Calcareous Sponges from the French Polynesia (Porifera: Calcarea)

Home , Clathrina, Clathrinidae, Ernstia

Zootaxa 4748 (2): 261–295 ISSN 1175-5326 (print edition) https://www.mapress.com/j/zt/ Article ZOOTAXA Copyright © 2020 Magnolia Press ISSN 1175-5334 (online edition) https://doi.org/10.11646/zootaxa.4748.2.3 http://zoobank.org/urn:lsid:zoobank.org:pub:661CD94A-130B-4BD8-B201-28B079815618

Calcareous sponges from the French Polynesia (Porifera: Calcarea)

MICHELLE KLAUTAU1,5, MATHEUS VIEIRA LOPES1, BRUNA GUARABYRA1, ERIC FOLCHER2, MERRICK EKINS3 & CÉCILE DEBITUS4 1Universidade Federal do Rio de Janeiro, Instituto de Biologia, Departamento de Zoologia, Av. Carlos Chagas Filho, 373, CEP 21941- 902, Rio de Janeiro, RJ, Brasil. 2IRD, centre de Nouméa, SEOH, BPA5, 98713 Nouméa cedex, New Caledonia 3Queensland museum, PO Box 3300, South Brisbane BC, Queensland 4101, Australia 4IRD-CNRS-UBO-IFREMER, UMR6539 LEMAR, IUEM, rue Dumont d’Urville, F29280 Plouzané, France 5Corresponding author. E-mail: [email protected]

Abstract

Although the French Polynesian reefs are among the most well studied reefs of the world, sponges are still poorly known, with only 199 species or OTUs of sponges having been described from French Polynesia, 167 at an OTU level and 32 at a species level. From those 199 species, just five are calcareous sponges. As it is possible that this number is underestimated, the aim of the present work was to study the diversity of calcareous sponges from French Polynesia. Hence, different French Polynesian archipelagos were surveyed by SCUBA from 3 to 60 m of depth. Identifications were performed using morphological and molecular (ITS and C-LSU) tools. We found a total of nine species of Calcarea, comprising five different genera. Five species are new to science: Clathrina fakaravae sp. nov., Clathrina huahineae sp. nov., Ernstia variabilis sp. nov., Leucascus digitiformis sp. nov., and Leucandra tahuatae sp. nov. With the present work, the number of identified sponges from French Polynesia at a species level increased from 32 to 41. The only calcareous sponge previously known from French Polynesia that was recollected by our group was Leucetta chagosensis. Our results suggest that the Eastern Indo-Pacific Realm shows more affinity with the Central and the Western Indo-Pacific Realms. Four species supported these affinities: Ascandra cf. crewsi, previously known only from Papua New Guinea, Leucascus simplex from South Australia, and Leucetta chagosensis and L. microraphis, both widespread species in the Indo-Pacific. These two Leucetta species, however, most likely represent species complexes. Once again the molecular markers ITS and C-LSU helped in the identification of calcareous sponges, showing how important is an integrative taxonomy. Although our work has increased in 250% (6 spp to 15 spp) the diversity of calcareous sponges in French Polynesia, it is most possible that this number is still underestimated.

Key words: biodiversity, Clathrina fakaravae sp. nov., Clathrina huahineae sp. nov., Ernstia variabilis sp. nov., Leucascus digitiformis sp. nov., Leucandra tahuatae sp. nov., ITS, C-LSU

Introduction

French Polynesia is a group of 118 islands and several islets and motus around atolls in the Pacific Ocean, occupy- ing a marine area of 2,500 km2. The islands are grouped in six archipelagos: Marquesas Islands, Society Islands, Tuamotu Archipelago, Gambier Islands, and Austral Islands.These archipelagos are part of two provinces of the Eastern Indo-Pacific Realm, the Southeast Polynesia Province and the Marquesas Province. The French Polynesia reefs are among the most well studied reefs of the world (Salvat et al. 2008), nonetheless, sponges are still poorly known (Kelly-Borges & Valentine 1995; Hall et al. 2013). Currently, only 26 species or OTUs of sponges are known from French Polynesia, five of them being calcareous sponges, i.e., sponges whose skeleton has calcium carbonate spicules: Lelapiella sphaerulifera Vacelet, 1977, Lepidoleucon inflatum Vacelet, 1967, Plectroninia radiata Vacelet, 1967, and Murrayona phanolepis Kirkpatrick, 1910 from Moorea (Society Is- lands Archipelago) and Takapoto (Tuamotu Archipelago) (Vacelet 1977), and Leucetta cf. chagosensis Dendy, 1913 from Moorea (Society Islands Archipelago) (Wörheide et al. 2002; Hall et al. 2013) and Rangiroa (Tuamotu Archi- pelago) (Wörheide et al. 2002). It is possible that this number is underestimated, as no expeditions were dedicated

Accepted by J. Hooper: 24 Jan. 2020; published: 6 Mar. 2020 261 to study calcareous sponges before. Therefore, the aim of the present work was to study the diversity of calcareous sponges from French Polynesia.

Materials and methods

Specimens collection In the present study, calcareous sponges collected during the surveys of the different French Polynesian archipelagos: Society Islands (Mehetia, Tahiti, Moorea, Tetiaroa, Huahine, Raiatea and Bora-Bora), Marquesas Islands (Nuku Hiva, Ua Pou, Ua Huka, Tahuata, Hiva Oa, Fatu Hiva), Tuamotu-Gambier Islands (Anuanuraro, Hereher- etue, Tematangi, Tureia, Nukutavake, Tureia, Hao, Amanu, Marokau, Raroia, Makemo, Fakarava, Toau, Takaroa, Rangiroa, Tikehau, Makatea), and Austral Islands (Marotiri, Rapa, Raivavae, Tubuai, Rurutu, Rimatara and Maria Islands) (Debitus, 2009, 2011, 2013a, b) (Fig 1). The collections were performed by SCUBA from 3 to 60 m of depth. When possible, the sponges were photographed in situ and then fixed and preserved in ethanol 93%. All the specimens are deposited at the Museum National d’Histoire naturelle (Paris).

FIGURE 1. Map of the study area. A—French Polynesia. B—Studied islands.

Morphological analysis The sponges were analysed under a stereomicroscope and slides to analyse skeletal organisation and spicules followed standard procedures (Wörheide & Hooper 1999; Klautau & Valentine 2003). Spicule measurements were taken with an ocular micrometer and are presented in tabular form, featuring length and width (minimum [min], mean, standard deviation [SD] and maximum [max]). We measured a total of 20 spicules of each category, always searching for the apparent smallest and the biggest spicules and measuring randomly 18 spicules. Skeleton and spicule photographs were taken with a digital Canon camera coupled to a Zeiss Axioscop mi- croscope. Scanning electron microscopy (SEM) micrographs were taken at the Biology Institute of the UFRJ on a JSM-6510 SEM equipment. Spicule preparations for SEM followed Azevedo et al. (2015).

Molecular analysis The genomic DNA was extracted by the guanidine/phenol-chloroform method (Sambrook, Fritsch & Maniatis 1989) or with a QIAamp DNA MiniKit (Qiagen) and stored at –20°C until amplification. For Calcinea, primers situated in the adjacent 18S and 28S regions were used to amplify and sequence the internal transcribed spacer (ITS) region, containing ITS1, the 5.8S rDNA and ITS2. The primers used were: fwd: 5’-TCATTTAGAGGAAGTAAAAGTCG- 3’ and rv: 5’-GTTAGTTTCTTTTCCTCCGCTT-3’ (Lôbo-Hajdu et al. 2004). For Calcaronea, we amplified the C- LSU region (Voigt & Wörheide 2016), using the primers fwd: 5′-GAAAAGCACTTTGAAAAGAGA-3′ (Voigt & Wörheide 2016) and rv: 5′-TCCGTGTTTCAAGACGGG-3′ (Chombard, Boury-Esnault & Tillier 1998). The PCR mix included 1X buffer (5X GoTaq Green Reaction Buffer Flexi, PROMEGA), 0.2 mM dNTP, 0.5

262 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. μg/μL bovine serum albumin (BSA), 2.5 mM MgCl2, 0.33 μM of each primer, one unit of Taq DNA polymerase (Fermentas or PROMEGA) and 1 μL of DNA in a final volume of 15 μL. The PCR amplification consisted of one cycle of 4 min at 94 °C, 35 cycles of 1 min at 92 °C, 1 min at 48° or 50 °C and 1 min at 72 °C, followed by a final cycle of 6 min at 72 °C. Forward and reverse strands were sequenced in an ABI 3500 (Applied Biosystems) at the Biology Institute of the Universidade Federal do Riode Janeiro (UFRJ). All the sequences obtained were analysed and edited in the program GeneStudio and BLAST searches (http://www.ncbi.nlm.nih.gov/blast/) were performed to confirm their biological source. Sequences retrieved from the Genbank database were also used and are listed in Table 1 with those generated in this study. Sequences were aligned through the MAFFT v.7 online platform (Katoh & Standley 2013) with the strategy Q-INS-i (Katoh & Toh 2008), taking into consideration the secondary structure of ribosomal DNA. For the other parameters we chose the default option. The final alignment of the ITS sequences was 1230 bp, including gaps. A total of 480 conserved sites, 571 variable sites, and 138 singletons were retrieved. For the C-LSU of Calcinea, the final alignment presented 427 bp, including gaps, and we retrived 287 conserved sites, 139 variable sites, and 29 singletons. The alignment of the C-LSU of Calcaronea had 404 bp, including gaps. A total of 271 conserved sites, 129 variable sites, and 31 singletons were retrieved. When a sequence from GenBank was longer than the final alignments size mentioned above, it was shortened. The nucleotide substitution model that best fit each alignment was indicated by the Bayesian Information Cri- terion in MEGA 6 (Nei & Kumar 2000; Tamura et al. 2013): GTR + G + I for Calcinea (ITS), T92 + G for Calcinea (LSU), and TN93 + G for Calcaronea. A Maximum Likelihood (ML) tree was built in MEGA 6 using an initial NJ tree (BIONJ) and 1000 pseudo-replicates bootstrap. Bayesian analises were performed in MrBayes 3.1.2. (Huelsen- beck & Ronquist 2001; Ronquist & Huelsenbeck 2003) under 106 generations and a 25% burn in, resulting in a consensus tree of majority. The posterior probability values are shown in the ML tree. The trees of Calcinea were midpoint-rooted, because the only apropriate outgroup was Calcaronea and calcaro- nean sequences do not align properly with Calcinea. For the Calcaronea tree, as we had only a sequence of Leucan- dra in our ingroup, we could use Leucosolenia to root the tree. In order to estimate the genetic intraspecific and interspecific variability, we calculated the uncorrected p-distance in MEGA 6.

TABLE 1. Specimens included in the phylogenetic analyses. (*) DNA sequences generated in the present work. H = holotype, P = paratype. Species Locality Voucher Number GenBank GenBank accession accession number number (ITS) (C-LSU) Subclass Calcinea Ascoleucetta compressa Australia UFRJPOR 7109 - MG595265 Clathrina antofagastensis Peru MNRJ 13674 KF002723 - Clathrina aphrodita Peru MNRJ 12994 KC985138 - Clathrina aurea Brazil MNRJ 8998 HQ588968 - Clathrina blanca Adriatic PMR 14307 KC479087 - Clathrina clathrus Mediterranean UFRJPOR 6315 HQ588974 - Clathrina conifera Brazil MNRJ 8991 HQ588959 - Clathrina coriacea Norway UFRJPOR 6330 HQ588986 - Clathrina curacaoensis Curaçao UFRJPOR6734 MF472607 MF472607 Clathrina cylindractina Brazil UFRJPOR 5206 HQ588979 - Clathrina fjordica Chile MNRJ 8143 HQ588984 - Clathrina hispanica Mediterranean UFRJPOR 6305 KC843432 - Clathrina huahineae sp. nov. French Polynesia UFRJPOR 1600 KC843438 - Clathrina huahineae sp. nov. French Polynesia UFRJPOR 6461 (H) KC843439 - ...... continued on the next page

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 263 TABLE 1. (Continued) Species Locality Voucher Number GenBank GenBank accession accession number number (ITS) (C-LSU) Clathrina insularis Brazil UFRJPOR 6536 (P) KX548922 - Clathrina lacunosa Norway UFRJPOR 6334 HQ588991 - Clathrina lutea Brazil UFRJPOR 5172 (H) HQ588961 - Clathrina luteoculcitella Australia QMG 313684 HQ588989 - Clathrina mutabilis Brazil UFRJPOR 6525 MN422244 - Clathrina mutabilis Brazil UFRJPOR 6528 (P) KX548926 - Clathrina mutabilis Brazil UFRJPOR 6539 MN422245 - Clathrina mutabilis Caribbean UFRJPOR 6741 KC843437 - Clathrina nuroensis Peru MNRJ 13032 KC985136 - Clathrina peruana Peru MNRJ 12839 KC985135 - Clathrina primordialis Adriatic PMR 14305 KC479086 - Clathrina ramosa Chile MNRJ 10313 HQ588990 - Clathrina rotundata Red Sea SMF 11636 KY711435 - Clathrina rowi Red Sea SMF 11629 KY366402 - Clathrina rubra Adriatic PMR 14306 KC479088 - Clathrina sinusarabica Red Sea GW 3143 KY366405 KY366405 Clathrina smaragda Florida UFRJPOR 8359 (H) MG017974 - Clathrina wistariensis Australia QMG 313663 HQ588987 - Ernstia adunca Martinique UFRJPOR 7672 - MG6446123 Ernstia arabica Red Sea SNSB BSPG GW1130 - KY366358 Ernstia arabica Red Sea SMF11627 KY366406 - Ernstia citrea Brazil UFRJPOR 6621 (H) KC843433 - Ernstia citrea Brazil UFRJPOR 6649 - Ernstia klautauae Indonesia RMNHPOR 9341 (H) - MF686062 Ernstia klautauae Indonesia ZMAPOR 08390 (P) KC843451 - Ernstia naturalis Indonesia RMNHPOR 11719 - MF872758 Ernstia aff. naturalis Mayotte RMNHPOR 8444 - MF872757 Ernstia pyrum Aquarium in Russia ZIN 11879 (H) MK617946 - Ernstia rocasensis Brazil UFRJPOR 6664 KX548928 - Ernstia solaris Brazil UFRJPOR 6538 KX548915 - Ernstia tetractina Brazil UFRJPOR 5183 HQ589000 HQ589021 Ernstia variabilis sp. nov.* French Polynesia UFRJPOR 8962 MN422247 MN422250 Ernstia variabilis sp. nov.* French Polynesia UFRJPOR 8963 (H) MN422248 MN422251 Ernstia variabilis sp. nov.* French Polynesia UFRJPOR 8964 MN422249 MN422252 Leucascus flavus Indonesia RMNHPOR 2279 - MF686080 Leucascus simplex* French Polynesia UFRJPOR 6451 - MN422254 Leucascus simplex* French Polynesia UFRJPOR 6456 - MN422255 Leucascus sp. Australia QM G316051 - JQ272305 Leucetta antarctica Antarctic MNRJ 13798 KC849700 - Leucetta avocado Mauritius NCI 108 - KC869542 Leucetta chagosensis French Polynesia BMOO16210 KC843454 - ...... continued on the next page

264 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. TABLE 1. (Continued) Species Locality Voucher Number GenBank GenBank accession accession number number (ITS) (C-LSU) Leucetta chagosensis Australia - - AY563543 Leucetta chagosensis Australia QM G316279 - JQ272296 Leucetta chagosensis Indonesia ZMAPOR 13283 - MF686085 (ex-Ascoleucetta sagittata) Leucetta chagosensis Rodrigues RMNHPOR 11657_1 - MF872790 Leucetta chagosensis Rodrigues RMNHPOR 11658 - MF872791 Leucetta chagosensis* French Polynesia UFRJPOR 6455 - MN422257 Leucetta delicata Antarctic SMF 11868 KC874654 - Leucetta floridana Brazil UFRJPOR 4703 EU781979 - Leucetta floridana Panama P10x2 - KC869538 Leucetta foliata Australia UFRJPOR 7149 - KX499451 Leucetta giribeti Antarctic NHMUK:2017.1.19.1 KY670632 KY670632 Leucetta microraphis Australia QM G313659 AF479061 JQ272297 Leucetta microraphis* French Polynesia UFRJPOR 6450 MN422246 MN422256 Leucetta microraphis* French Polynesia UFRJPOR 6457 - MN422258 Leucetta microraphis* French Polynesia UFRJPOR 6459 - MN422259 Leucetta microraphis Australia QM G313691 - JQ272298 (ex-Leucetta sp.) Leucetta microraphis Mayotte RMNHPOR 8318 - MF872796 Leucetta microraphis Mayotte RMNHPOR 8341 - MF872795 Leucetta microraphis Madagascar RMNHPOR 8717 - MF872797 Leucetta microraphis Indonesia RMNHPOR 6610 - MF686087 Leucetta microraphis Red Sea SNSBBSPG GW 3163 KY366374 Leucetta microraphis ? QM G315140 AJ633871 - Leucetta microraphis Red Sea SNSBBSPG GW 3164 KY36640 KY366375 Leucetta microraphis Red Sea SNSBBSPG GW 3196 KY366403 - Leucetta potiguar Brazil MNRJ 8474 EU781981 Leucetta purpurea Australia UFRJPOR 7255 - KX499450 Leucetta cf. pyriformis Antarctic MNRJ 13843 KC843457 - Leucetta sulcata Rodrigues RMNH11639_1 - MF872798 Leucetta sulcata Rodrigues RMNH11639_2 - MF872799 Leucetta sulcata Rodrigues RMNH11643 - MF872800 Leucetta sulcata Rodrigues RMNH11645 - MF872801 Leucetta villosa Maldives QM G313662 - JQ272295 Leucettusa haeckeliana Tasmania QM G323232 - JQ272300 Leucettusa imperfecta Tasmania QM G323283 - JQ272299 Leucettusa nuda Chile MNRJ 10804 KC843453 - Leucettusa sp. New Zealand OCDN6676-Q KC843458 - Pericharax carteri Australia QM G316099 AJ633882 - Pericharax crypta Australia UFRJPOR 7129 - KX499452 Pericharax orientalis Australia NCI 400 - KC869505 ...... continued on the next page

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 265 TABLE 1. (Continued) Species Locality Voucher Number GenBank GenBank accession accession number number (ITS) (C-LSU) Pericharax vallii Australia UFRJPOR 7126 - KX499449 Subclass Calcaronea Breitfussia schulzei West Greenland FB61 - MH385228 Grantia compressa Roscoff, France (C38) SA80 - MH385242 Leucandra ananas Norway, Trondheimsfjord, SA136 - MH385249 2016032 No.16 Leucandra aspera Marseille, France (C62bis) ? - AY563535 Leucandra falakra Adriatic Sea PMR 13748 - KT447560 Leucandra cf. gausapata Antarctic HT15 - MH385252 Leucandra mozambiquensis Mozambique ZMA.POR.22408 - MF872766 Leucandra nicolae Australia QM G313672 (H) - JQ272268 Leucandra penicillata West Greenland FB35 - MH385256 Leucandra pilula Seychelles ZMA.POR.10528 (H) - MF872767 Leucandra sp. Pacific, Coral Sea, Osprey Reef QM G316285 - JQ272265 Leucandra tahuatae sp. nov.* French Polynesia UFRJPOR 6454 - MN422253 Leucandrilla sp. Galapagos RMNH.POR.11520 - MF872768 Leucilla antillana Curaçao UFRJPOR 6768 (H) - MF472615 Leucilla micropilosa Curaçao UFRJPOR 6755 (H) - MF472621 Leucosolenia botryoides ? SA60 - MH385257 Leucosolenia complicata ? AA09 - MH385268 Leucosolenia cf. corallorrhiza ? FB14 - MH385258 Leucosolenia cf. variabilis ? FB12 - MH385263 Leucosolenia sp. 1 ? FB81 - MH385272 Paraleucilla dalmatica Adriatic Sea PMR 13747 - KT447565 Paraleucilla erpenbecki Mozambique ZMA.POR.22409C (P) - MF872728 Paraleucilla magna South coast of Portugal PT06 - MH385279 Paraleucilla sp.1 South Atlantic, St. Helena SA83 - MH385283 Paraleucilla sp.2 South Atlantic, St. Helena SA85 - MH385284 Sycon caminatum MAD.050202. Madeira Isl. PT02 - MH385301 Madeira Sycon villosum ZMBN 90325, Norway, SA24 - MH385314 Moøyastaken, 26.07.07 Ute glabra Portugal PT16 - MH385315 Ute gladiata Norway, Korsfjord HT47 - MH385316

Results

Systematic Index

Class CALCAREA Bowerbank, 1862 Subclass CALCINEA Bidder, 1898 Order CLATHRINIDA Hartman, 1958

266 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. Family LEUCALTIDAE Dendy & Row, 1913 Genus Ascandra Haeckel, 1872 Ascandra cf. crewsi Van Soest & De Voogd, 2015 Family CLATHRINIDAE Minchin, 1900 Genus Clathrina Gray, 1867 sensu Klautau, Azevedo, Cóndor-Luján, Rapp, Collins & Russo, 2013 Clathrina fakaravae sp. nov. Clathrina huahineae sp. nov. Genus Ernstia Klautau, Azevedo, Cóndor-Luján, Rapp, Collins & Russo, 2013 Ernstia variabilis sp. nov. Family LEUCASCIDAE Dendy, 1892 Genus Leucascus Dendy, 1892 Leucascus digitiformis sp. nov. Leucascus simplex Dendy, 1892 Family LEUCETTIDAE De Laubenfels, 1936 Genus Leucetta Haeckel, 1872 Leucetta chagosensis Dendy, 1913 Leucetta microraphis Haeckel, 1872 Subclass CALCARONEA Bidder, 1898 Order LEUCOSOLENIDA Hartman, 1958 Family GRANTIIDAE Dendy, 1892 Genus Leucandra Haeckel, 1872 Leucandra tahuatae sp. nov.

Morphological taxonomy

Ascandra cf. crewsi Van Soest & De Voogd, 2015 (Fig 2; Table 2)

Synonym. Ascandra crewsi, Van Soest & De Voogd 2015: 36.

Material examined: UFRJPOR 6462 = MNHN-IP-2018-33—Tairineneva, Raiatea, Society Islands, station SR02 (16° 45.326’ S–151° 29.827’ W), depth: 15 m, coll. C. Debitus, 12/VIII/2009, P53. UFRJPOR 6463 = MNHN- IP-2018-34, UFRJPOR 6464 = MNHN-IP-2018-35—Tahiti, Society Islands, station ST22 (17° 32.485’ S–149° 35.205’ W), depth: 14 m, coll. C. Debitus, 29/V/2009, P23. UFRJPOR 8916 = MNHN-IP-2018-57—Tahiti, Soci- ety Islands, station ST27 (17°46.642’ S–149°24.236’ W), coll. S. Petek, 13/IV/2013, depth: 30 m, P485. UFRJPOR 8917 = MNHN-IP-2018-58—Tahiti, Society Islands, station ST29 (17°53.052’ S–149°11.411’ W), coll. S. Petek, 14/IV/2013, depth: 20 m, P507. UFRJPOR 8920 = MNHN-IP-2018-61—Tahiti, Society Islands, station ST59 (17°47.343’S–149°27.229’W), coll. C. Debitus, 13/IV/2013, depth: 15 m. Colour. White alive and white or light brown in ethanol (Fig 2A). Morphology and anatomy. Cormus delicate, formed by large, irregular, loosely anastomosed and ramified tubes with one or several oscula at the end of larger tubes (water-collecting tubes; Fig 2A). Aquiferous system asconoid. The skeleton is composed of two size categories of tetractines (Fig 2B), of which the larger is rare, and by small rare triactines. Spicules (Table 2) Tetractines I. Large. Regular (equiangular and equiradiate) or sagittal. Actines are conical with sharp tips (Fig 2C). The sagittal spicules sometimes have curved paired actines. The apical actine is cylindrical, very long, sharp, smooth and frequently curved but straight apical actines are also present (Fig 2C). Size: 162.0–218.7/ 16.2–27.0 µm (basal), 75.0–150.0/ 5.0–15.0 µm (apical). Tetractines II. Small. Regular (equiangular and equiradiate) or sagittal. They are very similar to Tetractines I, but a little smaller and with thinner actines (Fig 2 D). Size: 110.7–178.2/ 10.8–16.2 µm (basal), 17.5–234.9/ 1.3–9.5 µm (apical).

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 267 Triactines. Rare, small. Regular (equiangular and equiradiate) or sagittal. Actines are conical with sharp tips (Fig 2E). The sagittal spicules sometimes have curved paired actines. Size: 77.5–137.5/ 8.8–10.0 µm. Ecology. This sponge was found in holes or below dead corals, in a slightly muddy environment. Geographical distribution. Papua New Guinea (type locality; Van Soest & De Voogd 2015) and Society Is- lands (Raiatea and Tahiti—present work).

FIGURE 2. Ascandra cf. crewsi (UFRJPOR 8920). A—Specimen in vivo. B—Section. C—Tetractine I. D—Tetractines II. E—Triactines.

268 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. Remarks. Eleven species of Ascandra are currently known: Ascandra falcata Haeckel, 1872 from Lesina, Adriatic Sea (the type species of the genus); A. ascandroides (Borojević, 1971) from Rio de Janeiro, Brazil; A. atlantica (Thacker, 1908) from Cape Verde Islands; A. biscayae (Borojević & Boury-Esnault, 1987) from Bay of Biscay; A. brandtae (Rapp, Göcke, Tendal & Janussen, 2013) from the Weddell Sea, Antarctica; A. contorta (Bowerbank, 1866) from the Mediterranean Sea; A. corallicola (Rapp, 2006) Trondheimsfjord, Norway; A. crewsi Van Soest & De Voogd, 2015 from Wahoo, Papua New Guinea; A. densa Haeckel, 1872 from South Australia; A. kakaban Van Soest & De Voogd, 2015 from Kalimantan, Indonesia; and A. minchini Borojević, 1966 from the Mediterranean Sea. Considering cormus and skeleton composition (triactines and two size categories of tetractines), the specimens from the French Polynesia are more similar to A. crewsi and A. kakaban. The latter, however, has the same proportion of triactines and tetractines, while A. crewsi has very few triactines, like our specimens. Therefore, although the spicules of the specimens from French Polynesia are a little larger and have tetractines with actines more conical than those of A. crewsi [A. crewsi—Triactines: 140.0˗150.0/ 10.0˗12.0 µm; Tetractines I: 159.0˗206.4˗246.0/ 15.0˗18.8˗21.0 µm (basal actines), 181.0˗226.3˗279.0/ 13.0˗15.3˗17.0 µm (apical actine); Triactines II: 54.0˗90.2˗117.0/ 7.0˗7.3˗8.0 µm (basal actines), 62.0˗95.8˗114.0/ 2.5˗3.1˗3.5 µm (apical actine)], we think they can be conspecific.

TABLE 2. Spicule measurements of Ascandra cf. crewsi from French Polynesia. Spicule Actine Length (µm) Width (µm) N min mean sd max min mean sd max UFRJPOR 6463 Triactine Basal 77.5 112.0 20.1 137.5 8.8 9.8 0.5 10.0 11 Tetractine I Basal 162.0 180.8 9.9 205.2 18.9 21.4 2.2 24.3 20 Apical 75.0 112.0 23.5 150.0 7.5 12.1 2.2 15.0 10 Tetractine II Basal 110.7 139.7 14.0 159.3 10.8 14.0 1.3 16.2 20 Apical 116.1 160.5 33.3 234.9 8.1 8.3 0.5 9.5 11 UFRJPOR 8920 Triactine Basal 110.0 132.9 12.5 145.0 9.0 9.9 0.4 10.0 07 Tetractine I Basal 180.9 200.4 11.8 218.7 16.2 20.6 3.4 27.0 13 Apical 95.0 109.0 16.6 137.5 5.0 8.5 2.9 12.5 05 Tetractine II Basal 121.5 150.8 16.7 178.2 10.8 13.4 0.6 13.5 20 Apical 17.5 53.1 11.4 70.0 1.3 4.6 1.1 7.5 21

Clathrina fakaravae sp. nov. (Fig 3, Table 3)

Etymology. From the type locality (Fakarava) Type locality. Maruka (Fakarava). Tuheiava (Tikehau). Avatoru (Rangiroa) Tuamotu Islands. French Polynesia. Material examined. Holotype: UFRJPOR 6884 = MNHN-IP-2018-49—Rangiroa, Tuamotu Islands, station TRAN05 (14°56.481’ S–147°51.752’ W), depth: 24 m, coll. E. Folcher, 23/V/2011, P322. Paratype: UFRJPOR 6873 = MNHN-IP-2018-38—Fakarava, Tuamotu Islands, Station TFAK08 (16°08.230’ S–145°49.323’ W), depth: 15 m, coll. A. Renaud, 19/V/2011, P305˗TFAK08. Other material: UFRJPOR 6875 = MNHN-IP-2018-40—Tike- hau, Tuamotu Islands, station TTIK03 (14°59.629’ S–148°16.922’ W), depth: 40 m, coll. E. Folcher, 29/V/2011, P305˗TTIK03. Diagnosis. White Clathrina with large and loosely anastomosed tubes, no water-collecting tubes and three categories of triactines: conical, slightly conical and cylindrical. Colour. White alive and beige to light yellow in ethanol (Fig 3A). Morphology and anatomy. The cormus of this species is formed by large, irregular and loosely anastomosed tubes but in some regions the anastomosis is very tight, giving an appearance of a continuous membrane. Water- collecting tubes were not observed. Aquiferous system asconoid. The skeleton is composed of two categories of triactines, one cylindrical to slightly conical (the most abundant) and the other conical (Fig 3B).

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 269 Spicules (Table 3) Triactines I. Regular (equiangular and equiradiate). Actines are cylindrical to slightly conical with sharp tips (Fig 3E). Size: 95.7/ 7.2 µm. Triactines II. Regular (equiangular and equiradiate). These spicules are larger. Actines are conical with sharp tips (Fig 3D). Size: 159.8/ 12.3 µm. Geographical distribution. Rangiroa, Tuamotu Islands; Fakarava, Tuamotu Islands; Tikehau, Tuamotu Is- lands. Remarks. Although the tight anastomosis of the tubes in some parts of the cormus gives the impression of a begining of a continuous membrane formation, there is no cavity (pseudoatrium), therefore, it could not be an As- caltis. As it seems to us that there is no true continuous membrane, we consider we have a true Clathrina, perhaps with a beginning of cortex formation (cortical membrane). Unfortunately, for this species we were not able to get DNA sequence. Clathrina fakaravae sp. nov. has two categories of triactines (cylindrical to slightly conical and conical) and white cormus formed by irregular and loosely anastomosed tubes without water-collecting tubes, hence, we compared it with: C. rotundata Voigt et al., 2017 and C. zelinhae Azevedo et al., 2017. Clathrina rotundata, however, has also parasagittal triactines and actines with rounded tips, while the new species has always regular spicules with sharp tips. Clathrina zelinhae has tightly anastomosed tubes and the difference between the size of its conical and cylindrical spicules is much larger than in C. fakaravae sp. nov. [C. fakaravae sp. nov.—Triactines with cylindrical actines: 95.7 (±7.2)/ 7.2 (±1.5) µm; Triactines with conical actines: 159.8 (±13.2)/ 12.3 (±1.4) µm; C. zelinhae (holotype)—Triactines with cylindrical actines: 95.8 (±5.4)/ 3.9 (±1.0) µm; Triactines with conical actines: 271.1 (±13.2)/ 20.8 (±2.2) µm]. Therefore, our specimens constitute a new species for science.

FIGURE 3. Clathrina fakaravae sp. nov. (UFRJPOR 6884). A—Fixed specimen. B—Section. C—Triactines I. D—Triactine II.

270 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. TABLE 3. Spicule measurements of the holotype of C. fakaravae sp. nov. (UFRJPOR 6884). Spicule Length (µm) Width (µm) N min mean sd max min mean sd max Triactine I 43.2 95.7 7.2 135.0 5.4 7.2 1.5 10.8 30 Triactine II 137.7 159.8 13.2 180.9 10.8 12.3 1.4 13.5 20

Clathrina huahineae sp. nov. (Fig 4; Table 4)

Synonym. Clathrina sp. nov. 5, Klautau et al. 2013: 449.

Etymology. From the type locality (Huahine Island). Type locality. Fare. Huahine Island, Society Islands, French Polynesia. Material examined. Holotype: UFRJPOR 6461 = MNHN-IP-2018-32—Huahine Island, Society Islands, Sta- tion SH02 (16° 42.596’ S–151° 02.640’ W), depth: 14 m, coll. C. Debitus, 20/VIII/2009, P99. Teavonae (Takaroa). Avatoru (Rangiroa). Mehetia. French Polynesia. Paratype: UFRJPOR 6886 = MNHN-IP-2018-51—Mehetia, Society Islands, Station SME02 (17°53.110’ S–148°04.454’ W), depth: 16 m, coll. E. Folcher, 26/IV/2011, P228– SME02. Other material: UFRJPOR 6879 = MNHN-IP-2018-44—Takaroa, Tuamotu Islands, Station TTAK01 (14°27.717’ S–145°02.356’ W), depth: 12 m, coll. D. Fleurisson, 14/V/2011, P228-TTAK01. UFRJPOR 6880 = MNHN-IP-2018-45—Rangiroa, Tuamotu Islands, Station TRAN07 (14°56.215’ S–147°42.024’ W), depth: 30 m, coll. A. Renaud, 24/V/2011, P228-TRAN07. UFRJPOR 8954 = MNHN-IP-2018-63—raroia, tuamotu Islands, Station trar15 (16°09.586’ S 142°31.998’ W), depth: 15 m, coll. M. Dumas, 07/XI/2018, P667-trar15. UFR- JPOR 8957 = MNHN-IP-2018-66—Makemo, tuamotu Islands, Station tMaK09 (16°38.948’ S–143°33.333’ W), depth: 23 m, coll. S. Petek, 08/XI/2018, P671-tMaK09. Diagnosis. Yellow Clathrina with large and loosely anastomosed tubes, water-collecting tubes and triactines with very thin, cylindrical actines and rounded to blunt tips. Colour. Yellow alive and beige in ethanol (Fig 4A). Morphology and anatomy. The cormus of this species is massive but delicate, formed by large, irregular, and loosely anastomosed tubes. Large water-collecting tubes are present. Aquiferous system asconoid. The skeleton is composed of two size categories of triactines (Fig B). The triactines I are smaller (Fig 4C) and the triactines II are larger and more abundant (Fig 4D). Spicules (Table 4) Triactines I. Regular (equiangular and equiradiate). Actines are conical, with sharp tips (Fig 4C). Size: 65/ 7.1 µm. Triactines II. Regular (equiangular and equiradiate), subregular or parasagittal. Actines are cylindrical, very thin, with rounded to blunt tips (Fig 4D). The spicules with rounded tips seem to be larger. Frequently actines are undulated. Size: 142.4/ 7.5 µm. Geographical distribution. Huahine Island, Society Islands; Teavonae (Takaroa); Avatoru (Rangiroa); Mehe- tia; Takaroa, Tuamotu Islands; Rangiroa, Tuamotu Islands. Remarks. According to our molecular analysis, Clathrina huahineae sp. nov. is sister species of the Tropical Western Atlantic species C. mutabilis Azevedo et al., 2017. They are morphologically very similar. Both are yellow and formed by large, loose and irregularly anastomosed tubes and present water-collecting tubes. Besides, they have two kinds of spicules, a regular small triactine with conical actines and sharp tips, which is less abundant, and an abundant regular, subregular or parasagittal large triactine with cylindrical actines. Their only morphological differences are the post-fixation colour, which is white in C. mutabilis and beige in C. huahineae sp. nov., and the tip of the triactines II, which are blunt to rounded in the new species and blunt to sharp in C. mutabilis. We also found a slight difference in the size of the spicules, thicker in C. mutabilis (Holotype, UFRJPOR 6526—Triactine I: 56.7–69.8–91.8/ 8.1–8.4–9.5 µm; Triactine II: 94.5–121.7–148.5/ 6.8–8.1–9.5 µm). The other species of Clathrina morphologically similar to the new species are C. beckingae Van Soest & De Voogd, 2015 and C. purpurea Van Soest & De Voogd, 2015. Unfortunately, they could not be molecularly compared because there are no ITS sequences available of those species. Both C. beckingae and C. purpurea have cormus

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 271 formed by irregular and loosely anastomosed tubes, water-collecting tubes and triactines with very thin cylindrical actines (6 µm). Those species are from Indonesia and can be differentiated from C. huahineae sp. nov. by the colour, which is yellow in our species, white in C. beckingae and reddish purple in C. purpurea (Van Soest & De Voogd 2015). Besides, C. purpurea does not have water-collecting tubes and C. beckingae has smaller spicules (48.0–84.9–106.0/ 6.0 µm).

FIGURE 4. Clathrina huahineae sp. nov. (UFRJPOR 6461). A—Fixed specimen. B—Section. C—Triactine I. D—Triactines II.

TABLE 4. Spicule measurements of Clathrina huahineae sp. nov. H = holotype. P = paratype. Spicule Length (µm) Width (µm) N min mean sd max min mean sd max UFRJPOR 6461 (H) Triactine I 40.0 65.0 16.3 90.0 5.0 7.1 0.8 7.5 20 Triactine II 110.0 142.4 14.9 175.0 7.5 7.5 0.9 7.5 20 UFRJPOR 6886 (P) Triactine I 25.0 69.9 18.9 95.0 5.0 7.4 0.9 10.0 20 Triactine II 120.0 141.9 12.5 165.0 6.3 7.4 0.5 8.8 20

Etymology. From the Latin “variabilis” (varied, changeable), due to its variable spicule shape. Type locality. Rairoa, Tuamotu Islands, French Polynesia Material examined. Holotype: UFRJPOR 8963 = MNHN-IP-2018-56—raroia, tuamotu Islands, Station trar08 (16°02.831’ S 142°25.701’ W), depth: 17 m, coll. M. Dumas, 05/XI/2018, P455-trar08. Paratype: UFR- JPOR 8918 = MNHN-IP-2018-59—Raivavae, Australes Islands, Station ARAI07 (23°53.282’ S–147°40.902’ W), depth: 6 m, coll. C. Debitus, 23/III/2013, P455. Other material. UFRJPOR 8962 = MNHN-IP-2018-71—raroia, tuamotu Islands, Station trar02 (16°01.99’ S 142°25.565’ W), depth: 20 m, coll. CS. Petek, 04/XI/2018, P455-trar02. UFRJPOR 8964 = MNHN-IP-2018-42—raroia, tuamotu Islands, Station trar09 (16°02.074’ S 142°25.701’ W), depth: 15 m, coll. V. Bouvot, 05/XI/2018, P455-trar09. UFRJPOR 8965 = MNHN-IP-2018- 52—raroia, tuamotu Islands, Station trar12 (16°00.896’ S 142°25.323’ W), depth: 20 m, coll. M. Dumas, 06/ XI/2018, P455-trar12. Diagnosis. Yellow Ernstia with spherical cormus and thin, regular and tightly anastomosed tubes. A single long osculum atop of and atrial cavity. Skeleton composed of two categories of triactines and tetractines, mainly differentiated by shape and size. Aquiferous system solenoid. Colour. Yellow alive and light brown in ethanol (Fig 5A). Morphology and anatomy. Cormus spherical to subspherical formed by thin, regular and tightly anastomosed tubes (Figs 5A, B). There is usually one long apical osculum (Fig 5C), with a continuous endopinacoderm lining the atrial cavity. Aquiferous system solenoid. The skeleton is composed of two categories of triactines, two of tetractines, and trichoxeas (Fig 5D). The apical actines of the tetractines project into the tubes lumen and atrial cavity. The size of triactines II and tetractines II is different depending on the regions of the sponge: the spicules in the oscular region are noticeably larger than the spicules in the other parts of the body. Due to its transitional changes from osculum to choanosome, we consider it the same variable category. Tetractines occur in higher frequencies than triactines. Spicules (Table 5) Triactines I. Regular (equiangular and equiradiate) and smaller than triactines II. This category ressembles a young spicule on formation, but due to its relatively abundance, we considered it as a category apart. Actines are conical with sharp tips (Fig 5E). Size: 41.4/ 5.3 µm. Triactines II. Regular (equiangular and equiradiate) and very abundant. Actines vary highly in shape and size. Spicules from the oscular region are usually cylindrical, ondulated and larger, with blunt tips. Spicules from choanosome are slightly conical to conical and straight, with blunt to slightly sharp tips (Fig 5E). Size: 94.3/ 6.9 µm. Tetractines I. Regular (equiangular and equiradiate) and smaller than tetractines II. This category ressembles a young spicule on formation, but due to its relatively abundance, we considered it as a category apart. Actines are conical with sharp tips. The apical actine is cylindrical, the same size of basal actines, sharp, smooth, and frequently curved (Fig 5G). Size: 43.0/ 5.1 µm (basal actine); 39.1/ 4.1 µm (apical actine). Tetractines II. Regular (equiangular and equiradiate) and very abundant. Actines vary highly in shape and size. Spicules from the oscular region are usually cylindrical, ondulated and larger, with blunt tips. Spicules from choanosome are slightly conical to conical and straight, with blunt to slightly sharp tips (Fig 5F). The apical actine is cylindrical, very long, sharp, smooth, and frequently curved (Fig 5G). Size: 90.6 / 7.0 µm (basal actine); 51.9/ 4.7 µm (apical actine). Geographical distribution. Raroia, Tuamotu Islands and Raivavae, Australes Islands (present work). Remarks. Ernstia variabilis sp. nov. formed a very well supported clade (100% bootstrap) with E. pyrum Sana- myan et al., 2019 and E. citrea Azevedo et al., 2017. The three species are morphologically almost identical, all of them having a spherical yellow cormus formed by tight and regularly anastomosed tubes, long apical osculum and solenoid aquiferous system. Although in the original description of E. citrea it was said that this species has asconoid aquiferous system, we re-analised it and found the membrane of pinacocytes surrounding the atrium, proving that its aquiferous system is solenoid, as Sanamyan et al. (2019) had observed for E. pyrum. We are describing E. variabilis sp. nov. as possessing two categories of triactine and two of tetractines. Re-analysing E. citrea, we think it has also these categories, though in the original description it was considered as having only one category of tri- and one of tetractines. The same can be considered for E. pyrum, if we take into account the size variation of the spicules and look at the original pictures (Sanamyan et al. 2019). Therefore, these three species can be distinguished only by slight differences in the size of their spicules.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 273 FIGURE 5. Ernstia variabilis sp. nov. (UFRJPOR 8963). A—Specimen in situ (UFRJPOR 8918). B—Fragment fixed in ethanol (arrow pointing to the osculum). C—Skeleton of the osculum. D—Skeleton of the anastomosed tubes. E—Triactine I. F—Triactine II. G—Tetractine I. H—Tetractine II. I—Apical actine of a tetractine.

Ernstia variabilis sp. nov. has spicules thicker than those of E. citrea [Holotype—Triactine: 81.3 (4.3)/ 10.4 (0.7), Tetractine: 82.4 (6.2)/ 10.4 (1.1)] (to compare with Table 5). Ernstia pyrum has a little thicker spicules [Ho- lotype—Triactine—surface and choanosome: 74.8 (14.4)/ 6.9, Triactine—atrial membrane: 83.7 (22.1)/ 7.9, Tri-

274 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. actine—osculum: 103.9 (30.5)/ 6.9 (1.0), Tetractine—surface and choanosome: 73.6 (13.7)/ 6.6, Tetractine—atrial membrane: 94.1 (24.5)/ 7.4)] (to compare with Table 5). Hence, the three species are almost cryptic, however, we decided to distinguish them based on the molecular tree (Fig 13). In the tree, although the three species form a very well supported clade (100% bootstrap), E. citrea and E. variabilis sp. nov. form two well supported clades (99% and 100% bootstrap, respectively). As E. variabilis sp. nov. is the third species of the genus with solenoid aquiferous system, we proposed an emendation to the current diagnosis: Calcinea in which the cormus comprises a typical clathroid body. A stalk may be present. The skeleton contains regular (equiangular and equiradiate) and/or sagittal triactines and tetractines. Tetractines are the most abundant spicules or occur at least in the same proportion as the triactines. Tetractines frequently have very thin (needle-like) apical actines. Diactines may be added. Asconoid or solenoid aquiferous system (Klautau et al. 2013).

TABLE 5. Spicule measurements of the holotype of Ernstia variabilis sp. nov. (UFRJPOR 8962). Length (µm) Width (µm) Spicule Actine min mean sd max min mean sd max N Triactine I Basal 30.0 41.4 7.8 62.5 3.8 5.3 0.9 7.5 20 Triactine II Basal 75.0 94.3 10.3 112.5 5.0 6.9 0.9 7.5 20 Tetractine I Basal 30.0 43.0 6.7 55.0 3.8 5.1 0.8 7.5 20 Apical 27.5 39.1 7.9 47.5 2.5 4.1 0.9 5.0 20 Tetractine II Basal 70.0 90.6 12.1 117.5 5.0 7.0 0.9 7.5 20 Apical 32.5 51.9 14.1 95.0 3.8 4.7 0.6 5.0 08

Leucascus digitiformis sp. nov. (Fig 6, Table 6)

Etymology. From the Latim “digitus” (finger), for the finger-shaped cormus of this species. Type locality. Tekeho (Nuku Hiva), Marquesas Islands. French Polynesia. Material examined. Holotype: UFRJPOR 6460 = MNHN-IP-2018-31—Nuku Hiva, Marquesas Islands, Sta- tion MNH04 (08° 57.661’ S–140° 10.149’ W), depth: 16 m, coll. C. Debitus, 30/VIII/2009, P114. Diagnosis. White Leucascus with digitiform cormus and large oscula. Skeleton composed of tripods, triactines and tetractines with large spines on the apical actine. Colour. White alive and in ethanol (Fig 6A). Morphology and anatomy. Sponge digitiform, massive but delicate (Figs 6A, B). The cormus is formed by thin, regular and tightly anastomosed tubes forming a continuous delicate cortex (Fig 6C). Each protuberance (digitus) present a terminal osculum surrounded by membrane. Below each osculum there is an atrial cavity supported by tetractines. Aquiferous system solenoid. The specimen is full of embryos. The skeleton is composed of tripods and triactines on the cortex (Fig 6C), triactines and tetractines in the choanosome (Fig 6D), and tetractines in the atrial wall. Spicules (Table 6) Tripods. Regular or sagittal. Similar to large triactines. Actines are conical with blunt tips (Fig 6E). Size: 88.0/ 9.3 µm. Triactines. Regular or sagittal. Actines are conical with blunt tips (Fig 6F). Size: 54.7/ 5.3 µm. Tetractines. Regular or sagittal. Actines are conical with blunt tips (Fig 6G). The apical actine is very long, thick, conical and sharp, covered by spines (Fig 6H). Size: 57.4/ 5.5 µm (basal actine); 35.4/ 5.0 µm (apical actine). Geographical distribution. Nuku Hiva, Marquesas Islands (present work). Remarks. Currently 10 species of Leucascus are recognised: L. simplex Dendy, 1892 (type species of the genus) from South Australia; L. albus Cavalcanti, Rapp & Klautau, 2013 from southeastern Brazil; L. clavatus Dendy, 1892 from South Australia; L. flavus Cavalcanti, Rapp & Klautau, 2013 from Indonesia; L. leptoraphis (Jenkin, 1908) from Antarctica; L. lobatus Rapp, 2004 from Greenland; L. neocaledonicus Borojević & Klautau, 2000 from New Caledonia; L. protogenes (Haeckel, 1872) from South Australia; L. roseus Lanna, Rossi, Cavalcanti, Hajdu & Klautau, 2007 from southeastern Brazil; and L. schleyeri Van Soest & De Voogd, 2018 from South Africa.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 275 FIGURE 6. Leucascus digitiformis sp. nov. (UFRJPOR 6460). A—Specimen in situ. B—Fixed specimen. C—Tangential section of the cortex. D—Cross-section of the choanosome. E—Tripods. F—Triactines. G—Tetractines. H—Apical actine of tetractines.

Considering skeleton composition, L. digitiformis sp. nov. is similar to L. leptoraphis and L. lobatus, as all of them have tripods, triactines and tetractines. The new species, however, differs from them by several characteris- tics. For example, L. leptoraphis and L. lobatus have triactines and tetractines with cylindrical actines, while our

276 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. new species has spicules with conical actines. Besides, in L. leptoraphis tetractines are very rare and in L. lobatus the tripods have a kind of rudimentary fourth actine in the tripods. Moreover, the spines of the apical actine of the tetractines of L. digitiformis sp. nov. differ from those of all the other species of Leucascus, as they are large in the new species. These large spines resemble those of Borojevia, however, the new species clearly has an atrial membrane, a characteristic of Leucascus and absent in Borojevia. Nonetheless, looking at Borojevia spp., we found that the habitus of Borojevia tubulata Van Soest & De Voogd, 2018, a species from Maldives, is very similar to ours. Besides, a picture showing the atrium of the specimen ZMA Por. 12435, from the Seychelles, suggests that that specimen has a true atrium, although this specimen had grouped molecularly inside Borojevia (Van Soest & De Voogd, 2018). We unfortunately did not succeed to get a DNA sequence of our specimen to compare it with B. tubulata. Therefore, we compared both species only morphologically and we found differences in the size of the spicules [B. tubulata—holotype—Tripods: 92˗133˗189/ 11˗13.4˗16; Triactines: 54˗63˗111/ 5.0˗6.3˗7.5; Tetractines: 51˗68˗96/ 6.0˗6.6˗9.0 (basal), 24˗37˗48/ 3.5˗4.4˗5.0 (apical)] (Table 6). As we found these differences in the size of the spicules and as Van Soest & De Voogd (2018) stated that their species do not have a true atrium, we decided to consider our species as a new one, still, it is desirable in the future to compare them molecularly.

TABLE 6. Spicule measurements of the holotype of Leucascus digitiformis sp. nov. (UFRJPOR 6460). Spicule Actine Length (µm) Width (µm) N min mean sd max min mean sd max Tripod Basal 59.4 88.0 11.6 108.0 8.1 9.3 1.4 10.8 20 Triactine Basal 45.9 54.7 4.5 64.8 2.7 5.3 0.6 5.4 20 Tetractine Basal 51.3 57.4 3.5 67.5 5.4 5.5 0.6 8.1 20 Apical 13.5 35.4 18.4 75.6 2.7 5.0 1.6 8.1 10

Leucascus simplex Dendy, 1892 (Fig 7; Table 7)

Synonyms. Leucascus simplex, Dendy 1892: 77, Kirk 1897: 313, Dendy 1913: 9, Dendy & Row 1913: 731, Row & Hôzawa 1931: 742, Cavalcanti et al. 2013: 277. Leucetta chagosensis, Hall et al. 2013: 500.

Type locality. Port Phillip Heads, Australia Material examined. UFRJPOR 6451 = MNHN-IP-2018-22—Tahiti, Society Islands, Station ST12 (17°31.30’ S–149°33.40’W), depth: 10 m, coll. C. Debitus, 23/III/2009, P2. UFRJPOR 6456 = MNHN-IP-2018-27—Hiva Oa, Marquesas Islands, Station MHO05 (9° 42.553’ S–139° 01.18’ W), depth: 15 m, coll. C. Debitus, 08/IX/2009, P167. Other material. UFRJPOR 6458 = MNHN-IP-2018-29—Moorea, Society Islands, Station M01 (17°29.681’ S– 149°51.717’ W), depth: 8 m, coll. C. Debitus, 04/XII/2010, P221. UFRJPOR 8919 = MNHN-IP-2018-60—Tahiti, Society Islands, Station ST52 (17°47.147’ S–149°25.359’ W), depth: 30 m, coll. S. Petek, 21/IV/2013, P480˗ST52. UFRJPOR 8921 = MNHN-IP-2018-62—Tahiti, Society Islands, Station ST27 (17°46.642’ S–149°24.236’ W), depth: 20 m, coll. S. Petek, 12/IV/2013, P480˗ST27. Colour. White alive and beige to light yellow in ethanol (Fig 7A). Morphology and anatomy. Sponge massive, almost spherical. Sometimes there is more than one “sphere” attached to another (Figs 7A, B). Each sphere has an apical osculum sometimes surrounded by membrane. The cormus is formed by regular and tightly anastomosed tubes covered by a continuous, smooth, delicate cortex. The tubes near the surface are more parallel to each other than in the middle of the choanosome. The atrial cavity is large and delimited by endopinacoderm. Aquiferous system solenoid. The skeleton is composed of triactines and tetractines (Fig 7C). Triactines are more abundant than the tetractines. The atrial skeleton is composed of tetractines only. Spicules (Table 7) Triactines. Regular to sagittal. Actines are slightly conical with sharp tips (Fig 7D). Size: 102.5/ 8.7 µm. Tetractines. Regular to sagittal. Actines are slightly conical with sharp tips (Fig 7E). The apical actine is very thin (needle-like), cylindrical and sharp, covered with very short spines (Fig 7F). Size: 96.7/ 8.1 µm (basal actine); 72.8/ 2.7 µm (apical actine).

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 277 FIGURE 7. Leucascus simplex (UFRJPOR 6451). A—Specimen in situ. B—Fixed specimen. C—Cross-section of the choanosome. D—Triactine. E—Tetractine. F—Apical actine of two tetractines. Abbreviation: at = atrium; cx = cortex.

Geographical distribution. Tahiti and Moorea, Society Islands; Hiva Oa, Marquesas Islands. XPirae (Tahiti- ST12). Mataeia (Rautirae. Tahiti-ST27). Mataeia (Aifa. Tahiti-ST52). Opunohu (Moorea). Hanamenu (Hiva Oa). French Polynesia and Marquesas Islands (Hall et al. 2013). Remarks. The specimens from French Polynesia have spicules with sharp tips, while in the holotype they are blunt. Besides, in the holotype the tetractines are rare, while in the specimens from French Polynesia they are abun-

278 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. dant (although still less abundant than the triactines). The spicules from the french polynesian specimens are also thinner than those of the holotype [Triactines: 83.2–102.9 (±6.7)–114.4/ 11.7–13.3 (±0.9)–15.9 µm; Tetractines: 72.8–100.1 (±9.2)–117.0/ 10.4–12.1 (±1.0)–14.3 µm (basal), 26.0–42.3 (±8.8)–65.0/ 2.6–4.0 (±1.0)–5.2 µm (apical)] (Table 7). In our C-LSU molecular tree L. simplex is sister species of L. flavus.

TABLE 7. Spicule measurements of Leucascus simplex (UFRJPOR 6451). Spicule Actine Length (µm) Width (µm) N min mean sd max min mean sd max Triactine Basal 81.0 102.5 14.8 148.5 8.1 8.7 1.2 10.8 21 Tetractine Basal 27.0 96.7 17.0 108.0 5.4 8.1 0.9 10.8 21 Apical 51.3 72.8 14.1 108.0 2.7 2.7 0.0 2.7 20

Leucetta chagosensis Dendy, 1913 (Fig 8, Table 8)

Synonyms. Leucetta chagosensis—Dendy 1913: 10, Dendy & Row 1913: 733, Dendy & Frederick 1924: 482, Burton 1963: 241, Borojević 1967: 2, Pulitzer-Finali 1982: 89, Gosliner et al. 1996: 16, Lévi et al. 1998: 77, Wörheide & Hooper 1999: 882, Borojević & Klautau 2000: 194, Wörheide et al. 2002: 1753, Wörheide et al. 2005: 379, Baine & Harasti, 2007: 15, Wörheide et al. 2008: 1, Voigt et al. 2012a: 101, Van Soest & De Voogd 2015: 51, 2018: 76; L. infrequens—Row & Hôzawa 1931: 747, Burton 1963: 241, Borojević & Klautau 2000: 195; L. expansa—Row & Hôzawa 1931: 749, Burton 1963: 241; Ascoleucetta sagittata Cavalcanti et al. 2013: 308, Van Soest & De Voogd 2015: 49; Leucetta sp.—Colin & Arneson 1995: 60 (photo 230).

Material examined. UFRJPOR 6455 = MNHN-IP-2018-26—Moorea, Society Island, Station SM01 (17°29.681’ S–149°51.717’ W), depth: 10 m, coll. C. Debitus, 04/XII/2010, P2- SM01. UFRJPOR 6889 = MNHN-IP-2018- 54—Makemo, Tuamotu Islands, Station TMAK06 (16°28.120’ S–143°57.200’ W), depth: 18 m, coll. B. Bour- geois, 10/V/2011, P266. UFRJPOR 6871 = MNHN-IP-2018-36—Fakarava, Tuamotu Islands, Station TFAK04 (16°05.231’ S–145°44.127’ W), depth: 50 m, coll. B. Bourgois, 18/V/2011. UFRJPOR 6874 = MNHN-IP-2018- 39—Rangiroa, Tuamotu Islands, Station TRAN04 (15°05.314’ S–147°56.531’W), depth: 30 m, coll. E. Folcher, 23/V/2011. UFRJPOR 6876 = MNHN-IP-2018-41—Fakarava, Tuamotu Islands, Station TFAK02 (16°04.900’ S–145°41.497’ W), depth: 15m, coll. S. Petek, 17/V/2011. UFRJPOR 6878 = MNHN-IP-2018-43—Rangiroa, Tuamotu Islands, Station TRAN01 (15°13.359° S–147°14.832’ W), depth: 40 m, coll. B. Bourgeois, 22/V/2011. UFRJPOR 6882 = MNHN-IP-2018-47—Toau, Tuamotu Islands, Station TTOA03 (15°47.480’ S–145°55.120’ W), depth: 18 m, coll. A. Renaud, 21/V/2011. UFRJPOR 6885 = MNHN-IP-2018-50—Tetiaroa, Society Islands, Station STET01 (17°02.258 S–149°33.707 W), depth: 40 m, coll. A. Renaud, 31/V/2011, P266˗STET01. UFR- JPOR 8955 = MNHN-IP-2018-64—Makemo, Tuamotu Islands, Station TMAK11 (16°38.485’ S–143°387.934’ W), depth: 30 m, coll. M. Dumas, 08/XI/2018, P669-TMAK11. UFRJPOR 8956 = MNHN-IP-2018-65—Makemo, Tuamotu Islands, Station TMAK11 (16°38.485’ S–143°38.934’ W), depth: 20 m, coll. S. Petek, 08/XI/2018, P670- TMAK11. UFRJPOR 8958 = MNHN-IP-2018-67—Rangiroa, Tuamotu Islands, Station TRAN06 (14°55.927° S–147°43.329’ W), depth: 30 m coll. M. Dumas, 13/XI/2018, P669-TRAN06. UFRJPOR 8959 = MNHN-IP- 2018-68—Rangiroa, Tuamotu Islands, Station TRAN18 (14°55.841° S–147°44.019’ W), depth: 15 m, coll. M. Dumas, 11/XI/2018, P669-TRAN18. UFRJPOR 8960 = MNHN-IP-2018-69—Rangiroa, Tuamotu Islands, Station TRAN18 (14°55.841° S–147°44.019’ W), depth: 15 m, coll. M. Dumas, 11/XI/2018, P670-TRAN18. Comparative material. BMNH 1920.12.9.51 (holotype). Colour. Yellow alive, beige in ethanol (Fig 8A). Morphology and anatomy. Sponge massive, spherical to sub-spherical, covered by a smooth cortex. Apical osculum surrounded by membrane (Fig 8A). Hard and friable. There are subcortical inhalant cavities and the canals are disposed in parallel, giving a radial organisation to the choanosome. Large exhalant canals arrive into the atrium, giving it a reticulated appearance. Aquiferous system leuconoid. The skeleton is composed of giant triactines, present mainly on the cortex, and small triactines and tetractines (Fig 8B). The small triactines are present on the cortex and choanosome, while the small tetractines, which are few, are found only in the choanosome, mainly in the exhalant canals.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 279 Spicules (Table 8) Giant triactines. Regular. Actines are conical with sharp tips (Fig 8E). Variable sizes. Size: 523.0/ 40.0 µm. Triactines. Regular to subregular. Actines are conical with sharp tips but some few spicules have cylindrical actines. The apical actine of the tetractines is conical and sharp (Fig 8F, I). Size: 139.1/ 13.6 µm.

FIGURE 8. Leucetta chagosensis (UFRJPOR 6455). A—Fixed specimen. B—Cross-section of the choanosome. The asterisk indicates the atrial skeleton. C—Tangential section of the cortex. D—Tangential section of the atrium. E—Giant triactine. F—Small triactines. G—Small tetractine. H—Apical actine of a tetractine. Abbreviation: cx = cortex.

280 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. Tetractines. Regular to subregular. They are very few. Actines are conical with sharp tips but some have cylindrical actines. The apical actine is conical, sharp, smooth and curve (Fig 8G). Size: 107.8/ 10.0 µm (basal actine); 32.0/ 5.2 µm (apical actine; Fig 8H). Geographical distribution. Indian Ocean—Chagos Archipelago (Dendy 1913), Abrolhos Islands (Dendy & Frederick 1924), Red Sea and Maldives (Wörheide et al. 2008, Voigt et al. 2012a); Western Pacific Ocean—Indo- nesia, Philippines, Japan, Western Australia, French Polynesia, and Samoa (Wörheide & Hooper 1999, Wörheide et al. 2002, 2005, 2008, Voigt et al. 2012a), New Caledonia (Borojević 1967, Lévi et al. 1998, Borojević & Klautau 2000). Remarks. Leucetta chagosensis is a species originally described from Chagos Archipelago (Indian Ocean). Although in the original description Dendy (1913) had mentioned only the presence of large and small triactines, we found few tetractines in the holotype (Table 8) and these spicules are being observed in all specimens identified as L. chagosensis. This species is considered to be widespread in the Indo-Pacific where it is characterised by its bright yellow colour when alive, lobose habitus, large triactines and small triactines and tetractines, the latter being rare. It is possible that L. chagosensis represents a species complex (Wörheide et al. 2002, 2005, 2008; Voigt et al. 2012a), however, as this putative species complex has not been solved yet, we call our specimens L. chagosensis.

Table 8. Spicule measurements of Leucetta chagosensis (UFRJPOR 6455). Spicule Actine Length (µm) Width (µm) N min mean sd max min mean sd max Giant triactine Basal 350.0 523.0 115.4 710.0 30.0 40.0 13.3 70.0 10 Triactine Basal 77.5 139.1 22.6 177.5 7.5 13.6 2.5 17.5 23 Tetractine Basal 65.5 107.8 21.4 155.0 5.0 10.0 2.3 15.0 30 Apical 17.5 32.0 10.5 50.0 5.0 5.2 0.8 7.5 11

Leucetta microraphis Haeckel, 1872 (Fig 9, Table 9)

Synonyms. Dyssycus primigenius, Lipostomella primigenia, Amphoriscus primigenius, Coenostomus primigenius, Artynas primigenius, Aphroceras primigenium, Leucometra primigenia—Haeckel 1872: 118; Leucetta primigenia var. microraphis—Haeckel 1872: 118, Ridley 1884: 482; Leucetta microraphis—Von Lendenfeld 1885: 1117, Dendy & Row 1913: 734, Dendy & Frederick 1924: 482, Row & Hôzawa 1931: 746, Tanita 1942: 111, Burton 1963: 270, Borojević 1967: 3, Borojević & Peixinho 1976: 1003, Pulitzer-Finali 1982: 87 (Pericharax orientalis according to Wörheide & Hooper 1999), Borojević & Klautau, 2000: 193, Wörheide & Hooper 1999: 879, Van Soest & De Voogd 2015: 54, 2018: 80; Leucan- dra primigenia var. microraphis—Row 1909: 186; Leucandra microraphis—Dendy 1892: 104; Leuconia dura—Poléjaeff 1883: 65, Lendenfeld 1885: 1118, Dendy 1892: 104; Leucaltis floridana var. australiensis—Carter 1886: 145; Leucandra carteri Dendy 1892: 103; Leucetta carteri—Dendy & Row 1913: 734, Burton 1963: 241, Borojević 1967: 5; Leucetta primigenia—Colin & Arneson 1995: 60 (photo 229); Gosliner et al. 1996: 16 (photo 4), Erhardt & Baensch 1998: 22 apud Van Soest & De Voogd 2015. The same authors mentioned that the specimens identified by Breitfuss (1896, 1898) as Leucetta solida were probably L. microraphis.

Material examined. UFRJPOR 6450 = MNHN-IP-2018-21—Bora Bora, Society Island (Leeward), ST SBB1 (16° 28.773’ S–151° 41.287’ W), depth: 15 m, coll. J. Orempuller, 09/VIII/2009, P29. UFRJPOR 6452 = MNHN- IP-2018-23˗ Tahiti Island (17° 32.140’ S–149° 35.449’ W), depth: 25 m, coll. C. Debitus, 28/V/2009, P19, ST21. UFRJPOR 6453 = MNHN-IP-2018-24—Tahiti, Society Islands, ST2 (17° 31.225’ S–149° 33.220’ W), depth: 14 m, coll. C. Debitus, 24/III/2009, P8˗ST2. UFRJPOR 6457 = MNHN-IP-2018-28—Moorea, Society Islands, Station SM01 (17°29.681’ S–149°51.717’ W), depth: 14 m, C. Debitus, 04/XII/2010, P8˗SM01. UFRJPOR 6459 = MNHN-IP-2018-30—Raiatea, Society Island, Station SR10 (16° 49.873’ S–151° 20.825’ W), depth: 40 m, coll. C. Debitus, 16/VIII/2010, P64. UFRJPOR 6881 = MNHN-IP-2018-46—Tetiaroa, Society Islands, Sta- tion STET01 (17°02.258 S–149°33.707 W), depth: 35 m, coll. E. Folcher, 31/V/2011, P348˗STET01. UFRJPOR 6888 = MNHN-IP-2018-53—Tetiaroa, Society Islands, Station STET03 (16°58.916 S–149°34.559 W), depth: 40 m, coll. D. Fleurisson, 01/VI/2011, P348˗STET03. UFRJPOR 8961—tetiaroa, Society Islands, Station ttet02 (16°59.967S–149°35.440W), depth: 28 m, coll. M. Dumas, 21/XI/2018, P678˗TTET02.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 281 Colour. White to light blue or violet alive (Fig 9A) and beige to light brown in ethanol (Fig 9B). Morphology and anatomy. This species is massive, hard and friable. It is harsh to the touch and has an amor- phous shape (Figs 9A, B). It has several apical oscula, surrounded by membrane, and large atrium. The canals are visible through the cortex, giving an anastomosed appearance to the sponge. Aquiferous system leuconoid (Fig 9C). The specimen UFRJPOR 6450 has embryos.

FIGURE 9. Leucetta microraphis (UFRJPOR 6450). A—Specimen in situ. B—Fixed specimen. C—Cross-section of the choanosome. D—Tangential section of the cortex. E—Tangential section of the atrium. F—Large triactines. G—Small triactines. H—Small tetractines. I—Apical actine of a tetractine. Abbreviation: at = atrium; cx = cortex.

282 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. The skeleton is composed of giant triactines present in the cortex (Fig 9D) and choanosome and small triactines and tetractines (Fig 9E). The small triactines are present in the cortex and choanosome, while the small tetractines are found only in the choanosome. The tetractines are very few and they are present only surrounding the canals. Spicules (Table 9) Giant triactines. Regular (equiangular and equiradiate) and subregular (not equiradiate). Actines are conical with sharp tips (Fig 9F). Variable sizes. Size: 1108.3/ 137.5 µm. Triactines. Regular to sagittal. Actines are conical with blunt tips (Fig 9G) but some triactines with cylindrical actines were also observed (Fig 9I). Size: 145.4/ 15.0 µm. Tetractines. Regular to sagittal. Actines are conical with blunt tips (Fig 9H) but some tetractines have also cylindrical actines (Fig 9J). The apical actine is very thin, conical, sharp, and smooth (Fig 9K). Sometimes it is undulated Size: 136.3/ 10.4 µm (basal actine); 30.8/ 5.9 µm (apical actine). Geographical distribution. Indo-Pacific (Poléjaeff 1883, Ridley 1884, Von Lendenfeld 1885, Carter 1886, Dendy 1892, Row 1909, Dendy & Frederick 1924, Row & Hôzawa 1931, Colin & Arneson 1995, Gosliner et al. 1996, Erhardt & Baensch 1998, Wörheide & Hooper 1999, Borojević & Klautau 2000, Van Soest & De Voogd 2015, 2018). Remarks. Leucetta microraphis is a widespread species with conserved skeleton but variable habitus. As these variable habitus have been considered only polymorphism (except for L. sulcata Van Soest & De Voogd, 2018, see Discussion), we identified our specimens as L. microraphis, however, it is possible that we have a new species of Leucetta in French Polynesia. We discuss this subject in more detail at the end of the paper.

TABLE 9. Spicule measurements of Leucetta microraphis (UFRJPOR 6453). Spicule Actine Length (µm) Width (µm) N min mean sd max min mean sd max Giant triactine Basal 925.0 1108.3 179.4 1400.0 125.0 137.5 20.9 175.0 06 Triactine Basal 114.8 145.4 21.4 232.2 7.5 15.0 11.6 21.6 42 Tetractine Basal 83.7 136.3 10.4 162.0 5.4 10.4 1.4 12.5 42 Apical 10.8 30.8 17.2 67.5 4.1 5.9 1.9 10.8 17

Leucandra tahuatae sp. nov. (Figs 10, 11, 12, Table 10)

Etymology. From the type locality (Tahuata) Type locality. Matatehoke, Tahuata, Marquesas Island, French Polynesia. Material examined. Holotype: UFRJPOR 6454 = MNHN-IP-2018-25—Matatehoke, Tahuata, Marquesas Island, Station MT01 (9° 53.589’ S–149° 33.220’ W), depth: 33 m, coll. C. Debitus, 10/IX/2009, P177. Diagnosis. Sponge white, tubular, with cortical triactines, large and small choanosomal triactines, and choanosomal and atrial tetractines. Colour. White alive and beige in ethanol (Fig 10A). Morphology and anatomy. Sponge tubular with apical osculum (Figs 10A, B) surrounded by membrane (Fig 10C). Surface smooth. Large central atrium. Aquiferous system leuconoid. The cortical skeleton is composed of tangential triactines (Figs 11A, B). In the choanosome there are large triactines (larger than those of the cortex) and small choanosomal triactines and tetractines. Some of these small triactines and the small tetractines surround the choanosomal canals (Fig 10E). The subatrial skeleton is composed of triactines that point their unpaired actine to the cortex (Fig 10F). The atrial skeleton has tangential tetractines that point their apical actine into the atrium (Figs 10F, 11C, D). Spicules (Table 10) Cortical triactines. Subregular. Actines are slightly conical with sharp tips. Sometimes they are undulated. The unpaired actine is shorter than the paired ones (Fig 12A). Size: 449.0/ 32.6 µm (paired), 369.0/ 28.3 µm (unpaired). Choanosomal large triactines. Subregular. Actines are slightly conical with sharp tips. Sometimes they are undulated. The unpaired actine is shorter than the paired ones (Fig 12B). Size: 796.7/ 53.0 µm (paired), 537.8/ 50.3 µm (unpaired).

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 283 Choanosomal small triactines. Strongly sagittal, with curved paired actines when they are surrounding the canals. Actines are slightly conical with sharp tips. One of the paired actines is frequently shorter than the other actines (Fig 12C). Size: 151.4/ 11.0 µm (paired), 172.1/ 12.6 µm (unpaired).

FIGURE 10. Leucandra tahuatae sp. nov. (UFRJPOR 6454). A—Specimen in situ. B—Fixed specimen. C—Oscular membrane. D—Cross-section of the choanosome. E—Chonanosome showing a canal (black arrow: tetractine; white arrow: triactine). The asterisk indicates larvae. F—Subatrial (arrows) and atrial skeletons. Abbreviation: at = atrium; cx = cortex.

Choanosomal small tetractines. Strongly sagittal with curved paired actines, as they are find surrounding the canals. Actines are slightly conical with sharp tips. The unpaired actine can present the same length of the paired ones or be shorter. The paired actines are undulated. The apical actine is much shorter and thinner. It is conical, sharp an smooth (Fig 12D). Size: 158.1/ 11.7 µm (paired), 184.1/ 13.0 µm (unpaired), 50.9/ 8.9 µm (apical). Subatrial triactines. Strongly sagittal (T-shaped). Actines are slightly conical with sharp tips. The unpaired actine is longer than the paired ones (Fig 12E). Size: 163.6/ 13.2 µm (paired), 261.3/ 16.3 µm (unpaired). Atrial tetractines. Sagittal. Actines are slightly conical with sharp tips. The basal actines show the same length and they are straight. The apical actine is shorter than the basal ones, conical, sharp and smooth (Fig 12F). Size: 214.8/ 14.9 µm (paired), 215.5/ 14.8 µm (unpaired), 50.1/ 8.1 µm (apical). Ecology. There was algae inside the atrium of this sponge. Geographical distribution. Marquesas Island, French Polynesia (present work). Remarks. Most leucandras present diactines in their skeleton but L. tahuatae sp. nov. is part of a small group of this genus without diactines. Another characteristic that differentiates the new species from most leucandras is the presence of a subatrial skeleton. However, sometimes it is very difficult to be sure if a species does not have subatrial skeleton or if its author just did not mention it. Hence, to compare our new species other species of Leucandra, we considered only those with skeleton composed of cortical triactines, choanosomal large and small triactines, choanosomal tetractines, and atrial tetractines. We found only three most similar species to L. tahuatae sp. nov.: L. ramosa (Burton, 1934), L. mozambiquensis Van Soest & De Voogd, 2018, and L. pilula Van Soest & De Voogd, 2018.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 285 FIGURE 12. Leucandra tahuatae sp. nov. (UFRJPOR 6454). A—Cortical triactines. B—Choanosomal large triactines. C— Choanosomal small triactines. D—Choanosomal small tetractines. E—Subatrial triactines. F—Atrial tetractine.

Leucandra ramosa was originally described from Australia. It can be differentiated from L. tahuatae sp. nov. by its greyish-brown colour in ethanol (the new species is white) and by the size of some spicule categories. Corti- cal triactines: up to 210.0/ 11.0 (L. ramosa); 260.0˗449.0˗700.0/ 25.0˗32.6˗40.0—paired actine, 180.0˗369.0˗550.0/ 10.0˗28.3˗35.0—unpaired actine (L. tahuatae sp. nov.). Choanosomal large triactines: up to 960.0/ 64.0 (L. ramosa); 508.1˗796.7˗1016.1/ 37.8˗53.0˗64.9—paired actine, 378.4˗537.8˗756.7/ 37.8˗50.3˗54.1—unpaired actine (L. tahuatae sp. nov.). Choanosomal small triactines: up to 240.0/ 12.0 (L. ramosa); 108.0˗151.4˗202.5/ 9.5˗11.0˗13.5— paired actine, 129.6˗172.1˗221.4/ 10.8˗12.6˗13.5—unpaired actine (L. tahuatae sp. nov.). Leucandra mozambiquensis, recently described from Mozambique Channel, can be differentiated from the new species by its external morphology, as it is an “irregular cup-shaped hollow mass” (Van Soest & De Voogd 2018) while the new species is tubular, and mainly by the presence of tetractines instead of triactines in the subatrial skeleton. Leucandra pilula, described from Seychelles, is globose, while L. tahuatae sp. nov. is tubular. The for- mer has oxhorn-shaped cortical triactines, while ours has equiangular cortical triactines. Besides, some of their spicule categories have sizes. Cortical triactines: 216.0˗281.0˗372.0/ 16.0˗21.6˗28.0—paired actine, 178.0˗245.0˗326.0/ 15.0˗22.7˗31.0—unpaired actine (L. pilula); 260.0˗449.0˗700.0/ 25.0˗32.6˗40.0—paired actine, 180.0˗369.0˗550.0/ 10.0˗28.3˗35.0—unpaired actine (L. tahuatae sp. nov.). Subatrial triactines: 100.0˗186.0˗303.0/ 11.0˗16.6˗29.0—paired actine, 94.0˗172.0˗254.0/ 9.0˗17.8˗26.0—unpaired actine (L. pilula); 116.1˗163.8˗243.0/ 10.2˗13.2˗16.2—paired actine, 164.7˗261.3˗335.1/ 10.8˗16.3˗21.6—unpaired actine (L. tahuatae sp. nov.). Atrial tetractines: 136.0˗239.0˗380.0/ 9.0˗17.1˗32.0—paired actine, 101.0˗186.0˗271.0/ 14.0˗19.4˗32.0—unpaired ac-

286 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. tine, 45.0˗84.0˗130.0/ 4.0˗8.4˗11.0—apical actine (L. pilula); 155.0˗214.8˗325.0/ 13.5˗14.9˗15.0—paired actine, 150.0˗215.5˗300.0/ 12.5˗14.8˗15.0—unpaired actine, 30.0˗50.1˗67.5/ 6.3˗8.1˗10.0—apical actine (L. tahuatae sp. nov.)

TABLE 10. Spicule measurements of the holotype of Leucandra tahuatae sp. nov. (UFRJPOR 6454). Spicule Actine Length (µm) Width (µm) N min mean sd max min mean sd max Cortical triactine Paired 260.0 449.0 132.4 700.0 25.0 32.6 4.7 40.0 20 Unpaired 180.0 369.0 122.8 550.0 10.0 28.3 6.3 35.0 20 Choanosomal large Paired 508.1 796.7 149.2 1016.1 37.8 53.0 6.7 64.9 20 triactine Unpaired 378.4 537.8 122.1 756.7 37.8 50.3 5.3 54.1 20 Choanosomal small Paired 108.0 151.4 27.0 202.5 9.5 11.0 1.0 13.5 11 triactine Unpaired 129.6 172.1 28.7 221.4 10.8 12.6 1.2 13.5 11 Choanosomal tetrac- Paired 108.0 158.1 24.2 210.6 8.1 11.7 1.7 13.5 20 tine Unpaired 126.9 184.1 29.9 234.9 9.5 13.0 1.5 16.2 20 Apical 27.0 50.9 10.5 72.9 6.8 8.9 1.3 10.8 20 Subatrial triactine Paired 116.1 163.6 43.2 243.0 10.2 13.2 2.1 16.2 15 Unpaired 164.7 261.3 50.8 335.1 10.8 16.3 3.4 21.6 15 Atrial tetractine Paired 155.0 214.8 41.7 325.0 13.5 14.9 0.3 15.0 20 Unpaired 150.0 215.5 37.3 300.0 12.5 14.8 0.6 15.0 20 Apical 30.0 50.1 8.4 67.5 6.3 8.1 1.1 10.0 20

Molecular taxonomy

Calcinea ITS Both phylogenetic methods used, Bayesian Inference (BI) and Maximum Likehood (ML), recovered trees with similar topologies, as shown in Figure 13. In this tree we included Clathrina huahineae sp. nov., Ernstia variabilis sp. nov., and L. microraphis from French Polynesia. A sequence of L. chagosensis from French Polynesia was also included (BMOO16210), however, that sequence is not from one of our specimens. Clathrina was recovered as a monophyletic genus with high bootstrap (99%) and posterior probability (1.0). Clathrina huahineae sp. nov. was a sister species of C. mutabilis, clustering with a bootstrap of 100% and posterior probability of 1.0. The p-distance between them was of 3.6%. Ernstia was also recovered as monophyletic with high bootstrap (99%) and posterior probability (1.0) values. Se- quences belonging to Ernstia variabilis sp. nov. clustered into a highly supported clade (bootstrap: 100%; posterior probability: 1.0), with low p-distance (0–0.3%), which confirms morphological analysis of the specimens belonging to the same species. This clade was closely related to Ernstia citrea (bootstrap: 48%; posterior probability: 1.0), with a genetic divergence of 1.9% (p-distance), evidencing that these are two diferent species. Both E. variabilis sp. nov. and E. citrea formed a well supported clade with the E. pyrum (bootstrap: 100%; posterior probability: 1.0). As in previous studies the genus Leucetta was not monophyletic (e.g. Klautau et al. 2013). The specimen of Leucetta microraphis from French Polynesia grouped with other specimens of that species, with a bootstrap of 87% and posterior probability of 1.0. The intraspecific p-distance varied from 0 to 3.4%. Two subclades were formed, one including specimens from the Great Barrier Reef (GBR, Australia; bootstrap of 100%, posterior probability of 1.0) and another reuniting specimens from the Red Sea and our specimen from the French Polynesia (bootstrap of 91%, posterior probability of 1.0).

Calcinea C-LSU As for ITS, BI and ML analyses recovered trees with similar topologies as shown in Figure 14. In that tree we included four species from French Polynesia: Ernstia variabilis sp. nov., Leucascus simplex, Leucetta chagosensis, and L. microraphis.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 287 The genus Ernstia was also recovered as monophyletic with C-LSU, with high bootstrap (100%) and posterior probability (1.0) values. Ernstia variabilis sp. nov. presented an intraspecific p-distance of 0.2–0.7% and clustered into a big clade along with Ernstia adunca Fontana et al., 2018, Ernstia arabica Voigt et al., 2017, Ernstia klautauae Van Soest & De Voogd, 2015, Ernstia naturalis Van Soest & De Voogd, 2015 and Ernstia aff. naturalis (bootstrap: 88%; posterior probability: 1.0). However, relationship among these species was not highly supported and topology differences were obtained in ML and Bayesian methods. This is the first time that a C-LSU sequence of the type species of Leucascus, L. simplex Dendy, 1892, was generated. Despite this, the three species of Leucascus included in our tree grouped with 98% bootstrap and 1.0 of posterior probability. Leucascus simplex from French Polynesia presented an intraspecific p-distance of 0.5% and was sister species of L. flavus, with a p-distance of 2.0%. Like in the ITS phylogeny, the clade of Leucetta is polyphyletic and falls in a clade with Ascoleucetta, Leucet- tusa and Pericharax. Our specimens of L. chagosensis and L. microraphis grouped inside this major Leucetta clade. Leucetta chagosensis formed a clade with specimens from French Polynesia, Coral Sea, and Indonesia (66% bootstrap, 1.0 of posterior probability). The intraspecific p-distance in this clade varied from 0.0 to 2.2%. Leucetta microraphis formed a more intricate clade, as it included L. sulcata (72% bootstrap, 1.0 of posterior probability). We can recognise six subclades in the L. microraphis sensu lato clade: one reuniting Leucetta sulcata specimens (87% bootstrap, 1.0 of posterior probability), a second clade with specimens from French Polynesia (63% bootstrap, 1.0 of posterior probability), a third one reuniting specimens from Rodrigues (98% bootstrap, 1.0 of posterior probability). The two specimens from the Red Sea did not form a clade. In fact, one of these specimens grouped with the Indonesian specimen with low support (<50% bootstrap, 0.6 of posterior probability). Finally, the specimens from the Great Barrier Reef (Australia) grouped with high support (80% bootstrap, 1.0 of posterior probability). Leucetta sulcata was considered a species different from L. microraphis because of its white colour and “grooved and holed habitus” (Van Soest & De Voogd, 2018). In fact, analysing the external morphology of the specimens in this clade they present different habitus.

Calcaronea C-LSU Both BI and ML trees presented similar topologies. The ML tree is shown in Figure 15 and both bootstrap and posterior probability supports are presented there. In this tree, the only species from the French Polynesia is Leucandra tahuatae sp. nov. Leucandra is a genus al- ready known as being paraphyletic (e.g. Voigt et al. 2012a; Klautau et al. 2016; Alvizu et al. 2018; Cóndor-Luján et al. 2018). In our tree, Leucandra showed again to be paraphyletic. Our new species, Leucandra tahuatae, was sister species of a large clade composed of Leucandra and Paraleucilla Dendy, 1892 species and of Breitfussia schulzei (Breitfuss, 1896). Two species morphologically similar to L. tahuatae sp. nov. were included in the tree: L. mozambiquensis and L. pilula. The new species presented a p-distance of 4.4% with L. mozambiquensis and of 4.3% with L. pilula.

Discussion and conclusion

With the present work, the number of calcareous sponges from French Polynesia increased from 6 to 15. Consider- ing just calcareous sponges, we re-collected only one (Leucetta chagosensis) of the five previously known species. The other seven species are new occurrences in French Polynesia and six of them are new to science. It is also the first time that the genera Ascandra, Clathrina, Ernstia, Leucascus, and Leucandra were found in French Polynesia. According to our results, the Eastern Indo-Pacific Realm shows more affinity with the Central and the Western Indo-Pacific Realms. The three species that support these affinities are Ascandra crewsi, previously known only from Papua New Guinea (Van Soest & De Voogd 2015), and Leucetta chagosensis and L. microraphis, both widespread species in the Indo-Pacific. The high intraspecific p-distance of L. microraphis and of L. chagosensis suggest that these species can represent species complexes or that they have a complex evolutionary history of founder and vicariant events that resulted in regional isolation and incipient speciation processes (Wörheide et al. 2008; Voigt et al. 2012, Van Soest & De Voogd 2018).

288 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. FIGURE 13. ML phylogenetic tree of the ITS from Calcinea. Species sequences found in the present work are highlighted. Support values are shown at the nodes (Posterior Probability and Bootstrap values; < means value below 50%,—means node not present in the Bayesian phylogeny). Midpoint rooted tree.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 289 FIGURE 14. ML phylogenetic tree of the LSU from Calcinea. Species sequences found in the present work are highlighted. Support values are shown at the nodes (Posterior Probability and Bootstrap values; < means value below 50%,—means node not present in the Bayesian phylogeny). Midpointed root tree.

290 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. FIGURE 15. ML phylogenetic tree of the LSU from Calcaronea. Species sequences found in the present work are highlighted. Support values are shown at the nodes (Posterior Probability and Bootstrap values; < means value below 50%,—means node not present in the Bayesian phylogeny). Tree rooted with Leucosolenia spp.

For L. microraphis, however, at the moment is very difficult to recognise which clade could contain the true L. microraphis. This species was a variety of L. primigenia Haeckel, 1872 and, to our knowledge, no type nor type locality was ellected by its author. Therefore, until a revision of the genus with perhaps the proposal of a neotype is done, it will be very difficult to unravel this species complex. The molecular markers ITS and C-LSU once again helped the identification of calcareous sponges, showing how important is an integrative taxonomy. The increase of 250% (6 spp to 15 spp) shows how poorly known is the diversity of Calcarea in the French Polynesia. More studies on this group are desirable not only to know the true biodiversity of Calcarea in French Polynesia, but to help understanding the evolutionary history of Calcarea in the world.

We thank French Polynesian authorities for permitting and kindly support our sponge surveys in French Polynesia, IRD for funding of R/V Alis field trips, R/V Alis crew and the IRD-Noumea diving team for their help. This work was funded by the Brazilian National Research Council (CNPq) and by the Coordination for the Improvement of Higher Education Personnel (CAPES). M.K. received a fellowship from CNPq. M.V.L. and B.G. received scholar- ships from CAPES (PROTAX) and PIBIC/UFRJ, respectively.

References

Azevedo, F., Cóndor-Luján, B., Willenz, P., Hajdu, E., Hooker, Y. & Klautau, M. (2015) Integrative taxonomy of calcareous sponges (subclass Calcinea) from the Peruvian coast: morphology, molecules, and biogeography. Zoological Journal of the Linnean Society, 173, 787–817. https://doi.org/10.1111/zoj.12213 Azevedo, F., Galinou-Mitsoudi, S. & Gerovasileiou, V. (2017a) First record of the invasive sponge Paraleucilla magna (Po- rifera, Calcarea) in Greek waters. In: Gerovasileiou, V., Akel, E.H.Kh., Akyol, O. & Alongi, G. (Eds.), New Mediterranean Biodiversity Records. Mediterranean Marine Science, 18 (1), pp. 368–369. https://doi.org/10.12681/mms.2068 Azevedo, F., Padua, A., Moraes, F., Rossi, A., Muricy, G. & Klautau, M. (2017b) Taxonomy and phylogeny of calcareous sponges (Porifera: Calcarea: Calcinea) from Brazilian mid-shelf and oceanic islands. Zootaxa, 4311 (3), 301–344. https://doi.org/10.11646/zootaxa.4311.3.1 Baine, M. & Harasti, D. (2007) The marine life of Bootless Bay, Papua New Guinea. Motupore Island Research Centre, Papua New Guinea, pp. 1–144. Bidder, G.P. (1898) The skeleton and the classification of calcareous sponges. Proceedings of the Royal Society of London, 64, 61–76. https://doi.org/10.1098/rspl.1898.0070 Borojević, R. (1966). Éponges calcaires des côtes de France. II. Le genre Ascandra Haeckel emend. Archives de Zoologie Ex- périmentale et Générale, 107 (2), 357–367. Borojević, R. (1967) Spongiaires d’Afrique du Sud. (2) Calcarea. Transactions of the Royal Society of South Africa, 37 (3), 183–226. https://doi.org/10.1080/00359196709519066 Borojević, R. (1971) Eponges calcaires des côtes du Sud-Est du Brésil, épibiontes sur Laminaria brasiliensis et Sargassum cymosum. Revista Brasileira de Biologia, 31, 525–530. Borojević, R. & Boury-Esnault, N. (1987) Revision of the genus Leucilla Haeckel, 1872, with a re-description of the type species—Leucilla amphora Haeckel, 1872. In: Jones, W.C. (Ed.), European contributions to the taxonomy of sponges. Sherkin Island Marine Station, Sherkin Island, County Cork, pp. 29–40. Borojević, R. & Klautau, M. (2000) Calcareous sponges from New Caledonia. Zoosystema, 22 (2), 187–201. Borojević, R. & Peixinho, S. (1976) Éponges calcaires des côtes nord et nord-est du Brésil. Bulletin du Muséum d’histoire Na- turelle de Paris, 402, 987–1036. Bowerbank, J.S. (1862) On the Anatomy and Physiology of the Spongiadae. Part II. Philosophical Transactions of the Royal Society, 152, 747–829. https://doi.org/10.1098/rstl.1862.0035 Bowerbank, J.S. (1866) A Monograph of the British Spongiadae. Vol. 2. Ray Society, London, xx + 388 pp. Available from: https://www.biodiversitylibrary.org/item/18176#page/7/mode/1up (accessed 12 January 2020) Breitfuss, L.L. (1896) Kalkschwämme von Ternate Molukken), nach den Sammlungen Prf. W. Kükenthal’s (Vorläufige Mit- theilung). Zoologischer Anzeiger, 19, 433–435. Breitfuss, L.L. (1898) Kalkschwämme von Ternate. Abhandlungen der Senckenbergischen Naturforschender Gesellschaft, 24, 169–178. Burton, M. (1934) Sponges. Great Barrier reef Expedition, 1928–29, 4 (14), 513–621. Burton, M. (1963) Revision of the classification of the calcareous sponges. British Museum (Natural History), London, 693 pp Carter, H.J. (1886) Descriptions of the sponges from the neighbourhood of Port Philip Heads, South Australia. Annals and Magazine of Natural History, 15–18, 431–441. https://doi.org/10.1080/00222938609460169 Cavalcanti, F.F.; Rapp, H.T & Klautau, M. (2013) Taxonomic revision of Leucascus Dendy, 1892 (Porifera: Calcarea) with re- validation of Ascoleucetta Dendy & Frederick, 1924 and description of three new species. Zootaxa, 3619 (3), 275–314. https://doi.org/10.11646/zootaxa.3619.3.3 Cavalcanti, F.F., Skinner, L.F. & Klautau, M. (2013) Population dynamics of cryptogenic calcarean sponges (Porifera, Calcarea) in Southeastern Brazil. Marine Ecology, 34, 280–288. https://doi.org/10.1111/maec.12013

292 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. Chombard, C., Boury-Esnault, N. & Tillier, S. (1998) Reassessment of homology of morphological characters in tetractinellid sponges based on molecular data. Systematic Biology, 47, 351–366. https://doi.org/10.1080/106351598260761 Colin, P.L. & Arneson, C. (1995) Tropical Pacific Invertebrates. A field guide to the Marine Invertebrates occurring on Tropical Pacific Coral Reefs, Seagrass Beds and Mangroves. Coral Reef Press, Irvine, pp. 1–296. Cóndor-Luján, B., Louzada, T.S., Hajdu, E. & Klautau, M. (2018) Morphological and molecular taxonomy of calcareous sponges (Porifera: Calcarea) from Curaçao, Caribbean Sea. Zoological Journal of the Linnean Society, 182 (1), 1–67. https://doi.org/10.1093/zoolinnean/zlx082 Debitus, C. (2009) BSMPF-1 cruise. Alis R/V. https://doi.org/10.17600/9100030 Debitus, C. (2011) TUAM’2011 cruise. Alis R/V. https://doi.org/10.17600/11100010 Debitus, C. (2013a) Tuhaa PAE 2013 cruise. Alis R/V. https://doi.org/10.17600/13100030 Debitus, C. (2013b) Tahiti ITI cruise. Alis R/V. https://doi.org/10.17600/13100040 De Laubenfels, M.W. (1936) A Discussion of the Sponge Fauna of the Dry Tortugas in Particular and the West Indies in General, with Material for a Revision of the Families and Orders of the Porifera. Carnegie Institute of Washington Publication, 467, 1–225. [Tortugas Laboratory Paper 30] Dendy, A. (1892) Synopsis of the Australian Calcarea Heterocoela; with a proposed classification of the group and descriptions of some new genera and species. Proceedings of the Royal Society of Victoria, 5, 69–116. Dendy, A. (1913) The Percy Sladen trust expedition to the Indian Ocean in1905 (V). I. Report on the Calcareous Sponges collected by HMS ‘Sealark’ in the Indian Ocean. Transactions of the Linnean Society of London, Series 2, 16 (1), 1–29. https://doi.org/10.1111/j.1096-3642.1914.tb00121.x Dendy, A. & Frederick, L.M. (1924) On a collection of sponges from the Abrolhos Islands, Western Australia. Journal of the Linnean Society of London, Zoology, 16, 1–29. https://doi.org/10.1111/j.1096-3642.1924.tb00052.x Dendy, A. & Row, R.W.H. (1913) The classification and phylogeny of the Calcareous Sponges, with a reference list of all the described species, systematically arranged. Proceedings of the Zoological Society of London, 1913 (3), 704–813. https://doi.org/10.1111/j.1469-7998.1913.tb06152.x Erhardt, H. & Baensch, H.A. (1998) Meerwasser Atlas 4. Wirbellose. Mergus Verlag, Melle, 1214 pp. Fontana, T., Cóndor-Luján, B., Azevedo, F., Pérez, T. & Klautau, M. (2018) Diversity and distribution of Calcareous sponges (subclass Calcinea) from Martinique. Zootaxa, 4410 (2), 331–369. https://doi.org/10.11646/zootaxa.4410.2.5 Gosliner, T.M., Behrens, D.W & Williams, G.C. (1996) Coral reef animals of the Indo-Pacific: animal life from Africa to Hawaii exclusive of the vertebrates. Sea Challengers, Monterey, 314 pp. Gray, J.E. (1867) Notes on the arrangement of sponges, with the descriptions of some new genera. Proceedings of the Zoological Society of London, 1867 (2), 492–558. [https://www.biodiversitylibrary.org/item/93424#page/ 514/mode/1up] Haeckel, E. (1872) Die Kalkschwämme, eine Monographie. G. Reimer, Berlin, 512 + 440 + 260 pp. https://doi.org/10.5962/bhl.title.11323 Hall, K.A., Sutcliffe, P.R, Hooper, J.N.A., Alencar, A., Vacelet, J., Pisera, A., Petek, S., Folcher, E., Butscher, J., Orempuller, J., Maihota, N. & Debitus, C. (2013) Affinities of Sponges (Porifera) of the Marquesas and Society Islands, French Polynesia. Pacific Science, 67 (4), 493–511. https://doi.org/10.2984/67.4.1 Hartman, W. (1958) A re-examination of Bidder’s classification of the Calcarea. Systematic Zoology, 7, 97–110. https://doi.org/10.2307/2411971 Huelsenbeck, J.P. & Ronquist, F. (2001) MRBAYES: Bayesian inference of phylogeny. Bioinformatics, 17, 754–755 https://doi.org/10.1093/bioinformatics/17.8.754 Jenkin, C.F. (1908) The marine fauna of Zanzibar and British East Africa, from collections made by Cyril Crossland, M.A., in the years 1901 & 1902. The Calcareous Sponges. Proceedings of the Zoological Society of London, 1908, 434–456. [https:// www.biodiversitylibrary.org/item/99643#page/46/mode/1up] https://doi.org/10.1111/j.1469-7998.1908.tb07387.x Katoh, K. & Standley, D.M. (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution, 30, 772–780. https://doi.org/10.1093/molbev/mst010 Katoh, K. & Toh, H. (2008) Improved accuracy of multiple ncRNA alignment by incorporating structural information into a MAFFT-based framework. BMC Bioinformatics, 9, 212. https://doi.org/10.1186/1471-2105-9-212 Kelly-Borges, M. & Valentine, C. (1995) The Sponges of the Tropical Island Region of Oceania: A Taxonomic Status Review. In: Maragos, J.E., Peterson, M.N.A., Eldredge, L.G., Bardach, J.E. & Takeuchi, H.F. (Eds.), Marine and Coastal Biodiver- sity in the Tropical Island Pacific Region. Vol. 1. species systematics and information management priorities. East-West Center, University of Hawaii, Honolulu, pp. 83–120.

Calcarea from French Polynesia Zootaxa 4748 (2) © 2020 Magnolia Press · 293 Kirkpatrick, R. (1910) On a Remarkable Pharetronid Sponge from Christmas Island. Proceedings of the Royal Society (B), 83 (562), 124–133. https://doi.org/10.1098/rspb.1910.0070 Klautau, M., Azevedo, F., Cóndor-Luján, B., Rapp, H.T., Collins, A. & Russo, C.A.M. (2013) A molecular phylogeny for the Order Clathrinida rekindles and refines Haeckel’s taxonomic proposal for calcareous sponges. Integrative and Compara- tive Biology, 53, 447–461. https://doi.org/10.1093/icb/ict039 Klautau, M. & Borojević, R. (2001) Sponges of the genus Clathrina from Arraial do Cabo, Brazil. Zoosystema, 23, 395–410. Klautau, M., Imesek, M., Azevedo, F.C., Plese, B., Nikolić, V. & Cetković, H. (2016) Adriatic calcarean sponges (Porifera, Cal- carea) with description of six new species and richness analysis. European Journal of Taxonomy, 178, 1–52. https://doi.org/10.5852/ejt.2016.178 Klautau, M., Monteiro, L. & Borojević, R. (2004) First occurrence of the genus Paraleucilla (Calcarea, Porifera) in the Atlantic Ocean: P. magna sp. nov. Zootaxa, 710 (1), 1–8. https://doi.org/10.11646/zootaxa.710.1.1 Klautau, M. & Valentine, C. (2003) Revision of the genus Clathrina (Porifera, Calcarea). Zoological Journal of the Linnean Society, 139, 1–62. https://doi.org/10.1046/j.0024-4082.2003.00063.x Lanna, E. & Klautau, M. (2016) Life history and reproductive dynamics of the cryptogenic calcareous sponge Sycettusa has- tifera (Porifera, Calcarea) living in tropical rocky shores. Journal of the Marine Biological Association of the United Kingdom, 1, 1–10. https://doi.org/10.1017/S0025315416001466 Lanna, E., Rossi, A.L., Cavalcanti, F.F., Hajdu, E. & Klautau, M. (2007) Calcareous sponges from São Paulo state, Brazil (Po- rifera: Calcarea: Calcinea) with the description of two new species. Journal of the Marine Biological Association of the United Kingdom, 87, 1553–1561. https://doi.org/10.1017/S0025315407056871 Lévi, C., Laboute, P., Bargibant, G. & Menou, J.L. (Eds.) (1998) Sponges of the New Caledonian Lagoon. Éditions ORSTOM, Paris, 211 pp. Lôbo-Hajdu, G., Guimarães, A., Salgado, A., Lamarão, F., Vieiralves, T., Mansure, J. & Albano, R. (2004) Intragenomic, in- tra- and interspecific variation in the rDNA ITS of a Porifera revealed by PCR-single-strand conformation polymorphism (PCR-SSCP). Bollettino dei musei e degli istituti biologici dell’Universita di Genova, 68, 413–423. Longo, C., Mastrototaro, F. & Corriero, G. (2007) Occurrence of Paraleucilla magna (Porifera: Calcarea) in the Mediterranean Sea. Journal of the Marine Biological Association of the United Kingdom, 87, 1749–1755. https://doi.org/10.1017/S0025315407057748 Mačić, V. & Petović, S. (2017) New data on the distribution of the alien sponge Paraleucilla magna Klautau, Monteiro & Borojević, 2004 in the Adriatic Sea. Studia Marina, 29 (1), 63–68. Minchin, E.A. (1900) Chapter III. Sponges. In: Lankester, E.R. (Ed.), A Treatise on Zoology. Part II. The Porifera and Coelen- terata. 2. Adam & Charles Black, London, pp. 1–178. Moraes, F.C., Ventura, M., Klautau, M., Hajdu, E. & Muricy, G. (2006) Biodiversidade de esponjas das ilhas oceânicas brasileiras. In: Alves, R.V. & Castro, J.W. (Ed.), Ilhas oceânicas brasileiras—da pesquisa ao manejo. Ministério do Meio Ambi- ente, Brasília, pp. 147–177. Moraes, F.C., Vilanova, E.P. & Muricy, G. (2003) Distribuição das Esponjas (Porifera) na Reserva Biológica do Atol das Rocas, Nordeste do Brasil. Arquivos do Museu Nacional, 61, 13–22. Nei, M. & Kumar, S. (2000) Molecular Evolution and Phylogenetics. Oxford University Press, New York, 352 pp. Padua, A. & Klautau, M. (2016) Regeneration in calcareous sponges (Porifera). Journal of the Marine Biological Association of the United Kingdom, 96, 553–558. https://doi.org/10.1017/S0025315414002136 Petek, S. & Debitus, C. (2017) Sponges of Polynesia [on line]. IRD, Papeete (PYF), 827 pp. Available from: https://sponges- polynesia.ird.fr/ (accessed 12 January 2020) Poléjaeff, N. (1883) Report on the Calcarea dredged by H.M.S.‘Challenger’, during the years 1873-1876. Report on the Scien- tific Results of the Voyage of H.M.S. ‘Challenger’, 1873–1876, Zoology, 8 (2), 1–76. Pulitzer-Finali, G. (1982 [1980-1981]) Some new or little-known sponges from the Great Barrier Reef of Australia. Bollettino dei Musei e degli Istituti Biologici dell’Universitá di Genova, 48–49, 87–141. Rapp, H.T. (2004) The first record of the genus Leucascus Dendy, 1892 from the Atlantic Ocean, with description of Leucascus lobatus sp. nov. (Porifera, Calcarea, Calcinea) from Greenland. Steenstrupia, 28 (2), 1–9. Rapp, H.T. (2006). Calcareous sponges of the genera Clathrina and Guancha (Calcinea, Calcarea, Porifera) of Norway (north- east Atlantic) with the description of five new species. Zoological Journal of the Linnean Society, 147 (3), 331–365. https://doi.org/10.1111/j.1096-3642.2006.00221.x Rapp, H.T., Göcke, C., Tendal, O.S. & Janussen, D. (2013) Two new species of calcareous sponges (Porifera: Calcarea) from the deep Antarctic Eckström Shelf and a revised list of species found in Antarctic waters. Zootaxa, 3692 (1), 149–159. https://doi.org/10.11646/zootaxa.3692.1.9 Ridley, S.O. (1884) Spongiida. In: Report on the Zoological Collections made in the Indo-Pacific Ocean during the Voyage of H.M.S. ‘Alert’, 1881–2. British Museum (Natural History), London), pp. 366–482 + 582–630.

294 · Zootaxa 4748 (2) © 2020 Magnolia Press Klautau et al. Ronquist, F. & Huelsenback, J.P. (2003) MRBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19, 1572–1574. https://doi.org/10.1093/bioinformatics/btg180 Row, R.W.H. (1909) Reports on the marine biology of the Sudanese Red Sea. XIII. Report on the Sponges, collected by Mr. Cyril Crossland in 1904–5. Part I. Calcarea. Journal of the Linnean Society. Zoology, 31 (206), 182–214. [https://academic. oup.com/zoolinnean/article/31/206/182/2682838] https://doi.org/10.1111/j.1096-3642.1909.tb00983.x Row, R.W.H. & Hôzawa, S. (1931) Report on the Calcarea obtained by the Hamburg South-West Australian Expedition of 1905. Science Reports of the Tôhoku University, Series 4, 6 (1), 727–809. Salvat, B., Aubanel, A., Adjeroud, M., Bouisset, P., Calmet, D., Chancerelle, Y., Cochennec, N., Davies, N., Fougerouse, A., Galzin, R., Lagouy, E., Lo, C., Monier, C., Ponsonnet, C., Remoissenet, G., Schneider, D., Stein, A., Tatarata, M. & Villiers, L. (2008) Le suivi de l’état de santé des récifs coralliens de Polynésie française et leur récente évolution. Revue d’écologie, Terre et Vie, 62, 145–177. Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning. A laboratory Manual. Harbor Laboratory Press, Cold Spring, New York, 1626 pp. Solé-Cava, A.M., Klautau, M., Boury-Esnault, N., Borojević, R. & Thorpe, J.P. (1991) Genetic evidence for cryptic speciation in allopatric populations of two cosmopolitan species of the calcareous sponge genus Clathrina. Marine Biology, 111, 381–386. https://doi.org/10.1007/BF01319410 Tanita, S. (1942) Report on the Calcareous sponges obtained by the Zoological Institute and Museum of Hamburg. Part II. Sci- ence Reports of the Tôhoku University, Series 4, 17 (2), 105–135. Thacker, A.G. (1908) On collections of the Cape Verde Islands fauna made by Cyril Crossland, M.A. The Calcareous sponges. Proceedings of the Zoological Society of London, 49, 757–782. Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013) MEGA6: Molecular Evolutionary Genetics Analysis Ver- sion 6.0. Molecular Biology and Evolution, 30, 2725–2729. https://doi.org/10.1093/molbev/mst197 Topaloðlu, B., Evcen, A., Cinar, M.E., 2016. Sponge fauna in the Sea of Marmara. Turkish Journal of Fisheries and Aquatic Sciences, 16 (1), 51–59. https://doi.org/10.4194/1303-2712-v16_1_06 Vacelet, J. (1967) Description d’éponges Pharétronides actuelles des tunnels obscurs sous-récifaux de Tuléar (Madagascar). Re- cueil des Travaux de la Station marine d’Endoume, Fascicule Hors Série, 6, 37–62. Vacelet, J. (1977) Éponges pharétronides actuelles et sclérosponges de Polynésie française, de Madagascar et de la Réunion. Bulletin du Muséum National d’histoire Naturelle de Paris, 444, 345–368. Van Soest, R.W.N. & De Voogd, N.J. (2015) Calcareous sponges of Indonesia. Zootaxa, 3951 (1), 1–105. https://doi.org/10.11646/zootaxa.3951.1.1 Van Soest, R.W.N. & De Voogd, N.J. (2018) Calcareous sponges of the Western Indian Ocean and Red Sea. Zootaxa, 4426 (1), 1–160. https://doi.org/10.11646/zootaxa.4426.1.1 Voigt, O., Eichmann, V. & Wörheide, G. (2012b) First evaluation of mitochondrial DNA as a marker for phylogeographic studies of Calcarea: a case study from Leucetta chagosensis. Hydrobiologia, 687, 101–106. https://doi.org/10.1007/s10750-011-0800-7 Voigt, O. & Wörheide, G. (2016) A short LSU rRNA fragment as a standard marker for integrative taxonomy in calcareous sponges (Porifera: Calcarea). Organisms Diversity & Evolution, 16, 53–64. https://doi.org/10.1007/s13127-015-0247-1 Voigt, O., Wülfingl, E. & Wörheide, G. (2012a) Molecular Phylogenetic Evaluation of Classification and Scenarios of Character Evolution in Calcareous Sponges (Porifera, Class Calcarea). PLoS ONE, 7, e33417. https://doi.org/10.1371/journal.pone.0033417 Von Lendenfeld, R. (1885) A Monograph of the Australian Sponges (Continued). Part III. Preliminary description and classification of the Australian Calcispongiae. Proceedings of the Linnean Society of New South Wales, 9, 1083–1150. Wörheide, G., Epp, L.S. & Macis, L. (2008) Deep genetic divergences among Indo-Pacific populations of the coral reef sponge Leucetta chagosensis (Leucettidae): Founder effects, vicariance, or both? BMC Evolutionary Biology, 8, 24. https://doi.org/10.1186/1471-2148-8-24 Wörheide, G. & Hooper, J.N.A. (1999) Calcarea from the Great Barrier Reef. 1: Cryptic Calcinea from Heron Island and Wistari Reef (Capricorn-Bunker Group). Memoirs of the Queensland Museum, 43, 859–891. Wörheide, G., Hooper, J.N.A. & Degnan, B.M. (2002) Phylogeography of Western Pacific Leucetta ‘chagosensis’ (Porifera: Calcarea) from ribosomal DNA sequences: implications for population history and conservation of the Great Barrier Reef World Heritage Area (Australia). Molecular Ecology, 11, 1753–1768. https://doi.org/10.1046/j.1365-294X.2002.01570.x Wörheide, G., Solé-Cava, A.M. & Hooper, J.N.A. (2005) Biodiversity, molecular ecology and phylogeography of marine sponges: patterns, implications and outlooks. Integrative and Comparative Biology, 45, 377–385. https://doi.org/10.1093/icb/45.2.377