Journal of the North Carolina Academy of Science, 131(1), 2015, pp. 15–17

A CAVE , SPHALLOPLANA PERCOECA, (PLATYHELMINTHES, TRICLADIDA, ) FROM GREAT SMOKY MOUNTAINS NATIONAL PARK

BENNY GLASGOW1 and PAULA PIERCE2 1Principal investigator, Park study GRSM-00341, 101 William Street, Vernon, AL 35592 Email: [email protected] 2Excalibur Pathology, Inc., 5830 N Blue Lake Dr., Norman OK 73069 Email: [email protected] Downloaded from http://meridian.allenpress.com/jncas/article-pdf/131/1/15/1818171/2167-5872-131_1_15.pdf by guest on 27 September 2021 Abstract: Five cave collected from Gregory’s Cave, Blount Co., TN, Great Smoky Mountains National Park, were analyzed using stained serial sections and two are identified as Sphalloplana percoeca (Packard 1879). Notes on specimen characteristics and habitat are given, two photographs are provided, and the species’ and distribution are discussed. Key Words: Sphalloplana percoeca; Kenkiidae; Gregory’s Cave.

INTRODUCTION 1978, pers. comm. 2010). Aquatic amphipods may be The Great Smoky Mountains National Park’s ongo- food sources for cave planarians (Reeves 2000; Carpen- ing All Taxa Biodiversity Inventory, to determine ter 1982). Bats, amphibians, amphipods, millipedes, and presence and distribution of Park species or discover insects were observed in the cave during Park study new species, elicits this report on an obligate cave GRSM-00341 collections. Mays (2001) reported amphi- planarian belonging to family Kenkiidae (Hyman 1937). pods, millipedes, and spiders inhabiting the cave. Larvae Cave planarians are not well known and there could be of the Long-tailed Salamander (Eurycea longicauda) new species yet undiscovered. Gregory’s Cave was were reported in the cave’s rimstone pools (Dodd et al. searched to find cave planarians and identify them as 2001). a known or a new species. Cave planarian is a general term most often used to METHODS AND MATERIALS describe troglobitic freshwater of the order Tricladida. Cave planarians of family Kenkiidae have Gregory’s Cave was searched for cave planarians characteristics that may be the result of cave existence, under permits issued for Park study GRSM-00341. including an anterior adhesive organ, lack of pigmen- Specimens were collected from rimstone pools in the tation, and the absence of eyes (Carpenter 1982). Cave cave using handheld pipettes and placed in a container planarians are reported to have low tolerance for with water from the cave. conditions outside of caves and brief exposure to Specimens were removed from the cave, examined sunlight has resulted in writhing, death, and disintegra- with a hand lens, and fixed in Excalibur’s alcoholic Z- tion (Carpenter 1982; McRitchie 1959). Several uses of fixative or 70% ethanol. Specimens were microtomed in the anterior adhesive organ by cave planarians have 5-micron serial sections, stained with hematoxylin-eosin, been noted, including sensing and capture of prey, an and mounted on slides. Prepared slides were examined aid in movement, and grasping the substrate to prevent using compound light microscopes. being washed away by water currents (Hyman 1951; Materials are notes, drawings, photographs, and five Carpenter 1982). sets of serial sections labeled T-15, 2013-1E, 2013-1C, Cave planarians identified as Sphalloplana sp. were 2013-1B, and 2013-1A. All slides are in longitudinal reported in Gregory’s Cave, Blount Co., TN, by John R. sections except for 2013-1E, which is longitudinally and Holsinger in 1964 (Park records, pers. comm. 2010). obliquely sectioned. Slides, notes, drawings, and photo- Reeves (2000) reported Sphalloplana sp. from Gregory’s graphs are archived in Great Smoky Mountains Cave and Whiteoak Blowhole Cave, Blount Co., TN, National Park collections, Gatlinburg, TN. also located in the Park. Species determinations were not made in these reports. The 133-m long cavern of Gregory’s Cave is formed in RESULTS AND DISCUSSION dolomitic rock (Dodd et al. 2001). The cave is habitat Specimens were eyeless, unpigmented white, measured for a diversity of species and is the type locality for the up to 12 mm long when crawling, extended, and were 2 rare cave amphipod, Stygobromus sparsus (Holsinger mm to 3 mm wide. Anterior ends were truncate with 15 16 JOURNAL OF THE NORTH CAROLINA ACADEMY OF SCIENCE 131(1) Downloaded from http://meridian.allenpress.com/jncas/article-pdf/131/1/15/1818171/2167-5872-131_1_15.pdf by guest on 27 September 2021

FIG. 1. Serial section of the copulatory apparatus of Gregory’s Cave specimen 2013-1B showing the entry of the common oviduct into male antrum. Part of penis papilla (PP), male antrum (MA), and entry of common oviduct (EOD). Dorsal side is toward the top and ventral FIG. 2. A section of the copulatory apparatus of specimen 2013- toward the bottom. 1A. Copulatory bursa (CB), entry of sperm duct into the penis bulb (SDE), penis papilla (PP), male antrum (MA), gonopore (GO), and vagina (VG). Dorsal side is at top and ventral at bottom. a central adhesive organ and with auricles projecting anterolaterally. Specimens had a developed post-pha- ryngeal region. Anterior intestinal borders were rounded (Kenk 1977). They are like them in having and without V-shaped appearance. internal pharyngeal muscles in separate layers (Hyman Slides revealed the inner pharyngeal muscles of the 1951) and moderate number of prepharyngeal testicular five specimens were in separate layers, with an inner follicles (Sluys et al. 2009). However, the Kenkiidae differ circular layer covered by a longitudinal layer, and with from Planariidae in having anterior adhesive organs no latticed intermingling of circular and longitudinal (Hyman 1951). The adhesive organ has a glandular lumen muscles. The adhesive organs were weakly developed and can be protruded and withdrawn by the and not pit-like or deeply invaginated. (Hyman 1937). The Turbellarian Taxonomic Database The internal morphology of two planarian specimens web site, turbellaria.umaine.edu, provides more infor- from Gregory’s Cave were examined using stained serial mation about classifications and related species. sections and are identified as Sphalloplana percoeca Sphalloplana percoeca is known from caves of using descriptions by Kenk (1972, 1977) and Hyman Kentucky (Packard 1879), Alabama (Carpenter 1982; (1951). Specimens 2013-1B and 2013-1A (Figs. 1 and 2) Kenk 1977), West Virginia (Kenk 1977), and Tennessee are like descriptions of S. percoeca in the dorsal location (Carpenter 1970, pers. comm. 2015). The current article of the testes; the separate and lateral entry of the sperm identifies the species from a Tennessee cave in Great ducts into the penis bulb; the entry of the common Smoky Mountains National Park. Kenk (1977) states oviduct into the dorsal part of the male atrium at the that all records of S. percoeca from Tennessee need transition of the common atrium (Fig. 1); the common verification. Examination of internal anatomy using oviduct opening is not near the gonopore; the distinctly stained serial sections is required to identify species of voluminous expansion of the posterior bursal duct or Sphalloplana or determine whether specimens are that of vagina; the shape of the papilla; the location of the a new species (Kenk 1972). Carpenter (1970) indicated copulatory bursa (Fig. 2); and the shape of the that he had identified S. percoeca from Indian Cave, copulatory apparatus. White Co., TN., based on live specimens he collected and The three other specimens from Gregory’s Cave have serial sections he made. Carpenter (1982) mentions an incomplete copulatory apparatus and are not morphological examination of specimens from Ken- identified to species. They have an anterior adhesive tucky and Alabama, but does not provide locations from organ and internal pharyngeal muscles in separate Tennessee. McRitchie (1959) reports S. percoeca from layers, identifying them as Sphalloplana sp. They show Mill Creek Cave, Davidson Co., TN, and Herring Cave, no characteristics indicating they are a separate species Rutherford Co., TN, but it was not determined whether from specimens 2013-1A and 2013-1B. specimens were identified using stained serial sections. Recent taxonomic schemes classifies Sphalloplana The authors emphasize that any identification of cave percoeca in family Kenkiidae (Sluys et al. 2009; Tyler planarians based on external appearance should be et al. 2006–2012). The Kenkiidae are similar to family considered as probable but uncertain identification, GLASGOW AND PIERCE: ACAVE PLANARIAN, SPHALLOPLANA PERCOECA 17 even when the investigator is experienced, because many HOLSINGER, J. R. 1978. Systematics of the subterranean amphipod species of cave planarians are externally similar. Kenk Stygobromus (Crangonyctidae), part II: species of the (1972, 1978) and Hyman (1951) both state that Eastern United States. Smithson. Contrib. Zool. 266:144. HYMAN, L. H. 1937. Studies on the morphology, taxonomy, and identification of planarians require examination of their distribution of North American Triclad Turbellaria. VIII. Some internal histology and morphology, which is best done cave planarians of the United States. Trans. Amer. Microsc. using stained serial sections. Kenk (1972) gives a synop- Soc. 56:457–477. sis of techniques used in planarian identification. HYMAN, L. H. 1951. North American triclad turbellaria. XII. Acknowledgments: We thank the staff and partners Synopsis of the known species of fresh-water planarians of North America. Trans. Amer. Microsc. Soc. 70(2):154–167. of the Great Smoky Mountains National Park for their KENK, R. 1972. Freshwater planarians (Turbellaria) of North assistance. Thanks to Discover Life in America staff and America. Biota of Freshwater Ecosystems, I.D. Manual volunteers for their assistance and for providing grants No. 1, Environmental Protection Agency, U.S. Govt. Printing supporting the research. Thanks to Park staff and Office, Washington, ix + 81 pp. Downloaded from http://meridian.allenpress.com/jncas/article-pdf/131/1/15/1818171/2167-5872-131_1_15.pdf by guest on 27 September 2021 volunteers who assisted with collections in Gregory’s KENK, R. 1977. Freshwater Triclads (Turbellaria) of North America Cave. Thanks to Jonathan Mays, Ann Froschauer, Dan IX: the Genus Sphalloplana. Smithson. Contrib. Zool. 246:38. MAYS, J. D. 2001. The Arthropods of Gregory’s cave. ATBI Q. 2(3):9. Nolfi, Jeannie Hilten, Adriean Mayor, and Carole MCRITCHIE, R. G. 1959. The Kenkiidae of the Nashville Area, v + 43 Middleton for their assistance. pp., 10 plates. 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