The Effect of the PhoP/PhoQ System on the Regulation of Multi-Stress Adaptation Induced by Acid Stress in Salmonella Typhimurium.

Acidic stress in beef cattle slaughtering abattoirs can induce the acid adaptation response of in-plant contaminated Salmonella. This may further lead to multiple resistance responses threatening public health. Therefore, the acid, heat, osmotic and antibiotic resistances of Salmonella typhimurium (ATCC14028) were evaluated after a 90 min adaption in a pH = 5.4 "mild acid" Luria-Bertani medium. Differences in such resistances were also determined between the ∆phoP mutant and wild-type Salmonella strains to confirm the contribution of the PhoP/PhoQ system. The transcriptomic differences between the acid-adapted and ∆phoP strain were compared to explore the role of the PhoP/Q two-component system in regulating multi-stress resistance. Acid adaptation was found to increase the viability of Salmonella to lethal acid, heat and hyperosmotic treatments. In particular, acid adaptation significantly increased the resistance of Salmonella typhimurium to Polymyxin B, and such resistance can last for 21 days when the adapted strain was stored in meat extract medium at 4 °C. Transcriptomics analysis revealed 178 up-regulated and 274 down-regulated genes in the ∆phoP strain. The Salmonella infection, cationic antimicrobial peptide (CAMP) resistance, quorum sensing and two-component system pathways were down-regulated, while the bacterial tricarboxylic acid cycle pathways were up-regulated. Transcriptomics and RT-qPCR analyses revealed that the deletion of the phoP gene resulted in the down-regulation of the expression of genes related to lipid A modification and efflux pumps. These changes in the gene expression result in the change in net negative charge and the mobility of the cell membrane, resulting in enhanced CAMP resistance. The confirmation of multiple stress resistance under acid adaptation and the transcriptomic study in the current study may provide valuable information for the control of multiple stress resistance and meat safety.

Effects and mechanism of sub-freezing storage on water holding capacity and tenderness of beef.

In order to explore the effect of sub-freezing storage on water holding capacity and tenderness of beef, four treatments were compared in this study: sub-freezing (-7 °C) fast sub-freezing (-38 °C until the core temperature achieved to -7 °C), superchilling (-1 °C) and fast frozen (-38 °C until the core temperature achieved to -18 °C) with the latter two treatments serving as the controls. The differences in muscle fiber structure, water distribution, protein oxidation and cytoskeletal protein degradation were studied. The results demonstrated that compared with other treatments, the fast sub-freezing treatment resulted in less structural damage to the muscle fibers and had better water holding capacity. Both sub-freezing and fast sub-freezing treatments inhibited protein oxidation compared with superchilling, but the former treatment's level of protein oxidation was higher than that in fast sub-freezing treatment during long-term storage (42 weeks). In addition, the structural proteins in the sub-freezing and fast sub-freezing treatments underwent faster degradation during long-term storage and therefore the meat was more tender compared with the fast frozen treatment. The results indicate that the fast sub-freezing treatment can be potentially applied in beef storage.

LC-MS-based metabolomics reveals metabolite dynamic changes of beef after superchilling early post-mortem.

To explore the underlying mechanisms by which superchilling (SC, -3 °C within 5 h of slaughter) improves beef tenderness, an untargeted metabolomics strategy was employed. M. Longissimus lumborum (LL) muscles from twelve beef carcasses were assigned to either SC or very fast chilling (VFC, 0 °C within 5 h of slaughter) treatments, with conventional chilling (CC, 0 âˆ¼ 4 °C until 24 h post-mortem) serving as the control (6 per group). Biochemical properties and metabolites were investigated during the early post-mortem period. The results showed that the degradation of µ-calpain and caspase 3 occurred earlier in SC treated sample, which might be attributed to the accelerated accumulation of free Ca2+. The metabolomic profiles of samples from the SC and CC treatments were clearly distinguished based on partial least squares-discriminant analysis (PLS-DA) at each time point. It is noteworthy that more IMP and 4-hydroxyproline were found in the comparison between SC and CC treatments. According to the results of metabolic pathways analysis and the correlation analysis between traits related to tenderness and metabolites with significant differences (SC vs. CC), it can be suggested that the tenderization effect of the SC treatment may be related to the alteration of arginine and proline metabolism, and purine metabolism in the early post-mortem phase.

Airborne signals of Pseudomonas fluorescens modulate swimming motility and biofilm formation of Listeria monocytogenes in a contactless coculture system.

Bacterial volatile compounds (BVCs) facilitate interspecies communication in socio-microbiology across physical barriers, thereby influencing interactions between diverse species. The impact of BVCs emitted from Pseudomonas on the biofilm formation characteristics of Listeria monocytogenes within the same ecological niche has been scarcely investigated under practical conditions of food processing. The objective of this study was to explore the motility and biofilm formation characteristics of L. monocytogenes under the impact of Pseudomonas BVCs. It was revealed that BVCs of P. fluorescens, P. lundensis, and P. fragi significantly promoted swimming motility of L. monocytogenes (P < 0.05). As evidenced by crystal violet staining, the L. monocytogenes biofilms reached a maximum OD570 value of approximately 3.78 at 4 d, which was 0.65 units markedly higher than that of the control group (P < 0.05). Despite a decrease in adherent cells of L. monocytogenes biofilms among the BVCs groups, there was a remarkable increase in the abundance of extracellular polysaccharides and proteins with 3.58 and 4.90 µg/cm2, respectively (P < 0.05), contributing to more compact matrix architectures, which suggested that the BVCs of P. fluorescens enhanced L. monocytogenes biofilm formation through promoting the secretion of extracellular polymers. Moreover, the prominent up-regulated expression of virulence genes further revealed the positive regulation of L. monocytogenes under the influence of BVCs. Additionally, the presence of BVCs significantly elevated the pH and TVB-N levels in both the swimming medium and biofilm broth, thereby exhibiting a strong positive correlation with increased motility and biofilm formation of L. monocytogenes. It highlighted the crucial signaling regulatory role of BVCs in bacterial interactions, while also emphasizing the potential food safety risk associated with the hitchhiking behavior of L. monocytogenes, thereby shedding light on advancements in control strategies for food processing.

Exploration of the shelf-life difference between chilled beef and pork with similar initial levels of bacterial contamination.

This study compared the shelf-life of beef and pork longissimus lumborum muscles (loins) that had the same initial bacterial loads and were held under the same chilled storage conditions. To identify the underlying pathways, comparisons were conducted from the perspective of the spoilage indicators; protease/lipase activity, and the volatile organic compounds (VOC) generated over 28 d of chilled storage. The initial total viable microbial count (TVC) on Day 0 for both type of meat was 4.3 log10 CFU/g. It was found that the TVC of beef and pork did not differ throughout the total chilled storage period and both ultimately exceeded 7 log10 CFU/g after 28 d. Based on total volatile basic nitrogen (TVB-N) guidelines, pork was spoilt after 21 d of chilled storage and therefore 7 d earlier than beef. Changes in the concentration of VOC spoilage biomarkers, including 1-octen-3-ol, 1-octanol, nonanal, and others, confirmed that pork had a shorter shelf-life than beef. An important reason for the difference in shelf-life between the two types of meat was that pork had a higher protease activity, although the beef had higher levels of total lipase activity. These findings help us understand the differences in the spoilage process of raw meat from different species and explore specific measures to control the spoilage of beef or pork.

Effects of CO2 on the physicochemical, microbial, and sensory properties of pork patties packaged under optimized O2 levels.

The storage quality characteristics of fresh pork patties were investigated under 80% O2 modified atmosphere packaging (MAP80:20 = 80% O2/20% CO2) and 40% O2 MAP with various CO2 levels (MAP40:20 = 40% O2/20% CO2/40% N2; MAP40:40 = 40% O2/40% CO2/20% N2; MAP40:60 = 40% O2/60% CO2). Packaged patties were stored for 16 days at 4 °C to monitor their physicochemical (pH, instrumental color, oxidative stability, and fatty acid profile), microbial, and sensorial changes. Results suggested that decreasing O2 levels from 80% to 40% significantly inhibited the lipid oxidation of patties but led to a lower (P < 0.05) color stability. Elevating CO2 levels from 20% to 60% in combination with 40% O2 significantly suppressed bacterial growth and total volatile basic nitrogen production, and thus rendered patties with a better sensory quality and a similar meat color to 80% O2. However, increased CO2 levels promoted lipid oxidation through reducing the antioxidant capacity of patties, which was attributed to a CO2-induced reduction in superoxide dismutase and glutathione peroxidase activities during storage rather than a pH reduction or changes in fatty acid composition. Overall, 40% O2/40% CO2/20% N2 is a realistic alternative for pork patties to improve meat quality and extend the shelf-life to over 16 days.

Study of the transfer of Shiga toxin-producing Escherichia coli during the slaughter of cattle using molecular typing combined with epidemiologic data.

Investigation on the distribution and biological characteristics of Shiga-toxin producing Escherichia coli (STEC) during beef processing is essential for in-plant critical control points and food safety risk assessment. Serogroups and subtypes of stx genes of STEC strains isolated from beef processing lines were first investigated. Identification to cross-contamination among different sampling sites was further conducted by combining multilocus sequence typing (MLST) with the previous distribution and characterization data. The PCR-positive rate for STEC in 435 samples from two slaughter plants in China was 14.3% and the isolation rate for the 62 PCR positive and the entire set of 435 samples were 26% and 3.68% respectively. The existence of serotype O157:H7 (33%) and serogroups O121 (42%) and O26 (21%) as well as the high detection rate of high pathogenic gene stx2a (68%) in these serogroups indicated potential risk to the safety of beef. Traceability analysis showed that hide plays a critical role in cross-contamination between feces, lairage pens and post-washing carcasses from a molecular perspective. Intervening measures revolves around de-hiding should be involved in the in-plant safety control policy according to the tracing analysis.

Effects of different protein phosphorylation levels on the tenderness of different ultimate pH beef.

This study investigated the relationship between tenderness and protein phosphorylation levels of normal ultimate pH (pHu, 5.4-5.8, NpHu), intermediate pHu (5.8-6.2, IpHu) and high pHu (≥6.2, HpHu) Longissimus lumborum from beef. During 21 d of ageing, the HpHu group had the lowest Warner-Bratzler shear force (WBSF) values, while the IpHu group showed the highest and even after 21 days of ageing still had high levels. In the late stage of the 24 h post-mortem period the faster degradation rate of troponin T and earlier activation of caspase 9 in the HpHu group were the key reasons for the lower WBSF compared with the NpHu and IpHu groups. The activity of caspase 3 cannot explain the tenderness differences between IpHu and HpHu groups, since their activities did not show any difference. At 24 h post-mortem, 17 common differential phosphorylated peptides were detected among pHu groups, of which nine were associated with pHu and WBSF. The higher phosphorylation level of glycogen synthase may have caused the delay of meat tenderization in the IpHu group.

Sous vide cooking improved the physicochemical parameters of hot-boned bovine semimembranosus muscles.

The physicochemical parameters of hot-boned bovine semimembranosus muscles after sous vide cooking were investigated. Hot-boned or wet-aged steaks were collected, and cooked by different cooking methods, including sous vide (57 °C, 11 h, SV), grilling (at 200 °C to the central temperature of 72 °C, GR) or boiling (100 °C, 2 h, BO). The meat color, tenderness, water-holding capacity, degree of oxidation, myoglobin denaturation and sensory quality traits were determined, as well as the changes in the microstructure. Compared to other cooking methods, SV reduced the degree of oxidation and muscle shortening, and significantly improved the water holding capacity (WHC), tenderness, connective tissue content and overall acceptability for both hot-boned and wet-aged steaks. The oxidation and muscle shortening were reduced in hot-boned SV steaks (P < 0.05), and the water-holding capacity and sensory scores for juiciness, connective tissue content and overall acceptability were increased (P < 0.05) compared to the wet-aged steaks. The combination of hot-boning and SV cooking resulted in an acceptable tenderness, better overall sensory acceptability and higher WHC than other combinations of muscle states and cooking methods. Therefore, it is a good choice to cook hot-boned semimembranosus muscles using SV to improve the eating quality, which can eliminate the need for aging, benefiting the beef industry.

Effect of resveratrol on skeletal slow-twitch muscle fiber expression via AMPK/PGC-1α signaling pathway in bovine myotubes.

The purpose of this study was to evaluate the impact of resveratrol on slow-twitch muscle fiber expression in bovine myotubes. The results revealed that resveratrol enhanced slow myosin heavy chain (MyHC) and suppressed fast MyHC protein expression, accompanied by increased MyHC I/IIa and decreased MyHC IIx/IIb mRNA levels in bovine myotubes (P < 0.05). Resveratrol also enhanced the activities of succinic dehydrogenase (SDH), malate dehydrogenase (MDH) and the mitochondrial DNA (mtDNA) content, but reduced lactate dehydrogenase (LDH) activity (P < 0.05). Meanwhile, the protein and gene expression of AMPK, SIRT1 and PGC-1α were upregulated by resveratrol (P < 0.05). Furthermore, PGC-1α inhibitor SR-18292 could attenuate resveratrol-induced muscle fiber conversion from fast-twitch to slow-twitch. These results suggest that resveratrol might promote muscle fiber type transition from fast-twitch to slow-twitch through the AMPK/PGC-1α signaling pathway and mitochondrial biogenesis in bovine myotubes.

Potential inhibitory effect of carbon dioxide on the spoilage behaviors of Pseudomonas fragi in high-oxygen packaged beef during refrigerated storage.

Pseudomonas fragi is a dominant meat spoilage organism under high-oxygen modified atmosphere packaging (HiOx-MAP). This work investigated the effects of CO2 on P. fragi growth and the related spoilage phenomena of HiOx-MAP beef. Minced beef incubated with P. fragi T1, a strain owning the strongest spoilage potential among isolates, was stored under CO2-enriched HiOx-MAP (TMAP; 50% O2/40% CO2/10% N2) or non-CO2 HiOx-MAP (CMAP; 50% O2/50% N2) at 4 °C for 14 days. Compared to CMAP, TMAP maintained sufficient O2 levels to endow beef with higher a* values and meat color stability due to lower P. fragi counts from day 1 (P < 0.05). TMAP samples also showed lower (P < 0.05) lipase activity and protease activity within 14-days and 6-days than CMAP samples respectively. TMAP delayed the significantly increased pH and total volatile basic nitrogen contents occurred in CMAP beef during storage. Despite TMAP markedly promoted the lipid oxidation associated with higher concentrations of hexanal and 2,3-octanedione than CMAP (P < 0.05), TMAP beef retained an acceptable organoleptic odor due to a CO2-inhibition on the microbial-induced 2,3-butanedione and ethyl 2-butenoate formation. This study provided a comprehensive insight into the antibacterial mechanism of CO2 on P. fragi in HiOx-MAP beef.

Preliminary investigation into the prediction of indicators of beef spoilage using Raman and Fourier transform infrared spectroscopy.

The objective of this study was to assess the potential to predict the microbial beef spoilage indicators by Raman and Fourier transform infrared (FT-IR) spectroscopies. Vacuum skin packaged (VSP) beef steaks were stored at 0 °C, 4 °C, 8 °C and under a dynamic temperature condition (0 °C âˆ¼ 4 °C âˆ¼ 8 °C, for 36 d). Total viable count (TVC) and total volatile basic nitrogen (TVB-N) were obtained during the storage period along with spectroscopic data. The Raman and FTIR spectra were baseline corrected, pre-processed using Savitzky-Golay smoothing and normalized. Subsequently partial least squares regression (PLSR) models of TVC and TVB-N were developed and evaluated. The root mean squared error (RMSE) ranged from 0.81 to1.59 (log CFU/g or mg/100 g) and the determination coefficient (R2) from 0.54 to 0.75. The performance of PLSR model based on data fusion (combination of Raman and FT-IR data) is better than that based on Raman spectra and similar to that of FT-IR. Overall, Raman spectroscopy, FT-IR spectroscopy, and a combination of both exhibited a potential for the prediction of the beef spoilage.

The biological effect of a beef-derived Latilactobacillus sakei on beef steaks during chilled storage.

The aim of this study was to investigate the biological inhibiting effect of a beef-derived Latilactobacillus sakei (RS-25) on the spoilage of beef steaks in overwrapped packaging during the 12 days of storage at 4°C. Beef quality as well as microbial indicators were determined at different intervals during the storage after the inoculation of RS-25 at the 6 log CFU/g, and the high-throughput sequencing was applied to investigate the changes of microbial community structure during the storage. The inoculation of RS-25 on beef had no effect (p > .05) on pH, TBARS, and TVB-N during storage indicating the weak effect of such strain on the eat quality. Furthermore, the rise of L* and the delayed decline of a* and b* reveal the protection effect of RS-25 on the meat color. RS-25 reduced the re-contaminated Salmonella typhimurium by 1.16 log CFU/g (p < .01), and the growth of Brochothrix thermosphacta was also inhibited but no inhibition was found on the Pseudomonas spp. at the first 6 days of storage. The inhibiting effect of RS-25 was covered by the rapid growth of other microorganism during the following 6 days of storage. Consistent with the microbial counts results, high-throughput sequencing analysis confirmed that the inoculated L. sakei RS-25 was dominant at first 6 days, and then replaced by Pseudomonas spp. The findings obtained from the current study may provide basic information for the further application of bioprotective bacteria in preservation of beef steaks in the overwrapped packaging.

Mechanisms and applications of probiotics in prevention and treatment of swine diseases.

Probiotics can improve animal health by regulating intestinal flora balance, improving the structure of the intestinal mucosa, and enhancing intestinal barrier function. At present, the use of probiotics has been a research hotspot in prevention and treatment of different diseases at home and abroad. This review has summarized the researchers and applications of probiotics in prevention and treatment of swine diseases, and elaborated the relevant mechanisms of probiotics, which aims to provide a reference for probiotics better applications to the prevention and treatment of swine diseases.

High-pressure processing and modified atmosphere packaging combinations for the improvement of dark, firm, and dry beef quality and shelf-life.

This study investigated the effects of high-pressure processing (HPP) and modified atmosphere packaging (MAP) on 'dark, firm, and dry' (DFD) beef. To optimize the HPP, beef steaks (n = 180) were first processed at different pressures (0.1, 200, 300, 400, 500 MPa). It was found that 400 MPa enhanced DFD beef color and shelf-life. This optimized HPP (400 MPa) was combined with 3 MAP formulations, in a second study (40, 60, or 80% O2-MAP), to determine their effect on DFD beef steaks. HPP (400 MPa) combined with MAP improved DFD beef L* and a*, color scores, and delayed discoloration (P < 0.01). Total plate counts for DFD beef held under 60% O2-MAP was ≤6 log10 CFU/g, even after 14 d of chilled storage. These same samples had shear force and TBARS values significantly lower than observed for DFD beef held under 80% O2-MAP. HPP (400 MPa) combined with 60% O2-MAP is recommended to improve DFD beef quality and shelf-life.

Exploratory study on the potential regulating role of Peroxiredoxin 6 on proteolysis and relationships with desmin early postmortem.

The objective of the present study was to explore the effect of Peroxiredoxin 6 (Prdx 6) on beef tenderization during the early postmortem period. The longissimus lumborum (LL) were obtained at 45 min postmortem from 6 beef carcasses and then incubated with or without the inhibitor of Prdx6 (NSC348884) for different times, followed by incubation with or without the H2O2 (simulation of oxidative stress). The expression of Prdx6, proteolysis indicated by desmin degradation, cell apoptosis rate and expression of caspases were measured. The results indicated that the inhibitor significantly reduced the Prdx6 level, while the cells adaptively increased Prdx6 expression to resist the oxidative stress caused by H2O2. Moreover, the samples in which Prdx6 was inhibited demonstrated more severe desmin degradation accompanied by a higher apoptosis rate which was induced by the increase in caspase degradation as well as the ratio of Bax/Bcl-2. These results demonstrated that inhibiting Prdx6 could promote cell apoptosis and further accelerate beef tenderization.

Influence of low-energy electron beam irradiation on the quality and shelf-life of vacuum-packaged pork stored under chilled and superchilled conditions.

This study aimed to explore the effects of different levels (0, 8, and 12 kGy) of low-energy electron beam (LEEB; 0.2 MeV) on the quality and shelf-life of vacuum-packaged pork stored under chilled and superchilled conditions for 30 days. LEEB irradiation promoted lipid and protein oxidation, regardless of the levels used. Combined 8 kGy LEEB irradiation with superchilled storage significantly elevated a* values and preserved acceptable sensory characteristics of pork until day 30, unlike the other treatment combinations. This synergistic treatment also significantly reduced total viable counts and total volatile basic nitrogen contents, and thus extended the shelf-life of vacuum-packaged pork to >30 days, which may be due to the antibacterial effects on Photobacterium, Carnobacterium and Lactobacillus. In conclusion, it is recommended that the 8 kGy LEEB irradiation level be applied to pork and that it should be used in combination with superchilled storage to preserve meat quality and extend the shelf-life of vacuum-packaged pork.

Effect of dietary resveratrol supplementation on muscle fiber types and meat quality in beef cattle.

In order to investigate the effect of dietary resveratrol supplementation on muscle fiber types and meat quality in beef cattle, a feeding experiment was undertaken. Longissimus lumborum, Psoas major and Semitendinosus muscles were collected 24 h post-mortem from two groups of cattle, which were fed with a total mixed ration (Control - CON) or supplemented with resveratrol (5 g/animal/day, RES) for 120 d before slaughter. The results showed that dietary resveratrol increased the gene expression of MyHC I and enhanced the proportion of type I fibers in three muscles. The cooking loss and Warner-Bratzler shear force of all muscles during aging for 21 days were decreased. However, the increased proportion of type I fibers resulted in a darker initial color, but did improve color stability, as the a* value of RES samples was lower initially but higher in the later stage of aging. This study indicates the supplementation potential of resveratrol for beef cattle for tenderness and color stability.

Processing interventions for enhanced microbiological safety of beef carcasses and beef products: A review.

Chilled beef is inevitably contaminated with microorganisms, starting from the very beginning of the slaughter line. A lot of studies have aimed to improve meat safety and extend the shelf life of chilled beef, of which some have focused on improving the decontamination effects using traditional decontamination interventions, and others have investigated newer technologies and methods, that offer greater energy efficiency, lower environmental impacts, and better assurances for the decontamination of beef carcasses and cuts. To inform industry, there is an urgent need to review these interventions, analyze the merits and demerits of each technology, and provide insight into 'best practice' to preserve microbial safety and beef quality. In this review, the strategies and procedures used to inhibit the growth of microorganisms on beef, from slaughter to storage, have been critiqued. Critical aspects, where there is a lack of data, have been highlighted to help guide future research. It is also acknowledge that different intervention programs for microbiological safety have different applications, dependent on the initial microbial load, the type of infrastructures, and different stages of beef processing.

The underlying mechanisms of the effect of superchilling on the tenderness of beef Longissimus lumborum.

This study investigated the effect of superchilling (-30 °C until the core temperature achieved -3 °C, then stored at -1 °C until 24 h, SC) on the tenderness of hot boned beef M. longissimus lumborum (LL), with very fast chilling (-30 °C until the core temperature achieved 0 °C, then stored at -1 °C until 24 h, VFC) and conventional chilling (0- 4 °C for 24 h, CC) as the controls. The lowest initial shear force values were obtained in SC samples compared to those from the VFC and CC treatments (P < 0.05). Clear freezing damage of muscle fibers and more myofibril fragmentation were found in SC samples compared with the other samples early post-mortem. Moreover, SC samples showed the highest level of inosine 5-monophosphate at 3 h post-mortem (P < 0.05). A reduced glycolysis rate (as evidenced by lactate content) was also found in SC treated samples suggesting little contribution of glycolysis on the tenderization of SC.