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Effect of Soybean Meal and Soluble Starch on Biogenic Amine Production and Microbial Diversity Using In vitro Rumen Fermentation

  • Jeong, Chang-Dae;Mamuad, Lovelia L.;Kim, Seon-Ho;Choi, Yeon Jae;Soriano, Alvin P.;Cho, Kwang Keun;Jeon, Che-Ok;Lee, Sung Sil;Lee, Sang-Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.50-57
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    • 2015
  • This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

Distinct Spatio-temporal Expression Patterns of Patatin Promoter-GUS Gene Fusion in Transgenic Potato Microtubers (형질전환 감자 소괴경의 발달단계에 따른 Patatin Promoter-GUS 유전자의 발현 분석)

  • Youm, Jung-Won;Kim, Mi-Sun;Lee, Byoung-Chan;Kang, Won-Jin;Jeon, Jae-Heung;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.13-18
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    • 2003
  • This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

Conversion of Fermented Feed by Basidiomycetes (담자균(擔子菌)을 이용(利用)한 발효(醱酵) 사료(飼料)에 관한 연구(硏究))

  • Hong, Jae-Sik;Kim, Joong-Man;Jeong, Jin-Chul;Lee, Tae-Kyu;Kim, Dong-Han;Kim, Myung-Kon;Lee, Keug-Ro
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.157-168
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    • 1985
  • To investigate nutritive values of a feed fermented with basidiomycetes, among the isolated strains, Lyophyllum decastes (Fr.) Sing. was found with the greatest enzyme productivity and rapid mycelial growth in rice straw medium. Optimum temperature, pH and moisture content for mycelial growth and enzyme production of the strain were $25{\sim}30^{\circ}C,\;pH\;4.0{\sim}7.0\;and\;70{\sim}75\;%$, respectively. Fifteen days of culture were required for the highest enzyme productivity. Among the sub-materials added, $30{\sim}40\;%$ of rice bran and $10{\sim}20\;%$ of defatted perilla seeds were effective for the enzyme production, but caused a reduced mycelial growth. The greatest effect of an addition of inorganic salts was obtained with $0.36{\sim}0.72\;%\;of\;(NH_4)_2HPO_4$. When 40 mesh or smaller rice straw and steam treatment at $0.5\;kg/cm^2$ were used, the mycelial growth decreased, whereas the enzyme production increased. The mycelial growth and enzyme production increased when $Ca(OH)_2$ was used as the alkali treatment, but decreased with increasing concentration of NaOH. As the fermentation proceeded, the amounts of ash, reducing sugar and total nitrogen increased, but cellulose, lignin and pentosan decreased. When the rice straw was treated with alkali, the amounts of ash, total nitrogen and lignin decreased, but reducing sugar and cellulose increased. At higher NaOH concentration, the variation become greater. The in vitro dry matter digestibility of the products increased from 55.03 % at the beginning of the fermentation to 62.72 % at 45 days after fermentation. The most effective alkali treatment on the digestibility of rice straw was KOH followed by NaOH. However, the digestibility increased with increasing concentration of NaOH. The digestibility of pretreated with alkali increased after fermentation as well.

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Laminin Expression in the Rat Lung Development (흰쥐 폐 발생시 Laminin의 발현에 대한 연구)

  • Chung, Ho-Sam;Park, Chul-Hong;Paik, Doo-Jin;Baik, Tae-Kyung;Kim, Won-Kyu;Youn, Jee-Hee;Suh, Yun-Kyung
    • Applied Microscopy
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    • v.31 no.1
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    • pp.71-83
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    • 2001
  • Laminin, a kind of multidomain glycoproteins, is mainly localized in the basement membranes of various tissues. It is known that laminin plays an important part in mammalian lung morphogenesis. The authors have undertaken this study to investigate the changes in the distribution of laminin, and to find out cells which synthesize laminin during the organogenesis and differentiation of the lung. The fetal and neoantal rats (Sprague-Dawley strain) were used as experimental animals. The immunohisto-chemical methods were employed for detection of laminin within the developing lung tissue and the immunegold cytochemical methods were performed for detection of cells which synthesize laminin according to each stage of development. The results are as follows; 1. During fetal life, strong immunoreactivity for laminin is maintained in the basement membranes of the blood vessels and the bronchioles, the extracellular matrix of the mesenchyme, and basal lamina of the alveolar septum in the fetal rat lung. 2. After birth, laminin immunoreactivity at the alveolar septum is gradually reduced. 3. During fetal life, laminin is mainly detected within the cytoplasm of the mesenchymal cells, the endothelial cells of blood vessels and the fibroblasts in fetal rat lung. 4. According to the differentiation of type I and type II pneumocyte after birth, laminin is detected within cytoplasm of the type I pneumocytes, type II pneumocytes and fibroblasts. It is consequently suggested that laminin is largely expressed in the developing lung and laminin may be also synthesized by the type II pneumonocytes at early newborn stages.

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Studies on the Fermentation on Soy Sauce by Bacteria (세균(細菌)을 이용(利用)한 간장 제조(製造)에 관(關)한 연구(硏究))

  • Ju, Hyun-Kyu;Ro, Sin-Kyu;Im, Moo-Hyun
    • Korean Journal of Food Science and Technology
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    • v.4 no.4
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    • pp.276-284
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    • 1972
  • A strain of Bacillus sp. which has powerful enzymatic activity and sets free a little amount of ammonia nitrogen and large amount of amino nitrogen was separated from Meju, Denzang, Chungkukjang, and paddy straw to make the soy-sauce rapidly and conveniently in the various mixing ratio of steamed soy-bean and wheat parched or steamed. Total nitrogen and amino nitrogen were increased during the maturing. The acidity of soy-sauce was increased to $1.15{\sim}1.98%$ than ordinary soy-sauce. pH were ranged in $4.6{\sim}6.0%$. The fermenting time takes 30 days while it takes at least 4 monthes in ordinary method. The best results were obtained with the mixed ratio of 1 : 1 or 1 : 2 (wheat : soy-bean).

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Optimization of a Medium for the Production of Cellulase by Bacillus subtilis NC1 Using Response Surface Methodology (반응 표면 분석법을 사용한 Bacillus subtilis NC1 유래 cellulase 생산 배지 최적화)

  • Yang, Hee-Jong;Park, Chang-Su;Yang, Ho-Yeon;Jeong, Su-Ji;Jeong, Seong-Yeop;Jeong, Do-Youn;Kang, Dae-Ook;Moon, Ja-Young;Choi, Nack-Shick
    • Journal of Life Science
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    • v.25 no.6
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    • pp.680-685
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    • 2015
  • Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R2) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

Effect of Feeding Aspergillus oryzae Inoculant Food-waste Diets on Performance, $NH_3$ Emission and Fecal Microflora in Broiler Chickens (Aspergilius Oryzae 접종 남은 음식물 사료가 육계의 생산성, $NH_3$ 발생량 및 분내 미생물 성상에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.281-289
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    • 2005
  • This study investigated the effect of dried food-waste diets(FW) fermented by Aspergillus oryzae(AO), on broiler growth performance, $NH_3$, emission and fecal microflora. Three hundreds broilers, two week old Hubbard strain, were randomly allotted to 4 experiments and fed with standards early boiler diet replaced with FW and AFW. In experiment 1, eighty four broilers were distributed into 7 treatments with 4 pens at 3 birds per replicate(pen). The dietary treatments ; T1 was com-soy bean meal based broiler diet(Control), T2, T3, T4 were for basal diet replaced with dried food waste without AO(FW) at the level of 20, 40 and $60\%$, respectively and T5, T6 and T7 followed the same levels for the basal diet but using Aspergillus oryzae inoculate food-waste(AFW). For experiments 2, 3, 4, seventy two broilers were distributed into 6 treatments with 4 pens at 3 birds per replicate(pen), respectively. The dietary treatments were the com-soy bean meal based broiler diet replacement with different combinations of FW and AFW, 1:0, 3:1, 1:1, 1:3, 0:1. at level of 20, 40 and $60\%$, respectively. In Exp. 1, it tended to be decreased in weight gain, however, there were no statistical differences among treatments except FW $60\%$ level of replacement(p<0.05). Feed intake and feed efficiency was not different among treatments. Total bacterial counts were not different between the control and FW diet, but E. coli decreased as the AFW levels of replacement were increased(p<0.05). There were no differences in weight gain, feed intake and feed efficiency among treatments in Exp. 2 and weight gains were lower fur FW diet compared with the control and AFW diet in Exp. 3(p<0.05). In Exp. 4, there were no differences in feed intakes among treatments, but lower in weight gain and feed efficiency in FW diet than that the control. In experiment 3, the $NH_3$ emission was the highest among treatments in FW/AFW 1:0 diet(p<0.05). From these results, it seems that FW would be supplemented up to $20\%$ in broiler diets and AO culture extract could improve FW value as feed supplements.

Properties of Organic Acids and Volatile Components in Brown Rice Vinegar Prepared Using Different Yeasts and Fermentation Methods (효모 종류 및 발효 방식에 따른 현미식초의 유기산과 휘발성분 특성)

  • Yoon, Sung-Ran;Kim, Gui-Ran;Lee, Ji-Hyun;Lee, Su-Won;Yeo, Soo-Hwan;Jeong, Yong-Jin;Kwon, Joong-Ho
    • Food Science and Preservation
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    • v.17 no.5
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    • pp.733-740
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    • 2010
  • Brown rice vinegars were prepared by agitated or static acetic acid fermentation using different yeast strains (Saccharomyces kluyveri DJ97, Saccharomyces cerevisiae JK99, Saccharomyces cerevisiae GRJ, or Saccharomyces cerevisiae H9). Organic acid contents and levels of volatile compounds were compared in vinegars prepared by different methods. The chosen yeast strain did not significantly affect the organic acid content of vinegar. In vinegars prepared by agitated acetic acid fermentation, organic acid contents were, in the order of descending abundance, acetic acid, citric acid, lactic acid, oxalic acid, and tartaric acid. In vinegars prepared by static acetic acid fermentation, no citric acid was detected, and lactic acid content was higher than that in agitated acetic acid fermented vinegar. The volatile compounds of both vinegars, analyzed by GC-MS, did not significantly differ when various yeast strains were used. Eighteen volatile compounds were detected in vinegar prepared by agitated acetic acid fermentation and 11 in vinegar prepared by static fermentation. Volatile compounds that can affect vinegar quality, including ethyl acetate and phenethyl acetate, were present at high concentrations in static acetic acid fermented vinegar. Electronic nose analysis showed that volatile chemical patterns differed between the two types of vinegar, but there were no significant differences in sensory scores between vinegars prepared using various yeast strains or by either of the two methods of fermentation.

Malacological Studies on Parafossarulus manchouricus(Gastropoda: Prosobranchia) in Korea (한국산(韓國産) 왜우렁(Parafossarulus manchouricus)의 패류학적(貝類學的) 연구(硏究))

  • Chung, Pyung-Rim
    • The Korean Journal of Malacology
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    • v.1 no.1
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    • pp.24-50
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    • 1985
  • Five different populations of Parafossarulus manchouricus (Chongpyung, Chinju and Kunsan, Korea; and Japan and Taiwan), a population of Bitbynia (Gabbia) misella (Gongju, Korea) and two different populations of Bithynta tentaculata (Michigan, U.S.A. and Bodensee, Germany) were compared in regard to eff-laying characteristics, morphology, chromosome cytology, natural infections of parasites and ecology of habitats. A satisfactory culture method was devised for laboratory rearing of the snails. Tropical fish food (Terra SML) and powdered green leaves (Ceralife) were used as the main food sources for the snails. Benthic diatoms such as Navicula and Gomphonema from the periphyton were also essential for satisfactory growth, especially for the baby snails. The aquaria were stabilized with small stones from a local stream. Young P. manchouricus snails grew to adult size in about 54 days after hatching. They laid eggs 150-156 days after hatching. The whole cycle (birth to egg-laying) took approximately 5 months. The three species of bithyniid snails are iteroparous and lay eggs once a year. There were no major morphological differences in the shells of genera or subgenera studied here. They did exhibit the following rather minor differences. The shell of Parafossarulus has spirally raised ridges, and its apex is usually eroded; the other two genera lack these characteristics. The shell of B. (Gabbia) misella is small, nor exceeding 7.5 mm in length, while the shells of the other two species are larger, being more than 10 mm in length. Scanning electron microscopy (SEM) of the protoconch of P. manchouricus reveals nearly smooth sculpture with small, low, spiral wrinkles. This sculpture is quite different from that of the Hydrobiidae, a family to which the bithyniids are frequently assigned. Scanning electron microscopy of the radulae of the three bithyniid species showed that their radular morphologies are very similar, but there are some small differences, which may be species-specific. There were some statistical differences in shell heights between the Korean and the other populations of P. manchouricus, and between this species and the other two bithyniids as well. The shell differences between the several populations of Korean P. manchouricus may be related to environment. Edtails of the chromosome cycle of these bithyniid snails are similar to those reported for other snails. No specific differences were observed in the chromosome cycle between the various species and populations of snails employed in this study. Reporred for the first time in molluscs are two darkly stained "nucleolar organizers" during pachyterne stages of meiosis. Two different chromosome numbers were observed in the three bithyniid species: n=17 in B. tentaculata and P. manchouricus, and n=18 in B. (G.) misella. no sex chromosomes or supernumerary chromosomes were seen. There were no morphological differences in karyotypes of three Korean strains of P. manchouricus. The infection rates of cercariae of Clonorchis sinensis in Chinju and Kunsan strains of P. manchouricus were 0.14% and 1.25%, respectively. However, Clonorchis cercariae were found in Chongpyung strain of P. manchouriceu and Gongju strain of B. (G.) misella. The habitats of P. manchouricus around Jinyang Lake were relatively clean without any heavy pollution of aquatic microorganisms and organic materials during the period of this study. The levels of dissolved oxygen (D.O.) and biochemical oxygen demand (B.O.D.) of the water specimens sampled from the study areas ranged from 6.0 to 9.6 ppm and from 0.4 to 1.6 ppm, respectively. Eight metalic constituents from the water samples were also assayed, and all metalic ions detercted were remarkably low below the legal criteria. However, calcium ion in the water samples from the habitats of P. manchouricus was considerably higher than others.

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The Acid-resistant Characteristic of Organic Acid Tolerance Mutant of Leuconostoc paramesenteroides (Leuconostoc paramesenteroides 유기산 내성 변이균주의 내산성 특성)

  • Kim, Young-Hwan;Kim, Hee-Zoong;Oh, Kyun-Sik;Kim, Sun-Young;Lee, Si-Kyung;Kang, Sang-Mo
    • Korean Journal of Food Science and Technology
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    • v.40 no.4
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    • pp.424-429
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    • 2008
  • To investigate the acid tolerance characteristics of the acid-resistant mutant, Leuconostoc paramesenteroides P-200, as a kimchi starter, this study examine proton permeability, ATPase activity, glycolysis activity, $Mg^{2+}$ release, and membrane fatty acid composition, and compared the data to that of its wild-type, L. paramesenteroides LP-W. In the proton permeability experiment, the LP-W and P-200 strains' average maximum half-time $(t_{1/2})$ values for pH equilibration through the cell membrane were approximately 5.7 and 9.3 min in 150mM KCl solution, and 4.2 and 8.3 min in 3% NaCl solution, respectively. Their values and pH levels for maximal specific ATPase activity showed that P-200 had greater activity than LPW. And the results of pH-dependent glycolysis activity showed that P-200 had greater activity than LP-W. Furthermore, after 2 hr at pH 4.0, LP-W and P-200 had percent magnesium release values of approximately 12% and 34%, respectively. A comparison of their membrane fatty acid compositions indicated that C18 and cyclo-C19 were the major different fatty acids between the two strains, and their contents of C18 and cyclo-C19 were 2.5% and not detected, respectively, in LP-W, and 6.4% and 11.4%, respectively, in P-200. These results indicate that the P-200 strain has significantly improved acid tolerance as compared to its wild type, LP-W.