Bactericidal effect of ultrasound on glutinous rice during soaking and its influence on physicochemical properties of starch and quality characteristics of sweet dumplings.

The soaking process of glutinous rice allows the growth and reproduction of microorganisms, which can easily cause food safety problems. In this work, the effects of different ultrasonic powers (150 W, 300 W, 450 W, and 600 W) on the bactericidal effect of glutinous rice, the physicochemical properties of starch and the quality characteristics of sweet dumplings were studied. Compared with soaking for 0 and 2 h, sonication of glutinous rice after soaking for 4 h was more effective at reducing the number of microorganisms in soaked glutinous rice, and the bactericidal effect increased with increasing ultrasound intensity. After 30 min, the total number of bacteria decreased by 2.04 log CFU/g. Moreover, ultrasonic treatment destroys the grain structure of glutinous rice starch, resulting in the formation of dents and cracks on the starch surface, increasing the amylose content, improving its expansion, reducing its short-range order and relative crystallinity, and altering its gelatinization characteristics. In addition, ultrasonic treatment increased the soup transparency of sweet dumplings from 51.8 % to 63.95 %, reducing their hardness, chewiness and adhesiveness. In summary, ultrasonic treatment can not only effectively kill microorganisms in soaked glutinous rice but also improve the quality of glutinous rice dumplings by changing the physicochemical properties of glutinous rice starch. The results of this study provide theoretical support for the application of ultrasonic technology in glutinous rice food production.

Co-culture fermentation by Saccharomycopsis fibuligera and lactic acid bacteria improves bioactivity and aroma profile of wheat bran and the bran-containing Chinese steamed bread.

Co-culture fermentation with yeast and lactic acid bacteria (LAB) exhibits advantages in improving the bioactivity and flavor of wheat bran compared to single-culture fermentation, showing application potentials in bran-containing Chinese steamed bread (CSB). To explore the effects of combination of yeast and different LAB on the bioactivity and flavor of fermented wheat bran, this study analyzed the physicochemical properties, phytate degradation capacity, antioxidant activities, and aroma profile of wheat bran treated with co-culture fermentation by Saccharomycopsis fibuligera and eight different species of LAB. Further, the phenolic acid composition, antioxidant activities, texture properties, aroma profile, and sensory quality of CSB containing fermented wheat bran were evaluated. The results revealed that co-culture fermentation brought about three types of volatile characteristics for wheat bran, including ester-feature, alcohol and acid-feature, and phenol-feature, and the representative strain combinations for these characteristics were S. fibuligera with Limosilactobacillus fermentum, Pediococcus pentosaceus, and Latilactobacillus curvatus, respectively. Co-culture fermentation by S. fibuligera and L. fermentum for 36 h promoted acidification with a phytate degradation rate reaching 51.70 %, and improved the production of volatile ethyl esters with a relative content of 58.47 % in wheat bran. Wheat bran treated with co-culture fermentation by S. fibuligera and L. curvatus for 36 h had high relative content of 4-ethylguaiacol at 52.81 %, and exhibited strong antioxidant activities, with ABTS•+ and DPPH• scavenging rates at 65.87 % and 69.41 %, respectively, and ferric reducing antioxidant power (FRAP) at 37.91 µmol/g. In addition, CSB containing wheat bran treated with co-culture fermentation by S. fibuligera and L. fermentum showed a large specific volume, soft texture, and pleasant aroma, and received high sensory scores. CSB containing wheat bran treated with co-culture fermentation by S. fibuligera and L. curvatus, with high contents of 4-ethylguaiacol, 4-vinylguaiacol, ferulic acid, vanillin, syringaldehyde, and protocatechualdehyde, demonstrated strong antioxidant activities. This study is beneficial to the comprehensive utilization of wheat bran resources and provides novel insights into the enhancement of functions and quality for CSB.

Comparative genomic analysis and multilocus sequence typing of Staphylococcus aureus reveals candidate genes for low-temperature tolerance.

Staphylococcus aureus is one of the most frequently detected foodborne pathogens in cold chain foods. Worryingly, small colony variants (SCVs) can survive in cold environments for a long time and can revert to rapidly growing cells in suitable environments, causing serious food safety issues. This study investigated the underlying mechanism of SCV formation at low temperature (4 °C) via comparative genomics. Multilocus sequence typing (MLST) of 105 strains of S. aureus was divided into 9 sequence types. The ST352 strains exhibited the greatest tolerance to low temperature, with a mean reduction in survival rate of 10.34 % (p < 0.05). Comparative genomics revealed a total of 1941 core genes in the three S. aureus strains, and BB-1 had 468 specific genes, which were enriched mainly in translation, DNA recombination, DNA repair, metabolic pathways, two-component systems, and quorum sensing. Molecular docking analysis revealed that the binding of the RsbW protein to the SigB protein of BB-1 decreased due to base mutations in rsbW, while the binding to the RsbV protein was enhanced. In addition, the results of real-time quantitative PCR showed that the RsbV-RsbW/SigB system of BB-1 may play a role in the low-temperature survival of S. aureus and the formation of SCVs. These results suggest that genes specific to BB-1 may contribute to the mechanism of adaptation to low temperature and the formation of SCVs. This study helps elucidate the causes of SCV formation by S. aureus at low temperature at the molecular level and provides a basis for exploring the safety control of cold chain food environments.

Antibiotic resistance profile of Helicobacter pylori to 14 antibiotics: a multicenter study in Fujian, China.

Background and aim: Efficacy of Helicobacter pylori (H. pylori) eradication is related to the local antimicrobial resistance epidemiology. We aimed to investigate the antibiotic resistance of H. pylori in Fujian, China. Methods: H. pylori-infected patients in four centers were enrolled in the study from Oct 2019 to Jan 2022. The bacteria were isolated, cultured and identified from the biopsy of patients' gastric mucosa samples. Antimicrobial susceptibility testing was performed by a modified broth microdilution method for H. pylori to seven guideline-recommended antibiotics and seven potential choices for H. pylori eradication. Results: A total of 205 H. pylori strains were isolated. The resistance rates of amoxicillin (AMX), amoxicillin and clavulanate potassium (AMC), cefixime (CFM), gentamicin (GEN), tetracycline (TET), doxycycline (DOX), azithromycin (AZM), clarithromycin (CLR), levofloxacin (LVFX), sparfloxacin (SPFX), metronidazole (MTZ), tinidazole (TID), rifampicin (RFP) and furazolidone (FZD) were 11.22%, 12.20%, 7.32%, 12.20%, 4.88%, 4.39%, 44.39%, 43.90%, 30.24%, 21.46%, 40.98%, 45.85%, 5.37% and 10.24%, respectively. The rates of pan-sensitivity, single, double, triple and multiple resistance for seven guideline-recommended antibiotics were 32.68%, 30.24%, 13.17%, 7.76%, and 14.15%, respectively. The main double-resistance patterns were CLR+MTZ (10/205, 5%) and CLR+LVFX (9/205, 4%). The main triple-resistance pattern was CLR+MTZ+ LVFX (15/205, 7%). Conclusions: In Fujian, the prevalence of H. pylori resistance to AZM, CLR, LVFX, SPFX, MTZ, and TID was high, whereas that to AMX, AMC, GEN, CFM, TET, DOX, RFP and FZD was relatively low. CFM and DOX are promising new choices for H. pylori eradication.

Mechanism underlying the weakening effect of ß-glucan on the gluten system.

The high ß-glucan content in barley disrupts the gluten network in dough. Scanning electron microscopy (SEM), differential scanning calorimetry (DSC), fourier transform infrared spectroscopy (FTIR), and solid-state nuclear magnetic resonance (NMR) techniques were used to clarify how ß-glucan affected the quality of the gluten network structure with ß-glucan contents of 0-2%. The results suggest that the physical hindrance of the ß-glucan gel destroyed the formation of the gluten network structure. When 1.0-2.0% ß-glucan was added, the percentage of α-helical structures increased significantly. When the added amount of ß-glucan reached 2.0%, the sulfhydryl group (SH) content increased from 8.06 to 10.27 µmol/g, and the disulfide bond (SS) content decreased from 240.09 to 217.38 µmol/g. The interaction between ß-glucan and gluten mainly resulted from the interaction of electron-withdrawing groups, such as carbonyl groups (CO) and double bond carbons (CC), and carbon atoms on the side chains of ß-glucan, which play an important role in the central structure of glutenin.

Transcriptome analysis reveals the molecular mechanism of cinnamaldehyde against Bacillus cereus spores in ready-to-eat beef.

The purpose of this study was to investigate the antibacterial effect and mechanism of cinnamaldehyde on Bacillus cereus spores in ready-to-eat beef. The colour difference and texture of the ready-to-eat beef supplemented with cinnamaldehyde did not differ greatly from the colour and texture of the blank beef. However, cinnamaldehyde has an effective antibacterial effect on the total number of bacterial colonies and B. cereus spores in ready-to-eat beef. Transmission electron microscopy (TEM) analysis revealed that the cell membrane of B. cereus was disrupted by cinnamaldehyde, leading to leakage of intracellular components. Transcriptome sequencing (RNA-seq) indicated that the B. cereus spore resistance regulation system (sigB, sigW, rsbW, rsbV, yfkM and yflT) and phosphoenolpyruvate phosphotransferase system (PTS) (ptsH, ptsI and ptsG) respond positively to cinnamaldehyde in an adverse environment. Intracellular disorders due to damage to the cell membrane involve some transporters (copA, opuBA and opuD) and some oxidative stress systems (ywrO, scdA and katE) in the regulation of the body. However, downregulation of K+ transport channels (kdpD and kdpB), osmotic pressure regulation (opuE) and some oxidative stress (norR and srrA)-related genes may accelerate spore apoptosis. In addition, cinnamaldehyde also effectively inhibits the spore germination-related genes (smc, mreB and gerE). This study provides new insights into the molecular mechanism of the antibacterial effect of cinnamaldehyde on B. cereus spores in ready-to-eat beef.

Comparative Transcriptomic Analysis of Staphylococcus aureus Reveals the Genes Involved in Survival at Low Temperature.

In food processing, the temperature is usually reduced to limit bacterial reproduction and maintain food safety. However, Staphylococcus aureus can adapt to low temperatures by controlling gene expression and protein activity, although its survival strategies normally vary between different strains. The present study investigated the molecular mechanisms of S. aureus with different survival strategies in response to low temperatures (4 °C). The survival curve showed that strain BA-26 was inactivated by 6.0 logCFU/mL after 4 weeks of low-temperature treatment, while strain BB-11 only decreased by 1.8 logCFU/mL. Intracellular nucleic acid leakage, transmission electron microscopy, and confocal laser scanning microscopy analyses revealed better cell membrane integrity of strain BB-11 than that of strain BA-26 after low-temperature treatment. Regarding oxidative stress, the superoxide dismutase activity and the reduced glutathione content in BB-11 were higher than those in BA-26; thus, BB-11 contained less malondialdehyde than BA-26. RNA-seq showed a significantly upregulated expression of the fatty acid biosynthesis in membrane gene (fabG) in BB-11 compared with BA-26 because of the damaged cell membrane. Then, catalase (katA), reduced glutathione (grxC), and peroxidase (ahpC) were found to be significantly upregulated in BB-11, leading to an increase in the oxidative stress response, but BA-26-related genes were downregulated. NADH dehydrogenase (nadE) and α-glucosidase (malA) were upregulated in the cold-tolerant strain BB-11 but were downregulated in the cold-sensitive strain BA-26, suggesting that energy metabolism might play a role in S. aureus under low-temperature stress. Furthermore, defense mechanisms, such as those involving asp23, greA, and yafY, played a pivotal role in the response of BB-11 to stress. The study provided a new perspective for understanding the survival mechanism of S. aureus at low temperatures.

Minimum inhibitory concentrations of commonly used antibiotics against Helicobacter Pylori: A multicenter study in South China.

AIM: To determine the minimum inhibitory concentrations (MICs) of commonly used antibiotics against Helicobacter Pylori (H. pylori) in South China and compare their resistance rates by using EUCAST breakpoints and other breakpoints. METHODS: Patients who had not previously received H. pylori treatment in clinical centers in South China were enrolled in this study from 2017 to 2020. Gastric biopsies were obtained for H. pylori culture. The MICs of amoxicillin (AMX), clarithromycin (CLA), metronidazole (MTZ), levofloxacin (LEV), tetracycline (TET) and furazolidone (FZD) were tested by broth microdilution method and assessed by two different breakpoints. ATCC43504 standard strain served as a control. RESULTS: A total of 208 H. pylori strains were isolated from patients' biopsy samples. The MICs of AMX, CLA, MTZ, LEV, TET and FZD for H. pylori were 0.0156-256mg/L (MIC50 0.125mg/L, MIC90 4mg/L), 0.0156- >256 mg/L (MIC50 0.0312mg/L, MIC90 64mg/L), 0.0156- >256mg/L (MIC50 8mg/L, MIC90 256mg/L), 0.0156-256mg/L (MIC50 0.25mg/L, MIC90 16mg/L), 0.0156-256mg/L (MIC50 0.0625mg/L, MIC90 4mg/L), and 0.0156- >256mg/L (MIC50 0.0312mg/L, MIC90 2mg/L), respectively. The MICs of AMX, CLA, MTZ, LEV, TET and FZD for ATCC43504 strain were 0.25mg/L, 0.0625mg/L, 64mg/L, 0.5mg/L, 1mg/L and 0.25mg/L, respectively. The resistance rate of FZD was 11.05%. The overall resistance rates according to EUCAST breakpoints and other breakpoints were 57.21% and 14.90% for AMX (p<0.001), 38.94% and 38.94% for CLA (p = 1), 39.42% and 50.96% for MTZ (p<0.001), 12.98% and 10.58% for TET (p = 0.025), 35.10% and 35.10% for LEV (p = 1), respectively. CONCLUSIONS: Our results demonstrate that AMX, FZD, and TET, but not MTZ, CLR or LEV, showed good anti-H. pylori activity in vitro in South China. When different breakpoints were used, similar results were found with CLA, and LEV, but not with AMX, MTZ, or TET.

Diversity of bacterial communities in traditional sourdough derived from three terrain conditions (mountain, plain and basin) in Henan Province, China.

To elucidate the bacterial community composition of sourdoughs from different terrain conditions, thirty-two Chinese traditional sourdough samples were collected from three terrain conditions (mountain, plain and basin) in Henan Province. High-throughput sequencing and culture-dependent approaches were employed to identify the bacterial diversity of the sourdough samples. A total of two hundred and six isolates were characterized via 16S rRNA gene sequencing. Pediococcus pentosaceus was isolated from every sample and was the predominant species in the sourdough samples, accounting for 58% of the relative abundance. High-throughput sequencing revealed that the predominant genera (mainly Pediococcus) in the basin group were significantly different from those in the mountain and plain groups. The genus Lactobacillus was predominant in the plain and mountain sourdough samples. Pediococcus pentosaceus was the absolute dominant strain in the basin sourdough samples. Acetobacter, which was widely distributed only in mountain samples, was recognized as the representative genus of the mountain samples. Moreover, we first reported Gluconobacter oxydans in sourdough. This study provided insight into the bacterial diversity of sourdough from three terrain conditions (mountain, plain and basin) in Henan Province and could serve as a reference for the isolation of desired bacterial strains.

The iTRAQ-based chloroplast proteomic analysis of Triticum aestivum L. leaves subjected to drought stress and 5-aminolevulinic acid alleviation reveals several proteins involved in the protection of photosynthesis.

BACKGROUNDS: The perturbance of chloroplast proteins is a major cause of photosynthesis inhibition under drought stress. The exogenous application of 5-aminolevulinic acid (ALA) mitigates the damage caused by drought stress, protecting plant growth and development, but the regulatory mechanism behind this process remains obscure. RESULTS: Wheat seedlings were drought treated, and the iTRAQ-based proteomic approach was employed to assess the difference in chloroplast protein content caused by exogenous ALA. A total of 9499 peptides, which could be classified into 2442 protein groups, were identified with ≤0.01 FDR. Moreover, the contents of 87 chloroplast proteins was changed by drought stress alone compared to that of the drought-free control, while the contents of 469 was changed by exogenous ALA application under drought stress compared to that of drought stress alone. The Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis results suggested that the ALA pretreatment adjusted some biological pathways, such as metabolic pathways and pathways involved in photosynthesis and ribosomes, to enhance the drought resistance of chloroplasts. Furthermore, the drought-promoted H2O2 accumulation and O2- production in chloroplasts were alleviated by the exogenous pretreatment of ALA, while peroxidase (POD) and glutathione peroxidase (GPX) activities were upregulated, which agreed with the chloroplast proteomic data. We suggested that ALA promoted reactive oxygen species (ROS) scavenging in chloroplasts by regulating enzymatic processes. CONCLUSIONS: Our results from chloroplast proteomics extend the understanding of the mechanisms employed by exogenous ALA to defend against drought stress in wheat.

Synergistic effect of cinnamaldehyde on the thermal inactivation of Listeria monocytogenes in ground pork.

The aim of this study was to statistically evaluate the effect of a naturally food-derived cinnamaldehyde on the thermal inactivation of Listeria monocytogenes in ground pork. This study combined four concentrations of cinnamaldehyde (0, 0.1, 0.5, and 1.0% vol/wt) and four temperatures (55, 60, 65, and 70 ℃) to predict the thermal inactivation curves of L. monocytogenes. The Weibull model successfully described the primary thermal inactivation using the Integrated Pathogen Modeling Program. These results statistically proposed that the cinnamaldehyde supplementation in ground pork attenuates the thermo-tolerance of L. monocytogenes. The time for achieving a 5-log10 reduction of L. monocytogenes declined from 28.14 to 17.35 min at 55 ℃ when the ground pork sample was supplemented by 1% cinnamaldehyde, while the time declined from 1.95 to 0.34 min at 70 ℃. Thereafter, based on the 5.0-log10 lethality, secondary models were fitted by a selected polynomial model. The transmission electron microscopy revealed that cinnamaldehyde causes serious damage to membrane integrity and increases the occurrence of cell membrane rupture and leakage of cytoplasmic content under thermal treatment. Our model represents a mathematical tool that will help meat-product manufacturers to improve the efficacy of thermal processing ground pork supplemented with cinnamaldehyde.

Effect of potassium alum addition on the quality of potato starch noodles.

The objective of this study was to investigate the effects and mechanism of potassium alum (PA) on potato starch noodles to provide a theoretical basis for future searches for new alternatives for PA that are not harmful to the body. The amount of amylose leakage significantly decreased with the PA addition. SEM showed that the honeycomb structure of potato starch noodles with 0.1% PA was diminished. The number of internal pores and average area decreased from 92 and 580.09 units to 72 and 652.58 units, respectively. NMR indicated that the water holding capacity of potato starch noodles containing PA was greater than that of the control. XRD showed that the crystal structure of potato starch noodles was destroyed and that the crystallinity of potato starch noodles with PA increased from 16.65 to 19.64%. G' and G'' increased, but tanδ decreased. FT-IR showed that PA caused the O-H stretching vibration in the absorption peak to move in the high-field direction and that the ionic bond was dominant in the PA-potato starch interaction system.

Application of Lactobacillus as Adjunct Cultures in Wheat Dough Fermentation.

The purpose of this study was to evaluate the potential use of Lactobacillus from traditional Chinese sourdoughs of different regions as original adjunct cultures in the steamed bread making process. The effects of Lactobacillus on dough and steamed bread were evaluated. Some differences were obtained in the parameters of fermented dough (organic acid content, rheofermentative parameters, pH, and total titratable acidity [TTA]) and steamed bread (hardness, specific volume, organic acid content, shape, color, pH, TTA and sensory score) made with five different Lactobacillus strains (Pediococcus pentosaceus, Lactobacillus fermentum, Weissella confusa, Lactobacillus crustorum, and Pediococcus acidilactici). Steamed bread made with P. pentosaceus showed a significant increase in specific volume (from 2.19 to 2.41) and a decrease in hardness (from 3,158 g to 2,301 g). Dough leavened by L. fermentum showed significantly higher amounts of lactic acid and acetic acid than dough inoculated with W. confusa, which had values similar to those of the control. The dough fermented by P. pentosaceus exhibited the most gas production (1,811 mL), which is an important index of streamed bread quality. Our research provides a reference for making steamed bread with Lactobacillus. PRACTICAL APPLICATION: Probiotic Lactobacillus is widely used to produce fermenting foods and has been used in bread processing to improve the quality and characteristics of bread. Steamed bread is one of the most popular fermented foods in northern China. However, there is little information about the application of Lactobacillus in Chinese steamed bread. This study explored the potential use of Lactobacillus to improve the characteristics and quality of steamed bread.

Comparative Proteomic and Morphological Change Analyses of Staphylococcus aureus During Resuscitation From Prolonged Freezing.

When frozen, Staphylococcus aureus survives in a sublethally injured state. However, S. aureus can recover at a suitable temperature, which poses a threat to food safety. To elucidate the resuscitation mechanism of freezing survived S. aureus, we used cells stored at -18°C for 90 days as controls. After resuscitating the survived cells at 37°C, the viable cell numbers were determined on tryptic soy agar with 0.6% yeast extract (TSAYE), and the non-injured-cell numbers were determined on TSAYE supplemented with 10% NaCl. The results showed that the total viable cell number did not increase within the first 3 h of resuscitation, but the osmotic regulation ability of freezing survived cells gradually recovered to the level of healthy cells, which was evidenced by the lack of difference between the two samples seen by differential cell enumeration. Scanning electron microscopy (SEM) showed that, compared to late exponential stage cells, some frozen survived cells underwent splitting and cell lysis due to deep distortion and membrane rupture. Transmission electron microscopy (TEM) showed that, in most of the frozen survived cells, the nucleoids (low electronic density area) were loose, and the cytoplasmic matrices (high electronic density area) were sparse. Additionally, a gap was seen to form between the cytoplasmic membranes and the cell walls in the frozen survived cells. The morphological changes were restored when the survived cells were resuscitated at 37°C. We also analyzed the differential proteome after resuscitation using non-labeled high-performance liquid chromatography-mass spectrometry (HPLC-MS). The results showed that, compared with freezing survived S. aureus cells, the cells resuscitated for 1 h had 45 upregulated and 73 downregulated proteins. The differentially expressed proteins were functionally categorized by gene ontology enrichment, KEGG pathway, and STRING analyses. Cell membrane synthesis-related proteins, oxidative stress resistance-related proteins, metabolism-related proteins, and virulence factors exhibited distinct expression patterns during resuscitation. These findings have implications in the understanding of the resuscitation mechanism of freezing survived S. aureus, which may facilitate the development of novel technologies for improved detection and control of foodborne pathogens in frozen food.

Effects of ZnO nanoparticle-coated packaging film on pork meat quality during cold storage.

BACKGROUND: There has been limited research on the use of ZnO nanoparticle-coated film for the quality preservation of pork meat under low temperature. In the present study, ZnO nanoparticles were mixed with sodium carboxymethyl cellulose (CMC-Na) to form a nanocomposite film, to investigate the effect of ZnO nanoparticle-coated film on pork meat quality and the growth of bacteria during storage under low temperature. RESULTS: When ZnO nanoparticle-coated film was used as the packaging material for pork meat for 14 days of cold storage at 4 °C, the results demonstrated a significant effect on restricting the increases in total volatile basic nitrogen and pH levels, limiting the decreases of lightness (increased L* value) and redness (increased a* value), and maintaining the water-holding capacity compared to the control pork samples (P < 0.05). The present study also discovered that the ZnO nanoparticle-coated film restrained the increase in total plate count (TPC). When Staphylococcus aureus was used as the representative strain, scanning electron microscopy revealed that ZnO nanoparticles increased the occurrence of cell membrane rupture under cold conditions. CONCLUSION: ZnO nanoparticle-coated film helps retain the quality of pork meat during cold storage by increasing the occurrence of microorganism injury. © 2016 Society of Chemical Industry.

Identification of Circular RNAs and Their Targets in Leaves of Triticum aestivum L. under Dehydration Stress.

Circular RNAs (circRNAs) are a type of newly identified non-coding RNAs through high-throughput deep sequencing, which play important roles in miRNA function and transcriptional controlling in human, animals, and plants. To date, there is no report in wheat seedlings regarding the circRNAs identification and roles in the dehydration stress response. In present study, the total RNA was extracted from leaves of wheat seedlings under dehydration-stressed and well-watered conditions, respectively. Then, the circRNAs enriched library based deep sequencing was performed and the circRNAs were identified using bioinformatics tools. Around 88 circRNAs candidates were isolated in wheat seedlings leaves while 62 were differentially expressed in dehydration-stressed seedlings compared to well-watered control. Among the dehydration responsive circRNAs, six were found to act as 26 corresponding miRNAs sponges in wheat. Sixteen circRNAs including the 6 miRNAs sponges and other 10 randomly selected ones were further validated to be circular by real-time PCR assay, and 14 displayed consistent regulation patterns with the transcriptome sequencing results. After Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the targeted mRNAs functions, the circRNAs were predicted to be involved in dehydration responsive process, such as photosynthesis, porphyrin, and chlorophyll metabolism, oxidative phosphorylation, amino acid biosynthesis, and metabolism, as well as plant hormone signal transduction, involving auxin, brassinosteroid, and salicylic acid. Herein, we revealed a possible connection between the regulations of circRNAs with the expressions of functional genes in wheat leaves associated with dehydration resistance.

[Repair mechanism of frozen sublethally damaged Staphylococcus aureus].

OBJECTIVE: To study the repair mechanisms of frozen sublethally damaged Staphylococcus aurous cells. METHODS: We resuscitated frozen sublethally damaged S. aureus at 37 degrees C for different time within 3 h. Meanwhile, we compared the morphological changes of the frozen sublethally damaged cells after 1 h of resuscitation using transmission electron microscopy assay (TEM). The expressions of the transcriptional attenuator MsrR (msrR), iron (Fe3+) ABC transporter ATP-binding protein (fhuC), and cytochrome b (cytB) genes were quantitatively analyzed by real-time fluorescence quantitative PCR (Real-time PCR) method. The content of cells outside leakage, active oxygen (ROS), and superoxide dismutase (SOD) activity were also determined by ultraviolet spectrophotometry. RESULTS: More than 99% of the frozen sublethally damaged S. aureus repaired after 3 h. The resuscitated cells expressed an equal resistance to high concentration of NaCl. Real-time PCR results showed that the msrR and fhuC genes expressions were down-regulated, whereas the cytB gene expression was up-regulated significantly. The frozen sublethally damaged S. aureus cellar surface ultrastructure significant changed during resuscitation. The cell surface became compact and sturdy from smooth and transparent. The cell leakage rate of ultraviolet absorption material gradually decreased. Meanwhile, the intracellular ROS level declined along with the decrease of SOD activity. CONCLUSION: Frozen sublethally damaged cells may regain the capability of resistance to high salt stress by repairing cell membrane integrity, reducing the content of ROS through gene regulation, inhibiting the toxicity of active oxygen to the cells. Meanwhile, the regulation of metabolism related genes (cytB) provides the energy for the requirement of cells, therefore, the frozen sublethally damaged cells were repaired finally.

Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide.

Real-time polymerase chain reaction (PCR) allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at -18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA) was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 103 CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 100 CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach.