BioInvasions Records (2021) Volume 10, Issue 2: 277–286

CORRECTED PROOF

Rapid Communication First Canadian record (Ladysmith Harbour, British Columbia) of the non-native European sponge Hymeniacidon perlevis (Montagu, 1814) (Porifera, Demospongiae)

Rick M. Harbo1,*, Bruce Ott2,*, Henry M. Reiswig3, and Neil McDaniel4 1Research Associate, Invertebrate Zoology, Royal BC Museum, 675 Belleville St, Victoria, British Columbia, V8W 9W2,Canada 24577 West 16th Avenue, Vancouver, BC, V6R 3E8, Canada 3Biology Department, University of Victoria and Royal British Columbia Museum, 675 Belleville St, Victoria, BC, V8W 9W, Canada; deceased 4McDaniel Marine Surveys, Vancouver, BC, Canada Author e-mails: [email protected] (RMH), [email protected] (BO) *Corresponding author

Citation: Harbo RM, Ott B, Reiswig HM, McDaniel N (2021) First Canadian record Abstract (Ladysmith Harbour, British Columbia) of the non-native European sponge The non-native European sponge Hymeniacidon perlevis (Montagu, 1814) (= H. sinapium Hymeniacidon perlevis (Montagu, 1814) de Laubenfels, 1930) is established and abundant in the head of Ladysmith (Porifera, Demospongiae). BioInvasions Harbour, southern British Columbia, Canada, where it was first detected in 2008. In Records 10(2): 277–286, https://doi.org/10. the North Pacific Ocean, it was previously known with certainty from California, 3391/bir.2021.10.2.05 Korea and Japan. It may have been introduced to Ladysmith from Asia between the Received: 6 August 2020 1950s and 1970s with importations of commercial oysters (Crassostrea gigas Accepted: 9 February 2021 (Thunberg, 1793)). Ladysmith Harbour supports some of the warmest marine Published: 31 March 2021 waters of Western Canada; should water temperatures increase in adjacent regions, Handling editor: Thomas Therriault H. perlevis may expand its range and be an important indicator of climate change. Thematic editor: April Blakeslee Key words: Demospongiae, Halichondriidae, introduced species, commercial oyster Copyright: © Harbo et al. vector This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0). Introduction OPEN ACCESS. Abundant growths of an unusual and bright orange intertidal sponge at the head of Ladysmith Harbour, British Columbia, Canada were first noted in May 2008 by the first author. Ladysmith Harbour is a protected bay in the Strait of Georgia on the east coast of Vancouver Island (Figure 1). It is a long, narrow harbour approximately 8 km in length and varying in width from 0.7 to 2 km. At the head of the harbour are extensive mud flats with eight to 15 cm of soft mud, bordered with stretches of rocky shore. The mud flats are exposed at mid-day during the spring and summer low tides, and this heat is transferred to the overlying water during flooding tides. The harbour waters have seasonal temperatures ranging from 5.5 to 23.8 °C making it one of the warmest localities in the Strait of Georgia (Levings et al. 1983; McCallister 1956). Salinities are 13 to 28 psu in the shallow waters at the head of the harbour, influenced at times by three minor creek discharges (McCallister 1956). Ladysmith Harbour is unique in its diverse and abundant intertidal sponges on the mudflats, including Halichondria bowerbanki Burton, 1930,

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 277 First Canadian record of Hymeniacidon perlevis

Figure 1. Ladysmith Harbour, B.C. Hymeniacidon perlevis is limited to the shaded mud flat area at the head of the inlet, Bush Creek and around Page Point.

Haliclona sp. and Lissodendoryx (Lissodendoryx) littoralis Ott, Reiswig, McDaniel & Harbo, 2019. These sponges often form long thin tube-like and furrowed projections that allow them to filter feed in silty conditions. Stehlow et al. (2017) examined several long-term and short-term sediment tolerance mechanisms identified in sponges. While some sponges die from sedimentation from dredging or fishing activities, there are several species that are well adapted to living in environments with high sediment loads. Sponges may employ passive mechanisms, such as growth forms, that limit sediment accumulations inside and on top of the surface. Active mechanisms include the production of mucus, expulsion of particles and temporary cessation of pumping water, and tissue sloughing.

Materials and methods Specimens of sponges were collected intertidally from a rocky shore between May, 2014 and June, 2019 at several sites at the head of Ladysmith Harbour (Figures 1, 2). Most specimens were preserved in 70% isopropyl alcohol. Two samples were preserved in 95% ethanol for future molecular genetic analyses. For material examined, we report museum catalogue number, station number, location, latitude and longitude, depth, date of collection, collector, and number of specimens. Latitude and longitude were taken from Google Earth™ coordinates. For most specimens, colour photographs were taken in situ. Thick sections of specimens were made by excising approximately one cm3 surface blocks and embedding these in 58 °C melting point histological paraffin.

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 278 First Canadian record of Hymeniacidon perlevis

Figure 2. (A) Orange sponge, Hymeniacidon perlevis on intertidal rocks, Page Point, Ladysmith Harbour, B.C.; (B) Hymeniacidon perlevis on wood debris.

After cooling to room temperature, the blocks were trimmed to either vertical or tangential orientation and re-warmed to 40 °C for one-half hour to prevent cracking during sectioning. Warmed tissue blocks were set into a guiding jig and sectioned by hand with a straight razor at thicknesses between 0.1 and 1.0 mm. The sections were de-paraffinized in xylene and, of those, the best sections were mounted on microscope slides in Canada balsam for photography. Tissue-free spicule preparations were made by dissolving small pieces of sponge on microscope slides in sodium hypochlorite, rinsing with distilled water to remove the sodium hypochlorite, then drying and mounting in Canada balsam for spicule counts. Using a compound microscope, we measured the length and width of 100 spicules (unless noted otherwise by n =). We scanned microscope fields for spicules of variable sizes, but ignored obviously ontogenetically young (juvenile) spicules in determining size ranges. We list spicule dimensions as minimum (mean) maximum ± standard deviation. Juvenile spicules were judged to be those that were thinner and shorter than fully developed spicules. In slide preparations examined, juvenile spicules were relatively uncommon (approximately less than 5%). For scanning electron microscopy (SEM), cleaned spicules were either deposited onto membrane filters that were then taped to stubs, or deposited directly on double-sided tape attached to stubs. Preparations were coated with gold-palladium and viewed in a Hitachi S-3500N SEM at the Biology Department, University of Victoria, Victoria, BC Thick skeletal sections for SEM were prepared in a manner similar for SEM spicules after sectioning and dissolving the paraffin. Materials examined have been deposited in the marine invertebrate collections at Royal British Columbia Museum (RBCM), Victoria, BC.

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 279 First Canadian record of Hymeniacidon perlevis

Results Systematics Phylum Porifera Grant, 1836 Class Demospongiae Sollas, 1885 Subclass Heteroscleromorpha Cárdenas, Pérez & Boury-Esnault, 2012 Order Suberitida Chombard & Boury-Esnault, 1999 Family Halichondriidae Gray, 1867 Genus Hymeniacidon Bowerbank, 1858

Species Hymeniacidon perlevis (Montagu, 1814) Synonymy: Hymeniacidon sinapium de Laubenfels, 1930, p. 26 Taxonomy. Turner (2020) has established that the Californian sponge Hymeniacidon sinapium de Laubenfels, 1930 is a junior synonym of the widely distributed European species H. perlevis.

Material Examined (collected by R. Harbo unless otherwise noted) Specimens for SEMs, Thick Sections Page Point, Ladysmith Harbour, BC (49°0′39.99″N; 123°49′14.27″W), Tidal pool, 2019.06.07, n = 2 ( RBCM #019-00134-001 and 019-00134-002, Leg. Harbo).

Other Material Head of Ladysmith Harbour, BC (49°1.484′N; 123°50.889′W) Intertidal: 2014.05.27 n = 1 (RBCM #018-00150-001) and 2014.06.03 n = 1 (RBCM #014-00418-001). Also at RBCM: 014-00417-001, 2014.04.30, Ladysmith Harbour, Wedge Point, 49°1′1.2″N; 123°50′3.048″W 014-00318-001 Melissa Frey collector, 2013.06.27, Ladysmith Harbour, 49°1′26.4″N; 123°50′52.8″W.

External features Thickly encrusting with fistular processes, with a height of 10 to 20 cm, spreading laterally to 2 m over bedrock. Surface superficially smooth, micropapillate, microhispid. Colour in life brilliant orange with a slightly reddish tint. Consistency soft, compressible, easily torn.

Skeleton Ectosome thin fleshy membrane approximately 160 µm thick with a few tangential styles at the base supporting a vertical, vague palisade of styles that penetrate the surface about 20 µm (Figure 3A, B). Exopinacoderm perforated by randomly dispersed ostia. Fistulae 10 to 20 mm long, to 5 mm diameter. Oscula single and at the apex of fistulae, to 2 mm diameter

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 280 First Canadian record of Hymeniacidon perlevis

Figure 3. (A) SEM of surface (494_05top30x), (B) SEM of top (494_07top250x), (C) LM of fistula and osculum, (D) SEM showing sponge area where ectosome scraped off (497_04bottom60x), (E) Styles showing variation in curvature, typical head and point. Length (μm), (F) Small styles.

(Figure 3C). Choanosome spicules in confusion with some wispy plumose tracts extending into fistulae (Figure 3D).

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 281 First Canadian record of Hymeniacidon perlevis

Table 1. Spicule lengths and widths of studied material of Hymeniacidon perlevis. Length (μm) Width (μm) Number Specimen RBCM# min. (mean) max.± SD min. (mean) max. ± SD measured RBCM# 019-00134-001 141 (249) 366 ± 56 3.4 (6.1) 10.2 ± 1.6 100 Harbo 2019.06.07a RBCM# 019-00134-002 146 (266) 370 ± 59 3.9 (8.0) 12.3 ± 2.0 100 Harbo 2019.06.07b BO* 16-01a 2016.08.04 135 (250) 399 ± 62 4.9 (8.4) 13.0 ± 2.0 100 BO* 16-04 2016.08.04 138 (260) 389 ± 65 3.9 (6.9) 12.0 ± 1.8 100 RBCM# 018-00150-001 147 (255) 336 ± 56 3.9 (7.5) 9.1 ± 1.6 25 Harbo 2014.05.27 RBCM# 014-00418-001 147 (271) 368 ± 66 5.2 (7.9) 10.4 ± 1.9 25 Harbo 2014.06.03 * BO = Bruce Ott; RBCM = Royal BC Museum; SD = standard deviation.

Aquiferous canals in both the ectosome and choanosome. Canals in the ectosome 200–300 µm diameter, while those in the choanosome similar to ectosome and much larger forming lacunae 2000–3000 µm diameter (Figure 3D).

Spicules Exclusively smooth styles, moderately curved, moderately long points, thickest at mid shaft; may be subtylostylote and with subterminal heads (Figure 3E, F). Length varies considerably: 135(257) 399 ± 61 μm by 3.5(7.4) 13.0 ± 2.0 µm; n = 450 (Table 1 lists specimens measured). Spicule density is greatest in a layer at the surface 100–200 µm thick. Loose spicules without order are located among the multispicular tracts.

Ecology Hymeniacidon perlevis is limited to approximately 4.5 km of shoreline at the head of Ladysmith Harbour (Figure 1). The sponge grows on a variety of hard substrates, often in the muddy intertidal, including on shells of the introduced oyster Crassostrea gigas (Thunberg, 1793) and the introduced clam Ruditapes philippinarum (A. Adams & Reeve, 1850), as well as on ropes (but not on chemically treated nets). It may completely grow over and engulf the native oyster Ostrea lurida Carpenter, 1864. It is also abundant on intertidal rocks (Figure 2A) and wood debris (Figure 2B) and was abundant in and adjacent to a shallow tidepool at Page Point. The sponge was not found on floating structures or pilings in the harbour but was found on rocks adjacent to a marina. The sponge dies back over the winter and new growth starts again in the spring. The dorid nudibranch Doris montereyensis Cooper, 1863, has been observed on several occasions feeding on intertidal H. perlevis.

Discussion Hymeniacidon perlevis was well established and abundant in Ladysmith Harbour when first observed in 2008. It appears to be tolerant of

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 282 First Canadian record of Hymeniacidon perlevis

environmental extremes, thriving in areas with high summer temperatures and, when exposed to air at low tide, low winter temperatures. It is also tolerant of brackish water as a result of rainfall or outflow from freshwater streams into the intertidal tidepools or depressions that it occupies. Remarkably similar is de Laubenfels (1932) habitat description of this species in southern California: “It is the most abundant sponge on the oyster beds in Newport Bay, where the water often becomes quiet and very warm. It is found in various places up the stream that enters the bay, where it must be in brackish water at low tide during rainy weather. It grows even where the water is almost opaque with suspended mud, where at low tide it is exposed to the very ardent rays of the sun, and where it is always chilled by the very cool waters of the open ocean”. The fistular nature of H. perlevis may allow it to inhabit muddy or silty areas that might otherwise smother non-tubular sponges. The highly visible orange Hymeniacidon has not been reported in Washington State oyster growing areas, confirming how unique Ladysmith Harbour is. The introduced Atlantic Red Beard Sponge, Clathria prolifera (Ellis & Solander, 1786), also tolerant to sedimentation, has been observed in Willapa Bay, Washington, but there are not any records of this sponge in BC.

Distribution and possible introduction vectors Hymeniacidon perlevis was first detected in the Pacific Ocean in southern California, where it was described as a new species, H. sinapium, by de Laubenfels (1930, 1932). It was collected in southern California as early as 1914 (based on Smithsonian Institution collections; P. Fofonoff personal communication to J.T. Carlton). It remained known largely only from southern California until it appeared in the 1990s in estuaries and bays of central California (Tomales Bay and in Monterey Bay’s Elkhorn Slough, where it formed distinctive orange reefs at the latter location) (Wasson et al. 2001; Fuller and Hughey 2013; J.T. Carlton, personal communication, September 2020). Its appearance in central California at the end of the 20th century appears to be strongly related to the climate-induced poleward movements of many Northeast Pacific species (Sorte et al. 2010). A report from the Galapagos Islands (Desqueyroux-Faúndez and Van Soest, 1997) is questionable (R. van Soest, personal communication to J.T. Carlton, 2013). It is also known from Korea and Japan (Sim and Bakus 2008). While Fuller and Hughey (2013) suggested that H. perlevis (as H. sinapium) had been introduced from Japan to southern California, this would appear to be unlikely (Elvin et al. 2018), as H. sinapium was not known in the Western Pacific until the 1950s, after the Korean War (J.T. Carlton personal communication), whereas it was already present in southern California in the early 1900s.

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 283 First Canadian record of Hymeniacidon perlevis

Turner (2020) has established that H. perlevis was present in Europe since the early 1800s and is widespread in the North Atlantic Ocean. The highly disjunct distribution in the Pacific Ocean is thus suggestive of an introduction from the Atlantic, first to southern California and later to Japan and Korea. Given this, the origin of the Ladysmith Harbour population may be California, Asia, or the Atlantic Ocean. As it was presumably transported from the Atlantic to the Pacific as biofouling on vessel hulls, and perhaps from southern California to the Western Pacific during the Korean War by the same means, we cannot dismiss the possibility of vessel-mediated introduction to British Columbia from one of the above regions. Nevertheless, we suggest that a more probable origin of the Ladysmith Harbour population lies in the importation of commercial Pacific oysters (Crassostrea gigas) directly from Japan. Importation of Pacific oysters from Japan into Ladysmith Harbour began in the early 1900s. Quayle (1988) reported that the majority of the Pacific oyster seed imported into British Columbia was from the Miyagi Prefecture, near Sendai on northern Honshu Island. Researchers in Japan (J. Reimer, Yuji Ise, personal communication) confirmed that H. perlevis is common in the Miyagi Prefecture area where oyster seed was collected and then shipped to Washington and BC. The last importation of oyster seed into Ladysmith Harbour was in 1977 (Bourne 1979). However, as noted above, H. perlevis did not occur in the Western Pacific until the 1950s, thus restricting its probable period of introduction to British Columbia to between the 1950s and 1970s. In turn, locations on the east coast of Vancouver Island such as the backwaters of Ladysmith Harbour were not historically well explored. Quayle (1964) and others (e.g. Gillespie 2007) identified a number of non-native species introduced to British Columbia waters, but did not report any non-native sponges. Recent sampling programs for non-indigenous invertebrate species are often targeted on floating structures and settlement plates (e.g. Gartner et al. 2016). Due to the sponge’s restricted distribution in Ladysmith Harbour, limited public access to the foreshore at the head of Ladysmith Harbour, and by the fact the sponge does not grow on floating structures in Ladysmith Harbour, it may have been present but overlooked for several decades. Its absence at this time in the biofouling communities of artificial structures in Ladysmith Harbour is difficult to explain. Hymeniacidon perlevis is eurytopic, occupying an extraordinarily wide range of habitats, including muddy bays and estuaries (as noted above), exposed rocky shores (de Laubenfels 1930) and open ocean kelp forests (Turner 2020) in California. It occurs as a fouling organism on intertidal wood (observations herein). Elvin et al. (2018) report it from the biofouling on the hull of a Japanese vessel that was lost in March 2011 in northern Honshu, as a result of the Japanese Earthquake and Tsunami, and subsequently washed ashore in the

Harbo et al. (2021), BioInvasions Records 10(2): 277–286, https://doi.org/10.3391/bir.2021.10.2.05 284 First Canadian record of Hymeniacidon perlevis

State of Washington in May 2014. And, as noted, it is a presumptive vessel biofouler apparently carried from the Atlantic Ocean to the Pacific Ocean. We suggest that Ladysmith Harbour is an important site to monitor because of the current limited distribution of this sponge. If regional water temperatures increase over time, H. perlevis’ distribution may expand and this highly visible non-native species may thus serve an important indicator of climate change.

Acknowledgements We wish to thank Ms. Heidi Gartner, Invertebrates Collection Manager and Researcher, RBCM for her assistance with cataloguing and accessioning specimens, Dr. Henry Choong, Curator of Invertebrate Zoology and Leah Best, Head of Knowledge, RBCM, for making facilities available at RBCM and funding publication costs, Rob Waters for assisting with collection of sponges from Ladysmith Harbour, Dr. Brent Gowan for help with SEMs and Terry Zlot, The Print Lab, Victoria, BC. We would like to thank Doug and Peggy Kolosoff, Bill Cogswell, Pamela Walker, Ross Shepherd, and Warren Johnny, Stz’uminus First Nations for access to field sites. A big thank you to J.T. Carlton and an anonymous reviewer who provided comments that helped to improve the manuscript. Most of all, we owe a debt of gratitude to the late Dr. William C. Austin and Dr. Henry M. Reiswig for their passion for sponges and their extensive collections and biogeographic data which were drawn upon for development of this paper. All material was collected under appropriate collection permits and approved ethic guidelines.

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