Three new Fridericia species (Oligochaeta, Enchytraeidae) from Transylvania, Romania Author Boros, Gergely Author Dózsa-Farkas, Klára text Zootaxa 2015 3911 3 357 368 journal article 42337 10.11646/zootaxa.3911.3.3 d32cda60-d7ba-428f-b532-414fe63d0212 1175-5326 239353 744769AD-E4D6-4AD3-A067-4006095378F5 Fridericia transylvanica sp. n. ( Figs 2 A–E, 3, 4) Type material. Holotype : F.19, slide No. 1012. Type locality: N45.89052 E24.78019 , 580 m asl, near the village of Cincu, Romania . Leg. G. Boros, 15.10.2013 . Adult whole mount stained with borax carmine. Paratypes : P. 96.1–9, slides No. 1027, 1028, 1033, 1050, 1056–1060 , 9 adult whole mounts. N45.89052 E24.78019 , 580 m asl, near the village of Cincu, Romania . Leg. G. Boros, 15.10.2013 . (No. 1027, 1028, 1033, 1050 stained with bromophenol blue, No. 1056–1060 without staining.) P.96.10–13, 3 adults and 1 subadult specimen in 70 % ethanol. (Unfortunately, the fixed specimens dried out after investigation.) N47.68111 E23.78722 , 698 m asl, Mogosa forest, Gutin Mts., Romania . Leg. K. Dózsa-Farkas, 17.0 9.2005. P. 96.14–18, slides No. 349, 1052–1055 , 3 adult specimens and 2 subadults . N44.62987 E22.45085 , 432 m asl, near Petrovo Selo, Djerdap National Park, Serbia . Leg. L. Dányi, J. Kontschán and Zs. Újvári, 28.10.2010 . No. 349, 352–353 stained with borax carmine, No. 1052–1055 without staining. Etymology. Named after the region of the type locality. Diagnosis. The new species can be recognized by the following combination of characters: (1) large size ( 16–26 mm in vivo ), segments 52–69; (2) maximum 4 chaetae per bundle; (3) clitellum girdle-shaped: hyalocytes and granulocytes arranged in transverse rows dorsally, between the mail pores granulocytes only; (4) five preclitellar pairs of nephridia; (5) coelomo-mucocytes c-type, lenticytes scarce; (6) chylus cells mostly in XIII–XIV; (7) bursal slit T-shaped; (8) very large seminal vesicle; (9) subneural glands in XIII, XIV; (10) sperm funnel cylindrical, approximately half as long as body diameter, collar narrower as funnel body, sperm long, sperm heads 160–200 µm long; (11) spermathecae separate entally, with two sac-like diverticula bent distad and forming a common U-shaped sperm-containing chamber together with ampulla. Description. Holotype in vivo 22.5 mm long, 380 µm wide at VIII and 490 µm at the clitellum ( 17.5 mm long, 460 µm wide at VIII and 560 µm at the clitellum, fixed), 60 segments. Body length of the paratypes 15.5–26.5 mm , average width 471±55 µm at VIII and 564±58 µm at the clitellum ( in vivo ). Length of fixed specimens 12–24 mm , average width 479±50 µm at VIII and 586±60 µm at the clitellum. Segments 52–69. The Serbian specimens, except two, somewhat smaller, probably not fully mature. Chaetal formula: 1,2,3,4 – 4,3,2,(0,1): 2,3,4 – 4,3,2. As in other Fridericia species, chaetae in bundles of 4 arranged in pairs with the outer pair being longer and thicker than the inner pair: 65– 70 x 5 Μm against 30– 40 x 3–4 Μm (preclitellar bundles); chaetae in bundles of 2 or 3 mostly unequally long; chaetae smaller in mid-part of body, maximum length about 40–50 Μm, from about XXVII on increasing again in size; from XXX–XXXII only two equally long chaetae per bundle (sometimes only one or absent), length increasing to about 75– 85 x 6–7 Μm at body-end. Head pore at 0/I. Dorsal pores from VII. Epidermal gland cells arranged in 2–3 transverse rows per segment. Clitellum in XII–1 /2XIII, girdle-shaped, large hyalocytes (diameter 20–28 Μm) and granulocytes (diameter 14–18 Μm), arranged in rows dorsally ( Fig. 3 A), between and near the bursal slits only granulocytes ( Fig. 3 B,C). Thickness of body wall about 30–45 µm, cuticle about 3–5 µm in fixed specimens. Brain egg-shaped, about 220 Μm long (fixed) and 2 times longer than wide. FIGURE 2. Fridericia transylvanica sp. n. (A–E): A .Nephridium, B . C oelomocytes. C . Sperm funnel. D . Spermatheca. E . Oesophageal appendage. Fridericia gyromonodactyla sp. n. (F–I): F . Pharyngeal gland. G . S perm funnel. H . Coelomocytes. I . Spermathecae. Fridericia longeaurita sp. n. (J–N): J . Spermathecae. K . Single spermatheca (fusion with the other not observed). L. Sperm funnel, M. Coelomocytes, N . Oesophageal appendage. FIGURE 3. Micrographs of Fridericia transylvanica sp.n. A. Citellar glands dorsally in vivo , B. Clitellar glands laterally, ventral side to the top (unstained whole mount), C. Clitellum ventrally in vivo (glands weakly developed, ventrally only granulocytes), bursal slits marked with arrows. D . Typical T-shaped bursal slit in vivo , marked with arrow, E . Coelomomucocytes fixed and stained. F . Male copulatory apparatus marked with white arrows, chylus cells marked with black arrow (unstained whole mount), G . Nephridium in IX/X, H . Sperm funnels in vivo (marked with arrow), I . Intestinal contents with many crystals inside ( in vivo ). All scale bars except in 3I = 50 Μm. FIGURE 4. Micrographs of Fridericia transylvanica sp.n. A . Spermathecae, fixed and stained (ph= pharyngeal glands). B . One spermatheca in vivo , the long spermathecal duct with the large brown ectal gland (marked with arrow) clearly visible. C . Spermathecal ampullae with the two diverticula, in vivo . D . Fixed and stained spermatheca (sperm in diverticula). E . Subneural gland in XIV (fixed and stained), marked with arrow. Scale bars = 50 Μm. Oesophageal appendages a-type, long, with 2–3 short terminal branches ( Fig. 2 E). Pharyngeal glands all united dorsally, in IV without and in V and VI with ventral lobes. Chloragocytes from V, about 32–36 Μm long, after XIV–XV smaller, brown in vivo. In the intestine many soil crystals can be found ( Fig. 3 I). Dorsal vessel from XVII–XIX, blood colourless. Midgut pars tumida in XXVII–XLIV occupying 6–11 segments mostly 8–9 segments. Five pairs of preclitellar nephridia ( Fig. 2 A, 3G) from 6/7 to 10/11 , length ratio anteseptale: postseptale 1:1.8–2, subterminal origin of efferent duct. Coelomo-mucocytes c-type (length 30–70 Μm in vivo , 20–60 µm fixed), lenticytes scarce, 8–10 Μm long ( Fig. 2 B, 3E). Chylus cells between XIII–XV, but often not visible, occupying 2 segments. Seminal vesicle large in X–XII or XI–XIII. Sperm funnels cylindrical ( Fig. 2 C, 3H), about 333–375 µm long and 2–2.5 times as long as wide ( in vivo ). Funnel length in fixed specimens 250–400 µm. Collar narrower than funnel. Spermatozoa long, exact length in seminal vesicle often not measurable, heads 140–160 µm in fixed specimens (160–200 µm in vivo ). Diameter of sperm ducts 7–10 µm (fixed). Male copulatory organs ( Fig. 3 F) 180–240 µm long, 130–190 µm wide and 95–130 Μm high ( in vivo ), bursal slits T-shaped ( Fig. 3 D) but sometimes somewhat divergent ( Fig. 3 C). Large subneural glands in XIII and XIV, the latter is larger ( Fig. 4 E). Spermathecae (Fig.. 2D, 4A–D): one large, stalked ectal gland at the orifice ( Fig. 4 B), length about 100 µm in vivo (80 µm, fixed). In one case there was a smaller second gland (50 µm long). Ectal duct about 470–510 µm long and 28–35 µm wide, projecting into ampulla, large ental bulb about 70–78 µm wide in vivo (45–63 µm, fixed), canal not widened. Each ampulla with two sac-like diverticula bent distad; diverticula and distal part of ampulla forming common U-shaped sperm-containing chamber ( Fig. 4 C). Average width of spermathecae 137±12 µm ( in vivo ), 102±22 µm (fixed). Proximal part of ampulla considerably set off from distal part by a constriction, separate openings into oesophagus. One mature egg at a time. Distribution and habitat. In Romania : Transylvanian Plateau, close to Cincu village. Pastures, meadows. Mogosa forest, Gutin Mts. Beech forest. In Serbia : Serbian Carpathians, Djerdap National Park, close to Petrovo Selo, beech forest. Differential diagnosis. The shape of spermathecae in F. transylvanica sp. n. is very similar to the spermathecae of F. maculata Issel, 1905 and F. maculatiformis Dózsa-Farkas, 1972 , but these species are smaller (36–48 segments, 9–10 mm long) and have a maximum of only two chaetae in a bundle. F. paroniana Issel, 1904 , F. granosa Schmelz, 2003 , F. lenta Schmelz, 2003 , F. discifera Healy, 1975 , F. nix Rota, 1995 , F. chongquingensis Xie, Liang & Wang, 1999 , F. sardorum Cognetti, 1901 and F. montafonensis Schmelz, 2003 have also somewhat similar spermathaecae but the main differences between these species and the new species are as follows: F. paroniana is smaller (40–48 segments, 6–12 mm ) and has only two chaetae per bundle, similar to F. l en t a , which has more segments (50–68) but is only 10–12 mm long and without subneural glands. Subneural glands are absent also in F. n i x , F. granosa , F. chongquingensis and F. s a rd or u m ; in addition F. ni x and F. granosa have characteristically dark coelomo-mucocytes and in the latter the clitellum is saddle-shaped. In F. chongquingensis the maximum number of chaetae per bundle is 5–6 and the oesophageal appendages are of c-type, i.e. much branched. F. sardorum has also more chaetae (6–8), coelomo-mucocytes a-type, and two large brown ectal glands at the spermathecal ectal orifices. F. discifera has subneural glands in XIII and XIV, but the worm is smaller (41–48 segments, 6–13 mm ), the mucocytes are b-type, the spermathecal ectal duct is short, the ectal gland small, and there are only four pairs of preclitellar nephridia. Lastly F. montafonensis has more chaetae per bundle (4–6) and the two spermathecae are fused proximally ( Dózsa-Farkas 2009 ; Schmelz 2003 ).