• Title/Summary/Keyword: Agar concentration

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THE COMPARISON OF STREPTOCOCCUS MUTANS ISOLATED FROM OCCLUSAL SURFACES OF CARIES AND NON-CARIES TEETH (우식치아와 정상치아의 교합면에서 분리한 Streptococcus mutans의 비교)

  • Park, Ho-Won;Jung, Tae-Sung;Jung, Jin;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.1
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    • pp.129-141
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    • 2001
  • When oral microorganisms were sampled from occlusal surfaces of caries and non-caries teeth, $3.43\times10^5$ CFU and $3.47\times10^3$ CFU of bacteria were counted on MSB agar plates, respectively. All the 20 colonies isolated from a caries surface were Streptococcus mutans but, only two of 20 colonies were identified as Streptococcus mutans by API test. S. mutans SM1 from caries tooth and S. mutans SM2 from non-caries tooth showed the same results except for $\alpha-galactosidase$ activity on sugar fermentation tests and biochemical tests. For the bacterial replication, both SM1 and SM2 were actively multiplicated at pH 5.5. And the viability of SM1 was high at 20% of sucrose, while that of SM2 was high at 5% of sucrose in the media. SM1 actively replicated at 16mM of $CaCl_2$, 160mM of KCl, and 6.4mM of $MgCl_2$, and the replication of SM2 was increased at 16mM of $CaCl_2$, 40mM of KCl, 6.4mM of $MgCl_2$. At 1mM of sodium bicarbonate and sodium phosphate, both bacteria were actively multiplicated. SM1 and SM2 were actively replicated at 1mM and 10mM of Tris, respectively. For potassium phosphate buffer, SM1 grew well proportionally to the concentration up to 100mM, while the growth of SM2 were inhibited by the increase of concentration. The 4.6 kb of gtf gene was amplified with a pair of primer, gtfB-F961 and gtfC-R5574 by polymerase chain reaction from the chromosomal DNA of SM1 and SM2. When 4.6kb bands were eluted from gel and were treated with restriction enzyme, EcoR I produced the same RFLP like 0.8kb and 3.8kb of DNA fragments for S. mutans GS-5, SM1 and SM2. By Hind III, the PCR products weren't digested for S. mutans GS-5 and SM1, but 3 fragments such as 2.4kb, 1.8kb and 400bp were examined for SM2. These results indicated the difference between gtf genes of SM1 and SM2. BamH I treatment showed 4 fragments for SM1 and SM2, while the 3 fragments for S. mutans GS-5. The PCR products were not digested by Kpn I, Sma I, Xho I and Pst I.

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Elucidation of Mode of Action of Pantoea agglomerans 59-4 for Controlling Garlic Blue Mold (마늘 푸른곰팡이병 방제용 Pantoea agglomerans 59-4의 억제기작 해석)

  • Kim, Yong-Ki;Kwon, Mi-Kyung;Yeh, Wan-Hae;Hong, Sung-Jun;Jee, Hyung-Jin;Park, Jong-Ho;Han, Eun-Jung;Park, Kyung-Seok;Lee, Sang-Yeob;Lee, Seong-Don
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.163-169
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    • 2010
  • To screen for potential biocontrol agents against postharvest disease of garlics caused by Penicillium hirsutum, a total of 1292 isolates were isolated from the rhizoshere or rhizoplane of Allium species. Among them, S59-4 isolate was selected as a potential biocontrol agent by in vivo wounded garlic bulb assay. The isolate was identified as Pantoea agglomerans (Pa59-4) through Biolog system. Pa59-4 did not inhibit the mycelial growth of P. hirsutum in dual-culture with P. hirsutum on tryptic soy agar. In order to elucidate mode of action of Pa59-4 on biological control, nutrient competition between Pa59-4 and P. hirsutum was investigated by the simple method using tissue culture plates with cylinder inserts containing defusing membrane reported by Janisiewicz et al. (2000). The results showed that Pa59-4 effectively suppressed spore germination and mycelial growth of blue mold in the low concentration (0.5%) of garlic juice, but it did not suppress those of blue mold in the high concentration (5%) of garlic juice. This result suggests that the mechanism in biocontrol of garlic blue mold by Pa 59-4 may be involved in nutrient competition with P. hirsutum on garlic bulbs.

DETECTION OF METHICILLIN OR VANCOMYCIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM DENTAL HOSPITAL (치과병원 진료실 내에서 메티실린 또는 반코마이신 저항성 Staphylococcus aureus의 검출)

  • Min, Jung-Hee;Park, Soon-Nang;Hwang, Ho-Keel;Min, Jung-Beum;Kim, Hwa-Sook;Kook, Joong-ki
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.102-110
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    • 2007
  • The purpose of this study was to obtain the basic information for the improvement of dental environment by investigating the presence of methicillin- or vancomycin-resistant Staphylococcus aureus (MRSA or VRSA) isolated from dental health care workers (DHCWs) and environment of the Chosun University Dental Hospital (CUDH) and a private dental clinic (control group). Staphylococcus aureus (S. aureus) was isolated from anterior nares of 42 DHCWS and 38 sites, unit chairss, x-ray devices, computers, etc., at 10 departments of the CUDH and 20 DHCWs and 11 sites at the private dental clinic. S. aureus was isolated on mannitol salt agar plate and confirmed by PCR with S. aureus species-specific primer. Antimicrobial susceptibility test of clinical isolates of S. aureus against several antibiotics including methicillin (oxacillin) was performed by investigating minimum inhibitory concentration (MIC) using broth microdilution assay. In addition, PCR was performed to detect the methicillin- or vancomycin-resistant gene. The data showed that one strain of S. aureus was isolated from DHCWs of the CUDH and three strains of S. aureus was isolated from 3 samples of the private dental clinic, respectively. All of the isolates from the CUDH and the private dental clinic had resistance to penicillin G, amoxicillin and vancomycin and susceptibility to oxacillin and ciprofloxacin. The S. aureus strains were already obtained the resistance to penicillin G and amoxicillin. These results suggest that two dental clinics were under relatively safe environment.

Influence of $TiO_2$ Concentrations and Irradiation Lights on the Photocatalytic Reaction for Inhibiting Growth of Streptococcus mutans ($TiO_2$ 농도 및 여기광에 따른 광촉매 반응이 Streptococcus mutans의 생장에 미치는 영향)

  • Kang, Si-Mook;Lee, Hae-Na;Kim, Hee-Eun;Kim, Baek-Il
    • Journal of dental hygiene science
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    • v.14 no.3
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    • pp.319-324
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    • 2014
  • The aim of this study was to evaluate influences of titanium dioxide ($TiO_2$) concentrations and irradiation times on growth of Streptococcus mutans when irradiated by visible light (405 nm wavelength) and by ultraviolet light (254 nm wavelength). To find the optimal antibacterial concentration of $TiO_2$, 0.01, 0.1, 1.0, and 10.0 mg/ml $TiO_2$ suspension was prepared with sterilized distilled water. S. mutans cultured media was added to $TiO_2$ solution to set the final cell count to $10^4CFU/ml$. The photocatalytic reaction was induced by irradiating 254 nm and 405 nm lights for 10 minutes. To compare the bactericidal activities according to irradiation times, all photocatalytic reaction was carried out with 0.1 mg/ml $TiO_2$ for 0, 10, 20, 30, and 40 minutes with both lights. After the photocatalytic reaction, $100{\mu}m$ of the reaction mixture was immediately plated on brain heart infusion agar. These plates were placed at 5% $CO_2$, $37^{\circ}C$, for 24 hours and the bacterial colonies were counted. All experiments were performed in quintuplicate. One-way ANOVA was used to determine whether there were any significant differences between the $TiO_2$ concentrations or the irradiation times. The most effective concentration of $TiO_2$ for its photocatalytic bactericidal effect on S. mutans was 0.1 mg/ml when irradiated with 254 nm and 405 nm lights. The longer the irradiation time, the bigger the bactericidal effect for both wavelengths. Over 99% of bacteria in the inoculum were killed after irradiation with 254 nm for 20 minutes and with 405 nm for 40 minutes. In conclusion, a photocatalytic reaction of $TiO_2$ induced by visible light of 405 nm constitutes the bactericidal effect on S. mutans.

Herbicidal Activity of Wood Vinegar from Quercus mongolica Fisch (신갈나무 (Quercus mongolica Fisch) 목초액의 제초활성)

  • Kim, Song-Mun;Kim, Yong-Ho;Kim, Jin-Seog;Ahn, Mun-Sub;Heo, Su-Jeong;Hur, Jang-Hyun;Han, Dae-Sung
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.82-88
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    • 2000
  • The objective of this study was to determine if wood vinegar of Quercus mongolica Fisch has herbicidal activity. Growth of plants, such as barnyard grass (Echinochloa crus-gulli P. Beauv), quackgrass (Agropyron smithii RYDB), canola (Brassica napus L.), velvetleaf (Abutilon avicennae), indian jointvetch (Aeschynomene indica), and common sorghum (Sorghum bicolor) grown on agar batch treated with 0.01% wood vinegar were similar to that of plant without wood vinegar. The growth of such plants, however, reduced at $0.1{\sim}1%$ concentrations, and inhibited totally at >5% concentration. In greenhouse study, soil-applied wood vinegar did not inhibit tile growth of canola, barnyard grass, large crabgrass, and Abutilon avicennae even at the highest concentration, 80L $80L^{-1}\;10a^{-1}$, while foliar-applied wood vinegar did inhibit the growth of plants at higher than 40L $80L^{-1}\;10a^{-1}$. Growth of canola, barnyard grass, large crabgrass, and Abutilon avicennae treated with wood vinegar (80L $80L^{-1}\;10a^{-1}$) was reduced by 71, 46, 24, and 47%, respectively. In field experiment conducted at Chunchon and Taebeck, biomass of weeds treated with wood vinegar at less than 40L $80L^{-1}\;10a^{-1}$ were close to that of weeds treated without wood vinegar, while biomass of weeds at 80L $80L^{-1}\;10a^{-1}$ was reduced by 34-36%, compared to that of control, at both sites. However, the herbicidal activity of wood vinegar was much lower than that of glyphosate. Results in this study show that wood vinegar of Quercus mongolica Fisch has herbicidal activity, although the herbicidal activity was lower than that of glyphosate, a commercial herbicide.

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Effect of Gamijipaesan Extracts against Mastitis Induced by Staphylococcus aureus Infection in a Rat Model through Anti-inflammatory and Antibacterial Effects (가미지패산(加味芷貝散)의 포도상구균 감염 유방염에 대한 항균활성 및 항염 효과)

  • Kwon, Ji-Myung;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.1
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    • pp.1-24
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    • 2013
  • Objectives: The object of this study was to observe the protective effect of Gamijipaesan aqueous extracts(GJS), which has been traditionally used in Korean medicine in obstetrics & gynecological fields as anti-infectious and anti-inflammatory agents, against mastitis induced by Staphylococcus aureus infection in a rat model through antibacterial, antiinflammatory, immunomodulatory, and anti-oxidant effects. Methods: Antibacterial activities of GJS against S. aureus were detected using standard agar microdilution methods, with the effects on the bacterial invasion and intracellular killing of individual test materials in human mammary gland carcinoma cell(MCF-7) and murine macrophages(Raw 264.7) at MIC1/2, MIC and MIC2 concentration levels. In addition, the effects on the cell viability, nitric oxide(NO), tumor necrosis factor(TNF)-${\alpha}$ and interleukin (IL)-6 productions of LPS activated Raw 264.7 cells. The changes on the mammary tissue viable bacterial numbers, myeloperoxidae(MPO), inducible nitric oxide synthetase(iNOS), TNF-${\alpha}$ and IL-6 contents were observed in the S. aureus in vivo intramammary infectious rat model. The anti-bacterial and anti-inflammatory effects were compared with ciprofloxacin and piroxicam, respectively in the present study. Results: MIC of GJS and ciprofloxacin against S. aureus were detected as $0.860{\pm}0.428$ (0.391-1.563) mg/ml and $0.371{\pm}0.262$(0.098-0.782) ${\mu}g/ml$, respectively. In addition, GJS and ciprofloxacin were also showed marked dosage-dependent inhibition of the both bacterial invasion and intracellular killing assays using MCF-7 and Raw 264.7 cells at MIC1/2, MIC and $MIC{\times}2$ concentrations, respectively. $ED_{50}$ against LPS-induced cell viabilities and NO, TNF-${\alpha}$ and IL-6 releases of GJS were detected as 0.72, 0.04, 0.08 and 0.11 mg/ml, and as 19.04, 4.18, 5.37 and 4.27 ${\mu}g/ml$ in piroxicam, respectively. 250 and 500 mg/kg of GJS also inhibit the intramammary bacterial growth, MPO, iNOS, TNF-${\alpha}$ and IL-6 contents in S. aureus in vivo intramammary infected rats, respectively. GJS 500 mg/kg showed quite similar antibacterial and anti-infectious effects as compared with ciprofloxacin 40 mg/kg and also showed similar anti-inflammatory effects as piroxicam 10 mg/kg, in S. aureus in vivo intramammary infectious models. Conclusions: The results obtained in this study suggest that over 250 mg/kg of GJS showed favorable anti-infectious effects against S. aureus infection in a rat model through their antibacterial, anti-inflammatory, immunomodulatory and anti-oxidant effects and therefore expected that GJS can be used as alternative therapies, having both anti-inflammatory and anti-infectious activities. However, more detail mechanism studies should be conducted in future with the efficacy tests of individual herbal composition of GJS and the screening of the biological active compounds in individual herbs. In the present study, GJS 500 mg/kg showed quite similar anti-infectious effects were detected as compared with ciprofloxacin 40 mg/kg treated rats, and also GJS shows quite similar anti-inflammatory effects as compared with piroxicam 10 mg/kg in S. aureus in vivo intramammary infectious rats, but ciprofloxacin did not showed any anti-inflammatory effects, and piroxicam did not showed anti-infectious effects in this study.

Characteristics of Rahnella aquatilis Strain AY2000 for an Anti-Yeast Substance Production (항효모성 물질 생산을 위한 Rahnella aquatilis AY2000 균주의 생육특성)

  • Kang, Min-Jung;Lee, Bok-Kyu;Kim, Kwang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.215-220
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    • 2008
  • Rahnella aquatilis AY2000 has an unique characteristic which produces an anti-yeast substance (AYS). The AYS of the strain AY2000 was always secreted on agar plate, however, its activity in liquid culture was labile upon storage of the medium. In this paper, cultural conditions of the strain AY2000 for the AYS production were investigated in liquid culture, and minimal inhibitory concentration (MIC) against Saccharomyces cerevisiae was determined for the AYS activity. MIC of the AYS cultured in PYG broth at $^25{\circ}C$ for 24 hr was $23.5{\mu}g/mL$, however, that in MYCS (pH 5.5) broth at the same condition was $15.5{\mu}g/mL$. The activity of the AYS had increased rather in MYCS broth excluded $NH_4$-citrate than in the same broth contained $NH_4$-citrate, and MIC of the AYS produced in MYCS broth without $NH_4$-citrate was $15.5{\mu}g/mL$. When the strain AY2000 was maintained in MYCS broth without $NH_4$-citrate but added $100{\mu}M$ $FeCl_3$, the activity of the AYS had increased and its MIC was $7.8{\mu}g/mL$. MIC of the AYS was $7.8{\mu}g/mL$ after the strain AY2000 was cultured in MYCS broth containing $100{\mu}M$ $FeCl_3$ without $NH_4$-citrate, however, its MIC was $31.3{\mu}g/mL$ after 48-60 hr culture in the same broth.

Development of In Vitro Seedling Screening Method for Selection of Resistant Rice Against Bakanae Disease (벼 키다리병 저항성 검정을 위한 기내 유묘 검정법 개발)

  • Lee, Yong-Hwan;Lee, Myeong-Ji;Choi, Hyo-Won;Kim, Sung-Taek;Park, Jin-Woo;Myung, Inn-Shik;Park, Kyung-Seok;Lee, Se-Weon
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.288-294
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    • 2011
  • Rice bakanae disease, caused by the fungus Fusarium fujikuroi, is one of the most important rice diseases and distributed widely in Asia. Resistance screening system in rice field had been established. However, the evaluation results of the system vary according to the environmental conditions when the test is conducted. To develop precise and rapid evaluation method of disease resistance of rice to bakanae disease, in vitro screening system was attempted in this study. The six cultivars namely, 'Nampyeongbyeo', 'Junambyeo', 'Chucheongbyeo', 'Samcheonbyeo', 'Odaebyeo' and 'Hwasinbyeo' were tested. They were planted onto MS agar medium (10 ml) in test tube ($450\;mm{\times}{\phi}30\;mm$) and incubated at $25^{\circ}C$ and $28^{\circ}C$ in growth chamber under 12 hr light condition. Symptoms of over growth appeared a few days after seeding and then seedling were withered 2-3 weeks after over growth. The disease symptoms such as leaf dryness on top of rice were appeared in the 'Nampyeongbyeo' from 28 days at the concentration of $10^5$ spores/ml culturing at $28^{\circ}C$ and then withered completely 35 days after seeding. Whereas the other varieties withered entirely 19-23 days after seeding. Using the in vitro seedling screening method, 72 rice varieties were investigated to select resistant cultivar. Finally, two resistant cultivars ('Nampyeongbyeo' and 'Inwolbyeo') and seven moderately resistant cultivars ('Hwadongbyeo', 'Seokjeongbyeo', 'Samgwangbyeo', 'Sampyeongbyeo', 'Nonghobyeo', 'Heukjinjubyeo' and, 'Joanbyeo') were selected. If in vitro seedling screening method was used for evaluation of bakanae disease resistance, it would be completed within 35 days after sowing of rice seed.

Distribution and Favorable Conditions for Mycelial Growth of Cordyceps pruinosa in Korea (한국에 자생하는 붉은자루동충하초(Cordyceps pruinosa)의 분포와 균사생장에 적합한 조건)

  • Shin, Jae-Chul;Shrestha, Bhushan;Lee, Won-Ho;Park, Young-Jin;Kim, Soo-Young;Jeong, Gwang-Ryel;Kim, Ho-Kyung;Kim, Tae-Woong;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.79-88
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    • 2004
  • Cordyceps pruinosa grows upon dead pupae of Lepidoptera and produces one or $3{\sim}4$ club-shaped stromata per host. The stromata have distinct club-shaped head and long stalk. The length of stromata varies from $1{\sim}3\;cm$. Apical head consists of densely crowded semi-immersed perithecia, which are $360{\sim}400\;{\times}\;180{\sim}200\;{\mu}m$ in size. Asci are $150\;{\mu}m$ in length and $2.8{\sim}3\;{\mu}m$ in diameter. Ascospores, which are $124{\sim}141\;{\mu}m$ in length, have thin thread-like structures in the middle with part-spores attached on both sides. Each ascospore does not separate into part-spores after dispersal, but each part-spore germinates and together develops a colony. The imperfect form produces phialides of $15{\sim}24\;{\times}\;2{\sim}3\;{\mu}m$ size, with spherical or spindle shaped conidia of $4{\sim}6\;{\times}\;1.8{\sim}2.4\;{\mu}m$ size, The anamorph was identified as Mariannaea elegans Samson. YMA and SDAY agar media with pH 7 was produced abundant mycelial growth with high density. Best mycelial growth was observed when dextrin was used as a carbon source. Lactose, saccharose and sucrose also produced high mycelial growth. Peptone, yeast extract and tryptone produced abundant mycelial growth, when used as nitrogen sources. Highest mycelial growth and density was observed when C/N ratio was 1 : 1 at the concentration of 12.5 g/l each. $KH_2PO_4$ was the best mineral source for mycelial growth. Highest mycelial dry wt. was produced in YM and SDAY broths. Optimum inoculum for 100 ml of liquid broth was 6 mycelial discs. Similarly, optimum liquid culture period was 7 days.

Control of Phythophthora capsici and residual characteristics by drenching of pesticides on tomato in hydroponic culture system (약제 관주처리에 의한 양액재배 토마토의 역병 방제 및 농약잔류 특성)

  • Ihm, Yang-Bin;Lee, Jung-Sup;Kyung, Kee-Sung;Kim, Chan-Sub;Oh, Kyeong-Seok;Jin, Yong-Duk;Lee, Byung-Moo
    • The Korean Journal of Pesticide Science
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    • v.6 no.4
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    • pp.287-292
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    • 2002
  • To establish effective and safe control method against Phytophthora root rot caused by Phytophthora capsici on tomato in hydroponic culture, three pesticides, oxadixyl copper hydroxide 8% WP, metalaxyl copper oxychloride 15% WP, and dimethomorph. dithianon 38% WP at 4 concentration levels were tested on potato dextrose agar medium inoculated with Phytophthora capsici. All pesticides inhibited mycelial growth, but two pesticides of them, metalaxyl copper oxychloride WP and dimethomorph. dithianon WP, were selected as effective pesticides for the efficacy test in a hydroponic culture. Forty days after transplanting of tomato seedlings, 4 ml of sporangia of P. capsici (about 25 sporangi/ml) per plot was inoculated around tomato plant root, and then 5 days after inoculation, the pesticides diluted at 5,000 times were drenched 1, 2 or 3 times per plot on the culture cube at 15 days interval. Fifteen days after drenching, tomato fruits and hydroponic culture solution were sampled for the analysis of pesticide residues. Dimethomorph was detected 0.001 and 0.003 mg/kg in tomato of the plots sprayed 2 and 3 times with dimethomorph dithianon WP of which detection levels were far below compared with 1.0 mg/kg of the Korean MRL of dimethomorph on tomato. Incidences of Phytophthora root rot were $30.5{\sim}50%$ in the plots drenched at 1 or 2 times with metalaxyl.copper oxychloride WP, and $16.7{\sim}25%$ in the plots treated with dimethomorph dithianon WP. However, there was no incidence of Phytophthora root rot in the plots treated at 3 times with both of pesticides, showing no phytotoxic effect. Based on the results, the drenching of these pesticides on the culture cube could be recommended as a very safe and effective control method for Phytophthora root rot in tomato.