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Molecular Analysis of Colonized Bacteria in a Human Newborn Infant Gut

  • Park Hee-Kyung;Shim Sung-Sub;Kim Su-Yung;Park Jae-Hong;Park Su-Eun;Kim Hak-Jung;Kang Byeong-Chul;Kim Cheol-Min
    • Journal of Microbiology
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    • v.43 no.4
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    • pp.345-353
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    • 2005
  • The complex ecosystem of intestinal micro flora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut. To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA iso-lated from feces on the 1st, 3rd, and 6th day. 16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and $50^{\circ}C$ annealing temperature. Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones ($76.7\%$) of all 325 isolated clones were characterized as known species, while other 105 clones ($32.3\%$) were characterized as unknown species. The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days. Thus, set A primers were better suited for this type of molecular ecological analysis. On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present. The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and Streptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present. The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

Estimation of Nutritive Value of Whole Crop Rice Silage and Its Effect on Milk Production Performance by Dairy Cows

  • Islam, M.R.;Ishida, M.;Ando, S.;Nishida, T.;Yoshida, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.10
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    • pp.1383-1389
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    • 2004
  • The nutritive value and utilization of whole crop rice silage (WCRS), Hamasari, at yellow mature stage was determined by three studies. In first study, chemical composition, in vivo digestibility and metabolizable energy (ME) content of WCRS was determined by Holstein steers. WCRS contains 6.23% CP, its digestibility is 48.4% and estimated TDN is 56.4%. Its ME content was 1.91 Mcal/kg DM. Gross energy (GE) retention (% of GE intake) in steers is only 22.7% most of which was lost through feces (44.7% of GE intake). It takes 81 minutes to chew a kg of WCRS by steers. In another study, the effect of Hamasari at yellow mature stage at three stages of lactation (early, mid and late lactation) and two levels of concentrate (40 or 60%) on voluntary intake, ME content and ME intake, milk yield and composition using lactating Holstein dairy cows were investigated. Total intake increased with the concentrate level in early and mid lactation, but was similar irrespective of concentrate level in late lactation. WCRS intake was higher with 40% concentrate level than with 60% concentrate. ME intake by cows increased with the concentrate level and WCRS in early lactating cows with 40% concentrate can support only 90% of the ME requirement. Milk production in accordance with ME intake increased with the increase in concentrate level in early and mid lactating cows but was similar in late lactating cows irrespective of concentrate level. Fat and protein percent of milk in mid and late lactating cows were higher with for 60% concentrate than 40%, but reverse was in early lactating cows. Solids-not-fat was higher with for 60% concentrate than 40% concentrate. Finally in situ degradability of botanical fractions such as leaf, stem, head and whole WCRS, Hamasari at yellow mature stage was incubated from 0 to 96 h in Holstein steers to determine DM and N degradability characteristics of botanical fractions and whole WCRS. Both DM and N solubility, rate of degradation and effective degradability of leaf of silage was lower, but slowly degradable fraction was higher compared to stem and head. Solubility of DM and N of stem was higher than other fractions. The 48 h degradability, effective degradability and rate of degradation of leaf were always lower than stem or head. In conclusion, voluntary intake of silage ranged from 5 to 12 kg/d and was higher with low levels of concentrate, but milk yield was higher with high levels of concentrate. Fat corrected milk yield ranged from 19 to 37 kg per day. For consistency of milk, early lactating cows should not be allowed more than 40% whole crop rice silage in the diet, but late lactating cows may be allowed 60% whole crop rice silage.

Antibacterial Activity of Pharbitin, Isolated from the Seeds of Pharbitis nil, against Various Plant Pathogenic Bacteria

  • Nguyen, Hoa Thi;Yu, Nan Hee;Park, Ae Ran;Park, Hae Woong;Kim, In Seon;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1763-1772
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    • 2017
  • This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibition concentration (MIC) value of $250{\mu}g/ml$. Among the three solvent layers obtained from the methanol extract of P. nil seeds, only the butanol layer displayed the activity with an MIC value of $125{\mu}g/ml$ against Xap. An antibacterial fraction was obtained from P. nil seeds by repeated column chromatography and identified as pharbitin, a crude resin glycoside, by instrumental analysis. The antibacterial activity of pharbitin was tested in vitro against 14 phytopathogenic bacteria, and it was found to inhibit Ralstonia solanacearum and four Xanthomonas species. The minimum inhibitory concentration values against the five bacteria were $125-500{\mu}g/ml$ for the n-butanol layer and $31.25-125{\mu}g/ml$ for pharbitin. In a detached peach leaf assay, it effectively suppressed the development of bacterial leaf spot, with a control value of 87.5% at $500{\mu}g/ml$. In addition, pharbitin strongly reduced the development of bacterial wilt on tomato seedlings by 97.4% at $250{\mu}g/ml$, 7 days after inoculation. These findings suggest that the crude extract of P. nil seeds can be used as an alternative biopesticide for the control of plant diseases caused by R. solanacearum and Xanthomonas spp. This is the first report on the antibacterial activity of pharbitin against phytopathogenic bacteria.

Assessment of DNA Viability in Long term-Stored Buffy Coat Species for the Korean Multicenter Cancer Cohort (한국인 다기관 암 코호트 시료의 DNA 생활성도 평가)

  • Yang, Mi-Hi;Yoo, Ji-Hyun;Kim, Cheong-Sik;Shin, Ae-Sun;Kang, Dae-Hee;Chang, Soung-Hoon;Park, Sue-Kyung;Shin, Hai-Rim;Yoo, Keun-Young
    • Journal of Preventive Medicine and Public Health
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    • v.36 no.4
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    • pp.373-376
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    • 2003
  • Objectives : Peripheral blood-buffy coat fractions (N=14,956) have been stored at $-70^{\circ}C$ in the headquarter of the Korean Multicenter Cancer Cohort (KMCC), since 1993. To study the future molecular etiology of cancers using specimens of the cohort, properly stored specimens are necessary, Therefore, the DNA-viability of the bully coat samples was investigated. Methods : Buffy coat fraction samples were randomly selected from various collection areas and years (N=100). The DNA viability was evaluate from the UV-absorbent ratios at 260/280nm and the PCH for $\beta$-globin was performed with genomic DNA isolated from the buffy coat. Results : PCR products were obtained from 85 and 98% of the C and H area-samples, respectively, using 50 or $100{\mu}l$ of the buffy coat. There were significant differences in the yields of the PCR-amplifications from the C and H areas (p<0.05), which was due to differences in the homogenization of the buffy coat fractions available as aliquots. The PCR-products were obtained from all of the samples (N=7) stored at the C area-local confer, but the other aliquots stored at the headquarter were not PCR-amplified, Therefore, the PCR products in almost all the samples, even including the DNA-degraded samples, were obtained. In addition, an improvement in the DNA isolation, i,e. approx. 1.6 fold, was found after using extra RBC lysis buffer. Conclusions : PCR products for $\beta$-globin were obtained from nearly all of the samples. The regional differences in the PCR amplifications were thought to have originated from the different sample-preparation and homogenization performance. Therefore, the long term-stored buffy coat species at the KMCC can be used for future molecular studies.

The Use of Apple Pomace in Rice Straw Based Diets of Korean Native Goats(Capra hircus)

  • Ahn, J.H.;Jo, I.H.;Lee, J.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1599-1605
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    • 2002
  • In this study, three different experiments were conducted to evaluate the nutritional value of apple pomace produced in southern areas of the Republic of Korea (South Korea). The effects of combining apple pomace in different ratios with commercial concentrates and rice straw in the diets of Korean native goats (Capra hircus) were examined. In experiment I, in situ DM and CP disappearances from nylon bags incubated in the rumen of goats showed that greater amounts of DM and CP were released from apple pomace than those from concentrates at the later stages of incubation, but only after 48 h for DM and CP, respectively. This was reflected in the higher 'b' value of the slowly degradable fraction of the apple pomace compared to the concentrates. Prior to these times the trend was reversed. In experiment II, Korean native goats were fed a diet containing apple pomace with either rice straw or rice straw and concentrates, and the in vivo nutrient digestibilities compared to animals receiving an alfalfa hay. DM digestibility in the animals given apple pomace plus concentrates with rice straw (66.86%) were similar to the goats given alfalfa hay only (69.09%) but significantly greater than for a diet of rice straw plus concentrates. In experiment III, an in vivo study was conducted to investigate the inclusion of 30 to 60% apple pomace pre-mixed with rice straw, rice bran and concentrates on the nutritional value for Korean native goats. Apple pomace mixed diets had higher DM intakes, nutrient digestibility and nitrogen retention than diets without apple pomace, which may have been due to the higher non-structural carbohydrates (NSC) and less ADF and NDF than those in other treatments. Replacement of concentrates with apple pomace in rice straw based diets of Korean native goats fed either separately (experiment II) or by pre-mixing (experiment III) gave satisfactory feed intake, digestibility, pH of ruminal fluid and production of $NH_3$-N and VFA in the rumen of goats. The results of this study infer that apple pomace can be included at levels of up to 60% in the diets of goats without dramatic effect on the animal.

Isolation and Characterization of Dextrans Produced by Leuconostoc sp. strain JYY4 from Fermented Kimchi

  • Gu, Ji-Joong;Ha, Yoo-Jin;Yoo, Sun-Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.4
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    • pp.758-766
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    • 2015
  • Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.

The Physiological and Ecological Comparisons between Warm (Pleuromamma sp.) and Cold Water Copepod Species (Neocalanus plumchrus) in the Northwestern Pacific Ocean Using Lipid Contents and Compositions (북서태평양에서 난수성(Pleuromamma sp.)과 냉수성(Neocalanus plumchrus) 요각류의 지방 함량 및 구성 분석을 통한 생리/생태 비교)

  • Ko, Ah-Ra;Ju, Se-Jong;Lee, Chang-Rae
    • Ocean and Polar Research
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    • v.31 no.1
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    • pp.121-131
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    • 2009
  • In an effort to better understand the physiological and ecological differences between warm and cold water copepod species in Korean waters using lipid contents and compositions, two species of copepods (Pleuromamma sp. as a warm water species and Neocalanus plumchrus as a cold water species) were collected from the Northwest Pacific and East Sea/Sea of Japan, respectively. The cold water species showed two fold higher lipid contents than the warm water species (11% vs. 5% of dry weight). Wax esters, known as one of the major storage lipid classes, were found to be the dominant lipid class (accounting for 64% of total lipids) in the cold water species, whereas, in the warm water species, phospholipids, which are known as membrane components, were the dominant lipid class (accounting for 43% of total lipids),with a trace amount of the storage lipids as a form of triacylglycerols (${\leq}1%$ of total lipids). With regard to the fatty acid compositions, saturated fatty acids (SAFA), especially 16:0 (about 30% of total fatty acids), were most abundant in the warm water species, whereas the polyunsaturated fatty acids (PUFA), particularly eicosapentaenoic acid (EPA : 20:5(n-3)) (${\geq}16%$ of total fatty acids), were most abundant in the cold water species. Among the neutral fraction of lipids, phytol, originating from the side chain of chlorophyll and indicative of active feeding on phytoplankton, was detected only in the warm water species. Significant quantities of fatty alcohols were detected in cold water species, particularly long-chain monounsaturated fatty alcohols (i.e. 20:1(n-9) and 22:1(n-11)), which are well known to abound in cold water herbivorous copepods. However, only trace amounts of short-chain fatty alcohols were detected in the warm water species. Twelve different kinds of sterols were detected in these copepod species, with cholest-5-en-$3{\beta}$-ol (cholesterol) and cholesta-5, 24-dien-$3{\beta}$-ol (desmosterol) dominating in cold and warm water species, respectively. In addition, for the warm water species (Pleuromamma sp.), we assessed the latitudinal gradients of lipid contents and compositions using samples from three different latitudinal regions (Philippine EEZ, Japan EEZ, and the East China Sea). Although no latitudinal gradients of lipid contents were detected, the lipid compositions, particularly dietary fatty acid markers, varied significantly with the latitude. The findings of this study confirm that the distribution of lipid contents and compositions in copepods may not only indicate their nutritional condition and diet history, but may also provide insights into their living strategies under different environmental conditions (i.e., water temperature, food availability).

Effects of Synchronization of Carbohydrate and Protein Supply on Ruminal Fermentation, Nitrogen Metabolism and Microbial Protein Synthesis in Holstein Steers

  • Seo, Ja-Kyeom;Yang, Ji-Young;Kim, Hyun-J.;Upadhaya, Santi Devi;Cho, W.M.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.11
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    • pp.1455-1461
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    • 2010
  • Three rumen-cannulated Holstein steers were fed three diets, each with a different synchrony index (SI) (LS: 0.77, MS: 0.81, and HS: 0.83), in order to examine the effect of diet on rumen fermentation, nitrogen balance, and microbial protein synthesis. Synchrony index was calculated based on the carbohydrate and crude protein fractions of each ingredient and their degradation rates. Feeding the steers diets with different SIs did not influence dry matter, crude protein, NDF, or ADF digestibility. The concentrations of total and individual VFA in the rumens of steers that were fed the two higher-SI diets were higher than in those fed the low-SI diet (p<0.05), but there was no significant difference between the two higher-SI diets. One hour after feeding, steers on the LS diet had lower ruminal pHs than did those fed the MS or HS diets (p<0.05), and animals on the LS diet generally showed higher ruminal $NH_3$-N levels than did animals on the other diets, with the 4-h post-feeding difference being significant (p<0.05). Steers receiving the LS diet excreted more nitrogen (N) in their urine than did those on the two higher-SI diets (p<0.05), and the total N excretion of those on the LS diet was also higher (p<0.05). Microbial N levels calculated from the concentration of urinary purine derivatives were generally higher when the SI was higher, with the highest microbial protein synthesis being produced by steers on the HS diet (p<0.05). In conclusion, in the current study, ingestion of a synchronous diet by Holstein steers improved microbial protein synthesis and VFA production and decreased total N output.

A Study on the Distribution Characteristics of the Small Village Wetlands in Mountainous Rural Area - Case on Geumsan-gun, Chungnam - (산지 읍면지역 소규모 마을습지 분포 특성 연구 - 충남 금산군을 사례로 -)

  • Park, Mi-Ok;Seo, Joo-Young;Yang, Seung-Bin;Koo, Bon-Hak
    • Journal of the Korean Institute of Landscape Architecture
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    • v.47 no.6
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    • pp.37-44
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    • 2019
  • This study was conducted to identify the distribution characteristics of small village wetlands in Geumsan County, an inland mountain zone, and comparing with Seocheon County, a coastal plain area with different ecological environment characteristics. Using Arc-GIS (v10.1) the village wetland code was extracted to derive the possible location of the village wetland, and the final distribution of the village was obtained by performing indoor judging work based on satellite images, aerial photographs, topographical maps, Korea Land Information System (KLIS), land use level, land cover degree (division), and land use status by local surveying and indoor analyzing. Although Geumsan County (576.66km2) is more than 60% larger than Seocheon County (358.04km2), 607 villages in Geumsan County and 570 villages in Seocheon County are capable of making similar levels of 106.5% of wetlands, but only a fraction of those in Seocheon County were found to be 67.6%. The density of the village wetlands was much lower than that of Seocheon County, a coastal plain area, because there were many mountainous areas in Geumsan County, and most of the wetlands temporarily created for water supply were removed during the analysis phase of the Jeongsa Image, so the actual wetlands of the village were judged to be only two-thirds different from those of Seocheon County.

Analysis of Plant Height, Crop Cover, and Biomass of Forage Maize Grown on Reclaimed Land Using Unmanned Aerial Vehicle Technology

  • Dongho, Lee;Seunghwan, Go;Jonghwa, Park
    • Korean Journal of Remote Sensing
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    • v.39 no.1
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    • pp.47-63
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    • 2023
  • Unmanned aerial vehicle (UAV) and sensor technologies are rapidly developing and being usefully utilized for spatial information-based agricultural management and smart agriculture. Until now, there have been many difficulties in obtaining production information in a timely manner for large-scale agriculture on reclaimed land. However, smart agriculture that utilizes sensors, information technology, and UAV technology and can efficiently manage a large amount of farmland with a small number of people is expected to become more common in the near future. In this study, we evaluated the productivity of forage maize grown on reclaimed land using UAV and sensor-based technologies. This study compared the plant height, vegetation cover ratio, fresh biomass, and dry biomass of maize grown on general farmland and reclaimed land in South Korea. A biomass model was constructed based on plant height, cover ratio, and volume-based biomass using UAV-based images and Farm-Map, and related estimates were obtained. The fresh biomass was estimated with a very precise model (R2 =0.97, root mean square error [RMSE]=3.18 t/ha, normalized RMSE [nRMSE]=8.08%). The estimated dry biomass had a coefficient of determination of 0.86, an RMSE of 1.51 t/ha, and an nRMSE of 12.61%. The average plant height distribution for each field lot was about 0.91 m for reclaimed land and about 1.89 m for general farmland, which was analyzed to be a difference of about 48%. The average proportion of the maize fraction in each field lot was approximately 65% in reclaimed land and 94% in general farmland, showing a difference of about 29%. The average fresh biomass of each reclaimed land field lot was 10 t/ha, which was about 36% lower than that of general farmland (28.1 t/ha). The average dry biomass in each field lot was about 4.22 t/ha in reclaimed land and about 8 t/ha in general farmland, with the reclaimed land having approximately 53% of the dry biomass of the general farmland. Based on these results, UAV and sensor-based images confirmed that it is possible to accurately analyze agricultural information and crop growth conditions in a large area. It is expected that the technology and methods used in this study will be useful for implementing field-smart agriculture in large reclaimed areas.