• 제목/요약/키워드: Bacterial Inoculation

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Sterilization of Bacteria, Yeast, and Bacterial Endospores by Atmospheric-Pressure Cold Plasma using Helium and Oxygen

  • Lee Kye-Nam;Paek Kwang-Hyun;Ju Won-Tae;Lee Yeon-Hee
    • Journal of Microbiology
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    • 제44권3호
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    • pp.269-275
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    • 2006
  • Atmospheric-pressure cold plasma (APCP) using helium/oxygen was developed and tested as a suitable sterilization method in a clinical environment. The sterilizing effect of this method is not due to UV light, which is known to be the major sterilization factor of APCP, but instead results from the action of reactive oxygen radicals. Escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae deposited on a nitrocellulose filter membrane or Bacillus subtilis spores deposited on polypropylene plates were exposed to helium/oxygen plasma generated with AC input power at 10 kHz, 6 kV. After Plasma treatment, nitrocellulose filter membranes were overlaid on fresh solid media and CFUs were counted after incubation overnight. D-values were 18 sec for E. coli, 19 sec for S. aureus, 1 min 55 sec for S. cerevisiae, and 14 min for B. subtilis spores. D-values of bacteria and yeast were dependent on the initial inoculation concentration, while the D-value of B. subtilis spores showed no correlation. When treated cells were observed with a scanning electron microscope, E. coli was more heavily damaged than S. aureus, S. cevevisiae exhibited peeling, and B. subtilis spores exhibited shrunken morphology. Results showed that APCP using helium/oxygen has many advantages as a sterilization method, especially in a clinical environment with conditions such as stable temperature, unlimited sample size, and no harmful gas production.

Optimization of Indole-3-acetic Acid (IAA) Production by Bacillus megaterium BM5

  • Lee, Jae-Chan;Whang, Kyung-Sook
    • 한국토양비료학회지
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    • 제49권5호
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    • pp.461-468
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    • 2016
  • One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

Graft Transmission and Cytopathology of Pear Black Necrotic Leaf Spot (PBNLS) Disease

  • Nam, Ki-Woong;Kim, Kyung-Soo
    • The Plant Pathology Journal
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    • 제18권6호
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    • pp.301-307
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    • 2002
  • Graft transmission and cytopathological studies of a severe pear disease, pear black necrotic leafspot(PBNLS), were carried out to determine the causal agent of the disease. No evidence was found that a fungal or bacterial pathogen could be the causal agent of the disease. Attempts to transmit the agent by sap-inoculation to other plants including herbaceous hosts failed. How-ever, the pathogen was readily graft-transmitted from symptomatic diseased pears to healthy pears. Graft transmission of the pathogen was also demonstrated by using an indicator plant, PS-95, developed in the laboratory through various grafting methods. Ultrastructural study of the disease revealed the consistent presence of flexuous rod-shaped virus-like particles (VLP) in the symptomatic leaves of both Niitaka cultivar and indicator pear, PS-95. The particles, approximately 12 nm in diameter with undetermined length, occurred in the cytoplasm of mesophyll parenchyma cells. Cells with VLPs also contained fibril-containing vesicles, which are common in cells infected with plant viruses with ssRNA genome. The vesicles were formed at the tonoplast. Based on the symptomatology, the presence of fibril-containing vesicles, and graft-transmissibility, it is believed that the VLPs that occurred on symptomatic leaves of black necrotic leafspot of pear are viral in nature, possibly those of a capillovirus.

Ultrastructure of Initial Cytological Changes of Cowpea in Root Nodule Formation

  • Kim, Young-Ho;Cheon, Choong-ll
    • The Plant Pathology Journal
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    • 제15권2호
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    • pp.127-130
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    • 1999
  • Cytological changes of cowpea root at the early stage of root nodule formation (within 5 days after inoculation) were viewed by light and electron microscopy. The root region affected by the rhizobial infection, which was composed of a redial array of cortical cells, had prominent cell divisions, mostly anticlinal in the inner cortical cells and in addition oblique and periclinal in the outer cells. An infected root hair cell (or root hair-producing epidermal cell) had numerous infection threads and degenerated cytoplasm. Module meristem was formed adjacent to the infected root hair cell, and characterized by dense cytoplasm, prominent nucleus, numerous small vacuoles, and increased plastids, containing infection threads as well. Bacterial cells were dividing inside the infection thread, the wall materials of which appeared to be dissolved ad accumulated in small vacuoles. inner cortical cells contiguous to the nodule meristem appeared to be actively dividing and dedifferentiating; however, they were not infected by the rhizobia. These structural characteristics are similar to those in the Bradyrhizobium-soybean association previously reported, and may reflect the similar cytological process in cowpea in the early nodule formation.

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First Report of Melon Soft Rot Disease Caused by Pectobacterium brasiliense in Korea

  • Kyoung-Taek Park;Leonid N. Ten;Chang-Gi Back;Soo-Min Hong;Seung-Yeol Lee;Jeung-Sul Han;Hee-Young Jung
    • 식물병연구
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    • 제29권3호
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    • pp.310-315
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    • 2023
  • In May 2021, characteristic soft rot symptoms, including soft, watery, slimy, black rot, wilting, and leaf collapse, were observed on melon plants (Cucumis melo) in Gokseong, Jeollanam-do, Korea. A bacterial strain, designated KNUB-06-21, was isolated from infected plant samples, taxonomically classified, and phylogenetically analyzed using 16S rRNA and housekeeping gene sequencing. Strain KNUB-06-21 was also examined for compound utilization using the API ID 32 GN system and strain KNUB-06-21 was identified as Pectobacterium brasiliense. Subsequent melon stem inoculation studies using strain KNUB-06-21 showed soft rot symptoms similar to field plants. Re-isolated strains shared phenotypic and molecular characteristics with the original P. brasiliense KNUB-06-21 strain. To our knowledge, ours is the first report of P. brasiliense causing melon soft rot disease in Korea.

Pectobacterium brasiliense as a Causative Agent for Soft Rot of Radish in Korea

  • Kyoung-Taek Park;Soo-Min Hong;Chang-Gi Back;In-Kyu Kang;Seung-Yeol Lee;Leonid N. Ten;Hee-Young Jung
    • 식물병연구
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    • 제29권1호
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    • pp.64-71
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    • 2023
  • In October 2021, soft rot disease seriously affected radish crop in Dangjin, Chungcheongnam-do, Korea. The infected radishes were stunted and turned dark green, with yellowish leaf foliage. A slimy, wet, and decayed pith region was observed in the infected roots. The bacterial strain KNUB-03-21 was isolated from infected roots. The biochemical and morphological characteristics of the isolate were similar to those of Pectobacterium brasiliense. Phylogenetic analysis based on the sequences of the 16S rRNA region and the concatenated DNA polymerase III subunit tau (dnaX), leucine-tRNA ligase (leuS), and recombinase subunit A (recA) genes confirmed that the isolate is a novel strain of P. brasiliense. Artificial inoculation of radish with P. brasiliense KNUB-03-21 resulted in soft rot symptoms similar to those observed in infected radish in the field; subsequently, P. brasiliense KNUB-03-21 was reisolated and reidentified. To our knowledge, this is the first report of P. brasiliense as a causal pathogen of radish soft rot in Korea.

First Report of Pectobacterium versatile as the Causal Pathogen of Soft Rot in Kimchi Cabbage in Korea

  • Kyoung-Taek Park;Soo-Min Hong;Chang-Gi Back;Young-Je Cho;Seung-Yeol Lee;Leonid N. Ten;Hee-Young Jung
    • 식물병연구
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    • 제29권1호
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    • pp.72-78
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    • 2023
  • In September 2021, gray-to-brown discoloration and expanding water-soaked lesions were observed on the outer and inner layers and the core of kimchi cabbage (Brassica rapa subsp. pekinensis) in fields located in Samcheok, Gangwondo, Korea. A bacterial strain designated as KNUB-02-21 was isolated from infected cabbage samples. Phylogenetic analysis based on the sequences of the 16S rRNA region and the dnaX, leuS, and recA genes confirmed that the strain was affiliated with Pectobacterium versatile. Additionally, the biochemical and morphological profiles of the isolate were similar to those of P. versatile. Based on these results, the isolate was identified as a novel strain of P. versatile. Healthy kimchi cabbage slices developed soft rot upon inoculation with P. versatile KNUB-02-21 and exhibited symptoms similar to those observed in the diseased plants in fields. The re-isolated strains were similar to those of P. versatile. Prior to our study, P. versatile as the causative pathogen of kimchi cabbage soft rot had not been reported in Korea.

First Report of Pectobacterium brasiliense Causing Bitter Melon Soft Rot Disease in Korea

  • Kyoung-Taek Park;Leonid N. Ten;Soo-Min Hong;Chang-Gi Back;Seung-Yeol Lee;Hee-Young Jung
    • 식물병연구
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    • 제29권4호
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    • pp.452-458
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    • 2023
  • In the Goesan region, located in Chungcheongbuk-do, Korea, a significant outbreak of soft rot infections was documented in August 2021, affecting fruits of Momordica charantia, commonly known as bitter melon or bitter gourd. The symptoms included a noticeable transition to yellowing in the affected fruits, eventually leading to their collapse. The bacterial strain KNUB-09-21 was isolated from the diseased fruits. Molecular analysis, using the sequences of the 16S rRNA region and three housekeeping genes (dnaX, recA, and leuS), along with the results of compound utilization in the API ID 32 GN system, provide strong evidence for the identification of the isolate KNUB-09-21 as Pectobacterium brasiliense. The pathogenicity of strain KNUB-09-21 on M. charantia was confirmed through a controlled inoculation test. Within two days, inoculated fruits displayed soft rot symptoms closely resembling those observed in naturally affected fruits. This is the first report of soft rot on M. charantia in Korea.

Identification and genetic characterization of bacterial isolates causing brown blotch on cultivated mushrooms in Korea

  • Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
    • 한국버섯학회지
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    • 제22권2호
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    • pp.37-47
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    • 2024
  • Fluorescent bacteria were isolated from sporocarps that browned into various mushrooms during survey at places of the production in Korea. We examined the pathogenicity, biodiversity, and genetic characteristics of the 19 strains identified as Pseudomonas tolaasii by sequence analysis of 16S rRNA and White Line Assay. The results emphasize the importance of rpoB gene system, fatty acid profiles, specific and sensitive PCR assays, and lipopeptide detection for the identification of P. tolaasii. As a result of these various analyses, 17 strains (CHM03~CHM19) were identified as P. tolaasii. The phylogenetic analysis based on the 16S rRNA gene showed that all strains were clustered closest to P. tolaasii lineage, two strains (CHM01, CHM02) were not identified as P. tolaasii and have completely different genetic characteristics as a result of fatty acids profile, specific and sensitive PCR, lipopetide detection, rpoB sequence and REP-PCR analysis. Pathogenicity tests showed 17 strains produce severe brown discolouration symptoms to button mushrooms and watersoaking of sporophore tissue within three days after inoculation. But two strains did not produce discolouration symptoms. Therefore, these two strains will be further investigated for correct species identification by different biological and molecular characteristics.

Bacteriophage의 Salmonella enterica serovar Enteritidis에 대한 in vitro 및 in vivo 효능 평가 (Evaluation of in vitro and in vivo bacteriophage efficacy against Salmonella enterica serovar Enteritidis infection)

  • 차승빈;;이원정;신민경;노유미;정명환;명길선;안영태;허철성;유한상
    • 대한수의학회지
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    • 제50권3호
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    • pp.213-220
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    • 2010
  • Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.