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The aim of this study was to compare fecal microbiome, plasma, fecal and urine metabolomes, and serum biochemistry of adult companion dogs according to body condition scores. Blood, serum/plasma, urine, and fecal samples were collected from 66 clinically healthy, adult companion dogs of either normal weight (NW), overweight (OW), or obese dogs (OB). analyses included fecal microbiome analyses via 16S ribosomal RNA gene amplicon; sequencing, nontargeted plasma, fecal, and urine metabolomics using liquid chromatography/gas chromatography-mass; spectrometry, and serum biochemistry for each dog. Few significant differences in serum biochemistry and fecal microbiome Operational Taxonomic Unit (OTU) were found between weight groups and there was high OTU variation between individual dogs. NW dogs had higher relative abundance of the genus Eubacterium (log-fold change 4.3, adjusted P value = .003) and lower relative abundance of the family Bifidobacteriaceae (log-fold change −3.6, adjusted P value = .02) compared to OB dogs. The microbiome of NW dogs had higher OTU richness compared with OB dogs. Metabolome analysis showed 185 plasma, 37 fecal, and 45 urine metabolites that significantly differed between NW and OW or OB dogs. There were notable significant differences in relative abundance of several plasma phospholipid moieties and fecal volatile fatty acids between weight phenotypes. The combinations of host and gut microbiota and metabolic shifts suggest a pattern that could help detection of early metabolic changes in overweight dogs before the development of obesity related disease. The results of this study support the need for continued investigation into sensitive measures of metabolic aberrancies in overweight dogs.

Beverage quality in the brewing industry is heavily influenced by ingredient properties. The contribution of raw ingredients such as yeast and hops to beer flavor is well understood. However, the influence of barley genotype and/or environment on flavor (the malt 'source') is largely unexplored. Here, a study was performed to determine (i) if there are metabolite differences among six commercial malt sources, (ii) if differences in malt chemistry are reflected in the chemistry of the beer, and (iii) if the differences in the beer chemistry impact sensory attributes of beer, through flavor and flavor stability. Six distinct sources of malts (six varieties from three maltsters) were brewed into six beers using a recipe designed to evaluate differences in flavor. Metabolomics and ionomics was used to characterize chemical variation among the six malts and beers using UHPLC- and HILIC-MS (non-volatile metabolites), HS-SPME/GC–MS (beer volatiles), and ICP-MS (malt metals). These analyses detected a total of 5042 compounds in malt, of which 217 were annotated and included amines, amino acids, fatty acids/lipids/fatty acyls, saccharides/glucosides/sugar acids/sugar alcohols, carboxylic acid derivatives, organic acids, phenolics/benzenoids, purines, pyrimidines/pyridines, terpenes, and organosulfurs. A total of 4568 compounds were detected in beer, of which 246 were annotated and included esters, aldehydes, and alcohols. Statistical analysis revealed chemical variation among the six malts (50/217 malt metabolites varied) and beers (150/246). The six beers were evaluated for flavor using a modified descriptive analysis for 45 sensory traits at 0, 4, and 8 weeks of storage at 4 °C. Principal component analysis of the sensory data revealed flavor differences among the six beers at 8 weeks, and the malt-type Full Pint was described as fruity and Meredith as corn chip. The metabolite and sensory data were integrated and revealed associations between flavor profiles in beer and the annotated malt and beer. The fruity or corn chip flavor profiles in beer were associated beer purines/pyrimidines, volatile ketones, amines, and phenolics, and malt lipids, saccharides, phenols, amines, and alkaloids. Taken together, these data support a role of malt source in beer flavor and flavor stability. As a raw ingredient, malting barley genotypes can be evaluated for a contribution to flavor, and this may be a future target for plant breeding, agronomy, and malting efforts to selectively improve flavor, flavor stability, and quality in beer.

Limited knowledge of optimal microbial community composition for PPCP biotreatment, and of the microbial phylotypes that drive biotransformation within mixed microbial communities, has hindered the rational design and operation of effective and reliable biological PPCP treatment technologies. Herein, bacterial community composition was investigated as an isolated variable within batch biofilm reactors via comparison of PPCP removals for three distinct inocula. Inocula pre-acclimated to model PPCPs were derived from activated sludge (AS), ditch sediment historically-impacted by wastewater treatment plant effluent (Sd), and material from laboratory-scale soil aquifer treatment (SAT) columns. PPCP removals were found to be substantially higher for AS- and Sd-derived inocula compared to the SAT-derived inocula despite comparable biomass. Removal patterns differed among the 6 model compounds examined (diclofenac, 5-fluorouracil, gabapentin, gemfibrozil, ibuprofen, and triclosan) indicating differences in biotransformation mechanisms. Sphingomonas, Beijerinckia, Methylophilus, and unknown Cytophagaceae were linked with successful PPCP biodegradation via next-generation sequencing of 16S rRNA genes over time. Results indicate the criticality of applying engineering approaches to control bacterial community compositions in biotreatment systems.

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV) is an economically important disease of sugar beet. Management includes the use of single resistance (Rz) genes. However, strains with the ability to overcome the widely used Rz1 resistance gene have already been reported. The mechanisms associated with these genes are unknown, therefore increased understanding of the host defense response may lead to more durable sources of rhizomania resistance. Non-targeted metabolome profiling, coupled with targeted phytohormone quantification, was applied to characterize the metabolic and signaling pathways associated with BNYVV infections in sugar beet. Clear differences in metabolites were identified following pathogen infection independent of plant genotype particularly those associated with host lipid composition or other membrane associated functions.

Rhizoctonia crown and root rot (RCRR), caused by Rhizoctonia solani Kuhn AG 2-2 IIIB, is an important disease of sugar beet. While RCRR can be managed by agronomic practices, plant resistance remains the primary method for control. However, the molecular processes that mediate resistance to R. solani are largely unknown. The metabolic changes that occurred during susceptible and resistant R. solani interactions were compared and characterized using nontargeted metabolomic profiling. Metabolites from infected and healthy, root and leaf tissue, were taken at 0 and 7 dai and detected using reversed-phase UHPLC-MS and GC-MS. There was a clear distinction in the metabolome between tissue type and genotype, and in response to R. solani. 143 compounds were annotated and several metabolites associated with plant defense to fungi were identified in both germplasm.

Background: Dichlorodiphenyldichloroethylene (p,p′-DDE) and hexachlorobenzene (HCB) are organochlorine pesticides with well-known endocrine disrupting properties. Exposure to p,p′-DDE and HCB concerns human populations worldwide and has been linked to metabolic disorders such as obesity and type 2 diabetes, but details about these associations in humans from the general population are largely unknown. Objectives: We investigated the associations between p,p′-DDE and HCB exposure and global metabolomic profiles in serum samples from 1016 participants from the Swedish population-based Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study. Methods: HCB and p,p′-DDE levels were determined using gas chromatography coupled to high-resolution mass spectrometry (GC–HRMS). Metabolite levels were determined by using a non-targeted metabolomics approach with ultra-performance liquid chromatography coupled to time-of- flight mass spectrometry (UPLC–TOFMS). Association analyses were performed using multivariate linear regression. Results: We found circulating levels of p,p-DDE and HCB to be significantly associated with circulating levels of 16 metabolites following adjustment for age, sex, education level, exercise habits, smoking, energy intake, and alcohol intake. The majority of the 16 metabolites belong to lipid metabolism pathways and include fatty acids, glycerophospholipids, sphingolipids, and glycerolipids. Overall, p,p′-DDE and HCB levels were found to be correlated to different metabolites, which suggests that different metabolic fingerprints may be related to circulating levels of these two pesticides. Conclusions: Our findings establish a link between human exposure to organochlorine pesticides and metabolites of key metabolic processes mainly related to human lipid metabolism. Keywords: Organochlorine pesticides, Human exposure, Metabolomics, Metabolic effects, DDT, HCB

To search for novel pathophysiological pathways related to ischemic stroke using a metabolomics approach.We identified 204 metabolites in plasma by liquid chromatography mass spectrometry in 3 independent population-based samples (TwinGene, Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) and Uppsala Longitudinal Study of Adult Men). TwinGene was used for discovery and the other 2 samples were meta-analyzed as replication. In PIVUS, traditional cardiovascular (CV) risk factors, multiple markers of subclinical CV disease, markers of coagulation/fibrinolysis were measured and analyzed in relation to top metabolites.In TwinGene (177 incident cases, median follow-up 4.3 years), levels of 28 metabolites were associated with incident ischemic stroke at a false discover rate (FDR) of 5%. In the replication (together 194 incident cases, follow-up 10 and 12 years, respectively), only sphingomyelin (32:1) was significantly associated (HR .69 per SD change, 95% CI .57-0.83, P value = .00014; FDR5%) when adjusted for systolic blood pressure, diabetes, smoking, low density lipoportein (LDL)- and high density lipoprotein (HDL), body mass index (BMI) and atrial fibrillation. In PIVUS, sphingomyelin (32:1) levels were significantly related to both LDL- and HDL-cholesterol in a positive fashion, and to serum triglycerides, BMI and diabetes in a negative fashion. Furthermore, sphingomyelin (32:1) levels were related to vasodilation in the forearm resistance vessels, and inversely to leukocyte count (P.0069 and .0026, respectively).An inverse relationship between sphingomyelin (32:1) and incident ischemic stroke was identified, replicated, and characterized. A possible protective role for sphingomyelins in stroke development has to be further investigated in additional experimental and clinical studies.

1,2,4-butanetriol (butanetriol) is a useful precursor for the synthesis of the energetic material butanetriol trinitrate and several pharmaceutical compounds. Bacterial synthesis of butanetriol from xylose or arabinose takes place in a pathway that requires four enzymes. To produce butanetriol in plants by expressing bacterial enzymes, we cloned native bacterial or codon optimized synthetic genes under different promoters into a binary vector and stably transformed Arabidopsis plants. Transgenic lines expressing introduced genes were analyzed for the production of butanetriol using gas chromatography coupled to mass spectrometry (GC-MS). Soil-grown transgenic plants expressing these genes produced up to 20 µg/g of butanetriol. To test if an exogenous supply of pentose sugar precursors would enhance the butanetriol level, transgenic plants were grown in a medium supplemented with either xylose or arabinose and the amount of butanetriol was quantified. Plants expressing synthetic genes in the arabinose pathway showed up to a forty-fold increase in butanetriol levels after arabinose was added to the medium. Transgenic plants expressing either bacterial or synthetic xylose pathways, or the arabinose pathway showed toxicity symptoms when xylose or arabinose was added to the medium, suggesting that a by-product in the pathway or butanetriol affected plant growth. Furthermore, the metabolite profile of plants expressing arabinose and xylose pathways was altered. Our results demonstrate that bacterial pathways that produce butanetriol can be engineered into plants to produce this chemical. This proof-of-concept study for phytoproduction of butanetriol paves the way to further manipulate metabolic pathways in plants to enhance the level of butanetriol production.

Fuzhuan tea is a traditional preparation of Camellia sinensis L. (Theaceae) from Hunan, China, that is fermented with the fungus Eurotium cristatum. Metabolomic analysis was performed on fuzhuan tea extracts and compared to extracts of non-fermented green teas using ultra-performance liquid chromatography/time of flight-mass spectrometry (UPLC-ToF-MS). Principal component analysis revealed a unique phytochemical profile between the two types of tea with the largest separation visible along the third principle component, which accounted for 12.4% of dataset variation. Spectral comparison of significantly different tea metabolite features allowed tentative identification of flavonoids including catechins, fatty acid amides, and other lipids and polysaccharides. Fuzhuan tea extracts, at a concentration of 5 mg/mL or less, reduced the growth of enteric pathogens Shigella sonnei and Staphylococcus aureus by 50%, and had a minimum inhibitory concentration (MIC) of 0.625 mg/mL against S. aureus. These results support a distinct phytochemical profile associated with fermented fuzhuan tea compared to non-fermented green teas that warrant further investigation for novel compounds with antimicrobial bioactivity.

The effect of temperature on non-volatile compounds in beer has not been well characterised during storage. Here, a metabolomics approach was applied to characterise the effect of storage temperature on non-volatile metabolite variation after 16weeks of storage, using fresh beer as a control. The metabolite profile of room temperature stored (RT) and cold temperature stored (CT) beer differed significantly from fresh, with the most substantial variation observed between RT and fresh beer. Metabolites that changed during storage included prenylated flavonoids, purines, and peptides, and all showed reduced quantitative variation under the CT storage conditions. Corresponding sensory panel observations indicated significant beer oxidation after 12 and 16weeks of storage, with higher values reported for RT samples. These data support that temperature affected beer oxidation during short-term storage, and reveal 5-methylthioadenosine (5-MTA) as a candidate non-volatile metabolite marker for beer oxidation and staling.

Root-secreted phytochemicals mediate multiple interactions in the rhizosphere. The root exudation process can be altered by various biotic factors, including pathogenic and non-pathogenic microbes, and abiotic factors like temperature and soil moisture. It has been suggested that root secretion of specific flavonoids is influenced by diurnal rhythms (by light or dark) but a comprehensive analysis of the overall secretion of phytochemicals in response to diurnal rhythms has not been studied. In this study, we analyzed the effect of light/dark cycles on root exudation profiles using Arabidopsis as a model plant. Our results reveal that the root secretion of phytochemicals is partly regulated by the diurnal light cycle and follows two main patterns of secretion: (1) the large majority of phytochemicals in the exudates showed no diurnal pattern in their secretion, and (2) a few compounds showed a diurnal pattern in their secretion: three compounds increased in secretion only under light; two compounds increased in secretion only while it was dark; and two compounds increased in secretion during the transition from dark to light. Root-specific ABC transporters have been implicated in root exudation; an analysis of the gene expression patterns of ABC transporters in the roots of Arabidopsis at specific time points revealed that none of the ABC transporters followed a diurnal expression pattern, suggesting that they are expressed constantly during the day and night. Similarly, we analyzed the expression in roots of genes involved in secondary metabolite biosynthesis and found that some of the genes involved in phenylpropanoid and glucosinolate biosynthesis (i.e. 4-coumarate-CoA ligases (4CL1 and 4CL2), flavonol synthases (FS1 and FS2), and CYP79B3) followed distinct diurnal expression patterns. Overall, we have discovered that while root exudation of the majority of phytochemicals is constitutive, the secretion of a few compounds follows a diurnal rhythm, which is in accordance with the expression of some genes involved in secondary metabolism.

Centaurea maculosa is a plant species native to Eurasia and invasive in many regions of North America that secretes a phytotoxin from its roots, (+/−)-catechin, which is thought to serve an allelopathic role by reducing fitness of neighboring species. However, catechin is a relatively unstable compound, is often found at relatively low concentrations, interacts with soil cations to form insoluble complexes and/or degradation products, and exhibits extremely variable accumulation patterns in the soil. These factors, coupled with a lack of knowledge of the regulation of catechin production by C. maculosa , confound our understanding of the importance of catechin as an allelochemical. The time and cost of current sampling procedures effectively limit the observations made on catechin accumulation patterns, without which it is extremely difficult to conclusively attribute allelopathic significance to this compound. Thus, a highly sensitive, fast, inexpensive and reliable method for soil catechin analysis is needed. We report a novel method that utilizes the colorimetric reagent dimethylaminocinnamaldehyde (DMACA) in an acidic ethanol solution for detection of soil catechin. This method is selective and extremely sensitive and can be used in the field for qualitative, but not quantitative, analysis. This assay will allow for a greater understanding of the role of catechin as an allelochemical.

Polyacetylene 3 was said to be the methyl ether of a known alcohol (15 in Bohlmann and Zdero (1970)), but was instead characterized spectroscopically as another isomer. Reassignment of one HMBC correlation now establishes the data and structure into conformity with the Bohlmann/Zdero formulation, depicted below. This also fits 3 into a biosynthetic pattern represented by the other four polyacetylenes isolated. We thank Professor V. Wray for pointing out the discrepancies and for helpful comments. F. Bohlmann and C. Zdero, Uber die inhaltsstoffe aus Eclipta erecta L. Chem Ber. 103 (1970), pp. 834–841. The correct structure, name and NMR spectroscopic assignments for 3 are now given below.