This study explored an efficient modified Quick, Easy, Cheap, Effective, Rugged and safe (QuEChERS) method combined with liquid chromatography-electrospray ionization with tandem mass spectrometric detection for the analysis of residues of 76 pesticides in brown rice, barley and corn including acidic sulfonylurea herbicides. Formic acid (1%) acid in acetonitrile and dispersive solid phase extractions used for extraction of pesticides and clean-up of the extract respectively. Two fortified spikes at 50 and 200 ng $g^{-1}$ levels were performed for recovery test. Mean recoveries of majority of pesticides at two spike levels ranged from 73.2 to 132.2, 80.9 to 136.8, 66.6 to 143.5 for brown rice, barley and corn respectively with standard error (CV) less than 10%. Good linearity of calibration curves were achieved with $R^2$ > 0.9907 within the observed concentration ranged. The modified method also provided satisfactory results for sulfonylurea herbicides. The method was applied to the determination of residues of target pesticides in real samples. A total of 26 pesticides in 36 out of 98 tasted samples were observed. The highest concentration was observed for tricyclazole at 1.17 mg $kg^{-1}$ in brown rice. This pesticide in two brown rice samples exceeded their MRLs regulated for rice in republic of Korea. Except tricyclazole none of the observed pesticides' concentration was higher than their MRLs. The results reveal that the method is effectively applicable to routine analysis of residues of target pesticides in brown rice, barley and corn.
Single ascospore isolates of Cordyceps bassiana were observed for their colony pigmentation on Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates and were inoculated in a brown rice medium for production of fruiting bodies. Colony pigmentation did not show any relationship with perithecial stromata formation. The isolates were also grown on opposite sides of SDAY agar plates and were observed for vegetative compatibility. Neither vegetative compatibility nor perithecial stromata could be found to be related to each other. It was concluded that fertile fruiting body production was independent of colony pigmentation and vegetative compatibility. Synnemata formation was found to be more common than perithecial stromata formation. This might be due to its highly conidiogenous anamorphic stage, i.e., Beauveria bassiana.
To investigate the constituents of antimutagenic factor from brown rice, methanol extracts were fractionated into ether, ethylacetate, buthanol and aqueous fractions. The ether fractions showed distinct antimutagenic effect and active spot were selected by silica gel chromatography. The specific activity of active spot decreased with isolation of the active components from the methanol extract. Qualitative analyses of the active spot by using various spraying reagents revealed that ninhydrin and orcinol did not develop colored reactions. But, ferric chloride, 2,7-dichlorofluorescein, antimony pentachloride, phosphomolybdic acid, bromothymol blue and rhodamine 6G led to colored reactions. These results suggested that the consitituents of active material were neither polar nor nitrogen-containing compounds and that they may contain phenolic compounds and fatty acid derivatives. Main compounds of the active spot were analyzed to be o-hydroxy benzyl alcohol(saligenin), octanoic acid(caprylic acid), 9,12-cis-octadecadienoic acid(linoleic acid), 11-cis-octadecenoic acid(oleic acid), hexadecanoic acid(palmitic acid), 1H-indole-2-carboxylic acid and 1,2-benzenedicarboxylic acid(phthalate) in GC/Mass spectrum, and antimutagenicity of these active compounds using standard regeant was reconfirmed in S. typhimurium reversion assay.
The primary objective of this study is to determine whether a diet supplemented with brown rice koji (BRK) results in a reduced stress response in rats and mice. BRK, which has been suggested as a candidate for use as a stress- and fatigue-fighting supplement, was compared with red ginseng extract (RG) for its stress-reducing potential. The animals in this study were divided into no-stress, stress, RG, and BRK groups of 8 to 10 animals each. Stress was induced by means of immobilization (being restrained in plastic tubes for 30 min and electroshock (0.5 mA in mice or 2 mA in rats for 5 min). The no-stress group was not exposed to stress. Rats in the RG group received oral doses of 200 mg RG extract/kg body weight daily. The BRK group was fed a 30% BRK diet and exposed to stress. Animals were given supplements for 7 days before being exposed to stress, and then were given supplements for 5 days with exposure to stress. When the stress exposure ended, the animals were observed for stress-related changes in behavior and their plasma corticosterone levels were measured. BRK supplementation was associated with a partial blockade of the effects of stress on locomotion and elevated plus-maze test results in rats and mice. It was also associated with a partial reduction in stress-induced behaviors such as freezing, burrowing, smelling, face-washing, and rearing. BRK supplementation did not have a significant effect on plasma corticosterone levels, which were increased in the animals exposed to stress (p<0.01). The mice in the RG group received RG in water (2 mg RG/ mL $H_2O$), and the BRK group received a 30% BRK diet (weight) for 7 days. Both groups were evaluated for signs of fatigue. BRK supplementation increased endurance, as indicated by time on the rota-rod, in cold water, and on the horizontal wire. These results suggest that BRK supplementation partially protects the animal from the effects of stress and may also contribute to resistance to fatigue on physical exertion.
This study aimed to determine the effect of different dietary level of rice bran extract (RBO) on the laying performance, egg quality, blood parameter, cholesterol, and fatty acids in yolk of Hy-Line Laying hens. In all, 144 Hy-Line Brown laying hens (29 weeks old) were randomly allocated to one of 4 dietary treatments, with 4 replicates per treatment. A commercial basal diet was used and three additional diets were prepared by supplementing 2.5, 5.0 or 10.0 g/kg of RBO to the basal diet. The experimental diets were fed on an ad libitum basis to the bird during 8 weeks. Hen-day egg production increased (quadratic, p<0.05) with inclusion level of RBO, but feed intake, egg weight, and egg mass were not influenced by inclusion of level of RBO in diet. However, the supplementation of RBO did not have an effect on eggshell strength, eggshell thickness, egg yolk color, and HU during the feeding trial. There were no significant differences in the level of leukocyte. However, heterophil and lymphocytes decreased (quadratic, p<0.01) with inclusion level of RBO. H:L ratio tended to decrease (linear p=0.08) with inclusion level of RBO. As expected, increasing inclusion level of RBO in diets decreased (linear, p<0.01) the concentrations of total cholesterol in plasma. AST, ALT, glucose, and albumin were not affected by inclusion of RBO in diets. Egg yolk cholesterol increased (linear and quadratic, p<0.05) with inclusion level of RBO in diet. The results of this study indicate that dietary supplementation of RBO improves laying performance and decreased total cholesterol and egg yolk cholesterol levels in laying hens. Therefore, dietary RBO is considered a valuable functional ingredient to improve the performance of birds.
The chemical media composition and culture conditions were optimized for mycelial growth of Phellinus igniarius 26005. The method of solid-state fermentation, cultivation of basidiomycetal strains in various grains, was developed. Media composition for optimal growth of Phellinus igniarius 26005 was made of 7.0% malt extract, 0.3% bacto soytone, and 0.2% yeast extract. The optimum condition for mycelial growth was $28^{\circ}C$ and pH 7.0, respectively. For the mass cultivation of mycelia, the hydrated grains with cold water, were put into the plastic bottle. The mycelial growth rate in the bottled grains was high in the early stage with inoculation of homogenized mycelium. The activity of mycelium was maintained by adding sterilized water in the middle of cultivation. The glucosamine content which determins the mycelial growth rate in solid material was in the order of job's tears>barley>black soybean>wheat>malt soybean>brown rice>sorghum>glutinous rice.
The natural full-fat rice bran is reported to contain 8.4 to 14.7 wt % Lipids, but the amount and composition of bran depend on the type of rice, quality of paddy, pretreatments to paddy such as parboiling, type of milling system employed, and the degree of polishing. These lipids are usually mixtures of several class fatty acids containing palmitic acid, linolenic acid, linoleic acid, oleic acid, stearic acid, tocopherol, squalene, etc. In this study the oil rich essential fatty acid (EFA) including squalene was extracted from the domestic brown rice bran using supercritical fluid extraction (SFE) and cosolvent induced SFE process, respectively. And the extracts were analyzed with GC-MSD. The extracted amount of rice bran oil was dependent upon the operating pressure and temperature, and the fatty acid composition of oil was varied with the reduced density (${\rho}_{\gamma}$) of supercritical carbon dioxide. About 70~80% of rice bran oil was extracted in 4hrs. The cosolvent induced SFE process shortened the total extraction time, extracted greater amount of oil than SFE process. Especially squalene which was not found in solvent extract phase was identified in SFE and cosolvent induced SFE process.
It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.
This study investigated acetic acid fermentation properties and antioxidant activity of vinegar by addition of lemon grass to develop high quality vinegar by using lemongreass. Traditional brown rice wine contained 5% lemongrass powder and had an alcohol content of 7.2%. The wine was fermented by Acetobacter. sp. RIC-V and made into lemongrass vinegar (LV). The pH and total acidity of the LV were 3.13% and 7.21%, respectively. Fructose was detected whereas glucose, sucrose, and maltose were not detected. Among organic acids, acetic acid was highest at 3658.6 mg%; trace amounts of lactic acid, citric acid, malic acid, tartaric acid, and oxalic aicd were detected. Of the 17 free amino acids, glutamic acid, histidine, alanine, and proline were mainly detected. To conduct total polyphenol content and ABTS radical scavenging activity, 3% and 5% lemongrass powder (P3LV, P5LV) and 1%, 2%, and 3% of lemongrass extract (E1LV, E2LV, E3LV) were added to LV, respectively. Total phenolics increased as the added lemongrass powder and extract increased. Total phenolics were 490.9, 559.4, and $895.7{\mu}g$ gallic acid equivalents/mL in brown rice vinegar, LV, P5LV. ABTS radical scavenging activities were 43.2%, 58.0%, and 91.0% in brown rice vinegar, LV, P5LV, respectively. These results show that lemongrass vinegar has considerable potential as a high quality functional vinegar with antioxidative effects.
Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.
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