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Effects of Electric Stimulation and Activation Conditions on the Fusion and Development of Porcine Somatic Cell Nuclear Transfer Embryos (전기적 융합과 활성화 방법이 돼지 체세포 복제수정란의 체외발달에 미치는 영향)

  • 정기화
    • Journal of Embryo Transfer
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    • v.19 no.1
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    • pp.43-51
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    • 2004
  • The present study was conducted to investigate the effects of fusion and/or activation protocol on in vitro development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine fetal fibroblast cells were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion and activation were induced simultaneous fusion/activation (SA) or delayed activation (DA) with or without cytochalasin B (CB) treatment with electic pulses in 0.28 M mannitol-based medium. The SCNT embryos were cultured in vitro for 7 days and stained with Hoechst 33342 to determine the number of nuclei. After 7 days culture, cleavage and blastocyst formation rates were 72.4% and 7.6% in SCNT and 76.3% and 20.4% in parthenotes. To examine the effect of electric field strengths on development of SCNT embryos, oocytes were fused two pulses of 110 V/mm, 130 V/mm or 150 V/mm for 30 sec post-injection. The fusion and cleavage rates in 130 V/mm group (70.2% and 72.6%) and 150 V/mm group (72.6% and 70.5%) were higher (P<0.05) than 110 V/mm group (47.1% and 48.6%), respectively. However, the rate of embryos developing to the blastocyst stage (8.1%, 9.7% and 10.7%) were not different among three groups. The cleavage rates and the blastcyst formation rates were not different among three treatment groups (SA group, 71.4% and 9.7%; SA+CB treatment group, 74.7% and 8.0%; DA+CB treatment group, 70.8% and 11.2%, respectively). And, no different in the number of cells in blastocysts was observed among the three groups (22.5$\pm$12.8, 23.3$\pm$11.2 and 21.6$\pm$10.4, respectively). These result suggest that two pulses of 130 V/mm or 150 V/mm for 30 sec with SA treatment or DA treatment are enough for fusion/activation of porcine somatic cell nuclear transfer (SCNT) embryos to develop to the blastocyst stage.

Biochemical Properties of Lactate Dehydrogenase Eye-Specific C4 Isozyme: Lepomis macrochirus and Micropterus salmoides (젖산탈수소효소 eye-specific C4 동위효소의 생화학적 특성: 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides))

  • Yum, Jung-Joo;Ku, Bo-Ra
    • Journal of Life Science
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    • v.22 no.2
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    • pp.209-219
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    • 2012
  • The properties of lactate dehydrogenase (LDH, EC 1.1.1.27) eye-specific $C_4$ isozyme were studied by polyacrylamide gel electrophoresis, Western blotting, immunoprecipitation, and enzyme kinetics. Furthermore, we proposed the optimal conditions for measuring the activity of LDH eye-specific $C_4$ isozyme. The isozymes were detected in the cytosol of eye tissues from Lepomis macrochirus and Micropterus salmoides and were more similar to the $A_4$ than the $B_4$ isozyme. LDH/CS in the eye tissue of L. macrochirus was increased in September, so the ratio of anaerobic metabolism was high. The electrophoretic patterns of mitochondrial LDH were similar to those of cytosolic LDH in the eye tissues of L. macrochirus and Micropterus salmoides. LDH eye-specific $C_4$ isozyme from eye tissue was purified by preparative native-PAGE. The activities of LDH eye-specific $C_4$ isozymes in L. macrochirus and M. salmoides were reduced at concentrations greater than 0.2 mM and 0.1 mM of pyruvate, respectively. These concentrations remained at 5.2% and 15.8% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The LDH activities of eye tissues were reduced at concentrations greater than 22 mM and 24 mM of lactate, respectively, in L. macrochirus and M. salmoides. The ${K_m}^{PYR}$ of eye-specific $C_4$ was 0.088 mM in L. macrochirus and it was 0.033 mM in M. salmoides. The activities of cytosolic and mitochondrial eye-specific $C_4$ isozymes were high in ${\alpha}$-ketobutyric acid. Furthermore, the activities of eye tissue and eye-specific $C_4$ isozyme had to be measured with 0.5 mM of pyruvate and a buffer solution of pH 7.5. As a conclusion, the eye-specific $C_4$ isozyme in M. salmoides has a high affinity for pyruvate and exhibits maximum activity at a lower concentration of pyruvate and at higher concentration of lactate than that in L. macrochirus. Therefore, it seems that the energy produced by the LDH eye-specific $C_4$ isozyme in M. salmoides was used at the first stage of predatory behavior.

Changes in Inorganic Element Concentrations in Leaves, Supplied and Drained Nutrient Solution according to Fruiting Node during Semi-forcing Hydroponic Cultivation of 'Bonus' Tomato ('Bonus' 토마토 반촉성 수경재배 시 착과절위에 따른 식물체, 공급액 및 배액의 무기성분 농도 변화)

  • Lee, Eun Mo;Park, Sang Kyu;Lee, Bong Chun;Lee, Hee Chul;Kim, Hak Hun;Yun, Yeo Uk;Park, Soo Bok;Chung, Sun Ok;Choi, Jong Myung
    • Journal of Bio-Environment Control
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    • v.28 no.1
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    • pp.38-45
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    • 2019
  • Recycling of drained nutrient solution in hydroponic cultivation of horticultural crops is important in the conservation of the water resources, reduction of production costs and prevention of environmental contamination. Objective of this research was to obtain the fundamental data for the development of a recirculation system of hydroponic solution in semi-forcing cultivation of 'Bonus' tomato. To achieve the objective, tomato plants were cultivated for 110 days and the contents of inorganic elements in plant, supplied and drained nutrient solution were analyzed when crop growth were in the flowering stage of 2nd to 8th fruiting nodes. The T-N content of the plants based on above-ground tissue were 4.1% at the flowering stage of 2nd fruiting nodes (just after transplanting), and gradually get lowered to 3.9% at the flowering stage of 8th fruiting nodes. The tissue P contents were also high in very early stage of growth and development and were maintained to similar contents in the flowering stage of 3rd to 7th fruiting nodes, but were lowed in 8th node stages. The tissue Ca, Mg and Na contents in early growth stages were lower than late growth stages and the contents showed tendencies to rise as plants grew. The concentration differences of supplied nutrient solution and drained solution in $NO_3-N$, P, K, Ca, and Mg were not significant until 5 weeks after transplanting, but the concentration of those elements in drained solution rose gradually and maintained higher than those in supplied solution. The concentrations of B, Fe, and Na in drained solution were slightly higher in the early stages of growth and development and were significantly higher in the mid to late stages of growth than those in supplied solution. The above results would be used as a fundamental data for the correction in the inorganic element concentrations of drained solution for semi-forcing hydroponic cultivation of tomato.

Soil Microbial Flora and Chemical Properties as Influenced by the Application of Pig Manure Compost (돈분퇴비의 시용이 배추재배지 토양의 미생물상 및 화학성에 미치는 영향)

  • Weon, Hang-Yeon;Kwon, Jang-Sik;Suh, Jang-Sun;Choi, Woo-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.1
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    • pp.76-83
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    • 1999
  • Studies were conducted during 2 months from May of 1997 to evaluate the effects of pig manure compost(PMC) on soil microbial flora. To do so, a field experiment of Chinese cabbage(Brassica campestris L.) was conducted in a randomized block design on a sandy loam soil and microbial floral characteristics in soils were analyzed. Treatments to control included the application of PMC at (A) $8Mg\;ha^{-1}$CM-8), (B) $29Mg\;ha^{-1}$(CM-2,9), and (C) $57Mg\;ha^{-1}$(CM-57), and of chemical fertilizer(D) at $320N-80P_2O_5-200K_2O\;kg\;ha^{-1}$(NPK). In each treatment, the rhizosphere and non-rhizosphere soils were tested for the analysis of microbial populations. The populations of bacteria, actinomycetes, and fungi increased in soils with the applications of PMC and chemical fertilizer, but that of Bacillus sp. decreased. However, the population of fluorescent Pseudomonas sp. was reduced in NPK plots only. With increasing application rates of PMC, the number of colony forming units(cfu) of bacteria (Pseudomonas sp. and actinomycetes) and fungi increased. in all PMC-treated plots, the population density peaked at early growth stage for bacteria(including Bacillus sp.), at late growth for fluorscent Pseudomonas sp., and at harvest for fungi and actinomycetes. The rhizosphere effect was greatest for fluorscent Pseudomonas sp. As the application rates of PMC increased, Total N, organic matter, available phosphate, and exchangeable -K, -Ca, and -Mg increased compared to control, but soil pH was lowered. In NPK plots, EC was 3.4-fold and exchangeable K was 5-fold higher than control.

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Characteristics of Manure and Estimation of Nutrient and Pollutant of Holstein Dairy Cattle (홀스타인 젖소 분뇨의 특성과 비료성분 및 오염물질 부하량 추정)

  • Choi, D.Y.;Choi, H.L.;Kwag, J.H.;Kim, J.H.;Choi, H.C.;Kwon, D.J.;Kang, H.S.;Yang, C.B.;Ahn, H.K.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.137-146
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    • 2007
  • This study was conducted to determine fertilizer nutrient and pollutant production of Holstein dairy cattle by estimating manure characteristics. The moisture content of feces was 83.9% and 95.1% for urine. The pH of feces and urine were in the ranges of 7.0~7.4 and 7.5~7.8, respectively. The average BOD5, COD, SS, T-N, T-P concentrations of the dairy feces were 18,294, 52,765, 102,889, 2,575, and 457mg/ℓ, respectively. Dairy urine showed lower levels of BOD5(5,455mg/ℓ), COD(8,089mg/ℓ), SS(593mg/ℓ), T-N(3,401mg/l), and T-P(13mg/ℓ) than feces. The total daily produced pollutant amounts of a dairy cow were 924.1g(Milking cow), 538.8g(Dry cow), 284.4g(Heifer) of BOD5, 2,336.5g (Milking cow), 1,651.8g(Dry cow), 734.1g(Heifer) of COD and 4,210.1g(Milking cow), 2,417.1g(Dry cow), 1,629.1g(Heifer) of SS and 194.8g(Milking cow), 96.4g(Dry cow), 58.3g(Heifer) of T-N and 24.0g(Milking cow), 10.2g(Dry cow), 6.1g(Heifer) of T-P. The calculated amount of pollutants produced by a 450kg dairy cow for one year were 181.3kg of BOD5, 492.5kg of COD, 899.9kg of SS, 36.0kg of T-N and 4.1kg of T-P. The total yearly estimated pollutant production from all head(497,261) of dairy cattle in Korea is 90,149 tons of BOD5, 244,890 tons of COD, 447,491 tons of SS, 17,898 tons of T-N and 2,008 tons of T-P. The fertilizer nutrient concentrations of dairy feces was 0.26% N, 0.1% P2O5 and 0.14% K2O. Urine was found to contain 0.34% N, 0.003% of P2O5 and 0.31% K2O. The total daily fertilizer nutrients produced by dairy cattle were 197.4g (Milking cow), 97.4g(Dry cow), and 57.9g(Heifer) of Nitrogen, 54.2g(Milking cow), 22.2g(Dry cow), and 14.2g(Heifer) of P2O5 and 110.8g(Milking cow), 80.4g (Dry cow), and 39.5g(Heifer) of K2O. The total yearly estimated fertilizer nutrient produced by a 450kg dairy animal is 36.2kg of N, 8.8kg of P2O5, 24.6kg of K2O. The estimated yearly fertilizer nutrient production from all dairy cattle in Korea is 18,000 tons of N, 4,397 tons of P2O5, 12,206 tons of K2O. Dairy manure contains useful trace minerals for crops, such as CaO and MgO, which are contained in similar levels to commercial compost being sold in the domestic market. Concentrations of harmful trace minerals, such as As, Cd, Hg, Pb, Cr, Cu, Ni, Zn, met the Korea compost standard regulations, with some of these minerals being in undetected amounts.

The Scope and the Meaning of 'Time of Arrival' in Carriage of Passengers by Air : Focused on the Germanwings GmbH v. Ronny Henning, Case C-452/13 (2014). (항공여객운송에서의 지연보상과 도착시각의 의미 - EU 사법재판소 2014. 9. 14. 판결(ECLI:EU:C:2014:2141)을 중심으로 -)

  • Sur, Ji-Min
    • The Korean Journal of Air & Space Law and Policy
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    • v.33 no.2
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    • pp.267-290
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    • 2018
  • This paper reviews and criticizes the EU Case of C-452/13, Germanwings GmbH v. Ronny Henning. Under this case, Ronny Henning later sued Lufthansa's budget carrier Germanwings after it refused to pay him 250 euros compensation for a delay he said totalled more than three hours. Germanwings, however, maintained his flight had arrived only two hours and 58 minutes behind schedule. In those circumstances, the following question to the European Court of Justice (ECJ) for a preliminary ruling: What time is relevant for the term time of arrival used in Articles 2, 5 and 7 of Regulation [No 261/2004]: (a) the time that the aircraft lands on the runway (touchdown); (b) the time that the aircraft reaches its parking position and the parking brakes are engaged or the chocks have been applied (in-block time); (c) the time that the aircraft door is opened; (d) a time defined by the parties in the context of party autonomy? ECJ says that the situation of passengers on a flight does not change substantially when their aircraft touches down on the runway at the destination airport, when that aircraft reaches its parking position and the parking brakes are engaged or when the chocks are applied, as the passengers continue to be subject, in the enclosed space in which they are sitting, to various constraints. Therefore, it is only when the passengers are permitted to leave the aircraft and the order is given to that effect to open the doors of the aircraft that the passengers may in principle resume their normal activities without being subject to those constraints. ECJ rules that it is apparent that Articles 2, 5 and 7 of Regulation No 261/2004 must be interpreted as meaning that the concept of 'arrival time', which is used to determine the length of the delay to which passengers on a flight have been subject, corresponds to the time at which at least one of the doors of the aircraft is opened, the assumption being that, at that moment, the passengers are permitted to leave the aircraft.

Analysis of Ingredients and DPPH, ABTS Activity for the Development of Cosmetic Raw Materials using 5 Kinds of Plants Native to Mt. Jiri (지리산 자생식물 5종의 화장품 원료개발을 위한 성분 및 DPPH, ABTS 활성분석)

  • Youn Ok, Jung;Bo Kyung, Kang;No Bok, Park
    • Journal of Practical Agriculture & Fisheries Research
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    • v.24 no.4
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    • pp.18-29
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    • 2022
  • Five species of plants (Clerodendrum trichotomum Thunb., Angelica dahurica (Fisch. ex Hoffm.) Benth. & Hook. f. ex Franch. & Sav., Caryopteris incana (Thunb. ex Houtt.) Miq., Lonicera japonica Thunb., and Parasenecio auriculatus var. matsumurana Nakai) native to the clean area of Mt. Jiri were selected. The collection period was from May to September 2021, and the five species plants were collected in their native habitats with flowers in full bloom. The collected plants were extracted with 70% EtOH, and 17 kinds of polyphenol components were analyzed. Next, flowers, leaves, stems, and roots were separated from plants, extracted with 70% EtOH for each part and experiments were conducted on DPPH, ABTS, total polyphenols, and total flavonoids. The results are as follows. 1. It was found that there were a total of 8 kinds of polyphenols contained in 5 species of plants that are native to Mt. Jiri. Among the polyphenol components, chlorogenic acid was contained in 4 species of plants, and caffeic acid was contained in 2 species of plants. 2. As a result, the DPPH radical scavenging activity was the best in the stem of P. auriculata and the C. trichotomum was good regardless of the specific part. It was found that the activity-scavenging activity was good in the flowers of A. dahurica and the leaves of L. japonica. 3. The highest ABTS radical scavenging activity was C. trichotomum Thunb., whose EC50 value was 38.73~66.28ppm. Next, the leaves and stems of L. japonica Thunb., A. dahurica and P. auriculata, and the leaves and stems of C. incana appeared in that order. 4. The highest total polyphenol content was 154.83mg GAE/g in the leaves of C. trichotomum, followed by about 130mg GAE/g in the flowers of C. trichotomum and P. auriculata. The lowest was 26.27mg GAE/g in the stems of A. dahurica.

The Study of Solid Waste Compost Development for Reclaiming Damage Soil in Forest (산림훼손토양 복원을 위한 부숙토 개발 연구)

  • Na, Seung-Ju;Chang, Ki-Woon;Yang, Hui-Young;Jeon, Han-Ki;Lee, Jong-Jin
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.2
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    • pp.107-120
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    • 2005
  • To study the development of solid waste compost to use sewage sludge and paper mill sludge for reclaiming damage soil in forest, the changes of temperature, moisture, chemical properties, heavy metals and harmful compound during the aerobic decomposition were investigated, and the compost decomposition of final products investigated the round paper chromatography method and G.I(Germination index) value. The results were summarized as follows. Temperature was changed a little during early 5days because of air temperature too low. That was rapidly increased to over $50^{\circ}C$ at 4days after first turning and then decreased gradually fallen to $40{\sim}50^{\circ}C$ at 15days after aerobic decomposition in A and C treatments. The second turning was conducted at 18 days after aerobic decomposition, and then the temperature was little changed. At the compare first with terminal product, The moisture content was decreased all treatments but the change was little in A and B treatments. pH was decreased to below 1 in all treatments. EC was increased to below 5dS/m. The content of total carbon, C/N ratio, $NH_4{^+}-N$ were decreased with 4~7%, below 8 and below 500mg/kg in all treatments, respectively. The content of total nitrogen, $NO_3{^-}-N$, CEC were increased with below 0.5%, below 173mg/kg and over $30cmol^+/kg$ in all treatments, respectively. The content of heavy metals and harmful compound were similar during aerobic decomposition and suited to standard of 가 grade in all treatments. The result of round paper chromatography method and G.I. value, The C treatment concluded well aerobic decomposition. Especially, the G.I. value in C treatment was 64.1 and 66.2 at cabbage and grass, respectively.

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Biological activity of shizukanols isolated from Chloranthus japonicus roots (홀아비꽃대(Chloranthus japonicus) 뿌리로부터 분리한 shizukanol들의 생물활성)

  • Park, Mi-Ran;Kim, Hae-Young;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Kim, Jin-Seog;Hong, Kyung-Sik;Park, No-Joong;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Pesticide Science
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    • v.8 no.4
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    • pp.338-346
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    • 2004
  • The methanol extract of Chloranthus japonicus roots effectively controlled the development of rice blast(Magnaporthe grisea), rice sheath blight(Corticium sasaki), tomato pay mold(Btrytis cinerea), tomato late blight(Phytophthora infestans), and wheat leaf rust(Puccinia recondita). From the methanol extract of C. japonicus roots, three antifungal substances were isolated. Their chemical structures were determined to be shizukanols B, C, and D mainly by mass and NMR spectral data. Among the three substances, shizukanol C showed the strongest inhibitory activity against mycelial growth of the plant pathogenic fungi tested; it completely inhibited mycelial growth of M. grisea. Colletotrichum gloeosporioides, and C. acutatum at concentrations of more than $12.5{\mu}g$/ and P. infestans at concentrations of more than $3.13{\mu}g/m\ell$. They also controlled effectively the development of rice blast and wheat leaf rust. On the other hand, they caused phytotoxic symptoms on barley leaves and inhibited the growth of duckweed (Lemna paucicostata) with $EC_{50}$ values of $30.0{\mu}g/m\ell$ for shizukanol B, $49.9{\mu}g/m\ell$ for shizukanol C, and $154{\mu}g/m\ell$ for shizukanol D. In addition, shizukanol C showed an insecticidal activity against brown planthopper (Nilaparavata lugens), peen peach aphid (Myzus persicae), diamond-back moth (Plutella xylostella), and tobacco cutworm (Spodoptera litura) of the 5 arthropod pests tested with mortality values of more than 60% at a concentration of $1,000{\mu}g/m\ell$.

Effect of Microorganism Mixture Application on the Microflora and the Chemical Properties of Soil and the Growth of Vegetables in Greenhouse (미생물혼합제제 처리가 토양의 미생물상과 화학적 특성 및 시설 채소 생육에 미치는 영향)

  • Ryu, Il-Hwan;Jeong, Su-Ji;Han, Seong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.31 no.4
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    • pp.368-374
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    • 2012
  • BACKGROUND: The urgency of feeding the world's growing population while combating soil pollution, salinization and desertification requires suitable biotechnology not only to improve crop productivity but also to improve soil health through interactions of soil nutrient and soil microorganism. Interest in the utilization of microbial fertilizer has increased. A principle of nature farming is to produce abundant and healthy crops without using chemical fertilizer and pesticides, and without interrupting the natural ecosystem. Beneficial microorganisms may provide supplemental nutrients in the soil, promote crop growth, and enhance plant resistance against pathogenic microorganisms. We mixed beneficial microorganisms such as Bacillus sp. Han-5 with anti-fungal activities, Trichoderma harziaum, Trichoderma longibrachiatum with organic material degrading activity, Actinomycetes bovis with antibiotic production and Pseudomonas sp. with nitrogen fixation. This study was carried out to investigate the mixtures on the soil microflora and soil chemical properties and the effect on the growth of lettuce and cucumber under greenhouse conditions. METHODS AND RESULTS: The microbial mixtures were used with each of organic fertilizer, swine manure and organic+swine manure and compared in regard to changes in soil chemical properties, soil microflora properties and crop growth. At 50 days after the treatment of microorganism mixtures, the pH improved from 5.8 to 6.3, and the EC, $NO_3$-Na and K decreased by 52.4%, 60.5% and 29.3%, respectively. The available $P_2O_5$ and $SiO_2$ increased by 25.9% and 21.2%, respectively. Otherwise, the population density of fluorescent Pseudomonas sp. was accelerated and the growth of vegetables increased. Moreover, the population density of E. coli and Fusarium sp., decreased remarkably. The ratio of bacteria to fungi (B/F) and the ratio of Actinomycetes bovis to fungi (A/F) increased 2.3 (from 272.2 to 624.4) and 1.7 times (from 38.3 to 64), respectively. Furthermore, the growth and yield of cucumber and lettuce significantly increased by the treatment of microorganism mixtures. CONCLUSION(S): These results suggest that the treatment of microorganism mixtures improved the chemical properties and the microflora of soil and the crop growth. Therefore, it is concluded that the microorganism mixtures could be good alternative soil amendments to restore soil nutrients and soil microflora.