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Victoria M Pearson, S Brian Caudle, Darin R Rokyta
Understanding the structure and dynamics of microbial communities, especially those of economic concern, is of paramount importance to maintaining healthy and efficient microbial communities at agricultural sites and large industrial cultures, including bioprocessors. Wastewater treatment plants are large bioprocessors which receive water from multiple sources, becoming reservoirs for the collection of many viral families that infect a broad range of hosts. To examine this complex collection of viruses, full-length genomes of circular ssDNA viruses were isolated from a wastewater treatment facility using a combination of sucrose-gradient size selection and rolling-circle amplification and sequenced on an Illumina MiSeq...
2016: PeerJ
Ilona F Persoon, Mark J Buijs, Ahmet R Özok, Wim Crielaard, Bastiaan P Krom, Egija Zaura, Bernd W Brandt
OBJECTIVES: Bacterial infection of the root canal system causes apical periodontitis. Less is known about the role of fungi in these infections. This study aimed to assess the fungal prevalence, abundance, and diversity of root canal infections, as well as the relation between fungi and bacteria present in different parts of the root canal. MATERIALS AND METHODS: Twenty-six teeth with primary apical periodontitis were extracted, split in apical and coronal root segments, and cryo-pulverized...
October 22, 2016: Clinical Oral Investigations
James Heng Chiak Sim, Cynthia Truong, Samuel S Minot, Nick Greenfield, Indre Budvytiene, Akshar Lohith, Victoria Anikst, Nader Pourmand, Niaz Banaei
Understanding the contribution of relapse and reinfection to recurrent Clostridium difficile infection (CDI) has implications for therapy and infection prevention, respectively. We used whole genome sequencing to determine the relation of C. difficile strains isolated from patients with recurrent CDI at an academic medical center in the United States. Thirty-five toxigenic C. difficile isolates from 16 patients with 19 recurrent CDI episodes with median time of 53.5days (range, 13-362) between episodes were whole genome sequenced on the Illumina MiSeq platform...
October 5, 2016: Diagnostic Microbiology and Infectious Disease
Maksym Misyura, Tong Zhang, Mahadeo A Sukhai, Mariam Thomas, Swati Garg, Suzanne Kamel-Reid, Tracy L Stockley
Use of next-generation sequencing to detect somatic variants in DNA extracted from formalin-fixed, paraffin-embedded tumor tissues poses a challenge for clinical molecular diagnostic laboratories because of variable DNA quality and quantity, and the potential to detect low allele frequency somatic variants difficult to verify by non-next-generation sequencing methods. We evaluated somatic variant detection performance of the MiSeq and Ion Proton benchtop sequencers using two commercially available panels, the TruSeq Amplicon Cancer Panel and the AmpliSeq Cancer Hotspot Panel Version 2...
November 2016: Journal of Molecular Diagnostics: JMD
Fuqin Zou, Dong Zeng, Bin Wen, Hao Sun, Yi Zhou, Mingyue Yang, Zhirong Peng, Shuai Xu, Hesong Wang, Xiangchao Fu, Dan Du, Yan Zeng, Hui Zhu, Kangcheng Pan, Bo Jing, Ping Wang, Xueqin Ni
Antibiotics have been widely used for the prevention and the treatment of diseases to humans and animals, and they have fed additives for agricultural animals to promote growth. However, there is a growing concern over the practice due to its side effects on intestinal microbial communities which plays a vital role in animals' health. To investigate the effect of antibiotics on the bacterial population of the caecum in rex rabbits, 80 rex rabbits were randomly divided into four groups: control group (B, basal diet), chlortetracycline group (C, 50 mg/kg), colistin sulfate group (S, 20 mg/kg) and zinc bacitracin group (Z, 40 mg/kg)...
December 2016: AMB Express
Liyan Song, Shu Yang, Hongjie Liu, Jing Xu
Little is known regarding how bacterial communities assemble at landfill, as well as how the environment shapes the composition of bacterial community. In this study, up to 42 refuse samples from a large-scale landfill in China were physicochemically and phylogenetically investigated. 16S ribosomal RNA (rRNA) gene-based Illumina MiSeq sequencing (nine samples) revealed that representatives of Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Bacteroidetes were dominant in the refuse samples, which was similar to a previous study on landfill leachate by using 454 pyrosequencing...
October 21, 2016: Applied Microbiology and Biotechnology
Ya Qin, Jie Hou, Ming Deng, Quansheng Liu, Chongwei Wu, Yingjie Ji, Xugang He
The abundance and diversity of bacteria in two types of ponds were investigated by quantitative PCR and Illumina MiSeq sequencing. The results revealed that the abundance of bacterial 16S rRNA genes in D ponds (with grass carp fed sudan grass) was significantly lower than that in E ponds (with grass carp fed commercial feed). The microbial communities were dominated by Proteobacteria, Cyanobacteria, Bacteroidetes, and Actinobacteria in both E and D ponds, while the abundance of some genera was significantly different between the two types of ponds...
October 19, 2016: Scientific Reports
Valentina Kovaleva, Anna-Lena Geissler, Lisa Lutz, Ralph Fritsch, Frank Makowiec, Sebastian Wiesemann, Ulrich T Hopt, Bernward Passlick, Martin Werner, Silke Lassmann
BACKGROUND: Targeted next generation sequencing (tNGS) has become part of molecular pathology diagnostics for determining RAS mutation status in colorectal cancer (CRC) patients as predictive tool for decision on EGFR-targeted therapy. Here, we investigated mutation profiles of case-matched tissue specimens throughout the disease course of CRC, to further specify RAS-status dynamics and to identify de novo mutations associated with distant metastases. METHODS: Case-matched formalin-fixed and paraffin-embedded (FFPE) resection specimens (n = 70; primary tumours, synchronous and/or metachronous liver and/or lung metastases) of 14 CRC cases were subjected to microdissection of normal colonic epithelial, primary and metastatic tumour cells, their DNA extraction and an adapted library protocol for limited DNA using the 48 gene TruSeq Amplicon Cancer Panel(TM), MiSeq sequencing and data analyses (Illumina)...
October 18, 2016: Molecular Cancer
Habteab Habtom, Sandrine Demanèche, Lorna Dawson, Chen Azulay, Ofra Matan, Patrick Robe, Ron Gafny, Pascal Simonet, Edouard Jurkevitch, Zohar Pasternak
The ubiquity and transferability of soil makes it a resource for the forensic investigator, as it can provide a link between agents and scenes. However, the information contained in soils, such as chemical compounds, physical particles or biological entities, is seldom used in forensic investigations; due mainly to the associated costs, lack of available expertise, and the lack of soil databases. The microbial DNA in soil is relatively easy to access and analyse, having thus the potential to provide a powerful means for discriminating soil samples or linking them to a common origin...
October 6, 2016: Forensic Science International. Genetics
Xihui Xu, Zhou Zhang, Shunli Hu, Zhepu Ruan, Jiandong Jiang, Chen Chen, Zhenguo Shen
Soil provides a critical environment for microbial community development. However, microorganisms may be sensitive to substances such as heavy metals (HMs), which are common soil contaminants. This study investigated bacterial communities using 16S ribosomal RNA (rRNA) gene fragment sequencing in geographic regions with and without HM pollution to elucidate the effects of soil properties and HMs on bacterial communities. No obvious changes in the richness or diversity of bacterial communities were observed between samples from mining and control areas...
October 15, 2016: Environmental Science and Pollution Research International
Asma Asemaninejad, R Greg Thorn, Zoë Lindo
Peatlands play an important role in global climate change through sequestration of atmospheric CO2. Climate-driven changes in the structure of fungal communities in boreal peatlands that favor saprotrophic fungi can substantially impact carbon dynamics and nutrient cycling in these crucial ecosystems. In a mesocosm study using a full factorial design, 100 intact peat monoliths, complete with living Sphagnum and above-ground vascular vegetation, were subjected to three climate change variables (increased temperature, reduced water table, and elevated CO2 concentrations)...
October 15, 2016: Microbial Ecology
Ping Lin, Bingyu Ding, Chunyan Feng, Shuwei Yin, Ting Zhang, Xin Qi, Huiying Lv, Xiaokui Guo, Ke Dong, Yongzhang Zhu, Qingtian Li
BACKGROUND: The diagnosis of major depression disorder (MDD) and other mental disorders were depended on some subjective survey scales. There are confirmed relationship between the gut flora and the mental states of MDD patients. METHODS: The V3-V4 region of the 16S rRNA gene was extracted from the fecal microbial communities in MDD patients, PCR amplified and sequenced on the Illumina Miseq platform. RESULTS: More phylum Firmicutes, less Bacteroidetes, and more genus Prevotella, Klebsiella, Streptococcus and Clostridium XI were found in MDD patients...
October 1, 2016: Journal of Affective Disorders
Teresa Graziano, Angela Amoruso, Stefania Nicola, Francesca Deidda, Serena Allesina, Marco Pane, Pietro Piffanelli, Francesco Strozzi, Luca Mogna, Mario Del Piano
GOALS: The aim of the study was to unequivocally demonstrate the nontransmissibility of the genes mediating the resistance of the strain Bifidobacterium longum W11 (LMG P-21586) to rifaximin. BACKGROUND: Most antibiotic treatments can induce unfavorable side effects such as antibiotic-associated diarrhea, which is largely attributable to the disruption of the intestinal microbiota. The parallel intake of probiotic bacteria might reduce these events, even if with generally very poor results...
November 2016: Journal of Clinical Gastroenterology
Dorsaf Kerfahi, Ryunosuke Tateno, Koichi Takahashi, HyunJun Cho, Hyoki Kim, Jonathan M Adams
There is considerable interest in understanding the processes of microbial development in volcanic ash. We tested the predictions that there would be (1) a distinctive bacterial community associated with soil development on volcanic ash, including groups previously implicated in weathering studies; (2) a slower increase in bacterial abundance and soil C and N accumulation in cooler climates; and (3) a distinct communities developing on the same substrate in different climates. We set up an experiment, taking freshly fallen, sterilized volcanic ash from Sakurajima volcano, Japan...
October 12, 2016: Microbial Ecology
Jing Han, Peng Wang, Shaohua Ge
The object of this study is to characterize the bacterial community of subgingival plaque of two subjects with generalized aggressive periodontitis (GAgP) pre- and post-treatment. We picked two patients with GAgP and used high-throughput 16S rDNA sequencing. V4 hypervariable region was picked for PCR amplification of subgingival samples. Then, the PCR products were sequenced through Illumina MiSeq platform. One month after therapy, both the clinical features and periodontal parameters improved obviously. Moreover, the composition and structure of subgingival bacterial community changed after initial periodontal therapy...
October 8, 2016: Oncotarget
Yad Ghavi-Helm, Bingqing Zhao, Eileen E M Furlong
Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) is an invaluable technique to assess transcription factor binding and histone modifications in a genome-wide manner, an essential step towards understanding the mechanisms that govern embryonic development. Here, we provide a detailed protocol for all steps involved in generating a ChIP-seq library, starting from embryo collection, fixation, chromatin preparation, immunoprecipitation, and finally library preparation. The protocol is optimized for Drosophila embryos, but can be easily adapted for any model organism...
2016: Methods in Molecular Biology
Jan Hubert, Martina Bicianova, Ondrej Ledvinka, Martin Kamler, Philip J Lester, Marta Nesvorna, Jan Kopecky, Tomas Erban
The honey bee, Apis mellifera, is a globally important species that suffers from a variety of pathogens and parasites. These parasites and pathogens may have sublethal effects on their bee hosts via an array of mechanisms, including through a change in symbiotic bacterial taxa. Our aim was to assess the influence of four globally widespread parasites and pathogens on the honey bee bacteriome. We examined the effects of the ectoparasitic mite Varroa destructor, the fungal pathogens Nosema apis and Nosema ceranae, and the trypanosome Lotmaria passim...
October 11, 2016: Microbial Ecology
Yue Yuan, Jinjin Liu, Bin Ma, Ye Liu, Bo Wang, Yongzhen Peng
This study presents a novel strategy to improve the removal efficiency of nitrogen and phosphorus from municipal wastewater by feeding sequencing batch reactor (SBR) with sludge alkaline fermentation products as carbon sources. The performances of two SBRs treating municipal wastewater (one was fed with sludge fermentation products; F-SBR, and the other without sludge fermentation products; B-SBR) were compared. The removal efficiencies of total nitrogen (TN) and phosphorus (PO4(3-)-P) were found to be 82.9% and 96...
October 8, 2016: Bioresource Technology
Liliang Guo, Zhenghong Sui, Yuan Liu
Miseq sequencing and data analysis for the actin gene and v9 region of 18S rDNA of 7 simulated samples consisting of different mixture of dinoflagellates and diatoms were carried out. Not all the species were detectable in all the 18S v9 samples, and sequence percent in all the v9 samples were not consistent with the corresponding cell percent which may suggest that 18S rDNA copy number in different cells of these species differed greatly which result in the large deviation of the amplification. And 18S rDNA amplification of the microalgae was prone to be contaminated by fungus...
October 10, 2016: Scientific Reports
Yaohui Bai, Yangyang Chang, Jinsong Liang, Chen Chen, Jiuhui Qu
High concentrations of iron (Fe(II)) and manganese (Mn(II)) often occur simultaneously in groundwater. Previously, we demonstrated that Fe(II) and Mn(II) could be oxidized to biogenic Fe-Mn oxides (BFMO) via aeration and microbial oxidation, and the formed BFMO could further oxidize and adsorb other pollutants (e.g., arsenic (As(III)) and antimony (Sb(III))). To apply this finding to groundwater remediation, we established four quartz-sand columns for treating groundwater containing Fe(II), Mn(II), As(III), and Sb(III)...
September 21, 2016: Water Research
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