taxonID	type	description	language	source
03F7B26EFFD2FFB5B1FB1E4FAEB9FAED.taxon	description	(Figures 2 – 7) Geoplanidae 4: Leal-Zanchet & Carbayo, 2000 Geoplanidae 2: Carbayo et al., 2002 Geoplana sp. 4: Leal-Zanchet & Baptista, 2009 Geoplana sp. 4: Leal-Zanchet et al., 2011	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD2FFB5B1FB1E4FAEB9FAED.taxon	etymology	Etymology The specific name refers to the dorsal pigmentation, which is yellow-ochre. Type-material Holotype. MZUSP PL. 1564: leg. P. K. Boll, 1 August 2013, São Francisco de Paula (National Forest of São Francisco de Paula), RS, Brazil – anterior tip: transverse sections on 21 slides; anterior region at the level of the ovaries: sagittal sections on 52 slides; prepharyngeal region: transverse sections on nine slides; pharynx: sagittal sections on 34 slides; copulatory apparatus: sagittal sections on 26 slides. Paratypes. São Francisco de Paula, RS, Brazil – MZU PL. 00187: leg. R. Murowaniecki, 4 May 1998, National Forest of São Francisco de Paula – copulatory apparatus: sagittal sections on 26 slides; MZU PL. 00188: leg. F. Carbayo, 8 June 1998, National Forest of São Francisco de Paula – anterior tip: transverse sections on 22 slides; region at the level of the ovaries: sagittal sections on 67 slides; pre-pharyngeal region: transverse sections on eight slides; pharynx: sagittal sections on 24 slides; copulatory apparatus: sagittal sections on 22 slides; MZU PL. 00189: leg. F. Carbayo, 25 September 1998, National Forest of São Francisco de Paula – copulatory apparatus: horizontal sections on 22 slides; MZU PL. 00190: leg. C. Palácios 17 October 2004, Research and Conservation Center Pró- Mata – pre-pharyngeal region: transverse sections on seven slides; pharynx: sagittal sections on 15 slides; copulatory apparatus: sagittal sections on 12 slides.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD2FFB5B1FB1E4FAEB9FAED.taxon	diagnosis	Diagnosis Specimens of Cratera with yellow-ochre dorsal colour mainly with dispersed greyish or greyish-brown pigmentation constituting two broad dorsal bands; eyes dorsal; glandular margin with at least four types of glands; mc: h, 11 – 12 %; pharynx cylindrical; prostatic vesicle extrabulbar with proximal portion ample and laterally expanded and distal portion tubular; ovovitelline ducts emerging dorsally from the median third of ovaries and ascending anterior to the gonopore. Molecular diagnosis. This species includes all populations that cluster with specimens KT 250622 to KT 250624 (see Supplementary material, Table S 1), with significant support in phylogenetic analyses. Type-locality São Francisco de Paula (National Forest of São Francisco de Paula), state of Rio Grande do Sul (RS), Brazil.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD2FFB5B1FB1E4FAEB9FAED.taxon	distribution	Distribution Rio Grande do Sul (São Francisco de Paula), Brazil.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD2FFB5B1FB1E4FAEB9FAED.taxon	description	Description External features. Body broad with parallel margins and dorsal surface convex; anterior tip rounded and posterior tip pointed (Figure 2 A). When creeping, maximum length reaches 85 mm. After fixation, maximum length was 52 mm (Table 1). Mouth and gonopore located at the posterior third of the body (Table 1). In living animals, dorsal surface with dispersed greyish or greyish-brown pigmentation constituting two broad dorsal bands; the dorsal yellow-ochre ground colour is mainly visible between the dorsal bands and on the body margins. The ground colour is darker on the body margins than on the rest of the dorsal surface. The dorsal pigmentation concentrates at the anterior extremity (Figure 2 A). Under the stereomicroscope, a greyish pigmentation forms two densely pigmented dorsal bands, as well as contours the anterior tip (Figures 2 B, 3 A). Posterior to the anterior tip, this pigmentation becomes more irregular and loosely arranged in flecks over the dorsum, making the yellow-ochre ground colour visible (Figures 2 B, 3 B). These flecks form the dorsal bands as well as paramarginal stripes. The dorsal bands become inconspicuous towards the posterior tip, where the pigmentation is more irregularly distributed (Figure 3 B). Ventral surface pale yellow. After fixation, the dorsal pigmentation becomes greyish and mainly greyish-brown next to the margins; dorsal ground colour becomes pale yellow, as well as the ventral surface of the body. Eyes monolobated. They are initially uniserial, with pigment cups of about 15 μm in diameter, surrounding anterior tip (Figure 3 C). After the second millimetre, the monolobated eyes become larger (with pigment cups of about 30 µm in diameter) and spread onto the dorsal surface, occupying the maximum width of about 40 % of the body width on each side of the body. Eyes remain dorsal, but become less numerous towards posterior tip (Figure 3 D). Inconspicuous clear halos may occur around dorsal eyes (Figure 3 B). Sensory organs, epidermis and body musculature. Sensory pits (Figure 4 A, B), as simple invaginations, contour anterior tip and occur ventromarginally in an irregular, single row in the anterior eighth of the body. Initially they occur at intervals of approximately 18 µm, becoming gradually sparser. Their depth is about 25 – 50 µm. Creeping sole occupying the whole body width. Two types of glands discharge through dorsal epidermis and body margins of the pre-pharyngeal region: numerous rhabditogen cells with xanthophil secretion (rhammites) and cells with amorphous, cyanophil secretion (Figure 4 E). Creeping sole receives the secretion from amorphous, cyanophil glands, as well as few rhabditogen cells with small, xanthophil rhabdites and cells with coarse granular, xanthophil secretion (Figure 4 F). The glandular margin (Figure 4 C, D), which is visible after the second millimetre of the body, receives openings of at least four types of glands. Two of them are more numerous: glands with erythrophil, coarse granules, and glands with xanthophil, coarse granules. Other two types are scarcer: cyanophil glands with amorphous secretion and erythrophil glands with fine granules. On anterior tip, glands with cyanophil, amorphous secretion and rhabditogen glands with xanthophil secretion open through the whole surface of the body and glands with coarse granular, xanthophil secretion have numerous openings through the ventral epidermis (Figure 4 A). Cutaneous musculature with the usual three layers (circular, oblique and longitudinal layers), longitudinal layer with thick bundles (Figure 4 C – F, Table 2). Cutaneous musculature thinner in the pre-pharyngeal region than in the anterior region of the body, gradually diminishing its thickness towards anterior tip (Figure 4 A). Musculature becoming progressively lower towards body margins. Ventral musculature slightly higher than dorsal at the sagittal plane. Ratio of mc: h 11 – 12 % (Table 2). Mesenchymal musculature (Figure 4 C, E, F) well developed, mainly composed of three layers: (1) dorsal subcutaneous, located mainly close to the cutaneous musculature, with oblique fibres variously oriented (about 5 – 14 fibres thick); (2) supra-intestinal transverse (about 7 – 14 fibres thick); (3) subintestinal transverse (about 14 – 22 fibres thick). In addition, there are scattered transverse subneural fibres and ventral subcutaneous oblique fibres, as well as numerous dorso-ventral fibres. On the anterior region of the body (Figure 4 A, B), the mesenchymal musculature is less developed than in the prepharyngeal region. Pharynx. Pharynx cylindrical, about 5 – 6 % of body length, with dorsal insertion posteriorly shifted, but located at the anterior third of pharyngeal pouch. Mouth posterior to the dorsal insertion, in the median third of pharyngeal pouch (Figure 4 G). Oesophagus short; oesophagus: pharynx ratio varying from 14 % in paratype MZU PL. 00190 to 18.5 % in the holotype. It is lined by ciliated, cuboidal to columnar epithelium with some insunk nuclei. The oesophagus is coated with a thick subepithelial muscle layer with circular fibres, followed by a thin muscle layer with longitudinal fibres (about 60 – 100 µm thick). Pharynx and pharyngeal lumen lined by ciliated, cuboidal epithelium with insunk nuclei. Pharyngeal glands constituted by three secretory cell types: cells with fine granular erythrophil secretion; cells with fine granular xanthophil secretion and cells with amorphous cyanophil secretion. Cell bodies of pharyngeal glands located in the mesenchyme. Outer pharyngeal musculature (about 10 – 24 µm thick) comprised of subepithelial layer of longitudinal muscles, followed by a circular layer and more internally by longitudinal fibres. All layers become thinner towards pharyngeal tip and the longitudinal internal fibres become mixed with the circular fibres. Inner pharyngeal musculature (about 100 – 150 µm thick) comprises a thick subepithelial layer with circular fibres, mixed internally and externally with longitudinal fibres. Inner musculature gradually becomes thinner towards pharyngeal tip. Reproductive organs. Testes in one irregular row on either side of the body, located beneath the dorsal transverse mesenchymal muscles (Figure 4 C, E). They begin slightly posterior to the ovaries, in the anterior third of the body, and extend to near the root of the pharynx (Table 1). Sperm ducts medial to ovovitelline ducts, above fibres of the subintestinal transverse mesenchymal musculature, in pre-pharyngeal region. They form spermiducal vesicles posterior to pharynx. Distally, spermiducal vesicles enter laterally into the proximal expanded portion of the prostatic vesicle (Figures 5 A, B, 7 A). Extrabulbar prostatic vesicle, unpaired, located near the common muscle coat, with ample proximal portion and tubular distal portion. The proximal portion is laterally expanded and T-shaped (Figures 5 B, 7 A); it is located closer to the ventral epidermis than to the dorsal epidermis. The prostatic vesicle penetrates the common muscle coat, becoming sinuous, and continues inside the penis papilla as an ejaculatory duct (Figure 5 A). Ejaculatory duct slightly sinuous, opening through an expansion at the tip of the penis papilla (Figure 5 B, 7 B). Male atrium without folds, occupied by the conical and symmetric penis papilla with ventral insertion posteriorly displaced (Figures 5 A, B, 6 A, B, Table 1). Lining epithelium of sperm ducts cuboidal and ciliated; thin muscularis (about 2 – 4 µm) constituted of interwoven circular and longitudinal fibres. Prostatic vesicle lined with ciliated, tall columnar or pseudostratified epithelium. Muscularis of prostatic vesicle (about 10 – 20 µm thick) comprises circular fibres mixed with longitudinal and oblique fibres. Ejaculatory duct lined with ciliated, columnar epithelium, showing a pseudostratified appearance at the expanded portion (Figure 7 B). Muscle coat of ejaculatory duct thin (about 3 – 5 µm), mainly constituted of circular fibres. Both prostatic vesicle and ejaculatory duct receive openings from glands producing a fine granular, mixed secretion (Figure 7 A) as well as glands with amorphous cyanophil secretion. Granules with a mixed secretion have a cyanophil external part and an erythrophil internal core. Both glands with extrabulbar cell bodies. Penis papilla lined with non-ciliated, pseudostratified epithelium. Penis glands produce numerous fine granular, xanthophil and erythrophil secretions as well as less numerous cyanophil secretions of two types (amorphous and fine granular secretion). All penis glands with cell bodies external to common muscle coat; their long necks run longitudinally through the papilla (Figures 6 A, B, 7 B), with numerous openings through its lining epithelium. Muscularis of the penis papilla (about 6 – 10 µm thick) composed of a subepithelial circular layer, followed by a longitudinal layer. Epithelial lining of male atrium columnar to pseudostratified (about 20 – 150 µm thick), non-ciliated. Four types of glands empty through this epithelium: abundant cells with fine granular, cyanophil secretion and cells with cyanophil amorphous secretion; and few glands with fine granular, mixed secretion (cyanophil peripheral portion and erythrophil core) as well as scarce xanthophil glands. These glands have their cell bodies located in the mesenchyme, mainly anterior and laterally to the copulatory apparatus, or between the fibres of the common muscle coat. Numerous necks of both types of cyanophil glands concentrate their openings at the dorso-lateral wall of the male atrium (Figure 6 A, B). Their numerous gland necks are located between fibres of the stroma in the dorso-lateral walls of the male atrium. Muscularis of male atrium (about 4 – 8 μm thick) comprised of a subepithelial layer with circular fibres, followed by a longitudinal layer. Vitellaria (Figure 4 C, F), situated between intestinal branches, opening into the ovovitelline ducts. Ovaries oval-elongate (Figure 4 H), measuring about 300 – 520 µm in diameter in the holotype and paratype MZU PL. 00188, respectively. They are located dorsal to the ventral nerve plate, in the anterior third of the body (Table 1). Ovovitelline ducts emerge dorsally from the median third of the ovaries (Figure 4 H) and run posteriorly immediately above the nerve plate. Lateral to the female atrium, ovovitelline ducts ascend postero-medially, to unite dorsal to the end of the median third or dorsal to the posterior third of the female atrium, thus forming the common glandular ovovitelline duct (Figures 5 A, B, 6 A). The female genital duct is dorso-anteriorly curved (Figures 5 A, B, 6 A). Female atrium, shorter than male atrium, funnel-shaped without folds (Figures 5 A, B, 6 A, B, Table 1). Ovovitelline ducts and common ovovitelline duct lined with ciliated, cuboidal to columnar epithelium and covered with intermingled circular and longitudinal muscle fibres (about 3 – 5 μm). Numerous shell glands with erythrophil secretion empty into common glandular ovovitelline duct as well as into the distal half of the ascending portion of the ovovitelline ducts (Figures 5 A, B, 6 A, B). Female genital duct and atrium lined by tall columnar to pseudostratified epithelium (Figure 6 A) containing some lacunae. Female duct and atrium receive abundant cyanophil secretion of two types (amorphous and fine granular secretion), as well as erythrophil glands and few xanthophil glands. Their cell bodies are located between fibres of the atrial stroma or external to the common muscle coat. Muscularis of vagina and female atrium (about 15 µm thick) composed mainly of circular fibres interposed with some longitudinal fibres. Gonoduct vertical at the sagittal plane. Male and female atria with ample communication, without separating folds (Figures 5 A, B, 6 A, B). Gonoduct lined with ciliated columnar epithelium, receiving the openings of numerous rhabditogen glands and cyanophil glands with amorphous secretion. Muscularis of gonoduct comprised of a subepithelial layer of circular fibres, followed by a longitudinal layer. Common muscle coat with circular, longitudinal and oblique fibres; it is thin along both male and female atria. A stroma with sparse intermingled muscle fibres separates the atrial muscularis and common muscle coat. Variability Paratype MZU PL. 000190 is at an initial stage of maturity, showing poorly developed testes and vitellaria, few shell glands and short female atrium. In paratype MZU PL. 00187, the penis papilla is protruded towards the gonoduct and there is abundant cyanophil secretion in the male atrium and in the gonoduct; the epithelial lining of the female atrium is taller laterally than medially. The proximal portion of the prostatic vesicle contains sperm in paratype MZU PL. 00189. Comparative discussion In accordance with the phylogenetic analyses, the external and internal characters of the new species herein described as Cratera ochra sp. nov. strongly support its inclusion into the genus Cratera Carbayo et al. The morphological characters concern the following features: monolobated eyes, ejaculatory duct forming a distal cavity in the penis papilla and funnel-shaped female atrium. Cratera ochra sp. nov., with dorsal eyes, can be differentiated from Cratera pseudovaginuloides (Riester, 1938), Cratera yara (E. M. Froehlich, 1955) and Cratera cuarassu Carbayo & Almeida, 2015, which have exclusively marginal eyes (E. M. Froehlich 1955; Froehlich 1956; Carbayo and Almeida 2015). It is also easily distinguished from Cratera steffeni Rossi et al., 2014, in which the eyes are restricted to either side of the body (Rossi et al. 2014). These four species and also Cratera crioula (E. M. Froehlich, 1955), Cratera joia (Froehlich, 1956) and Cratera anamariae Carbayo & Almeida, 2015 have different colour patterns in comparison to the new species herein described (Riester 1938; E. M. Froehlich 1955; Froehlich 1956; Rossi et al. 2014; Carbayo and Almeida 2015). Cratera ochra sp. nov. has colour and eye patterns similar to those of Cratera tamoia (E. M. Froehlich, 1955), but the new species can be mainly distinguished from the latter by its cylindrical pharynx, whereas the pharynx of C. tamoia is bell-shaped (E. M. Froehlich 1955). With respect to the morphology of the copulatory apparatus, species of the genus Cratera have an extrabulbar prostatic vesicle, a penis papilla with a distal cavity and a funnel-shaped female atrium, among other characteristics. The prostatic vesicle has two portions, a proximal expanded section and a distal tubular and sinuous portion. In C. ochra sp. nov., similarly to C. steffeni, the proximal portion is characteristically T-shaped with the sperm ducts opening through its lateral walls. This form is particularly evident in horizontal sections of the copulatory apparatus. Other species of the genus, such as C. crioula, C. joia, C. pseudovaginuloides and C. tamoia, which were described as possessing a prostatic vesicle with proximal diverticula (Riester 1938; E. M. Froehlich 1955; Froehlich 1956), may have a vesicle with a similar form. In summary, C. ochra sp. nov. is mainly differentiated from its congeners by the combination of the following morphological characters: dorsal body surface with dispersed pigmentation forming two broad bands, eyes spreading over the dorsal surface, cylindrical pharynx. Measures of intraspecific and interspecific variation of the COI gene, as well as the ABGD algorithm applied to the COI data set, supported C. ochra sp. nov. as a species different from its congeners. In addition, both phylogenetic methods (maximum likelihood and Bayesian inference) revealed similar topologies, placing C. ochra sp. nov. as the sister group of the clade formed by C. tamoia and C. crioula (Figure 1).	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD8FFA0B1A219C8AEC6FD7A.taxon	description	(Figures 8 – 11) Geoplana sp. 5: Leal-Zanchet & Carbayo, 2000 Geoplana sp. 1: Carbayo et al., 2002 Geoplana sp. 2: Baptista et al., 2006 Geoplana sp. 2: Fick et al., 2006 Geoplana sp. 3: Leal-Zanchet & Baptista, 2009 Geoplana sp. 3: Leal-Zanchet et al., 2011	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD8FFA0B1A219C8AEC6FD7A.taxon	etymology	Etymology The specific name refers to the dorsal pigmentation, which forms numerous flecks and dots.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD8FFA0B1A219C8AEC6FD7A.taxon	materials_examined	Type material Holotype. MZUSP PL. 1565: leg. P. K. Boll, 10 April 2014: São Francisco de Paula (National Forest of São Francisco de Paula), RS, Brazil – anterior tip: transverse sections on 21 slides; anterior region at the level of the ovaries: sagittal sections on 72 slides; prepharyngeal region: transverse sections on 20 slides; pharynx: sagittal sections on 36 slides; copulatory apparatus: sagittal sections on 40 slides. Paratypes. MZU PL. 00193: leg. R. Murowaniecki, 25 November 1997: São Francisco de Paula (National Forest of São Francisco de Paula), RS, Brazil – anterior tip: transverse sections on six slides; anterior region at the level of the ovaries: sagittal sections on 12 slides; pre-pharyngeal region: transverse sections on 11 slides; pharynx: sagittal sections on 42 slides; copulatory apparatus: sagittal sections on 36 slides. MZU PL. 00194: leg. F. Carbayo, 11 December 1997: São Francisco de Paula (National Forest of São Francisco de Paula), RS, Brazil – anterior tip: transverse sections on 18 slides; anterior region at the level of the ovaries: sagittal sections on 42 slides; pre-pharyngeal region: transverse sections on 15 slides; pharynx: sagittal sections on 29 slides; copulatory apparatus: sagittal sections on 34 slides. MZU PL. 00195: leg. I. A. Fick, 25 November 1998: São Francisco de Paula (National Forest of São Francisco de Paula), RS, Brazil – pre-pharyngeal region: transverse sections on 12 slides; pharynx: sagittal sections on 36 slides; copulatory apparatus: horizontal sections on 34 slides. MZU PL. 00196: leg. I. A. Fick, 23 June 2000: Praia Grande (National Park of Aparados da Serra), SC, Brazil – pre-pharyngeal region: transverse sections on 12 slides; pharynx: sagittal sections on 36 lâminas; copulatory apparatus: sagittal sections on 34 slides. MZU PL. 00197: leg. S. V. Amaral, 21 March 2010: São Francisco de Paula (National Forest of São Francisco de Paula), RS, Brazil – anterior region preserved on clove oil; copulatory apparatus: sagittal sections on 27 slides. Type-locality National Forest of São Francisco de Paula, São Francisco de Paula, state of Rio Grande do Sul (RS), Brazil.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD8FFA0B1A219C8AEC6FD7A.taxon	distribution	Distribution States of Rio Grande do Sul (São Francisco de Paula) and Santa Catarina (Praia Grande), Brazil.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD8FFA0B1A219C8AEC6FD7A.taxon	diagnosis	Diagnosis Specimens of Obama with brown dorsum, sometimes greyish-brown, with a marbled appearance; live specimens with pale-yellow ventral surface in the anterior third and orange in the median and posterior thirds of the body; eyes spreading over the lateral parts of the dorsal surface; glandular margin with three types of glands; mc: h, 10 – 12 %; pharynx cylindrical; prostatic vesicle with ovoid and unbranched proximal portion; penis papilla symmetrical and truncated with an almost straight ejaculatory duct; ovovitelline ducts emerging dorsally from the median third of ovaries and ascending anterior to the gonopore. Molecular diagnosis. This species includes all populations that cluster with specimens KT 250625 to KT 250629 (see Supplementary material, Table S 1), with significant support in phylogenetic analyses.	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
03F7B26EFFD8FFA0B1A219C8AEC6FD7A.taxon	description	Description External features. Body lanceolate, elongate and flat; anterior end more narrowed than the posterior end (Figure 8 A). When crawling, maximum length reaches 72 mm (Table 3). Mouth and gonopore located at the posterior third of the body, with the exception of the mouth in paratype MZU PL. 00197, which is located at the median third of the body (Table 3). Live specimens with brown dorsum, sometimes greyish-brown, with a marbled appearance (Figure 8 A); ventral surface pale-yellow in the anterior third and orange in the median and posterior thirds of the body. Under moderate magnification, the anterior tip shows two median dark stripes, whereas the rest of the dorsum shows irregular greyish flecks laterally, over a light brown ground colour, and numerous dark-grey spots medially (Figure 8 B). Under the stereomicroscope, dorsal ground-colour greyish, covered by fine brownish pigmentation with dark grey spots in a wide median zone of the body and flecks on the body margins. Flecks form two median stripes on the anterior and posterior tips; these stripes are more parallel to each other anteriorly (Figure 8 B). In preserved specimens dorsal and ventral colour fade, the ventral surface becoming pale yellow. Eyes, initially uniserial, surround anterior tip and become pluriserial immediately after this tip. After the fifth millimetre of the body, eyes spread over the lateral parts of the dorsum, forming a maximum of five longitudinal rows on either side of the dorsal surface (Figure 8 C). They are monolobated, with pigment cups of about 30 – 60 μm in diameter, on the anterior third of the body (Figure 8 D). After that, they are trilobated (with pigment cups of about 20 – 40 μm in diameter), occupying the lateral parts of the dorsal surface (Figure 8 E), but become less numerous towards posterior tip. Clear halos around eyes are absent. Sensory organs, epidermis and body musculature. Sensory pits (Figure 9 A, B), as simple invaginations, contour anterior tip and occur ventromarginally in an irregular, single row in the anterior seventh of the body. Initially they occur at intervals of approximately 12 µm, becoming gradually sparser. Their depth is about 25 – 40 µm. Creeping sole occupying the whole body width. Three types of glands discharge through dorsal epidermis and body margins of the pre-pharyngeal region: numerous rhabditogen cells with xanthophil secretion (rhammites) and erythrophil cells with fine granular secretion, as well as few glands with amorphous, cyanophil secretion (Figure 9 E). Creeping sole receives the secretion from numerous coarse granular, cyanophil glands (Figure 9 F), as well as few rhabditogen glands with small, xanthophil rhabdites and cells with fine granular, erythrophil secretion. After the sixth millimetre of the body, glandular margin (Figure 9 C, D) receiving openings of at least three types of glands: abundant glands with xanthophil, coarse granules, as well as scarcer erythrophil glands with fine granules and cyanophil glands with amorphous secretion. Some rhabditogen cells with small, xanthophil rhabdites open between cells of the glandular margin. On anterior tip (Figure 9 A, B), glands with cyanophil secretion and scarce glands with fine granular, erythrophil secretion open through the whole surface of the body. In addition, numerous glands with coarse granular, xanthophil secretion open through the ventral epidermis and rhabditogen glands open through the dorsal epidermis and body margins. Cutaneous musculature with the usual three layers (circular, oblique and longitudinal layers), longitudinal layer with thick bundles (Figure 9 C – F, Table 4). Cutaneous musculature thinner in the pre-pharyngeal region than in the anterior region of the body, but gradually diminishing towards anterior tip. Musculature higher medially, becoming progressively lower towards body margins; ventral musculature higher than dorsal at the sagittal plane in the pre-pharyngeal region mc: h 11 – 12 % (Table 4). Mesenchymal musculature (Figure 9 C, E, F) mainly composed of three layers: (1) dorsal subcutaneous, located mainly close to the cutaneous musculature, with oblique fibres variously oriented (about three to four fibres thick); (2) supra-intestinal transverse (about 11 – 16 fibres thick); (3) subintestinal transverse (about seven to eight fibres thick). In addition, there are scattered ventral subcutaneous oblique fibres and transverse subneural fibres, as well as dorso-ventral fibres. On the anterior region of the body (Figure 9 A, B), the mesenchymal musculature is less developed than in the pre-pharyngeal region. Pharynx. Pharynx cylindrical with dorsal insertion shifted posteriorly, about 6 – 7 % of the body length, occupying the first two-thirds of the pharyngeal pouch; mouth in the posterior third of pharyngeal pouch (Figure 9 G). Oesophagus short; with folded wall. Oesophagus: pharynx ratio varying from 5 % to 24 % (10 % in the holotype). It is lined by ciliated, cuboidal epithelium with insunk nuclei and coated with a thick muscle layer with circular fibres and various interposed longitudinal fibres (about 40 – 70 µm thick). Pharynx and pharyngeal lumen lined by ciliated, cuboidal epithelium with insunk nuclei. Pharyngeal glands constituted by three secretory cell types: abundant glands with fine granular erythrophil secretion; glands with fine granular xanthophil secretion; and glands with amorphous cyanophil secretion. Cell bodies of pharyngeal glands located in the mesenchyme, mainly anterior and posterior to the pharynx. Outer pharyngeal musculature (about 20 – 30 µm thick) comprised of thin subepithelial layer of longitudinal muscles, followed by a thicker layer of circular fibres. Inner pharyngeal musculature (about 50 – 110 µm thick) comprises a thick subepithelial layer of circular fibres, interposed with various longitudinal fibres. Outer and inner musculatures gradually become thinner towards pharyngeal tip. Reproductive organs. Testes in at least three irregular rows beneath the dorsal transverse mesenchymal muscles (Figure 9 C, E). They begin in the anterior third of the body and extend to near the root of the pharynx (Table 3). Sperm ducts dorsal to ovovitelline ducts, in pre-pharyngeal region subdivided into three or four ductules. They form spermiducal vesicles posterior to the pharynx. Distally, spermiducal vesicles penetrate into the lateral wall of the proximal portion of the prostatic vesicle (Figures 10 A, B, 11 B). Prostatic vesicle extrabulbar, unpaired, and consisting of two portions: a globose proximal portion and a tubular, sinuous, distal portion (Figure 10 A, B). The tubular portion penetrates the common muscle coat and continues inside the penis papilla as an ejaculatory duct. Ejaculatory duct almost straight, opening at the tip of a truncated penis papilla (Figures 10 A, B, 11 A, B). The tip of the penis papilla may form a globose invagination with irregular contour (Figures 10 A, 11 A). Male atrium almost without folds, occupied by the conical and symmetrical penis papilla (Table 3, Figures 10 A, B, 11 A, B). The latter may occupy the distal part of the female atrium, as in paratypes MZU PL. 00196 and MZU PL. 00197. Lining epithelium of sperm ducts cuboidal and ciliated; thin muscularis (about 6 – 10 µm thick) mainly constituted of circular fibres. Prostatic vesicle lined with ciliated, columnar or pseudostratified epithelium with irregular height. It receives abundant amorphous cyanophil secretions and scarcer fine granular erythrophil secretions, both from secretory cells with bodies lying in the mesenchyme, mainly around the vesicle. Muscularis of prostatic vesicle (about 50 – 120 µm thick) comprises interwoven longitudinal, oblique and circular fibres (Figure 11 B). Ejaculatory duct lined with ciliated, columnar epithelium, receiving few openings from secretory cells with weakly cyanophil, amorphous secretion. Muscle coat of ejaculatory duct thin (about 10 – 20 µm thick), constituted of interwoven circular and longitudinal fibres. Penis papilla covered with non-ciliated, columnar epithelium, becoming lower towards the tip of the papilla (Figure 11 A, B). Penis glands of four types: cells with numerous fine granular erythrophil secretion, cells with mixed (cyanophil peripheral portion and erythrophil core), fine granular secretion and scarcer glands with cyanophil secretion and glands with xanthophil secretion. They run longitudinally in the papilla, with numerous openings through its covering epithelium, the erythrophil glands concentrating their openings at the tip and ventral surface of the penis papilla (Figure 11 A). Penis glands with cell bodies mainly external to common muscle coat and among fibres of this coat. Muscularis of the penis papilla (about 30 – 60 µm thick) composed of a thick, subepithelial layer of circular fibres interposed with some longitudinal fibres. Epithelial lining of male atrium columnar (about 10 – 25 µm thick), non-ciliated. Three types of secretory cells empty through this epithelium: glands with fine granular, erythrophil secretion, glands with amorphous cyanophil secretion and glands with fine granular mixed secretion (cyanophil peripheral portion and erythrophil core). Numerous necks of the latter concentrate their openings at the dorsal wall of the distal portion of the male atrium (Figure 11 A). Muscularis of male atrium (about 15 – 45 µm thick) comprised of circular fibres and some interposed longitudinal fibres. Vitellaria (Figure 9 C, E, F), situated between intestinal branches, open into the ovovitelline ducts. Ovaries ovoid or oval-elongate (Figure 9 H), measuring about 400 µm in diameter in the holotype and 200 – 300 µm in diameter in paratypes MZU PL. 00194 and MZU PL. 00193, respectively. They are dorsal to the ventral nerve plate, at the same transverse level as the anteriormost testes or slightly anterior to them. Ovovitelline ducts emerge dorsally (Figure 9 H) from the median third of the ovaries and run posteriorly immediately above the nerve plate. Lateral to the female atrium, ovovitelline ducts ascend postero-medially, to unite dorsal to the median or posterior third of female atrium, so forming a short common glandular ovovitelline duct. Ental portion of female atrium with an antero-dorsally directed female canal (Figures 10 A, B, 11 A). Female atrium ovoid with folded walls (Figures 10 A, B, 11 A, B). Female atrium length about half that of male atrium in the holotype, but almost the same as male atrium length in most paratypes (Table 3). Ovovitelline ducts and common oviduct lined with ciliated, cuboidal to columnar epithelium. They are covered with intermingled circular and longitudinal muscle fibres (about 20 – 30 μm thick). Abundant shell glands with erythrophil secretion empty into the common glandular ovovitelline duct as well as in the distal half of the ascending portion of the ovovitelline ducts (Figures 10 A, B, 11 A, B). Female atrium and canal lined by tall columnar epithelium, exhibiting a multilayered aspect in some places (Figure 11 A) and with a xanthophil apical layer. Female atrium and canal receive abundant fine granular erythrophil secretion and cyanophil amorphous secretion. Cell bodies of both glands are located external to the common muscle coat or among its fibres. Muscularis of female atrium and canal (about 20 – 40 µm thick) composed of a subepithelial layer of interwoven circular and longitudinal fibres. Gonoduct vertical at the sagittal plane. Male and female atria with ample communication, without separating folds (Figures 10 A, B, 11 A, B). Gonoduct lined with ciliated columnar epithelium, receiving the openings of abundant glands with fine granular erythrophil secretion and glands containing amorphous cyanophil secretion, as well as rhabditogen glands with small rhabdites. Muscularis of gonoduct consisting of a subepithelial layer of circular fibres, followed by a layer of longitudinal fibres. Common muscle coat poorly developed, with circular, longitudinal and oblique fibres, thicker around male atrium than around female atrium. A stroma with sparse intermingled muscle fibres separates the atrial muscularis from the common muscle coat. Comparative discussion In general, external and internal characteristics of O. maculipunctata sp. nov. match the diagnostic characteristics of the genus Obama Carbayo et al., 2013, so corroborating the molecular phylogenetic analyses. The new species of Obama herein described shows an internal morphology similar to four other species of this genus, namely Obama fryi (von Graff, 1899), Obama polyophthalma (von Graff 1899), Obama carinata (Riester, 1938) and Obama eudoximariae (Ogren & Kawakatsu, 1990). These species share an extrabulbar prostatic vesicle with a globose proximal portion, a symmetrical penis papilla with dorsal and ventral insertions at the same transverse level, and an ovoid female atrium (von Graff 1899; Schirch 1929; Riester 1938; Marcus 1951; Froehlich 1956; Ogren and Kawakatsu 1990). Considering external features, by having a marbled colour pattern, the new species of Obama is similar to O. polyophthalma. It differs from O. fryi, O. carinata and O. eudoximariae, because they have homogeneous or striped patterns (von Graff 1899; Schirch 1929; Riester 1938; Marcus 1951; Froehlich 1956). Regarding the anatomy, O. maculipunctata, which has a cylindrical pharynx, can be differentiated from O. carinata and O. eudoximariae which have a collar-type pharynx (Schirch 1929; Riester 1938; Marcus 1951; Ogren and Kawakatsu 1990). In addition, O. maculipunctata differs from all four species by a combination of the following characteristics of the copulatory apparatus: unbranched proximal portion of the prostatic vesicle, truncated penis papilla, and almost straight ejaculatory duct traversing the penis papilla. As a number of species of Geoplana incertae sedis have general characteristics similar to species of Obama, we have extended our comparative analysis of O. maculipunctata to this group of species. Among the 47 species indicated by Carbayo et al. (2013) as Geoplana incertae sedis, only one species, Geoplana fuhrmanni Hyman, 1962, from Panama, shows characteristics of the copulatory apparatus similar to those of O. maculipunctata. However, having a marbled dorsum, the new species can be distinguished from G. fuhrmanni, with an almost homogeneous, spotted coloured dorsal surface, as well as by the morphology of the female atrium, which is short and funnelshaped, in contrast to the large and ovoid female atrium of O. maculipunctata. Comparison of the new species with its congeners, using measures of intraspecific and interspecific variation of the COI gene, as well as the ABGD tool applied to the COI data set, also supported O. maculipunctata sp. nov. as a separate species. Maximum likelihood and Bayesian inference phylogenies were congruent and indicated O. maculipunctata sp. nov. as the sister group of O. carinata (Figure 1). Notes on ecology and distribution Cratera ochra sp. nov. has been recorded mainly inside and on the borders of sites with the native vegetation (mixed ombrophilous forest) of its type-locality (Leal-Zanchet and Carbayo 2000; Carbayo et al. 2002; Leal-Zanchet et al. 2011), with a low to moderate abundance. It was also recorded in an area of dense ombrophilous forest, but not in areas of mixed ombrophilous forest, of the Research and Conservation Center Pró-Mata (Leal- Zanchet et al. 2011). Obama maculipunctata sp. nov. is abundant in areas of mixed ombrophilous forest of the Araucaria Plateau, as well as in plantations of the native Araucaria angustifolia and Pinus spp. of its type-locality (Leal-Zanchet and Carbayo 2000; Carbayo et al. 2002). It was also recorded in areas of mixed and dense ombrophilous forest of the Aparados da Serra National Park (Baptista et al. 2006; Fick et al. 2006), but it was not recorded in the Research and Conservation Center Pró-Mata (Leal-Zanchet et al. 2011). General discussion In general, the two new species herein described match the diagnostic characteristics of their respective genera, despite their gross similarities. Species of Cratera and Obama have a similar external morphology, with a lanceolate body, usually slightly more leafshaped in species of Obama than in species of Cratera. Species of Obama can be distinguished from Cratera, as well as from other genera split off from Geoplana, by having trilobated eyes in addition to monolobated eyes (Carbayo et al. 2013). Other anatomical features, such as the distribution pattern of the sensory pits and details of the copulatory apparatus are useful in the identification of the species. Some of these features are noteworthy especially the distribution pattern of the sensory pits and the morphology of the prostatic vesicle. In relation to the distribution pattern of the sensory pits in both genera, Carbayo et al. (2013) indicated that members of Cratera have the sensory pits arranged in a single row on either side of the body, whereas members of Obama have two to four rows of sensory pits on either side. However, we have observed that a single irregular longitudinal row of sensory pits on either side of the body is present in O. maculipunctata. As this character may be important for distinguishing monophyletic groups, it should be further investigated in other species of Obama. Another feature that should be analysed in more detail in species of Obama is the morphology of the prostatic vesicle. Various species have an anteriorly branched prostatic vesicle, but this feature may not have been properly assessed in earlier descriptions (cf. von Graff 1899; Schirch 1929; Riester 1938; Marcus 1951). In O. maculipunctata, the proximal portion of the prostatic vesicle is clearly unbranched. The genus Cratera, with eight formally described species (Carbayo et al. 2013; Rossi et al. 2014; Carbayo and Almeida 2015), had the type-species and four others molecularly analysed (Carbayo et al. 2013). In contrast, the genus Obama, originally with 34 species, was poorly investigated at the molecular level. Four species of Obama were analysed by Carbayo et al. (2013) and, more recently, two new species were proposed for the genus based on morphological and molecular studies (Álvares-Presas et al. 2015). It is noteworthy that one of the species included in the present phylogenetic analysis, O. carinata, showed a high intraspecific distance in relation to other species of the group. These results are also supported by the ABGD analysis, which split the specimens of O. carinata into two major mitochondrial DNA lineages, suggesting that they are two potential species. However, a Bayesian phylogenetics and phylogeography analysis (Yang and Rannala 2010) applied for a large data set indicated that the specimens of O. carinata constituted a single species (Álvares-Presas et al. 2015). In fact, Puillandre et al. (2012) point out that the partition output by ABGD should be considered as a first species partition hypothesis, since this result arises from the evaluation of pairwise differences of a single molecular marker. Therefore, the definition of the taxonomic status should be complemented with other evidence, such as morphological data, as done herein, in an integrative taxonomic approach (Will et al. 2005; Padial et al. 2009; Puillandre et al. 2012).	en	Rossi, Ilana, Amaral, Silvana Vargas, Ribeiro, Giovana Gamino, Cauduro, Guilherme Pinto, Fick, Israel, Valiati, Victor Hugo, Leal-Zanchet, Ana Maria (2015): Two new Geoplaninae species (Platyhelminthes: Continenticola) from Southern Brazil based on an integrative taxonomic approach. Journal of Natural History 50: 787-815, DOI: 10.1080/00222933.2015.1084057, URL: http://dx.doi.org/10.1080/00222933.2015.1084057
