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      Inferring microbial interactions in thermophilic and mesophilic anaerobic digestion of hog waste

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          Abstract

          Anaerobic digestion (AnD) is a microbiological process that converts organic waste materials into biogas. Because of its high methane content, biogas is a combustible energy source and serves as an important environmental technology commonly used in the management of animal waste generated on large animal farms. Much work has been done on hardware design and process engineering for the generation of biogas. However, little is known about the complexity of the microbiology in this process. In particular, how microbes interact in the digester and eventually breakdown and convert organic matter into biogas is still regarded as a “black box.” We used 16S rRNA sequencing as a tool to study the microbial community in laboratory hog waste digesters under tightly controlled conditions, and systematically unraveled the distinct interaction networks of two microbial communities from mesophilic (MAnD) and thermophilic anaerobic digestion (TAnD). Under thermophilic conditions, the well-known association between hydrogen-producing bacteria, e.g., Ruminococcaceae and Prevotellaceae, and hydrotrophic methanogens, Methanomicrobiaceae, was reverse engineered by their interactive topological niches. The inferred interaction network provides a sketch enabling the determination of microbial interactive relationships that conventional strategy of finding differential taxa was hard to achieve. This research is still in its infancy, but it can help to depict the dynamics of microbial ecosystems and to lay the groundwork for understanding how microorganisms cohabit in the anaerobic digester.

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          Most cited references51

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          FLASH: fast length adjustment of short reads to improve genome assemblies.

          Next-generation sequencing technologies generate very large numbers of short reads. Even with very deep genome coverage, short read lengths cause problems in de novo assemblies. The use of paired-end libraries with a fragment size shorter than twice the read length provides an opportunity to generate much longer reads by overlapping and merging read pairs before assembling a genome. We present FLASH, a fast computational tool to extend the length of short reads by overlapping paired-end reads from fragment libraries that are sufficiently short. We tested the correctness of the tool on one million simulated read pairs, and we then applied it as a pre-processor for genome assemblies of Illumina reads from the bacterium Staphylococcus aureus and human chromosome 14. FLASH correctly extended and merged reads >99% of the time on simulated reads with an error rate of <1%. With adequately set parameters, FLASH correctly merged reads over 90% of the time even when the reads contained up to 5% errors. When FLASH was used to extend reads prior to assembly, the resulting assemblies had substantially greater N50 lengths for both contigs and scaffolds. The FLASH system is implemented in C and is freely available as open-source code at http://www.cbcb.umd.edu/software/flash. t.magoc@gmail.com.
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            Group-specific primer and probe sets to detect methanogenic communities using quantitative real-time polymerase chain reaction.

            Real-time polymerase chain reaction (PCR) is a highly sensitive method that can be used for the detection and quantification of microbial populations without cultivating them in anaerobic processes and environmental samples. This work was conducted to design primer and probe sets for the detection of methanogens using a real-time PCR with the TaqMan system. Six group-specific methanogenic primer and probe sets were designed. These sets separately detect four orders (Methanococcales, Methanobacteriales, Methanomicrobiales, and Methanosarcinales) along with two families (Methanosarcinaceae and Methanosaetaceae) of the order Methanosarcinales. We also designed the universal primer and probe sets that specifically detect the 16S rDNA of prokaryotes and of the domain Bacteria and Archaea, and which are fully compatible with the TaqMan real-time PCR system. Target-group specificity of each primer and probe set was empirically verified by testing DNA isolated from 28 archaeal cultures and by analyzing potential false results. In general, each primer and probe set was very specific to the target group. The primer and probe sets designed in this study can be used to detect and quantify the order-level (family-level in the case of Methanosarcinales) methanogenic groups in anaerobic biological processes and various environments.
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              Diversity and succession of the intestinal bacterial community of the maturing broiler chicken.

              The diversity of bacterial floras in the ilea and ceca of chickens that were fed a vegetarian corn-soy broiler diet devoid of feed additives was examined by analysis of 1,230 partial 16S rRNA gene sequences. Nearly 70% of sequences from the ileum were related to those of Lactobacillus, with the majority of the rest being related to Clostridiaceae (11%), Streptococcus (6.5%), and Enterococcus (6.5%). In contrast, Clostridiaceae-related sequences (65%) were the most abundant group detected in the cecum, with the other most abundant sequences being related to Fusobacterium (14%), Lactobacillus (8%), and Bacteroides (5%). Statistical analysis comparing the compositions of the different 16S rRNA libraries revealed that population succession occurred during some sampling periods. The significant differences among cecal libraries at 3 and 7 days of age, at 14 to 28 days of age, and at 49 days of age indicated that successions occurred from a transient community to one of increasing complexity as the birds aged. Similarly, the ileum had a stable bacterial community structure for birds at 7 to 21 days of age and between 21 to 28 days of age, but there was a very unique community structure at 3 and 49 days of age. It was also revealed that the composition of the ileal and cecal libraries did not significantly differ when the birds were 3 days old, and in fact during the first 14 days of age, the cecal microflora was a subset of the ileal microflora. After this time, the ileum and cecum had significantly different library compositions, suggesting that each region developed its own unique bacterial community as the bird matured.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                21 July 2017
                2017
                : 12
                : 7
                : e0181395
                Affiliations
                [1 ] Biodiversity Research Center, Academia Sinica, Taipei, Taiwan
                [2 ] Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan
                [3 ] Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taipei, Taiwan
                Kyungpook National University, REPUBLIC OF KOREA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: DW CYC.

                • Data curation: GTWS.

                • Formal analysis: GTWS.

                • Funding acquisition: DW CYC.

                • Investigation: DW GTWS CYW ACL CYC.

                • Methodology: DW GTWS CYW ACL CYC.

                • Project administration: DW CYC.

                • Resources: GTWS CYW ACL.

                • Supervision: DW.

                • Visualization: DW GTWS.

                • Writing – original draft: DW GTWS.

                • Writing – review & editing: DW GTWS.

                Author information
                http://orcid.org/0000-0003-3859-9162
                Article
                PONE-D-17-05759
                10.1371/journal.pone.0181395
                5521784
                28732056
                8af33815-766a-446b-8f2a-fa8c46f6c7ff
                © 2017 Shaw et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 18 February 2017
                : 2 July 2017
                Page count
                Figures: 5, Tables: 3, Pages: 22
                Funding
                Funded by: Sustainability Science Research Project, Academia Sinica.
                Award Recipient :
                This work was supported by Sustainability Science Research Project, Academia Sinica. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Ecology and Environmental Sciences
                Extremophiles
                Engineering and Technology
                Energy and Power
                Bioenergy
                Biofuels
                Biogas
                Engineering and Technology
                Energy and Power
                Fuels
                Biofuels
                Biogas
                Physical Sciences
                Materials Science
                Materials by Attribute
                Fuels
                Biofuels
                Biogas
                Biology and Life Sciences
                Organisms
                Archaeans
                Methanogens
                Physical Sciences
                Chemistry
                Chemical Compounds
                Methane
                Computer and Information Sciences
                Network Analysis
                Centrality
                Biology and Life Sciences
                Organisms
                Animals
                Vertebrates
                Amniotes
                Mammals
                Swine
                Biology and life sciences
                Biochemistry
                Nucleic acids
                RNA
                Non-coding RNA
                Ribosomal RNA
                Biology and life sciences
                Biochemistry
                Ribosomes
                Ribosomal RNA
                Biology and life sciences
                Cell biology
                Cellular structures and organelles
                Ribosomes
                Ribosomal RNA
                Biology and Life Sciences
                Ecology
                Community Ecology
                Trophic Interactions
                Ecology and Environmental Sciences
                Ecology
                Community Ecology
                Trophic Interactions
                Custom metadata
                All NGS sequence data analyzed during this study are available from the NCBI's Short Read Archive sequence database (BioProject accession number: PRJNA350355).

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                Uncategorized

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