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Pesticides introduced inadvertently or deliberately into environment by anthropogenic activity have caused growing global public concern, therefore the search of approaches for elimination of such xenobiotics should be encouraged. A cypermethrin-degrading bacterial strain Bacillus licheniformis B-1 was found to efficiently degrade carbaryl in LB medium at concentrations of 50–300 mg L−1 within 48 h, during which temperature and pH played important roles as reflected by increase in pollutant depletion. A stimulatory effect of Fe3+ and Mn2+ on microbial growth was observed, whereas Cu2+ caused inhibition of degradation. Results showed that 1-naphthol was a major transformation product of carbaryl which was further metabolised. An approximately 29 kDa carbaryl-degrading enzyme was purified from B-1 with 15.93-fold purification and an overall yield of 6.02% was achieved using ammonium sulphate precipitation, DEAE-Sepharose CL-6B anion-exchange chromatography and Sephadex G-100 gel filtration. The enzyme was identified through nano reversed-phase liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry as a phosphodiesterase (PDE). This is the first report on the characterization of carbaryl-degrading by Bacillus spp. and the role of a PDE in carbaryl-detoxifying. Also, strain B-1 showed versatile in carbosulfan, isoprocarb and chlorpyrifos degradation, demonstrating as ideal candidate for environment bioremediation.

The spoilage microbiology in Paocai (fermented vegetables) affects not only the quality of this popular traditional Chinese food but also its safety. This study aimed to isolate and identify the microorganisms commonly leading to spoilage in homemade Paocai from the Sichuan region and, further, to scientifically assess the impact of these microorganisms on product quality and safety. Seven putrid Paocai samples were collected from 7 families in different Sichuan cities. From these, 45 strains were isolated by means of a nutrient agar medium and rose bengal agar. All of the 22 strains (5 fungi and 17 bacteria) with different colonial morphologies and corruption phenomenon in Paocai were determined 16S rDNA/18S rDNA gene sequences. Bacteria were identified as Bacillus spp. (16 strains) and Staphylococcus saprophyticus (1 strain), while the 5 fungi were identified as Candida spp. (3 strains), Kodamaea ohmeri (1 strain), and Geotrichum candidum (1 strain). Based on the results of identification, 7 representative strains of different species were determined as the spoilage characteristics in paocai. All the representative strains metabolized to produce nitrite. Strain SPF21 (K. ohmeri) has a particularly serious impact on the crunchiness of Paocai; however, strains SPF19 (Bacillus subtilis) and SPF21 have the greatest influence on its color. All five of the fungi were seen to form a film on the surface of the Paocai, with SPF5 (G. candidum) exerting the most extreme influence. The growth characteristics in the broth showed that all the representative strains investigated metabolized most of the carbohydrates and were able to tolerate the salinity and acidity of the ordinary homemade Paocai. Moreover, these strains were found to have obvious impacts on the volatile components of Paocai, especially SPF2 and SPF8, with higher dimethyl disulfide and dimethyl trisulfide, which were found to have a pungent odor when highly concentrated.

A highly selective and effective method was successfully developed using magnetic molecular imprinted polymers (MMIPs) as solid-phase extraction (SPE) coupled with high performance liquid chromatography-ultraviolet detector (HPLC-UV) to rapidly determine cephalexin (CFX) in complex animal-derived food. MMIPs were creatively synthesized via suspension polymerization using Fe3O4 magnetic nanoparticles as supporter, CFX as template, acrylamide (AM) as functional monomer, and ethylene glycol dimethacrylate (EGDMA) as cross-linker. The MMIPs were characterized using X-ray diffraction (XRD), Fourier transform infrared spectrometry (FT-IR), scanning electron microscopy (SEM), and vibrating sample magnetometry (VSM). The binding process fitted well with pseudo-second-order model with good selectivity. Scatchard plot analysis suggested that MMIPs have two types of binding sites with the Qmax of 24.18 mg g−1 and 40.25 mg g−1, respectively. And Langmuir model proved that the recognition sites were uniformly distributed in a monolayer on the surface of MMIPs. The methodological assessment showed good applicability of MMIPs with excellent recovery (85.5%–94.0%), precision (1.2%–2.4%), and stability (intra-day 1.3%–3.6%; inter-day 2.6%–4.3%) in determining CFX content. In addition, the linearity of the calibration curve was good in the range of 0.02–5.00 mg L−1, with a sensitive detection limit of 5.00 μg kg−1. The results above suggest that the obtained MMIPs exert good performance for separation of CFX in animal-derived food, and the proposed method is suitable for the reliable determination of CFX in complex samples.

Vinegar is consumed worldwide as a food condiment, especially in the Chinese diet. The present study optimized the addition of A. niger biofortified-bran Qu (0.3%, 0.45%, and 0.6%) as additional starter to improve total acid content and starch utilization rate in industrial-scale Baoning vinegar production. In addition, this novel study determined the quality and microbial community changes of Baoning vinegar during three-round biofortification in industrial scale. Our results indicated that A. niger biofortified-bran Qu added at 0.6% resulted in higher total acid content and starch utilization rate of vinegar Pei. Biofortification imposed minor changes in the microbial community during three-round biofortification, and more variation was observed in fungal community than that in bacterial community. Most importantly, the quality of Baoning vinegar remained relatively stable. This information further confirmed the feasibility of multiple rounds of A. niger biofortification, and can be used to provide theoretical basis for industrial-scale production.

Cypermethrin (CY) is a synthetic pyrethroid widely used to control insect pests and it elicits a toxic effect on the human body. In this study, Bacillus licheniformis B-1 isolated from tea garden soil was used to degrade CY effectively. A specific enzyme was mainly localized in the extracellular compartments of B-1. This enzyme was identified as an esterase that could be produced without CY. The enzyme was purified 23.03-fold to apparent homogeneity with 8.38% overall recovery by ammonium sulfate precipitation, anion exchange chromatography, and gel filtration chromatography. The molecular mass of the CY-degrading enzyme was 66.4 kDa, and its optimal pH and temperature were 8.5 and 40 °C, respectively. Appropriate Zn2+ , Mn2+ , Mg2+ , Tween 80, SDS, Triton X-100, and BSA concentrations could greatly increase the activity of this enzyme. By contrast, EDTA, 1,10-phenanthroline, NaF, and PMSF strongly inhibited its activity. The purified enzyme showed Km and Vmax values were 5.532 nmol/mL and 33.445 nmol/min. The CY residue in lettuce and cherry tomatoes could be removed more than 50% under the conditions of the treatment concentration for 500 mg/L and the enzyme preparation dilution of 100 times. These results suggested that the CY-degrading enzyme, a constitutive enzyme that mainly exists in the extracellular space, was a novel esterase that might be used to detoxify CY, and could remove CY in vegetables effectively. PRACTICAL APPLICATION: Our research found a novel cypermethrin-hydrolyzing esterase from Bacillus licheniformis B-1 and proved that the enzyme could remove cypermethrin in vegetables effectively.

Background Lactiplantibacillus plantarum (L. plantarum), a dominant strain in traditional fermented foods, is widely used in fermentation industry because of its fast acid production. However, L. plantarum is easily inactivated due to acidity, high temperature and other factors. The formation of biofilm by bacteria can effectively increase environmental tolerance. Therefore, it is important to improve the environmental tolerance of L. plantarum by studying its biofilm formation conditions and regulatory mechanisms. Methods After determining a suitable NaCl concentration for promoting biofilm formation, L. plantarum was grown with 48 g L−1 NaCl. Differential gene expressions in L. plantarum biofilm vs. planktonic cells were analyzed using RNA sequencing and validated using qPCR. Result L. plantarum RS66CD biofilm formation formed highest amount of when grown at 48 g L−1 NaCl. Altogether 447 genes were up-regulated and 426 genes were down-regulated in the biofilm. KEGG pathway analysis showed that genes coding for D-Alanine metabolism, peptidoglycan biosynthesis, two-component system, carbon metabolism, bacterial secretion system, lysine biosynthesis and fatty acid metabolism were crucial for biofilm formation. In addition, eight other genes related to biofilm formation were differentially expressed. Our results provide insights into the differential gene expression involved in biofilm formation, which can help to reveal gene regulation during L. plantarum biofilm formation.

Cypermethrin is a common food contaminant and environmental pollutant that cause health threats to animals and humans. In this study, the characterization, mechanism, and application of cypermethrin removal by Saccharomyces cerevisiae were investigated. The binding of cypermethrin by the strains S. cerevisiae YS81 and HP was rapid and reached equilibrium at 2–8 h. The removal efficiency was dependent on incubation temperature and yeast concentration, whereas cypermethrin binding was not affected by pH. Heat and acid treatments enhanced the binding ability. Both strains survived in simulated digestion juices and removed cypermethrin effectively under simulated gastrointestinal conditions. Among the strains tested, the YS81 strain was the better candidate for cypermethrin concentration reduction. For the two S. cerevisiae strains, the biosorption kinetics and isotherm followed the pseudo-second-order model and Langmuir model well. The cell walls and the protoplasts were the main yeast cell components involved in cypermethrin binding. Fourier transformed infrared spectroscopy analysis revealed that –OH, –NH, –C–N, –COO−, and –C–O played a major role in binding cypermethrin. Inactive cells effectively removed cypermethrin from apple and cucumber juices and did not affect the physico-chemical properties. Therefore, S. cerevisiae strains YS81 and HP may be used for cypermethrin reduction in food or feed.

To improve the quality of Chinese traditional Paocai, two psychrotolerant lactic acid bacteria (LAB) strains were isolated from Paocai, and the quality of Chinese Paocai product using these two strains as starter cultures was compared to a control sample fermented with aged brine at 10 °C. The results suggested that the physicochemical and sensory features of Paocai fermented with psychrotolerant LAB were more suitable for industrial applications. The nitrite content of Paocai fermented with psychrotolerant LAB was 1 mg/kg, which was significantly lower than that of the control Paocai (P < 0.05). Low-temperature fermentation with the starter cultures of psychrotolerant LAB could effectively prevent overacidity and over-ripening of the Paocai products. Additionally, Paocai fermented with psychrotolerant LAB harbored relatively simple microbial flora as revealed by polymerase chain reaction-denaturing gradient gel electrophoresis. This study provides a basis for improving the quality of Chinese traditional Paocai and the large-scale production of low-temperature Chinese traditional Paocai products.

Biochemical oxygen demand (BOD) sensors based on Zr (IV)-loaded collagen fiber (ZrCF), a novel material with great porous structure, were developed. This novel material shows adsorbability by microorganisms. Saccharomyces cerevisiae and Escherichia coli were used for the construction of BOD sensors. Factors affecting BOD sensor performance were examined. The ZrCF-based BOD sensor showed different sensitivities and linear response ranges with different biofilm densities. The amount of microorganisms strongly affected the performance of the BOD sensor. Poor permeability of previously reported immobilization carriers were greatly circumvented by ZrCF. The service life of the ZrCF-based BOD sensor was more than 42 days. The immobilized microorganisms can be stored for more than 6 months under 4 °C in PB solution. There was good correlation between the results of the sensor method and the standard 5-day BOD method in the determination of pure organic substrates and real water samples.

Diethylstilbestrol (DES) can adversely affect the immune system of developing fetuses or even elicit toxic responses such as nerve toxicity and genotoxicity in human beings, thereby warranting methods to remove DES from the environments. The present study characterized a novel DES-degrading Bacillus subtilis JF and analyzed the degradation metabolites. The strain was collected at the China General Microbiological Culture Collection Center (Collection number: CGMCC 7950). The environmental effects, such as DES concentrations, pH levels, and temperature, on the strain’s degradation ability were tested. Degradation metabolites of DES by strain JF were analyzed via high performance liquid chromatography (HPLC) and liquid-chromatography time of flight mass spectrometry (LC-TOF-MS). Results indicated that B. subtilis JF can effectively degrade DES within a concentration of 25–200 mg/L. Increasing pH levels (pH > 7) are reported to increase the degradation rate of DES by the strain. The optimal temperature for strain JF to degrade DES was identified as 45°C. In this study, 4, 4′-hexene estrogen quinones (DESQ) and DES-4-semiquinone were speculated as two degradation metabolites of DES, and both can be completely degraded by strain JF. A slight reduction of DES in the blank system [DES cultured in Luria-Bertani (LB) medium without strain JF] was observed in this study. The reduction trend in the blank system only occurred during the first few days (about 4 days) and was considerably lesser than the decomposition and transformation effect of DES via strain JF. Furthermore, the metabolite DESQ could not be further decomposed in blank LB medium without strain JF. All the results demonstrate that complete degradation of DES in the fermentation broth occurs due to the function of strain JF rather than organic decomposition. In conclusion, the high efficiency of degradation and the potential to degrade DES completely indicates that strain JF has potential for the bioremediation of DES-contaminated environments (soil, river, and so on) and fermented foods.

Magnetic molecular imprinted polymers with ionic liquid used as an auxiliary solvent (IL@MMIPs) for the recognition of the methyl carbamate pesticide carbaryl (CBR) in foodstuff have been synthesized. The properties and application of IL@MMIPs were determined. The kinetic and isotherm adsorption processes were found to follow the pseudo-second-order and the Scatchard models, respectively. The selective experiment showed that the IL@MMIPs exhibited good selectivity to CBR compared to magnetic nonimprinted polymers with IL (IL@MNIPs). By using the IL@MMIPs as an adsorbent for the enrichment of CBR in food samples, the limit of detection (LOD, S/N = 3) and the limit of quantitation (LOQ, S/N = 10) of this method were 3 μg kg−1 and 10 μg kg−1, respectively. Compared with the traditional method, the IL@MMIP method has better recoveries (83.23–99.83%), precision (1.12–2.09%), and stabilization (intraday, 1.08–2.81%; interday, 2.26–3.30%). IL@MMIPs are an ideal adsorbent that could be applied to conveniently detect CBR in complex food, and the proposed method can be considered as a selective and sensitive alternative to traditional methods with affordable cost, avoiding the complex pretreatment procedure.

The authors have synthesized a phosphorescent probe of type SiO2-QD-MIP, where QD stands for Mn:ZnS quantum dots and MIP is a polymer coating that was molecularly imprinted with cephalexin. The nanoprobe with high specificity was prepared via sol-gel polymerization using thioglycolic acid (TGA)-modified QDs as luminescent materials, cephalexin as the template, 3-aminopropyltriethoxysilane as the functional monomer, and tetraethoxysilane as the crosslinking agent. The SiO2-QD-MIPs were characterized by X-ray powder diffraction, transmission electron microscopy, scanning electron microscopy, and Fourier transform infrared spectrometry. The orange emission of the probe, with excitation/emission maxima at 295/590 nm, decreases linearly in the 2.5–50 μg·L−1 cephalexin concentration range with a limit of detection (LOD) of 0.81 μg·L−1. The nanoprobe was successfully applied to the determination of cephalexin in (spiked) raw milk and milk powder. The recoveries ranged from 91.7 to 103.7%.

The purpose of this study was to evaluate the inhibitory mechanism of cell-free supernatants of Lactobacillus plantarum (LCFS) on Proteus mirabilis and estimate the expression of histamine synthesis-related genes. The minimum inhibitory concentration (MIC) of the LCFS and antibacterial curve assay was determined by the tube double-dilution method. The antibacterial mechanism was explored by cell integrity, morphology, and structure of P. mirabilis. Further, the expression of the histidine decarboxylase (hdcA) gene and histidine/histamine antiporter (hdcP) gene of P. mirabilis and the intracellular and extracellular contents of histidine and histamine were determined. The MIC of the LCFS was 12.5 mg/mL, and the antibacterial curve assay showed an inhibited growth of P. mirabilis. The increase in the Alkaline Phosphatase (AKP) content, membrane permeability, and potential indicated the destruction of the cytoplasm, cell membrane, and cell wall of P. mirabilis. The results of SEM and TEM revealed that the LCFS disrupted the normal morphology and structure of the cell. Moreover, differences in the extracellular and intracellular histidine and histamine content between the MIC group and the control group was consistent with the decreased expression of hdcA and hdcP genes in the MIC group observed in the previous antimicrobial studies. Hence, the inhibitory mechanism of LCFS on histamine formation was clarified at cellular and molecular levels.

In this study, the cypermethrin binding characteristics of lactic acid bacteria were investigated for the first time. Two strains, Lactobacillus plantarum RS60 and Pediococcus acidilactici D15, possessed the highest cypermethrin removal capacity and good tolerance to simulated digestive juices. They were employed for further studies on cypermethrin binding characteristics. 55.06% and 56.46% of cypermethrin were removed within 0.25 h by strains RS60 and D15, respectively. The effect of pH on binding capacity was negligible. Heat treatment enhanced cypermethrin binding rate. Moreover, inactive cells were capable of removing cypermethrin from fruit and vegetable juices, with over 60% cypermethrin reduction within 2 h. No adverse effect was found on the quality of juice during the biosorption process. Besides, these two strains also could bind other several pyrethroids and 3-phenoxybenzoic acid. These findings indicated that L. plantarum RS60 and P. acidilactici D15 may be useful to reduce cypermethrin in contaminated foods.

This paper reports the development of a novel probe based on magnetic room-temperature phosphorescence quantum dots with molecularly imprinted polymers (MQD-MIPs) for the rapid detection of trace norfloxacin (NFX) residual in complex food matrix. The highly selective probe was constructed by surface molecular imprinting technology using magnetic materials (Fe3O4 nanoparticles) as core, Mn-doped ZnS quantum dots (Mn-ZnS QDs) as phosphorescent materials, NFX as template, 3-aminopropyltriethoxysilane as functional monomer, and tetraethoxysilane as crosslinking agent. The as-obtained MQD-MIPs were characterized in detail by transmission electron microscopy, scanning electron microscopy, X-ray powder diffraction, Fourier transform infrared spectrometry, and vibrating sample magnetometer. A magnetic strength of 37.64 emu g−1 was recorded. Also, the probe displayed excellent room temperature phosphorescence properties with excitation/emission peaks at 300/590 nm. Under the optimized conditions, the detection time was less than 40 min, phosphorescence intensity varied linearly with concentration from 1 to 90 μg·L−1, and detection limit reached as low as 0.80 μg·L−1. Furthermore, the MQD-MIPs-based probe successfully detected norfloxacin residues in spiked fish and milk samples with recoveries of 90.92–111.53% and RSD

To further improve the utilization rate of starch in the fermentation of Baoning vinegar, the effects of Aspergillus niger biofortification on the quality and microbial community of vinegar Pei (a mixture of fermentation materials) and Baoning vinegar were evaluated. The results suggested that the total acid content, a crucial parameter for vinegar production, in vinegar Pei increased by approximately 1.70 g/100 g at the end of fermentation. The utilization rate of starch and content of organic acid increased significantly after biofortification, and the content of lactic acid was the highest in Baoning vinegar, followed by acetic acid. Similarly, the total amount of free amino acids increased after biofortification, especially the content of histidine and tyrosine. The esters and acid compounds were the most abundant in the vinegar Pei, while the contents of acid and carbonyl compounds were the highest in liquid vinegar. As revealed by high-throughput sequencing, the microbial community in vinegar Pei changed little after biofortification. Lactic acid bacteria and yeast are the most abundant microbes throughout the fermentation process. The present study provides insights into the improvement of the solid-state brewing process of vinegar.

The strain Y1, with a notably high production of neutral protease, was isolated from naturally fermented broad beans and subsequently identified as Aspergillus oryzae, through the analysis of its morphology characteristics and 18S rDNA sequence. Naturally fermented broad beans are the main raw material in Sichuan broad-bean sauce. The neutral protease from Aspergillus oryzae Y1 was purified using ammonium sulphate precipitation and DEAE-Sepharose Fast Flow chromatography, which resulted in a 10.0-fold increase in the specific activity (2264.3 U/mg) and a recovery rate of 21%. The estimated molecular mass of the purified protease was approximately 45 kDa. The optimal pH and temperature of the purified protease were 7.0 and 55 °C, respectively. The heat resistance of the purified protease was significantly higher than the commercial protease. The effect of metal ions on the activity of the purified protease approximated that of commercial neutral protease. Furthermore, the maximum hydrolysis rate (V max) and apparent Michaelis–Menten constant (K m) values of the purified protease were 256.4103 μg/mL min and 20.0769 mg/mL, respectively. The purified protease had a higher affinity for the substrate than the commercial neutral protease. All the results suggest that this neutral protease exhibits the potential for application in industry due to its good resistance to high temperatures and wide range of acids and bases.

In this study, an efficient, selective, and simple analytical method for the extraction of chloramphenicol (CAP) from food using magnetic molecularly imprinted polymers (MMIPs) as the solid-phase extraction (SPE) sorbent was successfully developed. MMIPs with varying ratios of methacrylic acid to acrylamide were prepared by suspension polymerization on the surface of double-bond-modified Fe3O4 magnetic nanoparticles. Further, these MMIPs were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy, as well as using a vibrating sample magnetometer. Furthermore, the adsorption capacities of MMIPs and MNIPs were investigated by binding experiments. Methodology evaluation for the detection of CAP from food was carried out using MMIPs as the SPE sorbent. By using an external magnetic field, MMIPs were separated by a simple and rapid method. The diameter of the so-obtained MMIPs, exhibiting good monodispersity, was 400–700 nm. The MMIPs exhibited the maximum apparent adsorption capacity of up to 42.60 mg g−1 with good selectivity. For the detection of food samples, the linear response range was 0.02–10.00 mg L−1, with a detection limit of 10 μg L−1, and intra- and inter-day stabilities ranged from 1.34% to 1.89% and from 1.76% to 2.77%, respectively, with good recoveries (95.31%–106.89%) and satisfactory relative standard deviations (1.21%–2.60%).

Chicken products with high water and nutrient contents provide essential nutrients for microbial growth. This study was designed to determine the effect of lactic acid-based spray washing. First, the reduction of microbial counts on chicken carcasses was evaluated through lactic acid-based spray washing. Second, bacterial diversity on chicken carcasses in different steps in a chicken-slaughtering chain was investigated through polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The effect of spray washing on quick-frozen chicken quality was also examined. Results revealed that microbial counts decreased significantly after the chickens were spray-washed with 1.5% lactic acid at 50 °C for 15s. PCR-DGGE revealed a decrease in microbial counts of several bacterial species, such as uncultured bacterium, uncultured Clostridiales sp., Pseudomonas fluorescens , Ruminococcus sp., and uncultured Ruminococcus sp. The spray washing performed did not remarkably affect the pH and total volatile basic nitrogen of quick-frozen chicken during storage. Therefore, the proposed technique may be applied to chicken-slaughtering chains and may be used to investigate bacterial diversity on chicken carcasses.

We extracted collagen (CL) from the scales of Ctenopharyngodon idella and fabricated a CL self-assembly film. Catalase (CAT) was immobilized on the films using cross-linking, adsorption, and embedding methods. The activity and operational stability of immobilized CAT were investigated, along with the influence of the concentration of glutaraldehyde (GTA) and of the initial concentration of embedded CAT. The results showed that the CL triple helix remained intact. Differential scanning calorimetry data showed that the thermal stability of CL was significantly improved by neutral salt-induced self-aggregation and GTA cross-linking. The immobilized enzyme had high activity and good operational stability. When the enzyme concentration reached 0.5 mg/mL, immobilized enzyme activity of grass carp scales reached a maximum of 2596 U/g, and after twenty-two uses, enzyme activity remained above 50%, and it could be reused >45 times (CAT = 0.5 mg/mL, GTA = 5%, Temperature = 30 °C, pH = 7). Moreover, the optimum temperature and pH of immobilized CAT were 35 °C and 7, respectively, while the same for free CAT was 30 °C and 7, respectively. This indicated that immobilization of CAT has a protective effect.

The exopolysaccharides (EPS) produced by Se-enriched Lactobacillus plantarum BC-25 was purified to illustrate its structure and conformational characterization. The yield of EPS (324.80mg/l) was obtained with a sodium selenite concentration of 6μg/ml. The results indicated that the EPS was soluble in water, but insoluble in organic solvents. The molecular weight of this highly thermal stability EPS was 1.83×10(4)Da and 1.33×10(4)Da with or without Se enriched respectively. The EPS was composed of mannose, galactose and glucose in a molar ratio of 92.21:1.79:6.00 and 91.36:2.44:6.20 with or without Se. This compound had a backbone of (1→2)-linked Man, (1→2.6)-linked Glc, (2→6)-linked Man, and (2→6)-linkedGal confirmed by GC-MS. IR analysis suggested that the EPS belonged to heteropolysaccharide with a pyran group, with possible presence of SeO and CSeC residues that Se may substitutes CH3 in -OCH3 in the polysaccharide as confirmed by NMR spectroscopy.