Pickering emulsion loading essential oil has demonstrated a promising strategy as delivery system in food preservation, but localization in stability and antimicrobial activity limits application. In this study, Pickering emulsions co-loaded with tannic acid and cinnamon essential oil (ZTC) have been developed based on zein and tannic acid complexes (ZT) mediated interfacial engineering. Fourier transform infrared, fluorescence spectroscopy, and molecular docking results indicated tannic acid altered the structural of zein. Interfacial tension results indicated that tannic acid accelerated the adsorbed speed of zein particles by decreased interfacial tension (11.99-9.96 mN/m). ZT5 formed a viscoelastic and dense layer in oil-water interface than that for other ZTs, which improved stability and control release performance of ZTC. Furthermore, the ZTC showed an effective antimicrobial activity against spoilage organisms Pseudomonad paralactis MN10 and Lactobacillus sakei VMR17. These findings provide new insight for developing co-loaded multiple antimicrobial agents within Pickering emulsion as a delivery system.

Effects of different cold chain logistics modes on the quality and bacterial community succession of fresh pork.

The connection and temperature control of cold chain links are key to ensuring meat quality. Considering the practical production of cold chain logistics in China, this study investigated the impact of various cold chain logistics modes (including warehousing, transportation, and sales conditions) at different temperature settings (7 °C, 4 °C, and - 1 °C), modeled in the laboratory, on the quality and bacterial community succession of pork. The pork quality was evaluated by pH, water holding capacity, total volatile basic nitrogen (TVB-N), total viable count (TVC) and myowater status. Among the different cold chain logistics modes, the LL1 (samples being warehoused and transported at 4 °C for 96 h and sold at -1 °C) and the SL1 (samples being warehoused and transported at 4 °C for 30 h and sold at -1 °C) modes were suitable for inter-provincial and intra-provincial transportation due to their long shelf life (> 14 days), respectively. The bacterial community succession of pork in different cold chain logistics modes was accessed by high-throughput sequencing. The results indicated that the cold chain logistics modes had affected the bacterial community, with Latilactobacillus being the dominant bacteria in the LL1 mode and SL1 mode during spoilage. The study revealed that the entire or partial process supercooling treatment (-1 °C) during the cold chain logistics process could effectively preserve the meat quality, supporting the high-quality development of the fresh meat cold chain logistics.

Investigating the impact of lipid molecules and heat transfer on aroma compound formation and binding in roasted chicken skin: A UHPLC-HRMS and GC-O-MS study.

The UHPLCHRMS and Gas Chromatography-Olfactometry-Mass Spectrometry (GC-O-MS) techniques were applied to investigate effects of lipid molecules and heat transfer on the generation of aroma compounds in roasted chicken skin. Nineteen odorants were identified as most important aroma contributors based on odor activity values (OAVs) exceeding 1. Lipidomic analysis identified 3926 lipids in the samples, in which triglycerides (TG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), and ceramide (Cer) had a contribution of 20.63%, 12.46%, 11.95%, and 11.39%, respectively. Furthermore, it was observed that PS(18:3e_22:5) and TG(18:0_18:1_18:1) serve as significant chemical markers for distinguishing chicken skin during the roasting (p < 0.05). TGs, notably TG(16:1_18:1_18:2) and TG(18:1_18:2_18:2), were postulated as key retainers for binding crucial aroma compounds. Meanwhile, PC, PE, and Cer played pivotal roles in aroma compound formation. Additionally, higher thermal conductivity and reduced thermal diffusivity significantly contributed to the formation of key odorants.

Improving the Edible and Nutritional Quality of Roasted Duck Breasts through Variable Pressure Salting: Implications for Protein Anabolism and Digestion in Rats.

Variable pressure salting (VPS) is considered a novel salting approach to improve meat quality. This study aimed to investigate the effects of roasted duck's edible and nutritional quality after VPS through serum biochemical indicators and in vivo digestion properties in rats. The results show that roasted duck after VPS led to an increase in the total protein content (57.24 g/L) and blood glucose levels (6.87 mmol/L), as well as a decrease in the blood urea nitrogen content (11.81 mmol/L), in rats. Compared to rats fed base diets and roasted duck after static wet salting (SWS), those ingesting roasted duck after VPS exhibited higher values of apparent protein digestibility (51.24%), pepsin activity (2.40 U/mg), and trypsin activity (389.80 U/mg). Furthermore, VPS treatment improved the textural properties and microstructure of duck breasts shown by a higher immobilized water relaxation area and more ordered protein structures (α-helixes and ß-sheets). These improvements enhanced the protein anabolism capacity and in vivo digestion properties in rats. Therefore, VPS represents a beneficial salting method for promoting effective digestion and absorption in rats.

Characterization of key aroma compounds in roasted chicken using SPME, SAFE, GC-O, GC-MS, AEDA, OAV, recombination-omission tests, and sensory evaluation.

Aroma compounds in the roasted breasts, thighs and skins of chicken were isolated by solvent-assisted flavor evaporation (SAFE), quantitated by gas chromatography-olfactometry-mass (GC-O-MS), analyzed by aroma extract dilution analysis (AEDA), and determined by recombination-omission tests and sensory evaluation. Forty-seven aroma compounds in total, including aldehydes, ketones, furans, pyrazines, and furanones, were selected by AEDA. Twenty-five compounds were selected as pivotal odorants (Odor Activity Value, OAV ≥ 1). Twenty aroma compounds significantly were identified by recombination and omission experiments. Anethole (fennel odor) was the highest OAV (＞ 1843). Hexanal (grassy) and (E, E)-2,4-decadienal (meaty) were the most abundant aldehydes identified in roasted chicken. 1-octen-3-ol (mushroom), methanethiol (cabbage) and dimethyl trisulfide (areca, sulfur) were considered the key compounds of the breast and thighs of roasted chicken. Notably, furanone and pyrazines, 4-hydroxy-5-methyl-3(2H)-furanone (caramel, sweet and burning odor), 3-ethyl-2,5-dimethylpyrazine (nutty, toasty) and 2,3-dimethyl-5-ethylpyrazine (nutty, toasty) had the most significant effect on roasted chicken odor, especially in the skin.

Effects of phosphoglycerate kinase 1 and pyruvate kinase M2 on metabolism and physiochemical changes in postmortem muscle.

The objective of this work was to investigate the influence of phosphoglycerate kinase-1 (PGK1) and pyruvate kinase-M2 (PKM2) activity on glycolysis, myofibrillar proteins, calpain system, and apoptosis pathways of postmortem muscle. The activity of PGK1 and PKM2 was regulated by their inhibitors and activators to construct the postmortem glycolysis vitro model and then incubated at 4 °C for 24 h. The results showed that compared to PGK1 and PKM2 inhibitors groups, the addition of PGK1 and PKM2 activators could accelerate glycogen consumption, ATP and lactate production, while declining pH value. Moreover, the addition of PGK1 and PKM2 activators could increase desmin degradation, µ-calpain activity, and caspase-3 abundance. Interestingly, troponin-T degradation was significantly increased both in PKM2 inhibitor and activator groups. It was suggested that PGK1 and PKM2 might be used as robust indicators to regulate meat quality by affecting the glycolysis, myofibrillar proteins, µ-calpain and apoptosis pathways in postmortem muscle.

Changes in physicochemical properties and formation process of colloidal nanoparticles (CNPs) during the lamb soup stewing.

Colloidal nanoparticles (CNPs), as carriers of nutrients, naturally exist in food or form during cooking. In this study, the colloidal properties, structures, rheological properties, and chemical composition location of CNPs were analyzed during 15 min to 5 h lamb soup stewing. With the increasing stewing time, the particle size and absolute value of the zeta potential of CNPs increased, indicating that CNPs became more stable. As the stewing time increased, the blue-shifted Fourier transform infrared spectroscopy absorption peaks and the red-shifted fluorescence spectroscopy absorption peaks certificated the structural changes in CNPs. And α-helix and ß-turn content decreased, while ß-sheet and random coil content increased in processing, potentially resulting in the opening CNPs structures. In addition, our findings revealed that proteins were encapsulated within the lipids in the inner part, while carbohydrates were dispersed in the outermost layers of the CNPs with a phospholipid bilayer.

Comparative effects of phosphorylation and acetylation on glycolysis and myofibrillar proteins degradation in postmortem muscle.

The study investigated the different effects between protein phosphorylation and acetylation on glycolytic enzyme activity and myofibrillar protein degradation. Lamb longissimus thoracis lumborum muscles were homogenized and then inhibitors were added for incubation at 4 °C. Phosphatase inhibitor was added to produce a high phosphorylation level (PI group) and lysine deacetylase inhibitor was added to produce a high acetylation level (DI group). The lactate and ATP content in the PI group was inhibited compared with that in the DI group (P < 0.05). Phosphofructokinase (PFK) activity was negatively related with the phosphorylation level and was positively related with the acetylation level in the DI group (P < 0.05). The degradation of troponin T and desmin of the DI group were restrained when compared to that in the PI group (P < 0.05). Compared with initial PFK and desmin, the simulation of phosphorylation and acetylation of PFK and desmin showed different electrostatic potential at the surface and a more unstable structure. The phosphorylation level of the DI group was increased, suggesting that the changes of protein acetylation altered protein phosphorylation. In conclusion, compared with protein phosphorylation, protein acetylation had a greater effect on promoting glycolysis and inhibiting protein degradation.

Effects of different electrostatic field intensities assisted controlled freezing point storage on water holding capacity of fresh meat during the early postmortem period.

In this study, the effect of different intensity electrostatic fields on the water holding capacity (WHC) of fresh meat during the early postmortem period in controlled freezing point storage (CFPS) were investigated. Significantly lower cooking loss were found in low voltage electrostatic field (LVEF) and high voltage electrostatic field (HVEF) compared to the control group (CK) (p < 0.05). The myofibril fragmentation index and microstructure results suggested that the sample under HVEF treatment remained relatively intact. It has been revealed that the changes in actomyosin properties under electrostatic field treatment groups were due to the combination and dissociation of actomyosin binding into myofilament concentration, which consequently affects the muscle WHC. The study further demonstrated that the electrostatic field, especially HVEF, might increase the WHC of fresh meat by affecting the distribution of water molecules and physiochemical properties of actomyosin during the early postmortem period.

The effect of modified atmosphere packaging at an early postmortem stage on lamb meat quality during subsequent aging.

This study explores the influence of modified atmosphere packaging (MAP) on fresh lamb meat quality with respect to gas concentration, rigor state, and post-mortem aging time. A comparison was done for the quality characteristics of lamb Longissimus thoracis lumborum chops that had been packaged separately in air, 75%O2  + 25%CO2 MAP or 50%O2  + 50%CO2 MAP at 1, 6, and 24 h post-mortem and then stored for 6, 12, 24, 72, and 144 h post-mortem, and the quality of lamb chops had been evaluated at each post-mortem period separately. Chops packaged at 1 and 6 h post-mortem in MAP had reduced pH decline, less purge loss, and enhanced redness at early post-mortem storage times. Lamb color stability was evidently greater in 75%O2  + 25%CO2 MAP than in 50%O2  + 50%CO2 MAP during the early storage period when a* and R630/R580 were taken into account. Shear force values were lowest in lambs packaged at 1 h post-mortem with 75%O2  + 25%CO2 MAP at 12 h post-mortem and then increased until 72 h post-mortem, suggesting that rigor has been delayed by such a high O2 MAP. Thus, fresh lamb quality was maintained most effectively when packaged at 1 h post-mortem in 75%O2  + 25%CO2 MAP for consumption at 12 h post-mortem. The exact mechanisms and optimization of MAP based on Chinese retail conditions should be considered in future studies. PRACTICAL APPLICATION: In this study, three slaughter patterns in the meat industry involving boning immediately after dressing (hot-boning) and chilling for a short period (warm-boning) or overnight (cold-boning) are considered, as well as the behavior of non-immediate consumption after purchase. Modified atmosphere packaging provides an effective preservation of early post-mortem muscles with enhanced color stability, water holding capacity, and texture during refrigerated storage. This could provide new insights into how to process lamb muscles in the early post-mortem period to improve and stabilize lamb quality.

Insights into the interactions between etheric compounds and myofibrillar proteins using multi-spectroscopy, molecular docking, and molecular dynamics simulation.

This study aimed to examine how the addition of etheric compounds (EC) affects the characteristics of myofibrillar proteins (MP) and to understand underlying interaction mechanisms. Fourier transform infrared spectroscopy confirmed that the EC-MP complex was formed through hydrogen bonding. The addition of EC resulted in an increase in the α-helix content and a decrease in the ß-sheet content of MP, which would promote the protein unfolding. The unfolding of MP led to aggregation and formation of larger and non-uniform particles. As a result, the exposure of negative charge on the MP surface was enhanced, and zeta potential was decreased from -5.33 mV to -7.45 mV. Moreover, the EC-induced modification of MP conformation resulted in a less rigid three-dimensional network structure of MP gel and enhanced the discharge of aldehyde compounds (C > 6). Moreover, the rheological characteristics of MP were enhanced by the suppression of protein-protein interactions due to the MP unfolding. Molecular dynamics simulations revealed that anethole reduced the binding capacity of myosin to decanal by raising its binding energy from -22.22 kcal/mol to -19.38 kcal/mol. In the meantime, anethole competed for the amino acid residue (PHE165) where myosin connects to decanal. This caused the hydrogen bonds and hydrophobic contacts between the two molecules to dissolve, altering myosin's conformation and releasing decanal. The results might be useful in predicting and controlling the ability of proteins to release and hold onto flavors.