• Title/Summary/Keyword: 16S-23S rRNA

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Identification of microbiome with 16S rRNA gene pyrosequencing and antimicrobial effect of egg white in bovine mastitis (젖소 유방염에서 16S rRNA 파이로시퀀싱을 이용한 우유 내 마이크로바이옴의 동정과 난백의 항균효과)

  • Kim, Danil;Kim, Eun-Kyung;Seong, Won-Jin;Ro, Younghye;Ko, Dae-Sung;Kim, Nam-Hyung;Kim, Jae-Hong;Kwon, Hyuk-Joon
    • Korean Journal of Veterinary Research
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    • v.57 no.2
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    • pp.117-126
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    • 2017
  • Bovine mastitis is an important microbial disease in the dairy industry. We investigated the frequencies of bacterial pathogens in 62 farms and pathogen antibiotic resistance from mastitis samples (n = 748). We tested the antimicrobial activity of chicken and duck egg white and lysozyme purified from chicken egg white. Moreover, we compared the microbiomes of normal and mastitic raw milk obtained by 16S rRNA gene pyrosequencing and culture methods. The results showed that the frequencies of Gram-positive pathogens (Enterococcus faecalis 37% and Staphylococcus aureus 36%) were higher than that of a Gram-negative pathogen (Escherichia coli 15%). Resistance frequencies to ampicillin and norfloxacin were lowest in Staphylococcus aureus (21%), Enterococcus faecalis (23%), and Escherichia coli (33%), and the antimicrobial activity of chicken egg white was higher than those of lysozyme and duck egg white. Pyrosequencing results revealed clear differences between the microbiomes of mastitic and normal raw milk samples and revealed a slightly similar, but clearly different, composition of pathogens compared to that from the culture method. Thus, pyrosequencing may be useful for elucidating changes in microbiomes during mastitis progression and treatment. A chicken egg white and antibiotic combination may help with mastitis treatment; however, further studies are needed.

Linking growth performance and carcass traits with enterotypes in Muscovy ducks

  • Qian Fan;Yini Xu;Yingping Xiao;Caimei Yang;Wentao Lyu;Hua Yang
    • Animal Bioscience
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    • v.37 no.7
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    • pp.1213-1224
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    • 2024
  • Objective: Enterotypes (ETs) are the clustering of gut microbial community structures, which could serve as indicators of growth performance and carcass traits. However, ETs have been sparsely investigated in waterfowl. The objective of this study was to identify the ileal ETs and explore the correlation of the ETs with growth performance and carcass traits in Muscovy ducks. Methods: A total of 200 Muscovy ducks were randomly selected from a population of 5,000 ducks at 70-day old, weighed and slaughtered. The growth performance and carcass traits, including body weight, dressed weight and evidenced weight, dressed percentage, percentage of apparent yield, breast muscle weight, leg muscle weight, percentage of leg muscle and percentage of breast muscle, were determined. The contents of ileum were collected for the isolation of DNA and 16S rRNA gene sequencing. The ETs were identified based on the 16S rRNA gene sequencing data and the correlation of the ETs with growth performance and carcass traits was performed by Spearman correlation analysis. Results: Three ETs (ET1, ET2, and ET3) were observed in the ileal microbiota of Muscovy ducks with significant differences in number of features and α-diversity among these ETs (p<0.05). Streptococcus, Candida Arthritis, and Bacteroidetes were the presentative genus in ET1 to ET3, respectively. Correlation analysis revealed that Lactococcus and Bradyrhizobium were significantly correlated with percentage of eviscerated yield and leg muscle weight (p<0.05) while ETs were found to have a close association with percentage of eviscerated yield, leg muscle weight, and percentage of leg muscle in Muscovy ducks. However, the growth performance of ducks with different ETs did not show significant difference (p>0.05). Lactococcus were found to be significantly correlated with leg muscle weight, dressed weight, and percentage of eviscerated yield. Conclusion: Our findings revealed a substantial variation in carcass traits associated with ETs in Muscovy ducks. It is implied that ETs might have the potential to serve as a valuable biomarker for assessing duck carcass traits. It would provide novel insights into the interaction of gut microbiota with growth performance and carcass traits of ducks.

Physiological Characteristics and Tyrosinase Inhibitory Activity of Lactobacillus plantarum M23 Isolated from Raw Milk (원유에서 분리한 Lactobacillus plantarum M23의 Tyrosinase 활성 저해 및 생리적 특성조사)

  • Heo, In-Sook;Kim, Kee-Sung;Yang, Seung-Yong;Lee, Nam-Hyouck;Lim, Sang-Dong
    • Food Science of Animal Resources
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    • v.27 no.4
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    • pp.501-508
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    • 2007
  • In order to develop a new starter for fermented milk, Lactobacillus plantarum M23 was isolated from raw milk and investigated for physiological characteristics. It showed good tyrosinase inhibitory activity compared with commercial lactic acid bacteria. The optimum growth temperature of Lactobacillus plantarum M23 was $40^{\circ}C$ and cultures took 17 hr to reach pH 4.3. Lactobacillus plantarum M23 showed more sensitivity to Penicillin-G, Oxacillin, Novobiocin, Chloramphenicol in a comparison of 12 different antibiotics, and showed most resistance to Vancomycin. It showed higher leucine arylamidase and ${\beta}-galactosidase$ activities compared to 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hours. It showed high resistance to Escherichia coli, Salmonella typhimurium and Staphylococcus aureus with rates of 77.8%, 86.5% and 83.8%, respectively. Based on these and previous results, Lactobacillus plantarum M23 could be an excellent starter culture for fermented milk with high resistance to melanin.

Comparison of Biotyping, Serotyping and Molecular Typing of Yersinia enterocolitica Isolated from Spring water in Seoul (서울시내 약수에서 분리한 Yersinia enterocolitica의 생물형, 혈청형 및 분자학적 형별비교)

  • 이영기;최성민;오수경;신재영
    • Journal of environmental and Sanitary engineering
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    • v.14 no.4
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    • pp.99-109
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    • 1999
  • Enteropathogenic Yersina enterocolitica is an important cause of human and animal disease. Phenotypic and genotypic characteristics currently used to identify Yersinia enterocolitica are not necessarily sufficient to differentiate pathogenic from non-pathogenic strains or to analyze the epidemiology of yersiniae at a molecular level. To improve the characterization of Yersinia enterocolitica, A total of 65 isolates of Yersinia enterocolitica were examined with bioserotyping, antibiotic susceptibilities, PFGE, PCR-ribotyping. Genomic DNA pattern generated by PFGE are highly specific for different strains of an organism and have significant value in epidemiologic investigations. The PFGE analysis of Not I-digested chromosomal DNA of Y. enterocolitica were performed with a CHEF Mapper(Bio-Rad, USA). Not I generated 19 restriction endonuclease digestion profiles(REDP). PCR-ribotyping, performed with primers complementry to conserved regions of 16S and 23S rRNA gene, generated 13 ribotypes. PCR-ribotyping can be considered a good technich for subtyping strains of Y.enterocolitica.

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Isolation of Mycoplasma pneumoniae and Antimicrobial Susceptibilities of the Isolates(III) (Mycoplasma pneumoniae의 분리 및 항생제 감수성 검사(III))

  • Chang Myung-Woong;Kim Kwang-Hyuk;Park In-Dal;Song Gap-Young;Kim Sung-Won;Lee Eun-young;Kim Moon-Chan;Cho Myung-Hoon;Kim Kyu-Earn;Choi Choong-Eon;Park Seon Yeong;Jo Hyeon Jang
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.479-485
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    • 2005
  • The 994 throat swabs obtained from 688 adults and 306 children patients with respiratory diseases were examined for Mycoplasma pneumoniae infection by culture method. Antimicrobial susceptibilities of the resulting 123 M. pneumoniae isolates were evaluated by testing minimum inhibitory concentrations (MICs) of erythromycin, minocycline, tetracycline, josamycin, sparfloxacin, ofloxacin, and ciprofloxacin by a broth micro-dilution method. The erythromycin resistant strains of M. pneumoniae was determined above $1.0{\mu}g/ml$ of MIC for erythromycin. The erythromycin resistant strains of M. pneumoniae was confirmed resistant gene mutation of the portions of genes 23S rRNA (domain II and V), and ribosomal protein 14 and L22 by PCR amplified and their nucleotide sequenses were compared to those of the susceptible strain M129. The isolation rate of M. pneumoniae was $12.9\%$ (89/688) for the adults and $11.1\%$ (34/306) for the children. The $MICs_{90}$ of the M. pneumoniae isolates were $0.12{\mu}g/ml$ for minocycline, $0.25{\mu}g/ml$ for sparfloxacin, $0.5{\mu}g/ml$ for ciprofloxacin, ofloxacin, and tetracycline, respectively, and $2.0{\mu}g/ml$ for josamycin and erythromycin, respectively. The isolation rate of erythromycin resistant M. pneumoniae from patients was $49.4\%\;(44/89)$ for the adults, $47.1\%\;(16/34)$ for children, and $48.8\%\;(60/123)$ for the total. No mutation could be detected in the ribosomal protein L22 region, but all strains were mutated in the ribosomal protein L4 as two point mutation M144V. Two point mutations in domain V of 23S rRNA were selected in the presense of erythromycin resistant M. pneumoniae isolates, such as one strain was G2057C mutant, two strains were A2059C mutants, three strains were C2611G mutants, four strains were A2058C mutants, five strains were A2058T mutants, twenty strains were A2059G mutants, and twenty-five strains were A2058G mutants, respectively. These results show that erythromycin was not the most active compound against M. pneumoniae infection in Korea and clinical studies of macrolides in human patients are demanded.

A report on 24 unrecorded bacterial species of Korea isolated in 2016, belonging to the orders Rhizobiales and Sphingomonadales in the class Alphaproteobacteria

  • Joung, Yochan;Cha, Chang-Jun;Im, Wan-Taek;Jeon, Che Ok;Joh, Kiseong;Kim, Seung-Bum;Kim, Wonyong;Lee, Soon Dong;Cho, Jang-Cheon
    • Journal of Species Research
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    • v.7 no.1
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    • pp.13-23
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    • 2018
  • In 2016, as a part of the research program 'Survey of Korean Indigenous Species', diverse environmental samples were collected from various sources of freshwater, seawater, soil, wetland, reclaimed land, sand, pine forest, plant root, ginseng field, solar saltern, and caves. Thousands of bacterial strains were isolated from the diverse samples and identified based on 16S rRNA gene sequence analyses. The present study, as a phylogenetic subset of the primary research program, reports 24 unrecorded bacterial species in Korea that belong to the orders Rhizobiales and Sphingomonadales in the class Alphaproteobacteria. Based on the high 16S rRNA gene sequence similarities (>98.8%) and formation of a robust phylogenetic clade with the closest type species, it was determined that each strain belonged to each independent and predefined bacterial species. There is no official report that these 24 bacterial species have been described in Korea; therefore, 10 species of nine genera in the order Rhizobiales and 14 species of seven genera in the order Sphingomonadales are described for unreported alphaproteobacterial species in Korea. Gram reaction, colony and cell morphology, biochemical properties, and isolation sources are also provided in the species description section.

A report of 35 unrecorded bacterial species isolated from sediment in Korea

  • Han, Ji-Hye;Baek, Kiwoon;Hwang, Seoni;Nam, Yoon Jong;Lee, Mi-Hwa
    • Journal of Species Research
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    • v.9 no.4
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    • pp.362-374
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    • 2020
  • A total of 35 bacterial strains were isolated from various sediment samples. From 16S rRNA gene sequence similarities higher than 98.7% and the formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to independent and predefined bacterial species. No previous official reports have described these 35 species in Korea. The unrecorded species were assigned to 6 phyla, 10 classes, 18 orders, 23 families, and 31 genera. At the genus level, the unrecorded species were affiliated with Terriglobus of the phylum Acidobacteria, as well as with Mycobacterium, Rhodococcus, Kineococcus, Phycicoccus, Agromyces, Cryobacterium, Microbacterium, and Arthrobacter; Catellatospora of the class Actinomycetia; Lacibacter of the class Chitinophagia; Algoriphagus and Flectobacillus of the class Cytophagia; Flavobacterium and Maribacter of the class Flavobacteriia; Bacillus, Cohnella, Fontibacillus, Paenibacillus, Lysynibacillus, and Paenisporosarcina of the class Bacilli; Bradyrhizobium, Gemmobacter, Loktanella, and Altererythrobacter of the class Alphaproteobacteria; Acidovorax of the class Betaproteobacteria; Aliiglaciecola, Cellvibrio, Arenimonas, and Lysobacter of class Gammaproteobacteria; and Roseimicrobium of the class Verrucomicrobia. The selected strains were subjected to further taxonomic characterization, including Gram reaction, cellular and colonial morphology, and biochemical properties. This paper provides detailed descriptions of the 35 previously unrecorded bacterial species.

Characterization of Canthaxanthin Isomers Isolated from a New Soil Dietzia sp. and Their Antioxidant Activities

  • Venugopalan, Vijayalatha;Tripathi, Subhash K.;Nahar, Pradip;Saradhi, P. Pardha;Das, Rakha H.;Gautam, Hemant K.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.237-245
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    • 2013
  • Canthaxanthin (cx) is a potent antioxidant that is chemically synthesized at the industrial scale and has imperative applications in the cosmetic and feed industries. An orange pigmented mesophilic bacterium, designated as K44, was isolated from soil samples of Kargil, India. Biochemical tests, 16S rRNA gene sequencing, and FAME analysis of the bacterium indicated it to belong in the genus Dietzia and is distinct from human isolates. The strain showed 98% 16S rRNA gene sequence homology with Dietzia maris DSM 43102. High-performance liquid chromatography profile of the pigments isolated from K44 showed two major peaks absorbing at 465.3 and 475 nm. The liquid chromatography-mass spectrometry (LC-MS) analysis of both these peaks revealed their m/z to be 564. The molecular weights, LC-MS/MS fragmentation patterns, and ${\lambda}_{max}$ of these fractions corresponded to all-trans- (475 nm) and 9-cis-(465.3 nm) cx isomers. The antioxidant activities of cis- and trans-cx isomers isolated from this bacterium were found to differ, where the cis-isomer showed higher free radical, superoxide radical, and reactive oxygen species scavenging activities than the alltrans- isomer, suggesting that 9-cis-cx is more effective as an antioxidant than the all-trans-cx.

Suppression of Fusarium Wilt Caused by Fusarium oxysporum f. sp. lactucae and Growth Promotion on Lettuce Using Bacterial Isolates

  • Yadav, Dil Raj;Adhikari, Mahesh;Kim, Sang Woo;Kim, Hyun Seung;Lee, Youn Su
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1241-1255
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    • 2021
  • This study was carried out to explore a non-chemical strategy for enhancing productivity by employing some antagonistic rhizobacteria. One hundred eighteen bacterial isolates were obtained from the rhizospheric zone of various crop fields of Gangwon-do, Korea, and screened for antifungal activity against Fusarium wilt (Fusarium oxysporum f. sp. lactucae) in lettuce crop under in vitro and in vivo conditions. In broth-based dual culture assay, fourteen bacterial isolates showed significant inhibition of mycelial growth of F. oxysporium f. sp. lactucae. All of the antagonistic isolates were further characterized for the antagonistic traits under in vitro conditions. The isolates were identified on the basis of biochemical characteristics and confirmed at their species level by 16S rRNA gene sequencing analysis. Arthrobacter sulfonivorans, Bacillus siamensis, Bacillus amyloliquefaciens, Pseudomonas proteolytica, four Paenibacillus peoriae strains, and Bacillus subtilis were identified from the biochemical characterization and 16S rRNA gene sequencing analysis. The isolates EN21 and EN23 showed significant decrease in disease severity on lettuce compared to infected control and other bacterial treatments under greenhouse conditions. Two bacterial isolates, EN4 and EN21, were evaluated to assess their disease reduction and growth promotion in lettuce in field conditions. The consortium of EN4 and EN21 showed significant enhancement of growth on lettuce by suppressing disease caused by F. oxysporum f. sp. lactucae respectively. This study clearly indicates that the promising isolates, EN4 (P. proteolytica) and EN21 (Bacillus siamensis), can be commercialized and used as biofertilizer and/or biopesticide for sustainable crop production.

Effects of Long-Term Fertilizer Practices on Rhizosphere Soil Autotrophic CO2-Fixing Bacteria under Double Rice Ecosystem in Southern China

  • Tang, Haiming;Wen, Li;Shi, Lihong;Li, Chao;Cheng, Kaikai;Li, Weiyan;Xiao, Xiaoping
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1292-1298
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    • 2022
  • Soil autotrophic bacterial communities play a significant role in the soil carbon (C) cycle in paddy fields, but little is known about how rhizosphere soil microorganisms respond to different long-term (35 years) fertilization practices under double rice cropping ecosystems in southern China. Here, we investigated the variation characteristics of rhizosphere soil RubisCO gene cbbL in the double rice ecosystems of in southern China where such fertilization practices are used. For this experiment we set up the following fertilizer regime: without any fertilizer input as a control (CK), inorganic fertilizer (MF), straw returning (RF), and organic and inorganic fertilizer (OM). We found that abundances of cbbL, 16S rRNA genes and RubisCO activity in rhizosphere soil with OM, RF and MF treatments were significantly higher than that of CK treatment. The abundances of cbbL and 16S rRNA genes in rhizosphere soil with OM treatment were 5.46 and 3.64 times higher than that of CK treatment, respectively. Rhizosphere soil RubisCO activity with OM and RF treatments increased by 50.56 and 45.22%, compared to CK treatment. Shannon and Chao1 indices for rhizosphere soil cbbL libraries with RF and OM treatments increased by 44.28, 28.56, 29.60, and 23.13% compared to CK treatment. Rhizosphere soil cbbL sequences with MF, RF and OM treatments mainly belonged to Variovorax paradoxus, uncultured proteobacterium, Ralstonia pickettii, Thermononospora curvata, and Azoarcus sp.KH33C. Meanwhile, cbbL-carrying bacterial composition was obviously influenced by soil bulk density, rhizosphere soil dissolved organic C, soil organic C, and microbial biomass C contents. Fertilizer practices were the principal factor influencing rhizosphere soil cbbL-carrying bacterial communities. These results showed that rhizosphere soil autotrophic bacterial communities were significantly changed under conditions of different long-term fertilization practices Therefore, increasing rhizosphere soil autotrophic bacteria community with crop residue and organic manure practices was found to be beneficial for management of double rice ecosystems in southern China.