• Title/Summary/Keyword: 16S rRNA analysis

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Identification of bacteria isolated from rockworm viscera and application of isolated bacteria to shrimp aquaculture wastewater treatment

  • Ja Young Cho;Kyoung Sook Cho;Chang Hoon Kim;Joong Kyun Kim
    • Korean Journal of Environmental Biology
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    • v.41 no.2
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    • pp.167-178
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    • 2023
  • Large amounts of waste and wastewater from aquaculture have negatively impacted ecosystems. Among them, shrimp aquaculture wastewater contains large amounts of nitrogen contaminants derived from feed residues in an aerobic environment. This study isolated candidate strains from adult rockworms to treat shrimp aquaculture wastewater (SAW) in an aerobic environment. Among 87 strains isolated, 25 grew well at the same temperature as the shrimp aquaculture with excellent polymer degradation ability (>0.5 cm clear zone). Six isolates (strains AL1, AL4, AL5, AL6, LA10, and PR15) were finally selected after combining strains with excellent polymer degradation ability without antagonism. 16S rRNA sequencing analysis revealed that strains AL1, AL4, AL5, AL6, LA10, and PR15 were closely related to Bacillus paramycoides, Bacillus pumilus, Stenotrophomonas rhizophila, Bacillus paranthracis, Bacillus paranthracis, and Micrococcus luteus, respectively. When these six isolates were applied to SAW, they reached a maximum cell viability of 2.06×105 CFU mL-1. Their chemical oxygen demand (CODCr) and total nitrogen(TN) removal rates for 12h were 51.0% and 44.6%, respectively, when the CODCr/TN ratio was approximately 10.0. Considering these removal rates achieved in this study under batch conditions, these six isolates could be used for aerobic denitrification. Consequently, these six isolates from rockworms are good candidates that can be applied to the field of aquaculture wastewater treatment.

Rhizospheric-Derived Nocardiopsis alba BH35 as an Effective Biocontrol Agent Actinobacterium with Antifungal and Plant Growth-Promoting Effects: In Vitro Studies

  • Mohamed H. El-Sayed;Abd El-Nasser A. Kobisi;Islam A. Elsehemy;Mohamed A. El-Sakhawy
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.607-620
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    • 2023
  • The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 ㎍/ml) and ammonia (20 ㎍/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.

Composition and functional diversity of bacterial communities during swine carcass decomposition

  • Michelle Miguel;Seon-Ho Kim;Sang-Suk Lee;Yong-Il Cho
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1453-1464
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    • 2023
  • Objective: This study investigated the changes in bacterial communities within decomposing swine microcosms, comparing soil with or without intact microbial communities, and under aerobic and anaerobic conditions. Methods: The experimental microcosms consisted of four conditions: UA, unsterilized soil-aerobic condition; SA, sterilized soil-aerobic condition; UAn, unsterilized soil-anaerobic condition; and San, sterilized soil-anaerobic condition. The microcosms were prepared by mixing 112.5 g of soil and 37.5 g of ground carcass, which were then placed in sterile containers. The carcass-soil mixture was sampled at day 0, 5, 10, 30, and 60 of decomposition, and the bacterial communities that formed during carcass decomposition were assessed using Illumina MiSeq sequencing of the 16S rRNA gene. Results: A total of 1,687 amplicon sequence variants representing 22 phyla and 805 genera were identified in the microcosms. The Chao1 and Shannon diversity indices varied in between microcosms at each period (p<0.05). Metagenomic analysis showed variation in the taxa composition across the burial microcosms during decomposition, with Firmicutes being the dominant phylum, followed by Proteobacteria. At the genus level, Bacillus and Clostridium were the main genera within Firmicutes. Functional prediction revealed that the most abundant Kyoto encyclopedia of genes and genomes metabolic functions were carbohydrate and amino acid metabolisms. Conclusion: This study demonstrated a higher bacteria diversity in UA and UAn microcosms than in SA and SAn microcosms. In addition, the taxonomic composition of the microbial community also exhibited changes, highlighting the impact of soil sterilization and oxygen on carcass decomposition. Furthermore, this study provided insights into the microbial communities associated with decomposing swine carcasses in microcosm.

Characterization of Cholesterol Lowering Lactic Acid Bacteria Isolated from Palm Wine and Maize Beer and Assessment of Their Use in the Production of Probiotic Papaya Juice

  • Bertrand Tatsinkou Fossi;Dickson Ebwelle Ekabe;Liliane Laure Toukam Tatsinkou;Rene Bilingwe Ayiseh;Frederic Tavea;Pierre Michel Jazet
    • Microbiology and Biotechnology Letters
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    • v.51 no.2
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    • pp.191-202
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    • 2023
  • Elevated serum cholesterol is a main risk factor for heart disorders. Most probiotic products administered to lower cholesterol are dairy products which are not suitable for lactose-intolerant individuals. In this study, we assessed the cholesterol-lowering efficacy of LAB isolated from traditionally fermented drinks in diet-induced rats and determine their efficacy in the production of non-dairy, probiotic formulations using papaya juice. LAB were isolated from palm wine and corn beer on MRS agar using a pour-plate technique. Identification was carried out using 16S rRNA gene sequencing. A hypercholesterolemia model in which diet-induced Wistar albino rats were assigned into four groups was established. Oral gavage was carried out for 30 days. On the 31st day, the rats were dissected and the serum lipid profile was analyzed using biochemical kits. A 106 cfu/ml of a 24-h-old culture of selected lactobacilli was used to inoculate papaya juice and incubated at 37℃. Microbial and chemical changes were assessed during papaya fermentation and after four weeks of cold storage. Two selected isolates (Pw1 and Cb4) had in vitro cholesterol reduction of > 80%. These two isolates lowered lipid profile (triglyceride, total cholesterol, LDL-c) significantly, and increased HDL-c levels (p < 0.5) in the rat sera. Phylogenetic analysis showed that Pw1 was 98.86% similar to Limosilactobacillus fermentum, while Cb4 was 99.54% similar to Enteroccocus faecium. Both strains fermented papaya juice with cell viability reaching 8.92 × 108 cfu/ml and 25.3 × 108 cfu/ml respectively, and were still viable after 4 weeks of cold storage.

Biological Control of Fusarium oxysporum, the Causal Agent of Fusarium Basal Rot in Onion by Bacillus spp.

  • Jong-Hwan Shin;Ha-Kyoung Lee;Seong-Chan Lee;You-Kyoung Han
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.600-613
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    • 2023
  • Fusarium oxysporum is the main pathogen causing Fusarium basal rot in onion (Allium cepa L.), which incurs significant yield losses before and after harvest. Among management strategies, biological control is an environmentally safe and sustainable alternative to chemical control. In this study, we isolated and screened bacteria for antifungal activity against the basal rot pathogen F. oxysporum. Isolates 23-045, 23-046, 23-052, 23-055, and 23-056 significantly inhibited F. oxysporum mycelial growth and conidial germination. Isolates 23-045, 23-046, 23-052, and 23-056 suppressed the development of Fusarium basal rot in both onion seedlings and bulbs in pot and spray inoculation assays. Isolate 23-055 was effective in onion seedlings but exhibited weak inhibitory effect on onion bulbs. Based on analyses of the 16S rRNA and rpoB gene sequences together with morphological analysis, isolates 23-045, 23-046, 23-052, and 23-055 were identified as Bacillus thuringiensis, and isolate 23-056 as Bacillus toyonensis. All five bacterial isolates exhibited cellulolytic, proteolytic, and phosphate-solubilizing activity, which may contribute to their antagonistic activity against onion basal rot disease. Taken together B. thuringiensis 23-045, 23-046, 23-052, and 23-055 and B. toyonensis 23-056 have potential for the biological control of Fusarium basal rot in onion.

Association between LEPR Genotype and Gut Microbiome in Healthy Non-Obese Korean Adults

  • Yoon Jung Cha;In Ae Chang;Eun-Heui Jin;Ji Hye Song;Jang Hee Hong;Jin-Gyu Jung;Jung Sunwoo
    • Biomolecules & Therapeutics
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    • v.32 no.1
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    • pp.146-153
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    • 2024
  • The LEPR (leptin receptor) genotype is associated with obesity. Gut microbiome composition differs between obese and non-obese adults. However, the impact of LEPR genotype on gut microbiome composition in humans has not yet been studied. In this study, the association between LEPR single nucleotide polymorphism (rs1173100, rs1137101, and rs790419) and the gut microbiome composition in 65 non-obese Korean adults was investigated. Leptin, triglyceride, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol levels were also measured in all participants. Mean ± SD (standard deviation) of age, body mass index, and leptin hormone levels of participants was 35.2 ± 8.1 years, 21.4 ± 1.8 kg/m2, and 7989.1 ± 6687.4 pg/mL, respectively. Gut microbiome analysis was performed at the phylum level by 16S rRNA sequencing. Among the 11 phyla detected, only one showed significantly different relative abundances between LEPR genotypes. The relative abundance of Candidatus Saccharibacteria was higher in the G/A genotype group than in the G/G genotype group for the rs1137101 single nucleotide polymorphism (p=0.0322). Participant characteristics, including body mass index, leptin levels, and other lipid levels, were similar between the rs1137101 G/G and G/A genotypes. In addition, the relative abundances of Fusobacteria and Tenericutes showed significant positive relationship with plasma leptin concentrations (p=0.0036 and p=0.0000, respectively). In conclusion, LEPR genotype and gut microbiome may be associated even in normal-weight Korean adults. However, further studies with a greater number of obese adults are needed to confirm whether LEPR genotype is related to gut microbiome composition.

Profiles of Bacillus spp. Isolated from the Rhizosphere of Suaeda glauca and Their Potential to Promote Plant Growth and Suppress Fungal Phytopathogens

  • Lu, Ping;Jiang, Ke;Hao, Ya-Qiao;Chu, Wan-Ying;Xu, Yu-Dong;Yang, Jia-Yao;Chen, Jia-Le;Zeng, Guo-Hong;Gu, Zhou-Hang;Zhao, Hong-Xin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1231-1240
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    • 2021
  • Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

Isolation and Characterization of Bacillus Species Having Antifungal Activity Against Pathogens of Ginseng Damping Off (인삼모잘록병원균에 항균활성을 갖는 Bacillus 균의 분리 및 특성조사)

  • Park, Kyeong Hun;Park, Hong Woo;Lee, Seong Woo;Lee, Seung Ho;Myung, Kyung Sun;Lee, Sang Yeob;Song, Jaekyeong;Kim, Young Tak;Park, Kyoung Soo;Kim, Young Ock
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.380-387
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    • 2016
  • This study was performed to select potentially available biological control agent from soil bacteria for prevention of ginseng damping off. More than five hundred strains were isolated from ginseng rhizosphere soil. By testing antifungal activity, we have selected three soil bacteria strains and their ability to produce antibiotics and lytic enzymes such as cellulase, protease and pectate lyase was examined. Also, the presence of genes for biosynthesis of lipopeptide such as fengycin, bacillomycin D, surfactin, iturin A, and zwittermicin A was investigated in selected strains. All three strains produced cellulase, protease, and xylanase. Moreover, these strains had gene for biosynthesis of bacillomycin D, surfactin, and iturin A. ES1 and ES3 strains were identified Bacillus methylotrophucus and ES2 was confirmed Bacillus amyloliquefaciens using phylogenetic analysis on the basis of 16S rRNA gene sequences. In field test, control value of ES1, ES2 and ES3 treatment was 32.4%, 46.8% and 36.7%, respectively. This results indicate that antagonistic microbes with high ability of antifungal and lytic enzyme activity can be used as a useful biological control agent to control ginseng damping off.

Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.23 no.3
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    • pp.415-425
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    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Enhanced biological effect of fermented soy-powder milk with Lactobacillus brevis increasing in γ-aminobutyric acid and isoflavone aglycone contents (가바와 비당체 이소플라본이 증가된 Lactobacillus brevis 발효 콩-분말 두유의 생리활성 증진 효과)

  • Hwang, Chung Eun;Kim, Su Cheol;Lee, Jin Hwan;Hong, Su Young;Cho, Kye Man
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.245-255
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    • 2018
  • The research was aimed to analyze the functional constituents (GABA and isoflavone), radical (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl) scavenging activities and enzyme (${\alpha}-glucosidase$ and lipase) inhibitory effects of soypowder milk (SPM) and fermented soy-powder milk (FSPM) with varied Lactobacillus brevis. Ten ${\gamma}-aminobutyric$ acid (GABA) producing lactic acid bacteria that showed 96-99% similarity with L. brevis, according to 16S rRNA gene sequence analysis, were isolated from fermented kimchi. The conversion rates of GABA were obtained 66.96-93.51, 63.76-84.58, and 57.05-69.75% in monosodium glutamate, gluten and soy protein, respectively. The levels of pH and glutamic acid of FSPM were found lower than those of SPM, but the acidity and GABA contents were higher. The GABA conversion rate of FSPM with BMK484 strain was attained the highest 69.97%. The contents of isoflavone glycoside ($1290.93{\mu}g/g$) was higher in SPM, but the content of isoflavone aglycone ($287.27-501.9{\mu}g/g$) was higher in FSPM. The levels of isoflavone aglycone such as daidzein, glycitein and genistein, were found as the highest 240.2, 61.24 and $200.45{\mu}g/g$, respectively, when FSPM was made with BMK484 strain. The DPPH, ABTS and hydroxyl radical scavenging and ${\alpha}-glucosidase$ and pancreatic lipase inhibitory activities of FSPM made with BMK484 strain were the relatively higher 60.31, 88.10, 61.25, 52.71, and 39.37%, respectively. Therefore, the L. brevis can be used as a material capable of simultaneously enhanced GABA and isoflavone aglycone in FSPM.