• 제목/요약/키워드: Inoculation method

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Polymerase chain reaction for the detection of Toxoplasma gondii in the blood of cats (중합효소연쇄반응(PCR)을 이용한 고양이 혈액내에서의 Toxoplasma gondii 검출에 관한 연구)

  • Suh, Myung-deuk;Joo, Bo-hyun
    • Korean Journal of Veterinary Research
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    • v.39 no.6
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    • pp.1151-1160
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    • 1999
  • This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

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Mutation and Selection of Agrobacterium tumefaciens Arginine Auxotroph by UV Irradiation (자외선 조사(照射)에 의한 Agrobacterium tumefaciens Arginine 요구주(要求株)의 유도와 선발)

  • Lee, Yearn;Park, Ro-Dong;Kim, Kwang-Sik
    • Applied Biological Chemistry
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    • v.28 no.2
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    • pp.92-97
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    • 1985
  • UV irradiation method was applied to Agrobacterium tumefaciens A 136 to obtain arginine auxotrophic mutant which is applicable as a host of Ti-plasmid. When the bacterial growth was measured at 600 nm, it showed the exponential phase between 7 and 16 hours after 2% inoculation (v/v) in TY medium and the generation time of 4.8 hours. Survival rate of $1{\sim}0.1%$ was reserved when irradiated at the intensity of $800\;{\mu}w/cm^2$ for $30{\sim}50sec$. Fifteen mutants were selected among 5,000 colonies after UV irradiation. Two of them were identified as arginine auxotrophs, three of them as asparagine auxotrophs, ana the other not as arginine, asparagine, glycine nor cysteine auxotrophs.

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Optical Monitoring of Tumors in BALB/c Nude Mice Using Optical Coherence Tomography

  • Song, Hyun-Woo;Lee, Sang-Won;Jung, Myung-Hwan;Kim, Kye Ryung;Yang, Seungkyoung;Park, Jeong Won;Jeong, Min-Sook;Jung, Moon Youn;Kim, Seunghwan
    • Journal of the Optical Society of Korea
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    • v.17 no.1
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    • pp.91-96
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    • 2013
  • We report a method for optical monitoring of tumors in an animal model using optical coherence tomography (OCT). In a spectral domain OCT system, a superluminescent diode light source with a full width of 66 nm at half maximum and peak wavelength of 950 nm was used to take images having an axial resolution of 6.8 ${\mu}m$. Cancer cells of PC-3 were cultured and inoculated into the hypodermis of auricle tissues in BALB/c nude mice. We observed tumor formation and growth at the injection region of cancer cells in vivo and obtained the images of tumor mass center and sparse circumferences. On the $5^{th}$ day from an inoculation of cancer cells, histological images of the tumor region using cross-sectional slicing and dye staining of specimens were taken in order to confirm the correlation with the high resolution OCT images. The OCT image of tumor mass compared with normal tissues was analyzed using its A-scan data so as to obtain a tissue attenuation rate which increases according to tumor growth.

Influence of Ozonated Water and Washing Method Using Ozonated Water for Controlling Food-borne Disease Bacteria (오존수와 오존수를 이용한 세척방법이 식중독 세균 제어에 미치는 영향)

  • Park, Jeongmi;Shin, Han-Seung
    • Food Engineering Progress
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    • v.14 no.4
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    • pp.316-321
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    • 2010
  • The efficacy of the ozonated water (0.1, 0.2, 0.4, 0.6, and 1.0 ppm) in reducing the risk of food-borne disease was investigated in this study. After inoculation of Listeria monocytogenes (ATCC 19112), Salmonella enterica subsp. enterica biovar Typhimurium (ATCC 12598), Escherichia coli O157:H7 (ATCC 43890) to lettuce, spinach, and beef, inhibition effect with different washing concentrations, time, and methods with ozonated and tap water were evaluated. As a result, there were 2.16 to 3.85 log CFU/mL reduction in different foods and 7 log CFU/mL reduction on cutting boards after watering with ozonated water. Higher than 0.2 ppm of ozonated water treatment reduced the growth of food-borne disease bacteria with increasing washing time and ozone concentration. These results suggested that the ozonated water treatment effectively improved the microbiological quality and food safety.

A Study on the Evaluation of Antimicrobial Effect of Orostachys Japonicus A. Berger Ethyl Acetate Fraction (와송 에틸아세테이트 분획물의 항균효능 평가에 관한 연구)

  • Im, Eun Kyung;Yang, Jae Chan
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.2
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    • pp.488-496
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    • 2021
  • In this study, a study was conducted to utilize Orostachys japonica A. Berger EtOAc fraction extract as an antibacterial activity and cosmetic ingredient. As a result of measuring the antimicrobial activity of Orostachys japonica A. Berger EtOAc, the growth of S. aureus, S. epidermidis, and P. aeruginosa was inhibited. Among them, S. aureus was an extract of 18.35 ± 1.5 mm Orostachys japonica A. Berger EtOAc fraction at a concentration of 0.5 g / mL, showing superior antibacterial activity than methyl paraben (16.83 ± 1.0 mm), and was shown as a positive control. As a result of evaluating the MIC of the Orostachys japonica A. Berger EtOAc fraction extract through MIC measurement, the remaining strains excluding Candida. A showed a MIC of 17.5 mg/mL. As a result of evaluating the cosmetic preservation effect through the challenge test applied to the cosmetic emulsion formulation, the growth inhibitory effect of S. aureus in the emulsion containing 0.3% Orostachys japonica A. Berger EtOAc fraction extract 7 days after microbial inoculation was 100%.

Immune gene expression and protection effect against VHSV by injection of interferon regulatory factor 10 in zebrafish (Danio rerio) (제브라피쉬 interferon regulatory factor 10의 주사에 따른 면역 유전자 발현과 VHSV에 대한 방어 효과)

  • Kim, Hye Ji;Kim, Jin Young;Park, Jong Bin;Lee, Ji Hyun;Park, Jeong Su;Kim, Hyoung Jun;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.23-29
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    • 2021
  • Interferon regulatory factors (IRFs) are a family of transcription factors essential to the control of antiviral immune response, cell growth, differentiation and apoptosis. IRF10 of zebrafish (Danio rerio) was negative regulation of the interferonΦ1 and 3 response in vitro. In this study, we analyze the induction of in vivo immune response activation from the IRF10 gene of zebrafish and the protective effect against VHSV. As the results, the group inoculated with IRF10 expression vectors, there was no expression of IFNΦ1, suggestion that IRF10 may function as a negative regulator of IRF3, which binds to the IFNΦ1 promoter. And other types of interferon genes (IFNΦ2-4) are thought to have been activated, inducing to the expression of pro-inflammatory cytokine and Mx genes. As the results of challenge test performed at 14 days after inoculation of the expression vectors, the maximum survival rate [50% (1㎍ DNA) and 42.5% (10㎍ DNA)] for IRF10 group were recorded. Meanwhile, the survival rates of pcDNA3.1 and PBS as the control groups were 10% and 15%, respectively. This study suggests that the possibility that activation of IRF10 molecule could be exploited as a VHS control method.

Efficacy of live NNV immersion vaccination in three grouper species (Seven band grouper Hyporthodus septemfasciatus, Red spotted grouper Epinephelus akaara, Long tooth grouper Epinephelus bruneus) (바리과 어류 3종(능성어 Hyporthodus septemfasciatus, 붉바리 Epinephelus akaara, 자바리 Epinephelus bruneus)에 대한 신경괴사증바이러스(Nervous Necrosis Virus, NNV)의 생백신 처리 효과)

  • Kim, Si-Woo;Park, Jung-Jun;Hong, Chang-Gi;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.99-104
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    • 2021
  • In this study, we investigated safety and efficacy of a low temperature immunization protocol with NNV in red spotted grouper, Epinephelus akaara and long tooth grouper, Epinephelus bruneus and seven band grouper, Hyporthodus septemfasciatus. Further, growth rate between immunized and naïve fish was evaluated during the experiment to check side effect of immunization. Three grouper species were immunized by immersion method with live NNV at 105.0 TCID50/mL at 16.5℃ for 30 min and reared for 120 days at natural sea water temperature. To evaluate growth rate, total length and wet weight was measured 7 times after immunization. Immunized three grouper species were challenged by intramuscular inoculation with NNV at 104.2 TCID50/100 µL/fish. Immunization at low temperature with live NNV did not show any clinical symptoms of infection, mortality and inhibition of growth. After challenge, cumulative mortality of naïve seven band grouper, red spotted grouper, long tooth grouper were 45, 10, 20 %, respectively. However no mortality was observed at immunized groupers. Thus, it was demonstrated that immunization at low temperature with live NNV are able to protect three different species of groupers without inhibition of growth.

Efficacy of Disinfectants against Health-Associated Multi-drug Resistant Clinical Isolates

  • An, Jeong-Lib;Kim, Sang-Ha;Yu, Young-Bin;Kim, Sunghyun;Lee, Moo-Sik;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.27 no.4
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    • pp.264-269
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    • 2021
  • The purpose of this study was to evaluate the disinfecting efficacy of peracetic acid (PAA), sodium hypochlorite (NaOCl) and phenol, which are representative disinfectants in medical environments using four types of multi-drug resistance (MDR) clinical isolates with healthcare-associated infections (HAI). 26 antibiotic susceptibility tests were conducted for the four types of MDR clinical isolates in the same way as for clinical specimens. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the disinfectants were determined by using in vitro liquid medium dilution method and inoculation of the plate medium. Both the MIC and MBC of phenol against MRSA and VRE were 3.1%, while those against KPC and MRPA were 6.2%. The MIC and MBC of peracetic acid (PAA) against MRSA, VRE, KPC, and MRPA were 0.18%. The MIC and MBC of sodium hypochlorite (NaOCl) against MRSA were 0.39% and 0.78%, respectively. Both values of MIC and MBC were 0.78% for VRE. In addition, KPC and MRPA showed 0.39% for MIC and 0.78% for MBC. For all MDR strains used in this study, sodium hypochlorite and peracetic acid showed significant sterilizing efficiency, while no clear correlation was identified between antibiotic resistance clinical isolated and ability of disinfection.

Characterization and Pathogenicity of Lasiodiplodia theobromae Causing Black Root Rot and Identification of Novel Sources of Resistance in Mulberry Collections

  • Gnanesh, Belaghihalli N.;Arunakumar, Gondi S.;Tejaswi, Avuthu;Supriya, M.;Manojkumar, Haniyambadi B.;Devi, Suvala Shalini
    • The Plant Pathology Journal
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    • v.38 no.4
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    • pp.272-286
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    • 2022
  • Black root rot (BRR) caused by Lasiodiplodia theobromae is an alarming disease of mulberry that causes tremendous economic losses to sericulture farmers in India and China. Successful control of this disease can be attained by screening germplasm and identifying resistant sources. Seventy four diseased root samples were collected from farmer's fields belonging to four major mulberry growing states of South India. Based on morpho-cultural and scanning electron microscopy studies, 57 fungal isolates were characterized and identified as L. theobromae. Phylogenetic analysis of concatenated internal transcribed spacer and β-tubulin sequences revealed variation of the representative 20 isolates of L. theobromae. Following the root dip method of inoculation, pathogenicity studies on susceptible mulberry genotypes (Victory-1 and Thailand male) recognized the virulent isolate MRR-142. Accordingly, MRR-142 isolate was used to evaluate resistance on a set of 45 diverse mulberry accessions. In the repeated experiments, the mulberry accession ME-0168 which is an Indonesian origin belonging to Morus latifolia was found to be highly resistant consistently against BRR. Eight accessions (G2, ME-0006, ME-0011, ME-0093, MI-0006, MI-0291, MI-0489, and MI-0501) were found to be resistant. These promising resistant resources may be exploited in mulberry breeding for developing BRR resistant varieties and to develop mapping populations which successively helps in the identification of molecular markers associated with BRR.

Survival Behavior of Escherichia coli O157:H7 during over 60-Day Aging of Camembert Cheeses Manufactured from Unpasteurized Raw Milk

  • Chon, Jung-Whan;Seo, Kun-Ho;Lim, Jong-Soo;Youn, Hye-Young;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.40 no.4
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    • pp.151-162
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    • 2022
  • Although many developed countries (USA, Canada, and several EU countries) allow raw milk cheese to be aged more than 60 days, these countries have strict standards for the aging conditions, such as temperature, of raw milk cheese. Spiking experiments were conducted with Camembert cheese made from raw milk, to assess the microbiological safety of raw milk cheese aged for more than 60 days. We spiked Escherichia coli O157:H7 into raw milk with different inoculation levels (high, medium, and low). Camembert cheese was prepared from the inoculated raw milk, then aged in an incubator for up to 9 weeks (63 days). There were no significant differences in pH and water activity (aW) between uninoculated cheese and cheese samples inoculated with E. coli O157:H7 (p<0.05). The pH and aWof the Camembert cheese decreased throughout the storage period. In conclusion, E. coli O157:H7 did not affect the pH and aW of the cheese samples. Cell counts were conducted every week using the agar-plating method. Inoculated cells were completely eliminated, especially in Camembert cheese, after 60 days, and the reduction rate of cells was much faster in Camembert cheese.