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Acronyms and Abbreviations Acronyms and Abbreviations AAFB Anderson Air Force Base AAFB-NWF Anderson Air Force Base - Northwest Field AH Apra Harbor BA Barrigada BOLD Barcode of Life Database bp Nucleotide base pairs CATR Cooperative Agreement Technical Representative cm2 Square centimeters CNMI Commonwealth of the Northern Mariana Islands COI Cytochrome Oxidase Subunit I DNA Deoxyribonucleic Acid DoD Department of Defense DON Department of the Navy ESA Endangered Species Act EtOH Ethanol FG Finegayan GPS Global Positioning System HERA Haputo Ecological Reserve Area m Meters m2 Square Meters mm Millimeters mtDNA Mitochondrial DNA NAVFAC Naval Facilities Engineering Command NBGAH Naval Base Guam Apra Harbor NBGTS Naval Base Guam Telecommunications Site NWF Northwest Field PAUP* Phylogenetic Analysis Using Parsimony PCR Polymerase Chain Reaction RT Route 15 SBS Simultaneous Bidirectional Sequencing UoG University of Guam UHM University of Hawaii at Manoa USFWS United States Fish and Wildlife Service FEDERAL CANDIDATE SPECIES SURVEYS ON GUAM Prepared by Daniel P. Lindstrom, Ph.D. & John C. Benedict, Ph.D. University of Guam Unibetsedåt GUAHAN College of Natural and Applied Sciences Prepared For Naval Facilities Engineering Command Marianas January 21, 2014 FEDERAL CANDIDATE SPECIES SURVEYS ON GUAM TABLE OF CONTENTS Acronyms and Abbreviations ....................................................................................... Inside front cover ABSTRACT ................................................................................................................................................ 1 1.0 INTRODUCTION ................................................................................................................................ 3 1.1 General Background ................................................................................................................. 3 1.2 Candidate Species Descriptions ................................................................................................ 4 1.3 Project Objectives ................................................................................................................... 12 2.0 METHODS ......................................................................................................................................... 15 2.1 General Survey Methodology and Timelines ......................................................................... 15 2.2 Butterfly Specific Survey Methodologies and Field Data Collection .................................... 16 2.2.1 Field Sampling for Butterfly Genetics ................................................................... 16 2.2.2 Butterfly Genetic Sequencing and Analysis .......................................................... 17 2.3 Tree Snail Specific Methodologies and Field Data Collection .............................................. 19 2.3.1 Field Sampling for Tree Snail Genetics ................................................................. 20 2.3.2 Snail Genetic Sequencing and Analysis ................................................................ 20 3.0 RESULTS ........................................................................................................................................... 21 3.1 Dry Season General Survey .................................................................................................... 21 3.2 Transect Floral Descriptions With Focus on Known Candidate Species Host ....................... 23 3.3 Wet Season Targeted Search ................................................................................................... 23 3.4 Partulid Home Plants............................................................................................................. 26 3.5 Genetics for Butterflies ........................................................................................................... 26 3.6 Genetics for Tree Snails .......................................................................................................... 27 3.7 Introduced Predators and Ungulates................................... .................................................... 28 3.8 General Survey Results................................................................... ........................................ 28 4.0 DISCUSSION ..................................................................................................................................... 29 5.0 CONCLUSIONS and RECOMMONDATIONS ............................................................................ 32 5.1 General .................................................................................................................................... 32 5.2 Butterflies ............................................................................................................................... 34 5.3 Tree Snails .............................................................................................................................. 35 6.0 LITERATURE CITED ...................................................................................................................... 41 7.0 LIST OF PARTICIPANTS AND PREPARERS ............................................................................ 44 APPENDICES Appendix A: Micronesian tree snail literature review: Partulid tree snails (Partulidae: Stylommatophora) of the Mariana Islands, Micronesia. ......................A1-A31 Appendix B: Candidate butterfly species literature review: The Mariana Eight Spot Butterfly, Hypolimnas octocula marianensis and the Mariana Wandering Butterfly, Vagrans egistina, of the Mariana Islands, Micronesia. ........................ B1-B26 Appendix C: Transect Atlas of GIS/Satellite and Vegetation of all areas surveyed ................... C1-C33 Appendix D: Transect Flora Vegetation Station List ................................................................... D1-D27 Appendix E: Federal Candidate Species Occurrence List ............................................................ E1-E25 Appendix F: Digital Photograph and GIS Database (Supplied on included DVD) List of Figures & Tables Figure Page 1. S. fragilis................................................................................................................................................... 5 2. Dead E. rosea shell................................................................................................................................... 6 3. P. radiolata on Morinda citrifolia............................................................................................................ 7 4. P.gibba on Alocasia macrorriza............................................................................................................... 8 5. H. octocula marianensis adult.................................................................................................................. 9 6. H. octocula marianensis eggs on E. calcareum. .................................................................................... 10 7. H. octocula marianensis chrysalide on P. pedunculata.......................................................................... 10 8. Field team member searching for butterfly eggs, larvae and chrysalides on P.pedunculata. ................ 11 9. M. thompsonii. ........................................................................................................................................ 12 10. Candidate Species Dry Season Survey Areas....................................................................................... 13 11. Candidate SpeciesWet Season Search Areas........................................................................................ 14 12. Partula radiolata on FTA™ card......................................................................................................... 20 13. Tower karst topography. ...................................................................................................................... 22 Table Page 1. Native and Non-Native Plant Species Observed with Partulids............................................................. 26 ABSTRACT Surveys for five Endangered Species Act candidate species were conducted on Guam under a cooperative agreement between the University of Guam and the Department of Navy (DON) (N40192-12-8001). The target species include two nymphalid butterflies, Hypolimnas octocula marianensis and Vagrans egistina, and three partulid snails, Samoana fragilis, Partula radiolata, and Partula gibba. The survey was divided into two temporal components, a dry season (March and April 2013) and wet season (June and July 2013); both were aimed at locating candidate species and recovering DNA for molecular analyses. The dry season survey consisted of 47 transects (>42 km distance) of forested regions of Guam, which were surveyed via visual census methods to determine the presence of candidate species and butterfly host plants. H. octocula marianensis was discovered on two transects, one on Andersen Air Force Base Northwest Field (AAFB-NWF) area, and the other on non- Department of Defense (DoD) lands adjacent to Route 15 on the eastern side of Guam. The host plants of H. octocula marianensis, Procris pedunculata and Elatostema calcareum, were only encountered in heavily forested cliff line/tower karst dominated areas of Guam, which are inaccessible to invasive ungulate herbivores. The host plant of V. egistina, Maytenus thompsonii, was present on multiple transects, but no evidence of V. egistina was
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