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      A Selected Core Microbiome Drives the Early Stages of Three Popular Italian Cheese Manufactures

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          Abstract

          Mozzarella (M), Grana Padano (GP) and Parmigiano Reggiano (PR) are three of the most important traditional Italian cheeses. In the three cheese manufactures the initial fermentation is carried out by adding natural whey cultures (NWCs) according to a back-slopping procedure. In this study, NWCs and the corresponding curds from M, GP and PR manufactures were analyzed by culture-independent pyrosequencing of the amplified V1–V3 regions of the 16S rRNA gene, in order to provide insights into the microbiota involved in the curd acidification. Moreover, culture-independent high-throughput sequencing of lacS gene amplicons was carried out to evaluate the biodiversity occurring within the S. thermophilus species. Beta diversity analysis showed a species-based differentiation between GP-PR and M manufactures indicating differences between the preparations. Nevertheless, all the samples shared a naturally-selected core microbiome, that is involved in the curd acidification. Type-level variability within S. thermophilus species was also found and twenty-eight lacS gene sequence types were identified. Although lacS gene did not prove variable enough within S. thermophilus species to be used for quantitative biotype monitoring, the possibility of using non rRNA targets for quantitative biotype identification in food was highlighted.

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          Rapid denoising of pyrosequencing amplicon data: exploiting the rank-abundance distribution

          We developed a fast method for denoising pyrosequencing for community 16S rRNA analysis. We observe a 2–4 fold reduction in the number of observed OTUs (operational taxonomic units) comparing denoised with non-denoised data. ~50,000 sequences can be denoised on a laptop within an hour, two orders of magnitude faster than published techniques. We demonstrate the effects of denoising on alpha and beta diversity of large 16S rRNA datasets.
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            Streptococcus suis: an emerging zoonotic pathogen.

            Streptococcus suis is a major porcine pathogen worldwide, and can be transmitted to human beings by close contact with sick or carrier pigs. S suis causes meningitis, septicaemia, endocarditis, arthritis, and septic shock in both pigs and human beings, and mortality is high. Human infection with S suis occurs mainly among certain risk groups that have frequent exposure to pigs or pork. Outbreaks of human S suis infection are uncommon, although several outbreaks have occurred in China in recent years. In July, 2005, the largest outbreak of human S suis infection occurred in Sichuan province, China, where 204 people were infected and 38 of them died. There have been 409 cases of human S suis infection worldwide, most of which have occurred in China, Thailand, and the Netherlands, and these infections have led to 73 deaths. This review provides background information on the biology and molecular characteristics of this Gram-positive bacterium, and describes the clinical signs, pathology, epidemiology, diagnosis, and treatment of human infection with S suis.
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              High-throughput sequencing and metagenomics: moving forward in the culture-independent analysis of food microbial ecology.

              Following recent trends in environmental microbiology, food microbiology has benefited from the advances in molecular biology and adopted novel strategies to detect, identify, and monitor microbes in food. An in-depth study of the microbial diversity in food can now be achieved by using high-throughput sequencing (HTS) approaches after direct nucleic acid extraction from the sample to be studied. In this review, the workflow of applying culture-independent HTS to food matrices is described. The current scenario and future perspectives of HTS uses to study food microbiota are presented, and the decision-making process leading to the best choice of working conditions to fulfill the specific needs of food research is described.
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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, USA )
                1932-6203
                2014
                24 February 2014
                : 9
                : 2
                : e89680
                Affiliations
                [1 ]Division of Microbiology, Department of Agricultural Sciences, University of Naples Federico II, Portici, Italy
                [2 ]Department of Food Science, University of Parma, Parma, Italy
                [3 ]Multidisciplinary Interdepartmental Dairy Center - MILC, University of Parma, Parma, Italy
                Teagasc Food Research Centre, Ireland
                Author notes

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

                Conceived and designed the experiments: DE. Performed the experiments: FDF ALS GS MG. Analyzed the data: FDF DE. Contributed reagents/materials/analysis tools: DE. Wrote the paper: FDF DE.

                Article
                PONE-D-13-50961
                10.1371/journal.pone.0089680
                3933672
                24586960
                afe443dd-5639-4530-8821-3cccc7315309
                Copyright @ 2014

                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
                : 4 December 2013
                : 22 January 2014
                Page count
                Pages: 8
                Funding
                This work was supported by a grant from the “Ministero dell’Istruzione, dell’Università e della Ricerca” within the program PON01_02863 Research and Competitiveness 2007–2013. Project title: “Encapsulation of active ingredients for improving the quality and food safety”. Francesca De Filippis was supported by a grant from Campania Region within the program “POR CAMPANIA FSE 2007/2013” - project CARINA (Safety sustainability and competitiveness of the agro-food production in Campania). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Agriculture
                Agricultural production
                Biology
                Biotechnology
                Applied microbiology
                Computational biology
                Genomics
                Genome sequencing
                Ecology
                Community ecology
                Community structure
                Biodiversity
                Microbial ecology
                Microbiology
                Applied microbiology
                Industrial microbiology
                Microbial ecology
                Veterinary science
                Veterinary microbiology

                Uncategorized
                Uncategorized

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