• Title/Summary/Keyword: environmental DNA

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Applicability Evaluation of Male-Specific Coliphage-Based Detection Methods for Microbial Contamination Tracking

  • Kim, Gyungcheon;Park, Gwoncheol;Kang, Seohyun;Lee, Sanghee;Park, Jiyoung;Ha, Jina;Park, Kunbawui;Kang, Minseok;Cho, Min;Shin, Hakdong
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1709-1715
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    • 2021
  • Outbreaks of food poisoning due to the consumption of norovirus-contaminated shellfish continue to occur. Male-specific (F+) coliphage has been suggested as an indicator of viral species due to the association with animal and human wastes. Here, we compared two methods, the double agar overlay and the quantitative real-time PCR (RT-PCR)-based method, for evaluating the applicability of F+ coliphage-based detection technique in microbial contamination tracking of shellfish samples. The RT-PCR-based method showed 1.6-39 times higher coliphage PFU values from spiked shellfish samples, in relation to the double agar overlay method. These differences indicated that the RT-PCR-based technique can detect both intact viruses and non-particle-protected viral DNA/RNA, suggesting that the RT-PCR based method could be a more efficient tool for tracking microbial contamination in shellfish. However, the virome information on F+ coliphage-contaminated oyster samples revealed that the high specificity of the RT-PCR- based method has a limitation in microbial contamination tracking due to the genomic diversity of F+ coliphages. Further research on the development of appropriate primer sets for microbial contamination tracking is therefore necessary. This study provides preliminary insight that should be examined in the search for suitable microbial contamination tracking methods to control the sanitation of shellfish and related seawater.

Comparison of Cryoprotectants and Cryopreservation Protocols for Eleutherococcus senticosus via Somatic Embryogenesis

  • Ahn, Chang Ho;Shin, Jung Won;Lee, Ha Na;Yoon, Hyun Won;Seo, Jeong Min;Kim, Yeoung Ryul;Baek, Saeng Geul;Nam, Jae Ik;Choi, Yong Eui
    • Journal of Forest and Environmental Science
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    • v.38 no.3
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    • pp.152-158
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    • 2022
  • A cryopreservation is an essential tool for preservation of germplasm. In this study, the possibility for cryopreservation of embryogenic cells of Siberian ginseng (Eleutherococcus senticosus) in liquid nitrogen (-196℃) was evaluated. The effects of glycerol and dimethyl sulfoxide (DMSO) at different concentrations (5%, 10% and 20%) as cryoprotectants on regrowth of cryopreserved E. senticosus embryogenic cells were tested. There was significant effect of cryoprotectants on regrowth of embryogenic cells (p=0.0019). The highest and lowest fresh mass gain were achieved when embryogenic cells were frozen with 10% DMSO and 5% glycerol (138.2±5.9 and 61.3±14.6, respectively). The effect of the cryoprotectants on the frequency embryo germination was tested. There was no significant difference between glycerol and DMSO (p=0.846). Three different concentrations of cryoprotectants did not significantly affect the frequency embryo germination (p=0.534). Finally, the genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cells was tested by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. RAPD and ISSR analysises showed that there was no genetic variation among regenerants.

Genetic factors in precocious puberty

  • Shim, Young Suk;Lee, Hae Sang;Hwang, Jin Soon
    • Clinical and Experimental Pediatrics
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    • v.65 no.4
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    • pp.172-181
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    • 2022
  • Pubertal onset is known to result from reactivation of the hypothalamic-pituitary-gonadal (HPG) axis, which is controlled by complex interactions of genetic and nongenetic factors. Most cases of precocious puberty (PP) are diagnosed as central PP (CPP), defined as premature activation of the HPG axis. The cause of CPP in most girls is not identifiable and, thus, referred to as idiopathic CPP (ICPP), whereas boys are more likely to have an organic lesion in the brain. ICPP has a genetic background, as supported by studies showing that maternal age at menarche is associated with pubertal timing in their offspring. A gain of expression in the kisspeptin gene (KISS1), gain-of-function mutation in the kisspeptin receptor gene (KISS1R), loss-of-function mutation in makorin ring finger protein 3 (MKRN3), and loss-of-function mutations in the delta-like homolog 1 gene (DLK1) have been associated with ICPP. Other genes, such as gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1), lin-28 homolog B (LIN28B), neuropeptide Y (NPYR), tachykinin 3 (TAC3), and tachykinin receptor 3 (TACR3), have been implicated in the progression of ICPP, although their relationships require elucidation. Environmental and socioeconomic factors may also be correlated with ICPP. In the progression of CPP, epigenetic factors such as DNA methylation, histone posttranslational modifications, and non-coding ribonucleic acids may mediate the relationship between genetic and environmental factors. CPP is correlated with short- and long-term adverse health outcomes, which forms the rationale for research focusing on understanding its genetic and nongenetic factors.

Development & Assessment of Alkyl Chain Modified Aptamers as Potential PET Radiotracers for Lymphoma Diagnosis

  • Ji Woong Lee;Un Chol Shin;Seok u Bae;Ji Yoon Kim;Hae joon Cho;Ji Ae Park;Kyo Chul Lee;Jung Young Kim;Suhng Wook Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.8 no.2
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    • pp.77-85
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    • 2022
  • The Td05 and Sgc8c, DNA-based aptamers, are well-known to target internalized surface markers (IGHM and PTK7) of Burkitt's lymphoma and acute lymphoblastic leukemia (ALL). Thus, Td05 and Sgc8c labeled with metallic radioisotope 64Cu can be evaluated as potential diagnostic PET imaging agents. In this study, we modified the carbon chain length of the last adenosine of aptamer (n = 3, 6, 12) to increase tumor cell uptake and select the best candidate among six types of aptamer analogues and one adenosine of aptamer. After labeling of 64Cu, [64Cu]Cu-DOTA-aptamer analogues were evaluated in vitro studies (serum stability, Log P values, cell uptake, biodistribution). Then, we evaluate in vivo PET imaging study for two candidates (64Cu-DOTA-C12-Sgc8c, 64Cu-DOTA-C6-Td05). PET images clearly visualize tumors at 24 h post-injection rather than at an early time point and the tumor-to-background ratio also increases at the delay time point. 64Cu-DOTA-C12-Sgc8c and 64Cu-DOTA-C6-Td05 could be used as potential radiotracers for lymphoma.

Survey of Antibiotic Resistant Bacteria in Ulleungdo, Korea (울릉도의 항생제 내성균 조사)

  • Jun Hyung Lee;Hye Won Hong;Dukki Han
    • Korean Journal of Environmental Agriculture
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    • v.41 no.4
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    • pp.344-354
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    • 2022
  • BACKGROUND: Although antibiotics have contributed to treatment of bacterial infection, the antibiotic abuse can lead to antibiotic resistant bacteria. Impact of human activities on distribution of antibiotic resistance has been intensively issued and occurrence of antibiotic resistant bacteria in contaminated environments would not be a surprise. Nonetheless, anthropogenic contamination with the dissemination of antibiotic resistance along uncontaminated environments has been less considered. The aim of this study is to investigate antibiotic resistant bacteria across Ulleungdo, known as antibiotic resistance free and anthropogenic pollution free environment in Rep. of Korea. METHODS AND RESULTS: Antibiotic resistant bacteria in coastal seawater of Ulleungdo were investigated in July 2021. Antibiotic susceptibility test using the disk diffusion method was applied with six drugs according to the Clinical and Laboratory Standards Institute (CLSI) guideline. Total 43 bacterial isolates were tested and 20 isolates among of them showed multidrug resistance. Particularly, the number and ratio of resistant bacteria were relatively high in a densely populated area of Ulleungdo. The bacterial communities were investigated using 16S rRNA gene metabarcoding approach in the coastal seawater and soils of Ulleungdo. In the bacterial communities, Firmicutes were selectively distributed only in seawater, suggesting the possibility of anthropogenic contamination in coastal seawater of Ulleungdo. CONCLUSION(S): We found antibiotic resistant bacteria in a populated area of Ulleungdo. The occurrence of antibiotic resistant bacteria in Ulleungdo seems to result from the recent anthropogenic impact. Consistent monitoring of antibiotic resistant bacteria in the uncontaminated environment needs to considered for future risk assessment of antibiotics.

Molecular identification of fruit bats, natural host of Nipah virus in Bangladesh, based on DNA barcode

  • Md. Maharub Hossain Fahim;Walid Hassan;Afia Afsin;Md. Mahfuzur Rahman;Md. Tanvir Rahman;Sang Jin Lim;Yeonsu Oh;Yung Chul Park;Hossain Md. Faruquee;Md. Mafizur Rahman
    • Journal of Ecology and Environment
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    • v.48 no.2
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    • pp.163-172
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    • 2024
  • Background: Fruit bats are natural carriers of Nipah virus (NiV). The primary objective of this study is to identify potential reservoir species in a selected geographic regions. It is necessary to determine an accurate species identification of the associated reservoir bat species distributed in a specific region. Results: In this study, we collected 20 different bat specimens from the NiV-prone area of the Kushtia district. Among these, 14 were tissue samples (BT-1-14) and six were fecal samples (BF-1-6). We used the mitochondrial gene cytochrome b, one of the most abundant and frequently used genetic markers, for polymerase chain reaction amplification and sequencing. Out of the 20 samples, 12 tissue samples and 2 fecal samples were successfully amplified and sequenced. However, two tissue samples and four fecal samples yielded chimeric sequences, rendering them unsuitable for annotation. The sequences of the successfully amplified samples were compared to those deposited in the National Center for Biotechnology Information database using basic local alignment search tool to identify the bat specimen collected. The study identified six different bat species using both morphological and genetic data, which may carriers of the NiV. Conclusions: Our results suggest that additional research should be conducted to gather more information on fruit bats from different localities across the country. The study contributes to the establishment of appropriate measures for NiV carrying disease control and management.

Peltaster fructicola: Undescribed Sooty Blotch and Flyspeck Species on Apple Fruit in Korea

  • Jun-Woo Choi;Seong-Keun Lim;Seo-Ryeong Lee;Chang-Gi Back;In-Kyu Kang;Seung-Yeol Lee;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.52 no.2
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    • pp.145-153
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    • 2024
  • While investigation of the fungal diseases on apples collected from Cheongsong-gun and Bonghwa-gun in Gyeongbuk province, Korea, between August and September 2023 isolated five fungal strains from fruits with sooty blotch and flyspeck (SBFS) disease. The strains were designated as KNUF-23-CS02, KNUF-23-CS-06, KNUF-23-CS12, KNUF-23-BH01, and KNUF-23-BH03. When grown on potato dextrose agar and 2% water agar, the cultural characteristics of the strains were similar to those previously reported characteristics of Peltaster fructicola Pf001. The strains produced monoblastic, hyaline conidiogenous cells; the conidia were hyaline, unicellular, cylindrical to ovoidal, and 3.5-7×1.7-3.9 and 4.0-6.6×1.8-3.2 μm in size on synthetic nutrient-poor agar or water agars, respectively. Secondary conidia production by microcyclic conidiation and budding was observed. The KNUF-23-BH03 strain was shown to cause SBFS symptoms similar to those observed on the apples in the pathogenicity test. Molecular phylogenetic analyses were conducted based on the isolated species sequences of the internal transcribed spacer region, nuclear large ribosomal DNA subunit, and mitochondrial small ribosomal RNA subunit gene. The five strains were clustered with Peltaster fructicola Pf001. Based on the cultural and morphological characteristics and phylogenetic analysis, the five strains were identified as Peltaster fructicola, which has not been previously reported in Korea.

Biodiesel production using lipase producing bacteria isolated from button mushroom bed (양송이 배지에서 유래한 Lipase 생산균을 이용한 바이오디젤 생산)

  • Kim, Heon-Hee;Kim, Chan-Kyum;Han, Chang-Hoon;Lee, Chan-Jung;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.56-62
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    • 2015
  • A lipase producing bacterium was isolated from button mushroom bed, which showing high clear zone on agar media containing Tributyrin as the substrate. The strain was identified as Burkholderia cepacia by analysis of 16S rDNA gene sequence. Crude lipase (CL) was partially purified from 70% ammonium sulfate precipitation using the culture filtrate of B. cepacia. Immobilized lipases were prepared by cross-linking method with CL from B. cepacia and Novozyme lipase (NL) onto silanized Silica-gel as support. Residual activitiy of the immobilized CL (ICL) and immobilized NL (INL) was maintained upto 61% and 72%, respectively. Biodiesel (Fatty acid methyl ester, FAME) was recovered by transesterification and methanolysis of Canola oil using NaOH, CL and ICL as the catalysts to compare the composition of fatty acids and the yield of FAME. Total FAME content was NaOH $781mg\;L^{-1}$, CL $681mg\;L^{-1}$ and ICL $596mg\;L^{-1}$, in which the highest levels of FAME was observed to 50% oleic acid (C18:1) and 22% stearic acid (C18:0). In addition, the unsaturated FAME (C18:1, C18:2) decreased, while saturated FAME (C16:0, C18:0) increased according to increasing the reaction times with both CL and ICL, supporting CL possess both transesterification and interesterification activity. When reusability of ICL and INL was estimated by using the continuous reaction of 4 cycles, the activity of ICL and INL was respectively maintained 66% and 79% until the fourth reaction.

Cellulose degrading basidiomycetes yeast isolated from the gut of grasshopper in Korea (한국의 메뚜기의 장에서 분리된 Cellulose를 분해하는 담자균 효모)

  • Kim, Ju-Young;Jang, Jun Hwee;Park, Ji-Hyun;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Limtong, Savitree;Subramani, Gayathri;Sung, Gi-Ho;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.362-368
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    • 2018
  • Grasshoppers play vital role in the digestion of photosynthetically fixed carbons. With the aid of intestinal microflora, the grasshopper can degrade leaves constituents such as cellulose and hemicellulose. The purpose of this study was to examine cellulolytic yeast isolates from the gut of grasshoppers collected in Gyeonggi Province, South Korea. Among the yeast isolates, ON2, ON17 (two strains), and ON6 (one strain) showed positive cellulolytic activity in the CMC-plate assay. The sequence analyses of D1/D2 domains of the large subunit rDNA gene and the internal transcribed spacer (ITS) regions revealed that the strains ON2 and ON17 were most closely related to Papiliotrema aspenensis CBS $13867^T$ (100%, sequence similarity in D1/D2 domains; 99.4% sequence similarity in ITS) and strain ON6 related to Saitozyma flava (100% in D1/D2 domains; 99.0% in ITS). All these three yeast strains are capable of degrading cellulose; therefore, the members of endosymbiotic yeasts may produce their own enzymes for carbohydrate degradation and convert mobilized sugar monomers to volatile fatty acids. Thus, the endosymbiotic yeast strains ON2, ON17 (represents the genus Papilioterma) and ON6 (Saitozyma) belonging to the family Tremellomycetes, are unreported strains in Korea.

Cloning and Characterization of Two Distinct CD3 Genes from Olive Flounder Paralichthys olivaceus

  • Kim, Mu-Chan;Park, Chan-Il
    • Fisheries and Aquatic Sciences
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    • v.8 no.2
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    • pp.56-64
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    • 2005
  • Two distinct CD3 homologue genes, $CD3\gamma/\delta\;and\;CD\varepsilon$, were isolated from a olive flounder leukocyte cDNA library and a BAC library. $CD3\gamma/\delta$ consisted of 961 bp encoding 178 amino acid residues, and $CD3\varepsilon$ consisted of 1006 bp encoding 164 amino acid residues. When compared with other known CD3 peptide sequences, the most conserved region of the two olive flounder CD3 chain peptides are the cytoplasmic domain and the least conserved are the extracellular domain. A phylogenetic analysis based on the deduced amino acid sequence grouped the two olive flounder CD3 sequences with $CD3\varepsilon$ and $CD3\gamma/\delta$, respectively. The olive flounder CD3 cluster (consisting of $CD3\varepsilon\;and\;CD3\gamma/\delta$) spans only 10.4 kb. The $CD3\varepsilon\;and\;CD3\gamma/\delta$ genes are oppositely transcribed only 3.8 kb apart. Both olive flounder CD3 genes have five exons. The two olive flounder CD3 genes were predominantly expressed in PBLs, kidney, spleen, and gills.