Aquatic Invasions (2018) Volume 13, Issue 1: 163–171 DOI: https://doi.org/10.3391/ai.2018.13.1.12 © 2018 The Author(s). Journal compilation © 2018 REABIC Special Issue: Transoceanic Dispersal of Marine Life from Japan to North America and the Hawaiian Islands as a Result of the Japanese Earthquake and Tsunami of 2011 Research Article Bugula tsunamiensis n. sp. (Bryozoa, Cheilostomata, Bugulidae) from Japanese tsunami marine debris landed in the Hawaiian Archipelago and the Pacific Coast of the USA Megan I. McCuller1,3,*, James T. Carlton1 and Jonathan B. Geller2 1Maritime Studies Program, Williams College-Mystic Seaport, 75 Greenmanville Avenue, Mystic, Connecticut 06355, USA 2Moss Landing Marine Laboratories, 8272 Moss Landing Rd, Moss Landing, California 95039, USA 3Current address: Southern Maine Community College, South Portland, Maine 04106, USA Author e-mails: [email protected] (MIM), [email protected] (JTC), [email protected] (JBG) *Corresponding author Received: 11 April 2017 / Accepted: 6 November 2017 / Published online: 15 February 2018 Handling editor: Amy Fowler Co-Editors’ Note: This is one of the papers from the special issue of Aquatic Invasions on “Transoceanic Dispersal of Marine Life from Japan to North America and the Hawaiian Islands as a Result of the Japanese Earthquake and Tsunami of 2011." The special issue was supported by funding provided by the Ministry of the Environment (MOE) of the Government of Japan through the North Pacific Marine Science Organization (PICES). Abstract A new species of the cheilostome bryozoan genus Bugula Oken, 1815, Bugula tsunamiensis, is described from Japan, having rafted across the North Pacific Ocean on numerous objects released into the ocean by the 2011 Great East Japan Earthquake and Tsunami, and landing in the Hawaiian Islands and on the Pacific Coast of the United States. This is the second species of the Bugula uniserialis Hincks, 1884 group to be reported from Japan. We elevate the Japanese species Bugula scaphoides constricta Yanagi and Okada, 1918 to full species status, B. constricta, based upon distinctions from the stem species. We suggest that Bugula uniserialis reported from the Galapagos Islands is an undescribed species. Key words: ocean rafting, North Pacific, Bugula uniserialis, Bugula constricta, Bugula scaphoides, Bugula pedunculata Introduction include: the resurrection of the genera Bugulina Gray, 1848 and Crisularia Gray, 1848; the description Bryozoans are a diverse group of filter-feeding orga- of the new genus Virididentula Fehlauer-Ale, Winston, nisms well represented in a wide variety of marine Tilbrook, Nascimento and Vieira, 2015; and the res- environments and habitats, and capable of taking triction of the species assigned to Bugula Oken, 1815 advantage of oceanic rafting objects (Winston 1982; with a redefinition of that genus (Fehlauer-Ale et al. Thiel and Gutow 2005; Goldstein et al. 2014). Arbo- 2015). Characteristics shared by all Bugula species rescent forms appear to be particularly well-adapted include a lack of spines on the ancestrula and distal to life on potentially space-limited rafting substrates portion of autozooids, and an ooecium with both an (McCuller and Carlton 2018). ento- and ectooecium that is attached to the inner The Bugulidae (Cheilostomata) are a widespread distal portion of zooids (Fehlauer-Ale et al. 2015). group of arborescent bryozoans that have recently Fehlauer-Ale et al. (2015) distinguish two morpholo- undergone substantial taxonomic revision. Changes gically defined groups within Bugula sensu stricto: 163 M.I. McCuller et al. the Bugula neritina (Linnaeus, 1758)–B. minima voucher collections. A number of colony fragments Waters, 1909 group; and the Bugula uniserialis Hincks, from BF samples intercepted in Hawaii, Oregon, and 1884 group. The latter has a branching pattern with Washington were photographed with a Leica EZ4 long proximal portions of zooids and thus appears to HD (Leica Microsystems, Wetzlar, Germany) and have a uniserial growth pattern. LAS EZ (Leica Microsystems, Wetzlar, Germany) A number of species within the Bugulidae have imaging software, and measured using Fiji (Schindelin been recorded from Japan. Species in the Bugula et al. 2012). For scanning electron microscopy (SEM), neritina-minima group include: B. minima (Okada and specimens were cleaned in sodium hypochlorite solu- Mawatari 1938); B. neritina (Okada 1929; Mawatari tion, rinsed in tap water, and then replaced in ethanol 1981); B. subglobosa Harmer, 1926 (Mawatari 1963), to prevent degradation during transport. Samples were and B. vectifera Harmer, 1926 (Okada and Mawatari then air dried and coated with Au-Pd using an Anatech 1938). The Bugula uniserialis group is represented USA Hummer 6.6 Sputtering System (Anatech, Hay- by only one species, B. scaphoides constricta Yanagi ward, California USA) at 15mA and viewed under a and Okada, 1918. Species of Bugulina in Japan JEOL JSM-7100FLV field emission SEM (JEOL USA include B. californica (Robertson, 1905) (Okada and Inc., Peabody, Massachusetts USA) at 5.0kV accele- Mawatari 1938; Mawatari 1981; Nandakumar et al. rating voltage. Images were retained as TIFF files. 1993); B. flabellata (Thompson in Gray, 1848) Specimens of Bugula from JTMD-BF-339 (Table (Mawatari 1981), and B. stolonifera (Ryland, 1960) S1) were removed from storage in 95% ethanol and (Scholz et al. 2003). At least three species of Bugula rinsed in distilled water. DNA was then extracted that are considered incertae sedis were reported in from ~25 mg subsamples of colonies using the DNEasy Japan: B. hexacantha Ortmann, 1890, B. lophodendron blood and tissue kit (Qiagen, Venlo, Netherlands; Ortmann, 1890, and B. umbelliformis Yanagi and Catalog No. 69504) following the manufacturer’s Okada, 1918 (Okada and Mawatari 1938) Other protocol. A 313 bp fragment of the cytochrome c species within the Bugulidae include: Crisularia oxidase subunit I (COI) mitochondrial gene was pacifica (Robertson, 1905) (Grischenko et al. 2007); amplified by polymerase chain reaction (PCR) using Halophila johnstoniae Gray, 1843 (Ortmann 1890); primers described by Geller et al. (2013) and Leray Dendrobeania japonica (Ortmann, 1890), and et al. (2013), modified to contain T7 and T3 sequencing Viridentula dentata (Lamouroux, 1816) (Okada and primer sequences (underlined): T3_intLCO (5'- AAT Mawatari 1937; Mawatari 1987). TAA CCC TCA CTA AAG GGG GWA CWG GWT Bugula species that are widely distributed and GAA CWG TWT AYC CYC C -3') and T7_jgHCO that have been previously recorded as rafting include (5'- TAA TAC GAC TCA CTA TAG GGT AIA Bugulina flabellata, Bugula neritina, and Bugula CYT CIG GRT GIC CRA ARA AYC A -3'). Each minima (Stevens et al. 1996; Winston et al. 1997; PCR reaction consisted of 25 µL of Green GoTaq Astudillo et al. 2009). None of these species are in (Promega, Madison, Wisconsin, USA; Catalog No. the B. uniserialis group. PRM7123) master mix (400 μM dATP, 400 μM We report here a new species of Bugula within dGTP, 400 μM dCTP, 400 μM dTTP, and 3 mM the B. uniserialis group occurring on objects released MgCl2), 0.2 µM of each primer, 0.2 mg/ml BSA, and into the North Pacific Ocean in March 2011 due to water to a volume of 50 µL. The PCR program was the Great East Japan Earthquake and Tsunami. 3 min at 94 °C followed by 30 cycles of 94 °C for These objects drifted across the ocean to the Central 60 sec, 47 °C for 45 sec, and 72 °C for 90 sec. PCR Pacific (Hawaiian Archipelago) and the Pacific coast products were purified using Agencourt AMPure beads of the USA. (Beckman Coulter, Brea, California USA; Catalog No. A63880) according to the manufacturer’s protocol Materials and methods and submitted to Elim Biopharmaceuticals (Hayward, California, USA) for dideoxy chain termination Bryozoan samples were obtained from JTMD sequencing using T7 and T3 primers. Forward and (Japanese Tsunami Marine Debris) objects (see Carlton reverse sequences were assembled and primer et al. 2017) landing between 2012 and 2016 in North sequencers removed using Geneious 10.1 (Biomatters, America and the Hawaiian Islands. Each object was Auckland, New Zealand). Novel sequences were aligned assigned a unique identification number preceded by to representative COI from Bugula species found in JTMD-BF (BioFouling-number) (Supplementary material Genbank in March 2017, and phylogenetic trees Table S1). generated using FastTree 2.1.5 (Price et al. 2010) Bryozoan specimens (received dried or in 95% etha- using the GTR substitution model. Genbank voucher nol) were loose in submitted samples or were removed specimens from JTMD-BF-339 are deposited at the from their substrate with a scalpel and placed in Royal British Columbia Museum, Victoria, Canada. 164 A new bryozoan species from tsunami debris (Figure 3) of B. tsunamiensis, potentially due to the Results different routes and thus the broad sweep of environ- Class Gymnolaemata Allman, 1856 mental conditions experienced, that their debris items Order Cheilostomata Busk, 1852 took across the North Pacific Ocean. For example, Family Bugulidae Gray, 1848 avicularia length, zooid length and frontal membrane Genus Bugula Oken, 1815 length were larger on average for specimens that arrived on objects intercepted on the Pacific Coast of Bugula tsunamiensis n. sp. the United States, and smaller for specimens on objects (Figures 1–3) that landed in Hawaii (Figure 3). Bugula tsunamiensis Material. JTMD-BF-23, 131, 134, 168, 196, 210, also demonstrates the widest known range of avicularia 212, 223, 226, 240, 241, 250, 253, 254, 264, 290, size of any Bugula species. Bugula tsunamiensis 304, 339, 352, 353, 356, 390, 398, 402, 410, 413, appears to follow the “temperature-size rule” (see 414, 415, 471, 526, 530, 531, 532, 533, 555, 578, Atkinson 1994) which has been found to be closely 626, 652, 668, 669, 670, 671, 672 (Table S1). These followed by cheilostome bryozoans (O’Dea and rafted objects include material washed ashore from Okamura 1999; Okamura et al. 2011). Japan in Washington, Oregon, California, and Hawaii, Phylogenetics.