New records of Neopetrosia carbonaria (Lamarck, 1814) from the Brazilian coast reveal new morphological features and spicule types Author Sandes, Joana 0000-0003-0311-8250 Museu Nacional, Departamento de Invertebrados, Universidade Federal do Rio de Janeiro, Quinta da Boa Vista, s / n, CEP 20940 - 040, Rio de Janeiro, RJ, Brazil. & https: // orcid. org / 0000 - 0003 - 0311 - 8250 Author Rocha, Lívia 0000-0002-5284-1309 Museu Nacional, Departamento de Invertebrados, Universidade Federal do Rio de Janeiro, Quinta da Boa Vista, s / n, CEP 20940 - 040, Rio de Janeiro, RJ, Brazil. & https: // orcid. org / 0000 - 0002 - 5284 - 1309 Author Pinheiro, Ulisses 0000-0003-3658-1372 Universidade Federal de Pernambuco, Centro de Ciências Biológicas, Departamento de Zoologia, Av. Nelson Chaves, s / n, Cidade Universitária CEP 50373 - 970, Recife, PE, Brazil. https: // orcid. org / 0000 - 0003 - 3658 - 1372 Author Santos, George Garcia 0000-0002-0785-9913 Universidade Federal do Cariri, Instituto de Formação de Educadores, Rua Olegário Emídio de Araújo, s / n, CEP 63260 - 000, Brejo Santo, CE, Brazil. https: // orcid. org / 0000 - 0002 - 0785 - 9913 text Zootaxa 2023 2023-04-14 5264 3 429 439 http://dx.doi.org/10.11646/zootaxa.5264.3.10 journal article 249447 10.11646/zootaxa.5264.3.10 f6a37c5d-38a0-4e05-8438-eab5f04b550d 1175-5326 7836620 12D73CA9-7609-4DDE-8C19-3C69E26E45B7 Neopetrosia carbonaria (Lamarck, 1814) ( Figures 2–4 , Table 1 ) Spongia carbonaria Lamarck, 1814: 375 . Thalysias carbonaria Duchassaing & Michelotti, 1864: 83–84 . Pachychalina carbonaria Arndt, 1927: 152–153 . Phloeodictyon carbonaria Topsent, 1930: 26–27 . Adocia carbonaria Hechtel, 1965: 26 ; Hechtel, 1969: 9 ; Hechtel, 1976: 252 (non Adocia carbonaria sensu Rocha, 1995 [= Haliclona ( Soestella ) melana ]). Pellina carbonaria De Laubenfels, 1936: 68–69 ; Van Soest, 1980: 83 ; Pulitzer-Finali, 1986: 159 ; Zea, 1987: 125 . Xestospogia carbonaria Lehnert & Van Soest, 1998: 93 ; Ŗtzler et al. , 2000: 238; Díaz, 2005: 470 . Neopetrosia carbonaria Ŗtzler et al. , 2009: 304; Zea et al. , 2014 ; Pérez et al. , 2017: 10 . Neopetrosia aff. carbonaria Rocha et al. , 2021: 262 . Collected material ( six specimens ). Northeastern Brazil , Paraíba State : UFPEPOR 1637, Carapibus beach ( 7°17′57.66″ S , 34°47′52.93″ W ), Paraíba State , Brazil, 1 m depth , coll. G. G. Santos , 31 January 2014 ; UFPEPOR 1853, 1854, 1855, Carapibus beach ( 7°17′57.66″ S , 34°47′52.93″ W ), Paraíba State , Brazil , 1 m depth , coll. G. G. Santos , 21 February 2015 . Northeastern Brazil , Pernambuco State : UFPEPOR 1458, 1472, Enseada dos Corais beach ( 8°19′23″ S , 34°56′57″ W ), Cabo de Santo Agostinho Municipality , Pernambuco State , Brazil, 1 m depth , colls. U. Pinheiro and G. G. Santos , 26 November 2012 . Additional examined material ( 15 specimens ) . Northeastern Brazil , Ceará State : MNRJ 17400 , Icaraizinho de Amontada , intertidal, coll. Rômulo Farias Kyria Santiago , November 2013 ; MNRJ 17866 , Mundaú beach ( 3.17418º S , 39.35723º W ), Trairí city, 1 m depth, coll. M. Carvalho, 29 March 2014 ; MNRJ 17741 , MNRJ 17766 , Mundaú beach ( 3.17418º S , 39.35723º W ), Trairí city, 1 m depth , coll. E. Hajdu , 29 and 31 March 2014 ; MNRJ 17777 , Mundaú beach ( 3.17418º S , 39.35723º W ), Trairí city, 1 m depth , coll. A. Bispo , 29 March 2014 ; MNRJ 17832 , Mundaú beach ( 3.17418º S , 39.35723º W ), Trairí city, 1 m depth , coll. S. Salani , 29 March 2014 ; MNRJ 17782 , 17783 , Flecheiras beach ( 3.21858º S , 39.26344º W ), 1 m depth , coll. A. Bispo , 30 March 2014 ; MNRJ 17840 , Flecheiras beach ( 3.21858º S , 39.26344º W ), 1 m depth , coll. S. Salani , 30 March 2014 ; MNRJ 17887 , Flecheiras beach ( 3.21858º S , 39.26344º W ), 1 m depth , coll. M. Carvalho , 30 March 2014 . Northeastern Brazil , Rio Grande do Norte State : MNRJ 18033 , Parrachos de Maracajaú , Maxaranguape City ( 5.40114º S , 35.29677º W ), depth not recorded, coll. S. Salani , 6 April 2014 . Northeastern Brazil , Bahia State : MNRJ 18690 , Abrolhos Bank , Pedra de Leste , 3 m depth , coll. F. Moraes , 30 June 2014 . Northeastern Brazil , Alagoas State : MNRJ 4713 , Ponta Verde beach ( 9.65000º S , 35.68333º W ), Maceió city, 0.5–1 m depth , coll. E. Hajdu , 3 September 2001 ; MNRJ 17035 , Ponta do Meirim beach ( 9.70000º S , 35.76667º W ), Maceió city, 0.5–1 m depth , coll. A. Bispo , 10 January 2012 . Comparative material. Spongia carbonaria Lamarck, 1814 , Holotype MNHN-LBIM-DT 545, Schyzotype MNRJ 21834. Adocia carbonaria Hetchel, 1976 , YPM IZ 8960, Piedade reef, near Recife city, Pernambuco State , Brazil , coll. Jacques Laborel, 21 November 1961 . Neopetrosia carbonaria , MNRJ 9405, Smithsonian Tropical Research Institute, Bocas del Toro, 1 m depth, coll. G. Muricy, 15 August 2005 . New diagnosis. Neopetrosia carbonaria is a thick encrusting to repent or ramose sponge, with reddish brown to brown color or dark green to black color in vivo , two categories of oxeas as megascleres and rare raphidiform toxas as microscleres. External morphology ( Fig. 2 ). Thickly encrusting ( Fig. 2A–D ) to repent ( Fig. 2E–F ) or ramose growth form ( Fig. 2G–H ), up to 7.5 x 3.5 x 1.5 cm (length x width x thickness). The surface is smooth to rough, uneven, easily detachable. The subdermal spaces are visible. The oscules have a lighter-colored rim and may be flush with the surface or raised on volcano-like to tubular projections up to 1 cm high, 0.5–4.0 mm in diameter. The consistency is hard and crumbly. The color in vivo varies from reddish brown ( Figs. 2A–B and 2E–F ) to brown ( Fig. 2G–H ) or dark green ( Fig. 2C ) to almost black ( Fig. 2D ) and becomes dark brown or black, when preserved. The preservation alcohol turns bluish or greenish black when the sponge is placed in it, becoming black after a while. Skeleton ( Fig. 3 ). The ectosomal skeleton is a tangential isodictyal reticulation of single oxeas I, overlying a subsuperficial reticulation. The dermal membrane is pierced by pores with 17–38 μm in diameter. Spongin is abundant inside the ectosomal meshes and at the nodes of spicules in the isodictyal ectosome ( Fig. 3A–B ). Short pauci- to multispicular perpendicular tracts support the surface and form the regularly distributed subdermal spaces, with 200–1.000 μm in diameter ( Fig. 3C ). The choanosome is a dense mass of oxeas I forming vague, rounded meshes (100–800 μm in diameter), which are largely obscured by longitudinal, thick tracts ( Fig. 3C ). In the deeper region of the choanosome there are more spicules strewn in confusion, especially oxeas II ( Fig. 3C ). Dark brown, almost black, pigmented cells are common. Spicules ( Fig. 4 ) (average of all specimens; individual measurements in Table 1 ). Two categories of oxeas were observed: (1) Oxeas I, smooth, stout, slightly curved, usually with conical ends, but acerate, blunt, mucronate, strongylote and stylote ends are common ( Figs. 4A–B ). Their shape is rather uniform, but the spicules have a large size range (160–226–281 / 6–12–23 μm). (2) Oxeas II, thin, smooth, slightly curved or double bent, with acerate ends (111–177–245 / 1–3–9 μm) ( Fig. 4C ). The oxeas II could be developmental stages. Microscleres are very thin toxas, with a central swelling, variable in shape, some with low and straight axis, similar to raphides, and others with high and curved axis: 21–46–77 μm ( Fig. 4 D’–D’). These toxas were observed only in nine of the 22 specimens examined. Bathimetric distribution and ecology. Neopetrosia carbonaria is an intertidal and shallow water species, found between 0.5–3 m depth. It is often found in reef environments, on the under-surfaces of dead corals or buried in muddy sediment. The specimens from Northeastern Brazil has been observed growing among the bryozoans of the genus Stylopoma Levinsen, 1909 , the zoanthid Palythoa caribaeorum Duchassaing & Michelotti, 1860 , other sponges and calcareous algae. Distribution. U.S. Virgin Islands (Duchassaing & Michelotti 1864), Florida ( Van Soest 1980 ), Gulf of Mexico (Ŗtzler et al. 2009), Curaçao ( Van Soest 1980 ; Arndt 1927 ), Jamaica ( Hechtel 1965 ; Pulitzer-Finali 1986 ), Barbados ( Hechtel 1969 ), Dry Tortugas ( de Laubenfels 1936 ), Puerto Rico ( Van Soest 1980 ; Pulitzer-Finali 1986 ), Colombia ( Zea 1987 ; Zea et al. 2014 ), Bocas del Toro ( Díaz 2005 ), Belize (Ŗtzler et al. 2000), Martinique ( Pérez et al. 2017 ). Northeastern Brazil : Ceará , Rio Grande do Norte , Paraíba , Pernambuco , Alagoas and Bahia States ( Hechtel 1976 ; present study). TABLE 1. Comparative data of spicules dimensions, locality and color among examined specimens of Neopetrosia carbonaria . Values are in micrometres (μm), expressed as follows: minimum–average–maximum, length/width, n = 30.

Specimens examined	Locality	Color	Oxea I	Oxea II	Toxas
MNHN-LBIM-DT 545 (Holotype)	American Seas	Black	182–215–238/6– 10–13	115–181–205/2– 4–5	–
UFPEPOR 1637	Paraíba, NE Brazil	Dark green	205–233– 245/10–15–20	150–187–210/2– 4–9	45–52–63 (n=11)
UFPEPOR 1853	Paraíba, NE Brazil	Reddish brown	202–221–245/7– 12–15	155–190–228/1– 3–5	–
UFPEPOR 1854	Paraíba, NE Brazil	Reddish brown	172–217– 248/10–13–16	152–182–198/1– 3–5	–
UFPEPOR 1855	Paraíba, NE Brazil	Reddish brown	200–212–240/6– 12–15	150–185–213/1– 3–5	–
UFPEPOR 1458	Pernambuco, NE Brazil	Dark green	207–225– 243/12–14–15	130–163–245/1– 2–4	–
UFPEPOR 1472	Pernambuco, NE Brazil	Dark green	212–228–240/7– 13–15	137–172–193/1– 2–4	48 (n = 1)
YPM IZ 8960	Pernambuco, NE Brazil	–	247–262– 281/12–17–22	134–184–213/2– 3–6	46–61–77 (n=12)
MNRJ 17400	Ceará, NE Brazil	–	189–218– 242/10–11–15	130–167–201/1– 2–2	34–44–60 (n=18)
MNRJ 17741	Ceará, NE Brazil	Dark green	213–234– 261/10–12–15	122–165–199/1– 1–3	–
MNRJ 17766	Ceará, NE Brazil	Brown	203–234– 266/10–13–15	148–186–218/1– 2–3	–
MNRJ 17777	Ceará, NE Brazil	Brown	203–219– 237/10–11–15	119–161–208/1– 2–3	–
MNRJ 17782	Ceará, NE Brazil	Dark green	189–221– 242/10–10–15	124–170–205/1– 2–4	–
MNRJ 17783	Ceará, NE Brazil	Reddish brown	174–214– 232/10–10–15	111–177–209/1– 2–3	–
MNRJ 17832	Ceará, NE Brazil	Dark green	203–223– 237/10–10–15	140–177–210/1– 2–4	27–36–42 (n=7)
MNRJ 17840	Ceará, NE Brazil	Dark green	203–227– 252/10–12–15	123–171–215/1– 2–3	28–35–45 (n=7)
MNRJ 17866	Ceará, NE Brazil	Brown	213–232– 266/10–12–15	137–180–211/1– 2–3	21–33–41 (n=30)
MNRJ 17887	Ceará, NE Brazil	Dark green	160–211– 257/10–11–15	132–165–206/1– 2–4	–
MNRJ 18033	Rio Grande do Norte, NE Brazil	Dark green	194–217– 252/10–10–15	130–170–212/1– 2–3	26–35–42 (n=30)
MNRJ 18690	Abrolhos Bank, Bahia, NE Brazil	Dark green, almost black	203–222– 242/10–10–15	141–178–211/1– 2–3	–
MNRJ 4713	Alagoas, NE Brazil	Dark green, almost black	189–220– 257/10–16–19	131–169–201/1– 2–3	29–59–70 (n=11)
MNRJ 17035	Alagoas, NE Brazil	Dark green	218–235– 257/10–16–19	125–175–206/1– 3–5	–
MNRJ 9405	Panama	Black	237–254– 273/11–16–23	190–212–233/1– 3–8	50–59–70 (n=9)

FIGURE 2. Underwater photographs of Neopetrosia carbonaria specimens. (A) UFPEPOR 1853, (B) UFPEPOR 1854, (C) UFPEPOR 1637, (D) MNRJ 17035, (E) MNRJ 17783, (F) UFPEPOR 1855, (G) MNRJ 17766, (H) MNRJ 17782. FIGURE 3. Anatomical characters of Neopetrosia carbonaria . ( A–B) Tangential view of the ectosomal skeleton, (C) Cross section of the skeleton, showing the subdermal layer and the inner choanosome. Scale bars: (A–B) 500 μm, (C) 1 mm. Taxonomic remarks. Neopetrosia carbonaria is defined here by the thick encrusting to repent or ramose growing form, with reddish brown to brown color and dark green to black color in vivo , two categories of oxeas as megascleres and rare raphidiform toxas as microscleres. Despite the presence of oxeas larger than 200 μm, which approach this species to the genus Xestospongia , the studied specimens were assigned to Neopetrosia based on its skeletal arrangement that is somewhat irregular, with poorly defined multispicular tracts, but with a well-developed subdermal layer. Some studied specimens showed similar features with the holotype of Neopetrosia carbonaria , such as the thickly encrusting shape, with oscular projections, the very dark color, the presence of a subdermal layer and pigmented cells in the skeleton, and stout and fusiform oxeas, with similar sizes (Topsent 1930). However, new morphological features and spicules types were revealed in some Brazilian populations of N. carbonaria , such as the presence of raphidiform toxas as microscleres and a wide range of color, including reddish brown, brown and dark green color. We found toxas in nine of the 21 studied specimens from the Northeastern Brazil , in some pictures of spicule slides of Neopetrosia carbonaria sensu Hechtel, 1976 (as Adocia carbonaria , YPM IZ 8960) from Pernambuco State , Northeastern Brazil , and in one specimen from Bocas del Toro , Panama (MNRJ 9405) ( Tab. 1 ). These microscleres can be quite rare and occur in both black and brown sponges. Despite the absence of toxas in the fragment of N. carbonaria holotype reexamined here (MNRJ 21834, Tab. 1 ), these microscleres were reported in some populations of N. carbonaria from the Caribbean coast of Colombia , as toxiform raphides ( Zea 1987 ; Zea et al. 2014 ). Due the absence and rarity of raphidiform toxas in some specimens analyzed, unrelated to its color, we agree with Zea (1987) and assume that these differences are intraspecific. However, all the records of Neopetrosia carbonaria need a revaluation, since the presence of toxas among microscleres can be easily overlooked. In the field, reddish brown and brown specimens of N. carbonaria could be confused with N. subtriangularis and Calyx podatypa ( De Laubenfels, 1934 ) . Neopetrosia subtriangularis can be distinguished from N. carbonaria by its smaller and thinner oxeas (104–120–144 / 2–3–4 μm versus 160–226–281 / 6–12–23 μm in N. carbonaria ) and C. podatypa has a more regular choanosomal skeleton, with well-defined uni- to paucispicular tracts, forming triangular to polygonal meshes, without a subdermal layer ( Zea et al. 2014 ). In addition, none of these species has toxas and the dark green to black color of N. carbonaria .