Microwave-Triggered 4D Automatic Color Change in 3D-Printed Food Materials Incorporating Natural Pigments.

The feasibility of using microwaves to quickly stimulate automatic color change in 3D-printed food containing curcumin or anthocyanins was studied. Firstly, with a dual-nozzle 3D printer, stacked structures included mashed potatoes (MPs, upper part, containing anthocyanins) and lemon juice-starch gel (LJSG, lower part) were 3D-printed and post-treated using a microwave. The results indicated that the viscosity and gel strength (indicated by the elastic modulus (G') and complex modulus (G*)) of LJSG were improved with the increase in starch concentration, while water mobility was reduced. During microwave post-treatment, the color change speed was negatively correlated with the gel strength but positively correlated with the diffusion of H+ and anthocyanin concentration. Secondly, nested structures were 3D-printed using MPs containing curcumin emulsion and baking soda (NaHCO3). During microwave post-treatment, the curcumin emulsion structure was destroyed, and NaHCO3 was decomposed, along with an increase in alkalinity; thus, the automatic color change was achieved with the automated presentation of hidden information. This study suggests that 4D printing could enable the creation of colorful and attractive food structures using a household microwave oven, leading to more imaginative solutions regarding personalized foods, which may be particularly important to people with poor appetites.

Rapid determination of 2,4-diaminopyrimidine residues through sample pretreatment using immunomagnetic bead purification along with HPLC-UV.

To reduce matrix interference and realize simultaneous detection of multiple homologous compounds (trimethoprim (TMP), diaveridine (DVD), ormetoprim (OMP), baquiloprim (BQP), and aditoprim (ADP) in pig, cattle, chicken, and fish muscles), an immunomagnetic bead (IMB)-based sample purification pretreatment with HPLC-UV was developed. A broad-spectrum monoclonal antibody (mAb, named 14C6) was prepared and conjugated with carboxylic-acid-functionalized magnetic nanoparticles using the active ester method to obtain IMBs for sample purification. The extraction solvent was optimized based on the extraction efficiency. Good linearity was observed for all the five analytes (10-200 µg/kg) with the LOD and LOQ of 5 and 10 µg/kg, respectively. The mean recoveries ranged from 62.5% to 76.9%, while the coefficient of variation was <12.2%. The IMB method afforded greater sample purification and enrichment than those achieved with the SPE column-based conventional method. Hence, the IMB-based sample purification is a useful tool to determine 2,4-diaminopyrimidine residues in edible animal tissues.

Quorum Sensing-Mediated and Growth Phase-Dependent Regulation of Metabolic Pathways in Hafnia alvei H4.

Quorum sensing (QS) is a widespread regulatory mechanism in bacteria used to coordinate target gene expression with cell density. Thus far, little is known about the regulatory relationship between QS and cell density in terms of metabolic pathways in Hafnia alvei H4. In this study, transcriptomics analysis was performed under two conditions to address this question. The comparative transcriptome of H. alvei H4 wild-type at high cell density (OD600 = 1.7) relative to low cell density (OD600 = 0.3) was considered as growth phase-dependent manner (GPDM), and the transcriptome profile of luxI/R deletion mutant (ΔluxIR) compared to the wild-type was considered as QS-mediated regulation. In all, we identified 206 differentially expressed genes (DEGs) mainly presented in chemotaxis, TCA cycle, two-component system, ABC transporters and pyruvate metabolism, co-regulated by the both density-dependent regulation, and the results were validated by qPCR and swimming phenotypic assays. Aside from the co-regulated DEGs, we also found that 59 DEGs, mediated by density-independent QS, function in pentose phosphate and histidine metabolism and that 2084 cell-density-dependent DEGs involved in glycolysis/gluconeogenesis and phenylalanine metabolism were influenced only by GPDM from significantly enriched analysis of transcriptome data. The findings provided new information about the interplay between two density-dependent metabolic regulation, which could assist with the formulation of control strategies for this opportunistic pathogen, especially at high cell density.

Lentibacillus panjinensis sp. nov., Isolated from Shrimp Paste, a Traditional Chinese Fermented Seafood.

A Gram-positive, aerobic, motile and short rod-shaped bacterium, designated as strain G56T, was isolated from shrimp paste produced in Panjin, China. Grows in the presence of 1.0-25.0% (w/v) NaCl (optimum at 10%), pH 5.0-9.5 (optimally at 7.0) and 10-50 °C (optimally at 37 °C). Positive for catalase and oxidase activities, but lack the ability to reduce nitrate. Acids produce from D-ribose, D-xylose, D-galactose, glycerol and D-trehalose, but no acid is produced when salicin, D-mannose, D-cellobiose and L-sorbose are provided as substrates. The polar lipid extract is found to contain diphosphatidylglycerol, phosphatidylglycerol, an unknown glycolipid, and unidentified phospholipids. Fatty acids are mainly defined as anteiso-C15:0 (69.7%) and anteiso-C17:0 (23.3%). The G+C content of its DNA is 44.7 mol%. The draft genome of strain G56T is 3,209,087 bp in length and the average nucleotide identity value (ANI) and the digital DNA-DNA hybridization (DDH) values between strain G56T and L. juripiscarius JCM 12147T is 78.41% and 22.0%, respectively. Polyphasic taxonomic analysis classified strain G56T as a novel species in the genus Lentibacillus, and therefore, we named it as Lentibacillus panjinensis sp. nov..

Effect of quorum sensing and quorum sensing inhibitors on the expression of serine protease gene in Hafnia alvei H4.

The serp gene codes for a protease that is considered to be an important factor associated with quorum sensing (QS)-based food spoilage caused by microorganisms. In this study, we evaluated the effect of temperature (4-37 °C) and QS inhibitors on the production of N-acyl-L-homoserine lactones (AHLs) and relative expression of the luxR/I, as well as serp in Hafnia alvei H4. Production of AHLs and expression of luxR/I were found to reach maximum levels at 10 °C, suggesting that the QS system of H. alvei H4 might have higher activity at low temperatures; similar result was also obtained for serp expression. Mutants of H. alvei H4 deficient in QS were used to identify the regulation of QS on serp expression. Significant reduction (P < 0.05) in serp expression was found in the mutants ∆luxR, ∆luxI, and ∆luxR/I, with ∆luxI and ∆luxR/I showing greater reduction than ∆luxR. Minimum inhibition concentrations (MIC) of Benzyl isothiocyanate and 3-Methylthiopropyl isothiocyanate for H. alvei H4 were determined to be 7.813 and 15.625 mM, respectively. Furthermore, the expression of serp, as well as that of luxR and luxI, was significantly repressed (P < 0.05) by the two QS inhibitors at 1/8 MIC and 1/16 MIC, indicating that these inhibitors might repress serp expression through affecting luxR and luxI expression in H. alvei H4. The findings of this study, therefore, suggested that food spoilage caused by H. alvei could be controlled through the application of QS inhibitors.

The Impact of Microbial Diversity on Biogenic Amines Formation in Grasshopper Sub Shrimp Paste During the Fermentation.

Biogenic amines (BAs) and microbial diversity are important factors affecting food quality and safety in fermented foods. In this study, the bacterial and fungal diversity in grasshopper sub shrimp paste taken at different fermentation times were comprehensively analyzed, while the pH, colony counts, salinity, total volatile base nitrogen (TVB-N) and BA contents were quantitatively determined. In addition, the correlations among the samples with respect to microbial communities and the different parameters investigated especially BAs were also established. By combining the results of spearman correlation heatmap with the contents of BAs produced by the 102 halotolerant bacteria isolated from the grasshopper sub shrimp paste, six major genera of bacteria (Jeotgalibaca, Jeotgalicoccus, Lysinibacillus, Sporosarcina, Staphylococcus, and Psychrobacter) were found to be positively correlated with BA production level, suggesting that these bacteria might have a strong tendency to produce BAs. Other bacteria such as Lentibacillus, Pseudomonas, and Salinicoccus were considered as poor BA producers. The grasshopper sub shrimp paste was characterized by a relatively high abundance of Tetragenococcus, which was the dominant genus during the fermentation process, and it also produced a relatively high level of BAs but the spearman correlation heatmap revealed a negative correlation between T. muriaticus and BA level. Analysis of the species relevance network in grasshopper sub shrimp explained that the actual production of BAs by a certain strain was closely related to other species present in the complex fermentation system.

Effect of the luxI/R gene on AHL-signaling molecules and QS regulatory mechanism in Hafnia alvei H4.

Hafnia alvei H4 is a bacterium subject to regulation by a N-acyl-l-homoserine lactone (AHL)-mediated quorum sensing system and is closely related to the corruption of instant sea cucumber. Studying the effect of Hafnia alvei H4 quorum sensing regulatory genes on AHLs is necessary for the quality and preservation of instant sea cucumber. In this study, the draft genome of H. alvei H4, which comprises a single chromosome of 4,687,151 bp, was sequenced and analyzed and the types of AHLs were analyzed employing thin-layer chromatography (TLC) and high resolution triple quadrupole liquid chromatography/mass spectrometry (LC/MS). Then the wild-type strain of H. alvei H4 and the luxI/R double mutant (ΔluxIR) were compared by transcriptome sequencing (RNA-seq). The results indicate that the incomplete genome sequence revealed the presence of one quorum-sensing (QS) gene set, designated as lasI/expR. Three major AHLs, N-hexanoyl-L-homoserine lactone (C6-HSL), N-butyryl-L-homoserine lactone (C4-HSL), and N-(3-oxo-octanoyl)-L-homoserine lactone (3-oxo-C8-HSL) were found, with C6-HSL being the most abundant. C6-HSL was not detected in the culture of the luxI mutant (ΔluxI) and higher levels of C4-HSL was found in the culture of the luxR mutant (ΔluxR), which suggested that the luxR gene may have a positive effect on C4-HSL production. It was also found that AHL and QS genes are closely related in the absence of luxIR double deletion. The results of this study can further elucidate at the genetic level that luxI and luxR genes are involved in the regulation of AHL.

Characteristics of N-Acylhomoserine Lactones Produced by Hafnia alvei H4 Isolated from Spoiled Instant Sea Cucumber.

This study aimed to identify N-acylhomoserine lactone (AHL) produced by Hafnia alvei H4, which was isolated from spoiled instant sea cucumber, and to investigate the effect of AHLs on biofilm formation. Two biosensor strains, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens KYC55, were used to detect the quorum sensing (QS) activity of H. alvei H4 and to confirm the existence of AHL-mediated QS system. Thin layer chromatography (TLC) and high resolution triple quadrupole liquid chromatography/mass spectrometry (LC/MS) analysis of the AHLs extracted from the culture supernatant of H. alvei H4 revealed the existence of at least three AHLs: N-hexanoyl-l-homoserine lactone (C6-HSL), N-(3-oxo-octanoyl)-l-homoserine lactone (3-oxo-C8-HSL), and N-butyryl-l-homoserine lactone (C4-HSL). This is the first report of the production of C4-HSL by H. alvei. In order to determine the relationship between the production of AHL by H. alvei H4 and bacterial growth, the ß-galactosidase assay was employed to monitor AHL activity during a 48-h growth phase. AHLs production reached a maximum level of 134.6 Miller unites at late log phase (after 18 h) and then decreased to a stable level of about 100 Miller unites. AHL production and bacterial growth displayed a similar trend, suggesting that growth of H. alvei H4 might be regulated by QS. The effect of AHLs on biofilm formation of H. alvei H4 was investigated by adding exogenous AHLs (C4-HSL, C6-HSL and 3-oxo-C8-HSL) to H. alvei H4 culture. Biofilm formation was significantly promoted (p < 0.05) by 5 and 10 µM C6-HSL, inhibited (p < 0.05) by C4-HSL (5 and 10 µM) and 5 µM 3-oxo-C8-HSL, suggesting that QS may have a regulatory role in the biofilm formation of H. alvei H4.