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Invertebrate Zoology, 2010, 7(1): 2946 © INVERTEBRATE ZOOLOGY, 2010 Proales tillyensis sp.n. (Monogononta: Proalidae), a new rotifer species from North-West Germany, with reconstruction of its somatic musculature E.F. Wilts, W.H. Ahlrichs Systematics and Evolutionary Biology, Department of Biology and Environmental Sciences, Carl von Ossietzky University Oldenburg, 26111 Oldenburg, Germany. e-mail: [email protected] ABSTRACT: We here describe a new proalid rotifer species Proales tillyensis sp.n. from Oldenburg, North-West Germany. The species was initially found in the ph-neutral water of Lake Tilly in August 2006. Its description is based on light and electron microscopy, providing different views of both the whole specimen and its trophi. Additionally, the body musculature of the species was visualized by confocal laser scanning microscopy using fluorescent-labelled phalloidin and compared with the musculature of other Proales species. Proales tillyensis sp.n. resembles P. fallaciosa Wulfert, 1937 and P. decipiens (Ehrenberg, 1832) but differs in its ecology. Furthermore it can be diagnosed from the former species by the absence of a knob-like projection between the toes, the small body size, the number of uncus teeth and the organization of body musculature. From the latter species it can be diagnosed by the number of uncus teeth, the small body size and the lack of a constriction between stomach and intestine. KEY WORDS: Proales tillyensis n.sp., Rotifera, Proalidae, CLSM, somatic musculature. Proales tillyensis sp.n. (Monogononta: Proalidae): íîâûé âèä êîëîâðàòîê èç ñåâåðî-çàïàäíîé Ãåðìàíèè ñ ðåêîíñòðóêöèåé ñîìàòè÷åñêîé ìóñêóëàòóðû Å.Ô. Âèëö, Â.Ã. Àëðèõñ Systematics and Evolutionary Biology, Department of Biology and Environmental Sciences, Carl von Ossietzky University Oldenburg, 26111 Oldenburg, Germany. e-mail: [email protected] ÐÅÇÞÌÅ: Îïèñàí íîâûé âèä ïðîàëèäíîé êîëîâðàòêè Proales tillyensis sp.n. èç Îëüäåíáóðãà (ñåâåðî-çàïàäíàÿ Ãåðìàíèÿ). Âèä áûë îáíàðóæåí â pH íåéòðàëüíîé âîäå â îçåðå Òèëëè â àâãóñòå 2006 ã. Îñîáè áûëè èçó÷åíû ìåòîäàìè ñâåòîâîé è ýëåêòðîííîé ìèêðîñêîïèè. Áûëè ïîëó÷åíû ôîòîãðàôèè öåëûõ æèâîòíûõ è èõ ðîòî- âûõ ÷àñòåé. Äîïîëíèòåëüíî áûëî ïðîâåäåíî èçó÷åíèå ñîìàòè÷åñêîé ìóñêóëàòóðû íîâîãî âèäà ñ ïîìîùüþ ôëþîðèñöåíòíîé ôàëëîèäèíîâîé ìåòêè è ïðîâåäåíî ñðàâ- íåíèå ñ îðãàíèçàöèåé ìóñêóëàòóðû ó äðóãèõ êîëîâðàòîê ðîäà Proales. Proales tillyensis sp.n. ñõîäåí ñ P. fallaciosa Wulfert, 1937 è P. decipiens (Ehrenberg, 1832), íî èõ ýêîëîãèÿ ðàçëè÷íà. Êðîìå òîãî, îò P. fallaciosa íîâûé âèä îòëè÷àåòñÿ îòñóòñòâèåì áîðîäàâêîîáðàçíîãî âûðîñòà ìåæäó ïàëüöàìè, ìåëêèìè ðàçìåðàìè òåëà, èíûì 30 E.F. Wilts, W.H. Ahlrichs ÷èñëîì çóáîâ óíêóñà è èíîé îðãàíèçàöèÿ ñîìàòè÷åñêîé ìóñêóëàòóðû. Îò P. decipiens íîâûé âèä îòëè÷àåòñÿ ÷èñëîì çóáîâ óíêóñà, ìåëêèìè ðàçìåðàìè òåëà è îòñóòñòâèåì ïåðåòÿæêè ìåæäó æåëóäêîì è êèøêîé. ÊËÞ×ÅÂÛÅ ÑËÎÂÀ: Proales tillyensis sp.n, êîëîâðàòêè, Proalidae, êîíôîêàëüíàÿ ëàçåðíàÿ ñêàíèðóþùàÿ ìèêðîñêîïèÿ, ñîìàòè÷åñêàÿ ìóñêóëàòóðà. Introduction Material and methods The monogonont rotifer taxon Proalidae, Samples were collected during August 2006, Proales Gosse, 1886 comprises a genus with 2008 and 2009 from Lake Tilly in Oldenburg, currently 44 species (Segers, 2007). From a sys- north-western Germany (53°04¢35.0²N, 7°12¢ tematic point of view Proales is problematic and 00.07²E) with a plankton sieve of 64 µm mesh in need of a revision because it represents an size. Single rotifer specimens were studied by assemblage of diverse species from which sever- both differential interference contrast light mi- al have been moved in and out in the past (Segers, croscopy (Leica DMLB) and scanning electron 1995; Wilts et al., 2009a; De Smet, in press.). microscopy (Zeiss DSM 940). Light micro- Koste (1978) already subdivided the species of scopic images were taken with a digital camera Proales into two main groups: Group A (group (Olympus ColorView). Isolated rotifer speci- Proales sensu stricto) comprises freshwater spe- mens were narcotized with an aqueous solution cies that resemble the type species P. decipiens of 0.25% bupivacaine (Bucain®) and fixed with and are characterized by a tubular habitus with a 4% OsO in 0.1 M sodium cacodylate buffer and 4 trunk comprising multiple pseudosegments, a picric acid formaldehyde at 240 mOsm (after hardly offset head with a rostrum and a short foot, Melone, 1998). Specimens were dehydrated in a cerebral eye and an elongated buccal field. The a graded ethanol series followed by critical- other group comprises almost unexceptionally point drying. Dried specimens were mounted on marine species resembling Proales reinhardti stubs. For trophi preparations, individual spec- (group B, misleadingly designated as group Proa- imens were dissolved under a stereo micro- les sensu stricto by Wilts et al., 2009a). The scope (Leica MZ12 ) following the procedure 5 representatives of this group are characterized by given by Kleinow et al. (1990) leaving only the a bulbous trunk with an offset head and a long cuticularized trophi elements. Trophi were rinsed foot, apical eyes and two epidermal projections and dried on a coverslip and mounted on stubs. restricting the apical corona caudally. Specimens and trophi were coated with gold We found a new species in a lake near and studied by scanning electron microscopy. Oldenburg, North-West Germany occuring in For CLSM, specimens were anaesthetized with floating gras material during warm periods of bupivacaine, fixed for 1 h in phosphate-buff- the summer months. The new species fits group ered 4% paraformaldehyde, rinsed in 0.1 M A, resembling P. fallaciosa and P. decipiens. In phosphate-buffered saline (PBS), and then made the following we describe the species carefully permeable for staining by exposure to 0.1% on the basis of light and scanning electron Triton X-100 buffered in 0.1 M PBS for 1 h. For microscopic (SEM) findings, and give standar- staining, 2 ml of 38 mM methanolic TRITC- dised views of the specimen and its trophi. labelled phalloidin solution were added to 100 Furthermore, we analyze the somatic muscula- ml of Triton X-100 buffered in 0.1 M PBS. ture of the species using confocal laser scanning Specimens were stained for 3 h and mounted in miscoscopy (CLSM) in order to provide a broad- Citifluor® on a coverslip; a total of five speci- er overview of the variation of the muscular mens were analyzed. The images were obtained pattern across Proales. under a wavelength of 488 nm using a Leica Proales tillyensis sp.n. (Monogononta: Proalidae) 31 TCS SP 5 confocal laser scanning microscope. ed from each other and the trunk by transverse We used Leica LAS AF 1.7.0 for the analysis of folds (Figs. 1A, B, 4A, B, 5A). The anterior the image stacks. All line drawings were pre- pseudosegment carries a small, rounded ros- pared with Adobe Photoshop® CS2. trum. The rostrum is separated from the anterior pseudosegment via a transverse fold. Dorsally Order Ploima Hudson & Gosse, 1886 the pseudosegment has a dorsal antenna (Figs. 1B, 4B, 5A, C) and is followed caudally by the Family Proalidae Bartos, 1959 shorter neck pseudosegment (nps) (Figs. 1A, B, 4A, 5A). The corona appears semicircular in Genus Proales Gosse, 1886 dorsal view. Its long, locomotory cilia are locat- ed apically (Figs. 1B, 5C); the ventral buccal Proales tillyensis sp.n. field consists of numerous short cilia and ex- tends to the center of the neck pseudosegment MATERIAL. Holotype. A parthenogenetic female in (Figs. 4C, 5B). The cylindrical trunk is usually a permanent, glycerin glass slide mount deposited at divided into three pseudosegments differing in Museum fur Naturkunde, Germany, Berlin (ZMB) Gener- size and separated from each other by distinct alkatalog freilebende Wurmer ZMB Vermes 11372. Paratypes. Three parthenogenetic females in perma- transverse folds (Figs. 1A, B, 4A, B, C, 5A). nent, glycerin glass slide mounts (ZMB Vermes 11373-1, The anteriormost, longest and broadest trunk -2 and -3), ten parthenogenetic females mounted on a SEM pseudosegment bears at its posterior third two stub (ZMB Vermes 11373-4, -5, -6, -7, -8, -9, 10, -11, -12 and -13) and a trophi preparation with three trophi mount- antennae in dorsolateral position (Figs. 4A, 5A). ed on a SEM stub (ZMB Vermes 11373-14, -15 and -16). This pseudosegment bears also seven longitudi- TYPE LOCALITY. Lake Tilly in Olden- nal folds (one long dorsomedian fold and three burg, Lower Saxony, Germany. August 2006, long dorsolateral ones on each side) and two 2008, 2009 (53°04¢35.02²N, 7°12¢00.07²E). shorter lateral folds (one on each side) (Figs. ETYMOLOGY. The species name tillyen- 1A, B, 4A, B, 5A). The pseudosegment is fol- sis is derived from the type locality Lake Tilly in lowed by a very short lumbar pseudosegment Oldenburg. (lps). The lumbar pseudosegment is subsequently DIAGNOSIS. Species with long, slender, followed by a preanal pseudosegment (pps) that tubular body; head and foot hardly offset; trunk slightly overlaps the foot (Figs. 1A, B, 4A, 5A). with seven longitudinal folds; apical corona Depending on the degree of contraction of the with elongated, ventral buccal field; foot with animal, the trunk can have two additional trans- two pseudosegments and acute, conical toes; verse folds: One fold in the anterior region (see rami with anterior projections; unci with six arrow Fig. 5A) and one in the posterior region of teeth, largest tooth ventrally with accessory the largest trunk pseudosegment (see arrows toothlet; fulcrum rod-shaped, distally broaden- Figs. 4C, 5A). The completely contractible, ing; manubria club-shaped, slightly curved dis- broad and only slightly tapering foot is divided tally; hypopharynx with paired plates ventrally, into two short foot pseudosegments and two bearing two large teeth each. short, acute, conical toes (Figs. 1A, B, 2D, 4B, DESCRIPTION. Habitus. (Figs. 1A, B, 2A C). Viewed dorsally, the toes are usually spread D, 4AC, 5AC) Proales tillyensis sp.n. has a (Figs. 2D, 5A).