Antimicrobial resistance, virulence-associated genes, and pulsed-field gel electrophoresis profiles of Salmonella enterica subsp. enterica serovar Typhimurium isolated from piglets with diarrhea in Korea

Salmonella enterica subsp. enterica serovar Typhimurium was isolated from diarrheic piglets in 2 periods, 2000-2001 (n = 25) and 2005-2006 (n = 17). To compare the characteristics of the isolates collected during the 2 periods, all isolates were tested for antimicrobial resistance, the presence of virulence genes, and pulsed-field gel electrophoresis (PFGE) patterns. All 42 isolates were resistant to at least 1 of the 20 antimicrobials tested, and 39 (93%) were resistant to 2 or more antimicrobials. One isolate was resistant to 12 antimicrobials. Profiles of antimicrobial resistance revealed 20 resistance types. Several isolates were also resistant to quinolones and expanded-spectrum cephalosporins. Ten isolates (24%) were resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (ACSSuT); only one isolate had been isolated in 2000-2001, indicating that this type of resistance has rapidly disseminated. Polymerase chain reaction (PCR) assays revealed that all the isolates carried invA. Among the 25 strains isolated in 2000-2001, all carried the sipA, sopA, sopD, sopE2, and ssaR genes, and 96% carried sopB and sifA. Among the 17 strains isolated in 2005-2006, all carried sifA, and approximately 90% carried sipA, sopA, sopB, sopD, sopE2, and ssaR. However, only 6 (14%) of the 42 isolates carried spvC. By PFGE analysis, all 42 strains were classified into 4 major clusters, basically by collection period. The genetic similarity according to PFGE suggests that the strains isolated from diarrheic piglets of this region within the same period may be closely related.     

Comparison of isothiocyanate metabolite levels and histone deacetylase activity in human subjects consuming broccoli sprouts or broccoli supplement

Increased consumption of cruciferous vegetables such as broccoli may reduce the risk of various cancers. Myrosinase is required to convert dietary glucosinolates from broccoli into bioactive isothiocyanates. We evaluated isothiocyanate excretion profiles in healthy subjects who consumed broccoli sprouts or broccoli supplement (no myrosinase) with equivalent glucosinolate content. Urinary metabolites of two major isothiocyanates, sulforaphane and erucin, were measured by liquid chromatography coupled with tandem mass spectrometry. Peak excretion of sulforaphane and erucin was higher and occurred sooner in subjects who consumed broccoli sprouts as compared to subjects who consumed the supplement. A subject-dependent shift in the ratio of urinary sulforaphane to erucin metabolites was observed in both groups, indicating conversion of sulforaphane to erucin. Lower histone deacetylase activity was observed in the peripheral blood mononuclear cells only in subjects consuming sprouts. Fresh broccoli sprouts differ from broccoli supplements in regards to excretion of isothiocyanates and bioactivity in human subjects.     

Using Seven Types of GM (1, 1) Model to Forecast Hourly Particulate Matter Concentration in Banciao City of Taiwan

In this study, seven types of first-order and one-variable grey differential equation model (abbreviated as GM (1, 1) model) were used to predict hourly particulate matter (PM) including PM₁₀ and PM_2.5 concentrations in Banciao City of Taiwan. Their prediction performance was also compared. The results indicated that the minimum mean absolute percentage error (MAPE), mean squared error (MSE), root mean squared error (RMSE), and maximum correlation coefficient (R) was 14.10%, 25.62, 5.06, and 0.96, respectively, when predicting PM₁₀. When predicting PM_2.5, the minimum MAPE, MSE, RMSE, and maximum R value of 15.24%, 11.57, 3.40, and 0.93, respectively, could be achieved. All statistical values revealed that the predicting performance of GM (1, 1, x ⁽⁰⁾), GM (1, 1, a), and GM (1, 1, b) outperformed other GM (1, 1) models. According to the results, it revealed that GM (1, 1) GM (1, 1) was an efficiently early warning tool for providing PM information to the inhabitants.     

Effect of Goji (Lycium barbarum) on expression of genes related to cell survival

This study investigated the interrelationship between Lycium barbarum (goji) and gene expression in mouse spleen. Oligomicroarray technology was employed to explore the comprehensive response of gene expression and to screen candidate marker genes in the spleens of mice fed a goji suspension. Goji was micronized by media milling and then used to evaluate the effect of size reduction. The average diameter of nano/submicrometer goji was about 100 nm, which exhibited no cytotoxicity to cell lines IEC-6 (rat normal small intestinal cell line) and Caco-2 (human colon adenocarcinoma cell line). It was found that three genes, TNF, Nfkb1, and Bcl-2, were up-regulated and two genes, APAF-1 and caspase-3, were down-regulated by goji. This phenomenon could be helpful for cytoprotection when cells undergo stress or damage that induces the apoptotic pathway. Size reduction into nano/submicrometer scale enhanced bioactivity.     

Survey of polyphenol constituents in grapes and grape-derived products

A rapid and comprehensive qualitative method has been developed to characterize the different classes of polyphenols, such as anthocyanins, flavonols, phenolic acids, and flavanols/proanthocyanidins, in grape products. The detection was achieved by two runs with the same LC gradient in different MS ionization modes and mobile phase modifiers (positive ionization mode and 0.4% trifluoroacetic acid for anthocyanins and flavonols; negative ionization mode and 0.1% formic acid for phenolic acids and flavanols). From an analysis of the MS and UV data and in comparison with the authenticated standards, a total of 53 compounds were identified, including 33 anthocyanins, 12 flavonols, 4 phenolic acids, and 4 flavanols/proanthocyanidins. With the method developed, a survey was then conducted to qualitatively assess the composition of polyphenols among 29 different grape products including original grape, grape juice, grape wine, and grape-derived dietary supplements, and their chemical profiles were systematically compared. This method provided a comprehensive qualitative insight into the composition of polyphenols in grape-derived products.     

Integrated Omics Strategy Reveals Cyclic Lipopeptides Empedopeptins from Massilia sp. YMA4 and Their Biosynthetic Pathway

Empedopeptins—eight amino acid cyclic lipopeptides—are calcium-dependent antibiotics that act against Gram-positive bacteria such as Staphylococcus aureus by inhibiting cell wall biosynthesis. However, to date, the biosynthetic mechanism of the empedopeptins has not been well identified. Through comparative genomics and metabolomics analysis, we identified empedopeptin and its new analogs from a marine bacterium, Massilia sp. YMA4. We then unveiled the empedopeptin biosynthetic gene cluster. The core nonribosomal peptide gene null-mutant strains (ΔempC, ΔempD, and ΔempE) could not produce empedopeptin, while dioxygenase gene null-mutant strains (ΔempA and ΔempB) produced several unique empedopeptin analogs. However, the antibiotic activity of ΔempA and ΔempB was significantly reduced compared with the wild-type, demonstrating that the hydroxylated amino acid residues of empedopeptin and its analogs are important to their antibiotic activity. Furthermore, we found seven bacterial strains that could produce empedopeptin-like cyclic lipopeptides using a genome mining approach. In summary, this study demonstrated that an integrated omics strategy can facilitate the discovery of potential bioactive metabolites from microbial sources without further isolation and purification.     

Micro nutrient toxicity in French marigold

The influence of elevated levels of micronutrients on the growth and flowering of French marigold (Tagetes patula L.) was investigated. Plants were grown with nutrient solution containing 0.25, 0.5, 1, 2, 3, 4, 5, or 6 mM boron (B), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), or zinc (Zn) and toxicity development was monitored. The threshold micronutrient concentrations that induced visible foliar toxicity symptoms were 0.5 mM B, 4 mM Cu, 4 mM Fe, 2 mM Mn, 1 mM Mo, and 5 mM Zn. The dry matter yields during the 5 week experimental period were reduced when micronutrient concentrations exceeded 0.5 mM B, 3 mM Cu, 3 mM Fe, 6 mM Mn, 0.5 mM Mo, and 5 mM Zn in the fertilizer solution. Leaf chlorophyll contents decreased when the nutrient solution concentrations of Cu, Fe, and Mn were greater than 0.5 mM, 3 mM, and 2 mM, respectively. Visual toxicity symptoms of the six micronutrients were characterized.     

Effect of grassland harvesting frequency and N-fertilization on stocks and dynamics of soil organic matter in the temperate climate

ABSTRACT

Management of grassland may affect the dynamics of soil organic carbon (SOC). Objectives were to analyze the effect of different harvesting frequencies and nitrogen fertilization regimes on SOC and total N stocks in a field trial on a sandy loam to loamy sand soil of a grassland site near Kiel (Germany). Additionally, effects on microbial biomass C (C_mic) and ergosterol (as proxy for fungi) contents, water-stable aggregate size-classes and density fractions were studied. In the surface soil (0–10 cm), SOC and total N stocks, amounts of large water-stable macroaggregates (> 2000 µm) and contents of C_mic and ergosterol were significantly higher under a five cut regime. C_mic (r_Spearman = 0.61) and ergosterol contents (r_Spearman = 0.67) were correlated with amounts of large water-stable macroaggregates suggesting that fungi and microbial biomass play an important role in binding of small macroaggregates into large macroaggregates. The free light fraction of SOM showed significantly higher C concentrations under three cut compared to five cut at 30–60 cm, presumably related to the C/N ratio and the decomposability of root litter. This study indicates the importance of cutting frequency on SOC and total N stocks, amounts of large macroaggregates and contents of C_mic and ergosterol.     

Influence of organic matter and inorganic fertilizer on the growth and nitrogen accumulation of corn plants

Effects of different nitrogen (N) sources on the growth and N accumulation of corn plants were studied on plots treated with a compost, a leguminous green manure, and a peat, respectively, associated with a chemical N fertilizer. The experiment included seven treatments with a no‐fertilization check and a conventional chemical fertilizer treatment. Whole corn plants were sampled, and total N was analyzed at 22, 33, 56, 77, and 120 days after seeding (DAS). The results showed that compost with an adequate amount of chemical N fertilizer could reach a high dry matter yield and a high N accumulation, even higher than those of the conventional chemical N fertilizer treatment. With green manure, a considerable amount of N was mineralized and utilized by the corn plants for growth and resulted in a good yield. Neither the peat nor the compost alone could supply enough N for the growth of corn plants. There were no significant effect of treatments on the distribution of dry matter yield and N accumulated in various organs. The crop growth rate of the corn plants of different treatments were significantly different at the vegetative growth stage, however, there were no significant difference during the grain filling period. The apparent N recovery of various treatments were between 0.22 to 0.51 kg N for each kg N applied.     

Effect of rice plant growth on denitrification in rhizosphere soil

The effect of rice plant growth on the loss of basal nitrogen (N) through denitrification in the rhizosphere of subsurface soil was investigated by the ¹⁵N balance method. Labeled ¹⁵N was applied to the deep soil layer to distinguish between the N losses in the surface and subsurface soils. Denitrification in pots with whole plants (Control) was compared with that in pots with plants cut off at the base of the culm (Pcut) to evaluate the effect of plant growth on denitrification. The upward movement of the applied ¹⁵N from the deep soil was negligible. Thus, the amount of unrecovered ¹⁵N was equal to the amount of N lost through denitrification in the rhizosphere of the subsurface soil (20–150 mm soil depth). In the Control treatment, values of redox potential at 50 and 90 mm soil depths were negative throughout the experimental period. Therefore, it was assumed that the redox potential could not have been the limiting factor for the N loss through denitrification in this experiment. The α-naphthylamine-oxidizing activity of roots decreased drastically after the cutting treatment. The estimated amount of de nitrified ¹⁵N in the rhizosphere of the subsurface soil was significantly higher in the Pcut treatment than in the Control one at 30 and 40 d after transplanting (DAT), while it was comparable in the two treatments at 52 and 64 DAT. Since a greater amount of ¹⁵N loss was found to occur when there was no absorption of ¹⁵N by the plants, the absorption of ¹⁵N by plants may have contributed to the suppression of denitrification. The amount of immobilized ¹⁵N in the Control treatment was larger than that of the Pcut treatment throughout the experiment. N immobilization might have also contributed to the suppression of denitrification in the rhizosphere of the subsurface soil.