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      Food authentication from shotgun sequencing reads with an application on high protein powders

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          Abstract

          Here we propose that using shotgun sequencing to examine food leads to accurate authentication of ingredients and detection of contaminants. To demonstrate this, we developed a bioinformatic pipeline, FASER (Food Authentication from SEquencing Reads), designed to resolve the relative composition of mixtures of eukaryotic species using RNA or DNA sequencing. Our comprehensive database includes >6000 plants and animals that may be present in food. FASER accurately identified eukaryotic species with 0.4% median absolute difference between observed and expected proportions on sequence data from various sources including sausage meat, plants, and fish. FASER was applied to 31 high protein powder raw factory ingredient total RNA samples. The samples mostly contained the expected source ingredient, chicken, while three samples unexpectedly contained pork and beef. Our results demonstrate that DNA/RNA sequencing of food ingredients, combined with a robust analysis, can be used to find contaminants and authenticate food ingredients in a single assay.

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          Most cited references21

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          Large-scale contamination of microbial isolate genomes by Illumina PhiX control

          With the rapid growth and development of sequencing technologies, genomes have become the new go-to for exploring solutions to some of the world’s biggest challenges such as searching for alternative energy sources and exploration of genomic dark matter. However, progress in sequencing has been accompanied by its share of errors that can occur during template or library preparation, sequencing, imaging or data analysis. In this study we screened over 18,000 publicly available microbial isolate genome sequences in the Integrated Microbial Genomes database and identified more than 1000 genomes that are contaminated with PhiX, a control frequently used during Illumina sequencing runs. Approximately 10% of these genomes have been published in literature and 129 contaminated genomes were sequenced under the Human Microbiome Project. Raw sequence reads are prone to contamination from various sources and are usually eliminated during downstream quality control steps. Detection of PhiX contaminated genomes indicates a lapse in either the application or effectiveness of proper quality control measures. The presence of PhiX contamination in several publicly available isolate genomes can result in additional errors when such data are used in comparative genomics analyses. Such contamination of public databases have far-reaching consequences in the form of erroneous data interpretation and analyses, and necessitates better measures to proofread raw sequences before releasing them to the broader scientific community.
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            A DNA Mini-Barcoding System for Authentication of Processed Fish Products

            Species substitution is a form of seafood fraud for the purpose of economic gain. DNA barcoding utilizes species-specific DNA sequence information for specimen identification. Previous work has established the usability of short DNA sequences—mini-barcodes—for identification of specimens harboring degraded DNA. This study aims at establishing a DNA mini-barcoding system for all fish species commonly used in processed fish products in North America. Six mini-barcode primer pairs targeting short (127–314 bp) fragments of the cytochrome c oxidase I (CO1) DNA barcode region were developed by examining over 8,000 DNA barcodes from species in the U.S. Food and Drug Administration (FDA) Seafood List. The mini-barcode primer pairs were then tested against 44 processed fish products representing a range of species and product types. Of the 44 products, 41 (93.2%) could be identified at the species or genus level. The greatest mini-barcoding success rate found with an individual primer pair was 88.6% compared to 20.5% success rate achieved by the full-length DNA barcode primers. Overall, this study presents a mini-barcoding system that can be used to identify a wide range of fish species in commercial products and may be utilized in high throughput DNA sequencing for authentication of heavily processed fish products.
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              Authentication of Herbal Supplements Using Next-Generation Sequencing

              Background DNA-based testing has been gaining acceptance as a tool for authentication of a wide range of food products; however, its applicability for testing of herbal supplements remains contentious. Methods We utilized Sanger and Next-Generation Sequencing (NGS) for taxonomic authentication of fifteen herbal supplements representing three different producers from five medicinal plants: Echinacea purpurea, Valeriana officinalis, Ginkgo biloba, Hypericum perforatum and Trigonella foenum-graecum. Experimental design included three modifications of DNA extraction, two lysate dilutions, Internal Amplification Control, and multiple negative controls to exclude background contamination. Ginkgo supplements were also analyzed using HPLC-MS for the presence of active medicinal components. Results All supplements yielded DNA from multiple species, rendering Sanger sequencing results for rbcL and ITS2 regions either uninterpretable or non-reproducible between the experimental replicates. Overall, DNA from the manufacturer-listed medicinal plants was successfully detected in seven out of eight dry herb form supplements; however, low or poor DNA recovery due to degradation was observed in most plant extracts (none detected by Sanger; three out of seven–by NGS). NGS also revealed a diverse community of fungi, known to be associated with live plant material and/or the fermentation process used in the production of plant extracts. HPLC-MS testing demonstrated that Ginkgo supplements with degraded DNA contained ten key medicinal components. Conclusion Quality control of herbal supplements should utilize a synergetic approach targeting both DNA and bioactive components, especially for standardized extracts with degraded DNA. The NGS workflow developed in this study enables reliable detection of plant and fungal DNA and can be utilized by manufacturers for quality assurance of raw plant materials, contamination control during the production process, and the final product. Interpretation of results should involve an interdisciplinary approach taking into account the processes involved in production of herbal supplements, as well as biocomplexity of plant-plant and plant-fungal biological interactions.
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                Author and article information

                Contributors
                +1 914 945 1394 , nshaimin@us.ibm.com
                klbeck@us.ibm.com
                Journal
                NPJ Sci Food
                NPJ Sci Food
                NPJ Science of Food
                Nature Publishing Group UK (London )
                2396-8370
                19 November 2019
                19 November 2019
                2019
                : 3
                : 24
                Affiliations
                [1 ]Consortium for Sequencing the Food Supply Chain, San Jose, CA USA
                [2 ]GRID grid.481554.9, IBM T.J. Watson Research Center, ; Yorktown Heights, NY USA
                [3 ]GRID grid.481551.c, IBM Almaden Research Center, ; San Jose, CA USA
                [4 ]ISNI 0000 0004 1936 9684, GRID grid.27860.3b, University of California Davis, School of Veterinary Medicine, ; Davis, CA USA
                [5 ]Mars Global Food Safety Center, Beijing, China
                [6 ]Wisdom Health, A Division of Mars Petcare, Vancouver, WA USA
                [7 ]GRID grid.467419.9, Mars Incorporated, ; McLean, VA USA
                [8 ]ISNI 0000 0001 2187 1663, GRID grid.418312.d, Bio-Rad Laboratories, ; Hercules, CA USA
                [9 ]ISNI 0000 0004 0623 3323, GRID grid.481801.4, Bio-Rad, Food Science Division, ; Marnes-La-Coquette, France
                Author information
                http://orcid.org/0000-0002-8663-1019
                http://orcid.org/0000-0003-2786-2447
                http://orcid.org/0000-0003-0171-5166
                http://orcid.org/0000-0002-7471-1978
                http://orcid.org/0000-0001-7835-881X
                http://orcid.org/0000-0001-8696-0405
                http://orcid.org/0000-0002-7049-7134
                http://orcid.org/0000-0002-7872-5074
                http://orcid.org/0000-0002-4603-0235
                Article
                56
                10.1038/s41538-019-0056-6
                6863864
                31754632
                1c2fedb8-6feb-47c5-9f0c-2fe9a1dd78aa
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 21 March 2019
                : 23 August 2019
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                © The Author(s) 2019

                metagenomics,food microbiology
                metagenomics, food microbiology

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