‘Endless forms most beautiful’: taxonomic revision of the planarian Geoplana vaginuloides (Darwin, 1844) and discovery of numerous congeners (Platyhelminthes: Tricladida)

‘Endless forms most beautiful’: taxonomic revision of the planarian Geoplana vaginuloides... Abstract In 1832, Darwin discovered two ‘elegantly coloured true Planaria’ in Rio de Janeiro, one of which he later formally named as Planaria vaginuloides (Darwin, 1844). Currently named Geoplana vaginuloides (Darwin, 1844), it is the type species of the genus and of the subfamily Geoplaninae. Many chromatic patterns of the body were recorded for the species, but its polyphyletic status was recently pointed out. Herein we revise the three-species-rich genus by combining morphological and molecular data within an integrative taxonomic framework. Our results support the monophyly of the genus, but also reveal that there are at least four nominal species recorded in the literature under the name of G. vaginuloides. This number increases to ten if the diagnostic features used by former authors are applied, i.e. the eye-catching dorsal colour pattern and the typical, extraordinarily long penis papilla. Our results also indicate that Geoplana chita Froehlich, 1956 is a complex of species and that Geoplana pulchella Schultze & Müller, 1857 is monophyletic. We describe herein 13 species of the genus. Surprisingly, the number of species of Geoplana was overshadowed by their beauty. Atlantic forest, Continenticola, DNA, genes, Geoplaninae, histology, microcomputed tomography, morphology, phylogeny, systematics INTRODUCTION Land planarians are chiefly pantropical in distribution, and the Neotropical region houses ~260 species, all considered as members of Geoplaninae Stimpson, 1857. Throughout its distribution, the Atlantic forest is credited to be one of the areas with the highest species densities worldwide (Sluys, 1998, 2016). It was in this region that a few specimens of land planarians caught the attention of Charles Darwin in the 19th century. Enchanted by the tropical forest near Rio de Janeiro, southeastern Brazil, Darwin confessed that ‘nothing has so much interested me as finding two species of elegantly coloured true Planaria inhabiting the dewy forest!’ (1832, letter to H. S. Henslow). Land planarians were not phytovorous, as Darwin thought (1832, p. 48, in Keynes, 2000); they are predators of soil invertebrates (Froehlich, 1966). Back in England, Darwin described a few new species of land flatworms from around the world, including the colourful Planaria vaginuloides Darwin, 1844 (currently in Geoplana Schultze & Müller, 1857), which he had found in the surroundings of Rio de Janeiro. Darwin lived in the pre-histological era; therefore, the formal description of the species was based merely on the external aspect. With respect to its chromatic pattern, he described G. vaginuloides as having “Sides and foot coloured dirty ‘orpiment orange’; above, with two stripes on each side of pale ‘primrose-yellow’, edged externally with black; on centre of the back a stripe of glossy black; these stripes become narrow towards both extremities”. The information provided in the original description is all we have available to diagnose G. vaginuloides, because the type material seems to be lost (H. D. Jones, personal communication). Riester (1938) and Froehlich (1956a) examined additional worms from Teresópolis [State of Rio de Janeiro (RJ)], Marcus (1951, 1952) from São Paulo [State of São Paulo (SP)], and Froehlich (1958) from Itanhaém (SP). Additionally, Froehlich (1955a) examined one specimen from an unknown locality (see Supporting Information, Table S1). Based on these studies, up to seven chromatic body patterns are currently known for this species (Darwin, 1844; Riester, 1938; Marcus, 1952; Froehlich, 1958). Riester (1938) and Marcus (1951) examined the internal organs from histological slides, and Froehlich (1958) summarily studied these organs in one cleared specimen. Apart from the records of G. vaginuloides mentioned above, Prudhoe (1949) reported this species from Trinidad and Tobago. However, Marcus (1951, 1952) doubted the conspecificity of Prudhoe’s specimens owing to the very short copulatory apparatus relative to its body length (see also Froehlich, 1967; Carbayo & Leal-Zanchet, 2001). Kawakatsu et al. (2002) considered the specimens from Trinidad and Tobago to represent a new species of Geoplana, viz., Geoplana prudhoei Kawakatsu, Ogren, Froehlich & Sasaki, 2002. No more taxonomic works have been published on G. vaginuloides, with the exception of a molecular phylogenetic study of the subfamily Geoplaninae, in which the polyphyletic status of G. vaginuloides (represented by two individuals) was pointed out in a recent paper (Carbayo et al., 2013). In that paper, the diagnosis of Geoplana was also emended, and 100 species of the genus were transferred to other genera or were considered incertae sedis. Currently, there are only three species in the genus, namely, G. vaginuloides, Geoplana pulchella Schultze & Müller, 1857 and Geoplana chita Froehlich, 1956, all from Brazil. The only taxonomic study of G. chita is the original description, which included details of its internal organs (Froehlich, 1956b). The original description of G. pulchella, from Blumenau [State of Santa Catarina (SC), Brazil], is based on its external aspect (Schultze & Müller, 1857). An additional specimen from Brusque (state of SC, Brazil) was examined by du Bois-Reymond Marcus (1951). However, this specimen was considered not to be conspecific and is currently placed in Obama Carbayo et al., 2013. Froehlich (1955b) reported an immature specimen of G. pulchella from the type locality. The polyphyletic status of G. vaginuloides is problematic owing to the fact that it is the type species of the genus and of the subfamily (Froehlich, 1955a). Consequently, the nomenclatural stability of the genus and subfamily is dependent on the unequivocal circumscription of this taxon. This work is aimed at an integrative taxonomic revision of the genus. We analysed the morphology of the material originally examined by Riester (1938), Marcus (1951, 1952) and Froehlich (1956a), and additional specimens collected by or donated to the late Professor Eudóxia Maria Froehlich (University of São Paulo). In addition, we have included specimens collected by ourselves or donated to us, from which we obtained mitochondrial and nuclear gene fragments to infer their phylogenetic relationships. Finally, we have also revised the taxonomy of the two other nominal species of the genus (see Carbayo et al., 2013), i.e. G. chita and G. pulchella Schultze & Müller, 1857, by examining the available type specimens and additional individuals collected by us and from Eudóxia’s collection, and we describe, for the first time, the internal morphology of G. pulchella. MATERIAL AND METHODS Fieldwork We actively collected planarians in areas covered with Atlantic Forest in the States of Rio de Janeiro, São Paulo, Paraná and Santa Catarina, Brazil (Fig. 1). After collection, they were photographed and their external aspect was recorded before they were killed in boiling water. Before fixing them for histological studies, we removed a small piece of the worm and preserved it in absolute ethanol and stored this material in a freezer. Specimens from E. M. Froehlich’s collection were probably fixed with Susa. Specimens collected in 2005 or later were fixed in 100% ethanol or in 10% formalin buffered solution. Figure 1. View largeDownload slide Distribution map of the species of Geoplana addressed in this paper. Abbreviations for the Brazilian States are as follows: PR, Paraná; RJ, Rio de Janeiro; RS, Rio Grande do Sul; SC, Santa Catarina; SP, São Paulo. Figure 1. View largeDownload slide Distribution map of the species of Geoplana addressed in this paper. Abbreviations for the Brazilian States are as follows: PR, Paraná; RJ, Rio de Janeiro; RS, Rio Grande do Sul; SC, Santa Catarina; SP, São Paulo. Histology We cut the specimens into several pieces, dehydrated these in a graded ethanol series, cleared them in clove oil, and infiltrated and embedded them in Paraplast tissue embedding medium. We then sectioned these tissue blocks at 3–7 μm intervals using a retracting rotary microtome, affixed the sections with albumin and glycerol (1:1) onto glass slides placed on a hotplate, and stained them following Cason (1950). Finally, sections were dehydrated in a graded ethanol series, cleared in xylene and mounted in synthetic balsam (Merck and Synth). We examined the slides with an optical compound microscope and made drawings of the pharynges and the copulatory apparatuses with the aid of a camera lucida attached to the microscope. Photomicrographs were taken with a DP72 digital camera attached to the microscope, and images were edited in GIMP (GNU Image Manipulation Program 2.8.16; The GIMP team www.gimp.org, 1995–2016). Colour descriptions of the body of live or fixed specimens follow online RAL palette colours (© RAL gemeinnützige GmbH, available at https://www.ral-farben.de/uebersicht-ral-classic-farben.html?&L=1) through comparison with digital images of the specimens on a computer screen. The thickness of cutaneous musculature is expressed as a percentage of body height in the prepharyngeal region (CMI). The figures of dorsal, sagittal and horizontal views are oriented so that the anterior end of the body is to the left. The material we collected or that was donated to us was deposited at the Museu de Zoologia da Universidade de São Paulo (MZUSP). We also examined material deposited in the Senckenberg Museum Frankfurt (SMF) and in Professor Eudóxia Maria Froehlich’s Collection (EMF, provisionally under the care of F.C.; see details in the respective section ‘Material examined’ for each species). Microcomputed tomography Virtual two-dimensional sections were made of specimens MZUSP PL 2080 and MZUSP PL 2095. The specimens were submerged in 100% ethanol containing 1% iodine in absolute ethanol for approximately for 4 weeks. One day before scanning, the specimens were rinsed in absolute ethanol for 5 min and, subsequently, the region of the copulatory apparatus was scanned in an X-ray micro-computed tomography (µCT) SkyScan1272 scanner or Skyscan 1272 (Bruker MicroCT, Kontich, Belgium). The operational settings for scanning the specimen MZUSP PL 2080 were as follows: 21 kV, source current of 175 μA, image pixel size of 3.249953 μm, no filter applied, depth of 16 bits, exposure of 1458 ms, rotation step of 0.080°, and frame averaging of 2. The operational settings for scanning the specimen MZUSP PL 2095 were as follows: 73 kV, source current of 104 μA, image pixel size of 5.69 μm, no filter applied, depth of 16 bits, exposure of 1000 ms, rotation step of 0.15°, and frame averaging of 12. Reconstructions were carried out as bmp formatted images using the software Nrecon version 1.6.6.0 (Bruker MicroCT) with the following settings: smoothing = 0, ring artefact correction = 15, beam hardening correction = 63% (specimen MZUSP PL 2080) and 0% (MZUSP PL 2095), and HU calibration = off. Sets of virtual sections were reconstructed with the software package DataViewer (http://www.skyscan.be/products/downloads.htm), and a selection of sets with the original grey scales were saved. The contrast of grey-scale virtual sections was enhanced with GIMP (GNU Image Manipulation Program 2.8.10). After the virtual sections had been acquired, we rinsed the specimens in 80% ethanol and, subsequently, dehydrated and treated them, following the histological procedure described in the previous subsection. Material examined Specimens identified and described are listed below in the ‘Material examined’ section for the respective species. We made an effort to find and identify all other specimens of the genus recorded in the literature. Those specimens whose identification could not be ascertained are listed in the Supporting Information (Table S1). Those specimens are immature, the histological preparations of their copulatory apparatus are of limited quality, or they have been lost. In the Supporting Information (Table S1) there are also recently collected representatives of undescribed species of Geoplana (named Geoplana sp. 1–4 in the molecular analyses) that are outside the scope of the present work. Acquisition of molecular data We selected 42 specimens of putative Geoplana members for molecular studies. Genomic DNA extractions were performed with the Wizard Genomic DNA Purification kit (Promega, Madison, WI, USA) following Álvarez-Presas et al. (2011). Polymerase chain reaction (PCR) was used to amplify partial sequences of the mitochondrial region of Cytochrome Oxidase I (COI) and the nuclear region of the Elongation Factor-1-alpha promoter (EF-1-α). Amplifications were performed in a 20 μL volume containing 1 μL of DNA, 4 μL of 5× Phusion Buffer GC (Thermo Scientific) 1.5 mM Tris–HCl (pH 8.4), 50 mM KCl, 1 μL of l M dNTPs, 1.0–3.0 mM MgCl2, 1 μL of l M of each primer, and 0.2 μL of Phusion High-Fidelity DNA Polymerase (Thermo Scientific). General PCR conditions included initial denaturation for 3 min at 98 °C, 35 cycles of denaturation for 10 s at 98 °C, annealing for 30 s at specific temperatures (see below), extension for 1 min at 72 °C, and a final extension for 5 min at 72 °C. Amplifications and sequencing were performed with following primer sets: COI with ‘FlatwormCOI-F’ (5′-GCAGTTTTTGGTTTTTTGGACATCC-3′) and ‘FlatwormCOI-R’ (5′-GAGCAACAACATAATAAGTATCATG-3′) (Sunnucks et al., 2006) at 45 °C; BarT 5′-ATGACDGCSCATGGTTTA-3′, BarS 5′-GTTATGC CTGTAATGATTG-3′ (Álvarez-Presas et al., 2011) and COIR 5′-CCWGTYARMCCHCCWAYAGTAAAA-3′ (Lázaro et al., 2009) at 45 °C; and EF-1-α with EFplatF 5′-GATTGCYCCWGGYCATCG-3′ and EFplatR 5′-GCRATWGAYTCGTGRTGC-3′ (Carbayo et al., 2013) at 45–48 °C. The PCR products were purified using an Agencourt AMPure XP DNA Purification and Cleanup kit (Beckman Coulter Inc.). Products were subsequently cycle sequenced directly from both forward and reverse directions using ABI Big-Dye Sequence Terminator (v. 3.1), cleaned with ethanol precipitation, and sequenced on an ABI Prism Genetic Analyzer (3131XL) automated sequencer (Applied Biosystems/ThermoFisher). Sequence reads were assembled using the Consed/Phred/Phrap package (Ewing & Green 1998; Ewing et al., 1998; Gordon, Abajian & Green, 1998; Gordon, Desmarais & Green, 2001). Before submitting the data to phylogenetic analysis, we aligned the sequences with MAFFT (Katoh et al., 2002) using the G-INS-i iterative refinement method with 1000 cycles, and visualized and edited the sequences in BioEdit (Hall, 1999). Aligned sequences of COI and EF-1-α were checked for stop codons using the DNA-to-protein translation online resource by Bikandi et al. (2004). Phylogenetic inference We considered two datasets: the first one, hereafter referred as all-terminals, included all relevant terminals for which we had sequences of COI and/or EF-1-α, which included terminals with missing fragments; the second, hereafter referred as selected-terminals, included a subset of the all-terminals dataset, which included terminals for which both fragments (i.e. COI and EF-1-α) were available. These two datasets allowed us to evaluate clade stability owing to missing data, which was relevant to us because for some putative species we had only one of the fragments. We proceeded with phylogenetic inferences by using two optimality criteria: maximum parsimony (MP) and maximum likelihood (ML). Phylogenetic analyses under parsimony were carried out with TNT (Goloboff, Farris & Nixon, 2008) using its New Technology searches (Goloboff, 1999; Nixon, 1999) with the following search parameters: rep 100, ratchet 20, fuse 20 and hold 10. For ML analyses, model selection was performed in jModelTest (version 2.1.6; Guindon et al., 2003; Darriba et al., 2012), considering 88 candidate models ranked by corrected Akaike information criterion (AICc) scores, and tree searches were performed in GARLI (version 2.0; Zwickl, 2006), considering both single and partition models for each genomic region for 1000 independent search replicates. Following Carbayo et al. (2013), we considered five representatives of Rhynchodeminae as outgroups: Arthurdendyus triangulatus (Dendy, 1895); Artioposthia testacea Hutton, 1880; Australoplana sp.; Dolichoplana striata Moseley, 1877; and Endeavouria septemlineata (Hyman, 1939) (see Table 1). Table 1. Individuals used in the study, museum code used in the figures and text, collecting locality data, identification base and GenBank accession numbers for all sequences Species Field number Specimen’s accession number (MZUSP PL) Collecting locality Collecting date Latitude Longitude Identification COI EF-1-alpha Rhynchodeminae Arthurdendyus triangulatus (Dendy, 1895) GenBank DQ666027 KC614559 Artioposthia testacea Hutton, 1880 GenBank AF178305 KC614560 Australoplana sp. GenBank DQ666028 KC614558 Dolichoplana striata Moseley, 1877 F0379 1005 Igrejinha, RS 10 July 2004 −29.57200 −50.77800 GenBank KC608226 KC614465 Endeavouria septemlineata (Hyman, 1939) F0988 657 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608233 KC614471 F0192 1184 Maquiné, RS 10 February 2004 −29.67774 −50.20640 GenBank KC608222 KC614461 Geoplaninae Cephaloflexa araucariana Carbayo & Leal-Zanchet, 2003 F3387 1073 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608316 KC614536 F3426 1076 Floresta Nacional de São Francisco de Paula, RS 23 January 2009 −29.42739 −50.39072 GenBank KC608319 KC614539 Cephaloflexa bergi (Graff, 1899) F1034 303 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608238 KC614472 F1038 305 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608240 KC614474 Cephaloflexa sp. 1 F2103 1211 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608272 – F2108 1019 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608274 KC614501 Choeradoplana banga Carbayo & E. M. Froehlich, 2012 F2023 1000 Parque Estadual Serra da Cantareira, SP 30 January 2008 −23.42916 −46.63250 GenBank KC608267 – F3011 1002 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608301 KC614523 Choeradoplana bocaina Carbayo & E. M. Froehlich, 2012 F2104 999 Parque Nacional Serra da Bocaina, SP 10 February 2008 −22.75222 −44.62194 GenBank KC608273 KC614500 F2803 998 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 GenBank KC608283 KC614509 F2822 997 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 GenBank KC608288 – Choeradoplana gladismariae Carbayo & E. M. Froehlich, 2012 F3092 1003 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608306 KC614528 F3802 1004 Parque Estadual Intervales, SP 7 July 2009 −24.27638 −48.41556 GenBank KC608326 – Choeradoplana iheringi Graff, 1899 F3449 533 Floresta Nacional de São Francisco de Paula, RS 2 April 2012 −29.42367 −50.38711 GenBank KC608320 KC614540 Choeradoplana pucupucu Carbayo et al., 2017 F2840 540 Parque Nacional Serra da Bocaina, SP September 2008 −22.74277 −44.61639 GenBank KC608293 KC614517 Cratera crioula (E. M. Froehlich, 1955) F3708 1078 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608323 KC614543 F3715 1079 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608324 KC614544 Cratera pseudovaginuloides (Riester, 1938) F1244 670 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608251 KC614482 F1245 671 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608252 KC614483 Cratera tamoia (E. M. Froehlich, 1955) F1139 665 Parque Nacional da Serra dos Órgãos, RJ 15 January 2007 −22.45528 −42.99750 GenBank KC608246 KC614478 F1336 672 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608254 KC614484 Geobia subterranea (Schultze & Müller, 1857) F0358 650 Colégio Cristo Rei, São Leopoldo, RS 30 May 2004 −29.78828 −51.15084 GenBank KC608225 KC614464 F1355 673 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608255 KC614485 Geoplana apua Almeida & Carbayo sp. nov. F5236 2072 Nova Iguaçú, RJ 4–6 April 2012 Holotype MG653226* – Geoplana boraceia Almeida & Carbayo sp. nov. F4112 2070 Estação Biológica de Boraceia, SP 21 December 2009 −23.65389 −45.88917 Holotype – MG653220* F4358 1086 São Sebastião, SP 26 February 2010 −23.75203 −45.631027 Paratype KC608329 KC614547 F4389 2123 São Sebastião, SP 26 February 2010 −23.746 −45.632 Paratype MG653225* – F4409 1087 São Sebastião, SP 27 February 2010 −23.75203 −45.631027 External morphology similar to type material KC608330 KC614548 F6026 2095 São Sebastião, SP 11 January 2014 −23.74439 −45.63715 Paratype MG653227* MG653194* F6055 2125 São Sebastião, SP 11 January 2014 Paratype MG653237* MG653205* F6068 2124 São Sebastião, SP 12 January 2014 Paratype MG653244* – F6357 2073 São Sebastião, SP 27 September 2014 −23.74685 −45.63213 Paratype MG653238* MG653206* F6575 2077 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653239* MG653208* F6578 2078 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653240* MG653209* Geoplana cambara Almeida & Carbayo sp. nov. F1614 1009 Parque Nacional Saint Hilaire / Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank (named G. vaginuloides in GenBank) KC608262 KC614492 F1635 2063 Parque Nacional Saint Hilaire/Lange, PR 11 January 2008 Paratype MG653229* MG653198* Geoplana cananeia Almeida & Carbayo sp. nov. F4096 2067 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Holotype MG653223* MG653196* F4097 2068 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653233* MG653201* F4098 2069 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653224* – Geoplana caraguatatuba Almeida & Carbayo sp. nov. F6727 2080 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 May 2016 −23.594012 −45.429672 Paratype MG653247* – F6976 2126 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 January 2016 −23.593880 −45.429746 Paratype MG653251* – Geoplana chita Froehlich, 1956 F1710 715 Morretes, PR 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653218* F1730 716 Morretes, PR-25.469604 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653219* Geoplana ibiuna Almeida & Carbayo sp. nov. F3562 2066 Parque Estadual do Jurupará/Ibiúna, SP February 2009 Holotype MG653232* – F6907 2081 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653248* MG653214* F6908 2082 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653249* MG653215* F6909 2083 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653250* – Geoplana iporanga Almeida & Carbayo sp. nov. F3166 2065 Parque Estadual de Intervales, SP 13 December 2008 −24.269387 −48.405467 Holotype MG653231* MG653200* F3132 2064 Parque Estadual de Intervales/Ribeirão Grande, SP 13 December 2008 −24.269387 −48.405467 Paratype – MG653221* Geoplana mogi Almeida & Carbayo sp. nov. F1022 2061 Mogi das Cruzes, SP 27 May 2006 −23.451536 −46.227993 Paratype – MG653217* Geoplana paranapiacaba Almeida & Carbayo sp. nov. F6660 2079 Cidade Turística de Paranapiacaba, SP 5 June 2015 −23.77746 −46.30145 Holotype MG653246* MG653213* Geoplana piratininga Almeida & Carbayo sp. nov. F1163 666 Parque Jardim Previdência/SP 17 June 2007 −23.579662 −46.727503 Paratype KC608247 KC614479 F0509 2060 Parque Ecológico do Tietê/SP 27 May 2005 −23.490918 −46.516424 Paratype MG653222* MG653195* Geoplana pulchella Schultze & Müller, 1857 F3249 1068 Parque Estadual da Serra do Tabuleiro, SC 14 January 2009 −27.98139 −48.74806 External aspect similar to that of original description KC608310 KC614532 F6528 2075 RPPN Vale das Pedras, SC 16 January 2015 −27.61695 −049.34778 External aspect similar to that of original description MG653245* MG653212* F6544 2075 RPPN Vale das Pedras, SC 18 January 2015 −27.61696 −049.34813 External aspect similar to that of original description – MG653207* Geoplana quagga Marcus, 1951 F1003 1188 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 Genbank KC608236 – Geoplana sp. 1 F2934 2127 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653241* – F2943 2128 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653242* MG653210* F2944 2129 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653243* MG653211* Geoplana sp. 2 F5145 2130 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653234* MG653202* F5163 2131 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653235* MG653203* F5204 2132 Parque Nacional de Itatiaia, RJ 6 April 2012 Molecular and morphological support MG653236* MG653204* Geoplana sp. 3 F2638 2133 Parque Estadual de Intervales, SP 24 December 2008 −24.269387 −48.405467 Molecular support MG653230* MG653199* Geoplana sp. 4 F1707 2134 Morretes, PR 13 January 2008 −25.469604 −48.813739 Molecular support MG653252* MG653216* Geoplana vaginuloides (Darwin, 1844) F6387 2074 Parque Estadual da Pedra Branca, RJ 13 December 2014 −22.93707 −43.46098 Neotype MG653228* MG653197* Geoplana goetschi Riester, 1938, incertae sedis F2870 412 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608294 – Geoplana hina Marcus, 1951, (incertae sedis) F1613 1008 Parque Nacional Saint Hilaire/Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank KC608261 KC614491 Geoplana phocaica Marcus, 1951 (incertae sedis) F2827 457 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608289 KC614514 Geoplana phocaica Marcus, 1951 (incertae sedis) F2830 460 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608290 KC614515 Geoplaninae 1 F2033 1193 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.75000 −44.61666 GenBank KC608269 – Geoplaninae 2 F2807 1050 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 Named as Geoplana sp. 1 in GenBank KC608284 KC614510 F2821 1052 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 Named as Geoplana sp. 1 in GenBank KC608287 KC614513 Geoplaninae 3 F2783 1048 Parque Estadual Intervales, SP 29 July 2008 −24.27333 −48.41694 Named as Geoplana sp. 2 in GenBank KC608281 KC614508 Geoplaninae 4 F2796 1194 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 3 in GenBank KC608282 – Geoplaninae 5 F2031 1014 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.73333 −44.60000 Named as Geoplana sp. 4 in GenBank KC608268 KC614497 F2809 1051 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 4 in GenBank KC608285 KC614511 Geoplaninae 6 F2189 348 Parque Estadual do Desengano, RJ 18 March 2008 −21.87667 −41.91972 Named as Geoplana sp. 5 in GenBank KC608278 KC614505 Geoplaninae 7 F3191 1066 Parque Estadual da Serra do Tabuleiro, SC 9 January 2009 −27.95989 −48.75761 Named as Geoplana sp. 6 in GenBank KC608308 KC614530 F3247 1067 Parque Estadual da Serra do Tabuleiro, SC 13 January 2009 −27.98139 −48.74806 Named as Geoplana sp. 6 in GenBank KC608309 KC614531 Gusana sp. 1 F4421 1088 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608331 KC614549 F4428 1089 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608332 – Imbira guaiana (Leal-Zanchet & Carbayo, 2001) F0432 653 Floresta Nacional de São Francisco de Paula, RS 1 August 2004 −29.41666 −50.38333 GenBank KC608229 KC614467 F3361 1071 Floresta Nacional de São Francisco de Paula, RS 20 January 2009 −29.42367 −50.38711 GenBank KC608314 – Imbira marcusi Carbayo et al., 2013 F1041 1190 São Sebastião, SP 8 September 2006 −23.75203 −45.631027 GenBank KC608241 – F2833 404 Parque Nacional Serra da Bocaina, SP September 2008 −22.75222 −44.62194 GenBank KC608291 KC614516 Issoca jandaia Froehlich, 1955 F2043 1015 Parque Nacional Serra da Bocaina, SP 9 February 2007 −22.78222 −44.60583 GenBank KC608270 KC614498 Issoca rezendei (Schirch, 1929) F1182 667 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608248 KC614480 F1679 1010 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608263 KC614493 Issoca sp. 1 F2158 1020 Parque Estadual do Desengano, RJ 16 March 2008 −21.87694 −41.92583 GenBank KC608275 KC614502 F2507 1045 Reserva Biológica Augusto Ruschi, ES 24 May 2008 −19.88740 −40.54319 GenBank KC608279 KC614506 F4085 1085 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608328 KC614546 Luteostriata abundans (Graff, 1899) no voucher São Leopoldo, RS 1998 −29.78828 −51.15084 GenBank KC631621 KC614556 F0205 646 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608223 KC614462 F0238 648 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608224 KC614463 Luteostriata ceciliae (Froehlich & Leal-Zanchet, 2003) F3451 1077 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608321 KC614541 Luteostriata ernesti (Leal-Zanchet & Froehlich, 2006) no voucher São Paulo, SP 2005 −23.49080 −46.52090 GenBank – KC614555 F3358 1070 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608313 KC614534 Luteostriata graffi (Leal-Zanchet & Froehlich, 2006) F0384 652 Taquara, RS 28 January 2010 −29.660524 −50.746782 GenBank KC608227 KC614466 F0440 654 Morro das Aranhas, Florianópolis, SC 9 September 2004 −27.468485 −48.377724 GenBank KC608230 KC614468 F3381 1072 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42963 −50.39780 GenBank KC608315 KC614535 Luteostriata muelleri (Diesing, 1861) No voucher GenBank – KC614554 F3268 1199 Parque Estadual da Serra do Tabuleiro, SC 15 January 2009 −27.84278 −48.92583 GenBank KC608311 – Luteostriata sp. F1192 668 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608249 KC614481 Luteostriata sp. 1 F1036 659 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608239 KC614473 F1078 663 Estação Biológica de Boraceia, Salesópolis, SP, Brazil 2 February 2006 −23.65388 −45.88910 GenBank KC608243 KC614476 Luteostriata sp. 2 F0441 655 Santinhos, Florianópolis, SC 9 September 2004 −27.46370 −48.38100 GenBank KC608231 KC614469 Matuxia matuta (E. M. Froehlich, 1955) F2184 1021 Parque Estadual do Desengano, RJ 17 March 2008 −27.95989 −48.75761 GenBank KC608276 KC614503 F2187 1022 Parque Estadual do Desengano, RJ 17 March 2008 −21.87138 −41.91472 GenBank KC608277 KC614504 Matuxia tuxaua E. M. Froehlich, 1955 F1275 1192 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608253 – F3059 1058 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608302 KC614525 Notogynaphallia albonigra (Riester, 1938) F4081 1083 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608327 KC614545 Notogynaphallia parca (E. M. Froehlich, 1955) F2972 1196 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.42914 −46.63250 GenBank KC608298 – Notogynaphallia sexstriata (Graff, 1899) F0792 656 Cidade Universitária, USP, São Paulo, SP 19 December 2005 −23.56671 −46.73017 GenBank KC608232 KC614470 F1413 680 Parque Nacional da Serra de Itajaí, SC 4 January 2008 −27.04836 −49.09200 GenBank KC608257 KC614487 Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015 F1399 1007 Parque Nacional da Serra do Itajaí, Blumenau, SC 4 January 2008 −27.05306 −49.08528 GenBank KC608256 KC614486 F1504 698 Parque Nacional da Serra do Itajaí, Blumenau, SC 6 January 2008 −27.05306 −49.08528 GenBank KC608259 KC614489 F1566 707 Parque Nacional da Serra do Itajaí, Blumenau, SC 7 January 2008 −27.05306 −49.08528 GenBank KC608260 KC614490 Obama burmeisteri (Schultze & Müller, 1857) F0989 658 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608234 – F2886 733 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608297 KC614520 Obama carinata (Riester, 1938) F3084 1062 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608304 KC614527 F3093 1064 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608307 KC614529 Obama josefi (Carbayo & Leal-Zanchet, 2001) F3389 1074 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608317 KC614537 F3403 1075 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608318 KC614538 Obama ladislavii (Graff, 1899) F1418 681 Parque Nacional da Serra de Itajaí, SC 5 January 2008 −27.04836 −49.09200 GenBank KC608258 KC614488 Paraba franciscana (Leal-Zanchet & Carbayo, 2001) F3357 1069 Floresta Nacional de São Francisco de Paula, RS 2 November 2006 −29.42894 −50.39200 GenBank KC608312 KC614533 F5485 935 Floresta Nacional de São Francisco de Paula, RS 12 August 1999 −29.36666 −50.38333 GenBank KC608335 KC614552 Paraba multicolor (Graff, 1899) F1736 594 Parque Nacional Saint Hilaire/Lange, PR 14 January 2008 −25.46972 −48.81222 GenBank KC608265 KC614495 F2997 1055 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608299 KC614521 Paraba rubidolineata (Baptista & Leal-Zanchet, 2005) F5482 936 Floresta Nacional de São Francisco de Paula, RS 26 August 1999 −29.36666 −50.38333 GenBank KC608333 KC614550 Pasipha chimbeva (E. M. Froehlich, 1955) F2813 403 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 GenBank KC608286 KC614512 F3010 476 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608300 KC614522 Pasipha pasipha (Marcus, 1951) F1841 1012 Parque Estadual Alberto Löfgren, São Paulo, SP 26 January 2008 −23.45925 −46.63730 GenBank KC608266 KC614496 F2871 1053 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608295 KC614518 Pasipha pinima (E. M. Froehlich, 1955) F1714 1011 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608264 KC614494 F2585 717 Parque Nacional da Serra dos Órgãos, RJ 10 July 2008 −22.45528 −42.99750 GenBank KC608280 KC614507 Pasipha rosea (E. M. Froehlich, 1955) F1061 660 Parque Estoril, São Bernardo do Campo, SP 22 October 2006 −23.77368 −46.51469 GenBank KC608242 KC614475 Pasipha sp. 2 F1138 664 Parque Nacional da Serra dos Órgãos, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608245 KC614477 Pasipha tapetilla (Marcus, 1951) F2878 732 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608296 KC614519 F5631B 938 São Paulo, SP May 2007 −29.36666 −50.38333 GenBank KC608336 KC614553 Polycladus sp. 1 F0397 1186 Valdívia (Chile) 2004 −39.81000 −73.23000 GenBank KC608228 – Supramontana irritata Carbayo & Leal-Zanchet, 2003 F3452 772 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42064 −50.39394 GenBank KC608322 KC614542 F5483 937 Floresta Nacional de São Francisco de Paula, RS 9 February 2006 −29.36666 −50.38333 GenBank KC608334 KC614551 Xerapoa hystrix Froehlich, 1955 F3057 1057 Parque Estadual Serra da Cantareira, SP 12 December 2008 −24.27638 −48.41556 GenBank – KC614524 Xerapoa pseudorhynchodemus (Riester, 1938) F0992 1187 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608235 – F1013 1189 Cidade Universitária, USP, São Paulo, SP 17 March 2006 −23.56671 −46.73017 GenBank KC608237 – Xerapoa trina (Marcus, 1951) F1134 1191 Cascata Imbuí, Teresópolis, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608244 – F1239 669 Parque Nacional da Serra dos Órgãos, RJ 15 December 2007 −22.45528 −42.99750 GenBank KC608250 – Species Field number Specimen’s accession number (MZUSP PL) Collecting locality Collecting date Latitude Longitude Identification COI EF-1-alpha Rhynchodeminae Arthurdendyus triangulatus (Dendy, 1895) GenBank DQ666027 KC614559 Artioposthia testacea Hutton, 1880 GenBank AF178305 KC614560 Australoplana sp. GenBank DQ666028 KC614558 Dolichoplana striata Moseley, 1877 F0379 1005 Igrejinha, RS 10 July 2004 −29.57200 −50.77800 GenBank KC608226 KC614465 Endeavouria septemlineata (Hyman, 1939) F0988 657 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608233 KC614471 F0192 1184 Maquiné, RS 10 February 2004 −29.67774 −50.20640 GenBank KC608222 KC614461 Geoplaninae Cephaloflexa araucariana Carbayo & Leal-Zanchet, 2003 F3387 1073 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608316 KC614536 F3426 1076 Floresta Nacional de São Francisco de Paula, RS 23 January 2009 −29.42739 −50.39072 GenBank KC608319 KC614539 Cephaloflexa bergi (Graff, 1899) F1034 303 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608238 KC614472 F1038 305 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608240 KC614474 Cephaloflexa sp. 1 F2103 1211 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608272 – F2108 1019 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608274 KC614501 Choeradoplana banga Carbayo & E. M. Froehlich, 2012 F2023 1000 Parque Estadual Serra da Cantareira, SP 30 January 2008 −23.42916 −46.63250 GenBank KC608267 – F3011 1002 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608301 KC614523 Choeradoplana bocaina Carbayo & E. M. Froehlich, 2012 F2104 999 Parque Nacional Serra da Bocaina, SP 10 February 2008 −22.75222 −44.62194 GenBank KC608273 KC614500 F2803 998 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 GenBank KC608283 KC614509 F2822 997 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 GenBank KC608288 – Choeradoplana gladismariae Carbayo & E. M. Froehlich, 2012 F3092 1003 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608306 KC614528 F3802 1004 Parque Estadual Intervales, SP 7 July 2009 −24.27638 −48.41556 GenBank KC608326 – Choeradoplana iheringi Graff, 1899 F3449 533 Floresta Nacional de São Francisco de Paula, RS 2 April 2012 −29.42367 −50.38711 GenBank KC608320 KC614540 Choeradoplana pucupucu Carbayo et al., 2017 F2840 540 Parque Nacional Serra da Bocaina, SP September 2008 −22.74277 −44.61639 GenBank KC608293 KC614517 Cratera crioula (E. M. Froehlich, 1955) F3708 1078 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608323 KC614543 F3715 1079 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608324 KC614544 Cratera pseudovaginuloides (Riester, 1938) F1244 670 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608251 KC614482 F1245 671 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608252 KC614483 Cratera tamoia (E. M. Froehlich, 1955) F1139 665 Parque Nacional da Serra dos Órgãos, RJ 15 January 2007 −22.45528 −42.99750 GenBank KC608246 KC614478 F1336 672 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608254 KC614484 Geobia subterranea (Schultze & Müller, 1857) F0358 650 Colégio Cristo Rei, São Leopoldo, RS 30 May 2004 −29.78828 −51.15084 GenBank KC608225 KC614464 F1355 673 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608255 KC614485 Geoplana apua Almeida & Carbayo sp. nov. F5236 2072 Nova Iguaçú, RJ 4–6 April 2012 Holotype MG653226* – Geoplana boraceia Almeida & Carbayo sp. nov. F4112 2070 Estação Biológica de Boraceia, SP 21 December 2009 −23.65389 −45.88917 Holotype – MG653220* F4358 1086 São Sebastião, SP 26 February 2010 −23.75203 −45.631027 Paratype KC608329 KC614547 F4389 2123 São Sebastião, SP 26 February 2010 −23.746 −45.632 Paratype MG653225* – F4409 1087 São Sebastião, SP 27 February 2010 −23.75203 −45.631027 External morphology similar to type material KC608330 KC614548 F6026 2095 São Sebastião, SP 11 January 2014 −23.74439 −45.63715 Paratype MG653227* MG653194* F6055 2125 São Sebastião, SP 11 January 2014 Paratype MG653237* MG653205* F6068 2124 São Sebastião, SP 12 January 2014 Paratype MG653244* – F6357 2073 São Sebastião, SP 27 September 2014 −23.74685 −45.63213 Paratype MG653238* MG653206* F6575 2077 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653239* MG653208* F6578 2078 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653240* MG653209* Geoplana cambara Almeida & Carbayo sp. nov. F1614 1009 Parque Nacional Saint Hilaire / Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank (named G. vaginuloides in GenBank) KC608262 KC614492 F1635 2063 Parque Nacional Saint Hilaire/Lange, PR 11 January 2008 Paratype MG653229* MG653198* Geoplana cananeia Almeida & Carbayo sp. nov. F4096 2067 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Holotype MG653223* MG653196* F4097 2068 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653233* MG653201* F4098 2069 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653224* – Geoplana caraguatatuba Almeida & Carbayo sp. nov. F6727 2080 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 May 2016 −23.594012 −45.429672 Paratype MG653247* – F6976 2126 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 January 2016 −23.593880 −45.429746 Paratype MG653251* – Geoplana chita Froehlich, 1956 F1710 715 Morretes, PR 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653218* F1730 716 Morretes, PR-25.469604 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653219* Geoplana ibiuna Almeida & Carbayo sp. nov. F3562 2066 Parque Estadual do Jurupará/Ibiúna, SP February 2009 Holotype MG653232* – F6907 2081 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653248* MG653214* F6908 2082 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653249* MG653215* F6909 2083 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653250* – Geoplana iporanga Almeida & Carbayo sp. nov. F3166 2065 Parque Estadual de Intervales, SP 13 December 2008 −24.269387 −48.405467 Holotype MG653231* MG653200* F3132 2064 Parque Estadual de Intervales/Ribeirão Grande, SP 13 December 2008 −24.269387 −48.405467 Paratype – MG653221* Geoplana mogi Almeida & Carbayo sp. nov. F1022 2061 Mogi das Cruzes, SP 27 May 2006 −23.451536 −46.227993 Paratype – MG653217* Geoplana paranapiacaba Almeida & Carbayo sp. nov. F6660 2079 Cidade Turística de Paranapiacaba, SP 5 June 2015 −23.77746 −46.30145 Holotype MG653246* MG653213* Geoplana piratininga Almeida & Carbayo sp. nov. F1163 666 Parque Jardim Previdência/SP 17 June 2007 −23.579662 −46.727503 Paratype KC608247 KC614479 F0509 2060 Parque Ecológico do Tietê/SP 27 May 2005 −23.490918 −46.516424 Paratype MG653222* MG653195* Geoplana pulchella Schultze & Müller, 1857 F3249 1068 Parque Estadual da Serra do Tabuleiro, SC 14 January 2009 −27.98139 −48.74806 External aspect similar to that of original description KC608310 KC614532 F6528 2075 RPPN Vale das Pedras, SC 16 January 2015 −27.61695 −049.34778 External aspect similar to that of original description MG653245* MG653212* F6544 2075 RPPN Vale das Pedras, SC 18 January 2015 −27.61696 −049.34813 External aspect similar to that of original description – MG653207* Geoplana quagga Marcus, 1951 F1003 1188 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 Genbank KC608236 – Geoplana sp. 1 F2934 2127 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653241* – F2943 2128 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653242* MG653210* F2944 2129 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653243* MG653211* Geoplana sp. 2 F5145 2130 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653234* MG653202* F5163 2131 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653235* MG653203* F5204 2132 Parque Nacional de Itatiaia, RJ 6 April 2012 Molecular and morphological support MG653236* MG653204* Geoplana sp. 3 F2638 2133 Parque Estadual de Intervales, SP 24 December 2008 −24.269387 −48.405467 Molecular support MG653230* MG653199* Geoplana sp. 4 F1707 2134 Morretes, PR 13 January 2008 −25.469604 −48.813739 Molecular support MG653252* MG653216* Geoplana vaginuloides (Darwin, 1844) F6387 2074 Parque Estadual da Pedra Branca, RJ 13 December 2014 −22.93707 −43.46098 Neotype MG653228* MG653197* Geoplana goetschi Riester, 1938, incertae sedis F2870 412 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608294 – Geoplana hina Marcus, 1951, (incertae sedis) F1613 1008 Parque Nacional Saint Hilaire/Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank KC608261 KC614491 Geoplana phocaica Marcus, 1951 (incertae sedis) F2827 457 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608289 KC614514 Geoplana phocaica Marcus, 1951 (incertae sedis) F2830 460 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608290 KC614515 Geoplaninae 1 F2033 1193 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.75000 −44.61666 GenBank KC608269 – Geoplaninae 2 F2807 1050 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 Named as Geoplana sp. 1 in GenBank KC608284 KC614510 F2821 1052 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 Named as Geoplana sp. 1 in GenBank KC608287 KC614513 Geoplaninae 3 F2783 1048 Parque Estadual Intervales, SP 29 July 2008 −24.27333 −48.41694 Named as Geoplana sp. 2 in GenBank KC608281 KC614508 Geoplaninae 4 F2796 1194 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 3 in GenBank KC608282 – Geoplaninae 5 F2031 1014 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.73333 −44.60000 Named as Geoplana sp. 4 in GenBank KC608268 KC614497 F2809 1051 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 4 in GenBank KC608285 KC614511 Geoplaninae 6 F2189 348 Parque Estadual do Desengano, RJ 18 March 2008 −21.87667 −41.91972 Named as Geoplana sp. 5 in GenBank KC608278 KC614505 Geoplaninae 7 F3191 1066 Parque Estadual da Serra do Tabuleiro, SC 9 January 2009 −27.95989 −48.75761 Named as Geoplana sp. 6 in GenBank KC608308 KC614530 F3247 1067 Parque Estadual da Serra do Tabuleiro, SC 13 January 2009 −27.98139 −48.74806 Named as Geoplana sp. 6 in GenBank KC608309 KC614531 Gusana sp. 1 F4421 1088 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608331 KC614549 F4428 1089 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608332 – Imbira guaiana (Leal-Zanchet & Carbayo, 2001) F0432 653 Floresta Nacional de São Francisco de Paula, RS 1 August 2004 −29.41666 −50.38333 GenBank KC608229 KC614467 F3361 1071 Floresta Nacional de São Francisco de Paula, RS 20 January 2009 −29.42367 −50.38711 GenBank KC608314 – Imbira marcusi Carbayo et al., 2013 F1041 1190 São Sebastião, SP 8 September 2006 −23.75203 −45.631027 GenBank KC608241 – F2833 404 Parque Nacional Serra da Bocaina, SP September 2008 −22.75222 −44.62194 GenBank KC608291 KC614516 Issoca jandaia Froehlich, 1955 F2043 1015 Parque Nacional Serra da Bocaina, SP 9 February 2007 −22.78222 −44.60583 GenBank KC608270 KC614498 Issoca rezendei (Schirch, 1929) F1182 667 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608248 KC614480 F1679 1010 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608263 KC614493 Issoca sp. 1 F2158 1020 Parque Estadual do Desengano, RJ 16 March 2008 −21.87694 −41.92583 GenBank KC608275 KC614502 F2507 1045 Reserva Biológica Augusto Ruschi, ES 24 May 2008 −19.88740 −40.54319 GenBank KC608279 KC614506 F4085 1085 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608328 KC614546 Luteostriata abundans (Graff, 1899) no voucher São Leopoldo, RS 1998 −29.78828 −51.15084 GenBank KC631621 KC614556 F0205 646 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608223 KC614462 F0238 648 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608224 KC614463 Luteostriata ceciliae (Froehlich & Leal-Zanchet, 2003) F3451 1077 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608321 KC614541 Luteostriata ernesti (Leal-Zanchet & Froehlich, 2006) no voucher São Paulo, SP 2005 −23.49080 −46.52090 GenBank – KC614555 F3358 1070 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608313 KC614534 Luteostriata graffi (Leal-Zanchet & Froehlich, 2006) F0384 652 Taquara, RS 28 January 2010 −29.660524 −50.746782 GenBank KC608227 KC614466 F0440 654 Morro das Aranhas, Florianópolis, SC 9 September 2004 −27.468485 −48.377724 GenBank KC608230 KC614468 F3381 1072 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42963 −50.39780 GenBank KC608315 KC614535 Luteostriata muelleri (Diesing, 1861) No voucher GenBank – KC614554 F3268 1199 Parque Estadual da Serra do Tabuleiro, SC 15 January 2009 −27.84278 −48.92583 GenBank KC608311 – Luteostriata sp. F1192 668 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608249 KC614481 Luteostriata sp. 1 F1036 659 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608239 KC614473 F1078 663 Estação Biológica de Boraceia, Salesópolis, SP, Brazil 2 February 2006 −23.65388 −45.88910 GenBank KC608243 KC614476 Luteostriata sp. 2 F0441 655 Santinhos, Florianópolis, SC 9 September 2004 −27.46370 −48.38100 GenBank KC608231 KC614469 Matuxia matuta (E. M. Froehlich, 1955) F2184 1021 Parque Estadual do Desengano, RJ 17 March 2008 −27.95989 −48.75761 GenBank KC608276 KC614503 F2187 1022 Parque Estadual do Desengano, RJ 17 March 2008 −21.87138 −41.91472 GenBank KC608277 KC614504 Matuxia tuxaua E. M. Froehlich, 1955 F1275 1192 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608253 – F3059 1058 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608302 KC614525 Notogynaphallia albonigra (Riester, 1938) F4081 1083 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608327 KC614545 Notogynaphallia parca (E. M. Froehlich, 1955) F2972 1196 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.42914 −46.63250 GenBank KC608298 – Notogynaphallia sexstriata (Graff, 1899) F0792 656 Cidade Universitária, USP, São Paulo, SP 19 December 2005 −23.56671 −46.73017 GenBank KC608232 KC614470 F1413 680 Parque Nacional da Serra de Itajaí, SC 4 January 2008 −27.04836 −49.09200 GenBank KC608257 KC614487 Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015 F1399 1007 Parque Nacional da Serra do Itajaí, Blumenau, SC 4 January 2008 −27.05306 −49.08528 GenBank KC608256 KC614486 F1504 698 Parque Nacional da Serra do Itajaí, Blumenau, SC 6 January 2008 −27.05306 −49.08528 GenBank KC608259 KC614489 F1566 707 Parque Nacional da Serra do Itajaí, Blumenau, SC 7 January 2008 −27.05306 −49.08528 GenBank KC608260 KC614490 Obama burmeisteri (Schultze & Müller, 1857) F0989 658 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608234 – F2886 733 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608297 KC614520 Obama carinata (Riester, 1938) F3084 1062 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608304 KC614527 F3093 1064 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608307 KC614529 Obama josefi (Carbayo & Leal-Zanchet, 2001) F3389 1074 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608317 KC614537 F3403 1075 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608318 KC614538 Obama ladislavii (Graff, 1899) F1418 681 Parque Nacional da Serra de Itajaí, SC 5 January 2008 −27.04836 −49.09200 GenBank KC608258 KC614488 Paraba franciscana (Leal-Zanchet & Carbayo, 2001) F3357 1069 Floresta Nacional de São Francisco de Paula, RS 2 November 2006 −29.42894 −50.39200 GenBank KC608312 KC614533 F5485 935 Floresta Nacional de São Francisco de Paula, RS 12 August 1999 −29.36666 −50.38333 GenBank KC608335 KC614552 Paraba multicolor (Graff, 1899) F1736 594 Parque Nacional Saint Hilaire/Lange, PR 14 January 2008 −25.46972 −48.81222 GenBank KC608265 KC614495 F2997 1055 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608299 KC614521 Paraba rubidolineata (Baptista & Leal-Zanchet, 2005) F5482 936 Floresta Nacional de São Francisco de Paula, RS 26 August 1999 −29.36666 −50.38333 GenBank KC608333 KC614550 Pasipha chimbeva (E. M. Froehlich, 1955) F2813 403 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 GenBank KC608286 KC614512 F3010 476 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608300 KC614522 Pasipha pasipha (Marcus, 1951) F1841 1012 Parque Estadual Alberto Löfgren, São Paulo, SP 26 January 2008 −23.45925 −46.63730 GenBank KC608266 KC614496 F2871 1053 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608295 KC614518 Pasipha pinima (E. M. Froehlich, 1955) F1714 1011 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608264 KC614494 F2585 717 Parque Nacional da Serra dos Órgãos, RJ 10 July 2008 −22.45528 −42.99750 GenBank KC608280 KC614507 Pasipha rosea (E. M. Froehlich, 1955) F1061 660 Parque Estoril, São Bernardo do Campo, SP 22 October 2006 −23.77368 −46.51469 GenBank KC608242 KC614475 Pasipha sp. 2 F1138 664 Parque Nacional da Serra dos Órgãos, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608245 KC614477 Pasipha tapetilla (Marcus, 1951) F2878 732 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608296 KC614519 F5631B 938 São Paulo, SP May 2007 −29.36666 −50.38333 GenBank KC608336 KC614553 Polycladus sp. 1 F0397 1186 Valdívia (Chile) 2004 −39.81000 −73.23000 GenBank KC608228 – Supramontana irritata Carbayo & Leal-Zanchet, 2003 F3452 772 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42064 −50.39394 GenBank KC608322 KC614542 F5483 937 Floresta Nacional de São Francisco de Paula, RS 9 February 2006 −29.36666 −50.38333 GenBank KC608334 KC614551 Xerapoa hystrix Froehlich, 1955 F3057 1057 Parque Estadual Serra da Cantareira, SP 12 December 2008 −24.27638 −48.41556 GenBank – KC614524 Xerapoa pseudorhynchodemus (Riester, 1938) F0992 1187 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608235 – F1013 1189 Cidade Universitária, USP, São Paulo, SP 17 March 2006 −23.56671 −46.73017 GenBank KC608237 – Xerapoa trina (Marcus, 1951) F1134 1191 Cascata Imbuí, Teresópolis, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608244 – F1239 669 Parque Nacional da Serra dos Órgãos, RJ 15 December 2007 −22.45528 −42.99750 GenBank KC608250 – *All sequences obtained for this study. All specimens were collected in Brazil, unless specified otherwise. Abbreviations for the Brazilian States are as follows: PR, Paraná; RJ, Rio de Janeiro; RS, Rio Grande do Sul; SC, Santa Catarina; SP, São Paulo. View Large Table 1. Individuals used in the study, museum code used in the figures and text, collecting locality data, identification base and GenBank accession numbers for all sequences Species Field number Specimen’s accession number (MZUSP PL) Collecting locality Collecting date Latitude Longitude Identification COI EF-1-alpha Rhynchodeminae Arthurdendyus triangulatus (Dendy, 1895) GenBank DQ666027 KC614559 Artioposthia testacea Hutton, 1880 GenBank AF178305 KC614560 Australoplana sp. GenBank DQ666028 KC614558 Dolichoplana striata Moseley, 1877 F0379 1005 Igrejinha, RS 10 July 2004 −29.57200 −50.77800 GenBank KC608226 KC614465 Endeavouria septemlineata (Hyman, 1939) F0988 657 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608233 KC614471 F0192 1184 Maquiné, RS 10 February 2004 −29.67774 −50.20640 GenBank KC608222 KC614461 Geoplaninae Cephaloflexa araucariana Carbayo & Leal-Zanchet, 2003 F3387 1073 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608316 KC614536 F3426 1076 Floresta Nacional de São Francisco de Paula, RS 23 January 2009 −29.42739 −50.39072 GenBank KC608319 KC614539 Cephaloflexa bergi (Graff, 1899) F1034 303 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608238 KC614472 F1038 305 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608240 KC614474 Cephaloflexa sp. 1 F2103 1211 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608272 – F2108 1019 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608274 KC614501 Choeradoplana banga Carbayo & E. M. Froehlich, 2012 F2023 1000 Parque Estadual Serra da Cantareira, SP 30 January 2008 −23.42916 −46.63250 GenBank KC608267 – F3011 1002 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608301 KC614523 Choeradoplana bocaina Carbayo & E. M. Froehlich, 2012 F2104 999 Parque Nacional Serra da Bocaina, SP 10 February 2008 −22.75222 −44.62194 GenBank KC608273 KC614500 F2803 998 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 GenBank KC608283 KC614509 F2822 997 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 GenBank KC608288 – Choeradoplana gladismariae Carbayo & E. M. Froehlich, 2012 F3092 1003 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608306 KC614528 F3802 1004 Parque Estadual Intervales, SP 7 July 2009 −24.27638 −48.41556 GenBank KC608326 – Choeradoplana iheringi Graff, 1899 F3449 533 Floresta Nacional de São Francisco de Paula, RS 2 April 2012 −29.42367 −50.38711 GenBank KC608320 KC614540 Choeradoplana pucupucu Carbayo et al., 2017 F2840 540 Parque Nacional Serra da Bocaina, SP September 2008 −22.74277 −44.61639 GenBank KC608293 KC614517 Cratera crioula (E. M. Froehlich, 1955) F3708 1078 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608323 KC614543 F3715 1079 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608324 KC614544 Cratera pseudovaginuloides (Riester, 1938) F1244 670 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608251 KC614482 F1245 671 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608252 KC614483 Cratera tamoia (E. M. Froehlich, 1955) F1139 665 Parque Nacional da Serra dos Órgãos, RJ 15 January 2007 −22.45528 −42.99750 GenBank KC608246 KC614478 F1336 672 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608254 KC614484 Geobia subterranea (Schultze & Müller, 1857) F0358 650 Colégio Cristo Rei, São Leopoldo, RS 30 May 2004 −29.78828 −51.15084 GenBank KC608225 KC614464 F1355 673 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608255 KC614485 Geoplana apua Almeida & Carbayo sp. nov. F5236 2072 Nova Iguaçú, RJ 4–6 April 2012 Holotype MG653226* – Geoplana boraceia Almeida & Carbayo sp. nov. F4112 2070 Estação Biológica de Boraceia, SP 21 December 2009 −23.65389 −45.88917 Holotype – MG653220* F4358 1086 São Sebastião, SP 26 February 2010 −23.75203 −45.631027 Paratype KC608329 KC614547 F4389 2123 São Sebastião, SP 26 February 2010 −23.746 −45.632 Paratype MG653225* – F4409 1087 São Sebastião, SP 27 February 2010 −23.75203 −45.631027 External morphology similar to type material KC608330 KC614548 F6026 2095 São Sebastião, SP 11 January 2014 −23.74439 −45.63715 Paratype MG653227* MG653194* F6055 2125 São Sebastião, SP 11 January 2014 Paratype MG653237* MG653205* F6068 2124 São Sebastião, SP 12 January 2014 Paratype MG653244* – F6357 2073 São Sebastião, SP 27 September 2014 −23.74685 −45.63213 Paratype MG653238* MG653206* F6575 2077 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653239* MG653208* F6578 2078 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653240* MG653209* Geoplana cambara Almeida & Carbayo sp. nov. F1614 1009 Parque Nacional Saint Hilaire / Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank (named G. vaginuloides in GenBank) KC608262 KC614492 F1635 2063 Parque Nacional Saint Hilaire/Lange, PR 11 January 2008 Paratype MG653229* MG653198* Geoplana cananeia Almeida & Carbayo sp. nov. F4096 2067 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Holotype MG653223* MG653196* F4097 2068 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653233* MG653201* F4098 2069 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653224* – Geoplana caraguatatuba Almeida & Carbayo sp. nov. F6727 2080 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 May 2016 −23.594012 −45.429672 Paratype MG653247* – F6976 2126 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 January 2016 −23.593880 −45.429746 Paratype MG653251* – Geoplana chita Froehlich, 1956 F1710 715 Morretes, PR 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653218* F1730 716 Morretes, PR-25.469604 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653219* Geoplana ibiuna Almeida & Carbayo sp. nov. F3562 2066 Parque Estadual do Jurupará/Ibiúna, SP February 2009 Holotype MG653232* – F6907 2081 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653248* MG653214* F6908 2082 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653249* MG653215* F6909 2083 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653250* – Geoplana iporanga Almeida & Carbayo sp. nov. F3166 2065 Parque Estadual de Intervales, SP 13 December 2008 −24.269387 −48.405467 Holotype MG653231* MG653200* F3132 2064 Parque Estadual de Intervales/Ribeirão Grande, SP 13 December 2008 −24.269387 −48.405467 Paratype – MG653221* Geoplana mogi Almeida & Carbayo sp. nov. F1022 2061 Mogi das Cruzes, SP 27 May 2006 −23.451536 −46.227993 Paratype – MG653217* Geoplana paranapiacaba Almeida & Carbayo sp. nov. F6660 2079 Cidade Turística de Paranapiacaba, SP 5 June 2015 −23.77746 −46.30145 Holotype MG653246* MG653213* Geoplana piratininga Almeida & Carbayo sp. nov. F1163 666 Parque Jardim Previdência/SP 17 June 2007 −23.579662 −46.727503 Paratype KC608247 KC614479 F0509 2060 Parque Ecológico do Tietê/SP 27 May 2005 −23.490918 −46.516424 Paratype MG653222* MG653195* Geoplana pulchella Schultze & Müller, 1857 F3249 1068 Parque Estadual da Serra do Tabuleiro, SC 14 January 2009 −27.98139 −48.74806 External aspect similar to that of original description KC608310 KC614532 F6528 2075 RPPN Vale das Pedras, SC 16 January 2015 −27.61695 −049.34778 External aspect similar to that of original description MG653245* MG653212* F6544 2075 RPPN Vale das Pedras, SC 18 January 2015 −27.61696 −049.34813 External aspect similar to that of original description – MG653207* Geoplana quagga Marcus, 1951 F1003 1188 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 Genbank KC608236 – Geoplana sp. 1 F2934 2127 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653241* – F2943 2128 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653242* MG653210* F2944 2129 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653243* MG653211* Geoplana sp. 2 F5145 2130 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653234* MG653202* F5163 2131 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653235* MG653203* F5204 2132 Parque Nacional de Itatiaia, RJ 6 April 2012 Molecular and morphological support MG653236* MG653204* Geoplana sp. 3 F2638 2133 Parque Estadual de Intervales, SP 24 December 2008 −24.269387 −48.405467 Molecular support MG653230* MG653199* Geoplana sp. 4 F1707 2134 Morretes, PR 13 January 2008 −25.469604 −48.813739 Molecular support MG653252* MG653216* Geoplana vaginuloides (Darwin, 1844) F6387 2074 Parque Estadual da Pedra Branca, RJ 13 December 2014 −22.93707 −43.46098 Neotype MG653228* MG653197* Geoplana goetschi Riester, 1938, incertae sedis F2870 412 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608294 – Geoplana hina Marcus, 1951, (incertae sedis) F1613 1008 Parque Nacional Saint Hilaire/Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank KC608261 KC614491 Geoplana phocaica Marcus, 1951 (incertae sedis) F2827 457 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608289 KC614514 Geoplana phocaica Marcus, 1951 (incertae sedis) F2830 460 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608290 KC614515 Geoplaninae 1 F2033 1193 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.75000 −44.61666 GenBank KC608269 – Geoplaninae 2 F2807 1050 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 Named as Geoplana sp. 1 in GenBank KC608284 KC614510 F2821 1052 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 Named as Geoplana sp. 1 in GenBank KC608287 KC614513 Geoplaninae 3 F2783 1048 Parque Estadual Intervales, SP 29 July 2008 −24.27333 −48.41694 Named as Geoplana sp. 2 in GenBank KC608281 KC614508 Geoplaninae 4 F2796 1194 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 3 in GenBank KC608282 – Geoplaninae 5 F2031 1014 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.73333 −44.60000 Named as Geoplana sp. 4 in GenBank KC608268 KC614497 F2809 1051 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 4 in GenBank KC608285 KC614511 Geoplaninae 6 F2189 348 Parque Estadual do Desengano, RJ 18 March 2008 −21.87667 −41.91972 Named as Geoplana sp. 5 in GenBank KC608278 KC614505 Geoplaninae 7 F3191 1066 Parque Estadual da Serra do Tabuleiro, SC 9 January 2009 −27.95989 −48.75761 Named as Geoplana sp. 6 in GenBank KC608308 KC614530 F3247 1067 Parque Estadual da Serra do Tabuleiro, SC 13 January 2009 −27.98139 −48.74806 Named as Geoplana sp. 6 in GenBank KC608309 KC614531 Gusana sp. 1 F4421 1088 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608331 KC614549 F4428 1089 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608332 – Imbira guaiana (Leal-Zanchet & Carbayo, 2001) F0432 653 Floresta Nacional de São Francisco de Paula, RS 1 August 2004 −29.41666 −50.38333 GenBank KC608229 KC614467 F3361 1071 Floresta Nacional de São Francisco de Paula, RS 20 January 2009 −29.42367 −50.38711 GenBank KC608314 – Imbira marcusi Carbayo et al., 2013 F1041 1190 São Sebastião, SP 8 September 2006 −23.75203 −45.631027 GenBank KC608241 – F2833 404 Parque Nacional Serra da Bocaina, SP September 2008 −22.75222 −44.62194 GenBank KC608291 KC614516 Issoca jandaia Froehlich, 1955 F2043 1015 Parque Nacional Serra da Bocaina, SP 9 February 2007 −22.78222 −44.60583 GenBank KC608270 KC614498 Issoca rezendei (Schirch, 1929) F1182 667 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608248 KC614480 F1679 1010 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608263 KC614493 Issoca sp. 1 F2158 1020 Parque Estadual do Desengano, RJ 16 March 2008 −21.87694 −41.92583 GenBank KC608275 KC614502 F2507 1045 Reserva Biológica Augusto Ruschi, ES 24 May 2008 −19.88740 −40.54319 GenBank KC608279 KC614506 F4085 1085 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608328 KC614546 Luteostriata abundans (Graff, 1899) no voucher São Leopoldo, RS 1998 −29.78828 −51.15084 GenBank KC631621 KC614556 F0205 646 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608223 KC614462 F0238 648 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608224 KC614463 Luteostriata ceciliae (Froehlich & Leal-Zanchet, 2003) F3451 1077 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608321 KC614541 Luteostriata ernesti (Leal-Zanchet & Froehlich, 2006) no voucher São Paulo, SP 2005 −23.49080 −46.52090 GenBank – KC614555 F3358 1070 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608313 KC614534 Luteostriata graffi (Leal-Zanchet & Froehlich, 2006) F0384 652 Taquara, RS 28 January 2010 −29.660524 −50.746782 GenBank KC608227 KC614466 F0440 654 Morro das Aranhas, Florianópolis, SC 9 September 2004 −27.468485 −48.377724 GenBank KC608230 KC614468 F3381 1072 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42963 −50.39780 GenBank KC608315 KC614535 Luteostriata muelleri (Diesing, 1861) No voucher GenBank – KC614554 F3268 1199 Parque Estadual da Serra do Tabuleiro, SC 15 January 2009 −27.84278 −48.92583 GenBank KC608311 – Luteostriata sp. F1192 668 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608249 KC614481 Luteostriata sp. 1 F1036 659 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608239 KC614473 F1078 663 Estação Biológica de Boraceia, Salesópolis, SP, Brazil 2 February 2006 −23.65388 −45.88910 GenBank KC608243 KC614476 Luteostriata sp. 2 F0441 655 Santinhos, Florianópolis, SC 9 September 2004 −27.46370 −48.38100 GenBank KC608231 KC614469 Matuxia matuta (E. M. Froehlich, 1955) F2184 1021 Parque Estadual do Desengano, RJ 17 March 2008 −27.95989 −48.75761 GenBank KC608276 KC614503 F2187 1022 Parque Estadual do Desengano, RJ 17 March 2008 −21.87138 −41.91472 GenBank KC608277 KC614504 Matuxia tuxaua E. M. Froehlich, 1955 F1275 1192 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608253 – F3059 1058 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608302 KC614525 Notogynaphallia albonigra (Riester, 1938) F4081 1083 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608327 KC614545 Notogynaphallia parca (E. M. Froehlich, 1955) F2972 1196 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.42914 −46.63250 GenBank KC608298 – Notogynaphallia sexstriata (Graff, 1899) F0792 656 Cidade Universitária, USP, São Paulo, SP 19 December 2005 −23.56671 −46.73017 GenBank KC608232 KC614470 F1413 680 Parque Nacional da Serra de Itajaí, SC 4 January 2008 −27.04836 −49.09200 GenBank KC608257 KC614487 Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015 F1399 1007 Parque Nacional da Serra do Itajaí, Blumenau, SC 4 January 2008 −27.05306 −49.08528 GenBank KC608256 KC614486 F1504 698 Parque Nacional da Serra do Itajaí, Blumenau, SC 6 January 2008 −27.05306 −49.08528 GenBank KC608259 KC614489 F1566 707 Parque Nacional da Serra do Itajaí, Blumenau, SC 7 January 2008 −27.05306 −49.08528 GenBank KC608260 KC614490 Obama burmeisteri (Schultze & Müller, 1857) F0989 658 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608234 – F2886 733 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608297 KC614520 Obama carinata (Riester, 1938) F3084 1062 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608304 KC614527 F3093 1064 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608307 KC614529 Obama josefi (Carbayo & Leal-Zanchet, 2001) F3389 1074 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608317 KC614537 F3403 1075 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608318 KC614538 Obama ladislavii (Graff, 1899) F1418 681 Parque Nacional da Serra de Itajaí, SC 5 January 2008 −27.04836 −49.09200 GenBank KC608258 KC614488 Paraba franciscana (Leal-Zanchet & Carbayo, 2001) F3357 1069 Floresta Nacional de São Francisco de Paula, RS 2 November 2006 −29.42894 −50.39200 GenBank KC608312 KC614533 F5485 935 Floresta Nacional de São Francisco de Paula, RS 12 August 1999 −29.36666 −50.38333 GenBank KC608335 KC614552 Paraba multicolor (Graff, 1899) F1736 594 Parque Nacional Saint Hilaire/Lange, PR 14 January 2008 −25.46972 −48.81222 GenBank KC608265 KC614495 F2997 1055 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608299 KC614521 Paraba rubidolineata (Baptista & Leal-Zanchet, 2005) F5482 936 Floresta Nacional de São Francisco de Paula, RS 26 August 1999 −29.36666 −50.38333 GenBank KC608333 KC614550 Pasipha chimbeva (E. M. Froehlich, 1955) F2813 403 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 GenBank KC608286 KC614512 F3010 476 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608300 KC614522 Pasipha pasipha (Marcus, 1951) F1841 1012 Parque Estadual Alberto Löfgren, São Paulo, SP 26 January 2008 −23.45925 −46.63730 GenBank KC608266 KC614496 F2871 1053 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608295 KC614518 Pasipha pinima (E. M. Froehlich, 1955) F1714 1011 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608264 KC614494 F2585 717 Parque Nacional da Serra dos Órgãos, RJ 10 July 2008 −22.45528 −42.99750 GenBank KC608280 KC614507 Pasipha rosea (E. M. Froehlich, 1955) F1061 660 Parque Estoril, São Bernardo do Campo, SP 22 October 2006 −23.77368 −46.51469 GenBank KC608242 KC614475 Pasipha sp. 2 F1138 664 Parque Nacional da Serra dos Órgãos, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608245 KC614477 Pasipha tapetilla (Marcus, 1951) F2878 732 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608296 KC614519 F5631B 938 São Paulo, SP May 2007 −29.36666 −50.38333 GenBank KC608336 KC614553 Polycladus sp. 1 F0397 1186 Valdívia (Chile) 2004 −39.81000 −73.23000 GenBank KC608228 – Supramontana irritata Carbayo & Leal-Zanchet, 2003 F3452 772 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42064 −50.39394 GenBank KC608322 KC614542 F5483 937 Floresta Nacional de São Francisco de Paula, RS 9 February 2006 −29.36666 −50.38333 GenBank KC608334 KC614551 Xerapoa hystrix Froehlich, 1955 F3057 1057 Parque Estadual Serra da Cantareira, SP 12 December 2008 −24.27638 −48.41556 GenBank – KC614524 Xerapoa pseudorhynchodemus (Riester, 1938) F0992 1187 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608235 – F1013 1189 Cidade Universitária, USP, São Paulo, SP 17 March 2006 −23.56671 −46.73017 GenBank KC608237 – Xerapoa trina (Marcus, 1951) F1134 1191 Cascata Imbuí, Teresópolis, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608244 – F1239 669 Parque Nacional da Serra dos Órgãos, RJ 15 December 2007 −22.45528 −42.99750 GenBank KC608250 – Species Field number Specimen’s accession number (MZUSP PL) Collecting locality Collecting date Latitude Longitude Identification COI EF-1-alpha Rhynchodeminae Arthurdendyus triangulatus (Dendy, 1895) GenBank DQ666027 KC614559 Artioposthia testacea Hutton, 1880 GenBank AF178305 KC614560 Australoplana sp. GenBank DQ666028 KC614558 Dolichoplana striata Moseley, 1877 F0379 1005 Igrejinha, RS 10 July 2004 −29.57200 −50.77800 GenBank KC608226 KC614465 Endeavouria septemlineata (Hyman, 1939) F0988 657 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608233 KC614471 F0192 1184 Maquiné, RS 10 February 2004 −29.67774 −50.20640 GenBank KC608222 KC614461 Geoplaninae Cephaloflexa araucariana Carbayo & Leal-Zanchet, 2003 F3387 1073 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608316 KC614536 F3426 1076 Floresta Nacional de São Francisco de Paula, RS 23 January 2009 −29.42739 −50.39072 GenBank KC608319 KC614539 Cephaloflexa bergi (Graff, 1899) F1034 303 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608238 KC614472 F1038 305 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608240 KC614474 Cephaloflexa sp. 1 F2103 1211 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608272 – F2108 1019 Parque Nacional Serra da Bocaina, SP 10 February 2008 −23.65388 −45.88910 GenBank KC608274 KC614501 Choeradoplana banga Carbayo & E. M. Froehlich, 2012 F2023 1000 Parque Estadual Serra da Cantareira, SP 30 January 2008 −23.42916 −46.63250 GenBank KC608267 – F3011 1002 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608301 KC614523 Choeradoplana bocaina Carbayo & E. M. Froehlich, 2012 F2104 999 Parque Nacional Serra da Bocaina, SP 10 February 2008 −22.75222 −44.62194 GenBank KC608273 KC614500 F2803 998 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 GenBank KC608283 KC614509 F2822 997 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 GenBank KC608288 – Choeradoplana gladismariae Carbayo & E. M. Froehlich, 2012 F3092 1003 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608306 KC614528 F3802 1004 Parque Estadual Intervales, SP 7 July 2009 −24.27638 −48.41556 GenBank KC608326 – Choeradoplana iheringi Graff, 1899 F3449 533 Floresta Nacional de São Francisco de Paula, RS 2 April 2012 −29.42367 −50.38711 GenBank KC608320 KC614540 Choeradoplana pucupucu Carbayo et al., 2017 F2840 540 Parque Nacional Serra da Bocaina, SP September 2008 −22.74277 −44.61639 GenBank KC608293 KC614517 Cratera crioula (E. M. Froehlich, 1955) F3708 1078 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608323 KC614543 F3715 1079 Parque Estadual Serra da Cantareira, SP 19 April 2009 −23.43084 −46.63413 GenBank KC608324 KC614544 Cratera pseudovaginuloides (Riester, 1938) F1244 670 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608251 KC614482 F1245 671 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608252 KC614483 Cratera tamoia (E. M. Froehlich, 1955) F1139 665 Parque Nacional da Serra dos Órgãos, RJ 15 January 2007 −22.45528 −42.99750 GenBank KC608246 KC614478 F1336 672 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608254 KC614484 Geobia subterranea (Schultze & Müller, 1857) F0358 650 Colégio Cristo Rei, São Leopoldo, RS 30 May 2004 −29.78828 −51.15084 GenBank KC608225 KC614464 F1355 673 Parque Nacional da Serra dos Órgãos, RJ 18 December 2007 −22.49870 −42.99620 GenBank KC608255 KC614485 Geoplana apua Almeida & Carbayo sp. nov. F5236 2072 Nova Iguaçú, RJ 4–6 April 2012 Holotype MG653226* – Geoplana boraceia Almeida & Carbayo sp. nov. F4112 2070 Estação Biológica de Boraceia, SP 21 December 2009 −23.65389 −45.88917 Holotype – MG653220* F4358 1086 São Sebastião, SP 26 February 2010 −23.75203 −45.631027 Paratype KC608329 KC614547 F4389 2123 São Sebastião, SP 26 February 2010 −23.746 −45.632 Paratype MG653225* – F4409 1087 São Sebastião, SP 27 February 2010 −23.75203 −45.631027 External morphology similar to type material KC608330 KC614548 F6026 2095 São Sebastião, SP 11 January 2014 −23.74439 −45.63715 Paratype MG653227* MG653194* F6055 2125 São Sebastião, SP 11 January 2014 Paratype MG653237* MG653205* F6068 2124 São Sebastião, SP 12 January 2014 Paratype MG653244* – F6357 2073 São Sebastião, SP 27 September 2014 −23.74685 −45.63213 Paratype MG653238* MG653206* F6575 2077 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653239* MG653208* F6578 2078 São Sebastião, SP 13 February 2015 −23.74685 −45.63213 Paratype MG653240* MG653209* Geoplana cambara Almeida & Carbayo sp. nov. F1614 1009 Parque Nacional Saint Hilaire / Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank (named G. vaginuloides in GenBank) KC608262 KC614492 F1635 2063 Parque Nacional Saint Hilaire/Lange, PR 11 January 2008 Paratype MG653229* MG653198* Geoplana cananeia Almeida & Carbayo sp. nov. F4096 2067 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Holotype MG653223* MG653196* F4097 2068 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653233* MG653201* F4098 2069 Parque Estadual da Ilha do Cardoso/Cananeia, SP 18 October 2009 −25.091235 −47.928957 Paratype MG653224* – Geoplana caraguatatuba Almeida & Carbayo sp. nov. F6727 2080 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 May 2016 −23.594012 −45.429672 Paratype MG653247* – F6976 2126 Parque Estadual Serra do Mar/Caraguatatuba, SP 14 January 2016 −23.593880 −45.429746 Paratype MG653251* – Geoplana chita Froehlich, 1956 F1710 715 Morretes, PR 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653218* F1730 716 Morretes, PR-25.469604 13 January 2008 −25.469604 −48.813739 External and internal morphology similar to that of type material – MG653219* Geoplana ibiuna Almeida & Carbayo sp. nov. F3562 2066 Parque Estadual do Jurupará/Ibiúna, SP February 2009 Holotype MG653232* – F6907 2081 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653248* MG653214* F6908 2082 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653249* MG653215* F6909 2083 Santana de Parnaíba, SP −23.449229 −47.012175 Paratype MG653250* – Geoplana iporanga Almeida & Carbayo sp. nov. F3166 2065 Parque Estadual de Intervales, SP 13 December 2008 −24.269387 −48.405467 Holotype MG653231* MG653200* F3132 2064 Parque Estadual de Intervales/Ribeirão Grande, SP 13 December 2008 −24.269387 −48.405467 Paratype – MG653221* Geoplana mogi Almeida & Carbayo sp. nov. F1022 2061 Mogi das Cruzes, SP 27 May 2006 −23.451536 −46.227993 Paratype – MG653217* Geoplana paranapiacaba Almeida & Carbayo sp. nov. F6660 2079 Cidade Turística de Paranapiacaba, SP 5 June 2015 −23.77746 −46.30145 Holotype MG653246* MG653213* Geoplana piratininga Almeida & Carbayo sp. nov. F1163 666 Parque Jardim Previdência/SP 17 June 2007 −23.579662 −46.727503 Paratype KC608247 KC614479 F0509 2060 Parque Ecológico do Tietê/SP 27 May 2005 −23.490918 −46.516424 Paratype MG653222* MG653195* Geoplana pulchella Schultze & Müller, 1857 F3249 1068 Parque Estadual da Serra do Tabuleiro, SC 14 January 2009 −27.98139 −48.74806 External aspect similar to that of original description KC608310 KC614532 F6528 2075 RPPN Vale das Pedras, SC 16 January 2015 −27.61695 −049.34778 External aspect similar to that of original description MG653245* MG653212* F6544 2075 RPPN Vale das Pedras, SC 18 January 2015 −27.61696 −049.34813 External aspect similar to that of original description – MG653207* Geoplana quagga Marcus, 1951 F1003 1188 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 Genbank KC608236 – Geoplana sp. 1 F2934 2127 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653241* – F2943 2128 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653242* MG653210* F2944 2129 Parque Estadual da Cantareira, SP 26 October 2008 Molecular and morphological support MG653243* MG653211* Geoplana sp. 2 F5145 2130 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653234* MG653202* F5163 2131 Parque Nacional de Itatiaia, RJ 4 April 2012 Molecular and morphological support MG653235* MG653203* F5204 2132 Parque Nacional de Itatiaia, RJ 6 April 2012 Molecular and morphological support MG653236* MG653204* Geoplana sp. 3 F2638 2133 Parque Estadual de Intervales, SP 24 December 2008 −24.269387 −48.405467 Molecular support MG653230* MG653199* Geoplana sp. 4 F1707 2134 Morretes, PR 13 January 2008 −25.469604 −48.813739 Molecular support MG653252* MG653216* Geoplana vaginuloides (Darwin, 1844) F6387 2074 Parque Estadual da Pedra Branca, RJ 13 December 2014 −22.93707 −43.46098 Neotype MG653228* MG653197* Geoplana goetschi Riester, 1938, incertae sedis F2870 412 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608294 – Geoplana hina Marcus, 1951, (incertae sedis) F1613 1008 Parque Nacional Saint Hilaire/Lange, PR 10 January 2008 −25.76437 −48.62266 GenBank KC608261 KC614491 Geoplana phocaica Marcus, 1951 (incertae sedis) F2827 457 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608289 KC614514 Geoplana phocaica Marcus, 1951 (incertae sedis) F2830 460 Parque Nacional Serra da Bocaina, SP September 2008 −22.73389 −44.61639 GenBank KC608290 KC614515 Geoplaninae 1 F2033 1193 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.75000 −44.61666 GenBank KC608269 – Geoplaninae 2 F2807 1050 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.75222 −44.62194 Named as Geoplana sp. 1 in GenBank KC608284 KC614510 F2821 1052 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.75000 −44.61666 Named as Geoplana sp. 1 in GenBank KC608287 KC614513 Geoplaninae 3 F2783 1048 Parque Estadual Intervales, SP 29 July 2008 −24.27333 −48.41694 Named as Geoplana sp. 2 in GenBank KC608281 KC614508 Geoplaninae 4 F2796 1194 Parque Nacional Serra da Bocaina, SP 7 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 3 in GenBank KC608282 – Geoplaninae 5 F2031 1014 Parque Nacional Serra da Bocaina, SP 7 February 2008 −22.73333 −44.60000 Named as Geoplana sp. 4 in GenBank KC608268 KC614497 F2809 1051 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 Named as Geoplana sp. 4 in GenBank KC608285 KC614511 Geoplaninae 6 F2189 348 Parque Estadual do Desengano, RJ 18 March 2008 −21.87667 −41.91972 Named as Geoplana sp. 5 in GenBank KC608278 KC614505 Geoplaninae 7 F3191 1066 Parque Estadual da Serra do Tabuleiro, SC 9 January 2009 −27.95989 −48.75761 Named as Geoplana sp. 6 in GenBank KC608308 KC614530 F3247 1067 Parque Estadual da Serra do Tabuleiro, SC 13 January 2009 −27.98139 −48.74806 Named as Geoplana sp. 6 in GenBank KC608309 KC614531 Gusana sp. 1 F4421 1088 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608331 KC614549 F4428 1089 Parque Hualpén (Chile) 7 July 2010 −36.79611 −73.15244 GenBank KC608332 – Imbira guaiana (Leal-Zanchet & Carbayo, 2001) F0432 653 Floresta Nacional de São Francisco de Paula, RS 1 August 2004 −29.41666 −50.38333 GenBank KC608229 KC614467 F3361 1071 Floresta Nacional de São Francisco de Paula, RS 20 January 2009 −29.42367 −50.38711 GenBank KC608314 – Imbira marcusi Carbayo et al., 2013 F1041 1190 São Sebastião, SP 8 September 2006 −23.75203 −45.631027 GenBank KC608241 – F2833 404 Parque Nacional Serra da Bocaina, SP September 2008 −22.75222 −44.62194 GenBank KC608291 KC614516 Issoca jandaia Froehlich, 1955 F2043 1015 Parque Nacional Serra da Bocaina, SP 9 February 2007 −22.78222 −44.60583 GenBank KC608270 KC614498 Issoca rezendei (Schirch, 1929) F1182 667 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608248 KC614480 F1679 1010 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608263 KC614493 Issoca sp. 1 F2158 1020 Parque Estadual do Desengano, RJ 16 March 2008 −21.87694 −41.92583 GenBank KC608275 KC614502 F2507 1045 Reserva Biológica Augusto Ruschi, ES 24 May 2008 −19.88740 −40.54319 GenBank KC608279 KC614506 F4085 1085 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608328 KC614546 Luteostriata abundans (Graff, 1899) no voucher São Leopoldo, RS 1998 −29.78828 −51.15084 GenBank KC631621 KC614556 F0205 646 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608223 KC614462 F0238 648 Parobé, RS 27 March 2004 −29.62940 −50.83110 GenBank KC608224 KC614463 Luteostriata ceciliae (Froehlich & Leal-Zanchet, 2003) F3451 1077 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608321 KC614541 Luteostriata ernesti (Leal-Zanchet & Froehlich, 2006) no voucher São Paulo, SP 2005 −23.49080 −46.52090 GenBank – KC614555 F3358 1070 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42367 −50.38711 GenBank KC608313 KC614534 Luteostriata graffi (Leal-Zanchet & Froehlich, 2006) F0384 652 Taquara, RS 28 January 2010 −29.660524 −50.746782 GenBank KC608227 KC614466 F0440 654 Morro das Aranhas, Florianópolis, SC 9 September 2004 −27.468485 −48.377724 GenBank KC608230 KC614468 F3381 1072 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42963 −50.39780 GenBank KC608315 KC614535 Luteostriata muelleri (Diesing, 1861) No voucher GenBank – KC614554 F3268 1199 Parque Estadual da Serra do Tabuleiro, SC 15 January 2009 −27.84278 −48.92583 GenBank KC608311 – Luteostriata sp. F1192 668 Parque da Previdência, São Paulo, SP 9 August 2007 −23.58080 −46.72709 GenBank KC608249 KC614481 Luteostriata sp. 1 F1036 659 São Sebastião, SP 7 September 2006 −23.75203 −45.631027 GenBank KC608239 KC614473 F1078 663 Estação Biológica de Boraceia, Salesópolis, SP, Brazil 2 February 2006 −23.65388 −45.88910 GenBank KC608243 KC614476 Luteostriata sp. 2 F0441 655 Santinhos, Florianópolis, SC 9 September 2004 −27.46370 −48.38100 GenBank KC608231 KC614469 Matuxia matuta (E. M. Froehlich, 1955) F2184 1021 Parque Estadual do Desengano, RJ 17 March 2008 −27.95989 −48.75761 GenBank KC608276 KC614503 F2187 1022 Parque Estadual do Desengano, RJ 17 March 2008 −21.87138 −41.91472 GenBank KC608277 KC614504 Matuxia tuxaua E. M. Froehlich, 1955 F1275 1192 Parque Nacional da Serra dos Órgãos, RJ 16 December 2007 −22.45528 −42.99750 GenBank KC608253 – F3059 1058 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608302 KC614525 Notogynaphallia albonigra (Riester, 1938) F4081 1083 Parque Estadual do Desengano, RJ 13 August 2009 −21.87555 −41.91361 GenBank KC608327 KC614545 Notogynaphallia parca (E. M. Froehlich, 1955) F2972 1196 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.42914 −46.63250 GenBank KC608298 – Notogynaphallia sexstriata (Graff, 1899) F0792 656 Cidade Universitária, USP, São Paulo, SP 19 December 2005 −23.56671 −46.73017 GenBank KC608232 KC614470 F1413 680 Parque Nacional da Serra de Itajaí, SC 4 January 2008 −27.04836 −49.09200 GenBank KC608257 KC614487 Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015 F1399 1007 Parque Nacional da Serra do Itajaí, Blumenau, SC 4 January 2008 −27.05306 −49.08528 GenBank KC608256 KC614486 F1504 698 Parque Nacional da Serra do Itajaí, Blumenau, SC 6 January 2008 −27.05306 −49.08528 GenBank KC608259 KC614489 F1566 707 Parque Nacional da Serra do Itajaí, Blumenau, SC 7 January 2008 −27.05306 −49.08528 GenBank KC608260 KC614490 Obama burmeisteri (Schultze & Müller, 1857) F0989 658 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608234 – F2886 733 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608297 KC614520 Obama carinata (Riester, 1938) F3084 1062 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608304 KC614527 F3093 1064 Parque Estadual Intervales, SP 12 December 2008 −24.27638 −48.41556 GenBank KC608307 KC614529 Obama josefi (Carbayo & Leal-Zanchet, 2001) F3389 1074 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608317 KC614537 F3403 1075 Floresta Nacional de São Francisco de Paula, RS 21 January 2009 −29.42736 −50.39838 GenBank KC608318 KC614538 Obama ladislavii (Graff, 1899) F1418 681 Parque Nacional da Serra de Itajaí, SC 5 January 2008 −27.04836 −49.09200 GenBank KC608258 KC614488 Paraba franciscana (Leal-Zanchet & Carbayo, 2001) F3357 1069 Floresta Nacional de São Francisco de Paula, RS 2 November 2006 −29.42894 −50.39200 GenBank KC608312 KC614533 F5485 935 Floresta Nacional de São Francisco de Paula, RS 12 August 1999 −29.36666 −50.38333 GenBank KC608335 KC614552 Paraba multicolor (Graff, 1899) F1736 594 Parque Nacional Saint Hilaire/Lange, PR 14 January 2008 −25.46972 −48.81222 GenBank KC608265 KC614495 F2997 1055 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608299 KC614521 Paraba rubidolineata (Baptista & Leal-Zanchet, 2005) F5482 936 Floresta Nacional de São Francisco de Paula, RS 26 August 1999 −29.36666 −50.38333 GenBank KC608333 KC614550 Pasipha chimbeva (E. M. Froehlich, 1955) F2813 403 Parque Nacional Serra da Bocaina, SP 8 September 2008 −22.73389 −44.61639 GenBank KC608286 KC614512 F3010 476 Parque Estadual Serra da Cantareira, SP 14 December 2008 −23.42914 −46.63250 GenBank KC608300 KC614522 Pasipha pasipha (Marcus, 1951) F1841 1012 Parque Estadual Alberto Löfgren, São Paulo, SP 26 January 2008 −23.45925 −46.63730 GenBank KC608266 KC614496 F2871 1053 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608295 KC614518 Pasipha pinima (E. M. Froehlich, 1955) F1714 1011 Parque Nacional Saint Hilaire/Lange, PR 13 January 2008 −25.46972 −48.81222 GenBank KC608264 KC614494 F2585 717 Parque Nacional da Serra dos Órgãos, RJ 10 July 2008 −22.45528 −42.99750 GenBank KC608280 KC614507 Pasipha rosea (E. M. Froehlich, 1955) F1061 660 Parque Estoril, São Bernardo do Campo, SP 22 October 2006 −23.77368 −46.51469 GenBank KC608242 KC614475 Pasipha sp. 2 F1138 664 Parque Nacional da Serra dos Órgãos, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608245 KC614477 Pasipha tapetilla (Marcus, 1951) F2878 732 Parque Estadual Serra da Cantareira, SP 26 October 2008 −23.46047 −46.63672 GenBank KC608296 KC614519 F5631B 938 São Paulo, SP May 2007 −29.36666 −50.38333 GenBank KC608336 KC614553 Polycladus sp. 1 F0397 1186 Valdívia (Chile) 2004 −39.81000 −73.23000 GenBank KC608228 – Supramontana irritata Carbayo & Leal-Zanchet, 2003 F3452 772 Floresta Nacional de São Francisco de Paula, RS 25 January 2009 −29.42064 −50.39394 GenBank KC608322 KC614542 F5483 937 Floresta Nacional de São Francisco de Paula, RS 9 February 2006 −29.36666 −50.38333 GenBank KC608334 KC614551 Xerapoa hystrix Froehlich, 1955 F3057 1057 Parque Estadual Serra da Cantareira, SP 12 December 2008 −24.27638 −48.41556 GenBank – KC614524 Xerapoa pseudorhynchodemus (Riester, 1938) F0992 1187 Parque Ecológico do Tietê, São Paulo, SP 30 March 2006 −23.48570 −46.50959 GenBank KC608235 – F1013 1189 Cidade Universitária, USP, São Paulo, SP 17 March 2006 −23.56671 −46.73017 GenBank KC608237 – Xerapoa trina (Marcus, 1951) F1134 1191 Cascata Imbuí, Teresópolis, RJ 14 January 2007 −22.38455 −42.97681 GenBank KC608244 – F1239 669 Parque Nacional da Serra dos Órgãos, RJ 15 December 2007 −22.45528 −42.99750 GenBank KC608250 – *All sequences obtained for this study. All specimens were collected in Brazil, unless specified otherwise. Abbreviations for the Brazilian States are as follows: PR, Paraná; RJ, Rio de Janeiro; RS, Rio Grande do Sul; SC, Santa Catarina; SP, São Paulo. View Large For both datasets and optimality criteria, the genomic regions were concatenated and analysed simultaneously. We also analysed individual partitions (i.e. COI and EF-1-α) to evaluate the information content of each fragment. However, our working topology was based on the simultaneous analyses of both regions. Direct measures of nodal support (sensuGrant & Kluge, 2008a) were accessed in reference to optimality criteria. For MP analyses, nodal support for selected nodes was evaluated using Goodman–Bremer values (GBS; Goodman et al., 1982; Bremer, 1988; Bremer, 1994; see Grant & Kluge, 2008a), calculated with a modified version of the script BREMER.RUN distributed with TNT. This script considered 1000 replicates with ten repetitions of ratchet and drift (Nixon, 1999; Goloboff et al., 2008) in constrained searches and the remaining default parameters. For ML analyses, log-likelihood difference support (LLD; sensuLee & Hugall, 2003; see Meireles et al., 1999; Grant & Kluge, 2008b) was calculated for selected nodes using constrained negative searches in GARLI under the same configuration settings as the initial tree search. Finally, putative transformations for selected branches were compiled using the consensus tree obtained by TNT with YBYRÁ (Machado, 2015). Species recognition Species hypothesis was based on congruence of molecular clades and cohesive morphology. Initially, we recognized all specimens sharing the same set of external and internal traits as morphospecies, as generally used for the taxonomy of the geoplaninids. These morphospecies or putative species were subsequently tested against molecular phylogenies. If morphological and molecular results were congruent with each other for certain morphospecies, we considered the species status as corroborated by molecular and morphological data. By doing so, we are mirroring the phylogenetic concept of species, meaning ‘diagnosable cluster of individual organisms within which there is a parental pattern of ancestry and descent’ (Cracraft, 1983). If the molecular data did not contradict the species hypothesis (usually owing to lack of resolution) but we had morphological evidence support our species hypotheses, we erected the species based on morphological attributes alone. Abbreviations used in figures cg, cyanophil gland cells; co, common glandular ovovitelline duct; e, eye; ej, ejaculatory duct; ep, esophagus; fa, female atrium; fd, female genital duct; g, gonopore; i, intestine; lc, longitudinal cutaneous muscles; lp, longitudinal parenchymatic muscles; m, muscle; ma, male atrium; mo, mouth; ms, thin muscle fibre; o, ovary; ov, ovovitelline duct; pb, penis bulb; ph, pharyngeal pouch; pp, penis papilla; px, pharynx; sb, subintestinal transverse muscles; sd, sperm duct; se, necks of secretory cells; sg, shell glands; sp, supraintestinal transverse muscles; t, testis; vi, vitellaria; vn, ventral nerve plate. MOLECULAR RESULTS We compiled nucleotide sequences of 154 terminals, including five outgroup taxa members of Rhynchodeminae and Geoplaninae individuals (Table 1). These sequences or gene fragments were either obtained from GenBank or they were generated by us. Geoplana was represented by 42 individuals, within which we generated new nucleotide sequences for 37. The remaining ones (five) were obtained from GenBank. The all-terminals dataset consisted of 146 sequences of COI (36 of Geoplana) with 676 bp, and 121 sequences of EF-1-α (28 of Geoplana) with 548 bp. For this dataset, all individuals assigned to Geoplana were represented by both gene fragments, except for G. chita and G. mogi sp. nov. (see below), which were represented by sequences of EF-1-α only, and G. apua sp. nov., represented by COI only. The selected-terminals dataset, for which both gene fragments were available, included a total of 106 individuals of Geoplaninae and five outgroup taxa, within which there were 27 individuals of Geoplana. The parsimony analysis for the all-terminals dataset resulted in 27 equally parsimonious topologies (MPTs) with 5469 steps (Figs 2, 3; see also Supporting Information, Figs S1, S2). The parsimony analysis of the selected-terminals dataset resulted in ten MPTs with 4980 steps. The consensus tree of the selected-terminals dataset (Supporting Information, Fig. S1) recovered similar clades for Geoplana, thus suggesting that missing data did not affect the evidential support for putative species within the genus. Figure 2. View largeDownload slide Summary of sister-group relationships within Geoplaninae based on the simultaneous analysis of COI and EF-1-α partial sequences for all-terminals dataset using parsimony as the optimality criteria with 5469 steps in length. Support values for selected nodes are above branches [Goodman–Bremer (GBS)/log-likelihood difference (LLD)]. Absence of values for LLD indicated that the node was not recovered by the maximum likelihood (ML) analyses. Figure 2. View largeDownload slide Summary of sister-group relationships within Geoplaninae based on the simultaneous analysis of COI and EF-1-α partial sequences for all-terminals dataset using parsimony as the optimality criteria with 5469 steps in length. Support values for selected nodes are above branches [Goodman–Bremer (GBS)/log-likelihood difference (LLD)]. Absence of values for LLD indicated that the node was not recovered by the maximum likelihood (ML) analyses. Figure 3. View largeDownload slide Phylogenetic hypotheses for species of Geoplana based on the simultaneous analysis of COI and EF-1-α partial sequences for all-terminals dataset using parsimony as the optimality criteria. Bases on branches represent unambiguous unique synapomorphies (black) and homoplastic synapomorphies (red) for the respective clades (numbers below bases indicate position of the transformations). Nucleotide positions 1–676 refer to COI, and 677–1225 refer to EF-1-α. Support values for selected nodes are above branches [Goodman–Bremer (GBS)/log-likelihood difference (LLD)]. Absence of values for LLD indicated that the node was not recovered by the maximum likelihood (ML) analyses. *Sequences were obtained for this study. >Species name updated with respect to GenBank. Pictures of specimens are at different scales. Figure 3. View largeDownload slide Phylogenetic hypotheses for species of Geoplana based on the simultaneous analysis of COI and EF-1-α partial sequences for all-terminals dataset using parsimony as the optimality criteria. Bases on branches represent unambiguous unique synapomorphies (black) and homoplastic synapomorphies (red) for the respective clades (numbers below bases indicate position of the transformations). Nucleotide positions 1–676 refer to COI, and 677–1225 refer to EF-1-α. Support values for selected nodes are above branches [Goodman–Bremer (GBS)/log-likelihood difference (LLD)]. Absence of values for LLD indicated that the node was not recovered by the maximum likelihood (ML) analyses. *Sequences were obtained for this study. >Species name updated with respect to GenBank. Pictures of specimens are at different scales. Maximum likelihood analyses considered the TPM1uf+I+Γ model for COI, SYM+I+Γ for EF-1-α, and GTR+I+Γ for the concatenated dataset. Analyses including all terminals assuming a single substitution model for all partitions resulted in a topology with a score of log-likelihood (lnL) −25150.054229, whereas the score for partition models (assuming different substitution models for each fragment) was lnL −24525.1728. The latter partition model was favoured by the AICc (50119.8095 vs. 51476.3247). Analyses including selected terminals assuming a single model for all partitions resulted in a topology with a score of lnL −23020.1415, whereas the score assuming different substitution models for each fragment was lnL −22437.9183. As before, our analyses favoured the the latter (AICc of 45633.2577 vs. 46779.9146). The results of ML analyses are partly congruent with the phylogenetic pattern recovered by the MP analyses (Supporting Information, Figs S2, S3). In general, internal nodes are sensitive to optimality criteria (i.e. MP vs. ML); however, both recovered the clades we recognize as putative species of Geoplana. Phyletic status of Geoplaninae and its main taxonomic groups All phylogenetic analyses revealed that Geoplaninae and five of its genera, namely Cephaloflexa Carbayo & Leal-Zanchet, 2003, Geoplana Schultze & Müller, 1857, Imbira Carbayo et al., 2013, Obama Carbayo et al., 2013 and Matuxia Carbayo et al., 2013, are monophyletic, regardless of the dataset and optimality criterion applied. Two suprageneric assemblages, one constituted by Issoca Froehlich, 1955a + Luteostriata Carbayo, 2010 + Supramontana Carbayo & Leal-Zanchet, 2003 (from Brazil), another by Gusana Froehlich, 1978 + Polycladus Blanchard, 1845 (from Chile), were also recovered in all trees. Choeradoplana Graff, 1896, Issoca, Luteostriata, Paraba Carbayo et al., 2013, Pasipha Ogren & Kawakatsu, 1990 and Supramontana always resulted in polyphyletic assemblages. Sister-group relationships are unstable for these assemblages and for each of these genera (e.g. compare Fig. 2 with Supporting Information, Fig. S3). Species of Geoplana: congruence between molecular and morphological data Most species possess sets of molecular homoplastic synapomorphies, and a few species, namely Geoplana paranapiacaba, Geoplana caraguatatuba, G. pulchella, G. vaginuloides, Geoplana sp. 1 and Geoplana sp. 4, additionally exhibit between one and three unambiguous unique synapomorphies (Fig. 3). That is to say that at least these six species can be diagnosed on a molecular basis. However, we found it premature to include those sets as part of the formal diagnosis of putative species given the nature of most transformations. Congruence between molecular and morphological data of members of Geoplana was retrieved in most analyses. The instances in which phylogenetic analyses were not congruent with our initial concept of morphospecies involved lack of phylogenetic resolution. The phylogenetic analysis of COI (Fig. S5) recovered clades with congruent specimens that share cohesive morphological attributes. However, there were no sequences of this marker for G. chita (see below). Analyses including only EF-1-α recovered most species, but the reciprocal monophyly of Geoplana boraceia, Geoplana cambara, Geoplana cananeia, G. chita and Geoplana ibiuna was not observed because their representatives nested in a polytomy (Supporting Information, Fig. S6). The simultaneous phylogenetic analyses of COI and EF-1-α, independently of taxonomic representation (i.e. all-terminals or selected-terminals datasets) and optimality criteria (i.e. MP and ML) recovered the monophyly of most putative species of Geoplana (see Fig. 3; Supporting Information Figs S1–S6). There was a single exception to this general pattern for G. cambara and G. chita, for which the resolution among haplotypes remained unresolved (Fig. 3), most probably attributable to the lack of COI sequences for G. chita. Four lineages, namely Geoplana sp. 1–4, which are not described here, were also retrieved as distinct monophylic groups; two of these lineages are represented by immature individuals (Supporting Information, Table S1). MORPHOLOGICAL RESULTS Geoplanidae Stimpson, 1857 Geoplaninae Stimpson, 1857 Geoplana Stimpson, 1857 Geoplana vaginuloides (Darwin, 1844) Figs 4and 5 Planaria vaginuloides Darwin, 1844, p. 244. Geoplana vaginuloides: Riester, 1938, p. 72–75 [part]; Froehlich, 1956a, p. 314–315 [part]; Marcus, 1951, p. 54–56 (misidentification), 1952: p. 76–77 (misidentification). Material examined Neotype MZUSP PL 2074 (field number F6387): Parque Estadual da Pedra Branca, Rio de Janeiro/RJ, Brazil (22°56′13.5″S, 043°27′39.5″W). F. Carbayo et al., collected (coll.), 13 December 2014. Cephalic region: horizontal sections on two slides; ovarian region: horizontal sections on eight slides; piece behind ovarian region: horizontal sections on ten slides; pre-pharyngeal region: transverse sections on ten slides; pharynx: sagittal sections on five slides; copulatory apparatus: sagittal sections on ten slides. SMF N574: Barreira, Teresópolis, Rio de Janeiro/RJ, Brazil. E. Bresslau coll., 1914. Cephalic region preserved in balsam on one slide; pre-pharyngeal region: transverse sections on one slide; pharynx: sagittal sections on two slides; copulatory apparatus: sagittal sections on three slides; tail preserved in balsam on one slide. SMF N628: Barreira, Teresópolis, Rio de Janeiro/RJ, Brazil. E. Bresslau, Coll. 1914. Cephalic region and tail preserved in balsam on one slide; pre-pharyngeal region: transverse sections on one slide; pharynx and copulatory apparatus: sagittal sections on three slides. Note We could not confirm conspecificity of some specimens studied by Riester (1938) and specimens examined by Froehlich (1955a, 1956a, 1958) (see Supporting Information, Table S1). Distribution Areas covered with Atlantic forest in the municipalities of Rio de Janeiro and Teresópolis, state of Rio de Janeiro. Diagnosis Dorsal colour pattern constituted by a median black band, bounded on either side by a fine white-yellow stripe, externally to which is a very thin black line, and this is externally bordered by a marginal reddish-iron band. Inner pharynx musculature with two layers. Anteroventral region of penis bulb more developed. Penis papilla with dorsal insertion strongly displaced anteriorly to level the ventral insertion. Type locality Rio de Janeiro, Brazil. External aspect Live animals ~70 mm long and 4 mm wide. Elongated body, with margins approximately parallel; anterior end rounded, posterior pointed. Dorsum convex, ventral side slightly convex. Dorsal colour constituted by a median black band (one-third of body width), bounded on either side by a fine white-yellow stripe (two-ninths), externally to which is a very thin black line (one-ninth), and this is bordered externally by a marginal reddish-iron band (one-third) (Fig. 4A). Ventral body surface grey with dark margins at cephalic end, and saffron yellow on the remaining surface. After fixation, body colour faded slightly. Two types of eyes: (1) a conical type (zuckerhutförmig, sugarloaf shaped), 90 µm high and 40 µm in width; and (2) a cup-shaped type, 40 µm in diameter, with the former contouring the anterior end. Cup-shaped-type eyes spread as a band, one-third of body width on either side. In the posterior half of the body, the eyes even reach to the median black band. Sensory pits simple invaginations, 20 μm deep, located ventromarginally in a single row from the very anterior body tip up to at least the first one-quarter of body length. Relative position mouth-to-body length, 48%. Relative position gonopore- to-body length, 61%. Figure 4. View largeDownload slide Geoplana vaginuloides (Darwin, 1844). Specimen MZUSP PL 2074. A, dorsal view of live specimen, ~60 mm in length. B, photomicrograph of longitudinal and transverse muscles of the subintestinal parenchymatic layer in a horizontal section. C, diagrammatic representation of the pharynx from sagittal sections. D, diagrammatic representation of the copulatory apparatus from sagittal sections. Figure 4. View largeDownload slide Geoplana vaginuloides (Darwin, 1844). Specimen MZUSP PL 2074. A, dorsal view of live specimen, ~60 mm in length. B, photomicrograph of longitudinal and transverse muscles of the subintestinal parenchymatic layer in a horizontal section. C, diagrammatic representation of the pharynx from sagittal sections. D, diagrammatic representation of the copulatory apparatus from sagittal sections. Internal morphology Creeping sole 97% of body width. Very abundant rhabditogen cells piercing dorsal and marginal epidermis. Glandular margin absent. The cutaneous musculature comprises three layers, namely a subepithelial circular layer, followed by a double diagonal layer with decussate fibres, and then a well-developed longitudinal layer. Muscle fibres of the longitudinal, innermost layer (30 µm thick) arranged into bundles with 15–35 fibres each. CMI, 8%. Three parenchymal muscle layers present, all weakly developed: dorsal layer of decussate diagonal fibres, supraintestinal layer of transverse and longitudinal fibres, and subintestinal layer with transverse and longitudinal fibres (Fig. 4B). Ventral nerve plate present. Mouth situated at a distance from the root of the pharynx equivalent to 67% of pharyngeal pocket length. Pharynx cylindrical (Fig. 4C). Oesophagus-to-pharynx ratio, 12%. Outer pharyngeal musculature consisting of a subepithelial layer (6 µm) of longitudinal muscle, followed by a layer (30 µm) of circular fibres. Inner pharynx musculature consisting of a subepithelial layer of circular fibres (40 µm), followed by a layer of of longitudinal fibres (10 µm). Testes dorsally located between supraintestinal parenchymal muscle layer and intestine; anteriormost testes at a distance equivalent to 17% of body length; posterior follicles near to the pharynx root. Penis bulb oval, relatively strongly developed, more developed ventro-anteriorly; it extends from 0.5 mm anterior to penis papilla to being level with the gonopore region (specimen MZUSP PL 2074). Penis bulb consists of tightly packed muscle fibres variously orientated anteriorly, and longitudinally and obliquely orientated posteriorly; in this posterior region, fibres on the right side run obliquely downwards to embrace the ventro-anterior portion of the female atrium and, subsequently, presumably anchor on ventral epidermis. Sperm ducts are dorso-internal to the ovovitelline ducts. Lateral to penis insertion, sperm ducts bend medially and then ventrolaterally to penetrate the bulb. The ducts subsequently run anteriad and join to continue posteriorly and sinuously as the ejaculatory duct (Fig. 4D). Ejaculatory duct traverses central region of penis papilla to open at its tip. Sperm ducts lined with a cuboidal, ciliated epithelium and surrounded by a 5-µm-thick layer of circular muscles. Wall of ejaculatory duct stained reddish, consisting of a cuboidal, ciliated epithelium, surrounded by circular muscles, which constitute a muscular cylinder with a diameter equivalent to three times the diameter of the duct (Fig. 5A). Figure 5. View largeDownload slide Geoplana vaginuloides (Darwin, 1844). A, photomicrograph of part of the penis papilla of specimen N574 on a sagittal section, showing the ejaculatory duct and its surrounding muscle. B, photomicrograph of the female genital duct of specimen MZUSP PL 2074 on a sagittal section. Figure 5. View largeDownload slide Geoplana vaginuloides (Darwin, 1844). A, photomicrograph of part of the penis papilla of specimen N574 on a sagittal section, showing the ejaculatory duct and its surrounding muscle. B, photomicrograph of the female genital duct of specimen MZUSP PL 2074 on a sagittal section. Penis papilla very long, 5 mm, with dorsal and ventral insertions at the same horizontal plane (Fig. 4D). This papilla is cylindrical along most of its length; total length equal to 17 times its diameter, entirely occupying both male and female atria; dorsal insertion strongly displaced anteriorly and very ventrally, so that it projects from the anteroventral portion of the male atrium. Subepithelial musculature consisting of a dense, 42-µm-thick layer of circular muscle, tending to decussate distally, followed by a single layer of longitudinal muscle. Epithelium of papilla pierced by reddish secretions. Poor condition of sections does not display further histological details. Male atrium long and ample, not folded. Ovaries oval shaped, 250 µm in maximal length, and situated at a distance from anterior end equivalent to one-fifth of body length. Ovovitelline ducts emerge from an unknown region of ovaries, lateral to female atrium. Then, they curve medially and join to form the common glandular ovovitelline duct, dorsally to the posterior third of this atrium (Fig. 4D). Common glandular ovovitelline duct as long as one-third of female atrium length and communicates with the female genital duct; this duct is a projection of the posterior region of the female atrium and is dorso-anteriorly orientated. The female genital duct is lined with an epithelium provided with intraepithelial gaps (Fig. 5B). Poor condition of sections does not display further histological details. Female atrium without folds, two to three times as long as male atrium, lined with a tall epithelium, which posteriorly presents intraepithelial gaps. Atrial epithelium surrounded by a thin layer of longitudinal muscle fibres, followed by a layer of muscles with criss-cross arrangement; thickness of both layers equal to 25 µm, distally doubling in thickness. Remarks The original description of the dorsal chromatic pattern of this species may be ambiguous (“Sides and foot coloured dirty ‘orpiment orange’; above with two stripes on each side of pale ‘primrose-yellow’, edged externally with black; on centre of the back a stripe of glossy black”; Darwin, 1844, p. 244). From Darwin’s description, Marcus deduced that ‘an additional black lateral stripe should exist on each side, albeit only the external stripe was mentioned’ (Marcus, 1952 [original in Portuguese]). No specimen matching Marcus’ description was ever found, but this ambiguity is removed in Darwin’s field notes. In an entry on 17 June, 1832, he states: “Colours: back with glossy black stripe; on each side of this a primrose white one edged externally with black; these stripes reach to extremities, & become uniformly narrower. sides & foot dirty ‘orpiment orange’” (in Keynes, 2000, p. 47). We found this precise colour pattern displayed by the specimen MZUSP PL 2074 from the type locality. Hence, it is most probable that this specimen and that found by Darwin are conspecific. A conclusive comparison is not possible because there is no record of Darwin’s specimen in the Natural History Museum, London, where the material collected by Darwin is deposited (H. Jones, personal communication). Neotype designation satisfies provisions of the ICZN code (Art. 75.3.1.–7.) as follows: (1) the neotype is designated with the express purpose of clarifying the taxonomic status or the type locality of the name Geoplana vaginuloides; (2) the characters differentiating G. vaginuloides from any other taxa are stated in the diagnosis above; (3) these data and the description of the species are sufficient to ensure recognition of the specimen designated; (4) after Dr Hugh Jones [from Natural History Museum London (NHM)], the name-bearing type specimen is lost or even has never been deposited in the NHM; (5) the ‘Remarks’ section shows that neotype is consistent with the former name-bearing type from the original description and from Darwin’s field notes (in Keynes, 2000); (6) the neotype came from the original type locality; and (7) the neotype is the property of the Museu de Zoologia da Universidde de São Paulo (MZUSP), Brazil. Geoplana apua Almeida & Carbayo sp. nov. Figs 6and 7 Geoplana vaginuloides: Riester, 1938, p. 72–75 [part]: Froehlich, 1956a, p. 314–315 [part]. Material examined Holotype MZUSP PL 2072 (field number F5236): Nova Iguaçú/RJ, Brazil. Cristiano Sampaio et al., coll., 4–6 April 2012. Cephalic region: horizontal sections on eight slides; ovarian region: sagittal sections on five slides; pre-pharyngeal region: transverse sections on ten slides; pharynx: sagittal sections on six slides; copulatory apparatus: sagittal sections on 11 slides. Paratype MZUSP PL 2090 (field number F7001): Teresópolis, Rio de Janeiro/RJ, Brazil. E. M. Froehlich coll., 20 July 1952. Copulatory apparatus: sagittal sections on 18 slides. Paratype SMF N460: Barreira, Teresópolis, Rio de Janeiro/RJ, Brazil. Ernest Bresslau coll., 25 February 1914. Cephalic region and tail preserved in balsam on one slide; pre-pharyngeal region: transverse sections on one slide; pharynx and copulatory apparatus: sagittal sections on two slides. Distribution Areas covered with Atlantic forest in the municipalities of Nova Iguaçú and Teresópolis, State of Rio de Janeiro, Brazil. Diagnosis Dorsal colour pattern constituted by a median luminous orange band, bounded on either side by a black stripe, externally to which is a greenish-yellow stripe, and this again is bordered externally by a marginal black band. Inner pharynx musculature with two layers. Anterodorsal region of penis bulb more developed. Penis papilla with dorsal insertion strongly displaced anteriorly and very ventrally, as long as 18 times its diameter. Etymology The specific epithet is a Tupi (indigenous Brazilian tribe) word that means mound (Tibiriçá, 1984), referring to the hilly terrain where the species was collected. External aspect Live animals 45 mm long when creeping. Body margins parallel; anterior end rounded, posterior end pointed. In preserved specimens, dorsum convex, ventral side slightly convex. Dorsal colour pattern of the preserved specimen F5236 constituted by a median luminous orange band (one-third of body width), bounded on either side by a black stripe (1/12th), externally to which is a greenish-yellow stripe (1/12th), and this again is bordered externally by a marginal black band (one-sixth) (Fig. 6A). Ventral body surface pale orange. In specimen F7001 (preserved for > 60 years), black became brownish, remaining colours became cream. Two types of eyes: a conical type, 80 µm high and 30 µm wide; and a cup-shaped type, 50 µm in diameter. The former type of eye contours the anterior portion of the body. Four millimetres behind anterior end, cup-shaped-type eyes spread dorsally onto the yellowish stripes until rear end of body, where the eyes are sparsely distributed. Sensory pits simple invaginations, 25 μm deep, located ventromarginally in a single row and extending from the very anterior end up to at least the first one-fifth of body length. Relative position mouth-to-body length, 55%. Relative position gonopore-to-body length, 67%. Figure 6. View largeDownload slide Geoplana apua sp. nov. A, dorsal view of the MZUSP PL 2072 preserved specimen on millimetre graph paper after cutting off a piece of the body. B, photomicrograph of longitudinal and transverse muscles of the subintestinal parenchymatic layer in a horizontal section of specimen MZUSP PL 2072. C, diagramatic reconstruction of half of a transverse section of the pre-pharyngeal region of the holotype; dots represent longitudinal parenchymatic muscle fibres. D, photomicrograph of the two muscle layers of the internal musculature of the pharynx in sagittal section. E, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2072. F, photomicrograph of the ejaculatory duct and its surrounding muscle of specimen MZUSP PL 2072 in sagittal section. Figure 6. View largeDownload slide Geoplana apua sp. nov. A, dorsal view of the MZUSP PL 2072 preserved specimen on millimetre graph paper after cutting off a piece of the body. B, photomicrograph of longitudinal and transverse muscles of the subintestinal parenchymatic layer in a horizontal section of specimen MZUSP PL 2072. C, diagramatic reconstruction of half of a transverse section of the pre-pharyngeal region of the holotype; dots represent longitudinal parenchymatic muscle fibres. D, photomicrograph of the two muscle layers of the internal musculature of the pharynx in sagittal section. E, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2072. F, photomicrograph of the ejaculatory duct and its surrounding muscle of specimen MZUSP PL 2072 in sagittal section. Internal morphology Creeping sole 77% of body width. Glandular margin absent. Cutaneous musculature same as in the type species of the genus in terms of organization. CMI, 7%. Muscle fibres of the longitudinal layer (15–20 µm thick) arranged into bundles with ten to 30 fibres each. Parenchymal musculature same as in the type species of the genus in terms of organization and relative development (Fig. 6B). Ventral nerve plate present. Mouth situated at a distance from root of the pharynx equivalent to 54% of pharyngeal pocket length. Pharynx cylindrical (Fig. 6E). Oesophagus-to-pharynx ratio, 18%. Pharyngeal musculature (Fig. 6D) same as in the type species of the genus in terms of organization and development, with the exception of the inner longitudinal layer, which reaches 35 µm in thickness. Testes dorsally located between supraintestinal parenchymal muscle layer and intestine; anteriormost testes at a distance from anterior end equivalent to 13% of body length; posteriormost near the pharynx root. Penis bulb oval shaped, relatively strongly developed, consisting of tightly packed muscle fibres variously orientated. The bulb is more developed dorso-anteriorly, and extends from 0.5 mm anterior to penis papilla up to the gonopore region (specimen MZUSP PL 2072). Diagonal fibres surrounding distal section of penis bulb run obliquely downwards on the right side of the body to embrace ventrally the anterior portion of the female atrium. Sperm ducts run laterally to the ovovitelline ducts. Laterally to ventral insertion of penis papilla, sperm ducts bend anteriorly and medially, then penetrate penis bulb ventrolaterally, and maintain this course until they join each other. From this point onwards, the ejaculatory duct runs posteriorly, centrally traversing the penis papilla to open at its tip. Sperm present in extrabulbar portion of sperm ducts; in one specimen, sperm was also present at the anterior section of the intrabulbar lumen. Sperm ducts lined with a cuboidal, ciliated epithelium and surrounded by a 5- to 8-µm-thick layer of circular muscles. Proximal half of ejaculatory duct lined with cuboidal, ciliated epithelium and pierced by gland cells producing weakly erythrophil, fine granules; distal half lined with squamous, non-ciliated epithelium pierced by gland cells producing erythrophil, coarse granules. Anterior half of this duct surrounded by circular muscles, which constitutes a muscular cylinder in the distal portion with a diameter equivalent to five times the diameter of this duct. Penis papilla thin, somewhat conical, because it narrows very gradually, and very long; total length equal to 18 times its diameter, occupying almost entirely both male and female atria; dorsal insertion strongly displaced anteriorly and very ventrally, so that it projects from anteroventral portion of male atrium. Squamous lining epithelium of the penis papilla pierced by numerous gland cells producing xanthophyl and erythrophil fine granules, and underlain by a dense 85-µm-thick layer of circular muscle, distally tending to decussate, followed by a single layer of longitudinal muscle (Fig. 6F). Male atrium not folded (Fig. 7A, B), lined with a low, non-ciliated epithelium, and underlain by a 5-µm-thick layer of circular muscles, followed by an equally thin layer of longitudinal fibres. A set of muscle fibres is also present as longitudinal fibres extending around the gonopore canal laterally to the male atrium, and being intermingled with those of the penis bulb. Ovaries oval shaped, 300 µm in maximal diameter, and situated at a distance from anterior end equivalent to 22.5% of body length. Ovovitelline ducts emerge from the dorsolateral aspect of ovaries. The oviducts run laterally to the female atrium, curve medially and then join to form the common glandular ovovitelline duct, dorsally to this atrium. Common glandular ovovitelline duct as long as 40% of female atrium length, communicating with female genital duct (Fig. 7B); this duct is a projection of the posterior region of the female atrium, bent dorsally and lined with a squamous, non-ciliated epithelium. A cyanophil fibrillar mass, 5–10 µm thick, is immediately present underneath this epithelium of the female atrium, with very thin circular filaments (< 1 µm) resembling muscle fibres in stain affinity and orientation, followed by a layer of longitudinal muscle fibres. Figure 7. View largeDownload slide Geoplana apua sp. nov. A, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2072. B, diagrammatic representation of the copulatory apparatus of specimen MZUSP PL 2072 from sagittal sections. Figure 7. View largeDownload slide Geoplana apua sp. nov. A, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2072. B, diagrammatic representation of the copulatory apparatus of specimen MZUSP PL 2072 from sagittal sections. Female atrium without folds, as long as three times that of male atrium, and lined with a cuboidal-to-columnar epithelium, which is pierced by glands producing xanthophil granules and is underlain by a thin layer of muscle fibres in a criss-crossed arrangement, followed by a loose layer (25 µm) of longitudinal fibres. Geoplana mogi Almeida & Carbayo sp. nov. Figs 8and 9 Geoplana vaginuloides: Marcus, 1951, p. 54–56. Material examined Holotype MZUSP PL 2089 (field number F7000): Parque do Estado, São Paulo/SP, Brazil. O. Françoso and E. M. Froehlich, coll., 29 May 1985. Cephalic region: sagittal sections on ten slides; ovarian region: horizontal sections on 12 slides; pre-pharyngeal region: transverse sections on ten slides; pharynx: sagittal sections on 20 slides; copulatory apparatus: sagittal sections on 35 slides. Paratype Marcus’ specimen (1951) (from EMF collection): Eldorado neighborhood, West zone of the city of São Paulo/SP, Brazil. E. Marcus and E. M. Froehlich, coll., 1 November 1947. Copulatory apparatus: sagittal sections on five slides. Paratype MZUSP PL 2061 (field number F1022): Mogi das Cruzes/SP, Brazil (23°27′05.5″S, 046°13′40.8″W). F. Carbayo, coll., 27 May 2006. Cephalic region: transverse sections on 13 slides; ovarian region: horizontal sections on seven slides; pre-pharyngeal region: transverse sections on five slides; pharynx: sagittal sections on eight slides; copulatory apparatus: sagittal sections on five slides. Distribution Areas covered with secondary Atlantic forest in the municipalities of São Paulo and Mogi das Cruzes, state of São Paulo, Brazil. Etymology The specific epithet refers to the Tupi-Guarani (indigenous Brazilian tribe) name of the municipality where the species was collected. Diagnosis Dorsal colour pattern constituted by a median orange-ochre band, bounded on either side by a thin whitish stripe, and in turn this is bordered externally by a thin black line, externally to which is a grey band with dense white mottling, externally to which is a thin marginal black line. Longitudinal parenchymal muscle fibres disposed around intestine as a loose muscular tube. Diameter of muscular cylinder surrounding ejaculatory duct equivalent to 1.3 times the diameter of this duct. External aspect Preserved animals 50 mm long and 4 mm wide. Body margins parallel; anterior end rounded, posterior pointed. Dorsum strongly convex, ventral side flat. On histological slides, body 1.25 mm in height. In live animals, dorsal colour pattern constituted by a median orange-ochre band (one-third of body width) that passes into the black of the cephalic region (one-fifth of body width), bounded on either side by a thin whitish stripe (one-ninth), and in turn this is bordered externally by a thin black line, externally to which is a grey band (the two latter totalling two-ninths) with dense white mottling, externally to which is a thin marginal black line (Fig. 8A). This marginal band was discovered in a detailed examination of the preserved specimen; this band remained hidden in dorsal view owing to the strong convexity of the dorsum. Ventral body surface whitish, with blackish margins. In preserved specimen F1022, whitish, blackish and greyish colours lost brightness; orange-ochre faded. In specimen F7000, median band was ochre-brown bordered on either side by cream lines, in turn this was bordered by a brown green line, externally to which was a beige grey band. Two types of eyes: a conical type, 90 µm high and 40 µm in width; and a cup-shaped type, 40 µm in diameter. The former type contours the anterior tip of the body. Behind the first millimetre, cup-type eyes spread progressively onto the dorsum, reaching the whole dorsum from the pharynx region until posterior end. Sensory pits simple invaginations, 55 μm deep, located ventromarginally in a single row, at least along first third of body from very anterior end. Relative position mouth-to-body length, 55%. Relative position gonopore-to-body length, 71%. Figure 8. View largeDownload slide Geoplana mogi sp. nov. A, dorsal view of the live specimen MZUSP PL 2061. B, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2089. C, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2089. Figure 8. View largeDownload slide Geoplana mogi sp. nov. A, dorsal view of the live specimen MZUSP PL 2061. B, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2089. C, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2089. Internal morphology Creeping sole 78% of body width. Glandular margin absent. Three typical geoplaninid cutaneous muscle layers present, being as thick as 9.5% of body height. Muscle fibres of the longitudinal, innermost layer (40 µm thick) arranged into bundles with 18–20 fibres each. Three parenchymal muscle layers present: dorsal layer of decussate diagonal fibres, supraintestinal layer of transverse fibres, and subintestinal layer with transverse fibres. Longitudinal fibres around intestine present, constituting a loose tube with fibres mixing with transverse ones. Ventral nerve plate present. Mouth situated at a distance from root of the pharynx equivalent to 44–46% of pharyngeal pocket length. Pharynx cylindrical, with dorsal insertion slightly posterior to ventral one (Fig. 8B). Oesophagus-to-pharynx ratio, 15%. Outer pharyngeal musculature consisting of a subepithelial layer (5 µm) of longitudinal muscle, followed by a layer (70 µm) of circular fibres, the innermost of which are intermingled with longitudinal muscle fibres. Inner pharynx musculature consisting of a thin subepithelial layer of longitudinal muscle, followed by a layer (70 µm) of circular fibres and an innermost layer (30 µm) of longitudinal fibres. Testes dorsally located between supraintestinal parenchymal muscle layer and intestine; anteriormost testes at a distance from anterior end equivalent to 22% of length of body; posteriormost follicles near pharynx root. Penis bulb elongated, extending posteriorly from 0.5 mm (specimen MZUSP PL 2061) anterior to penis papilla to the level of the gonopore. The bulb consists of tightly packed muscle fibres, variously orientated. Diagonal fibres on the right side of the bulb run obliquely downwards to embrace ventrally the anterior portion of female atrium. These fibres apparently cross the nerve plate to anchor on the ventral epidermis. Sperm ducts run dorsally to the ovovitelline ducts. Laterally to penis insertion, sperm ducts bend medially and then penetrate the penis bulb ventrolaterally. Subsequently, the ducts recurve and separately open into the sinuous ejaculatory duct (Figs 8C, 9A). The latter traverses the central region of the penis papilla to open at its tip. Sperm present in sperm ducts, also along its course within penis bulb. Sperm ducts lined with a cuboidal, ciliated epithelium and surrounded by a 20-µm-thick layer of circular muscles. Ejaculatory duct lined with columnar, ciliated epithelium; epithelium of extrapenial portion pierced by glands producing weakly erythrophil, fine granules, and being surrounded by 60-µm-thick layer of circular muscle fibres. Epithelium of intrapenial portion of ejaculatory duct richly pierced by glands producing erythrophil granules and surrounded by a 20-µm-thick layer of circular muscles, which constitutes a muscular cylinder possessing a diameter equivalent to 1.3 times the diameter of this duct (Fig. 9B). Figure 9. View largeDownload slide Geoplana mogi sp. nov. A, diagrammatic representation of the copulatory apparatus of the specimen MZUSP PL 2089 from sagittal sections. B, photomicrograph of the ejaculatory duct and its surrounding musculature of specimen MZUSP PL 2089 in sagittal section. C, photomicrograph of dorsal epithelium of penis papilla and erythrophil glands located immediately beneath epithelium of the unlabelled specimen studied by Marcus (1951) in sagittal section. Figure 9. View largeDownload slide Geoplana mogi sp. nov. A, diagrammatic representation of the copulatory apparatus of the specimen MZUSP PL 2089 from sagittal sections. B, photomicrograph of the ejaculatory duct and its surrounding musculature of specimen MZUSP PL 2089 in sagittal section. C, photomicrograph of dorsal epithelium of penis papilla and erythrophil glands located immediately beneath epithelium of the unlabelled specimen studied by Marcus (1951) in sagittal section. Penis papilla long and cylindrical along most of its length, with dorsal insertion slightly anterior to the ventral. Total length of papilla equal to ten times its greatest diameter, and occupying male atrium and almost entire female atrium; lined with a cuboidal, non-ciliated epithelium, pierced by two types of gland cells, producing coarse erythrophil granules and fine cyanophil granules, respectively. Erythrophil glands pierce anterior and distal third of penis; necks of cyanophil glands pierce dorsal epithelium of second third, and sometimes also the ventral one, and pack space immediately beneath epithelium so that subepithelial muscle fibres are apart from it (Fig. 9C). Subepithelial musculature of penis papilla consisting of dense 80- to 90-µm-thick layer of circular muscle, the fibres of which are decussate distally, followed by a 10- to 20-µm thick layer of longitudinal muscle. Male atrium not folded, lined with a low, non-ciliated epithelium, pierced by some glands secreting amorphous substance, and underlain by 7-µm-thick layer of circular muscles, followed by a nearly inconspicuous layer of longitudinal fibres. Ovaries elongated, 1.1 mm in maximal anteroposterior diameter, 0.2 mm in dorsoventral width, and situated at a distance from anterior end equivalent to 22% of body length. Ovovitelline ducts emerge from dorsolateral aspect of ovaries laterally to female atrium, curve medially and join common glandular ovovitelline duct dorsally to this atrium. Common glandular ovovitelline duct as long as a third of female atrium length; it communicates with female genital duct, which is a dorso-anteriorly orientated projection of the posterior region of the female atrium (Fig. 9A) and is lined with a 25-µm-tall cuboidal, ciliated epithelium. Female atrium without folds, posterior portion funnel shaped; as long as 1.5 times that of male atrium; anteriorly lined with a cuboidal epithelium; posteriorly lined with a tall epithelium with stratified appearance and intraepithelial holes. Epithelium pierced by two types of glands, producing erythrophil granules and, less abundantly, xanthophil. Epithelium underlain by 5- to 45-µm-thick layer of circular-to-decussate muscle fibres, followed by some longitudinal fibres, and then a layer (50–150 µm thick) of thicker longitudinal fibres. Geoplana piratininga Almeida & Carbayo sp. nov. Figs 10and 11 Geoplana vaginuloides: Marcus, 1952, p. 76–77. Material examined Holotype MZUSP PL 2071 (field number F5134): Parque Jardim Previdência, São Paulo/SP, Brazil (23°34′48.7″S, 046°43′39.3″W). F. Carbayo et al., coll., 5 February 2006. Cephalic region: horizontal sections on four slides; ovarian region: horizontal sections on two slides; region behind ovaries: horizontal sections on ten slides; pharynx: sagittal sections on six slides; copulatory apparatus: sagittal sections on eight slides. Paratype MZUSP PL 2060 (field number F0509): Parque Ecológico do Tietê, São Paulo/SP, Brazil (23°29′27.1″S, 046°31′01.6″W). F. Carbayo, coll., 27 May 2005. Pharynx and post-pharynx region: sagittal sections on six slides (incompletely mature). Paratype MZUSP PL 666 (field number F1163): Parque Jardim Previdência, São Paulo/SP, Brazil (23°34′46.8″S, 046°43′39.0″W). O. M. Junior, coll., 17 June 2007. Pharynx: sagittal sections on 13 slides; copulatory apparatus: sagittal sections on seven slides (incompletely mature). Distribution Areas covered with secondary Atlantic forest and garden in the municipalities of São Paulo and Mongaguá, state of São Paulo, Brazil. Etymology The specific epithet is a Tupi (indigenous Brazilian tribe) word formerly referring to the city of São Paulo, where the species was collected. Diagnosis Dorsal colour pattern constituted by a median traffic red band, bounded on either side by a black stripe, externally to which is a white stripe, and in turn this is externally bordered by a black band, externally to which is a thin marginal white line. Dorsal and ventral insertions of penis papilla at the same transverse plane. Wall of female atrium not folded. Female-to-male atrial length ratio, 2.0. External aspect Live animals 25 mm long and 4 mm wide. Body relatively short, with margins nearly parallel; anterior end rounded, posterior pointed. Dorsum strongly convex, ventral side slightly convex. Dorsal body surface constituted by a median traffic red band (1/3.5th of body width), passing into black part of cephalic region (one-sixth of body length) and of posterior tip, bounded on either side by a black stripe, externally to which is a white stripe (one-seventh), and in turn this is bordered externally by a black band (one-seventh), externally to which is a thin marginal white line (Fig. 10A). Ventral side whitish. After fixation, dorsal black colour faded, red passed into yellow-orange and white into cream. Two types of eyes: a conical type, contouring only anterior end; and a cup-shaped type, spreading dorsally onto the black bands. Sensory pits simple invaginations, 25 μm deep, located ventromarginally in a single row from the very anterior end up to at least first one-fifth of body length. Relative position mouth-to-body length, 42.5%. Relative position gonopore- to-body length, 64%. Figure 10. View largeDownload slide Geoplana piratininga sp. nov. A, dorsal view of the live specimen MZUSP PL 2060; anterior to the left, ~32 mm in length. B, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2071. C, photomicrograph of the three muscle layers of the internal musculature of the pharynx in sagittal section of specimen MZUSP PL 2071. D, photomicrograph of dorsal epithelium of penis papilla and erythrophil glands located immediately beneath the epithelium of specimen MZUP PL 2071 in sagittal section. Figure 10. View largeDownload slide Geoplana piratininga sp. nov. A, dorsal view of the live specimen MZUSP PL 2060; anterior to the left, ~32 mm in length. B, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2071. C, photomicrograph of the three muscle layers of the internal musculature of the pharynx in sagittal section of specimen MZUSP PL 2071. D, photomicrograph of dorsal epithelium of penis papilla and erythrophil glands located immediately beneath the epithelium of specimen MZUP PL 2071 in sagittal section. Internal morphology Creeping sole ~85% of body width, as measured on horizontal or sagittal sections. Glandular margin absent. Three typical geoplaninid cutaneous muscle layers present, being as thick as 5% of body height. Muscle fibres of the longitudinal, innermost layer (25 µm thick) arranged into bundles. Three parenchymal muscle layers present, all weakly developed: dorsal layer of decussate diagonal fibres, supraintestinal layer of transverse and longitudinal fibres, and subintestinal layer with transverse fibres. Additionally, longitudinal fibres surround intestine as a loose tube, the ventral fibres of which are mixed up with those of the subintestinal layer. Ventral nerve plate present. Mouth situated at a distance from root of the pharynx equivalent to 50–65% of pharyngeal pocket length. Pharynx cylindrical, with dorsal insertion slightly posterior to ventral one (Fig. 10B). Oesophagus-to-pharynx ratio, 12%. Outer pharyngeal musculature consisting of a subepithelial layer (5 µm) of longitudinal muscle, followed by a layer (45 µm) of circular fibres, the innermost of which are intermingled with longitudinal ones. Inner pharynx musculature consisting of a thin subepithelial layer of longitudinal muscle, followed by a layer (40 µm) of circular fibres and an innermost layer (10 µm) of longitudinal fibres (Fig. 10C). Testes dorsally located between supraintestinal parenchymal muscle layer and intestine; anteriormost testes at a distance from anterior end equivalent to 25.5% of body length; posteriormost follicles a little anterior to pharynx root. Penis bulb elongated, somewhat oval, extending from 0.3 mm anterior to penis papilla; from thereon the bulb extends ventrally, whereas dorsally it extends posteriorly for only short distance. The bulb consists of variously orientated packed muscle fibres. Diagonal fibres on the right side of the bulb run obliquely downwards to embrace ventrally the anterior portion of the female atrium. These fibres apparently mix with the subintestinal parenchymal musculature, but their anchor point could not be discerned. Sperm ducts run dorsally to the ovovitelline ducts. Sperm ducts bend medially behind insertions of penis papilla and penetrate penis bulb ventrolaterally. The ducts subsequently recurve to join and thus form the ejaculatory duct, which sinuously posteriad, then traverses the penis papilla to open at its tip. Distal portion of sperm ducts containing sperm, along their course both outside and inside penis bulb. Sperm ducts lined with a cuboidal, ciliated epithelium and surrounded by a 6-µm-thick layer of circular muscles. Extrapapillar portion and anterior portion of intrapapillar section of ejaculatory duct lined with columnar ciliated epithelium, pierced by glands producing erythrophil fine granules. Remaining section of the ejaculatory duct also columnar and ciliated, pierced by numerous gland cells producing erythrophil granules. Entire ejaculatory duct surrounded by a layer of circular muscle, 35 µm thick around anterior portion, otherwise 10 µm thick (Fig. 10D). Diameter of muscular cylinder surrounding ejaculatory duct is equivalent of 3.5 times the diameter of this duct. Penis papilla is long, horizontal and cylindrical along most of its length, with dorsal and ventral insertions at the same transverse plane (Fig. 11A, B). Total length of papilla equal to five times its greatest diameter, and completely filling both male and female atrium; papilla lined with a cuboidal epithelium, pierced by gland cells producing coarse erythrophil granules; glands producing cyanophil granules packed in space immediately beneath epithelium of the dorsal second third of penis papilla, so that subepithelial muscle fibres are separated from it (Fig. 10D). Penial subepithelial musculature consisting of dense 90-µm-thick layer of circular muscle, followed by a layer (25 µm thick) of longitudinal muscle. Figure 11. View largeDownload slide Geoplana piratininga sp. nov. A, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2071. B, diagrammatic representation of the copulatory apparatus of specimen MZUSP PL 2071 from sagittal sections. Figure 11. View largeDownload slide Geoplana piratininga sp. nov. A, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2071. B, diagrammatic representation of the copulatory apparatus of specimen MZUSP PL 2071 from sagittal sections. Male atrium ample, not folded, lined with a low, non-ciliated epithelium, pierced by glands secreting erythrophil amorphous substance and underlain by a 15-µm-thick layer of circular muscles, followed by an inconspicuous layer of longitudinal fibres. Ovaries incompletely developed, 500 µm in length along anteroposterior body axis, located above ventral nerve plate and situated at a distance from anterior end equivalent to ~20.5% of body length. Laterally to the female atrium, the oviducts curve medially and join to form a common glandular ovovitelline duct. This duct is as long as one-third of the female atrium length and communicates with the female genital duct, the latter being a funnel-shaped, anterodorsally orientated projection of the posterior region of the female atrium. Female atrium funnel shaped and without folds, as long as two times the male atrium length (Fig. 11B); in its anterior portion it is lined with cuboidal epithelium, posteriorly with a 40-µm-high epithelium with stratified aspect; pierced by two types of glands producing cyanophil and xanthophil granules, respectively. Epithelium underlain by a 5- to 40-µm-thick layer of circular-to-decussate muscle fibres, followed by a 25-µm-thick layer of longitudinal fibres. Additionally, 4-µm-thick longitudinal fibres around these muscles constitute a kind of muscle coat. Geoplana paranapiacaba Almeida & Carbayo sp. nov. Figs 12and 13 Material examined Holotype MZUSP PL 2079 (field number F6660): Cidade Turística de Paranapiacaba, Santo André/SP, Brazil (23°46′38.9″S, 046°18′05.2″W). F. Carbayo et al., coll., 5 June 2015. Cephalic region: horizontal sections on four slides; ovarian region: horizontal sections on nine slides; region behind ovaries: horizontal sections on 14 slides; pre-pharyngeal region: transverse sections on 16 slides; pharynx: sagittal sections on 17 slides; copulatory apparatus: sagittal sections on 20 slides. Paratype MZUSP PL 2087 (field number F6998): Cidade Turística de Paranapiacaba, Santo André/SP, Brazil. E. M. Froehlich et al., coll., 13 May 1990. Copulatory apparatus: sagittal sections on 39 slides. Distribution Only known from the type locality. Etymology The specific epithet refers to the Tupi-Guarani (indigenous Brazilian tribe) name of the district where the type material was collected. Diagnosis Dorsal colour pattern constituted by a median stripe of orange-ochre, bounded on either side by a black line, externally to which is a broader white line, and in turn this is externally bordered by a thin black line, externally to which is a grey band with dense white mottling, bordered by a thin marginal black line. Diameter of muscular cylinder round ejaculatory duct equivalent to 0.5 times the diameter of this duct. Female-to-male atrial length ratio, 3.5–4. External aspect Preserved animals up to 50 mm long, and 4 mm wide. Body margins nearly parallel; anterior end rounded, posterior end pointed. Dorsum strongly convex, ventral side slightly convex. Dorsal body surface constituted by a median stripe of orange-ochre (one-quarter of body width) that passes into the black of cephalic region (one-fifth of body length), bounded on either side by a black line (1/16th) posteriorly merging with each other, externally to which is a broader white line (one-eighth), and this is externally bordered by a black stripe (1/16th), externally to which is a grey stripe (one-eighth) with dense white mottling, then bordered by a marginal thin black line (Fig. 12A). Ventral body surface is whitish with blackish margins. In preserved specimen F6660, dorsal colour slightly faded. In preserved specimen F6998, stripes were varied from faded black grey to pearl orange. Two types of eyes: a conical type contouring only anterior end; and a cup-shaped type, spreading dorsally onto the black stripes. Sensory pits simple invaginations, 40 μm deep, located ventromarginally in a single row from the very anterior tip to at least the end of the anterior fifth of the body. Relative position mouth-to-body length, 61%. Relative position gonopore-to-body length, 74%. Figure 12. View largeDownload slide Geoplana paranapiacaba Almeida & Carbayo sp. nov. A, dorsal view of the 50-mm-long specimen MZUSP PL 2079, immediately after having been killed with boiling water. It was accidentally cut into two pieces at the moment of collection. B, photomicrograph of longitudinal and transverse muscles of the subintestinal parenchymatic layer in horizontal section of specimen MZUSP PL 2079. C, diagramatic reconstruction of half of a transverse section of the pre-pharyngeal region of the holotype; dots represent longitudinal parenchymatic muscle fibres. D, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2079. E, photomicrograph of dorsal epithelium of penis papilla and erythrophil glands located immediately beneath the epithelium of specimen MZUP PL 2079 in sagittal section. Figure 12. View largeDownload slide Geoplana paranapiacaba Almeida & Carbayo sp. nov. A, dorsal view of the 50-mm-long specimen MZUSP PL 2079, immediately after having been killed with boiling water. It was accidentally cut into two pieces at the moment of collection. B, photomicrograph of longitudinal and transverse muscles of the subintestinal parenchymatic layer in horizontal section of specimen MZUSP PL 2079. C, diagramatic reconstruction of half of a transverse section of the pre-pharyngeal region of the holotype; dots represent longitudinal parenchymatic muscle fibres. D, photomicrograph of a sagittal section of the pharynx of specimen MZUSP PL 2079. E, photomicrograph of dorsal epithelium of penis papilla and erythrophil glands located immediately beneath the epithelium of specimen MZUP PL 2079 in sagittal section. Internal morphology Creeping sole 78% of body width. Glandular margin absent. Three typical geoplaninid cutaneous muscle layers present, being as thick as 8% of body height at pre-pharyngeal region. Longitudinal muscle fibres of the innermost layer (35–50 µm thick) arranged into bundles with 17–30 fibres each. Three parenchymal muscle layers present: a dorsal layer of decussate diagonal fibres, a loose supraintestinal layer of transverse and longitudinal fibres, and a loose subintestinal layer with traverse and longitudinal fibres (Fig. 12B, C). Ventral nerve plate present. Mouth situated at a distance from root of the pharynx equivalent to 66% of pharyngeal pocket length. Pharynx cylindrical (Fig. 12D). Oesophagus-to-pharynx ratio, 26%. Outer pharyngeal musculature consisting of a subepithelial layer (5 µm) of longitudinal muscle, followed by a layer (70 µm) of intermingled circular and longitudinal muscle fibres. Inner pharynx musculature consisting of a thin subepithelial layer of longitudinal muscle, followed by a layer (90 µm) of circular fibres and an innermost layer (15 µm) of longitudinal fibres. Testes dorsally located between supraintestinal parenchymal muscle layer and intestine; anteriormost testes (at a distance from anterior end equal to 22% of body length) behind ovarian region, posteriormost near root of pharynx. Penis bulb elongated, strongly developed, consisting of tightly packed muscle fibres variously orientated in its anterior section, while posteriorly the fibres are orientated diagonally, extending from 1 mm (holotype) anterior to penis papilla to the level of the gonopore, even more posteriorly in the dorsal body region. Diagonal fibres on the right side of the bulb run obliquely downwards to embrace ventrally the anterior portion of the female atrium; further course of these fibres could not be determined owing to tissue damage. Sperm ducts run dorsolaterally to the ovovitelline ducts. Laterally to penis bulb, sperm ducts bend anteriorly and medially, and then ventrolaterally penetrate the bulb. The ducts subsequently recurve and join to form an unpaired, sinuous sperm duct directed posteriorly (Fig. 13A, B). The unpaired portion continues as an ejaculatory duct inside the penis papilla, which traverses centrally to open at its tip. Sperm present in sperm duct, both along its course outside and in part of anteriorly directed portion inside penis bulb. Epithelium of sperm ducts and unpaired sperm duct pierced by glands producing minute weakly cyanophil granules; surrounded by a 7-µm-thick layer of circular muscles. Ejaculatory duct lined by 15 µm, ciliated epithelium pierced by abundant glands producing erythrophil granules; surrounded by a layer of circular muscle fibres constituting a muscular cylinder with a diameter equivalent to 0.5 times the diameter of this duct. Figure 13. View largeDownload slide Geoplana paranapiacaba Almeida & Carbayo sp. nov. A, diagrammatic representation of the copulatory apparatus of specimen MZUSP PL 2079 from sagittal sections. B, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2079. C, photomicrograph of a sagittal section of the penis papilla tip of specimen MZUSP PL 2079. D, photomicrograph of posterior section of female atrium of specimen MZUSP PL 2079. Figure 13. View largeDownload slide Geoplana paranapiacaba Almeida & Carbayo sp. nov. A, diagrammatic representation of the copulatory apparatus of specimen MZUSP PL 2079 from sagittal sections. B, photomicrograph of a sagittal section of the copulatory apparatus of specimen MZUSP PL 2079. C, photomicrograph of a sagittal section of the penis papilla tip of specimen MZUSP PL 2