• Title/Summary/Keyword: Bacterial Inoculation

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

  • Cha, Seung-Bin;Rayamajhi, Nabin;Lee, Won-Jung;Shin, Min-Kyoung;Roh, Yu-Mi;Jung, Myung-Hwan;Myoung, Kil-Sun;Ahn, Young-Tae;Huh, Chul-Sung;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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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.

Control Efficacy of Fungicides on Pepper Bacterial Wilt (고추 풋마름병에 대한 살균제의 방제 효과)

  • Lee, Soo Min;Kwak, Yeon Soo;Lee, Kyeong Hee;Kim, Heung Tae
    • The Korean Journal of Pesticide Science
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    • v.19 no.3
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    • pp.323-328
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    • 2015
  • Control efficacy was investigated with fungicides as 3 copper compound, 3 antibiotic fungicides and one fungicide containing to quinolone against the growth of Ralstonia solanacearum on NA medium and the disease occurrence on pepper seedlings. Among 7 fungicides, oxytetracycline was shown the highest activity against a growth of the pathogen in the agar diffusion method, but validamycin showed no activity against the pathogen. With $1000{\mu}g\;mL^{-1}$ of each copper fungicide as copper hydroxide, copper oxychloride+ dithianone and copper sulfate, 2.2, 1.3 and 1.5 mm in size of clear zone only could be found, respectively. In pepper seedling test, oxytetracycline showed a perfect activity in all treatments 7 days after inoculation. However, its activity was decreased from $500{\mu}g\;mL^{-1}$ of treatment over the time. Copper fungicides showed the control efficacy lower than antibiotic fungicides except for validamycin. Based on the results, it was suggested that it would be better to use antibiotic fungicides than copper fungicides to control pepper bacterial wilt in the fields.

Antibacterial Activity of Pharbitin, Isolated from the Seeds of Pharbitis nil, against Various Plant Pathogenic Bacteria

  • Nguyen, Hoa Thi;Yu, Nan Hee;Park, Ae Ran;Park, Hae Woong;Kim, In Seon;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1763-1772
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    • 2017
  • This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibition concentration (MIC) value of $250{\mu}g/ml$. Among the three solvent layers obtained from the methanol extract of P. nil seeds, only the butanol layer displayed the activity with an MIC value of $125{\mu}g/ml$ against Xap. An antibacterial fraction was obtained from P. nil seeds by repeated column chromatography and identified as pharbitin, a crude resin glycoside, by instrumental analysis. The antibacterial activity of pharbitin was tested in vitro against 14 phytopathogenic bacteria, and it was found to inhibit Ralstonia solanacearum and four Xanthomonas species. The minimum inhibitory concentration values against the five bacteria were $125-500{\mu}g/ml$ for the n-butanol layer and $31.25-125{\mu}g/ml$ for pharbitin. In a detached peach leaf assay, it effectively suppressed the development of bacterial leaf spot, with a control value of 87.5% at $500{\mu}g/ml$. In addition, pharbitin strongly reduced the development of bacterial wilt on tomato seedlings by 97.4% at $250{\mu}g/ml$, 7 days after inoculation. These findings suggest that the crude extract of P. nil seeds can be used as an alternative biopesticide for the control of plant diseases caused by R. solanacearum and Xanthomonas spp. This is the first report on the antibacterial activity of pharbitin against phytopathogenic bacteria.

Effective Components of Commercial Enzyme Food Products and Their HACCP Scheme (시판 효소식품의 유용성분과 HACCP 관리 방안에 관한 연구)

  • Lee, Eun-Joo;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.33 no.4
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    • pp.461-468
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    • 2001
  • The effectiveness and safety of Enzyme Food, a group of dietary supplements designated by Korean Food Law, were evaluated and the possibility of HACCP (Hazard Analysis Critical Control Point) application was investigated. Chemical composition, enzyme activities and the degree of bacterial contamination in 12 samples of different brands sold in Korean market were measured. The chemical composition of the selected products varied and inconsistent to those claimed in the label description. It is known that effectiveness of Enzyme Food depends on enzyme activity, but enzyme activities of ${\alpha}-amylase$ varied from $1,793\;{\mu}g/min$ g to $159\;{\mu}g/min$ g and those of ${\beta}-amylase$ ranged from $171\;{\mu}g/min$ g to $11\;{\mu}g/min$ g. The protease activities varied from $27.57\;{\mu}g/min$ g to $0.18\;{\mu}g/min$ g. In coli-form bacterial test, positive reactions were appeared in the 50 % of the samples. Numbers of bacteria ranged from $1.3\;{\times}\;10^5\;to\;1.2\;{\times}\;10^9$. Five CCPs were identified; heating, inoculation, cultivation, drying and granulation. Consideration of HACCP system indicated that the pretreatment of raw material, checking of bacterial contamination and stability of enzyme activity during fermentation process were important factors for the quality of Enzyme Foods.

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Isolation and Characterization of an Antagonistic Endophytic Bacterium Bacillus velezensis CB3 the Control of Citrus Green Mold Pathogen Penicillium digitatum (감귤저장병 병원균 Penicillium digitatum 방제를 위한 길항 내생세균 Bacillus velezensis CB3의 분리 및 특성 규명)

  • Lee, Ji-Hyun;Seo, Mun-Won;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.40 no.2
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    • pp.118-123
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    • 2012
  • In order to develop environment friendly fungicide for the control of citrus green mold (Penicillium digitatum) using endophytic bacteria, the 21 bacterial isolates were isolated from citrus leaves in seven different orchards in Jeju Province. Among the 21 bacterial isolates, 5 bacterial isolates presented antifungal activity against green mold pathogen P. digitatum. The CB3 isolate, which showed the most strong antagonistic effect, was selected through opposite culture against the pathogen. The rod-shaped, gram-positive bacterium CB3 was identified as Bacillus velezensis based on morphological, physiological characteristics, 16S rDNA, and gyr A gene sequence analysis. The isolate CB3 showed strong antifungal activity against two citrus postharvest pathogen P. digitatum. Citrus fruits were treated by wound inoculation with P. digitatum pathogen, and the control efficacy of CB3 culture broth was 66.7% ($1{\times}10^8$ cfu/ml). In conclusion, The stability of CB3 and its strong antifungal activity also lead us to believe that it has potential for application as an environment friendly biological control agent.

Effects of N,P,K fertilizer levels and growth condition on the development of Bacterial leaf blight in rice plants (삼요소시비량과 수도생육상태가 백엽고병(벼, 흰빛잎마름병) 발병에 미치는 영향)

  • Kim Chung Hwa;Cho Young Sup
    • Korean journal of applied entomology
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    • v.9 no.1
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    • pp.7-13
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    • 1970
  • 1. Kum Nam Poong which is highly susceptible to bacterial leaf blight was used as the host plant throughout this experiment. Xanthomonas oryzae 6526 was inoculated on the top of upper leaves by single needle inoculation method. After 14 days, the enlarged spots were examined in the experimental pots. Each of 3 levels of nitrogen, phosphate and potassium pots were arranged at random with three replications. 2. The amounts of nitrogen fertilizer applied and the lesion development of bacterial leaf blight were positively correlated regardless of application of phosphate and potassium fertilizers. 3. The effect of phosphate fertilizer on the lesion development was not significantly different from standard level. The lesion development was stimulated when the amount of phosphate fertilizer was increased as twice as standard level. 4. The inhibitory effect of potassium fertilizer on leaf blight was maximum by applying standard level. The stimulative effect of potassium fertilizer on the lesion development, however, was noticed. when potassium fertilizer applied was increased as twice as the standard level. 5. The heading date and spike number of rice plant were significantly correlated with the lesion development, and such phenomena were depended on the amount of nitrogen fertilizer applied regardless of the other fertilizers applied in this experiment.

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Development of Molecular Markers Conferring Bacterial Leaf Pustule Resistance Gene, rxp, using Resistant and Susceptible Cultivars in Soybean (콩 불마름병 저항성 및 감수성 품종을 이용한 rxp 유전자 근접 분자표지 개발)

  • Yang, Kiwoung;Lee, Yeong Hoon;Ko, Jong Min;Jeon, Myeong Gi;Lee, Byong Won;Kim, Hyun Tae;Yun, Hong Tae;Jung, Chan Sik;Baek, In Youl
    • Korean Journal of Breeding Science
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    • v.43 no.4
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    • pp.282-287
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    • 2011
  • Bacterial pustule (BP) is a leaf disease of soybean that is most common in Korea. Inoculation of 8ra, pathogen strain, to resistant and susceptible cultivars for finding the BP resistance gene (rxp) was much tried but the sequence of the exact gene is not found. This research performed in order to confirm the rxp gene near molecular marker by using the resistant and susceptible cultivars. Soybean BP resistance gene which related to region of near molecular marker could select the resistant cultivar. For the near molecular marker of rxp, reference genomics data available at sequenced Phytozome was used for designing molecular markers. The rxp was mapped between Satt372 and Satt486 on chromosome 17. According to previous study, rxp released in find mapping 7.2 Mbp to 7.3 Mbp on chromosome 17. In this study, we developed 3 random markers near from 6.6 Mbp to 7.3 Mbp on chromosome 17 identified to increase the genetic resolution of the rxp gene region using resistant and susceptible cultivars. Particularly, Rxp17-700 marker was mostly coincided resistance and susceptible genotype to rxp. This result suggests that Rxp17-700 marker will be more tightly linked to rxp gene.

Generation of Bacterial Blight Resistance Rice with Transcription Factor OsNAC69-overexpressing (전사인자 OsNAC69-과발현을 통한 흰잎마름병 저항성 벼 제작)

  • Park, Sang Ryeol;Cha, Eun-Mi;Moon, Seok Jun;Shin, Dongjin;Hwang, Duk-Ju;Ahn, Il-Pyung;Bae, Shin-Chul
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.457-463
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    • 2011
  • Plant specific gene family, NAC (NAM, ATAF, and CUC) transcription factors have been characterized for their roles in plant growth, development, and stress tolerance. In this study, we isolated OsNAC69 gene and analyzed expression level by inoculation of bacterial leaf blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo). NAC transcription factor family can be divided into five groups (I-V). On the basis of phylogenetic analysis, OsNAC69 was fall into group II. OsNAC69 was strongly induced 1 hr after infected with Xoo. To investigate its biological function in the rice, we constructed vector for overexpression in rice, and then generated transgenic rice lines. Gene expression of OsNAC69-overexpressed transgenic rice lines were analyzed by northern blot. Analysis of disease resistance to pathogen Xoo, nine OsNAC69-overexpressed transgenic rice lines showing high expression level of OsNAC69 were shown more resistant than wild type. These results suggest that OsNAC69 gene may play regulatory role during pathogen infection.

First Report of Bacterial Root Rot Caused by Serratia plymuthica on Panax ginseng (Serratia plymuthica에 의한 인삼 세균뿌리썩음병 발생)

  • Jung, Won Kwon;Kim, Young Soo;Choi, Jin Kook;Kim, Seung-Han;Jang, Myeong-Hwan;Kwon, Tae Lyong;Jeon, Yongho
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.332-338
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    • 2018
  • In August of 2011, a wilting disease of ginseng was observed at Bongwha, Gyeongbuk province, Korea. Affected plants initially show withering symptoms on leaves of ginseng. As the disease progresses, withering leaves spread downward, eventually encompassing the whole plant. Leaves lose vigor but remain pale green. Symptoms of roots were brown, and soft rots characterized by moist and watery decay of the whole ginseng root, which initiated as small brown, water-soaked lesions of hairy roots and enlarged to the entire roots. The causal organism isolated from the infected roots was identified as Serratia plymuthica based on its physiological and biochemical characteristics, by cellular fatty acid composition (GC-FAME), the utilization of carbon sources (BioLog System), and 16S rRNA sequence of the isolated bacterium were 99% homologous to those of Serratia plymuthica strains. Artificial inoculation of the bacterium produced the same brown or soft rot symptoms on the ginseng roots, from which the same bacterium was isolated. This is the first report of bacterial root rot caused by the Serratia plymuthica in ginseng in Korea. Serratia plymuthica has been used as antagonistic microorganism for biological control on several crop plants. But it was proved pathogen of ginseng at humid condition in this study.

Comparison of the amount of bacteria according to the number of holes between bristles (칫솔모 사이에 형성한 구멍 수에 따른 세균 양 변화)

  • Kang, Kyung-Hee;Kang, So-Hyeon;Kim, So-Hee;Kim, Ji-Ho;Baek, Su-Jeong;Seo, Hyeon-Ji;Yun, Hae-Yeon;Goong, Hwa-Soo
    • Journal of Korean Academy of Dental Administration
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    • v.6 no.1
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    • pp.19-27
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    • 2018
  • The purpose of this study was to determine the level of toothbrush bacterial growth, whether the dryness of the toothbrush head differs depending on the number of holes in the head, and to use these results as a reference for future toothbrush design. Two-millimeter holes were created on the head of the toothbrushes in groups of three, one, or zero holes. We made the solution with Streptococcus mutans, and the toothbrushes were placed in the solution and agitated. The toothbrushes were shaken to remove moisture and allowed to air-dry. The toothbrush heads were swabbed with saline and then placed in two inoculation groups. The first group was inoculated with a 102 dilution of the S. mutans culture and the second was inoculated with the original culture. After incubation, bacterial colony numbers were measured. The number of holes on the toothbrush head correlated with a decrease in number of cultured bacterial colonies. Our model of a toothbrush head with holes indicated that these holes in the toothbrush head were effective in reducing the level of microbial contamination and that a greater number of holes creates an improved toothbrush sanitation effect. The average number of colonies on the head of toothbrush by number of holes was high, followed by the number of holes 0, 1 and 3, and the average number of colony among toothbrush heads was same. The use of a toothbrush with holes between the toothbrush head indicates that it is effective in reducing the level of microbial contamination between the toothbrush head and toothbrush and the higher the number of holes, the better the effect.