Yangfang Ye

Systems Responses of Rats to Aflatoxin B1 Exposure Revealed with Metabonomic Changes in Multiple Biological Matrices

Exposure to aflatoxins causes liver fibrosis and hepatocellular carcinoma posing a significant health risk for human populations and livestock. To understand the mammalian systems responses to aflatoxin-B1 (AFB1) exposure, we analyzed the AFB1-induced metabonomic changes in multiple biological matrices (plasma, urine, and liver) of rats using (1)H NMR spectroscopy together with clinical biochemistry and histopathologic assessments. We found that AFB1 exposure caused significant elevation of glucose, amino acids, and choline metabolites (choline, phosphocholine, and glycerophosphocholine) in plasma but reduction of plasma lipids. AFB1 also induced elevation of liver lipids, amino acids (tyrosine, histidine, phenylalanine, leucine, isoleucine, and valine), choline, and nucleic acid metabolites (inosine, adenosine, and uridine) together with reduction of hepatic glycogen and glucose. AFB1 further caused decreases in urinary TCA cycle intermediates (2-oxoglutarate and citrate) and elevation of gut microbiota cometabolites (phenylacetylglycine and hippurate). These indicated that AFB1 exposure caused hepatic steatosis accompanied with widespread metabolic changes including lipid and cell membrane metabolisms, protein biosynthesis, glycolysis, TCA cycle, and gut microbiota functions. This implied that AFB1 exposure probably caused oxidative-stress-mediated impairments of mitochondria functions. These findings provide an overview of biochemical consequences of AFB1 exposure and comprehensive insights into the metabolic aspects of AFB1-induced hepatotoxicity in rats.

Global Metabolomic Responses of Escherichia coli to Heat Stress

Microbial metabolomic analysis is essential for understanding responses of microorganisms to heat stress. To understand the comprehensive metabolic responses of Escherichia coli to continuous heat stress, we characterized the metabolomic variations induced by heat stress using NMR spectroscopy in combination with multivariate data analysis. We detected 15 amino acids, 10 nucleotides, 9 aliphatic organic acids, 7 amines, glucose and its derivative glucosylglyceric acid, and methanol in the E. coli extracts. Glucosylglyceric acid was reported for the first time in E. coli. We found that heat stress was an important factor influencing the metabolic state and growth process, mainly via suppressing energy associated metabolism, reducing nucleotide biosynthesis, altering amino acid metabolism and promoting osmotic regulation. Moreover, metabolic perturbation was aggravated during heat stress. However, a sign of recovery to control levels was observed after the removal of heat stress. These findings enhanced our understanding of the metabolic responses of E. coli to heat stress and demonstrated the effectiveness of the NMR-based metabolomics approach to study such a complex system.

Metabolic Phenotypes Associated with High-Temperature Tolerance of Porphyra haitanensis Strains

Colored mutants of Porphyra haitanensis have superior production and quality characteristics, with two mutants, Shengfu 1 (SF-1) and Shengfu 2 (SF-2), having good high-temperature tolerances. To understand the molecular aspects of high-temperature tolerance, this study comprehensively investigated the metabolic differences between the high-temperature tolerant strains and wild type. Nuclear magnetic resonance (NMR) methods identified 35 algal metabolites, including sugars, amino acids, carboxylic acids, aldehydes, amines, and nucleotides. The results indicated that the high-temperature tolerant strains had significantly different metabolic phenotypes from the wild type. The high-temperature tolerant mutants had significantly higher levels in a set of osmolytes consisting of betaine, taurine, laminitol, and isofloridoside than the wild type, indicating the particular importance of efficient osmoregulation for high-temperature resistance. These findings provided essential metabolic information about high-temperature adaptation for P. haitanensis and demonstrated NMR-based metabolomics as a useful tool for understanding the metabolic features related to resistance to stressors.

Variations of Bacterial Community Structure in Flooded Paddy Soil Contaminated with Herbicide Quinclorac

The denaturing gradient gel electrophoresis (DGGE) method was applied to determine the relative genetic complexity of microbial communities in flooded paddy soil treated with herbicide quinclorac (3,7-dichloro-8-quinoline-carboylic acid). The results obtained showed a significant effect of quinclorac on the development of bacterial populations in soils contaminated with different concentrations of the herbicide at the early time after application. In general, however, the number of populations of the same soil sample treated with the same concentration of the quinclorac differed obviously with increasing incubation time within the early 8 weeks. The scale of differences in banding patterns-showed that the microbial community structures of the quinclorac-treated and non-quinclorac-treated soils were not significantly different after 21 weeks of incubation. Quantification, as demonstrated in this paper, was studied by establishing dose-response relationships. Significant pattern variations were quantified. Prominent DGGE bands were excised, cloned and sequenced to gain insight into the identities of predominant bacterial populations. The majority of DGGE band sequences were related to bacterial genera Clostridium, Sphingobacterium, Xanthomonas and Rhodococcus.

Evolution of metabolomics profile of crab paste during fermentation

Crab paste is regularly consumed by people in the coastal area of China. The fermentation time plays a key role on the quality of crab paste. Here, we investigated the dynamic evolution of metabolite profile of crab paste during fermentation by combined use of NMR spectroscopy and multivariate data analysis. Our results showed that crab paste quality was significantly affected by fermentation. The quality change was manifested in the decline of lactate, betaine, taurine, trimethylamine-N-oxide, trigonelline, inosine, adenosine diphosphate, and 2-pyridinemethanol, and in the fluctuation of a range of amino acids as well as in the accumulation of glutamate, sucrose, formate, acetate, trimethylamine, and hypoxanthine. Trimethylamine production and its increased level with fermentation could be considered as a freshness index of crab paste. These results contribute to quality assessment of crab paste and confirm the metabolomics technique as a useful tool to provide important information on the crab paste quality.

Dynamic metabolomic responses of Escherichia coli to nicotine stress

Previously, we reported the metabolic responses of Pseudomonas sp. strain HF-1, a nicotine-degrading bacterium, to nicotine stress. However, the metabolic effects of nicotine on non-nicotine-degrading bacteria that dominate the environment are still unclear. Here, we have used nuclear magnetic resonance based metabolomics in combination with multivariate data analysis methods to comprehensively analyze the metabolic changes in nicotine-treated Escherichia coli. Our results showed that nicotine caused the changes of energy-related metabolism that we believe are due to enhanced glycolysis and mixed acid fermentation as well as inhibited tricarboxylic acid cycle activity. Furthermore, nicotine resulted in the alteration of choline metabolism with a decreased synthesis of betaine but an increased production of dimethylamine. Moreover, nicotine caused a decrease in amino acid concentration and an alteration of nucleotide synthesis. We hypothesize that these changes caused the decrease in bacterial cell density observed in the experiment. These findings provide a comprehensive insight into the metabolic response of E. coli to nicotine stress. Our study highlights the value of metabolomics in elucidating the metabolic mechanisms of nicotine action.

Effect of high-pressure treatment on taste and metabolite profiles of ducks with two different vinasse-curing processes

The effect of high-pressure (HP) (0.1, 150 and 300MPa, 15min) on taste profiles of vinasse-cured ducks was investigated; the metabolite profiles were determined using 1H NMR. HP at 150MPa increased the taste intensity of products compared with the controls, while HP at 300MPa did not further improve their taste compared with 150MPa treated samples. The metabonome of vinasse-cured ducks was dominated by 27 metabolites. HP increased amino acids, glucose, alkaloids and organic acids, but decreased inosine monophosphate and its derivatives, compared with the controls. The increments of metabolites in vinasse-dry-cured duck were higher than those in vinasse-wet-cured duck. The change of metabolites could be related to the enzyme activity, the degradations of proteins, sugars and nucleotides, and the permeation from vinasse-curing agents to duck meat. These findings suggest that 150MPa treatment was effective to improve the taste of vinasse-cured duck.

1H NMR and multivariate data analysis of the differences of metabolites in five types of dry-cured hams

In order to distinguish the taste styles of dry-cured hams (Jinhua, Xuanwei, Country, Parma and Bama), we established a 1H nuclear magnetic resonance spectroscopy method to identify metabolites. Totally, 33 charged metabolites, including amino acids, organic acids, nucleic acids and their derivatives, sugars, alkaloids and others were identified. The abundant glutamate, lysine, alanine, leucine and lactate could be the major contributors of taste. Total variables were explained by PC1 (67.7%) and PC2 (16.0%) which showed that Parma and Xuanwei styles were close to each other (similar amino acids, peptide, organic acids and alkaloids contents). Bama style showed the highest PC1 and amino acids, organic acids and alkaloids contents. Country style was located on the left-most area of PC1 (the lowest amino acids, organic acids and peptide, but the highest sugars contents). Sensory evaluation revealed that Bama ham had the highest overall taste score, followed by Jinhua, Parma, Xuanwei and American Country ham. We concluded that the proportions and combinations of taste components explained the specific taste instead of any single component. These findings provided a better understanding of different metabolomics among hams.