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Assessing the Genetic Distinctiveness of Cambarus

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Assessing the Genetic Distinctiveness of Cambarus National Park Service U.S. Department of the Interior Natural Resource Stewardship and Science Assessing the Genetic Distinctiveness of Cambarus (Puncticambarus) acuminatus A Recently Discovered Crayfish in Valley Creek within Valley Forge National Historical Park Natural Resource Technical Report NPS/VAFO/NRTR—2013/752 ON THE COVER Background image: Valley Creek, Valley Forge National Historical Park, near a roadside parking area, ~0.5 mi S. of SR23 on SR252 (40.09158, -75.45633, WGS-84). Inset image: Cambarus (Puncticambarus) cf. acuminatus collected from the same locality as the background image. Photographs by: James W. Fetzner Jr. Assessing the Genetic Distinctiveness of Cambarus (Puncticambarus) acuminatus A Recently Discovered Crayfish in Valley Creek within Valley Forge National Historical Park Natural Resource Technical Report NPS/VAFO/NRTR—2013/752 James W. Fetzner Jr. Section of Invertebrate Zoology Carnegie Museum of Natural History 4400 Forbes Avenue Pittsburgh, PA 15213-4080 ([email protected]) Keith A Crandall Department of Biology Brigham Young University 401 Widtsoe Building Provo, UT 84602 Current Address: Computational Biology Institute The George Washington University Innovation Hall 45085 University Drive Ashburn, VA 20147 May 2013 U.S. Department of the Interior National Park Service Natural Resource Stewardship and Science Fort Collins, Colorado The National Park Service, Natural Resource Stewardship and Science office in Fort Collins, Colorado, publishes a range of reports that address natural resource topics. These reports are of interest and applicability to a broad audience in the National Park Service and others in natural resource management, including scientists, conservation and environmental constituencies, and the public. The Natural Resource Technical Report Series is used to disseminate results of scientific studies in the physical, biological, and social sciences for both the advancement of science and the achievement of the National Park Service mission. The series provides contributors with a forum for displaying comprehensive data that are often deleted from journals because of page limitations. All manuscripts in the series receive the appropriate level of peer review to ensure that the information is scientifically credible, technically accurate, appropriately written for the intended audience, and designed and published in a professional manner. This report received formal peer review by subject-matter experts who were not directly involved in the collection, analysis, or reporting of the data, and whose background and expertise put them on par technically and scientifically with the authors of the information. Views, statements, findings, conclusions, recommendations, and data in this report do not necessarily reflect views and policies of the National Park Service, U.S. Department of the Interior. Mention of trade names or commercial products does not constitute endorsement or recommendation for use by the U.S. Government. This report is available from the Natural Resource Publications Management website (http://www.nature.nps.gov/publications/nrpm/). Please cite this publication as: Fetzner Jr., J. W., and K. A. Crandall. 2013. Assessing the genetic distinctiveness of Cambarus (Puncticambarus) acuminatus: A recently discovered crayfish in Valley Creek within Valley Forge National Historical Park. Natural Resource Technical Report NPS/VAFO/NRTR— 2013/752. National Park Service, Fort Collins, Colorado. NPS 464/120889, May 2013 ii Contents Page Figures............................................................................................................................................. v Tables ............................................................................................................................................ vii Appendices .................................................................................................................................... vii Acknowledgments......................................................................................................................... vii Abstract .......................................................................................................................................... ix Introduction ..................................................................................................................................... 1 Background .............................................................................................................................. 1 Taxonomic Problems ............................................................................................................... 3 Materials and Methods .................................................................................................................... 7 Specimen Acquisition .............................................................................................................. 7 Molecular Analyses ............................................................................................................... 11 DNA Extraction ................................................................................................................ 11 PCR Amplification............................................................................................................ 11 DNA Sequencing .............................................................................................................. 12 Genetic Data Analysis....................................................................................................... 12 Results ........................................................................................................................................... 13 Cambarus acuminatus ........................................................................................................... 13 Pennsylvania Populations ................................................................................................. 13 The C. acuminatus Species Complex................................................................................ 19 Syntype MCZ #3624 ......................................................................................................... 20 Relationships Among Members of the Subgenus Puncticambarus ...................................... 21 iii Contents Page Discussion ..................................................................................................................................... 23 Status of Pennsylvania Populations ....................................................................................... 24 A New Species? ..................................................................................................................... 25 Conclusions and Future Directions ............................................................................................... 27 Literature Cited ............................................................................................................................. 29 iv Figures Page Figure 1. Sampling localities (flag icons) and topography of Valley Creek in Valley Forge National Historical Park (green shaded area). ...................................................................... 2 Figure 2. Illustration of female specimen of Cambarus acuminatus from Plate III, Fig. 5, of Faxon (1885) showing the acuminate rostrum (r) and the cervical (=“branchiostegian”) spines (cs). .................................................................................................... 4 Figure 3. A) Syntype (MCZ 3624) of Cambarus acuminatus from the Museum of Comparative Zoology at Harvard showing the extremely poor condition of the specimen and B) its associated specimen data labels. .................................................................... 5 Figure 4. Gonopod (1st pleopod) of a Form II (juv) male of Cambarus acuminatus. A) illustrations found in Faxon (1885; Plate VII, Figs. 6a, 6a’), B) photographs of the gonopod housed with MCZ specimen #3624 and C) its associated data labels. ............................ 6 Figure 5. Locations where collections were made for A) members of the Cambarus subgenus Puncticambarus and B) for taxa used as outgroups. ....................................................... 8 Figure 6. Map showing locations in Pennsylvania in and around Valley Forge National Historical Park where samples of C. acuminatus were collected for this study (see also Fetzner 2010). ....................................................................................................... 14 Figure 7. Mean estimates of catch per unit effort (± SE) for Pennsylvania populations of C. acuminatus based on collections made in 2005 (Valley Creek) and 2009 (Pickering, Darby, Crum creeks) (see Fetzner 2010). .................................................................. 15 Figure 8. Neighbor-joining tree showing relationships among the 170 unique COI haplotypes found in this study. ..................................................................................................... 17 Figure 9. Maps showing the distribution of mtDNA COI gene haplotype clades that include presumed C. acuminatus samples or related species, as depicted in the phylogeny in Figure 8. .................................................................................................................. 18 Figure 10. Image of an agarose gel showing attempts at amplification of COI gene fragments of 50, 100, and 150 base pairs for three samples
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