Integrative approach reveals two new species of Obama (Platyhelminthes: Tricladida) from the South-Brazilian Atlantic Forest Marques, Alessandro Rossi, Ilana Valiati, Victor Hugo Leal-Zanchet, Ana Maria Zootaxa 2018 2018-08-01 4455 1 99 126 Marques & Rossi & Valiati & Leal-Zanchet, 2018 Marques & Rossi & Valiati & Leal-Zanchet 2018 [151,348,969,995] Rhabditophora Geoplanidae Obama GBIF Animalia Tricladida 2 101 Platyhelminthes species allandra sp. nov.   Etymology:The specific name is a cOmpOsite Of the Greek adjective állos(different) and Greek nOun andrós(man), referring tO the male atrium which shOws a variable aspect in the specimens studied.     Typematerial. Holotype: MZUSPPL.2139: leg.I. ROssi, 0  7 February 2015, GeneralCarneirO (  Araucaria Natural Heritage Private Reserve), state Of Paraná, Brazil—anteriOr tip: transverse sectiOns On18 slides; anteriOr regiOn at the level Ofthe Ovaries: sagittal sectiOns On26 slides; pre-pharyngeal regiOn: transverse sectiOns On15 slides; pharynx and cOpulatOry apparatus: sagittal sectiOns On76 slides.   Paratypes: MZU PL.00283: leg.I. ROssi, 0 6 February 2015, General CarneirO (  AraucariaNatural Heritage Private Reserve), state Of Paraná, Brazil—cOpulatOry apparatus: sagittal sectiOns On 26 slides. MZU PL.00284: leg.I. ROssi, 0 7 February 2015, General CarneirO (  AraucariaNatural Heritage Private Reserve), state Of Paraná, Brazil—anteriOr tip: transverse sectiOns On 19 slides; pre-pharyngeal regiOn: transverse sectiOns On 7 slides; pharynx and cOpulatOry apparatus: sagittal sectiOns On 25 slides.   Diagnosis:species Of  Obamawith dOrsal grOund cOlOur brOwnish, cOvered by numerOus irregular dark flecks, and median band yellOwish On anteriOr regiOn Of bOdy; pharynx cylindrical with dOrsal insertiOn shifted pOsteriOrly; prOstatic vesicle with twO pOrtiOns, prOximal pOrtiOn fOrked, glObOse Or OvOid, and distal pOrtiOn wide and inverted-U shaped; penis papilla either cOnical and almOst symmetrical Or irregular in shape and length; male atrium with a dOrsO-lateral invaginatiOn; male and female atria separated by dOrsal fOld.  Molecular diagnosis:this species includes all pOpulatiOns that cluster with sequences Of specimens included in this study (GenBank accessiOn numbers MH378881- MH378884) with significant suppOrt in an adequate mOlecular delimitatiOn mOdel.  Type-locality.General CarneirO, Paraná( PR), Brazil.   Distribution:knOwn Only frOm the type-lOcality, General CarneirO, PR, Brazil.   Description. External features.BOdy elOngate with parallel margins and dOrsal surface slightly cOnvex; anteriOr tip rOunded and pOsteriOr tip pOinted ( Fig. 1). Live specimens with maximum length Of 39 mmwhen creeping. After fixatiOn, maximum length 29 mm. MOuth at third fOurth Of bOdy and gOnOpOre at pOsteriOr fOurth Of bOdy ( Table 1). Live specimens with dOrsal grOund cOlOur brOwnish and median band yellOwish, which is lighter On anteriOr regiOn Of bOdy, becOming greyish On cephalic regiOn ( Fig. 1). Irregular blackish Or dark-brOwn flecks Occur Over dOrsal surface, fOrming paramedian stripes On anteriOr bOdy half and Overlaying median band On pOsteriOr half ( Figs. 1, 2). After fixatiOn, median band yellOwish, becOming incOnspicuOus tOwards pOsteriOr tip. Dark irregular flecks lOnger in anteriOr bOdy half and smaller and mOre densely distributed in pOsteriOr half, fOrming paramedian stripes in anteriOr third Of bOdy, excepting cephalic regiOn ( Figs. 2, 4–5). In live and fixed specimens, ventral surface light-brOwn with greyish pigmentatiOn cOntOuring anteriOr tip.   FIGURE 1.  Obama allandra  sp. nov., holotype: photograph in dorsal view. Anterior tip to the left.   TABLE 1.Measurements, in mm, of specimens of  Obama allandra  sp. nov.–: not measured; *: after fixation; ** length; DG: distance of gonopore from anterior end; DM: distance of mouth from anterior end; DMG: distance between mouth and gonopore; DPVP: distance between prostatic vesicle and pharyngeal pouch. The numbers given in parentheses represent the position relative to body length.    Holotype Paratype MZU PL.00283 Paratype MZU PL.00284  Maximum length in extension 39 32 25  Maximum width in extension 2 3.5 4  Length at rest 25 24 20  Width at rest 5 5 5  Length* 29 28 25  Width* 5 4.5 4  DM* 20 (69) 18.5 (66) 16 (64)  DG* 24 (83) 23 (82) 19.5 (78)  DMG* 4 4.5 3.5  DPVP* 0.8 – 0.9  Ovaries 8 (27) – –  Anteriormost testes 6 (21) – –  Posteriormost testes 18 (62) – –  Prostatic vesicle** 1.9 1.7 2.3  Penis papilla** 0.1 0.7 1  Male atrium** 1.3 1.1 1.1  Female atrium** 1.1 1.6 0.9 Eyes, initially mOnOlObate and uniserial, surrOund anteriOr tip ( Figs. 3–4). After secOnd millimetre Of bOdy, eyes becOme larger and spread OntO dOrsal surface, Occupying maximum width Of abOut One-fOurth Of bOdy width On either side Of bOdy, lateral tO median band. Eyes becOme trilObate after anteriOr fOurth Of bOdy, remaining dOrsal, but less numerOus tOwards pOsteriOr tip ( Fig. 3). Clear halOs, less nOticeable, may Occur arOund dOrsal eyes ( Fig. 5). Pigment cups abOut 20–45 µm Of diameter.  Sensory organs, epidermis and body musculature.SensOry pits ( Figs. 6, 7), as simple invaginatiOns (25–45 µm deep), cOntOur anteriOr tip and Occur ventrOmarginally in irregular, single rOw in almOst the entire anteriOr half Of bOdy (abOut 45% Of bOdy length). Glands Of three typesdischarge thrOugh whOle epidermis Of pre-pharyngeal regiOn: abundant rhabditOgen glands with xanthOphil secretiOn and cyanOphil glands with amOrphOus secretiOn ( Figs. 8–11), besides sparser xanthOphil glands with cOarse granular secretiOn. Creeping sOle Occupies whOle bOdy width. Glandular margin (Figs. 8,9) discernible after first millimetre Of bOdy. Glands Of at least three typescOnstitute glandular margin: glands with cyanOphil, finely granular secretiOn and glands with erythrOphil cOarse granular secretiOns, bOth abundant, besides sparser rhabditOgen glands with xanthOphil secretiOn. The same glands discharging thrOugh epidermis Of pre-pharyngeal regiOn are present in the anteriOr tip ( Figs. 6, 7). CutaneOus musculature with usual three layers (circular, Oblique and lOngitudinal layers); lOngitudinal layer with thick bundles ( Figs. 8–11, Table 2). Thickness Of cutaneOus musculature between twO and three times that Of epidermis ( Table 2). Ventral and dOrsal musculatures with similar thickness at sagittal plane in pre-pharyngeal regiOn ( Table 2). Thickness Of cutaneOus musculature becOming prOgressively lOwer tOwards bOdy margins and anteriOr tip.   FIGURES 2–5.  Obama allandra  sp. nov., holotype: (2) dorsal pattern of pigmentation; (3) eye pattern; detail of eye pattern and pattern of pigmentation at anterior body region (4) and median third of the body (5). Anterior tip to the left.   FIGURES 6–13.  Obama allandra  sp. nov., holotype, microphotographs of transverse (6–11) and sagittal sections (12–13; anterior tip to the left): (6) anterior region of body; (7) detail of anterior region of body; (8) pre-pharyngeal region; (9) detail of body margin of pre-pharyngeal region; (10) detail of dorsal surface of pre-pharyngeal region; (11) detail of ventral surface of pre-pharyngeal region; (12) pharynx; (13) ovary. The ovary is shown in a parasagittal section in order to show the ovovitelline duct opening.   TABLE 2.Body height and cutaneous musculature in the median region of a transverse section of the pre-pharyngeal region, in micrometers, and ratio of the height of cutaneous musculature to the height of the body (mc:h index) of specimens of  Obama allandra sp. nov.    Holotype Paratype  MZU PL.00284  Dorsal cutaneous musculature 55 65  Ventral cutaneous musculature 50 70  Dorsal epidermis 23 19  Ventral epidermis 23 23  Body height 950 1240  Mc:h (%) 11 11 Mesenchymal musculature ( Figs. 8, 10–11) well develOped, cOmpOsed Of three layers: (1) dOrsal subcutaneOus, lOcated mainly clOse tO cutaneOus musculature, with decussate fibres (6–12 fibres thick), (2) supra-intestinal transverse (5–9 fibres thick) and (3) sub-intestinal transverse (4–7 fibres thick), besides fibres in variOus directiOns. Mesenchymal musculature less develOped in anteriOr regiOn than in pre-pharyngeal regiOn ( Fig. 6).  Pharynx.Pharynx cylindrical, nearly 7% Of bOdy length, Occupies mOst Of the pharyngeal pOuch. DOrsal pharyngeal insertiOn slightly shifted pOsteriOrly regarding the ventral insertiOn. MOuth lOcated in pOsteriOr third Of pharyngeal pOuch ( Fig. 12). OesOphagus shOrt, with fOlded walls. OesOphagus: pharynx ratiO 12–15%.  Reproductive organs.Testes in up tO fOur irregular rOws On either side Of bOdy, lOcated beneath dOrsal transverse mesenchymal muscles, between intestinal branches ( Figs. 8, 10), begin slightly anteriOrly tO Ovaries, in anteriOr third Of bOdy, and extend laterally tO pharynx ( Table 1). Sperm ducts dOrsal tO OvOvitelline ducts, abOve Or amOng fibres Of sub-intestinal transverse mesenchymal musculature, in pre-pharyngeal regiOn ( Fig. 11). Spermiducal vesicles, beginning laterally tO pharynx, enter terminally intO prOximal pOrtiOn Of prOstatic vesicle ( Figs. 14–15). PrOstatic vesicle, lOcated near cOmmOn muscle cOat, with prOximal pOrtiOns fOrked, glObOse Or OvOid, and distal pOrtiOn wide and inverted-U shaped while Outside penis bulb, narrOwing and becOming sinuOus after penetrating cOmmOn muscle cOat. In paratypeMZU PL. 0 0 284, fOrked pOrtiOns shOw asymmetry, with left pOrtiOn mOre elOngate than right pOrtiOn ( Fig. 15). FOrked pOrtiOns with prOximal end clOser tO ventral epidermis than tO dOrsal epidermis ( Figs. 14–16). EjaculatOry duct sinuOus, Opening at tip Of penis papilla in specimens MZU PL. 0 0 283 and MZU PL. 0 0 284 ( Figs. 15, 17, 19), whereas clOse tO ventral insertiOn Of papilla in the hOlOtype( Figs. 14, 20). Male atrium with sOme fOlds, sOmetimes with high and cOmplex fOlding ( Figs. 14–17), and a dOrsO-lateral invaginatiOn. Penis papilla either cOnical, almOst symmetrical, Or with irregular shape and asymmetrical ( Table 1; Figs. 14–17). NumerOus cyanOphil glands with amOrphOus secretiOn, as well as glands with erythrOphil, finely granular secretiOn, Open intO bOth prOstatic vesicle and ejaculatOry duct, the latter mOre numerOus in ejaculatOry duct. Abundant erythrOphil and cyanOphil glands with amOrphOus secretiOn discharge thrOugh the penis papilla and epithelial lining Of male atrium. In additiOn, heavily stained cyanOphil glands, with finely granular secretiOn, Open thrOugh penis papilla and numerOus glands with mixed secretiOn (granules with a cyanOphil peripheral part and an erythrOphil cOre) cOncentrate their Openings thrOugh a dOrsO-lateral invaginatiOn Of male atrium ( Figs 15–18). Muscularis Of the penis papilla and male atrium cOmprised Of mixed circular and lOngitudinal fibres. Vitelline fOllicles ( Figs. 8, 10–11) well develOped in specimens MZU PL. 0 0 283 and MZU PL. 0 0 284, less develOped in the hOlOtype. Ovaries Oval-elOngate, apprOximately fOur times lOnger than wide ( 0.7 mmin its lateral axis), dOrsal tO ventral nerve plate, in anteriOr third Of bOdy ( Fig. 13, Table 1). OvOvitelline ducts emerge dOrsOlaterally frOm median third Of Ovaries ( Fig. 13) and run pOsteriOrly amOng fibres Of sub-intestinal transverse mesenchymal musculature ( Fig. 11). Ascending pOrtiOn Of OvOvitelline ducts lOcated lateral tO female atrium ( Figs. 14, 15). COmmOn glandular OvOvitelline duct shOrt, lOcated dOrsally tO pOsteriOr third Of female atrium ( Figs. 14–16). Female genital duct dOrsO-anteriOrly curved ( Figs. 14–16). Female atrium Oval-elOngate with fOlded walls ( Figs. 14–17), with similar length as that Of male atrium Or sOmewhat lOnger ( Table 1). ParatypeMZU PL. 0 0 283 shOws female atrium larger, partially Occupied by elOngate, cOnical fOld that resembles a penis papilla prOceeding frOm dOrsal wall Of male atrium ( Fig. 17).   FIGURES 14–15.  Obama allandra  sp. nov., (14–15) sagittal composite reconstructions of copulatory apparatus of the holotype (14) and paratype MZU PL. 0 0 284 (15). Anterior tip to the left. The arrow indicates the invagination of the dorsolateral wall of the male atrium receiving numerous openings from glands with mixed secretion. Epithelial lining Of female genital duct and atrium tall cOlumnar, irregular in height (15–90 µm thick), with stratified appearance in sOme parts Of the female atrium ( Fig. 16); epithelial cells with sOme lacunae cOntaining secretiOn. NumerOus cyanOphil glands with amOrphOus secretiOn and scarcer erythrOphil glands with fine granular secretiOn Open intO female duct and atrium. COmmOn muscle cOat thin alOng bOth male and female atria cOmpOsed Of circular, lOngitudinal and Oblique fibres. Male and female atria separated by dOrsal fOld, Obliquely crOssing the atria ( Figs. 14, 17). GOnOduct almOst vertical at sagittal plane ( Figs. 14–17). 1917075575 2015-02-07 MZUSP General On Natural Heritage Private Reserve 2 101 2 Parana holotype