The initial composition and structure of microbial community determined the yield and quality of Baijiu during the spontaneous fermentation / La composición inicial y la estructura de la comunidad microbiana determinaron el rendimiento y la calidad del Baijiu durante la fermentación espontánea

The microbiota during pit mud fermentation is a crucial factor in Baijiu brewing since it determines the yield and flavor. However, the impact of the microbial community during the initial fermentation stage on Baijiu quality remains uncertain. Herein, high-throughput sequencing was employed to investigate the microbial diversities and distribution during Baijiu fermentation in individual pit mud workshops at both initial and late stages. During the initial fermentation stage, the bacterial community exerted a more pronounced effect on Baijiu quality than the fungal community. And the high-yield pit mud workshop exhibited lower richness and evenness, as well as greater Bray-Curtis dissimilarity during Baijiu fermentation. Lactobacillus was the dominant genus and biomarker in high-yield pit mud, and it constituted the only genus within the bacterial association network during the late fermentation stage. Fungal communities tended to maintain a simple association network with selected core species. Based on the correlation network, Rhizopus and Trichosporon were identified as biomarkers in Baijiu fermentation process. Together, Lactobacillus and Rhizopus could serve as bio-indicators for Baijiu quality during the initial fermentation stage. Therefore, these findings provided novel insights into microbiota interactions during fermentation and the impact of initial microbiota on final Baijiu quality.(AU)

The initial composition and structure of microbial community determined the yield and quality of Baijiu during the spontaneous fermentation.

The microbiota during pit mud fermentation is a crucial factor in Baijiu brewing since it determines the yield and flavor. However, the impact of the microbial community during the initial fermentation stage on Baijiu quality remains uncertain. Herein, high-throughput sequencing was employed to investigate the microbial diversities and distribution during Baijiu fermentation in individual pit mud workshops at both initial and late stages. During the initial fermentation stage, the bacterial community exerted a more pronounced effect on Baijiu quality than the fungal community. And the high-yield pit mud workshop exhibited lower richness and evenness, as well as greater Bray-Curtis dissimilarity during Baijiu fermentation. Lactobacillus was the dominant genus and biomarker in high-yield pit mud, and it constituted the only genus within the bacterial association network during the late fermentation stage. Fungal communities tended to maintain a simple association network with selected core species. Based on the correlation network, Rhizopus and Trichosporon were identified as biomarkers in Baijiu fermentation process. Together, Lactobacillus and Rhizopus could serve as bio-indicators for Baijiu quality during the initial fermentation stage. Therefore, these findings provided novel insights into microbiota interactions during fermentation and the impact of initial microbiota on final Baijiu quality.

Effects of rotation and Bacillus on the changes of continuous cropping soil fungal communities in American ginseng.

The continuous cropping obstacle is the main factor in leading to difficulty in American ginseng replanting. The dormant microbiota in the soil may be the cause of American ginseng disease and eventually caused continuous cropping obstacles, but there are few studies on the dynamic changes of soil microenvironment after American ginseng planting. In this study, we tracked short-term variation in physicochemical properties, enzyme activities, and fungal communities over time-series in soils with continuous cropping obstacle under crop rotation and probiotic Bacillus treatments. Furthermore, we examined the relationships between the important fungal compositions and the soil properties. The results showed that sucrase, cellulase, urease and acid phosphatase activities were significantly increased, while catalase and dehydrogenase were decreased with treatments time. Rotation treatment significantly affected the diversity, dissimilarity degree and species distribution of soil fungal community with continuous cropping obstacle over a short-term. Moreover, beneficial fungal biomarkers such as Cladorrhinum, Oidiodendron, and Mariannaea were accumulated at 48 h under rotation treatments. Almost all fungal biomarkers were negatively correlated with hydrolases and positively correlated with oxidoreductases and acid phosphatase under crop rotation treatments. This study suggested that compared to probiotic Bacillus, crop rotation can significantly affect soil fungal community structure, especially the enrichment of specific potentially beneficial fungal species. Our findings provide a scientific basis for understanding the dynamic changes of fungal communities and soil properties with continuous cropping obstacle of American ginseng in initial stage of soil improvement.

Responses of Soil Rare and Abundant Sub-Communities and Physicochemical Properties after Application of Different Chinese Herb Residue Soil Amendments.

Microbial diversity in the soil is responsive to changes in soil composition. However, the impact of soil amendments on the diversity and structure of rare and abundant sub-communities in agricultural systems is poorly understood. We investigated the effects of different Chinese herb residue (CHR) soil amendments and cropping systems on bacterial rare and abundant sub-communities. Our results showed that the bacterial diversity and structure of these sub-communities in soil had a specific distribution under the application of different soil amendments. The CHR soil amendments with high nitrogen and organic matter additives significantly increased the relative abundance and stability of rare taxa, which increased the structural and functional redundancy of soil bacterial communities. Rare and abundant sub-communities also showed different preferences in terms of bacterial community composition, as the former was enriched with Bacteroidetes while the latter had more Alphaproteobacteria and Betaproteobacteria. All applications of soil amendments significantly improved soil quality of newly created farmlands in whole maize cropping system. Rare sub-communitiy genera Niastella and Ohtaekwangia were enriched during the maize cropping process, and Nitrososphaera was enriched under the application of simple amendment group soil. Thus, Chinese medicine residue soil amendments with appropriate additives could affect soil rare and abundant sub-communities and enhance physicochemical properties. These findings suggest that applying soil composite amendments based on CHR in the field could improve soil microbial diversity, microbial redundancy, and soil fertility for sustainable agriculture on the Loess Plateau.

Effects of American Ginseng Cultivation on Bacterial Community Structure and Responses of Soil Nutrients in Different Ecological Niches.

American ginseng (Panax quinquefolium L.) is a perennial herbaceous plant widely cultivated in China, Korea, the United States, and Japan due to its multifunctional properties. In northwest China, transplanting after 2-3 years has become the main mode of artificial cultivation of American ginseng. However, the effects of the cultivation process on the chemical properties of the soil and bacterial community remain poorly understood. Hence, in the present study, high-throughput sequencing and soil chemical analyses were applied to investigate the differences between bacterial communities and nutrition driver factors in the soil during the cultivation of American ginseng. The responses of soil nutrition in different ecological niches were also determined with the results indicating that the cultivation of American ginseng significantly increased the soluble nutrients in the soil. Moreover, the bacterial diversity fluctuated with cultivation years, and 4-year-old ginseng roots had low bacterial diversity and evenness. In the first two years of cultivation, the bacterial community was more sensitive to soil nutrition compared to the last two years. Proteobacteria, Actinobacteria, Gemmatimonadetes, Acidobacteria, Firmicutes, and Bacteroidetes dominated the bacterial community regardless of the cultivation year and ecological niche. With the increase of cultivation years, the assembly of bacterial communities changed from stochastic to deterministic processes. The high abundance of Sphingobium, Novosphingobium, and Rhizorhabdus enriched in 4-years-old ginseng roots was mainly associated with variations in the available potassium (AK), total phosphorus (TP), total potassium (TK), and organic matter (OM).

Altered intestinal microbiota composition with epilepsy and concomitant diarrhea and potential indicator biomarkers in infants.

Introduction: The diversity and dysregulation of intestinal microbiota is related to the pathology of epilepsy. Gut microbiota plays an important role in epilepsy, and regulating intestinal microbiota through exogenous intervention can alleviate symptoms. However, there are no studies about the effects of epilepsy-related diarrhea on gut microbiota. Methods: The diversity and dysregulation of intestinal microbiota is related to the pathology of epilepsy. Gut microbiota plays an important role in epilepsy, and regulating intestinal microbiota through exogenous intervention can alleviate symptoms. However, there are no studies about the effects of epilepsy-related diarrhea on gut microbiota. To evaluate changes in gut microbiota structure and composition in patients with epilepsy and associated diarrhea, the structure and composition of the fecal microbiota among patients with epilepsy (EP, 13 cases), epilepsy with diarrhea (ED, 13 cases), and probiotic treatments (PT, 13 cases), and healthy controls (CK, seven cases) were investigated and validated by utilizing high-throughput 16S rRNA sequencing. Results: The results showed that the α-diversity indexes indicated that richness and phylogenetic diversity had no significant differences among groups. However, the variation of ß-diversity indicated that the structure and composition of intestinal microbiota were significantly different among the CK, EP, ED, and PT groups (permutational multivariate analysis of variance, p-value = 0.001). Normalized stochasticity ratio and ß-nearest taxon index indicated that stochastic mechanisms exerted increasing influence on community differences with epilepsy and associated diarrhea. ED microbiome alterations include increased Proteobacteria and decreased Actinobacteria and Firmicutes at the phylum level. Bifidobacterium was the core microbe in CK, EP, and PT, whereas it decreased significantly in ED. In contrast, Escherichia/Shigella was the core microbe in CK and ED, whereas it increased significantly in ED (Tukey's multiple comparisons test, adjusted p-value <0.05). The association network in CK has higher complexity and aggregation than in the other groups. The EP network indicated high connectivity density within each community and high sparsity among communities. The bacterial community network of the ED had a more compact local interconnection, which was in contrast to that of PT. The top 7 microbial amplicon sequence variant-based markers that were selected by machine learning to distinguish the groups of epilepsy, probiotic treatments, and healthy infants had stronger discrimination ability. In addition, ASVs_1 (Escherichia/Shigella) and ASVs_3 (Bifidobacterium) had the most importance in the recognition. Discussion: Our research finally showed that infants with epilepsy, epilepsy with diarrhea, and probiotic treatments exhibit substantial alterations of intestinal microbiota structure and composition, and specific intestinal strains are altered according to different clinical phenotypes and can therefore be used as potential biomarkers for disease diagnosis.

Influences of potassium solubilizing bacteria and K-feldspar on enzyme activities and metabolic activities of the bacterial communities in kiwifruit planting soil.

A pot experiment was conducted with kiwifruit planting soil to evaluate the impacts of potassium solubilizing bacteria (KSB) and K-feldspar on the soil nutrient levels, enzyme activities, and microecological environment. The effects were investigated of three inoculation treatments (T1: K-feldspar, T2: KSB, and T3: KSB with K-feldspar) and a non-inoculation treatment (CK) on the enzyme activities and the metabolic activities of the bacterial communities in kiwifruit rhizosphere soil. The results showed that the total nitrogen, available phosphorus, available potassium, and organic matter contents in T3 were 18.19%, 45.22%, 15.06%, and 4.17% higher, respectively, than those in CK at the end of the experiment (90 days). Compared with CK, T3 significantly increased the invertase, urease, acid phosphatase, and polyphenol oxidase activities. T3 had a higher kiwifruit root activity, but there were no significant differences among the four treatments (P > 0.05). T3 significantly altered the bacterial community diversity, increased the utilization of phenolic compounds and polymers, and decreased the utilization of amino acids. Redundancy analysis indicated that soil nutrients (total nitrogen, available phosphorus, and available potassium) and enzyme activities (urease and acid phosphatase) had more important effects on the metabolic activities of the bacterial communities. Co-inoculation enhanced the soil nutrients, enzyme activities, and bacterial community diversity. KSB co-inoculated with K-feldspar has the potential to improve the soil fertility, microbial metabolic activity and plant growth.

Succession of microbial communities and synergetic effects during bioremediation of petroleum hydrocarbon-contaminated soil enhanced by chemical oxidation.

Biotechnologies integrated with chemical techniques are promising in treating the soils contaminated by petroleum hydrocarbons. Experiments by applying the degrading consortium and the modified Fenton (MF) with the chelator sodium citrate simultaneously were carried out to investigate the effects of the MF reagents on the degradation of total petroleum hydrocarbons (TPHs), changes in enzyme activities and the succession of microbial communities at the 0, 20, 100 and 500 mmol/kg hydrogen peroxide concentration levels. The ratio between hydrogen peroxide, ferrous sulfate and sodium citrate in the MF reagents was 100:1:1. The results indicated that the degradation of TPHs conformed to first-order kinetics and MF treatments increased the total removal rates of TPHs (4.73-24.26%) and activated dehydrogenase and polyphenol oxidase activities. A shift in microbial communities from Proteobacteria to Bacteroidetes was observed during the enhanced bioremediation, and the predominant genus shifted from Pseudomonas with an average relative abundance (ARAs) of 76.61% at the beginning to Sphingobacterium with ARAs of 52.06% at the later stage. The MF reagents at the proper level could simplify the relationship among the community populations, alleviate their competition and strengthen their associations, which would optimize the removal efficiency.

WISP1 Predicts Clinical Prognosis and Is Associated With Tumor Purity, Immunocyte Infiltration, and Macrophage M2 Polarization in Pan-Cancer.

Cancer is becoming the leading cause of death and a major public health problem. Although many advanced treatment strategies are currently in use, the general prognosis of cancer patients remains dismal due to the high frequency of recurrence, metastasis. The identification of effective biomarkers is important for predicting survival of cancer patients and improving treatment efficacy. In this study, we comprehensively analyzed WNT1-inducible-signaling pathway protein 1 (WISP1) expression and explored its correlation with prognosis in pan-cancer using tumor IMmune Estimation Resource (TIMER) and Gene Expression Profiling Interactive Analysis 2 (GEPIA2). We also examined correlations between WISP1 and immunocyte infiltration using TIMER. We identified genes co-expressed with WISP1 using the LinkedOmics database and analyzed associated gene ontology using Metascape. Finally, we constructed protein-protein interaction networks and examined correlations between genes co-expressed with WISP1 and immunocyte infiltration in pan-cancer. WISP1 level differed between human pan-cancer tissues and normal tissues, indicating its potential as a prognostic biomarker. WISP1 expression was correlated with tumor purity and immunocyte infiltration, especially monocyte-macrophage trafficking and M2 polarization. Genes co-expressed with WISP1 were mainly associated with extracellular matrix organization, with collagen members COL6A3, COL5A1, and COL8A1 being key genes correlated with macrophage infiltration and M2 polarization in pan-cancer. Conversely, in certain types of cancer with better prognoses, WISP1 was associated with low M2 macrophage infiltration. These results suggest that WISP1 affect clinical prognosis through associations with tumor purity, immune cell infiltration, and macrophage M2 polarization in pan-cancer, with collagen member proteins may serving as effector molecules of WISP1.

Oxaliplatin resistance is enhanced by saracatinib via upregulation Wnt-ABCG1 signaling in hepatocellular carcinoma.

BACKGROUND: Chemo-resistance in hepatocellular carcinoma (HCC) is a major problem, and acquired drug resistance prevents cancer therapies from achieving complete responses. Molecular targeting therapy presents an opportunity to impede tumor through combination or sequential therapy, while the accurate effect is vague. METHODS: The efficacy of combinations between oxaliplatin and anti-cancer molecular targeting drugs was screened. Strangely, the combined chemotherapy with oxaliplatin and saracatinib induced significantly antagonistic effects. Then the antitumor effects of combined treatment with saracatinib and oxaliplatin were confirmed in wide type HCC as well as in saracatinib- and oxaliplatin-resistant HCC. RNA sequencing was used to explore the resistance mechanism, and the roles of ATP-binding cassette transporter G1 (ABCG1) and Wnt signaling in oxaliplatin resistance were confirmed. RESULTS: Chemotherapy with oxaliplatin and saracatinib individually induced strong anti-HCC effects, while combined or sequential treatment of HCC cells with these two drugs exhibited reduced efficacy compared to treatment with the single drugs. And it was saracatinib treatment caused oxaliplatin resistance. RNA sequencing revealed 458 genes that were altered by treatment with saracatinib and oxaliplatin. Of these, the gene encoding ABCG1 and Wnt-associated genes were significantly upregulated. Upregulation of ABCG1 and oxaliplatin resistance were associated with activation of Wnt signaling. Interference with ABCG1 expression or inhibition of Wnt signaling resulted in reversal of the saracatinib-induced oxaliplatin resistance in HCC. CONCLUSIONS: These studies demonstrated that combined or sequential chemotherapy with oxaliplatin and saracatinib reduced antitumor efficacy, and this antagonism was attributed to the activation of Wnt signaling and upregulation of ABCG1 by saracatinib.

Changes in structure and function of bacterial and fungal communities in open composting of Chinese herb residues.

In this study, dynamic changes in bacterial and fungal communities, metabolic characteristics, and trophic modes in Chinese herb residues open composting for 30 days were analyzed by using high-throughput sequencing, PICRUSt, and FUNGuild, respectively. Bacillaceae and Basidiomycota predominated at the early composting stage, while Proteobacteria and Ascomycota became the dominant phyla during the active phase. Aerobic composting had a significant effect on bacterial metabolic characteristics and fungal trophic modes over the composting time. The function of the bacterial communities changed from environmental information processing to metabolism. Fungal communities changed as well, with the pathogenic fungi decreasing and wood saprotrophs increasing. These results indicated that open composting of Chinese herb residues not only influenced microbial community structure but also changed metabolic characteristics and trophic modes, which became the internal dynamics of composting.

Remodeling of hepatic stellate cells orchestrated the stroma-derived oxaliplatin-resistance through CCN3 paracrine in hepatocellular carcinoma.

BACKGROUND: Hepatic stellate cells (HSCs) have a key role in fibrogenesis and in the filtrates of the hepatocellular carcinoma (HCC) stroma, in which they are remodeled and play a critical role in HCC progression. However, the precise role of HSCs trending, infiltration and paracrine in orchestrating the stroma-derived oxaliplatin-resistance in HCC is still vague. METHODS: The chemo-resistant models were established to explore the correlation between HSC cells and the condition of chemoresistance. The HCC clinical samples were collected to confirm this phenomenon. Then, the relationship between secretory CCN3 from oxaliplatin-resistant HCC and the infiltration of HSCs in associated HCC microenvironment was evaluated. Finally, the role and mechanism of HSCs remodeling in the orchestration of oxaliplatin-resistant HCC were explored. RESULTS: The increased infiltration of HSCs and collagen accumulation were found in the microenvironment of oxaliplatin-resistant HCC. The cDNA profiles of the oxaliplatin-resistant HCC was reanalyzed, and CCN3 was one of the significantly increased genes. In HCC clinical samples, the levels of CCN3 and α-SMA are positively correlated, and high expression of CCN3 and α-SMA are positively associated with malignant phenotype and poor prognosis. Then the enhanced abilities of migration and proliferation of HSCs, and elevation of the cytokines paracrine from HSCs relating to HCC malignancy were proved in vitro and in vivo, and which were related to CCN3-ERK signaling pathway activation. CONCLUSIONS: HSCs remodeling are positively related to CCN3 paracrine in hepatocellular carcinoma, which orchestrated the stroma-derived resistance to chemotherapy in HCC.

CCN2-MAPK-Id-1 loop feedback amplification is involved in maintaining stemness in oxaliplatin-resistant hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is the second leading cause of cancer death worldwide. Chemotherapy is an alternative treatment for advanced HCCs, but chemo-resistance prevents cancer therapies from achieving stable and complete responses. Understanding the underlying mechanisms in chemo-resistance is critical to improve the efficacy of HCC. METHODS: The expression levels of Id-1 and CCN2 were detected in large cohorts of HCCs, and functional analyses of Id-1 and CCN2 were performed both in vitro and in vivo. cDNA microarrays were performed to evaluate the alterations of expression profiling of HCC cells with overexpression of CCN2. Finally, the role of downstream signaling of MAPK/Id-1 signaling pathway in oxaliplatin resistance were also explored. RESULTS: The increased expression of Id-1 and CCN2 were closely related to oxaliplatin resistance in HCC. Upregulation of CCN2 and Id-1 was independently associated with shorter survival and increased recurrence in HCC patients, and significantly enhanced oxaliplatin resistance and promoted lung metastasis in vivo, whereas knock-down of their expression significantly reversed the chemo-resistance and inhibited HCC cell stemness. cDNA microarrays and PCR revealed that Id-1 and MAPK pathway were the downstream signaling of CCN2. CCN2 significantly enhanced oxaliplatin resistance by activating the MAPK/Id-1 signaling pathway, and Id-1 could upregulate CCN2 in a positive feedback manner. CONCLUSIONS: CCN2/MAPK/Id-1 loop feedback amplification is involved in oxaliplatin resistance, and the combination of oxaliplatin with inhibitor of CCN2 or MAPK signaling could provide a promising approach to ameliorating oxaliplatin resistance in HCC.

Effects of grassland afforestation on structure and function of soil bacterial and fungal communities.

Grassland afforestation strongly influences the structure and function of soil microorganisms. Yet the mechanisms of how afforestation could simultaneously alter both the soil fungal and bacterial communities and its implications for ecosystem management are poorly understood, especially in nitrogen-limited ecosystems. Using high-throughput sequencing of 16S rRNA and ITS rRNA genes, the present study investigated the changes in soil properties and soil microorganisms after afforestation of natural grasslands with Chinese pine (Pinus tabuliformis) on the Loess Plateau in China. Results showed that soil bacterial diversity had no significant differences among the grassland (GL), forest-grassland transition zone (TZ), and forestland (FL), while soil fungal diversity in the GL was significantly higher than that in the FL and TZ (P < 0.05). The proportion of shared OTUs in the soil bacterial community was higher than that in the soil fungal community among the three land use types. The dominant bacterial phylum shifted from Proteobacteria to Actinobacteria, while the dominant fungal phylum shifted from Ascomycota to Basidiomycota after the GL conversion to the FL. The functional groups of ECM fungi increased significantly while biotrophic fungi decreased significantly after grassland afforestation. Both the soil bacterial and fungal communities in the TZ showed great similarity with those in the FL. In addition, among all examined soil properties, soil nitrogen (N) showed a more significant effect on the soil microbial communities. The reduction of soil N after grassland afforestation resulted in both the structure and function changes in soil microbial communities. Our results demonstrated simultaneously differential changes in the composition and diversity of both soil bacterial and fungal communities after afforestation from grasslands to planted forests.