scholarly journals Comprehensive Understanding of the Bacterial Populations and Metabolites Profile of Fermented Feed by 16S rRNA Gene Sequencing and Liquid Chromatography–Mass Spectrometry

Metabolites ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 239
Author(s):  
Wei Jin ◽  
Zheng Zhang ◽  
Kun Zhu ◽  
Yanfeng Xue ◽  
Fei Xie ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Antimicrobial Activities ◽  
Rrna Gene ◽  
Liquid Chromatography Mass Spectrometry ◽  
Bacterial Populations ◽  
Fermented Feed ◽  
Rrna Gene Sequencing

The comprehensive bacterial populations and metabolites profile in fermented feed is unclear, which may have significant effects on the stability of fermented feed quality and animal gut health. In this study, 16S rRNA gene sequencing and liquid chromatography–mass spectrometry were used to explore the bacterial populations and metabolites profile in the fermented feed incubated with probiotics (MF) or without probiotics (SF). The probiotics were a combination of Lactobacillus salivarius, Bacillus subtilis, and Saccharomyces cerevisiae. The pH and lactic acid levels were higher in MF than in SF (P < 0.05), while the total volatile fatty acid content was lower (P < 0.05). Interestingly, after fermentation, the most abundant bacterial genus in MF was Enterococcus, rather than the added probiotics Lactobacillus or Bacillus. Weissella and a few potential pathogens (Enterobacter, Escherichia-Shigella, and Pantoea) were dominant in SF (P < 0.05). Metabolomics analysis identified 32 different metabolites in the two types of fermented feed. These metabolites enriched in MF, such as maleic acid, phenylacetic acid, ethyl linoleate, dihomo-gamma-linolenic acid, and L-theanine had potential antimicrobial activities. Conclusively, the addition of probiotics enriched a few potentially beneficial microbes and small molecular compounds with antimicrobial activities, and inhibited the potential pathogens in fermented feed.

scholarly journals Comparison of the Vitek MS and Bruker Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Chryseobacterium Isolates from Clinical Specimens and Report of Uncommon Chryseobacterium Infections in Humans

2018 ◽  
Vol 56 (11) ◽  
Author(s):  
Jiun-Nong Lin ◽  
Shih-Hua Teng ◽  
Chung-Hsu Lai ◽  
Chih-Hui Yang ◽  
Yi-Han Huang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Flight Mass Spectrometry ◽  
Rrna Gene Sequencing ◽  
Vitek Ms

ABSTRACT Matrix-assisted laser desorption ionization–time of flight mass spectrometry is becoming more popular and is replacing traditional identification methods in the clinical microbiology laboratory. We aimed to compare the Vitek mass spectrometry (MS) and Bruker Biotyper systems for the identification of Chryseobacterium isolated from clinical specimens and to report uncommon Chryseobacterium infections in humans. The microbial database from a hospital was searched for records between 2005 and 2016 to identify cultures that yielded Chryseobacterium. Species identification by the Vitek MS and Bruker Biotyper systems was compared to identification by 16S rRNA gene sequencing. Over the study period, 140 Chryseobacterium isolates were included. Based on 16S rRNA gene sequencing, 78 isolates were C. indologenes, 39 were C. gleum, 12 were uncommon Chryseobacterium species (C. arthrosphaerae, C. culicis, C. cucumeris, C. bernardetii, C. artocarpi, and C. daecheongense), and the remaining 11 isolates were only identified at the genus level. The Vitek MS and Bruker Biotyper systems correctly identified 98.7% and 100% of C. indologenes isolates, respectively. While the Bruker Biotyper accurately identified 100% of C. gleum isolates, the Vitek MS system correctly identified only 2.6% of isolates from this species. None of the uncommon Chryseobacterium species were successfully identified by either of these two systems. The overall accuracies of Chryseobacterium identification at the species level by the Vitek MS and Bruker Biotyper systems were 60.5% and 90.7%, respectively. An upgrade and correction of the Vitek MS and Bruker Biotyper databases is recommended to correctly identify Chryseobacterium species.

scholarly journals Metaproteomic and 16S rRNA Gene Sequencing Analysis of the Infant Fecal Microbiome

2019 ◽  
Vol 20 (6) ◽  
pp. 1430 ◽  
Author(s):  
Laetitia Cortes ◽  
Harm Wopereis ◽  
Aude Tartiere ◽  
Julie Piquenot ◽  
Joost Gouw ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Sequencing Analysis ◽  
Microbial Composition ◽  
Fecal Microbiome ◽  
Rrna Gene Sequencing

A metaproteomic analysis was conducted on the fecal microbiome of eight infants to characterize global protein and pathway expression. Although mass spectrometry-based proteomics is now a routine tool, analysis of the microbiome presents specific technical challenges, including the complexity and dynamic range of member taxa, the need for well-annotated metagenomic databases, and high inter-protein sequence redundancy and similarity. In this study, an approach was developed for assessment of biological phenotype and metabolic status, as a functional complement to DNA sequence analysis. Fecal samples were prepared and analysed by tandem mass spectrometry and a homology-based meta-clustering strategy was used to combine peptides from multiple species into representative proteins. In total, 15,250 unique peptides were sequenced and assigned to 2154 metaclusters, which were then assigned to pathways and functional groups. Differences were noted in several pathways, consistent with the dominant genera observed in different subjects. Although this study was not powered to draw conclusions from the comparisons, the results obtained demonstrate the applicability of this approach and provide the methods needed for performing semi-quantitative comparisons of human fecal microbiome composition, physiology and metabolism, as well as a more detailed assessment of microbial composition in comparison to 16S rRNA gene sequencing.

scholarly journals Biotransformation of Patulin by Gluconobacter oxydans

2006 ◽  
Vol 73 (3) ◽  
pp. 785-792 ◽  
Author(s):  
A. Ricelli ◽  
F. Baruzzi ◽  
M. Solfrizzo ◽  
M. Morea ◽  
F. P. Fanizzi
Keyword(s):  
Mass Spectrometry ◽  
Gene Sequencing ◽  
Gluconobacter Oxydans ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Tandem Mass ◽  
Toxic Compound ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Rrna Gene Sequencing

ABSTRACT A bacterium isolated from patulin-contaminated apples was capable of degrading patulin to a less-toxic compound, ascladiol. The bacterium was identified as Gluconobacter oxydans by 16S rRNA gene sequencing, whereas ascladiol was identified by liquid chromatography-tandem mass spectrometry and proton and carbon nuclear magnetic resonance. Degradation of up to 96% of patulin was observed in apple juices containing up to 800 μg/ml of patulin and incubated with G. oxydans.

Screening and Molecular Characterization of Cellulase Producing Actinobacteria from Litchi Orchard

2019 ◽  
Vol 13 (1) ◽  
pp. 90-101
Author(s):  
Sanju Kumari ◽  
Utkarshini Sharma ◽  
Rohit Krishna ◽  
Kanak Sinha ◽  
Santosh Kumar
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Characterization ◽  
Biochemical Characterization ◽  
Evolutionary Relationship ◽  
Gene Sequencing ◽  
Cellulase Activity ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Rrna Gene Sequencing

Background: Cellulolysis is of considerable economic importance in laundry detergents, textile and pulp and paper industries and in fermentation of biomass into biofuels. Objective: The aim was to screen cellulase producing actinobacteria from the fruit orchard because of its requirement in several chemical reactions. Methods: Strains of actinobacteria were isolated on Sabouraud’s agar medium. Similarities in cultural and biochemical characterization by growing the strains on ISP medium and dissimilarities among them perpetuated to recognise nine groups of actinobacteria. Cellulase activity was measured by the diameter of clear zone around colonies on CMC agar and the amount of reducing sugar liberated from carboxymethyl cellulose in the supernatant of the CMC broth. Further, 16S rRNA gene sequencing and molecular characterization were placed before NCBI for obtaining recognition with accession numbers. Results: Prominent clear zones on spraying Congo Red were found around the cultures of strains of three groups SK703, SK706, SK708 on CMC agar plates. The enzyme assay for carboxymethylcellulase displayed extra cellulase activity in broth: 0.14, 0.82 and 0.66 &#181;mol mL-1 min-1, respectively at optimum conditions of 35°C, pH 7.3 and 96 h of incubation. However, the specific cellulase activities per 1 mg of protein did not differ that way. It was 1.55, 1.71 and 1.83 μmol mL-1 min-1. The growing mycelia possessed short compact chains of 10-20 conidia on aerial branches. These morphological and biochemical characteristics, followed by their verification by Bergey’s Manual, categorically allowed the strains to be placed under actinobacteria. Further, 16S rRNA gene sequencing, molecular characterization and their evolutionary relationship through phylogenetics also confirmed the putative cellulase producing isolates of SK706 and SK708 subgroups to be the strains of Streptomyces. These strains on getting NCBI recognition were christened as Streptomyces glaucescens strain SK91L (KF527284) and Streptomyces rochei strain SK78L (KF515951), respectively. Conclusion: Conclusive evidence on the basis of different parameters established the presence of cellulase producing actinobacteria in the litchi orchard which can convert cellulose into fermentable sugar.

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