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      Evaluation of fecal samples as a valid source of DNA by comparing paired blood and fecal samples from American bison ( Bison bison)

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          Abstract

          Background

          The collection and analysis of fecal DNA is a common practice, especially when dealing with wildlife species that are difficult to track or capture. While fecal DNA is known to be lower quality than traditional sources of DNA, such as blood or other tissues, few investigations have verified fecal samples as a valid source of DNA by directly comparing the results to high quality DNA samples from the same individuals. Our goal was to compare DNA from fecal and blood samples from the same 50 American plains bison ( Bison bison) from Yellowstone National Park, analyze 35 short tandem repeat (STR) loci for genotyping efficiency, and compare heterozygosity estimates.

          Results

          We discovered that some of the fecal-derived genotypes obtained were significantly different from the blood-derived genotypes from the same bison. We also found that fecal-derived DNA samples often underestimated heterozygosity values, in some cases by over 20%.

          Conclusions

          These findings highlight a potential shortcoming inherent in previous wildlife studies that relied solely on a multi-tube approach, using exclusively low quality fecal DNA samples with no quality control to account for false alleles and allelic dropout. Herein, we present a rigorous marker selection protocol that is applicable for a wide range of species and report a set of 15 STR markers for use in future bison studies that yielded consistent results from both fecal and blood-derived DNA.

          Electronic supplementary material

          The online version of this article (10.1186/s12863-019-0722-3) contains supplementary material, which is available to authorized users.

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          Most cited references30

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          Noninvasive genetic sampling: look before you leap.

          Noninvasive sampling allows genetic studies of free-ranging animals without the need to capture or even observe them, and thus allows questions to be addressed that cannot be answered using conventional methods. Initially, this sampling strategy promised to exploit fully the existing DNA-based technology for studies in ethology, conservation biology and population genetics. However, recent work now indicates the need for a more cautious approach, which includes quantifying the genotyping error rate. Despite this, many of the difficulties of noninvasive sampling will probably be overcome with improved methodology.
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            Molecular analysis of predation: a review of best practice for DNA-based approaches.

            Molecular analysis of predation, through polymerase chain reaction amplification of prey remains within the faeces or digestive systems of predators, is a rapidly growing field, impeded by a lack of readily accessible advice on best practice. Here, we review the techniques used to date and provide guidelines accessible to those new to this field or from a different molecular biology background. Optimization begins with field collection, sample preservation, predator dissection and DNA extraction techniques, all designed to ensure good quality, uncontaminated DNA from semidigested samples. The advantages of nuclear vs. mitochondrial DNA as primer targets are reviewed, along with choice of genes and advice on primer design to maximize specificity and detection periods following ingestion of the prey by the predators. Primer and assay optimization are discussed, including cross-amplification tests and calibratory feeding experiments. Once primers have been made, the screening of field samples must guard against (through appropriate controls) cross contamination. Multiplex polymerase chain reactions provide a means of screening for many different species simultaneously. We discuss visualization of amplicons on gels, with and without incorporation of fluorescent primers. In more specialized areas, we examine the utility of temperature and denaturing gradient gel electrophoresis to examine responses of predators to prey diversity, and review the potential of quantitative polymerase chain reaction systems to quantify predation. Alternative routes by which prey DNA might get into the guts of a predator (scavenging, secondary predation) are highlighted. We look ahead to new technologies, including microarrays and pyrosequencing, which might one day be applied to this field.
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              Noninvasive genetic tracking of the endangered Pyrenean brown bear population.

              Pyrenean brown bears Ursus arctos are threatened with extinction. Management efforts to preserve this population require a comprehensive knowledge of the number and sex of the remaining individuals and their respective home ranges. This goal has been achieved using a combination of noninvasive genetic sampling of hair and faeces collected in the field and corresponding track size data. Genotypic data were collected at 24 microsatellite loci using a rigorous multiple-tubes approach to avoid genotyping errors associated with low quantities of DNA. Based on field and genetic data, the Pyrenean population was shown to be composed at least of one yearling, three adult males, and one adult female. These data indicate that extinction of the Pyrenean brown bear population is imminent without population augmentation. To preserve the remaining Pyrenean gene pool and increase genetic diversity, we suggest that managers consider population augmentation using only females. This study demonstrates that comprehensive knowledge of endangered small populations of mammals can be obtained using noninvasive genetic sampling.
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                Author and article information

                Contributors
                forgacs1@tamu.edu
                rick_wallen@nps.gov
                aboedeker@cvm.tamu.edu
                jderr@cvm.tamu.edu
                Journal
                BMC Genet
                BMC Genet
                BMC Genetics
                BioMed Central (London )
                1471-2156
                26 February 2019
                26 February 2019
                2019
                : 20
                : 22
                Affiliations
                [1 ]ISNI 0000 0004 4687 2082, GRID grid.264756.4, Interdisciplinary Graduate Program of Genetics, , Texas A&M University, ; College Station, TX 77843 USA
                [2 ]ISNI 0000 0004 4687 2082, GRID grid.264756.4, Department of Veterinary Pathobiology, , Texas A&M University, ; College Station, TX 77843 USA
                [3 ]ISNI 0000 0001 2331 3972, GRID grid.454846.f, National Park Service, Yellowstone National Park, ; Hot Springs, Mammoth, WY 82190 USA
                Author information
                http://orcid.org/0000-0001-9940-5611
                Article
                722
                10.1186/s12863-019-0722-3
                6390568
                30808294
                dd72a3cc-47c7-47b0-afc2-f6f66e10a0ba
                © The Author(s). 2019

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 20 March 2018
                : 8 February 2019
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100007516, National Park Service;
                Award ID: P12AC71337
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2019

                Genetics
                bison,yellowstone national park,fecal dna,microsatellite,str,heterozygosity,allelic dropout
                Genetics
                bison, yellowstone national park, fecal dna, microsatellite, str, heterozygosity, allelic dropout

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