• 제목/요약/키워드: environmental DNA (eDNA)

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토양에 매몰 방제된 화상병 감염 사과와 배 나무로부터 화상병균 생존 조사 (Investigating Survival of Erwinia amylovora from Fire Blight-Diseased Apple and Pear Trees Buried in Soil as Control Measure)

  • 김예은;김준영;노형진;이동형;김수산;김성환
    • 한국환경농학회지
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    • 제38권4호
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    • pp.269-272
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    • 2019
  • BACKGROUND: Since 2015, fire blight disease caused by Erwinia amylovora has been devastating apple and pear orchards every year. To quickly block the disease spreading, infected apple and pear trees have been buried in soil. However, concern on the possibility of the pathogen survival urgently requires informative data on the buried host plants. Therefore, this study was conducted to investigate the survival of the pathogen from the buried host plants. METHODS AND RESULTS: Apple trees buried in 42 months ago in a Jecheon site and pear trees buried in 30 months ago in an Anseong site were excavated using an excavator. Plant samples were taken from stems and twigs of the excavated trees. The collected 120 samples were checked for rotting and used for bacterial isolation, using TSA, R2A, and E. amylovora selection media. The purely isolated bacteria were identified based on colony morphology and 16S rDNA sequences. Wood rotting and decay with off smells and discoloring were observed from the samples. A total of 17 genera and 48 species of bacteria were identified but E. amylovora was not detected. CONCLUSION: Our investigation suggests that the survival of E. amylovora doesn't seem possible in the infected hosts which have been buried in soil for at least 30 months. Therefore, the burial control can be considered as a safe method for fire blight disease.

나린진(Naringin)의 $CCl_4$에 의한 급성 간독성 보호효과 (Protective Effect Naringin on Carbon Tetrachloride Induced Hepatic Injury in Mice)

  • 채수철;고은경;최승현;유근창
    • Environmental Analysis Health and Toxicology
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    • 제23권4호
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    • pp.325-335
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    • 2008
  • The protective effects of the Naringin, on carbon tetrachloride ($CCl_4$)-induced hepatotoxicity and the possible mechanisms involved in this protection were investigated in mice. Pretreatment with Naringin prior to the administration of $CCl_4$ significantly prevented an increase in serum alanine, aspartate aminotransferase activity and hepatic lipid peroxidation in a dose-dependent manner. In addition, pretreatment with Naringin also significantly prevented the depletion of glutathione (GSH) content in the livers of $CCl_4$-induced mice. However, reduced hepatic glutathione levels was unaffected by treatment with Naringin alone. In addition, Naringin prevented $CCl_4$-induced apoptosis and necrosis, as indicated by a liver DNA laddering. To determine whether caspase-8,-3 pathway involved in $CCl_4$-induced acute liver injury, caspase-8, -3 activities were tested by ELISA. Naringin attenuated $CCl_4$induced caspase-8, -3 activities in mouse livers. $CCl_4$-induced hepatotoxicity was also prevented, as indicated by a liver histopathologic study. The effects of Naringin on the cytochrome P450 (CYP) 2E1, the major isozyme involved in $CCl_4$ were also investigated. Treatment of mice with Naringin resulted in a significant decrease of the CYP2E1-dependent hydroxyl at ion and aniline in a dose-dependent manner. These findings suggest that protective effects of Naringin against the $CCl_4$-induced hepatotoxicity may be due to its ability to block CYP2E1-mediated $CCl_4$ bioactivation and that is also protects against caspase-8, -3 pathway mediated apoptosis.

지표미생물을 이용한 시화호 유입수의 수질평가 (Evaluation of Influent Water Quality Using Indicator Microorganisms in Lake Shiwha)

  • 이희태;김희연;박현진;조영은;유소영;이경진;정종선;고광표
    • 한국환경보건학회지
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    • 제34권1호
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    • pp.86-94
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    • 2008
  • Lake Shiwha, an artificial lake located near metropolitan Seoul, offers a unique water environment and has been suspected to have high levels of chemical and microbiological contaminations. Lake Shiwha was originally connected to the sea but currently has four major surface water inputs from agricultural, municipal, industrial areas and in addition an occasional inflow from the sea. The objectives of this study are to investigate the relative contribution of microbial contaminants from each of the inflowing surface waters and to identify appropriate microbial indicator organisms in this unique water environment. We measured the levels of microbial contaminations in the four inflowing surface waters. A number of microbial indicator organisms including total coliform (TC), fecal coliform (FC), E. coli, Enterococci, somatic and male-specific coliphages were analyzed. Bacterial indicator microorganisms were detected and quantified by the $Colilert^{(R)},\;Enterolert^{(R)}$ kit. Surface water (50 l) was sampled by $ViroCap^{TM}\;5"$ cartridge filters and analyzed by the single agar layer method for detecting coliphages. The concentrations of TC, FC, E. coli, and Enterococci were 1543 CFU/100 ml${\sim}1.99{\times}10^6$ CFU/100 ml, 0 CFU/100 ml${\sim}202$ CFU/100ml, 0 CFU/100 ml${\sim}1.80{\sim}10^5$ CFU/100ml, 74 CFU/100 ml${\sim}3408$ CFU/100 ml, respectively. The male-specific and somatic coliphages were detected in three different inflowing surface waters. Isolated E. coli and Enterococci strains were further analyzed by 16s rDNA amplification and subsequent phylogenetic analysis from Jungwang-chun, Ansan-chun, Banwol-chun and penstock of inflowing surface water. Our results indicated that the concentrations of different fecal indicator microorganisms might not be highly correlated with each other. Multiple microbial indicator organisms should be used for monitoring microbial contamination and microbial source tracking methods.

Removal Behavior of Biological Nitrogen and Phosphorus and Prediction of Microbial Community Composition with Its Function, in an Anaerobic-Anoxic System form Weak Sewage

  • LEE, JIN WOO;EUI SO CHOI;KYUNG IK GIL;HAN WOONG LEE;SANG HYON LEE;SOO YOOUN LEE;YONG KEUN PARK
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.994-1001
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    • 2001
  • An easier way of understanding the BNR system was proposed from the study on substrate, nutrient removal tendency, microbial community and its metabolic function by applying the municipal settled sewage. During the anaerobic period, the phosphorus release rate per VFACOD we varied depending on the phosphorus content in the sludge. When the phosphorus content in the sludge was $6\%$ VSS, according to influent VFACOD, the phosphorus release rate and PHA production were $0.35 gPO_4P/gVFACOD$ and 1.0 gPHA/gVFACOD, respectively. The $NO_3N$ requirement for the phosphorus uptake as an electron acceptor was about $0.5 gNO_3N/gPO_4P_{uptake}$ based on the proposed equation with PHA, biomass, production, and the concentration of phosphorus release/uptake. Bacterial-community analysis of the sludge, as determined by FISH and 16SrDNA characterization FISH, revealed that the beta-subclass proteobacteria were the most abundant group ($27.9\%$ of the proteobacteria-specific probe EUB338), and it was likely that representative of the beta-subclass played key roles in activated sludge. The next dominant group found was the gamma-protebacteria ($15.4\%$ of probe EUB338). 16S rDNA clone library analysis showed that the members of${\beta}$- and ${\gamma}$-proteobacteria were also the most abundant groups, and $21.5\%$ (PN2 and PN4) and $15.4\%$ (PN1 and PN5) of total clones were the genera of denitrifying bacteria and PAO, respectively. Prediction of the microbial community composition was made with phosphorus content (Pv, $\%$ P/VSS) in wasted sludge and profiles of COD, PHA, $PO_4P,\;and\;NO_3N$ in an anaerobic-anoxic SBR unit. Generally, the predicted microbial composition based upon metabolic function, i.e., as measured by stoichiometry, is fairly similar to that measure by the unculturable dependent method. In this study, a proposal was made on he microbial community composition that was more easily approached to analyze the reactor behavior.

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들깻잎(Perilla frutescens)으로부터 U937 세포에 apoptosis를 유도하는 광과민성 물질의 분리 및 구조동정 (Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937)

  • 하준영;김미경;이준영;최은비;홍창오;이병원;배창환;김근기
    • 생명과학회지
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    • 제25권1호
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    • pp.53-61
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    • 2015
  • 본 연구에서 우리는 들깻잎(Perilla frutescens)으로부터 U937 세포에 대해 apoptosis를 유도하는 광과민성 물질을 분리하기 위해 연구를 시작하였다. 들깻잎은 한국의 대중적인 쌈채소로 바타민, GABA, 미네랄 등이 풍부하다고 알려져 있다. 건조한 들깻잎을 메탄올로 추출하여 다양한 크로마토그래피법을 이용하여 광과민성 물질을 순수 분리하였다. 순수 분리한 광과민성 물질을 PL9443으로 명명하고, 구조동정을 위하여 1D-NMR, 2D-NMR, FAB-mass spectroscopy 분석을 실시하였다. 순수 분리한 PL9443 compound의 흡광도를 측정한 결과 410 nm에서 최대 흡수를 보였고, 330, 410, 680 nm에서 흡수 peak를 나타내었다. PL9443 compound는 분자량 620.7의 pheophorbide a ethyl ester로 동정하였다. 구조동정한 물질은 porphyrin환을 갖는 구조에서 Mg이온이 탈리된 pheophorbide 유도체화합물이었다. PL9443 compound의 처리에 따른 U937 세포의 형태학적 변화를 관찰한 결과, 광조건에서 apoptosis의 전형적인 현상인 apoptotic body와 소낭구조물이 형성되는 것을 확인할 수 있었다. 또한 PL9443 compound를 0.08 nM로 처리하였을 때 99%의 caspase-3/7 activity가 나타났고, 세포손상 역시 광의존적으로 나타내는 것을 확인하였다. PL9443 compound를 처리하고 광을 조사한 처리구와 광을 조사하지 않은 처리구의 세포로부터 DNA를 분리하여 전기영동을 실시한 결과 광을 조사한 곳에서 DNA ladder현상을 관찰할 수 있었다. 따라서 PL9443 compound는 광의존적 apoptosis 유도에 의한 세포손상을 일으키는 것을 확인하였다.

Isolation and Characterization of Bacillus sp. Producing Broad-Spectrum Antibiotics Against Human and Plant Pathogenic Fungi

  • Chen, Na;Jin, Min;Qu, Hong-Mei;Chen, Zhi-Qiang;Chen, Zhao-Li;Qiu, Zhi-Gang;Wang, Xin-Wei;Li, Jun-Wen
    • Journal of Microbiology and Biotechnology
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    • 제22권2호
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    • pp.256-263
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    • 2012
  • A strain of bacterium producing antifungal antibiotic was isolated and identification of the strain was attempted. We could identify the bacterium as being a Bacillus sp., based on morphological observation, physiological characteristics, and 16S rDNA sequence analysis, thus leading us to designate the strain as Bacillus sp. AH-E-1. The strain showed potent antibiotic activity against phytopathogenic and human pathogenic fungi by inducing mycelial distortion and swelling and inhibiting spore germination. The antibiotic metabolite produced by the strain demonstrated excellent thermal and pH (2-11) stability, but was labile to autoclaving. From these results, we could find a broader antifungal activity of Bacillus genus. Isolation and characterization of the active agent produced by the strain are under progress.

Molecular Cloning of Maltooligosyltrehalose Trehalohydrolase Gene from Nostoc flagelliforme and Trehalose-Related Response to Stresses

  • Wu, Shuangxiu;He, Liang;Shen, Rongrong;Zhang, Xiu;Wang, Quanxi
    • Journal of Microbiology and Biotechnology
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    • 제21권8호
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    • pp.830-837
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    • 2011
  • A genomic DNA fragment encoding a putative maltooligosyltrehalose trehalohydrolase (NfMTH) for trehalose biosynthesis was cloned by the degenerate primer- PCR from cyanobacterium Nostoc flagelliforme. The ORF of NfMTH is 1,848 bp in length and encodes 615 amino acid residues, constituting a 70 kDa protein. The deduced amino acid sequence of NfMTH contains 4 regions highly conserved for MTHs. By expression of NfMTH in E. coli, the function of this protein was demonstrated, where the recombinant protein catalyzed the hydrolysis of maltooligosyl trehalose to trehalose. The expressions of MTH and maltooligosyltrehalose synthase in the filaments of N. flagelliforme were upregulated significantly under dehydration stress, NaCl stress, and high temperature-drought stress. The accumulations of both trehalose and sucrose in the filaments of N. flagelliforme were also improved significantly under the above stresses. Furthermore, trehalose accumulated in smaller quantities than sucrose did when under NaCl stress, but accumulated in higher quantities than sucrose did when under temperature-drought stress, indicating that both trehalose and sucrose were involved in N. flagelliforme adapted to stresses and different strategies conducted in response to various stress conditions.

Phylogenetic Characteristics of Echinococcus granulosus Sensu Lato in Uzbekistan

  • Kim, Hye-Jin;Yong, Tai-Soon;Shin, Myeong Heon;Lee, Kyu-Jae;Park, Gab-Man;Suvonkulov, Uktamjon;Kovalenko, Dmitriy;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • 제58권2호
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    • pp.205-210
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    • 2020
  • Echinococcosis occurs mainly in areas with heavy livestock farming, such as Central Asia, America, and Australia. Echinococcus granulosus sensu lato (s.l.) infection causes echinococcosis in intermediate hosts, such as sheep, cattle, goats, camels, and horses. Numerous cases of echinococcosis occur in Uzbekistan as stock farming is a primary industry. Epidemiological and genetic studies of E. granulosus s.l. are very important for mitigating its impact on public health and the economy; however, there are no such studies on E. granulosus s.l. in Uzbekistan. In the present study, to determine which genotypes exist and are transmitted, we isolated Echinococcus sp. from definitive hosts (one isolate each from jackal and dog) and intermediate hosts (52 isolates from humans and 6 isolates from sheep) in Uzbekistan and analyzed the isolates by sequencing 2 mitochondrial DNA components (cox1 and nad1). The results showed that all of isolates except one belonged to the E. granulosus sensu stricto (s.s.) G1 and G3 genotypes. Phylogenetic analysis based on cox1 sequences showed that 42 isolates from humans, 6 isolates from sheep, and one isolate from jackal were the G1 genotype, whereas the remaining 8 isolates from human and the one isolate from dog were the G3 genotype. These results suggest that the G1 and G3 genotypes of E. granulosus s.s. are predominant in Uzbekistan, and both wild animals and domestic animals are important for maintaining their life cycle. Only one isolate from human sample was confirmed to be E. eqiinus (G4 genotype), which is known to be for the first time.

Virulence, Resistance Genes, and Transformation Amongst Environmental Isolates of Escherichia coli and Acinetobacter spp.

  • Doughari, Hamuel James;Ndakidemi, Patrick Alois;Human, Izanne Susan;Benade, Spinney
    • Journal of Microbiology and Biotechnology
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    • 제22권1호
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    • pp.25-33
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    • 2012
  • The association of verotoxic E. coli and Acinetobacter spp. with various antibiotic-resistant, diarrhogenic, and nosocomial infections has been a cause for concern worldwide. E. coli and A. haemolyticus isolated on a number of selective media were screened for virulence factors, antibiotic resistance, and transformation of resistance genes. Out of 69 E. coli isolates obtained, 25 (35.23%), 14 (20.30%), and 28 (40.58%) were positive for Vtx1&2, Vtx1, and Vtx2, respectively, 49 (71.015%) for extendedspectrum beta-lactamases (ESBLs), 34 (49.28%) for serum resistance, 57 (82.61%) for cell surface hydrophobicity, 48 (69.57%) for gelatinase production, and 37 (53.62%) for hemolysin production. For the 14 A. haemolyticus isolates, only 2 (14.29%) in each case from all the samples investigated were positive for Vtx1, Vtx2 and Vtx1&2 respectively, 8 (57.14%) for ESBLs, 7 (50.00%) for serum resistance, 11 (78.57%) for cell surface hydrophobicity, 4 (28.57%) for gelatinase production, and 8 (57.14%) for hemolysin production. Although transformation occurred among the E. coli and Acinetobacter isolates (transformation frequency: $13.3{\times}10^{-7}-53.4^{-7}$), there was poor curing of the plasmid genes, a confirmation of the presence of stable antibiotic-resistant genes (DNA concentration between 42.7 and 123.8 ${\mu}g$) and intragenetic transfer of multidrug-resistant genes among the isolates. The isolates were potentially virulent and contained potentially transferable antibiotic resistance genes. Detection of virulence factors, antibiotic resistance genes, and transformation among these isolates is a very significant outcome that will influence approaches to proactive preventive and control measures and future investigations. However, continued surveillance for drug resistance among these bacteria and further investigation of the mechanism of action of their virulence factors are a necessity.

Enterococcus faecium LKE12 Cell-Free Extract Accelerates Host Plant Growth via Gibberellin and Indole-3-Acetic Acid Secretion

  • Lee, Ko-Eun;Radhakrishnan, Ramalingam;Kang, Sang-Mo;You, Young-Hyun;Joo, Gil-Jae;Lee, In-Jung;Ko, Jae-Hwan;Kim, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • 제25권9호
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    • pp.1467-1475
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    • 2015
  • The use of microbial extracts containing plant hormones is a promising technique to improve crop growth. Little is known about the effect of bacterial cell-free extracts on plant growth promotion. This study, based on phytohormonal analyses, aimed at exploring the potential mechanisms by which Enterococcus faecium LKE12 enhances plant growth in oriental melon. A bacterial strain, LKE12, was isolated from soil, and further identified as E. faecium by 16S rDNA sequencing and phylogenetic analysis. The plant growth-promoting ability of an LKE12 bacterial culture was tested in a gibberellin (GA)-deficient rice dwarf mutant (waito-C) and a normal GA biosynthesis rice cultivar (Hwayongbyeo). E. faecium LKE12 significantly improved the length and biomass of rice shoots in both normal and dwarf cultivars through the secretion of an array of gibberellins (GA1, GA3, GA7, GA8, GA9, GA12, GA19, GA20, GA24, and GA53), as well as indole-3-acetic acid (IAA). To the best of our knowledge, this is the first study indicating that E. faecium can produce GAs. Increases in shoot and root lengths, plant fresh weight, and chlorophyll content promoted by E. faecium LKE12 and its cell-free extract inoculated in oriental melon plants revealed a favorable interaction of E. faecium LKE12 with plants. Higher plant growth rates and nutrient contents of magnesium, calcium, sodium, iron, manganese, silicon, zinc, and nitrogen were found in cell-free extract-treated plants than in control plants. The results of the current study suggest that E. faecium LKE12 promotes plant growth by producing GAs and IAA; interestingly, the exogenous application of its cell-free culture extract can be a potential strategy to accelerate plant growth.