• Title/Summary/Keyword: Pathogen Concentration

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Efficient Screening Method for Resistance of Cucumber Cultivars to Fusarium oxysporum f. sp. cucumerinum (오이 덩굴쪼김병에 대한 효율적인 저항성 검정 방법)

  • Lee, Ji Hyun;Kim, Jin-Cheol;Jang, Kyoung Soo;Choi, Yong Ho;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.20 no.4
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    • pp.245-252
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    • 2014
  • The study was performed to establish an efficient screening method for resistant cucumber to Fusarium oxysporum f. sp. cucumerinum. The isolate KR5 was identified as F. oxysporum f. sp. cucumerinum based on molecular analyses of ITS and TEF genes and host-specificity test on cucurbits including melon, oriental melon, cucumber, and watermelon. Then four cucumber and two rootstock cultivars showing different resistance degrees to the Fusarium wilt pathogen KR5 were selected. And development of Fusarium wilt of the six cultivars according to several conditions, including incubation temperature after inoculation, inoculum concentration, root wounding, and growth stages of seedlings, was investigated. Disease severity of Fusarium wilt on the resistant cultivars was changed with incubation temperatures after inoculation. The resistant cultivars showed the higher resistance when inoculated plants were kept at 25 or $30^{\circ}C$ than at $20^{\circ}C$. Among four different growth stages of the seedlings, seven-day-old seedling represented the most difference of resistance and susceptibility to Fusarium wilt. From above results, we suggest that an efficient screening method for resistant cucumber to F. oxysporum f. sp. cucumerinum is to dip the non-cut roots of seven-day-old seedlings in spore suspension of $1.0{\times}10^6-1.0{\times}10^7$ conidia/ml and to transplant the seedling into a non-infected soil, and then to incubate the inoculated plants in a growth room at $25^{\circ}C$ for 3 weeks to develop Fusarium wilt.

Inhibitory Effect of Transition Metal Gallium [Ga(NO3)3] on Biofilm Formation by Fish Pathogens (전이금속 갈륨(Ga(NO3)3)을 이용한 biofilm을 형성하는 어류질병세균의 억제)

  • Kim, Dong-Hwi;Dharaneedharan, Subramanian;Jang, Young-Hwan;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.535-539
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    • 2016
  • The prevalence of pathogenic bacteria such as Streptococcus parauberis (Sp), Streptococcus iniae (Si), and Edwardsiella tarda (Et) in flounder fish farms in Jeju Island and their management by gallium treatment was studied. Sp, Si, and Et were found to exhibit a low rate of cell growth and high biofilm formation. Hence, in the present study, cell growth and biofilm formation were measured spectrophotometrically 72 h after the addition of different concentrations of gallium (2, 4, or 8 mg/ml). In addition, cell death was measured by resazurin and propidium iodide staining assays. The results showed that bacterial cell death increased and biofilm formation decreased with an increasing concentration of gallium. Hence, the present study signifies that the use of gallium against bacterial pathogens could be useful for disease management in flounder farms.

Analysis of Bacterial Spot Disease in Red Pepper Caused by Increase of CO2 Concentration (CO2 농도 상승 효과에 의한 고추 세균점무늬병 발병 양상 분석)

  • Jang, Jong-Ok;Kim, Byung-Hyuk;Moon, Doo-Gyung;Koh, Sang-wook;Joa, Jae-Ho
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.77-84
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    • 2018
  • An increase in $CO_2$ will affect plant pathogenic microorganisms, the resistance of host plants, and host-pathogen interactions. This study used Capsicum annuum and Xanthomonas euvesicatoria, a pathogenic bacterium of pepper, to investigate the interactions between hosts and pathogens in conditions of increased $CO_2$ concentrations. Our analysis of disease resistance genes under 800 ppm $CO_2$ using quantitative RT-PCR showed that the expression of CaLRR1, CaPIK1, and PR10 decreased, but that of negative regulator WRKY1 increased. Additionally, the disease progress and severity was higher at 800 ppm than 400 ppm $CO_2$. These results will aid in understanding the interaction between red pepper and X. euvesicatoria under increased $CO_2$ concentrations in the future.

Diversity of Endophytic Fungi from Different Verticillium-Wilt-Resistant Gossypium hirsutum and Evaluation of Antifungal Activity Against Verticillium dahliae In Vitro

  • Li, Zhi-Fang;Wang, Ling-Fei;Feng, Zi-Li;Zhao, Li-Hong;Shi, Yong-Qiang;Zhu, He-Qin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1149-1161
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    • 2014
  • Cotton plants were sampled and ranked according to their resistance to Verticillium wilt. In total, 642 endophytic fungi isolates representing 27 genera were recovered from Gossypium hirsutum root, stem, and leaf tissues, but were not uniformly distributed. More endophytic fungi appeared in the leaf (391) compared with the root (140) and stem (111) sections. However, no significant difference in the abundance of isolated endophytes was found among resistant cotton varieties. Alternaria exhibited the highest colonization frequency (7.9%), followed by Acremonium (6.6%) and Penicillium (4.8%). Unlike tolerant varieties, resistant and susceptible ones had similar endophytic fungal population compositions. In three Verticillium-wilt-resistant cotton varieties, fungal endophytes from the genus Alternaria were most frequently isolated, followed by Gibberella and Penicillium. The maximum concentration of dominant endophytic fungi was observed in leaf tissues (0.1797). The evenness of stem tissue endophytic communities (0.702) was comparatively more uniform than the other two tissues. Eighty endophytic fungi selected from 27 genera were evaluated for their inhibition activity against highly virulent Verticillium dahliae isolate Vd080 in vitro. Thirty-nine isolates exhibited fungistasis against the pathogen at varying degrees. Seven species, having high growth inhibition rates (${\geq}75%$), exhibited strong antifungal activity against V. dahliae. The antifungal activity of both volatile and nonvolatile metabolites was also investigated. The nonvolatile substances produced by CEF-818 (Penicillium simplicissimum), CEF-325 (Fusarium solani), CEF-714 (Leptosphaeria sp.), and CEF-642 (Talaromyces flavus) completely inhibited V. dahliae growth. These findings deepen our understanding of cotton-endophyte interactions and provide a platform for screening G. hirsutum endophytes with biocontrol potential.

Inhibition of Microbial Quorum Sensing Mediated Virulence Factors by Pestalotiopsis sydowiana

  • Parasuraman, Paramanantham;Devadatha, B;Sarma, V. Venkateswara;Ranganathan, Sampathkumar;Ampasala, Dinakara Rao;Reddy, Dhanasekhar;Kumavath, Ranjith;Kim, In-Won;Patel, Sanjay K.S.;Kalia, Vipin Chandra;Lee, Jung-Kul;Siddhardha, Busi
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.571-582
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    • 2020
  • Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.

Study on the Removal of Pharmaceuticals and Personal Care Products and Microorganism Inactivation by Ozonation (오존처리에 의한 의약품류의 제거와 미생물의 불활성화에 대한 연구 및 고찰)

  • Kim, Il-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.12
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    • pp.1134-1140
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    • 2010
  • Ozonation is a promising process that can effectively reduce the occurrence of micropollutants and pathogen in water. This study investigated the performance of ozonation for the removal of pharmaceuticals and personal care products (PPCPs) in secondary effluent from wastewater treatment plant. Moreover, the disinfection potential of ozonation applied for PPCPs removal was discussed. Secondary effluent filtered by sand filter was used for tested water, and ozonation was performed under 2, 4 and 6 mg/L of ozone doses. As a result, 6 mg/L of ozone dose (ozone consumption : 4.4 mg/L) was essential for the effective removal of 37 PPCPs in tested water. Several previous studies showed that the operation condition could achieve approximately 3 log inactivation of total coliform and enteroviruses. On the other hand, dissolved ozone concentration in tested water increased by 1.8 mg/L under 6 mg/L of ozone dose, probably resulting in the increase of bromate formation potential. This result implies that as alternatives to suppress the bromate formation potential during the oxidation of PPCPs by ozone, investigations on advanced oxidation processes are required.

Characterization of the Nickel Resistance Gene from Legionella pneumophila: Attenuation of Nickel Resistance by ppk (polyphosphate kinase) Disruption in Escherichia coli

  • Hahm, Dae-Hyun;Yeon, Mi-Jung;Ko, Whae-Min;Lee, Eun-Jooh;Lee, Hye-Jung;Shim, In-Sop;Kim, Hong-Yeoul
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.114-120
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    • 2002
  • A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2+$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.

Effects of various concentrations of skullcap extract in the diets on disease resistance of olive flounder, Paralichthys olivaceus (생약재 황금 뿌리 열수추출물의 넙치 투여시 질병저항성에 미치는 영향)

  • Jee, Bo-Young;Seo, Jung-Soo;Jeon, Eun-Ji;Lee, Eun-Hye;Choi, Hee-Jung;Kim, Jin-Do;Jung, Sung-Hee;Park, Myoung-Ae
    • Journal of fish pathology
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    • v.25 no.1
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    • pp.21-30
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    • 2012
  • Effects of various concentration of skullcap Scutellaria baicalensis in the diets on a nonspecific immunity and a disease resistance of olive flounder were investigated. After feeding trial, weight gain of fish fed 0.05% skullcap immersed group was higher than that of fish fed 0, 0.1 and 1% skullcap diet but no significant differences were observed among the experimental groups. Furthermore, no significant differences in hematological indices of olive flounder were found among the experimental groups. Lysozyme activity in the serum and kidney of the administrated group(0.05% skullcap immersed group) was significantly higher than the control group. In addition, the chemiluminescent(CL) responses of head kidney leucocytes from the 0.05% skullcap immersed group was significantly higher(P<0.05) than the control group. In the histological results, the 1% skullcap immersed group appeared to have the detrimental effects for fish health. In a challenge experiment with Edwardsiella tarda(GY-01) and Streptococcus iniae(FT5228), relative percent survival (RPS) in the 0.05% skullcap immersed group was higher than the control group injected with E. tarda(GY-01) at $4^{th}$ and $8^{th}$ weeks. The results suggest that the skullcap extract (0.05%) would be effective to enhance the nonspecific immunity and protective ability of olive flounder against fish pathogen such as E. tarda.

Development of Convenient Screening Method for Resistant Radish to Plasmodiophora brassicae (효율적인 무 뿌리혹병 저항성 검정법 확립)

  • Jo, Su-Jung;Jang, Kyoung-Soo;Choi, Yong-Ho;Kim, Jin-Cheol;Choi, Gyung-Ja
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.161-168
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    • 2011
  • To establish simple and reliable screening method for resistant radish to Plasmodiophora brassicae Woron. using soil-drenching inoculation, the development of clubroot on radish seedlings inoculated with P. brassicae GN-1 isolate according to several conditions such as inoculum concentration, plant growth stage and incubation period after inoculation was studied. To select resistant radish against clubroot, 10-day-old seedlings were inoculated with P. brassicae by drenching the roots with the spore suspension of the pathogen to give $1{\times}10^9$ spores/pot. The inoculated seedlings were incubated in a growth chamber at $20^{\circ}C$ for 3 days then cultivated in a greenhouse ($20{\pm}5^{\circ}C$) for 6 weeks. Under the optimum conditions, 46 commercial cultivars of radish were tested for resistance to YC-1 (infecting 15 clubroot-resistant cultivars of Chinese cabbage) and GN-1 (wild type) isolates of P. brassicae. Among them, thirty-five cultivars showed resistance to both isolates and one cultivar represented susceptible response to the pathogens. On the other hand, the other cultivars showed different responses against the tested P. brassicae pathogens. The results suggest that this method is an efficient system for screening radish with resistance to clubroot.

Effects of propolis extracts on the immune response in cultured flounder, Paralichthys olivaceus (Propolis extracts가 양식넙치의 면역활성능에 미치는 영향)

  • Nam, Hyun Ju;Park, Kyung Il;Choi, Min Soon
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.47-56
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    • 2014
  • Propolis is a beehive product with a very complex chemical composition, widely used in folk medicine because of its several therapeutic activities. This study was conducted to measure the efficacy of propolis on non-specific defense reactions, specific immune response, and protection levels against pathogen challenge with Streptococcus iniae. in vitro and in vivo. In vitro, the phagocytic activity and NBT assay of peripheral blood leukocytes (PBL) were evaluated in a various propolis extractsconcentrations (0, 10, 50, 100, 150, 250 and $500{\mu}g/ml$). The optimal concentration showing activation of propolis extracts was determined to $100{\mu}g/ml$. In vivo, they were divided into four groups (PBS, propoli extractss, vaccine, propolis extracts + vaccine) in vivo. Fish were received i.p. injection of either PBS or propolis extracts, and in the presence or absence of formalin inactivated S. iniae ($1{\times}10^8$ CFU/fish), respectively. The level of haematocrit is not affected among experimental groups. The phagocytic activity and the NBT reduction activities of head kidney phagocyte were markedly (p<0.05)augmented in the propolis extracts groups than in the PBS-control group, respectively. The level of serum lysozyme activity was significantly (p<0.05) increased in the propolis extracts treated groups than in the PBS-control group. The agglutinin titer was significantly (p<0.05) enhanced in the vaccine+propolis extracts group than in the vaccine group, but there was no difference between PBS-control and propolis treated group. The results of the present study suggest that propolis extracts seems to be a promising compounds of non-specific immune stimulator, also being able to use a good adjuvant.