Effect of Lacticaseibacillus casei LC2W Supplementation on Glucose Metabolism and Gut Microbiota in Subjects at High Risk of Metabolic Syndrome: A Randomized, Double-blinded, Placebo-controlled Clinical Trial.

Metabolic syndrome (MetS) is a global epidemic complex and will cause serious metabolic comorbidities without treatment. A prevention strategy for MetS development has been proposed to modulate gut microbiota by probiotic administration to improve intestinal dysbiosis and benefit the host. Lacticaseibacillus casei LC2W has exhibited positive effects in preventing colitis and anti-hypertension in vivo. However, the effect of L. casei LC2W on subjects at high risk of MetS is unknown. Here, a randomized, double-blinded, placebo-controlled study was conducted on 60 subjects with high risk of MetS, and the hypoglycemic and hypolipidemic activity and possible pathways of L. casei LC2W were inferred from the correlation analysis with gut microbiome composition, function, and clinical phenotypic indicators. The results showed that oral administration of L. casei LC2W could exert significant benefits on weight control, glucose and lipid metabolism, inflammatory and oxidative stress parameters, and SCFA production, as well as modulate the composition of gut microbiota. The relative abundance of Lacticaseibacillus, Bifidobacterium, Dorea, and Blautia was enriched, and their interaction with other gut microbes was strengthened by oral administration of L. casei LC2W, which was beneficial in ameliorating gut inflammation, promoting glucose and lipids degradation pathways, thus alleviated MetS. The present study confirmed the prevention effects of L. casei LC2W towards MetS from aspects of clinical outcomes and microflora modulation, providing an alternative strategy for people at high risk of MetS.Trial registration: The study was proactively registered in ClinicalTrial.gov with the registration number of ChiCTR2000031833 on April 09, 2020.

Lactiplantibacillus plantarum ST-III and Lacticaseibacillus rhamnosus KF7 Enhance the Intestinal Epithelial Barrier in a Dual-Environment In Vitro Co-Culture Model.

Intestinal barrier hyperpermeability, which is characterised by impaired tight junction proteins, is associated with a variety of gastrointestinal and systemic diseases. Therefore, maintaining intestinal barrier integrity is considered one of the effective strategies to reduce the risk of such disorders. This study aims to investigate the potential benefits of two probiotic strains (Lactiplantibacillus plantarum ST-III and Lacticaseibacillus rhamnosus KF7) on intestinal barrier function by using a physiologically relevant in vitro model of the intestinal epithelium. Our results demonstrate that both strains increased transepithelial electrical resistance, a measure of intestinal barrier integrity. Immunolocalisation studies indicated that this improvement in barrier function was not due to changes in the co-localisation of the tight junction (TJ) proteins ZO-1 and occludin. However, we observed several modifications in TJ-related genes in response to the probiotics, including the upregulation of transmembrane and cytosolic TJ proteins, as well as TJ signalling proteins. Gene expression modulation was strain- and time-dependent, with a greater number of differentially expressed genes and higher fold-change being observed in the L. plantarum ST-III group and at the latter timepoint. Further studies to investigate how the observed gene expression changes can lead to enhanced barrier function will aid in the development of probiotic foods to help improve intestinal barrier function.

Recent Studies and Applications of Hydrogel-Based Biosensors in Food Safety.

Food safety has increasingly become a human health issue that concerns all countries in the world. Some substances in food that can pose a significant threat to human health include, but are not limited to, pesticides, biotoxins, antibiotics, pathogenic bacteria, food quality indicators, heavy metals, and illegal additives. The traditional methods of food contaminant detection have practical limitations or analytical defects, restricting their on-site application. Hydrogels with the merits of a large surface area, highly porous structure, good shape-adaptability, excellent biocompatibility, and mechanical stability have been widely studied in the field of food safety sensing. The classification, response mechanism, and recent application of hydrogel-based biosensors in food safety are reviewed in this paper. Furthermore, the challenges and future trends of hydrogel biosensors are also discussed.

Lactiplantibacillus plantarum ST-III-fermented milk improves autistic-like behaviors in valproic acid-induced autism spectrum disorder mice by altering gut microbiota.

Introduction: Autism spectrum disorder (ASD) is a serious neurodevelopmental disorder with a rising incidence. More and more studies have shown that abnormal microbiota composition may aggravate the behavioral symptoms and biological signs of ASD, and interventions of probiotics and diet have emerged as a potential improvement measure. Methods: Lactiplantibacillus plantarum ST-III-fermented milk was applied as an oral intervention in a valproic acid (VPA)-induced ASD mice model, and the effect of probiotic intake on autistic-related behaviors and gut microbiota composition was evaluated by behavioral tests and 16S rRNA gene sequencing. Results: Gender specificity was shown in VPA-induced behavioral abnormalities in a mouse model, and L. plantarum ST-III-fermented milk was effective in ameliorating the impaired social interaction in male ASD mouse models, but not for the anxiety behavior exhibited by female ASD mouse models. Meanwhile, dietary changes were found to be the main cause of the altered gut microbiota in mice, and additional intake of L. plantarum ST-III-fermented milk seemed to improve autistic-like behaviors in male ASD mouse models by modulating specific gut microbes. Discussion: These findings suggest that L. plantarum ST-III-fermented milk may play a beneficial role in improving the behavioral symptoms of ASD and is expected to be one of the candidate functional foods for ASD.

Lactobacillus plantarum ST-III modulates abnormal behavior and gut microbiota in a mouse model of autism spectrum disorder.

Probiotic treatment might improve autism spectrum disorder (ASD) behavior. In this study, we investigated the improvement effects of Lactobacillus plantarum ST-III on a mouse model of ASD, which was constructed using triclosan. After two weeks of L. plantarum ST-III oral feeding, autism-like social deficits in male mouse models were ameliorated. L. plantarum ST-III also attenuated the self-grooming and freezing times of female mice. High-throughput sequencing revealed changes in the gut microbiota after L. plantarum ST-III intervention. In the female probiotic group, the abundance of beneficial Lachnospiraceae bacteria increased, whereas that of harmful Alistipes bacteria decreased. Correlation analysis showed that amelioration of abnormal behavior in a mouse model of ASD was related to the involvement of certain metabolic pathways. A reduction in the abundance of Alistipes was involved in stereotyped behavioral improvement. Thus, oral supplementation with L. plantarum ST-III can help improve social behavior in a male mouse model of ASD and contribute to more balanced intestinal homeostasis.

Endophytic Diaporthe Associated with Morinda officinalis in China.

Diaporthe species are endophytes, pathogens, and saprobes with a wide host range worldwide. However, little is known about endophytic Diaporthe species associated with Morinda officinalis. In the present study, 48 endophytic Diaporthe isolates were obtained from cultivated M. officinalis in Deqing, Guangdong Province, China. The nuclear ribosomal internal transcribed spacer (ITS), partial sequences of translation elongation factor 1-α (tef1-α), partial calmodulin (cal), histone H3 (his), and Beta-tubulin (ß-tubulin) gene regions were sequenced and employed to construct phylogenetic trees. Based on morphology and combined multigene phylogeny, 12 Diaporthe species were identified, including five new species of Diaporthe longiconidialis, D. megabiguttulata, D. morindendophytica, D. morindae, and D. zhaoqingensis. This is the first report of Diaporthe chongqingensis, D. guangxiensis, D. heliconiae, D. siamensis, D. unshiuensis, and D. xishuangbanica on M. officinalis. This study provides the first intensive study of endophytic Diaporthe species on M. officinalis in China. These results will improve the current knowledge of Diaporthe species associated with this traditional medicinal plant. Furthermore, results from this study will help to understand the potential pathogens and biocontrol agents from M. officinalis and to develop a disease management platform.

Lasiodiplodia fici sp. nov., Causing Leaf Spot on Ficus altissima in China.

High temperatures and the seasonality in tropical ecosystems favours plant pathogens, which result in many fungal diseases. Among these, diseases caused by Botryosphaeriaceae species are prominent as dieback, canker and leaf spots. In this research, we isolated one leaf-spot-causing Botryosphaeriaceae species from Ficus altissima leaves, which were collected in Guangzhou, Guangdong Province, China. Isolation and identification of the pathogen were based on morphological and molecular aspects. Based on multigene phylogenetic analysis of combined internal transcribed spacer (ITS), translation elongation factor 1-α gene (tef1) and beta-tubulin gene (tub2), the fungus associated with leaf spots on F. altissima is described as Lasiodiplodia fici, a novel species. Pathogenicity assays were conducted by inoculating the fungus onto detached shoots and plants under controlled environmental conditions. The results revealed that the L. fici isolates can infect the plant tissues under stress conditions by developing disease symptoms on detached shoots within three days. However, when it was inoculated onto the leaves of the host and grown in natural conditions, the progression of the disease was slow. The putative pathogen was re-isolated, and Koch's assumptions were satisfied. This is the first report of Lasiodiplodia species causing disease on Ficus altissima. Results from the present study will provide additional knowledge on fungal pathogens associated with forest and ornamental plant species.

16S ribosomal RNA-depletion PCR and its application in cause analysis of yogurt package shrinkage.

Fermentative bacteria, the main microbiota in yogurt, interfere with the detection of unintended bacterial contaminants. The removal of fermentative bacteria and enrichment of unintended bacterial contaminants is a challenging task in bacterial detection. The present study developed a new 16S rRNA-depletion PCR for such enrichment and detection. Specifically, a single-guide RNA was designed and synthesized based on the 16S rRNA sequence of Streptococcus thermophilus, with the highest DNA abundance in the yogurt. The CRISPR-Cas9 system was used to specifically cleave and remove the genomic DNA of the fermentative bacteria, followed by PCR amplification. This method improved the detection sensitivity, simplified the operation steps, and reduced the detection cost of PCR analysis. We also used the 16S rRNA-depletion PCR to amplify and detect the unintended bacterial contaminants in yogurts with shrunken packages and analyzed the underlying reasons to prevent this issue of product quality.

Co-infection of Fusarium aglaonematis sp. nov. and Fusarium elaeidis Causing Stem Rot in Aglaonema modestum in China.

Aglaonema modestum (A. modestum) (Araceae) is an evergreen herbage, which is intensively grown as an ornamental plant in South China. A new disease was observed in A. modestum from 2020 to 2021 in Guangdong province, China. The disease symptoms associated with plants were initial leaf wilt, stem rot, and resulting plant death, leading to severe economic losses. In total, six Fusarium isolates were obtained from diseased plants. The putative pathogen was identified using both morphological characteristics and molecular phylogenetic analysis of calmodulin A (cmdA), RNA polymerase largest subunit 1 (rpb1), RNA polymerase II (rpb2), translation elongation factor-1α (tef1-α), and beta-tubulin (ß-tubulin) sequences. Two Fusarium species were identified, namely, one new species, Fusarium aglaonematis (F. aglaonematis) belonging to Fusarium fujikuroi species complex. In addition, Fusarium elaeidis (F. elaeidis) belonging to the Fusarium oxysporum (F. oxysporum) species complex was also identified. Pathogenicity assays were conducted by inoculating each species into potted A. modestum plants and co-inoculating two species. The results showed that two Fusarium species could infect plants independently and can infect them together. Co-infection of these two species enhanced the disease severity of A. modestum. Compared to single inoculation of F. elaeidis, severity was higher and disease development was quicker when plants were only inoculated with F. aglaonematis. In addition, these two Fusarium species could infect Aglaonema plants without wounds, while inoculation with a physical injury increased disease severity. This is the first report of co-infection by F. aglaonematis and F. elaeidis causing stem rot on A. modestum worldwide. This study will be an addition to the knowledge of Fusarium diseases in ornamental plants. These results will provide a baseline to identify and control diseases associated with A. modestum.

Determination of 5-Hydroxymethylfurfural (5-HMF) in milk products by surface-enhanced Raman spectroscopy and its simulation analysis.

5-Hydroxymethylfurfural (5-HMF) is a useful indicator of thermal damage degree and freshness of milk. It is of great importance to develop simple, rapid and accurate analytical methods for the sensitive detection of 5-HMF in milk and milk-based products. In this work, surface-enhance Raman spectroscopy (SERS) was used for rapid determination of 5-HMF in processed cheese by colloidal Au nanoparticles (AuNPs) substrate synthesized by the classical solvothermal reduction method. Density functional theory (DFT) calculations were carried out to determine the vibration assignments of 5-HMF and the surface enhancement effect of AuNPs substrate. The results found that a good linear response on the AuNPs substrate for 5-HMF in the concentration range of 0.1-75 mM was established with the detection limit of 75 µM (S/N = 3). Furthermore, the present method could be applied to the determination of 5-HMF in a cheese real sample which revealed its promising application in food safety and analysis.

Lactobacillus paracasei BD5115-Derived 2-Hydroxy-3-Methylbutyric Acid Promotes Intestinal Epithelial Cells Proliferation by Upregulating the MYC Signaling Pathway.

Metabolites of probiotics that are beneficial to human health have been isolated from the intestinal tract and natural dairy products. However, many studies on probiotics and prebiotics are limited to the observation of human cohorts and animal phenotypes. The molecular mechanisms by which metabolites of probiotics regulate health are still need further exploration. In this work, we isolated a strain of Lactobacillus Paracasei from human milk samples. We numbered it as Lactobacillus Paracasei BD5115. The mouse model of high-fat diet confirmed that the metabolites of this strain also promotes intestinal epithelial cells (IECs) proliferation. Single-cell sequencing showed that a bZIP transcription factor MAFF was specifically expressed in some IECs. We found that MAFF interacted with MBP1 to regulate the expression of MYC. Analysis of the active components in BD5115 metabolites confirmed that 2-hydroxy-3-methylbutyric acid promotes the expression of the MYC gene. This promotes the proliferation of IECs. Our findings indicate that 2-hydroxy-3-methylbutyric acid regulate MYC gene expression mediated by MAFF/MBP1 interaction. This study not only screened a strain with promoted IECs proliferation, but also discovered a new signal pathway that regulates MYC gene expression.

Fusarium elaeidis Causes Stem and Root Rot on Alocasia longiloba in South China.

Alocasia longiloba is a popular ornamental plant in China, however pests and diseases associated with A. longiloba reduce the ornamental value of this plant. From 2016 to 2021, stem and root rot has been observed on A. longiloba in Guangdong Province, China. Once the disease became severe, plants wilted and died. A fungus was isolated from the diseased stem and identified as Fusarium elaeidis using both morphological characteristics and molecular analysis of DNA-directed RNA polymerase II subunit (rpb2), translation elongation factor-1α (tef1) gene and ß-tubulin (tub2) sequence data. The pathogenicity test showed the fungus was able to produce typical symptoms on A. longiloba similar to those observed in the field. The original pathogen was reisolated from inoculated plants fulfilling Koch's postulates. This is the first report of Fusarium elaeidis causing stem rot on A. longiloba. These results will provide a baseline to identify and control diseases associated with A. longiloba.

Transcriptome analysis of resistant and susceptible mulberry responses to Meloidogyne enterolobii infection.

BACKGROUND: Mulberry (Morus alba L.) is an important sericulture crop; however, root-knot nematode infection seriously limits its production. Understanding the mechanism of interaction between mulberry and nematode is important for control of infection. RESULTS: Using sequencing and de novo transcriptome assembly, we identified 55,894 unigenes from root samples of resistant and susceptible mulberry cultivars at different stages after infection with the nematode Meloidogyne enterolobii; 33,987 of these were annotated in the Nr, SWISS-PROT, KEGG, and KOG databases. Gene ontology and pathway enrichment analyses of differentially expressed genes (DEGs) revealed key genes involved in hormone metabolic processes, plant hormone signal transduction, flavonoid biosynthesis, phenylpropanoid biosynthesis, and peroxisomal and photosynthetic pathways. Analysis of key trends in co-expression networks indicated that expression of unigenes 0,015,083, 0,073,272, 0,004,006, and 0,000,628 was positively correlated with resistance to M. enterolobii. Unigene 0015083 encodes tabersonine 16-O-methyltransferase (16OMT), which is involved in alkaloid biosynthesis. Unigene 0073272 encodes a transcription factor contributing to nitric oxide accumulation during plant immune responses. Unigenes 0,004,006 and 0,000,628 encode ERF and MYB transcription factors, respectively, involved in plant hormone signaling. We verified the accuracy of transcriptome sequencing results by RT-qPCR of 21 DEGs. CONCLUSIONS: The results of this study increase our understanding of the resistance mechanisms and candidate genes involved in mulberry-M. enterolobii interaction. Thus, our data will contribute to the development of effective control measures against this pathogen.

Study of the Compositional, Microbiological, Biochemical, and Volatile Profile of Red-Veined Cheese, an Internal Monascus-Ripened Variety.

Red-Veined Cheese is an experimental internal mold-ripened cheese using red koji powder as the red starter. The objective of this study was to characterize the physicochemical parameters, microorganisms, proteolysis, lipolysis, and volatile profile of the cheese during 33 days of ripening. The gross composition was 56.7% (w/w) for total solids, 45.8% (w/w) for fat-in-dry matter, 2.58% (w/w) for salt content and 37.8% (w/w) for protein. The pH increased from 4.88 to 5.23 during ripening. The Monascus density first rose and then fell, while total mesophilic bacteria count declined steadily throughout the maturation. Proteolysis degree in experimental group was significantly higher than in control group without inoculation of red koji powder. Analysis of the fatty acid profile showed that the internal Monascus-fermentation also promoted lipolysis. A total of 63 volatile compounds, including 12 ketones, 14 alcohols, 15 acids, 13 esters, 5 aldehydes, 3 lactones and 1 phenol, were identified by gas chromatography-mass spectrometry (GC-MS) coupled with headspace solid-phase microextraction (HS-SPME). The main chemical groups of volatile compounds were ketones, alcohols, acids and esters whereas aldehydes, lactones, and phenols represented only minor components. At the end of ripening, the levels of ketones, alcohols, esters were significantly higher in the experimental cheese compared with the control. Our results therefore show that the internal Monascus-fermentation is not only a suitable technology to form red veins in the paste but also an effective method to impact the composition of volatile compounds in cheese, which can thus distinguish it from other internal mold-ripened cheeses and surface-ripened Monascus-fermented cheese.

A one-step electrochemically reduced graphene oxide based sensor for sensitive voltammetric determination of furfural in milk products.

Designing of fast, inexpensive and sensitive furfural determination methods for dairy milk is crucial in analytical and food chemistry. In this study, an electrochemical sensor was developed for the cathodic determination of furfural using a one-step electrochemically reduced graphene oxide (ErGO) modified glassy carbon electrode (GCE). The morphology and chemical constituents of the obtained ErGO/GCE were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and Raman and X-ray photoelectron spectroscopy (XPS). The results showed that the fast and green electrochemical reduction process effectively eliminated the oxygen-containing groups in GO and produced reduced graphene with a high surface area and improved electron transfer kinetics. In addition, the ErGO based sensor displayed excellent responses for furfural in a Na2HPO4-NaH2PO4 solution (pH = 9.18) with a wide linear range from 2 to 2015 µM and a low detection limit of 0.4 µM (S/N = 3). The reduction mechanism of furfural was also discussed. Furthermore, the feasibility of the sensor was confirmed by the determination of furfural in three milk samples which generated acceptable outputs.

Voltammetric Determination of 5-Hydroxymethyl-2-furfural in Processed Cheese Using an Easy-Made and Economic Integrated 3D Graphene-like Electrode.

The concentration of 5-hydroxymethyl-2-furfural (HMF) is an important quality-related index in milk and milk products. Fast, cost-effective and environmentally friendly determination of HMF is of great significance in milk products control. In this study, a three-dimensional (3D) graphene-like surface (3DGrls) was successfully prepared within 5 min by an electrochemical amperometric pretreatment on a pencil graphite electrode (PGE). The fast-obtained 3D graphene-like surface increased the electrode surface area and enhanced the electron transfer capability without the addition of any harmful chemicals. The morphology and chemical composition of the obtained electrode were characterized by scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and electrochemical impedance spectroscopy (EIS). The results found that the electrochemical response to HMF at the prepared 3DGrls/PGE was 34 times higher than that at PGE. The modified electrode showed a good linear response to HMF in a concentration range of 0.35~116 µM with a low limit of detection (LOD) of 0.099 µM. The integrated electrode also exhibited excellent stability and wonderful antifouling property. Furthermore, the 3DGrls/PGE was successfully applied for the determination of HMF in three processed cheese samples with satisfactory results.

Petunidin-Based Anthocyanin Relieves Oxygen Stress in Lactobacillus plantarum ST-III.

Application of probiotics in the food industry has been hampered by their sensitivity to challenging conditions that reduce their vitality in food matrices. A lot of attempts have been made to promote the growth of these probiotics in the aspect of nutrition demands. Among the other adverse conditions, oxygen stress can restrict the growth of probiotics and has not yet been paid enough attention to. In this study, the effect of a petunidin-based anthocyanin (ACN) on the growth of probiotic Lactobacillus plantarum ST-III was investigated under oxygen stress. The growth of ST-III was analyzed through spot assay on agar plates as well as plating-based enumeration of the viable cells in the liquid culture. Results indicated that ACN could efficiently improve the growth of ST-III under oxygen stress, whereas no effect was observed in the absence of oxygen stress. Further investigations indicated that ACN reduced the oxido-reduction potential of the culture; meanwhile, it exerted a positive transcriptional regulation on the thioredoxin system of ST-III, leading to a decrease in reactive oxygen species accumulation within the cells. Moreover, ACN enabled the growth of ST-III in reconstituted skim milk and promoted the formation of milk clots. These results revealed the role of a petunidin-based ACN in oxygen stress relief and highlighted its potential in manufacture and preservation of L. plantarum-based dairy products.

Genetic Diversity of the root-knot nematode Meloidogyne enterolobii in Mulberry Based on the Mitochondrial COI Gene.

This study explores the genetic diversity and structure of Meloidogyne enterolobii in mulberry in China. The COI mitochondrial gene (mtCOI) in M.enterolobii populations in Guangdong, Guangxi, and Hunan Provinces was PCR-amplified, sequenced, and analyzed for genetic diversity. The total number of variations, haplotypes (Hap), the average number of nucleotide differences (k), haplotype diversity (H), and nucleotide diversity (π) of mtCOI were 25, 11, 4.248, 0.900, and 0.00596, respectively. Insignificant differences in Fst value (0.0169) and a high level of gene flow (7.02) were detected among the 19-mulberry root-knot nematode populations, and high genetic variation within each population and a small genetic distance among populations were observed. Both phylogenetic analyses and network mapping of the 11 haplotypes revealed a dispersed distribution pattern of 19 mulberry root-knot nematode populations and an absence of branches strictly corresponding to the 19 range sampling sites. The neutrality test and mismatch analysis indicated that mulberry root-knot nematode populations experienced a population expansion in the past. The analysis of molecular variance (AMOVA) revealed that the genetic differentiation of M. enterolobii was mainly contributed by the variation within each group. No significant correlation was found between the genetic distance and geographical distance of M. enterolobii populations. The findings of this study provide a profound understanding of the M. enterolobii population and will inform the development of strategies to combat and manage root-knot nematodes in mulberry.

Effects of heat treatment, homogenization pressure, and overprocessing on the content of furfural compounds in liquid milk.

BACKGROUND: Sterilization of milk is aimed at killing the microorganisms present. There are three main sterilization methods commonly used in milk processing: high temperature and short time (HTST) pasteurization, ultrahigh pasteurization (UP), and ultrahigh temperature (UHT) sterilization. The Maillard reaction is of special interest in studying the effect of heat treatment on milk quality. Furfural compounds are one of the typical intermediates of the Maillard reaction, which have safety risks related to mutagenic and genotoxic effects. The furfural compounds content is directly related to the heat treatment intensity. RESULTS: The furfural compounds content was measured using high-performance liquid chromatography with ultraviolet detection in 12 min. Then, 13 levels of heat treatment intensity (combinations of temperature and time) and three levels of homogenization pressure were selected to study the change of the furfural compounds content after different processing technologies in a pilot plant. The results show a higher temperature treatment can stimulate more Maillard reaction intermediates, such as hydroxymethylfurfural and furfural. A temperature regression evaluation model and content prediction models of hydroxymethylfurfural and furfural were developed to quantify the relationship between the furfural content and heat treatment with the data from the pilot plant. CONCLUSION: Based on the temperature evaluation model, the heating temperature of three milk products was evaluated. The homogenization pressures had little effect on the furfural content in liquid milk. The emergence of the furyl methyl ketone and methylfurfural can be detected after overprocessing of the liquid milk. © 2020 Society of Chemical Industry.