• Title/Summary/Keyword: strains inoculation

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Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran

  • Danehchin, Leila;Razmi, Gholamreza;Naghibi, Abolghasem
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.15-20
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    • 2016
  • Toxoplasmosis is an important zoonotic disease that can cause abortion in humans and animals. The aim of this study was isolation and subsequent genotyping of Toxoplasma gondii isolates in ovine aborted fetuses. During 2012-2013, 39 ovine aborted fetuses were collected from sheep flocks in Khorasan Razavi Province, Iran. The brain samples were screened for detection of the parasite DNA by nested PCR. The positive brain samples were bioassayed in Webster Swiss mice. The serum samples of mice were examined for T. gondii antibodies by IFAT at 6 weeks post inoculation, and T. gondii cysts were searched in brain tissue samples of seropositive mice. The positive samples were genotyped by using a PCR-RLFP method. Subsequently, GRA6 sequences of isolates were analyzed using a phylogenetic method. The results revealed that T. gondii DNA was detected in 54% (20/37, 95% CI 38.4-69.0%) brain samples of ovine aborted fetuses. In bioassay of mice, only 2 samples were virulent and the mice were killed at 30 days post inoculation, while the others were non-virulent to mice. The size of cysts ranged $7-22{\mu}m$. Complete genotyping data for GRA6 locus were observed in 5 of the 20 samples. PCR-RLFP results and phylogenetic analysis revealed that all of the isolated samples were closely related to type I. For the first time, we could genotype and report T. gondii isolates from ovine aborted fetuses in Khorasan Razavi Province, Iran. The results indicate that the T. gondii isolates are genetically related to type I, although most of them were non-virulent for mice.

Microbilolgical Studies on the Takju (Makguly) Brewing : The Korean Local Wine (한국 고유주의 일종인 막걸리에 대한 미생물학적 연구)

  • Koh, Choon-Myung;Choi, Tae-Joo;Lew, Joon
    • Korean Journal of Microbiology
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    • v.11 no.4
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    • pp.167-174
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    • 1973
  • This study included two parts of investigation, the microfloral changes during the brewing process with the changes of pH, total acidity, temperature and alcoholic contents, as well as determination of survival times of major enteric pathogens in Takju. 1. Maximum number of Saccharomyces cerevisiae was $4.3{\times}10^7$ per milliliter on the 5th day of fermentation and gradually decreased. Saccharomyces cerevisiae was one of the predominant strains of the fermentation process. The number of Saccharomyces cerevisiae was $4.3{\times}10^6$ per milliliter at the completion of the brewing and human consumption. In a few days after the completion of the brewing. Bacillus subtilis and some species of Staphylococcus spp. began to grow and those organisms were responsible for the spoilage. 2. Maximum pH, during the brewing, was 5.8 on the first day of fermentation and rapidly decreased until 6th day of fermentation at pH 4.3. 3. Maximum alcholic content was 14.5 degree on the 4th day of fermentation, 10.3 degree on the 5th day and this degree was continued during the experimentation. 4. Maximum temperature, during Takju brewing was 34.deg.C on the 3rd day of fermentation and rapidly decreased up to 23.deg.C on the 6th day and this temperature was continued until the brewing process was finished. 5. Maximum total acidity was 0.57 percent on the 4th day of fermentation and gradually decreased by brewing process was completed. 6. Survival time of major enteric pathogenic bacteria in Takju was as follows : Shigella dysenteriae and Escherichia coli were isolated in two hours and 14 hours respectively, but Salmonella typhi, Vibrio parahemolyticus were not isolated even in an hour after the inoculation of those organisms in undiluted Takju. In diluted Takju, Salmonella typhi, Vibrio parahemolyticus were not isolated even in an hour after the inoculation of those organisms in undiluted Takju. In diluted Takju, Salmonella typhi, Shigella dysenteriae, and Escherichia coli were survived for 50-60 hours, but Vibrio cholerae and Vibrio parahemolyticus were not isolated even if treated within one hour.

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Suppressing Erwinia carotovora Pathogenicity by Projecting N-Acyl Homoserine Lactonase onto the Surface of Pseudomonas putida Cells

  • Li, Qianqian;Ni, Hong;Meng, Shan;He, Yan;Yu, Ziniu;Li, Lin
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1330-1335
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    • 2011
  • N-Acyl homoserine lactones (AHLs) serve as the vital quorum-sensing signals that regulate the virulence of the pathogenic bacterium Erwinia carotovora. In the present study, an approach to efficiently restrain the pathogenicity of E. carotovora-induced soft rot disease is described. Bacillus thuringiensis-derived N-acyl homoserine lactonase (AiiA) was projected onto the surface of Pseudomonas putida cells, and inoculation with both strains was challenged. The previously identified N-terminal moiety of the ice nucleation protein, InaQ-N, was applied as the anchoring motif. A surface display cassette with inaQ-N/aiiA was constructed and expressed under the control of a constitutive promoter in P. putida AB92019. Surface localization of the fusion protein was confirmed by Western blot analysis, flow cytometry, and immunofluorescence microscopy. The antagonistic activity of P. putida MB116 expressing InaQ-N/AiiA toward E. carotovora ATCC25270 was evaluated by challenge inoculation in potato slices at different ratios. The results revealed a remarkable suppressing effect on E. carotovora infection. The active component was further analyzed using different cell fractions, and the cell surface-projected fusion protein was found to correspond to the suppressing effect.

Isolation and cultivation of swine encephalomyocarditis virus (돼지 뇌심근염 바이러스의 분리 배양)

  • Ha, Yong-kong;Yoon, Seok-min;Jung, Byung-tack;Park, Nam-yong;Lee, Bong-ju;Chung, Chi-young;Kee, Hye-young;Bae, Seong-yeol
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.479-484
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    • 1991
  • Encephalomyocarditis(EMC) virus was isolated from the mummified and stillborn pigs at a swine farm in Chonnam Province, experienced with EMC infection over the period Oct.~Dec. of 1989. In addition some cultural, serological properties of the isolates and experimental infections in the piglets were studied. The results obtained were as follows; 1. Two EMC virus strains with HA titers and CPE similar to EMC-ATCC were established in a baby hamster kidney (BHK)-21 cell line by inoculating homogenates of brain and heart of the 19 mummified or stillborn pigs and designated $K_3$ and $K_{11}$. 2. At the second BHK-21 cell line passage of the initial isolates CPE appeared after incubation for 16~18 hours, while at the fourth and fifth passage the highest titer of HA was recorded, titer of HA using rat and guinea pig erythrocytes. 3. One pig inoculated with the isolate $K_3$ showed dyspnea as clinical signs and died at the 10 days after inoculation at necropsy white necrotic foci were observed from the dead animal heart. 4. Although all the rest surviving pigs showed increases in antibody titer and body temperature of $40^{\circ}C$ above for the initial 2~4 days followed by the return to normal, there were no gross lesions when the animals were sacrificed at the 2 weeks after inoculation.

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Resistance Induction and Enhanced Tuber Production by Pre-inoculation with Bacterial Strains in Potato Plants against Phytophthora infestans

  • Kim, Hyo-Jeong;Jeun, Yong-Chull
    • Mycobiology
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    • v.34 no.2
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    • pp.67-72
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    • 2006
  • Efficacy of resistance induction by the bacterial isolates Pseudomonas putida (TRL2-3), Micrococcus luteus (TRK2-2) and Flexibacteraceae bacterium (MRL412), which were isolated from the rhizosphere of plants growing in Jeju Mountain, were tested in a greenhouse. The disease severity caused by Phytophthora infestans was effectively reduced in the potato plants pre-inoculated with bacterial isolates compared with those of the untreated control plants growing in a greenhouse. In order to estimate the level of protection by the bacterial isolates, Mancozeb WP (Diesen $M^{(R)}$, Kyong nong) and DL-3-amino butyric acid (BABA) were pre-treated, whereas Dimethomorph WP ($Forum^{(R)}$, Kyong nong) and phosphonic acid ($H_{3}PO_{3}$) were post-treated the challenge inoculation with the pathogen. Disease severities of chemical pre-treated as well as post-treated plants were reduced compare to those of the untreated. The disease reduction in the plants pre-treated with Mancozeb WP was the highest, whereas that of post-treated with Dimethomorph WP was the lowest. The yields of plants pre-inoculated with three bacterial isolates were greatly increased than those of control plants. These results suggest that biological control by bacterial isolates might be an alternative strategy against late blight disease in potato plants growing in greenhouse.

Optimal Condition for Deacidification Fermentation of Wild Grape Wine by Mixed Culture (혼합배양에 의한 산머루주의 감산발효 최적조건)

  • Kim, Seong-Ho
    • Applied Biological Chemistry
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    • v.51 no.1
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    • pp.17-23
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    • 2008
  • In order to prevent wine quality deterioration caused by strong sour taste from raw and other materials during fermentation of wild grape wine, the various mixed cultures conditions of the deacidification fermentation and the alcohol fermentation process by inoculation of mixed strains were investigated. As a result of mixed cultures process after the inoculation of Schizosaccharomyces pombe and Schizosaccharomyces japonicus with each deacidification fermentation strain in a culture of Saccharomyces sp. SMR-3 which was used in the alcohol fermentation strain of wild grape, cultures for 12 days at $22^{\circ}C$ with Saccharomyces sp. SMR-3 and Schizosaccharomyces pombe resulted in the maximum alcohol content at $15.8{\pm}0.2%$ and the minimum with the acidity of $0.44{\pm}0.02%$, the total organic acid of $648.96{\pm}7.14$ mg% and malic acid of $99.30{\pm}1.24$ mg%. Mixed cultures with Saccharomyces sp. SMR-3 and Schizosaccharomyces pombe under the optimal condition for the deacidification fermentation of wild grape wine showed 2% higher alcohol content, 51.65% lower acidity, 48.02% lower total organic acid, and 81.12% lower malic acid than a single culture of Saccharomyces sp. SMR-3.

Biofilm Formation Characteristics of Major Foodborne Pathogens on Polyethylene and Stainless Steel Surfaces

  • Kim, Hyeong-Eun;Kim, Yong-Suk
    • Journal of Food Hygiene and Safety
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    • v.35 no.2
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    • pp.195-204
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    • 2020
  • This research was investigated the effects of temperature and time against the formation of biofilms by foodborne pathogens on surfaces of polyethylene and stainless steel. After preliminary experiments with 32 strains from 6 species of foodborne pathogens (Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Salmonella Typhimurium), one strain from each species with the highest biofilm formation efficiency was selected. All foodborne pathogens showed a tendency toward an increased ability for biofilm formation with increasing temperature, but there was no consistency between the two materials and between foodborne pathogens. At all tested temperatures, the biofilm formation ability of E. coli and P. aeruginosa on the polyethylene surface was higher than that on the stainless steel surface with significant differences. The foodborne pathogens all formed biofilms immediately upon inoculation, and biofilm formation by E. coli, P. aeruginosa, and S. Typhimurium increased on both the polyethylene and stainless steel surfaces at 1 h after inoculation compared to at 0 h. At 7 days after biofilm formation, the other strains except S. aureus showed no difference in survival rates on polyethylene and stainless steel. The ability of these 6 foodborne pathogens to form biofilms showed different trends depending on the type of bacteria and the instrument material, i.e., polyethylene and stainless steel.

Plant Growth-promoting Ability by the Newly Isolated Bacterium Bacillus aerius MH1RS1 from Indigenous Plant in Sand Dune (해안 사구에서 서식하는 토착식물로부터 분리된 근권미생물 Bacillus aerius MH1RS1의 식물성장 촉진 능력 연구)

  • Lee, Eun Young;Hong, Sun Hwa
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.10
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    • pp.687-693
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    • 2013
  • Coastal sand dunes have been seriously damaged caused by the development thoughtless for the environment and coastal erosion and destruction due to artificial structures like coast roads and breakwater. Hereupon, in this study we made a library of rhizobacteria that have the plant growth-promoting ability for plant rhizosphere of indigenous plants inhabiting in a coastal sand dune as well as the strong tolerance to salt, and evaluated the plant growth-promoting ability of these strains. Furthermore, we evaluated the effect of rhizobacteria on the growth rate of saline tolerant plants in sandy soil; selected out the most useful micro-organism for the restoration of a damaged sand dune. The effect of inoculation of strains selected from the first experiment on the growth of Peucedanum japonicum and Arundo donaxes planted in a coastal sand dune was evaluated. As a result, Bacillus aerius MH1RS1 had plant growth promoting activities: indole acetic acid (IAA) production, siderophores and 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) activity, and also had a salinity tolerance. Also, in case of Peucedanum japonicum, the length of stems and weights of roots were enhanced by the inoculation of B. aerius MH1RS1. Fresh weights of stems and weights of roots in experimental group were, in particular, increased by 25% comparing with the control group. For an Arundo donax in experimental group, plant length increased by 18%, and weight of roots by 20% which is significant.

Processing of Radish Juice by Mixed Culture with lactic Acid Bacteria (유산균의 혼합배양에 의한 무주스의 가공)

  • 김정희;김종일
    • Food Science and Preservation
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    • v.6 no.4
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    • pp.448-455
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    • 1999
  • The starters, 0.3% of Leuconostoc mesenteroides, Lactobacillus brevis and Lactobacillus fermentum, were added in radish juice to process the radish juice by single and mixed cultures. The radish Juice was fermented for 7 days at 25$^{\circ}C$, 20 days at 15$^{\circ}C$, and 36 days at 5$^{\circ}C$. When fermented at 25$^{\circ}C$, the number of lactic acid bacteria in the juice made with mixed culture was higher than that of the single culture. But, juice fermented at low temperatures (15∼5$^{\circ}C$), the addition of starters was not effective, although there were some differences by inoculation strains. Although there was a little differences by inoculation strains, the content of nonvolatile organic acid and L-ascorbic acid were found more in the juice inoculated with lactic acid bacteria than the juice not inoculated. When the single and mixed cultures at the optimal maturity were tested, the significant difference was found at 5% level except the yeasty and moldy smell and the unripe taste. According to the preference test, the mixed-cultured radish juice incubated at 25$^{\circ}C$with Lactobacillus brevis and Leuconostoc mesenteroides were evaluated superior to commercial Dongchimi. As a result, taste and quality of radish juice was improved by addition of starters.

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Physiological and Ecological Characteristics of Indigenous Soybean Rhizobia Distributed in Korea -V. Effects of Co-inoculation of R. japonicum and A. lipoferum on the Effectiveness of Symbiotic Nitrogen Fixation with Soybean (우리 나라 토착대두근류균(土着大豆根瘤菌)의 분포상태(分布狀態)와 생리(生理) 및 생태학적(生態學的) 특성(特性) -제(第)V보(報) 대두근류균(大豆根瘤菌)과 협생질소고정균(協生窒素固定菌)과의 상호접종효과(相互接種效果))

  • Ryu, Jin-Chang;Suh, Jang-Sun;Lee, Sang-Kyu;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.3
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    • pp.307-315
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    • 1988
  • This experiment was conducted to find out the effects of fertilizer-N and co-inoculation of the Rhizobium japonicum and the Azospirillum lipoferum on nodulation, $N_2$-fixation, and growth of soybean under in situ conditions. The results obtained were summarized as follows: 1. The yield of soybean dry matter was significantly greater in the R. japonicum alone, and the mixed inoculation of R. japonicum and A. lipoferum than those of un-inoculation. But inoculum applied by different strains did not significant effect on plant growth. The effects of nitrogen applied on soybean dry matter were higher in the ammonium sulfate than potassium nitrate, and decreased with increasing rates of two forms of nitrogen applied regardless of nitrogen source. 2. Acetylene redution activity was more increased in a single inoculation of R. japonicum than those of the mixed inoculation of the R. japonicum and the A. lipoferum, in cases of Danyeup cultivar, regardless of the form of combined nitrogen used. 3. Nodule mass and total nitrogenase activity per plant showed the positively significant effect in the interrelationship between dry matter of soybean and some factors related to nitrogen fixation efficiency. 4. The highest symbiotic effect in Danyeup cultivar was obtained when a single R. japonicum 84 Dy-1 strain was inoculated and fertilized with 18 mM potassium nitrate.

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