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Assessing the utility of metabarcoding for diet analyses of the omnivorous wild pig (Sus scrofa)
Authors:Michael S Robeson II  Kamil Khanipov  George Golovko  Samantha M Wisely  Michael D White  Michael Bodenchuck  Timothy J Smyser  Yuriy Fofanov  Noah Fierer  Antoinette J Piaggio
Institution:1. Fish and Wildlife Conservation Biology, Colorado State University, Fort Collins, CO, USA;2. USDA, Wildlife Services, National Wildlife Research Center, Wildlife Genetics Lab, Fort Collins, CO, USA;3. Department of Pharmacology, The University of Texas Medical Branch, Galveston, TX, USA;4. Department of Wildlife Ecology and Conservation, USA5 USDA, Wildlife Services, University of Florida, Gainesville, FL, USA;5. Tejon Ranch Conservancy, Frazier Park, CA, USA;6. USDA, APHIS, Wildlife Services, San Antonio, TX, USA;7. Department of Ecology and Evolutionary Biology, Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, CO, USA
Abstract:Wild pigs (Sus scrofa) are an invasive species descended from both domestic swine and Eurasian wild boar that was introduced to North America during the early 1500s. Wild pigs have since become the most abundant free‐ranging exotic ungulate in the United States. Large and ever‐increasing populations of wild pigs negatively impact agriculture, sport hunting, and native ecosystems with costs estimated to exceed $1.5 billion/year within the United States. Wild pigs are recognized as generalist feeders, able to exploit a broad array of locally available food resources, yet their feeding behaviors remain poorly understood as partially digested material is often unidentifiable through traditional stomach content analyses. To overcome the limitation of stomach content analyses, we developed a DNA sequencing‐based protocol to describe the plant and animal diet composition of wild pigs. Additionally, we developed and evaluated blocking primers to reduce the amplification and sequencing of host DNA, thus providing greater returns of sequences from diet items. We demonstrate that the use of blocking primers produces significantly more sequencing reads per sample from diet items, which increases the robustness of ascertaining animal diet composition with molecular tools. Further, we show that the overall plant and animal diet composition is significantly different between the three areas sampled, demonstrating this approach is suitable for describing differences in diet composition among the locations.
Keywords:blocking primer  CO1  diet  feral swine  metabarcoding  trnL
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