Folia Fac. Sci. Nat. Univ. Masaryk. Brun., Biol. 110 (2007): 53-65

Newsletter on No. 10: Proceedings of the 7 th International Symposium on Enchytraeidae; May 25-28, 2006, Brno, Czech Republic Edited by J. SCHLAGHAMERSKÝ

Fridericia brunensis sp. n. (: Enchytraeidae) – a new European enchytraeid species similar to F. monochaeta Rota, 1995

JI ŘÍ SCHLAGHAMERSKÝ

Masaryk University, Faculty of Science, Department of Botany and Zoology, Kotlá řská 2, 611 37 Brno, Czech Republic; e-mail: [email protected]

ABSTRACT

Fridericia brunensis sp. n. (Clitellata: Enchytraeidae), a species similar to F. monochaeta Rota, 1995 with single dorso-lateral chaetae in most postclitellar segments, preclitellar chylus cells, large seminal vesicle, large sperm funnels, and fused spermathecal ampullae is described based on material from a municipal forest of the city of Brno, Czech Republic. In comparison to its co-geners the species seems very tolerant to acidic and dry conditions. Additional records from Hungary and Germany are discussed.

Keywords: Enchytraeidae, Fridericia, , species description

INTRODUCTION

The newly described species was found during an ecological study of enchytraeids conducted in a municipal forest of the city of Brno (Czech Republic) in 2003-2004 (ŠÍDOVÁ & SCHLAGHAMERSKÝ, 2007). It was originally listed as F. cf. monochaeta due to its similarity with the recently described Fridericia monochaeta Rota, 1995 (SCHLAGHAMERSKÝ, 2007; ŠÍDOVÁ & SCHLAGHAMERSKÝ, 2007). As some characters differed from F. monochaeta , additional specimens were investigated later to clarify the taxonomic status of the Brno population. In a discussion with Prof. Klara Dózsa- 54 JI ŘÍ SCHLAGHAMERSKÝ

Farkas, Dr. Rüdiger M. Schmelz and Dipl.-Biol. Ulfert Graefe following the presentation of a preliminary view on the taxonomic status of this population during the International Symposium on Enchytraeidae in 2006 (SCHLAGHAMERSKÝ, 2006) it became clear that this was indeed a species not yet described and apparently of a wider distribution, at least within Central Europe. The species seems also interesting from the ecological point of view as the localities at Brno are characterised by a dry and acidic soil; at the more extreme of the study sites it was found as one of only two to three enchytraeid species present (ŠÍDOVÁ & SCHLAGHAMERSKÝ, 2007; SCHLAGHAMERSKÝ & ŠÍDOVÁ, 2007). In general, Fridericia species are considered to avoid soils of such acidity (GRAEFE & SCHMELZ, 1999).

MATERIAL AND METHODS

Study sites

The species was found in each of four circular plots (177 m 2 each) that were investigated in a municipal forest of the City of Brno (South Moravia, Czech Republic) situated on the north-west to east-facing slopes south-west of the city quarter Jundrov at an altitude of about 310 m a.s.l. Two plots in close vicinity (49°12'18"N, 16°32'41"E) were within the Holedná Game Preserve and two in the forest adjacent to this enclosure (49°12'11"N, 16°32'48"E; 49°12'16"N, 16°33'3"E). Each pair of plots represented one coniferous (spruce, pine, larch) and one mixed forest stand (with pine, beech, oak and black locust). The farthest distance between plots was ca. 400 m. The soil was illimerized podsol, the soil texture class silt loam, the humus form moder (to mor in the coniferous stands) and soil pH ranged between 3.9 and 4.7 in H 2O and 3.1 and 3.9 in KCl, respectively (range of mean values for each plot is given; the O L/F litter layer of 3 cm height and the underlaying organic and anorganic soil layers to an approximate depth of 12 cm were measured separately; for more detailed soil characteristics see ŠÍDOVÁ & SCHLAGHAMERSKÝ, 2007). Mean annual rainfall in the area is 550 mm and mean annual temperature 8 °C (ČERMÁK, 2003), these values indicate a wetter and colder climate than elsewhere within the city boundaries.

Sampling and investigation of specimens

During the above-mentioned ecological study based on monthly sampling from Dec. 2003 to Dec. 2004 (some preliminary samples were already taken in Nov. 2003) in all four circular plots (6 soil cores per plot), 1833 specimens of F. brunensis sp. n. were identified (as F. cf. monochaeta ). This included many juvenile specimens or those with immature sexual organs (identification was possible due to the characteristic chaetal formula and the low number of Fridericia species present) and the individual specimens were investigated with various intensity. Additional samplings (mostly in one plot only) on Nov. 4, 2005, March 20 Fridericia brunensis sp. n. – a new European enchytraeid species similar to F. monochaeta 55

and May 3, 2006 and Jan. 17, 2007 yielded altogether 135 specimens, 51 of which were subadults with more or less well-developed sexual organs and 16 adults with well-developed sexual organs, including seminal vesicle and clitellum, and mostly also with a yolky egg. All specimens were collected using a cylindrical soil corer of 17 cm 2 surface area and subsequent modified O’Connor wet funnel extraction (12 hours of cold extraction followed by heating of soil samples in 2003-2004; 48 hours of cold extraction without subsequent heating in case of the additional samples taken in 2005-2007). Extracted specimens were stored in Petri dishes with tap water at 5-8 °C prior to identification alive under the microscope. All specimens were investigated alive, all those collected since autumn 2005 using an Olympus BX 51 compound microscope with a magnification of up to 400x (optionally using Nomarski interference contrast). Relevant characters were recorded by digital microscopic photographs, free-hand drawings and notes taken. Four specimens were fixed in hot Bouin’s fluid, stained with paracarmine, and reinvestigated as whole mounts in Canada balsam (SCHMELZ, 2003). These have been deposited at the Natural History Museum (Dept. of Zoology) of the National Museum in Prague. In the following description, “viv” and “fix” refer to observations on living and preserved material, respectively.

RESULTS

Fridericia brunensis sp. n.

Holotype: P6E2996; mature specimen, stained and whole-mounted.

Type locality: Czech Republic, Brno-Jundrov, Holedná Game Preserve (49°12’N, 16°32’E); mixed forest, leg. Ji ří Schlaghamerský, 2007.

Paratypes: P6E2996 (as holotype); three mature specimens, stained and whole-mounted. Same locality as above, leg. Ji ří Schlaghamerský, 2007.

Material examined: 16 adult and 51 subadult specimens were investigated alive in detail, 4 of the adult specimens were whole-mounted and present the type series.

Etymology: Brunensis is the Latin adjective of Bruna, the historic Latin name of Brno (Czech) or Brünn (German), the place from which the species is described and the author’s place of birth.

Description: Medium-sized Fridericia species, length ca. 10 (7-12) mm (viv), diameter 0.25-0.3 mm at XII (viv), segment number 39-50. Chaetae: Formula (0, 1), 2 – (2), 1 : 2, (1) – 2 (1); laterally from XIV-XVII (mostly XIV or XV) to rear end only one chaeta; maximum length of chaetae 40 µm; often floating packages of chaetae in segments VIII-X. 56 JI ŘÍ SCHLAGHAMERSKÝ

Epidermal gland cells present in three to five transverse rows, greyish, not conspicuous (observed in anterior segments). Body wall rather thin (ca. 20 µm). Brain (cerebral ganglion) elongate, ca. 90 µm long and 55 µm wide (fix), sides parallel, anteriorly rounded, posteriorly slightly rounded to truncate. Pharyngeal (“septal”) glands: Three mid-dorsally united pairs in IV-VI (Fig. 1 A). Oesophageal appendages (“peptonephridia”) unbranched, not coiled, with large lumen (particularly in proximal part), rather short - reaching up to VI (Fig. 2). Spermatheca : Ectal duct rather long (ca. 250 µm; width: 16 µm, somewhat wider close to ampulla - viv), with very small, sessile gland at its ectal orifice (Figs 1 A, 1 B, 2); ampulla (including “ental duct”) cylindrical, elongate, ca . 140 µm long and 50 µm wide (viv), with two large, bean-shaped diverticula (ca. 60 µm long and 25 µm wide); ampullae proximally broadly fused at about half their length, with common opening into oesophagus dorsally (Fig. 3). Coelomocytes (Fig. 1 C): Coelomo-mucocytes elipsoid, without refractile vesicles, “type a” (MÖLLER, 1971), ca. 25 µm long; coelomo-lenticytes small (ca. 7 µm). Nephridia (Fig. 1 D): Five pairs in preclitellar segments (VI/VII-X/XI). Alimentary channel with chylus cells in IX-XI (Fig. 4), occupying 1-2 segments, mostly including X; ventral gut endothelium in several posterior segments inflated (XXXV-XL). Dorsal blood vessel rising in XIV-XVI, most often in XV (Fig. 4). Clitellum usually girdle-shaped but in some specimens with very few gland cells ventrally. Seminal vesicle not always present, when present mostly very large (reaching across 2-3 segments), filled with red-brown spermatozoa (Fig. 4). Spermatozoa ca. 400 µm long (viv); heads ca. 140 µm long (viv), reddish brown to iridescent green to black. Sperm funnel large (about 1.3x as long as body width in XII), ca. 350 µm long and 80 µm wide (length : width ≈ 4 : 1), collar slightly narrower than funnel body (Figs 1 E, 5). Vas deferens 7 µm wide (viv). Male copulatory organ small (ca. 100 µm long and 50 µm wide - viv), bursal slit bent (concavity oriented laterally) with short transverse branches (Fig. 1 F). Subneural glands absent but a field of inflated cutaneous cells (somewhat similar to an area glareosa) present mid-ventrally in XIII, located medially behind ventral chaetal bundles (Fig. 6). One mature, yolky egg at a time.

Differential diagnosis: The character combination of (1) a chaetal formula with single chaetae dorso-laterally on most postclitellar segments except the very first ones (XIII, XIV, rarely as far as XVI) and (2) proximally fused spermathecal ampullae, is shared only by two valid species of the known at present, i.e. F. connata Bretscher, 1902 and F. monochaeta Rota, 1995. F. connata is a much larger species (14-25 mm long, up to 70 segments), its chaetae are larger and stouter than in F. brunensis and also differ by their abrupt tapering. In contrast to F. brunensis , chylus cells are present in postclitellar segments and the dorsal vessel starts much more posteriorly. The epithelium of spermathecal ampulla has a distinct warty structure in F. connata not found in F. brunensis , the diverticula differ in shape, being more like converging fingers oriented distad. F. monochaeta is more similar to F. brunensis by its comparable size, being even slightly smaller (up to 44 segments). The reported maximum length of chaetae (55 µm) exceeds that observed in F. brunensis . Fridericia brunensis sp. n. – a new European enchytraeid species similar to F. monochaeta 57

ce

pp B

oe C

D

pg IV

st E

pg V

pg VI - vl

pg VI - dl

F A

200 µm Figure 1: Fridericia brunensis sp. n.: A – anterior body region (based on whole mounted holotype; scale see scale bar): ce – cerebral ganglion, pp – pharyngeal pad, oe – oesophageal appendage, st – spermatheca, pg – pharyngeal gland, vl – ventral lobe, dl – dorsal lobe, IV-VI – segment numbers; B – spermathecae; C – coelomocytes; D – preclitellar nephridium; E – male copulatory organ with bursal slit; F – sperm funnel (B-F based on drawings of microscoped live specimens, not to scale). 58 JI ŘÍ SCHLAGHAMERSKÝ

In difference to F. monochaeta , three chaetae per bundle were never observed in F. brunensis . The sperm funnel of F. monochaeta is much smaller and less elongate than that of F. brunensis, a seminal vesicle has not been reported. The body cavity of F. monochaeta is filled with many coelomocytes, this has not been observed in F. brunensis . In contrast to F. monochaeta , F. brunensis has no conspicuously large male copulatory organ with an area glareosa lining the bursal slit. F. monochaeta lacks the field of inflated epidermal cells mid-ventrally in XIII present in F. brunensis . Spermathaecae of both species seem very similar, a minute ectal gland has not been reported in F. monochaeta by ROTA (1995) but was observed in otherwise matching populations by SCHMELZ (2003); the ectal duct is of similar length but less thin than in F. monochaeta , the diverticula are larger and more bean or kidney-shaped than spherical as in F. monochaeta .

DISCUSSION

DÓZSA-FARKAS (2007) found Fridericia specimens most probably belonging to the same species during faunistic sampling in the Zemplén Mountains in northeast Hungary and also reported them as F. monochaeta. Her notes (Dózsa-Farkas, in lit.) based on the investigation of live material are widely consistent with my observations on the Brno population, although the Hungarian specimens were somewhat longer: number of segments (36)-44-51; body length 11-15 mm; type a oesophageal appendages; 5 preclitellar pairs of nephridia; coelomo-mucocytes (type a) 28-38 µm, coelomo-lenticytes 7-12 µm; chylus cells in VIII-X, IX-X or X-XI; dorsal vessel originating in XIV or XV; sperm funnel (230)-300- 370 µm, 2-3.7x as long as wide, as long or longer as body diameter; sperm heads 140 µm long, entire spermatozoa ca. 400 µm long; large seminal vesicle; small gland at the orifice of spermathecal ectal duct, spermathecal diverticula somewhat larger and more elongate as in the drawing of F. monochaeta by ROTA (1995); subneural gland in XIII. With regard to the last character, I have originally also considered the inflated epidermal cells mid-ventrally (i.e. above the ventral nerve chord) on XIII as the sign of the presence of a subneural gland. However, R. Schmelz (pers. com.) drew my attention to the fact that the field of inflated cells present in F. brunensis does not fully resemble a typical area glareosa above a subneural gland and that there is actually no gland attached to the nerve chord in this position. Such a glandular thickening of the epidermis in a ventral position more or less close behind the clitellum was observed in several Fridericia species (SCHMELZ, 2003) . Based on the investigation of live specimens and microphotographs of F. brunensis , both from the type locality, U. Graefe (pers. com.) reported to have found the same species in the lawn of an urban park in Hamburg, Germany (soil monitoring plot in the Amsinckpark) in 2002. At that time he recognized it as new Fridericia species, listed it as “ Fridericia sp. n. (mono)” in an unpublished report (GRAEFE et al., 2003), but did not describe it. Whereas at Brno F. brunensis was a dominant constituent of very poor to rather poor assemblages (2-9 enchytraeid species per plot), the population at the Amsinckpark was part of a species-rich enchytraeid Fridericia brunensis sp. n. – a new European enchytraeid species similar to F. monochaeta 59

assemblage of 43 species (including species preferring strongly acidic soils as Marionina clavata Nielsen & Christensen, 1961, Cognettia sphagnetorum (Vejdovský, 1877) and Achaeta brevivasa Graefe, 1980 but also many species preferring neutral soils, for instance, Stercutus niveus Michaelsen, 1888 and 10 additional Fridericia species). At the same time it reached the second highest dominance (12 %) of all enchytraeid species present in the Amsinckpark plot, including Hrabeiella periglandulata (“Polychaeta”), a remarkable tiny that was also present in most of the Brno study plots. The pH (CaCl 2) range of soil (0-16 cm) in the Amsinckpark plot was 4-5 (GRÖNGROFT et al., 2003). It is thus somewhat surprising that GRAEFE et al. (2003) assigned the species a “reaction figure” of 7, corresponding to an indicator of slightly acidic to slightly alkaline soils and identical to that of most other Fridercia species classified. In light of the conditions at the Brno localities, F. brunensis seems remarkably tolerant to acidic conditions as well as to drought. Assuming that the populations from northern Germany and northeastern Hungary belong to F. brunensis sp. n., this species has at least a Central European distribution. It is therefore somewhat surprising that it has not been described up to now. As the most similar species, F. monochaeta, has also been described rather recently (ROTA, 1995), some earlier records of F. connata might actually be based on finds of F. brunensis , particularly as the taxonomic value of the position of chylus cells has been disregarded for a long time (SCHMELZ, 2003).

ACKNOWLEDGEMENTS

The study was conducted under the Masaryk University’s Research Plans MSM 143100010 (in 2003-2004) and MSM 0021622416 (in 2005-2007). My student, Adéla Šídová, originally drew my attention to the taxonomic problem posed by specimens of this species and was instrumental in sampling, extracting and sorting during the fist phase of the study. Ms. Marcela R ůži čková assisted with producing the whole mounts. I am also grateful to the Institute of Soil Biology (Academy of Sciences of the Czech Republic) for chemical and physical analyses of soil and to the Brno Municipal Forest Administration (Lesy m ěsta Brna) for information about the game preserve.

REFERENCES

ČERMÁK, K., 2003. Studie obora Holedná [Study on the Holedná Game Preserve]. Brno, 19 pp. (manuscript; in Czech). DÓZSA-FARKAS, K., 2007. Comparative enchytraeid faunistic investigation of the northeastern mountain range in Hungary. In: TAJOVSKÝ, K., SCHLAGHAMERSKÝ, J. & PIŽL, V. (eds), Contributions to Soil Zoology in Central Europe II. ISB BC AS CR, v.v.i., České Bud ějovice, pp. 29-34. 60 JI ŘÍ SCHLAGHAMERSKÝ

GRAEFE, U. & SCHMELZ, R.M. 1999. Indicator values, strategy types and life forms of terrestrial Enchytraeidae and other microannelids. Newsletter on Enchytraeidae, 6: 59-67. GRAEFE, U., BUSCH, W. & EGBERTS, B., 2003. Bodenzoologische Untersuchungen auf der Boden-Dauerbeobachtungsfläche Amsinckpark – Untersuchungsjahr 2002. Institut für Angewandte Bodenbiologie GmbH, Hamburg (unpublished report to the environmental authorities of the City of Hamburg – Freie und Hansestadt Hamburg), 28 pp. GRÖNGROFT, A., HOYER, P. & SCHUBERT, L., 2003. Wiederholungsuntersuchung an der Boden-Dauerbeobachtungsfläche Amsinckpark. Universität Hamburg, Fachbereich Geowissenschaften (unpublished report to the environmental authorities of the city of Hamburg – Freie und Hansestadt Hamburg), 23 pp. ROTA, E., 1995. Italian Enchytraeidae (Oligochaeta). Boll. Zool., 62: 183-231. SCHMELZ, R.M., 2003. Taxonomy of Fridericia (Oligochaeta, Enchytraeidae). Revision of species with morphological and biochemical methods. Abhandlungen des Natur- wissenschaftlichen Vereins in Hamburg (Neue Folge), 38: 1-488. SCHLAGHAMERSKÝ, J., 2006. Fridericia monochaeta brunensis spp. n. – a northern or eastern subspecies? In: SCHLAGHAMERSKÝ, J. (ed.), 7 th International Symposium on Enchytraeidae, Brno, Czech Republic, May 25-28, 2006, Abstract Book with programme and list of participants. Masaryk University, Faculty of Science, Institute of Botany and Zoology, Brno, p. 23. SCHLAGHAMERSKÝ, J., 2007: Consequences in the advance in Fridericia taxonomy for our knowledge of Czech and Slovak enchytraeid faunas. In: TAJOVSKÝ, K., SCHLAGHAMERSKÝ, J. & PIŽL, V. (eds), Contributions to Soil Zoology in Central Europe II, ISB BC AS CR, v.v.i., České Bud ějovice, pp. 127-130. SCHLAGHAMERSKÝ, J. & ŠÍDOVÁ, A., 2007. On a species-poor enchytraeid community of peculiar composition. In: SCHLAGHAMERSKÝ, J. (ed.) Newsletter on Enchytraeidae No. 10: Proceedings of the 7th International Symposium on Enchytraeidae, May 25-28, 2006, Brno, Czech Republic. Folia Fac. sci. nat. Univ. Masaryk. Brun., Biologia, 110: 183-192. ŠÍDOVÁ, A. & SCHLAGHAMERSKÝ, J., 2007. The impact of high game density on enchytraeids in a mixed forest. In: TAJOVSKÝ, K., SCHLAGHAMERSKÝ, J. & PIŽL (eds), V. Contributions to Soil Zoology in Central Europe II. ISB BC AS CR, v.v.i., České Bud ějovice, pp. 147-152.

Fridericia brunensis sp. n. – a new European enchytraeid species similar to F. monochaeta 61

Figure 2: Fridericia brunensis sp. n.: Oesophageal Appendage (OesApp) and spermathecal ectal duct with minute gland at its orifice (EctGl) indicated by arrows; spermathecal ampullae visible below focused level (microscopical photograph from live specimen).

Figure 3: Fridericia brunensis sp. n.: Fused spermathecal ampullae with diverticula (microscopical photograph from live specimen) 62 JI ŘÍ SCHLAGHAMERSKÝ

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Fridericia brunensis sp. n. – a new European enchytraeid species similar to F. monochaeta 63

Figure 4: Fridericia brunensis sp. n.: Clitellar region (IX/X-XIII/XIV) with chylus cells (Chylus), large seminal vesicle (SemVes) and origin of dorsal vesel (DV) indicated by arrows (microscopical photograph from live specimen). 64 JI ŘÍ SCHLAGHAMERSKÝ

prázdná Fridericia brunensis sp. n. – a new European enchytraeid species similar to F. monochaeta 65

Figure 5: Fridericia brunensis sp. n.: Sperm funnels with attached spermatozoa (microscopical photograph from live specimen).

Figure 6: Fridericia brunensis sp. n.: Inflated cutaneous cells mid-ventrally in posterior part of XIII (microscopical photograph from live specimen). 66 JI ŘÍ SCHLAGHAMERSKÝ

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