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PONDER Australian Museum Sydney OIE Academia Sinica Qingdao, People's Republic of China D. G. REID The Natural History Museum London, United Kingdom S. G. SEGERSTRÁLE Institute of Marine Research Helsinki, Finland A. STANCZYKOWSKA Siedlce, Poland F. STARMUHLNER Zoologisches Institut der Universitat Wien, Austria Y. |. STAROBOGATOV Zoological Institute St. Petersburg, Russia J. STUARDO Universidad de Chile Valparaiso C. THIRIOT University P. et M. Curie Villefranche-sur-Mer, France thiriot @ obs-vlfr. fr S. TILLIER Museum National d'Histoire Naturelle Paris, France J.A.M. VAN DEN BIGGELAAR University of Utrecht The Netherlands М. Н. VERDONK Rijksuniversiteit Utrecht, Netherlands H. WAGELE Ruhr-Universitát Bochum Germany Heike. Waegele @ ruhr-uni-bochum.de ANDERS WAREN Swedish Museum of Natural History Stockholm, Sweden B. R. WILSON Dept. Conservation and Land Management Kallaroo, Western Australia H. ZEISSLER Leipzig, Germany A. ZILCH Forschungsinstitut Senckenberg Frankfurt am Main, Germany MALACOLOGIA, 2001, 43(1-2): 1-11 GENETIC VARIATION IN THE LAND SNAIL ISOGNOMOSTOMA ISOGNOMOSTOMA (GASTROPODA: PULMONATA: HELICIDAE) P. Van Riel,'? К. Jordaens,? J. |. Van Goethem & T. Backeljau' ABSTRACT Genetic and phenotypic variation in 13 populations of /sognomostoma isognomostoma (Schroter, 1784) from central and peripheral areas of its distribution range in central Europe was estimated by means of allozyme electrophoresis at 11 loci and a morphometric analysis of six shell and 12 genital tract measurements. Because of the effects of genetic drift and inbreeding, which may lead to rapid genetic differ- entiation, peripheral isolates may play an important role in diversification and speciation. Besides this, these populations may also be vulnerable to extinction because of the genetic impoverish- ment resulting from the same population genetic processes. Population differentiation as evidenced by allozyme electrophoresis was very high (mean Ест: 0.550) and genetic variation was found to be partitioned among regions (mean F,-: 0.480), rather than between populations within regions (mean F.,: 0.152). Only low allozyme variation could be detected within local populations (mean F¿: 0.373; mean number of alleles per locus: 1.27; mean expected heterozygosity: 0.062). Although morphological differentiation was less pronounced among populations and/or regions, it revealed mainly the same pattern of variation. No genetic variation, or the loss of it, could be attributed to the position of populations near the periphery of the distribution range. Key words: allozymes, morphology, population genetics, peripheral populations, /sogno- mostoma isognomostoma. INTRODUCTION Isognomostoma isognomostoma (Schróter, 1784) is a terrestrial helicid snail with a distri- bution in Europe extending over the moun- tainous regions of central Europe, from the Harz, Sauerland and the Eifel region in the north, to the Carpathians in the east and the Alps, Jura and Vosges in the south and southwest (Ant, 1963; Kerney et al., 1983). The species is considered to be of Alpine- Carpathian origin and probably dispersed northward during inter- and postglacial peri- ods (Ant, 1963). At the western edge of its geographic range, /. isognomostoma occurs in the valley of the river “Ourthe” near La Roche-en-Ar- denne (Belgium) where it was recorded for the first time by Van Belle (1970). On the map provided by Kerney et al. (1983), the popula- tions in this region are shown as being iso- lated from the main region of distribution (Fig. 1). Irrespective of whether this is truly the case, the Belgian populations are peripheral and although /. isognomostoma might have established itself in this region a long time ago, it is equally possible that it represents a recent introduction. In any event, the species has been known in this region for at least 30 years. Generally, small and isolated populations are prone to effects of genetic drift and in- breeding, resulting in two possible, though not mutually exclusive, outcomes. Usually, ge- netic drift and inbreeding will lead to loss of al- leles and decreased heterozygosity, resulting in reduced evolutionary flexibility and a higher risk of extinction (Leberg, 1992; Frankham & Ralls, 1998; Saccheri et al., 1998). Alterna- tively, genetic drift and inbreeding may also lead to rapid genetic change, possibly en- hanced by an increased selection pressure near the edge of a species’ range (Levin, 1970; Soulé, 1973; Carson, 1975; Templeton, 1980). Hence, peripheral isolates may gener- ate biodiversity and may therefore be valu- able for conservation (Lesica & Allendorf, 1995). Furthermore, the Convention on Bio- logical Diversity explicitly recognizes that intraspecific variation needs protection be- "Royal Belgian Institute of Natural Sciences (RBINSc), Vautierstraat 29, B-1000 Brussels, Belgium “University of Antwerp (RUCA), Department of Biology, Groenenborgerlaan 171, B-2020 Antwerp, Belgium; vanriel @ ruca.ua.ac.be 2 VAN RIEL ET AL. 0.05) with mul- VAN RIEL ET AL. tyzO beSO €0S0 000`0- ZO000- Z21l0- 6200 0000 2320 98Ll'0 9820 ¿080 #0001 a (2500) (250'0) (50`0) (ELO'0) (S000) (pveo'o) (pzo0) (8100) (6r0'0) (Zr0'0) (570`0) (090`0) (ZE0'0) 7610 6800 0200 51:00 S000 6500 7500 8100 2800 S900 StO00 6800 8€00 AH (190°0) (r20'0) (8100) (ELO'0) (5000) (6E0'0) (ezoo) (810'0) (/50`0) (seoo) (250`0) (/10`0) (000'0) 6bL'O 2700 9500 800 S000 9900 EEOO 8100 1900 5500 ZEOO 1100 0000 НН (880) (eo) (680) (8£ 0) (ro) (зе) (p90) (sro) (9/0) (ezo) (oso) (070) (00`0) LUZ, ЗИ GEL три COLL SSL SELL ён у: 6SILL 9и би 00и а (гр (0 (201) (ol) (07) (eet) (eri) lez) (vez) (822) (89°) (51) (604) Егер 89° 256 BE OL Qt зи 19+ yok €szl szı $595: ses O£'OL id (Z'0) (Z'0) (z:0) (10) (z:0) (10) (10) (10) (20) (20) (10) (10) (10) 9! yl ri FE et el el Bl vl el Er al a у 902°0- 500`0- €Z0'0- ey 005`0 — > = = = = = 09820 2800 — — — 9 00/0 6860 000% 000% 000+ 000+ 000+ 000+ 05/0 EL6O O00} 0001 000'L g = LLO'O = = — = = = = = — — = У GP 9p ce 8 65 LL Or G ve ez Ss! 92 ee (N) W9d 6b0'0- 9/L0- 260`0 al 1520 0001 9260 — — z810 zero = = = — — — g €tZ'0 — r200 0001 0001 8180 8980 0001 000% O00! 000 0001 0001 v LE Or La 8 65 LL 8€ G ve ez Ss! 92 ze (N наэа ¿0001 /%0`0- ¿0001 .000'1 ay — 730`0 = = = = = = = = = = = 5 2280 9760 000'L 0001 000 000+ 0001 0001 000% 000'L 000+ 2970 9220 g 8210 _ = = = = = = = = — 8590 — #//`0 v Sp 9p La 8 65 LL 6€ G ve ez St 9% Le (N) HAgH ‚0980 ,9980 .SE8'0 6290 8820 9820 8650 000! 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The highest loadings were associated with shell measurements (DIA, HA and HS in the first root; HB and HA in the second root). Although all variables in the three roots con- tributed significantly (p < 0.05) to the separa- tion of the populations, generalizations that might appear from the canonical variables were not always significant for all compar- isons according to the Scheffé test. This sug- gested a more complex pattern of differentia- tion. However, the NMDS/MST graphs and UPGMA clustering based on the Mahalanobis distances (Fig. 3B) revealed, once again, that most variation was partitioned between re- gions (correlation with the matrix of genetic distances, r = 0.45; p < 0.01). DISCUSSION In contrast to chromosomal inversions, ho- mozygosity at allozyme loci is not directly cor- related with the position of a population near the periphery of a species’ range (Carson, 1955; Tigerstedt, 1973; Levin, 1978; Brus- sard, 1984), but is mostly attributed to the ef- fects of inbreeding and genetic drift in isolated populations (Shumaker & Babble, 1980; Coutellec-Vreto et al., 1994; Hafner & Shus- ter, 1996; Rankevich et al., 1996; Leijs et al., 1999). Hence, the relatively low levels of ge- For For Far 0.872* 0.035 0.867” 0.392* 0.333* 0.089* 0.21510: O 102 0.539* 0.078* 0.500* 0.761* 0.016 0.758* ONS 01377 01095 0.568* 0.152* 0.480” netic variation in most /. isognomostoma pop- ulations studied here might reflect the number or size of bottlenecks (founder events) that were involved in the colonization of new areas during inter- or postglacial periods (Hewitt, 1996; Leijs et al., 1999), so that subsequent genetic differentiation through isolation by dis- tance might have resulted in the current pop- ulation structure (Wright, 1978). Although this seems the most plausible scenario for neutral genetic markers, such as allozymes, the role of selection cannot be excluded (Ayala, 1999; Arter, 1990). High levels of allozyme differentiation, even between adjacent populations, is a common phenomenon in land snails, but mean Ест /. isognomostoma (0.550) exceeds the values found for most other outcrossing pulmonate land snails, such as Cepaea nemoralis (Ест = 0.308), С. hortensis (Ест = 0.206) (Ochman et al., 1987), Cristalabrum primum (Ест = 0.450) (Woodruff & Solem, 1990), Helix aspersa (Ест = 0.162) (Selander & Kaufman, 1975), Man- darina aureola (Fz, = 0.280), М. ponderosa (Рот = 0.130) (Chiba, 1993), and Solatopupa similis (Fgy = 0.568) (Boato, 1988). Assuming no methodological flaws (sam- pling error, interpretation of gels), the het- erozygote deficiency at HBDH in the periph- eral populations (AR1-2) and population AM is currently difficult to explain, particularly since the overall heterozygozity (H,) in population AM is exceptionally high compared to the other studied populations. The high H, in AM suggests that low levels of genetic variation are not a property of the species, but rather of local populations. Although we cannot ex- clude autogamy, heterozygous individuals at four other loci (PGM, DDH, PGD 8 EST-1) 10 VAN RIEL ET AL. and HW equilibrum in most populations, sug- gest that autogamy is not the general mode of reproduction. Multilocus heterozygosity levels are also within the range found in other out- crossing helicid snails (e.g., Nevo, 1978; Se- lander 4 Ochman, 1983; Falniowski et al., 1993). The incongruent results between direct (PAGE) and indirect (IEF) estimates of het- erozygosity are probably because IEF of EST provides both genotypic and non-genetic data, so that non-Mendelian variation will also contribute to variability (e.g., Jordaens et al., 1999, and references therein). Nevertheless, since the population groupings found with PAGE and DA were also found with IEF, the use of IEF profiles of EST may be useful for studying intraspecific variation. Despite the fact that the pattern of morphological differen- tiation is in good agreement with the elec- trophoretic data, this might also have resulted from adaptation to local climatic and/or eco- logical conditions or differences in age struc- ture. Since the Belgian populations are only slightly differentiated from neighbouring popu- lations and because they were only discov- ered in 1970, it is also possible that they have been introduced only recently. ACKNOWLEDGEMENTS PVR is funded by an IWT scholarship. This work was supported by the project “Genetic characterization of peripheral and isolated an- imal populations in Belgium, in execution of the Convention of Rio” (RBINSc) and FWO grant 2.0023.94. We thank H. Van Paesschen for his help with the illustrations. 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Cazzaniga Universidad Nacional del Sur, Departamento de Biologia, Bioquímica y Farmacia, San Juan 670, 8000 Bahia Blanca, Argentina; pmartin @ criba.edu.ar ABSTRACT The Argentinean apple snail Pomacea canaliculata is a recent invader in paddy fields in most Southeast Asian countries, where it has dispersed explosively since about 1980. Determinants of the natural range of P. canaliculata in its original area are poorly understood. In this study, fac- tors affecting its distribution in the southern limit of its native area in Buenos Aires Province were investigated. Salty, alkaline, poorly vegetated aquatic environments with high desiccation risk in the western part of the study area probably act as a natural barrier for its dispersal toward the south and west. Sites inhabited by P canaliculata are generally shallow, quiet, turbid, with low Na‘/(K* + Mg**) ratios compared to the uninhabited sites. Contrary to previous statements, P canaliculata is frequently found in streams, where it reaches the highest densities. Most of the inhabited environments are located on the northern slope of the Tandilia and Ventania moun- tains, this physical barrier, and not temperature, being probably the main factor impeding the species’ spread southwards in Buenos Aires Province. Pomacea canaliculata also inhabits iso- lated sites on the southern slopes of these mountains, where it appears to have been introduced. Its spread is slow within and among water bodies in this area. Repeated introductions, tropical climate, and integrated and flood-prone drainage systems probably account for the faster ex- pansion of this species in southeastern Asia. Key words: apple snail, native distribution, ecological factors, natural barriers. INTRODUCTION The Argentinean apple snail Pomacea canaliculata (Lamarck, 1822), is well known as a recent invader in paddy fields in most Southeast Asian countries (Halwart, 1994). The evidence of damage to aquatic crops (Yusa & Wada, 1999) and its ability to dis- place native species (Cowie, 1995) are of se- rious concern, especially in countries where rice is the staple food. The potential introduc- tion of apple snails to Australia prompted Baker (1998) to evaluate the extent of sus- ceptible areas in that country. The model used to determine the potential area that could be invaded was based mostly on published infor- mation on the climatic range of the species in South America. Baker (1998) predicted that other as-yet uninvaded Asian countries are potentially at risk of invasion. Nevertheless, the factors determining the natural range of Pomacea canaliculata in its native region are poorly understood. The 13 habitat of this species has usually been de- scribed only qualitatively, with no informa- tion on physico-chemical conditions. The spe- cies has been described as a lentic dweller, mostly inhabiting shallow lakes, ponds, and swamps (Hylton-Scott, 1958). It has also been recorded from prairie streams and vegetated backwaters (Cazzaniga, 1987). For more than a century it has been known that the southern limit of the species —the southernmost apple snail in the world —is somewhere in the south of Buenos Aires Province, Argentina (d’Orbigny, 1847; Hylton- Scott, 1958). Cazzaniga (1987) mentioned P. canaliculata from Calaveras stream (38° 04'S-59°18’W), this being the most southerly locality known until now. Knowledge of the factors affecting the species’ distribution at the southern edge of its natural range is of interest in understand- ing the factors determining its potential range in areas where it has been introduced re- cently. 14 MARTIN ET AL. MATERIALS & METHODS The sampling region extended from 36°S to 39°S in southern Buenos Aires Province. This range covers the southern part ofthe pampas (approximately 120,000 square kilometers) and includes the only two mountainous areas in the province (Ventania and Tandilia moun- tains). Rivers and streams have scant and very variable water flow, with a pluvial hydro- logical regime. The typical lentic habitats in the pampas are small, shallow lakes, many of them of temporary nature (Toresani et al., 1994). The climate of the region is temperate, with a marked humidity gradient from the NE (mean annual rainfall 900 mm) to the SW (600 mm). The whole region lies within the 14*C and 16°C annual mean isotherms. The sampling scheme included all the main drainage basins and many isolated streams and lakes. The limits of the sampling region were set by reference to published literature and personal experience: apple snails live al- most everywhere in Buenos Aires Province to the north of 36°S latitude (Hylton-Scott, 1958) and are naturally absent to the south of 38°S (Cazzaniga, 1981, 1987). They have never been found in La Pampa Province (i.e., west of 63°W), the westernmost records being two ponds located near 62°40’W (Estebenet & Cazzaniga, 1993). Seventy-six sites were visited in February and March 1998 (late summer) (Fig. 1). More complex drainage basins (i.e., with third- or fourth-order streams) were sampled at more than one location. The local reproductive season of P. canalic- ulata runs from late October to early April (Hylton-Scott, 1958; personal observations). During this period, the conspicuous, aerial, pink egg masses are an easily visible sign of the presence of apple snails, even if the snails are in low densities. Two people carefully inspected up to 200 m of the shores of every water body investi- gated, searching for egg masses on the emer- gent aquatic vegetation and other substrata. Living apple snails were searched for among the submerged vegetation, under stones, or buried in the substratum. Surrounding land was searched for empty shells. We consid- ered as inhabited by P. canaliculata only the sites where egg masses or live snails were found. Where possible, relative abundance of living apple snails was estimated by reference to search effort (number of snails >20 mm caught per hour). We recorded the number of other species of aquatic snails and the number of plant species (submerged, floating or emergent macrophytes) at every site. Also 16 environ- mental variables were recorded. Depth (m) and surface water velocity (m.s_') were mea- sured at different points within each site. Con- ductivity (mS.cm~') and pH were determined in situ with a multimeter (Horiba U-10). A sub- surface water sample for chemical analyses was collected at each site, immediately fixed with formaldehyde and stored in a refrigerator. Total and volatile suspended matter (9.Г') were determined according American Public Health Association (1981) methods. The con- centrations (теа.Г') of Na*, Ca**, Mg**, K* and SO, were measured with an inducted plasma emission spectrometer (Shimadzu ICPS 1000-III), and Cl, CO, and HCO,” were measured by titration. The dominant substratum was character- ized on an arbitrary scale: 1 (sapropel, mua), 2 (sand), and 3 (pebbles, boulders, lime- stone). The trophic resource availability was coded as 1 (low), 2 (medium), and 3 (high), through a visual estimation of the abundance of macrophytes, microphytes, riparian vege- tation and their debris. The environmental variables and the species richness of macrophytes and snails were standardized and examined by stepwise discriminant analysis (SDA) to find linear combinations of variables maximizing dis- tances between groups of sites. Group sizes were used to determine group membership probabilities. The variables were included one by one according to their individual discrimi- natory power and they were kept or removed after assessing if they improved or not the total discriminatory power. RESULTS Pomacea canaliculata was recorded at 25 sites. The absence of egg masses during the sampling period, which was within the repro- ductive season, proved to be a good indicator of the absence of the species; all habitats where apple snails were collected had egg masses, and in only two places where we found eggs were we unable to find live snails or empty shells. Six other species of snails were found in the 34°S ah Te PE Rd 3 > ' 15 ‚vr > ; G yu = Re 207 in A AN Se E j Е. - da, Vallimanca River_ > i ей > vA i 36°S ; Ie | : ; ' ! 385 ; ы ! ! | 1 EN Sy pe — 40°5 | i | a? Negro River - DISTRIBUTION OF POMACEA 15 UA Paraná River Se | o Colorado-River CS J Atlantic Ocean - O tp 2 NK FIG. 1. Map of Buenos Aires Province showing the 76 sites visited (inset map of Argentina). Squares and cir- cles represent lotic and lentic habitats, respectively. Empty symbols indicate sites surveyed but not inhabited by any species of snail. 16 MARTÍN ET AL. 16 14 - mwith snails (CC: 100%) Canonical correlation = 0.759 | 12 - without snails (CC: 64.7%) Wilks Lambda = 0.423°° | | 10 > 8 - о 5 = 03 Е Е 4- 2 4 0 + 2 3 = 4 = _ 45 35 25 15 05 15 25 35 45 discriminant score d—————— 0.000 m nn variables HCO, Mg” MSR Ca” "cor 0.631 0.187 0.509 0.363 FIG. 2. Frequency histograms of discriminant scores for sites not inhabited (n = 17) and inhabited (n = 59) by any species of snail (CC: percentage of correctly classified cases; r¿pp: correlation between discriminat- ing variables and canonical discriminant function; MSR: macrophyte species richness). area. The most widespread species (24 to 32 sites) were Chilina parchappii (d'Orbigny, 1835) (Chilinidae), Biomphalaria peregrina (d'Orbigny, 1835) (Planorbidae), and Heleo- bia parchappii (d’Orbigny, 1835) (Hydrobi- idae). Infrequent species (eight sites or less) were Gundlachia concentrica (d’Orbigny, 1835) (Ancylidae), Lymnaea viator (d’Or- bigny, 1835) (Lymnaeidae) and Physa sp. (Physidae; may be an introduced species). No snail species were recorded in 17 ofthe 76 sites (Fig. 1). A stepwise discriminant analysis was performed with the environmen- tal variables (snail species richness excluded) to characterize the sites with and without snails. High values of Ca** and macrophyte species richness, as opposed to high concen- trations of HCO, and Mg”, characterize the sites inhabited by at least one species of snail; eleven sites that were correctly classi- fied as not inhabited were considered not habitable (Fig. 2), and therefore excluded from further analyses (one stream and ten al- kaline lakes, nine of them located on the west end of the studied zone). Table 1 Summarizes mean and extreme val- ues of selected environmental variables in the sampled sites, discriminated as not habitable, potentially habitable for freshwater snails and, among the latter, those inhabited or not by P. canaliculata. To characterize the habitats with and with- out apple snails (n = 65; Fig. 3), we performed a stepwise discriminant analysis with the vari- ables used in the former analysis plus snail species richness. The variables Na* and water velocity, as opposed to K*, Mg”, total suspended matter and depth, discriminate be- tween sites not inhabited and inhabited by Р canaliculata (Fig. 4). They mostly inhabit non- shallow sites, with null or low water velocity, high content of suspended material, and low Na*/(K* + Mg**) ratios (Table 1). Abundance was higher in streams (92.1 snails.h*) than in lakes (41.3 snails.h?) (t test for unequal variances: t = 2.78, df = 13.18, p < 0.02). The highest densities in the streams were found in mats of Ludwigia sp. (Onagraceae), where high numbers of apple snails were immersed in flocculent sediments DISTRIBUTION OF POMACEA 17 TABLE 1. Mean and extreme values of selected environmental variables (conductivity values are given in mS.cm !, ionic concentrations in тед.Г', depth in m, water velocity in m.s* and total suspended matter in g.1?). Not habitable All sites for snails n 76 11 pH 8.35 9.03 7.30-9.90 8.48-9.40 Conductivity 3.50 16.37 0.15-83.50 2.10-83.50 Na* 30.79 160.29 0.47-820.00 11.40-820.00 Ca** 1.39 0.74 0.36-3.69 0.36-1.47 Mg** 1151] 2.14 0.33-4.84 0.61-4.84 K* 0.42 157. 0.04-6.66 0.24-6.66 cr 14.59 79.62 0.44-389.60 2.80-389.60 $047 6.76 30.88 0.04-180.00 1.74-180.00 СОЗ” 1.29 7.04 0.00-21.90 0.53-21.90 HCOS3” 8.10 19.43 1.10-33.80 7.02-33.80 Na*/(K* + Mg”) 10.03 36.23 0.53-71.30 4.87-71.30 K*/Na* 0.040 0.014 0.007-0.143 0.008-0.036 Depth 0.83 0.70 0.15-3.00 0.35-2.00 Water Velocity 0.17 0.02 0.00-0.77 0.00-0.18 Mg**/Ca** 1.28 2.95 0.43-7.81 0.76-7.81 Total Suspended Matter 0.023 0.043 0.000-0.099 0.015-0.099 Habitable Not inhabited by Inhabited by for snails Р canaliculata P. canaliculata 65 40 25 8.24 8.17 8.35 7.30-9.90 7.30-9.54 7.30-9.90 1.32 1.48 1.06 0.15-3.33 0.15-3.33 0.23-2.89 8.87 10.34 6.53 0.47-28.61 0.47-28.61 0.85-19.84 1.49 1.44 1.59 0.38-3.69 0.45-2.94 0.38-3.69 1.40 Wee 1.54 0.33-2.81 0.33-2.70 0.56-2.81 0.22 0.20 0.25 0.04-0.60 0.04-0.35 0.07 -0.60 3.59 4.07 2.81 0.44-11.44 0.44-11.44 0.52-9.96 2.68 3.59 1623 0.04 -17.34 0.04-17.34 0.07-15.31 0.31 0.37 0.22 0.00-2.12 0.00-2.12 0.00-1.43 6.18 6.36 5.88 1.10-11.40 1.61-11.40 1.10-9.95 5.60 6.73 3.78 0.53-16.76 0.59-16.76 0.53-10.12 0.044 0.036 0.057 0.007-0.143 0.007-0.113 0.014-0.143 0.85 0.783 1.04 0.15-3.00 0.15-3.00 0.30-1.70 0.20 0.22 0.15 0.00-0.77 0.00-0.77 0.00-0.67 1.00 0.93 1.10 0.43-3.06 0.43-1.43 0.52-3.06 0.019 0.013 0.029 0.000-0.088 0.000-0.062 0.000-0.088 rich in organic matter. The midstream surface velocity ofthe sampled streams was generally low (0.22 m.s* + 0.03, mean + SE), but oc- casionally apple snails were found adhered to different substrata in faster running water. The frequency of water bodies inhabited by P. canaliculata on both sides of the Tandilia and Ventania mountains was clearly different (Fig. 3). Of the 31 habitable sites located to the north of the mountainous area, 19 (61%) were inhabited by apple snails (83% of the lentic habitats; 56% of the lotic ones). To the south of the mountains, only 18% of the sites had P. canaliculata: it was found in only six, mostly isolated locations in four different basins (from west to east: Chasicö, Sauce Grande, Quequén Grande and El Durazno). To test if differences between inhabited and not inhabited sites could be explained by their location on the northern or southern slope we performed a SDA using only the six variables distinguishing inhabited and not inhabited sites. The habitats located to the north of the mountains differ significantly from the south- ern sites by their higher K*/Na* ratios (Fig. 5). Within the northern slope, high values of depth and Mg” characterize the habitats with apple snails (SDA with the 18 environmental variables, Fig. 6). Eight of the 11 lotic environ- ments with no P. canaliculata on this slope are located near the stream sources (Fig. 3); five of them are the only streams running through the plains between Ventania and Tandilia mountains and have the lowest concentra- tions of Mg**. It was not possible to discrimi- nate between sites inhabited and not inhab- 18 MARTÍN ET AL. Vallimanca de sé ci gees and : р 7 _> € { Atlantic Ocean FIG. 3. Map of the study area showing the 65 sites considered habitable for snails. Squares and circles lo- cate lotic and lentic habitats respectively. Empty and filled symbols indicate sites not inhabited and inhabited by Pomacea canaliculata respectively. ited by P. canaliculata on the southern slope: no individual variable showed a significant dif- ference (all Wilk's Lambda > 0.89, p > 0.05). DISCUSSION Ecosystems with alkaline waters, high Mg'**/Ca”* ratios and poor in macrophytes are adverse for snails within the study area. Low concentrations of Ca** and high concentra- tions of Mg** (either absolute or relative to Ca”) are a constraint for the distribution of freshwater snails (Lodge et al., 1987; Mad- sen, 1987; Savage & Gazey, 1987; Brown, 1994). A positive relation is found between snail and macrophyte species richness, al- though this correlation may in part be be- cause macrophyte distribution is also affected by the water chemistry (Okland, 1979; Pip, 1987). The predominance of salty, alkaline, poorly vegetated aquatic environments with high desiccation risk in the western part of Buenos Aires and eastern part of La Pampa provinces (Damario & Pascale, 1993; Gilbert et al., 1996) is probably a natural barrier for the dis- persal of snails and other freshwater organ- isms toward the south and west. This barrier is not evident from topographic or hydro- graphic standpoints and was not appealed to in Argentinean studies on biogeography (e.g., Ringuelet, 1975; Bonetto, 1973). Irrespective of their geographical position relative to the mountains, sites inhabited by P. canaliculata generally are shallow, quiet and turbid, with low Na*/(K* + Mg**) ratios as com- pared to the uninhabited ones. Most of the in- habited environments are located on the northern slope of the mountains and these dif- fer from the southern ones in their low K*/Na* ratios. It is probable, thereby, that the geo- graphic position biased the selection of the distinctive variables of the habitats inhabited by P. canaliculata, especially in the chemical aspect. DISTRIBUTION OF POMACEA 19 105 8 - with P. canaliculata (CC: 84.0%) Canonical correlation = 0.711 | , = Lis: 6 | | without P. canaliculata (СС: 87.5%) | Wilks' Lambda = 0.494 | L 4 - г 2. о 5 u O +— $ Е 23 4 =] 6 - 8 - 10 - -45 3.5 -25 -1.5 0.5 1.5 25 3.5 45 discriminant score variables Na° VEL Mg” D К TSM ГСО -0.283 -0.146 0.174 0.252 0.254 0.407 FIG. 4. Frequency histograms of discriminant scores for habitable sites actually inhabited (п = 25) and not inhabited (п = 40) by Pomacea canaliculata (CC: percentage of correctly classified cases; r,..-: correlation between discriminating variables and canonical discriminant function; VEL: water velocity; D: depth; TSM: total suspended matter). Shallowness is probably related to the risk of becoming temporally dry, and also to a lower stability. Water bodies in the area are al- most exclusively fed by rainfall, summer being the season with lower water level. Droughts are frequent in this area (Damario & Pascale, 1993), and many water bodies dry up (Gron- dona, 1975; Toresani et al., 1994). Though adult Р canaliculata are able to survive for several months out of water (d’Orbigny, 1847), summer dryness might lead to local extinctions as a result of a combination of high temperatures, dehydration, hypoxia, starva- tion or predation. The resistance of juveniles to these factors may be lower than that of adults, as in Pomacea paludosa (Say, 1829) (Turner, 1996). In this region it takes approximately 17 days from oviposition to hatching (unpublished data). Even if a water body does not dry up completely, recruitment may be affected by the withdrawing of the water edge (especially in streams), which may lead newly hatched snails to drop on drying soil. Other Pomacea species attain their highest densities in more permanent habitats (Kushlan, 1975; Donnay & Beissinger, 1993). Some literature states that P. canaliculata inhabits lentic water bodies, while Pomacea insularum (d’Orbigny, 1835) lives in lotic habi- tats only (Hylton-Scott, 1958; Bachmann, 1960). Our data show that P. canaliculata also occurs frequently in streams, where it reaches its highest densities, though quantitatively confirm that it prefers still or slow-running wa- ters. Water speed does not seem, however, to be a limiting factor per se in most uninhabited streams, since on the one hand midstream surface velocities are generally low and on the other the species has been observed to endure high velocities. Probably the influence of water velocity is related to the general in- cline of the stream section, which in turn af- fects its hydrological regime. The preference for sites with high content of suspended matter probably is not supported either by trophic needs or by the nature of the sediments, since suspended organic matter and sediment types were not selected by the stepwise discriminant analysis. Maybe it is more related to the protecting turbidity that re- duces the probability of predation. The influence of Mg*™ in the northern zone is probably related to extremely low concen- 20 MARTÍN ЕТ AL. 16 № northern slape (CC: 80.6%) E = = A | rrelation = 0573 12 - southern slope (CC: 76.5%) ee ee 8 a S44 e Ф = 4 o м | E || ==] | ES lias =} 4 - L— 8 - | 12 45 -3.5 -25 -1.5 0.5 155 25 3.5 45 discriminant score С variables K Na’ Tcor 0.304 0.601 FIG. 5. Frequency histograms of discriminant scores for habitable sites in the northern (n = 31) and south- ern slope (п = 34) of the Ventania and Tandilia mountains (CC: percentage of correctly classified cases; гор. correlation between discriminating variables and canonical discriminant function). trations in a particular group of short intermit- tent rivulets located in the inter-mountain plain, a flat area with scarce and scattered water bodies where apple snails are absent. Pomacea canaliculata is able to survive and reproduce in colder regions, such as Rio Colorado Valley (Fig. 3; Cazzaniga, 1981). The minor temperature gradient between the northern and southern boundaries of the study area (2°C) suggests that temperature is not the factor limiting the distribution of apple snails in the south of Buenos Aires Province. Even if aridity limits the distribution of apple snails towards the west, probably it is not a limiting factor towards the south, since cli- matic conditions to the south of Tandilia are not dryer than those found on the northern slope of Ventania. The discontinuity between the northern and southern drainages is the physical barrier lim- iting the spread of P. canaliculata, in spite of our records in six localities (four isolated basins) to the south of it. Sport fishing is in- tense in three of these four basins, fishermen coming frequently from northern locations (Grossman, 1993; personal observations), and they may have introduced P. canaliculata into these areas. The use of apple snails as fishing bait has been one of the reasons for its rapid dispersal in southeastern Asia (Ander- son, 1993; Wada, 1997). Fish sowing activities may also have caused accidental introduction of P. canaliculata. At present, the southern- most location is Paso de las Piedras reservoir, Sauce Grande basin (38°24'S), where the apple snails were first seen in 1991 (Kaspar Delhey, pers. comm., 1991; this study). The distribution of Р canaliculata is essen- tially tropical and subtropical, including the La Plata basin (Ihering, 1919). Maybe its southerly distribution was reduced to a mini- mum during the last glacial age (Pleistocene- Holocene boundary), when the cold, arid cli- matic conditions of present-day Patagonia extended up to 750 km northward, north of the Paraná river delta (Fig. 1; Iriondo, 1994). Sub- tropical conditions advanced to southern Buenos Aires Province during the middle Holocene (8-6 Kyr BP), extending the maxi- mum potential distribution of P. canaliculata to the south. There are numerous Holocene fos- sil deposits containing freshwater snails on the southern slope of Tandilia and Ventania mountains (e.g., Frenguelli, 1945; Kerlleñe- DISTRIBUTION OF POMACEA 21 S 7 4 | with P. canaliculata (CC: 84.2%) || without P. canaliculata (CC: 66.7%) | au frequency o Te Set | Canonical correlation = 0.711 2- Pz Wilks' Lambda = 0.494*** | 4- (ae Si 5 | -4.5 -3.5 -25 -15 05 15 2:5 35 45 discriminant score variables Mg” D 0.388 0.712 ГСОЕ FIG. 6. Frequency histograms of discriminant scores for habitable sites in the northern slope of the Ventania and Tandilia mountains actually inhabited (n = 19) and not inhabited (n = 12) by Pomacea canaliculata (CC: percentage of correctly classified cases; r.p-: correlation between discriminating variables and canonical dis- criminant function; D: depth). vich, 1989; Bonadonna et al., 1995). Though they yield the same snail species that now occur in the area, none of them contains P canaliculata, although P. canaliculata is com- mon as a fossil on the northern slope (Frenguelli, 1945; Camacho, 1966; Dangavs, 1988). The only fossil record from the south- ern slope corresponds to shell fragments from the late Pleistocene near El Durazno stream that Frenguelli (1921) identified as P. canalic- ulata. However, the species was not recorded among the freshwater snails found in more re- cent sediments there. Pomacea canaliculata seems to have been unable to surmount the obstacle of the Tandilia and Ventania mountains in the past. However, these mountains were not a barrier for the other freshwater snails living in the area, which are present all over Buenos Aires Province and northern Patagonia (Castel- lanos & Landoni, 1996). As compared to these snails, Р canaliculata has a larger body size and takes longer to reach maturity, and it is dioecious, with aerial eggs. All these char- acteristics could theoretically lead to lesser dispersal ability. Pomacea canaliculata appears unable to spread even within one same basin in the southern slope. In the Chasicó basin, it was collected only from one lake (Laguna Los Chilenos) and the proximal sections of two streams flowing in and out of it, although it is known to have been present in that lake since 1985, when we recorded high densities of both old shells and living snails. In Sauce Grande basin (first record 1991) and Que- quén Grande basin (first record 1983; Caz- zaniga, 1987), they were found only in one lentic site each. The most effective natural dispersal mechanism seems to be floating downstream, but it does not appear to be enough to generate populations downstream of the introduction point even after eight years or more. The slow dispersal of P. canaliculata within and among water bodies on the southern slope of the mountains strongly contrasts with its fast expansion in southeastern Asia. Sev- eral factors may contribute to an explanation of these differences. The most obvious one is the initial interest in actively spreading a po- tential economic resource before realizing the risks. Apple snail stocks were introduced sev- eral times into almost all countries in that re- 22 MARTÍN ET AL. gion (Halwart, 1994; Naylor, 1996; Cowie, in press). Pomacea spp. attain maturity at an early age and reproduce all year round in tropical and subtropical climates (Milward de Andrade et al., 1978; Pointier et al., 1988; Donnay 8 Beissinger, 1993; Halwart, 1994), two factors that probably enhanced the quick establishment and spread of apple snails in southeastern Asia (Naylor, 1996; Cowie, in press). Periodic floods caused by summer monsoons may have contributed to its fast spread among and within Asian water bodies (Anderson, 1993; Wada, 1997). In Buenos Aires Province, growth of P canaliculata is slower than in tropical areas, age at maturity is greater, and thermal sea- sonality determines a short reproductive pe- riod (about six months), and a winter resting period (Bachmann, 1960; Estebenet & Caz- zaniga, 1992, 1993, 1998). In addition, on the southern slope of the Buenos Aires mioun- tains, rivers are relatively short and run paral- lel to each other and into the Atlantic Ocean, with almost no connection among their basins, these conditions precluding a more ef- fective expansion. ACKNOWLEDGMENTS This work was funded with grants by CON- ICET (“Consejo Nacional de Investigaciones Cientificas y Técnicas”) and “Agencia Na- cional de Promoción Científica y Técnica”, Ar- gentina. ALE is a researcher in CONICET and NJC is a researcher in “Comisión de Investi- gaciones Científicas de la Provincia de Buenos Aires”. 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PIP, E., 1987, Species richness of freshwater gas- tropod communities in central North America. Journal of Molluscan Studies, 53: 163-170. POINTIER, J. P., A. THERON & D. IMBERT-ES- TABLET, 1988, Decline of a sylvatic focus of Schistosoma mansoni in Guadeloupe (French West Indies) following competitive displacement of the snail host Biomphalaria glabrata by Am- pullaria glauca. Oecologia, 75: 38-43. RINGUELET, В. A., 1975, Zoogeografia y ecología de los peces de aguas continentales de la Ar- gentina y consideraciones sobre las áreas icti- ológicas de América del Sur. Ecosur, 2: 1-122. SAVAGE, A. A. 8 G. M. GAZEY, 1987, Relation- ships of physical and chemical conditions to species diversity and density of gastropods in English lakes. Biological Conservation, 42: 95-113. ] : TORESANI, N. I., Н. L. LOPEZ & 5. E. GOMEZ, 1994, Lagunas de la Provincia de Buenos Aires. Ministerio de la Producción de la provincia de Buenos Aires Ed., 108 pp., La Plata. TURNER, R. L., 1996, Use of stems of emergent plants for oviposition by the Florida applesnail, Pomacea paludosa, and implications for marsh management. Florida Scientist, 59: 34-49. WADA, T., 1997, Introduction of the apple snail Po- macea canaliculata and its impact on rice agri- culture. Proceedings of the International Work- shop on Biological Invasions of Ecosystems by Pests and Beneficial Organisms, pp. 170-180, Tsukuba. YUSA, Y. & T. WADA, 1999, Impact of the introduc- tion of apple snails and their control in Japan. Naga, The ICLARM Quarterly, 22: 9-13. Revised ms. accepted 25 June 2000 MALACOLOGIA, 2001, 43(1-2): 25-32 LIFE CYCLE AND POPULATION DENSITY OF THE PEST SLUG ARION LUSITANICUS MABILLE (MOLLUSCA: PULMONATA) ON GRASSLAND Brigitte Grimm Institut für Zoologie, Universitátsplatz 2, A-8010 Graz, Austria; grimm O kfunigraz.ac.at ABSTRACT Arion lusitanicus is a univoltine, semelparous slug found in Austria. Hatchlings were found both during the autumn, in which the eggs were laid, and in the following spring. By far the largest part of this generation matured the following summer (June to August), with peaks of egg-deposition noted in late August and early September. Population density was measued by MRR (mark-re- lease-recapture) and hand-searching; the results obtained by the two methods did not differ sig- nificantly. In June 1996 (the peak month for 1996), MRR gave a density of 1.6 slugs пт on grass- land. The high recapture rates of individually marked slugs, as well as the fact that their life parameters fell well within the range of unmarked individuals, is a proof of their unaltered vitality and thus of the applicability of the new method in the field. The effects of different weather con- ditions on the main life-parameters are discussed. Key words: Arion lusitanicus, life cycle, density, transponders, individual marking, weather conditions. INTRODUCTION Like other arionid slugs (Beyer & Saari, 1978; South, 1989), A. lusitanicus Mabille, 1868, is known to have a univoltine and semelparous cycle in western and northern Europe (Quick, 1952; Davies, 1987; Cheval- lier, 1971; Risch 4 Backeljau, 1989; von Proschwitz, pers. comm.). From the scattered evidence of life-history data presently avail- able, A. lusitanicus was assumed also to be annual and semelparous in central Europe (Graber & Suter, 1987; Grimm, 1991, 1996). The present investigation was designed to elucidate the details of the life cycle of this economically highly important pest slug (e.g., Schmid, 1970; Frank, 1998; von Proschwitz, 1997; Fischer & Reischútz, 1998), because the main life-history parameters as well as the knowledge of population densities are of basic relevance for determining the timing of efficient control methods. А new method of individually marking slugs by injecting magnetic transponders, which has yielded very promising results in the lab- oratory (Grimm, 1996), was applied under field conditions for the first time. А comparison of life cycle data of marked and unmarked Slugs at the same site, would allow a definite conclusion about the applicability of the method in the field. 25 The importance of different weather condi- tions as factors determining slug development (e.g., Dainton, 1954; Crawford-Sidebotham, 1972; Young et al., 1993; Keller et al., 1999), was also investigated in the present study. MATERIALS & METHODS Study Area The study was conducted on grassland, which was mown twice a year. It was situated near St. Marein/Graz in Austria (47°1'N, 15° 41'E), elevation 300 m). The study area con- sisted of a small strip (10 to 12 m wide) run- ning from north to south for 200 m between a conventionally managed field to the east (used only in 1995), and stream-side vegeta- tion to the west and south (used in 1995 and 1996). No herbicides and pesticides were used. A total area of 290 m? was used in 1995, and 260 m? in 1996. In June 1996 the meadow had 90-95% plant cover. The sam- pling periods ran from May to November 1995 and from March to December 1996. Other slug species (Kerney et al., 1983) found on the site were: Arion subfuscus (Dra- parnaud, 1805); Arion fasciatus (Nilsson, 1822); Limax maximus Linnaeus, 1758; Mala- colimax tenellus Muller, 1774; Deroceras 26 GRIMM reticulatum (Muller, 1774); and Deroceras laeve (Muller, 1774). Their densities were not determined. Weather Conditions Monthly arithmetical means of temperature and precipitation are compared with a 30-year mean (1961-1990). Temperature was ob- tained from the Metereological Station at Graz-Thalerhof (distance to the site: 19.2 km) and precipitation from the Metereological Sta- tion at Gleisdorf (distance to the site: 8.8 km). Parameters from two different metereological stations were used, because their data were expected to resemble the situation of the ac- tual site best (Lazar, Institut fur Meteorologie, Universitat Graz, 1999, pers. comm.). Temperature: Greater monthly fluctuations of temperature were observed in 1996 (S.D. + 8.66) than in 1995 (S.D. + 8.06), the latter also being the warmer year (mean 9.77°C in 1995, 8.83°C in 1996). The winter of 1995/96 was significantly colder than average, and also than 1994/95 (mean 95/96 1.5° (S.D. + 4.5); 30-year mean 2.1 (S.D. + 4.6); mean 94/95 5.1 (S.D. + 6.9). Spring (February— May) was warmer in 1995 (mean 8.6°, S.D. + 4.6) than 1996 (mean 5.6°, S.D. + 7.4) as was the late summer/autumn (July-October) of 1995 (mean 16.65”, S.D. + 3.95) com- pared to the same period in 1996 (mean 15.287, S:D: = 348). Precipitation: 1996 proved to be significantly wetter than 1995 (p = 4.8, d.f. = 22; 1995: mean = 64.87, S.D. + 39.45; 1996: mean = 76.83, S.D. + 39.03), except for an extremely dry July (54.4 mm below 30-year’s average). The late spring and early summer (April- June) of 1995 was dry (55.9 mm below 30- year’s average) compared to the same period of 1996 (54.3 mm more than 30-year’s aver- age). 1995 also ended in a drier autumn than 1996 (September-November; 1995: 115.6 mm; 1996: 270.2 mm). The winter 1995/96 (December-April) had high precipitation (33.1 mm above average) and thus snow-coverage for three months. Refuge Traps To assess the local density of slugs, 50 x 50 cm squares of 3 mm hardboard covered with 5 cm polystyrene were used as shelter traps. In 1995, 54 traps were placed 2 m apart in a rectangular grid over the entire study site. Four rows of ten traps each were laid on the grassland area, another row of ten traps on the oil-pumpkin field, and a row of only four traps was placed in the stream-side vegeta- tion (stream meandering prevented the set- ting of further traps). In 1996 only the central four rows (40 traps) on the grassland were used. This was due to increasingly heavy un- dergrowth in the brook-vegetation area, and because permission to place traps on the con- ventionally managed field could not be ob- tained for the second year. The traps were placed on short-mown vegetation patches of identical size two weeks before the start of the investigation in 1995 and remained in place for both seasons; damaged traps were re- placed immediately. The living vegetation be- neath the traps disappeared within two weeks of their placement; dead plant remains could be found for a further four weeks. Marking A method of individually marking A. lusita- nicus by magnetic transponders (Grimm, 1996) was used for the first time in the field; 92 slugs were marked in 1995, and 139 slugs in 1996. Between July 12 to 18 1995, all 57 A. lusitanicus found under the shelter traps were given a unique mark directly in the field and replaced exactly where they had been found. In 1996, two similar marking events (one in June, one in August) were carried out. Recording of Activities The site was searched twice a day each day, throughout the periods May to November 1995 and March to December 1996. All A. lusitanicus found under the shelter traps and the area in between were recorded and ex- amined for the presence of a transponder mark using the hand-reading instrument (Grimm, 1996). All mating pairs over the site, and all egg-batches found under the shelter traps, were also recorded. Determination of Developmental Status of A. lusitanicus Every week 20 A. lusitanicus, representing the full size-range at a time, were collected from a nearby field (distance 100 m) and taken to the laboratory for identification (lower genitalia; Kerney et al., 1983; Quick, 1960) LIFE CYCLE 8 POPULATION DENSITY OF A. LUSITANICUS 27 TABLE 1. Population density in 1995 and 1996 as determined by hand-searching (means, S.E., sig- nificance in difference between 1995 and 1996 (U-test; a = D. 05; ** highly significant, * significant, n.s. not significant) over the whole site (290 m? in 1995, 260 m? in 1996); shelter traps and between traps were combined (juveniles included in autumn). 1995 mean SIE? March = = April SE = May 52.2 4.02 June 136.8 12.8 July 60.9 7.4 August 39.8 5.9 September 33:3 3.4 October ИГ 158) November 122 OS December = and for determination of their developmental status. When sperm could be found in the her- maphrodite duct , slugs were considered to be “male-mature”; before that they were regis- tered as “juvenile”. Slugs showing an en- larged albumen gland and eggs in their oviduct were considered “female-mature”. These results were complemented by field- observations of mating and egg-laying. Statistical Analysis Population densities were estimated ac- cording to mark-release-recapture (MRR) methods of Jolly in 1995, and Jackson’s Pos- itive Method in 1996 (Begon, 1979). Descrip- tive statistics, t-test and Mann-Whitney U-test were calculated using SPSSO. RESULTS Seasonal Level and Absolute Densities In both years, a peak mean density occured in June (41.36% higher in 1996 than 1995) and was followed by a continuous decrease for the rest of the season (Fig. 1). The mean number of slugs for both years differed highly significantly from May to August, and by Sep- tember recorded numbers were still 57.8% above those reached in 1995 (significant) (Table 1). Specimens were marked in both years, al- lowing the population densities to be esti- mated according to the mark-release-recap- 1996 es Significance between mean Sees 1995 and 1996 23.4 13 = 39.8 1.8 = 109.3 7.9 ia 228.9 18 di 153.4 19.5 E 104.2 18% de 49.4 6.2 6 8.06 2.9 n.s 1.04 0.4 n.s 0.34 0.2 == ture (MRR) methods. Over the entire site, in- cluding shelter traps, on July 14, 1995, the density estimated by MRR was 0.22 m? whereas the arithmetical mean of the num- bers counted by hand-searching for that month was 0.21 пт? (Fig. 1), which was only 4.5% less than the number estimated by MRR (difference not significant). In 1996, the esti- mate for June 18 was 1.6 m, compared to the number counted by hand- -searching for the same day which was 1.4 m ?. The differ- ence between the two months was again not significant (12.5%). A highly significant increase in the number of egg-batches per trap from 1.15 in 1995 to 2.46 in 1996 was noted. Life Cycle In the present study, male-mature individu- als were first observed at the beginning of Au- gust (Table 2). The first spermatophores were found in the reproductive system one week before in 1995, and two weeks before in 1996. Part of the population remained in this state until October, while the remaining individuals entered female maturity (egg-laying phase) soon after (1995) or even before (1996) the first copulations were observed in the field (Table 2). According to data obtained from anatomical dissections, the first female-ma- ture slugs were found in the last week of July 1995, and in the last week of June 1996. The peaks of egg-deposition were in early Sep- tember 1995 and late August 1996 (Fig. 2). The period of egg-laying was longer in 1996 28 GRIMM mean numbers of slugs /m? а —— №1995 (11996 — 08: == ~ =. = 0,7 +— =—— une = 06 — — ay 0,5 ~——----— 0,4 4 A 0,3 pare 0,1 - Hi as al l March April June July Aug Sept Oct Nov Dec FIG. 1. Seasonal level of activity of A. lusitanicus in 1995 and 1996 (mean number of slugs per m? and per search). Data obtained by hand-searching over the whole area; shelter traps and between traps were com- bined (juveniles not included in autumn). TABLE 2. Reproductive data of marked and unmarked specimens according to field observations in 1995 and 1996. 1995 1996 Unmarked Marked Unmarked Marked Dates Copulations Aug. 6-Oct. 2 Aug. 6-Sep. 19 Aug. 7-Sep. 21 Aug. 11-Aug. 27 Dates Egg-laying Aug. 25-Sep. 29 Aug. 25-Sep. 19 Jun. 7-Nov. 11 Jun 7-Sep. 1 Numbers Egg-batches 0.87 (+ 1.06) >= 2.46 (+ 2.17) ~ (mean, + S.D.) per trap Dates Hatching Oct. 2-Oct. 29 = Sep. 17-Nov. 14 = than in 1995, extending to the middle of No- DISCUSSION vember (Table 2). Hatching was first observed in mid-Sep- Methodology tember 1996, 15 days earlier than in 1995 (Table 2). Since Oekland (1929), shelter traps have commonly been used for determining slug abundance. The type of trap used here, and Marked Specimens previously by Kaiser et al. (1993) and Grimm et al. (1997), represents an improvement for The restricted life-history data obtained this sort of field trial, even in comparison with from marked specimens was within the range pure cardboard (Grimm, 1991; Young et al., of data deriving from unmarked slugs (Table 1996; Hawkins et al. 1998), because it pro- 2). Recapture rates for marked slugs were vides rather more stable microclimatic condi- 83.2% in 1995, and 76.7% in 1996. tions (Grimm & Calame, in prep.), and addi- LIFE CYCLE 8 POPULATION DENSITY OF A. LUSITANICUS 29 1531 1-15 1530 1-15 1531 1-15 Sep Oct Nov FIG. 2. Seasonal frequency of egg-deposition (in %) of A. lusitanicus in 1995 and 1996. tional attractants, like baits, need not be used. Moreover, the traps can stay in the field for a full season or longer. Good microclimatic con- ditions are also provided by the shelter traps used by Hommay et al. (1991), and a direct comparison between these two in a field trial would provide valuable information for com- parison of data collected in the different in- vestigations (Grimm & Calame, in prep.). It would be very desirable to try to unify shelter- trap methodology generally. Differences in attractiveness of the traps to different stages of slug, as shown by the much higher density in June compared with earlier in the same year, can either be explained by differences in the behaviour of the stages themselves (Grimm, in prep.), or by changes in the microclimatic conditions under the traps during the year. The life-long duration of the individual marking technique, coupled with the easy and reliable readability of the codes, as known from laboratory experiments (Grimm, 1996), were also important advantages in the pres- ent investigation, as were the high recapture rates of marked individuals. The fact that marking did not alter the animals' vitality under natural conditions was also demon- strated for the first time in this field trial, as the mating and egg-laying periods of the marked slugs fell well within the range of untreated specimens. Life Cycle Like other species of arionid slug (Cheval- lier, 1971; Beyer 8 Saari, 1978; South, 1989), A. lusitanicus has an annual, semelparous cycle in most parts of Europe (Quick, 1952; Davies, 1987; Chevallier, 1971; von Pro- schwitz, pers. comm.), showing male maturity in summer and egg laying in autumn, whereas in southwestern Europe this species mates in January-February (Castillejo, 1997), and egg- batches occur in late autumn and winter (Castillejo, pers. comm.). However, this incon- sistency in the biological data between south- west and all other parts of Europe could also be the result of taxonomic confusion, a possi- bility well worth investigating. In A. /usitanicus, as in A. intermedius (Normand, 1852), egg- batches and young hatchlings were found in autumn and spring (South, 1989). Thus, over- wintering can take place at both those stages. The hatching date is mainly dependent on in- tegrated temperature (Kaiser et al., 1993), as was first demonstrated for insects (codling moth) by Shelford (1927). At any one time, body size can differ re- markably among individuals of a single gen- 30 GRIMM eration, with distinctly larger specimens being found in early spring (Grimm, 1991), and which most probably mature by early summer (Chevallier, 1971). This is also suggested by the timing of the first egg-batches, which were found at the beginning of June, the next only occuring in August. However, for the majority of the generation, it is probably not body size that is the main cause of maturity, as sug- gested by Lusis (1961), but rather synchro- nization with season (Smith, 1966), that is, day length (Sokolove 8 McCrone, 1978) and temperature (Rollo, 1983). These factors re- sult in a period of mating and egg-laying last- ing approximately two months for A. /usitani- cus, which is independent of the body size or weight of the slugs. Records of female-mature slugs earlier in the year than male-mature ones might result from the ability of A. /usita- nicus to self-fertilize (Foltz et al., 1982; von Proschwitz, pers. comm.). This and the ap- pearance of slightly acyclic specimens may provide security for this generally univoltine species. Population Densities Like A. subfuscus, A. lusitanicus reaches its peak population density in the early summer (June), whereas two other arionid slug species (A. fasciatus and the A. hortensis agg regate of species) show a distinct shift in their maximum densities towards the autumn (Barnes & Weil, 1944). This suggests that A. subfuscus and A. lusitanicus are closely re- lated not just morphologically (Backeljau, pers. comm.), but also ecologically. A heavy decline in numbers in the dry months of the summer (July onwards) and also in winter could be seen in the present investigation, agreeing with the findings of Runham & Hunter (1970) for A. hortensis. One of the causes of the increased pop- ulation density in the second year of the investigation may be the _ self-reinforcing mechanisms known to operate in growing populations (e.g., Wolda & Kreulen, 1973). The number of egg-batches was observed to be strongly linked to population density, as the significantly higher numbers of reproductive slugs in 1996 (64.4%) also produced signifi- cantly higher numbers of egg batches per trap (33.3%). Moreover, the prevailing weather conditions (e.g., Webley, 1964; Newell, 1968; Crawford-Sidebotham, 1972; Dainton, 1954; Young et. al., 1993), as well as the traps used as additional weather-protected egg-laying and resting sites, may also have contributed to the high population densities in 1996. How- ever, population densities in the present in- vestigation do not seem artificially high, be- cause in similar locations densities up to 10 m have been found in Sweden (von Proschwitz, 1994), up to 57.2 m” in Switzer- land (Frank, 1998), and up to 70 m? in Aus- tria (Grimm, unpublished). Unfavourable climatic conditions are gener- ally known to have a negative, delaying effect on feeding, growth and reproduction of arionid slugs (Laviolette, 1950; Hunter, 1968; Cheval- lier, 1971; Burenkov, 1977; South, 1982; Keller et al., 1999). The very dry late spring and summer of 1995 resulted not in post- poned mating, but in a distinctly delayed egg- laying period. This summer drought lasted for six weeks only, and does not seem to have been severe enough to delay egg-laying to the following spring, which is possible in A. lusitanicus (Chevallier, 1971), and has also been observed in A. intermedius by South (1989). But the drought in July 1996 may have caused the gap in egg-laying observed that year. Despite a mild autumn, which is known to accelerate egg-development (Kaiser et al., 1993), the hatching of the juveniles was de- layed compared with 1996, as egg-laying also started later in 1995. Late hatching is defi- nitely disadvantageous for the new genera- tion, as they are not able to start feeding until very late in the year, and thus cannot accu- mulate reserves for the winter. This initial dis- advantage could be overcome by a winter with permanent snow-cover, and thus protec- tion against the low temperatures, followed by a moist and relatively warm spring (as in 1996) with temperatures around optimal lev- els for arionid slugs (Chevallier, 1971; Hom- may, 1994). Such conditions would clearly favour survival and development of young hatchlings and eggs for the next generation. Control Control agents are normally applied when pest slugs are just about to reach maturity and are at a stage when they have the greatest potential for causing damage to crops; not only are they about to reproduce, but it is also known that they require considerably more food during this period of their life cycle (Rollo, 1988). However, in some cases, it may be more appropriate to target the young hatch- lings, as in the case of the parasitic nematode Phasmarhabditis hermaphrodita (Wilson et LIFE CYCLE 8 POPULATION DENSITY OF A. LUSITANICUS 31 al., 1993). This biological control agent has successfully been used against all stages of Deroceras reticulatum, but is known to be in- effective against adults of A. lusitanicus (Glen et al., 1966; Speiser, unpublished); it will, however, attack young hatchlings of this species (Grimm & Hass, in prep.). Thus, the knowledge of the life cycle of A. lusitanicus in central Europe provided by the present study will enable the accurate targeting of particular stages in the life history of A. lusitanicus, and therefore allow a more effective application of control methods in the future. ACKNOWLEDGEMENTS The field work was supported by a research project of the Austrian Federal Ministeries of Science & Transport (GZ 30.505/2-1V/8b/94) and of Agriculture & Forestry (No. L921/94), and publication by a project (GZ 30.686/1- ША/4а/98) of the Austrian Federal Ministry of Science & Transport. | would like to thank Dr. S. E. R. Bailey and Dr. P. B. Mordan for their critical and most helpful reviews of earlier drafts of this manuscript, and Mag. H. Unter- steiner for statistical advice. LITERATURE CITED BARNES, H. F. & J. W. WEIL, 1944, Slugs in gar- dens: their numbers, activities and distribution. Part |. Journal of Animal Ecology, 13: 140-175. BEGON, M., 1979, Investigating animal abundance: capture-recapture for biologists. Arnold, London, England. 97 pp. BEYER, W. N. & D. M. SAARI, 1978, Activity and ecological distribution of the slug Arion subfuscus (Stylommatophora: Arionidae). American Mid- land Naturalist, 100: 359-367. BURENKOV, M. 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SOUTH, A., 1982, Acomparison of the life cycles of Deroceras reticulatum (Muller), and Arion inter- medius (Normand) (Pulmonata: Stylommato- phora) at different temperatures under laboratory conditions. Journal of Molluscan Studies, 48: 233-244. SOUTH, A., 1989, Acomparison of the life cycles of the slugs Deroceras reticulatum (Müller) and Arion intermedius (Normand) on permanent pas- ture. Journal of Molluscan Studies, 55: 9-22. VON PROSCHWITZ, T., 1994, Iberiaskogsnegl — en art pa spredning i Norge. Ettertrykk fra Fauna, 47(3), 195-203. VON PROSCHWITZ, T., 1997, Arion lusitanicus Mabille and A. rufus (L.) in Sweden: A compari- son of occurance, spread and naturalization of two alien slug species. Heldia, 4, (5): 137-138. WEBLEY, D., 1964, Slug activity in relation to weather. Annals of Applied Biology. 53: 407-414. WILSON, M., D. M. GLEN & S. K. GEORGE, 1993, The rhabditid nematode Phasmarhabditis her- maphrodita as a potential biological control agent for slugs. Biocontrol, Science and Technology, 3: 503-511. WOLDA, H. & D. A. KREULEN, 1973, Ecology of some experimental populations of the landsnail Cepaea nemoralis (L.).I. Production and survival of eggs and juveniles. Netherland Journal of Zo- ology, 23: 168-188. YOUNG, A. G., G. R. PORT, A. D. CRAIG, D. A. JAMES & T. GREEN, 1996, The use of refuge traps in assessing risk of slug damage: a com- parison of trap material and bait. In: 1. Е. HENDER- SON, eds. Slug and snail pests in agriculture, BCPC Monograph, 66: 133-140. YOUNG, A. G., G. R. PORT & D. B. GREEN, 1993, Development of a forecast of slug activity: valida- tion of models to predict slug activity from meteo- rological conditions. Crop Protection, 12: 323- 236. Revised ms. accepted 18 July 2000 MALACOLOGIA, 2001, 43(1-2): 33-57 THE UNCERTAIN HISTORY OF LAND SNAILS ON BARBADOS: IMPLICATIONS FOR CONSERVATION Ronald Chase! & David G. Robinson? ABSTRACT There is a growing need to document and conserve molluscan biodiversity, both for scholarly reasons and for public benefit. While the pursuance of these goals necessarily relies on histori- cal records, the accuracy of such records is often taken for granted. We analyzed six previously published lists of land snails on the island of Barbados, and we compared them with results from our own field survey and our study of institutional collections. The current fauna contains six en- demic species. Another endemic, Bulimulus fuscus, is probably extinct, and the status of two ad- ditional endemics, Lucidella barbadensis and Pseudopineria barbadensis, is unknown. Our total of 22 resident species is close to the total number of confirmed species collected by three ear- lier workers dating from 1862, but there are considerable differences in the four lists. When any one list is compared with any one of the others, each contains from one to six species that are absent from the other. Altogether, we confirm 31 species as present on Barbados at some time in the period 1862 to the present, compared with a total of 58 species reported by earlier work- ers. Our analysis allows us to confirm just 23 of the 37 species reported by Brown in 1903, whose list is the basis for a widely consulted conservation reference. By examining institutional collec- tions and tracking down all pertinent literature, we discovered numerous errors of identity, local- ity and taxonomy. Adding to the task facing modern workers is the finding that at least 136 species names have been used to refer to 38 valid taxa, as a result of misidentifications, syn- onyms and genus/species combinations. We conclude that indications of historical trends in snail diversity should be treated with caution until critically evaluated. Key words: Barbados, biodiversity, conservation, land snails, taxonomy. INTRODUCTION The goals of biological conservation now extend to land snails (van Bruggen et al., 1995; Killeen et al., 1998), and studies of the diversity of land snail populations, while for- merly of interest only to biologists, are in- creasingly being used to document habitat degradation (Wareborn, 1992; Gascoigne, 1994; Getz & Uetz, 1994; Graveland et al., 1994). For all these broadening purposes, it is generally recognised that historical records of fauna in a given locality are crucial, yet little consideration has been given to their accu- racy. There exist many records collected not by trained professionals but by amateurs who sometimes lack adequate knowledge or ex- perience. As Willing (1998) has pointed out, we should be grateful to the Conchological Society of Great Britain and Ireland, which has, since its founding in 1876, enforced pro- tocols to ensure authenticity. However, organ- isations such as this do not exist in other lo- calities, and even professional malacologists make unwitting errors. Policymakers as well as scholars often refer to published sum- maries of faunal lists (e.g., Groombridge, 1992) without examining the factual accuracy of the data included therein. Thus, although snails provide good historical records be- cause their shells have been collected for centuries, errors in reporting the data can lead to false interpretations and/or misguided con- servation efforts. There are many sources of error in faunal lists. To begin, surveys of snails in any geo- graphic region must rely on samples collected at selected sites, which necessarily give only estimates of true distributions, and for which the optimal method remains controversial. Identification of species by shell morphology alone is often impossible and, when it is pos- sible, genuine expertise is required. Because species names have changed so often, con- fusion can arise when two collectors or com- mentators apply different names to the same taxon. Voucher specimens are often missing or inaccessible. In addition to these problems, ¿Department of Biology, McGill University, Montréal, Québec H3A 1B1, Canada; ronald.chase @ mcgill.ca Research Associate, Department of Malacology, Academy of Natural Sciences, Philadelphia, Pennsylvania 19103, USA 33 34 CHASE 8 ROBINSON early workers sometimes reported second- hand information of dubious validity. In this paper, we critically examine the his- torical record of land snails on Barbados. Our purpose was not to conduct a definitive sur- vey of the extant fauna, but rather to assess the accuracy and reliability of previous rec- ords. Small islands have traditionally been favoured locations for malacological investi- gations because of their manageable size and geographical isolation (Peake, 1981; Solem, 1984). As a consequence, there is a fairly good record for Barbados, both in the litera- ture and in institutional collections. Our con- cern in this paper is the extent to which errors and ambiguities from the past may cause dif- ficulties for the modern investigator. The historical record of land snails on Bar- bados begins with Griffith Hughes (1750), who included a brief description of three snails in his account of the island’s natural his- tory. Specifically, he mentioned the “land snail . generally of an ash-colour, or black” (possibly Pleurodonte isabella), the “small spiral snails . . . not above half an inch long, very slender, and sharp-pointed” (possibly one of the subulinids), and the “dung-snail . . . very small, and resembles a crusty wart” (possibly Streptaxis glaber). The first mollus- can survey was by Thomas Bland, who lived and collected in Barbados from 1842 to 1849 (Abbott, 1973). Portions of Bland’s material were deposited in ANSP, BMNH and MCZ.? It is unclear from Bland’s publication (Bland, 1862) how many of the specimens he himself collected. He notes that some specimens were collected by Rev. J. Parkinson and Mr. Gill. Other specimens were likely collected by the governor of Barbados, Rawson W. Raw- son, or his agents (records found in the MCZ). Next, in the period 1872-1876, Phillip Car- penter deposited material deposited in the RM. The Carpenter material was collected by Governor Rawson or his agents (letters in the ‘Institutional acronyms: ANSP Academy of Natural Sciences, Philadel- phia, Pennsylvania, USA BMNH Natural History Museum, London, Eng- land, UK FLMNH Florida Museum of Gainesville, Florida, USA FMNH Field Museum of Natural History, Chicago, Illinois, USA MCZ Museum of Comparative Zoology, Harvard Te: Cambridge, Massachusetts, USA RM Redpath Museum, McGill University, Mon- tréal, Québéc, Canada Natural History, McGill University Archives). Kobelt (1880) published a list of species from Barbados based on Bland's list, but with the addition of Hyalina incisa (Pfeiffer). Colonel H. W. Feilden collected shells from Barbados during 1888 and 1889, and some of his material was deposited at ANSP and BMNH. A description was published by Smith & Feilden (1891), based on Feilden’s collection. W. H. Rush was a member of a malacological expedition to a number of Caribbean islands that included a stopover at Bridgetown, Barbados (Rush, 1891). Some of Rush’s material is in the ANSP collection. The Barbados snails were described by L. B. Brown (1903), who de- posited some of his material in the ANSP and the BMNH. Brown also referred to “a type col- lection of nearly all the species enumerated” that had “been placed in the recently-formed Museum attached to the Barbados Natural History Society.” Unfortunately, there is today no trace of such a collection at that institution. The most recent records are by J. B. Hender- son (1919), the malacologist aboard a zoo- logical expedition to the Caribbean mounted by the University of lowa. METHODS Museum Collections Several major institutional collections (iden- tified in footnote 1) were surveyed to locate material deposited by earlier workers. We at- tempted to confirm, or in some cases re-iden- tify, the snails that had been collected from Barbados over the last 150 years. Additional material found in these museum collections, deposited by collectors other than those men- tioned in the Introduction and in Table 1, pro- vided further confirmations for certain taxa. The malacological literature was also sur- veyed, in particular for taxonomic works relat- ing to species represented, or reportedly rep- resented, in Barbados. Field Survey Surveys were conducted by R. Chase in December 1993, December 1994, and March 1996. D. Robinson conducted a survey in Oc- tober 1997. The survey sites are shown in Fig- ure 1, and descriptions are given below. Most sites were examined at least twice, for a total of 2-12 hours per site. All sites were exam- ined at least once during rainfall or immedi- LAND SNAILS OF BARBADOS 35 12 10 O Speightstown O Bridgetown 4 km Elevation >180 m 59°30' W 17,18 [3.153 N 16 e 20,21 11 19 22 14 FIG. 1. Map of Barbados showing locations of sampling sites. ately thereafter. The sites were chosen to pro- vide a fair representation of the island’s geog- raphy and to maximise the total species count. Soil samples where taken at some lo- calities (Stations 1, 2, 7 and 9) where there appeared to be greater potential for small species (loose soil with considerable organic and calcareous particulate matter) and exam- ined under a binocular microscope. Cursory examinations of numerous other sites failed to produce any species additional to those found at the listed sites. The southern portion of the island was surveyed less than other regions because it is heavily populated, relatively dry, and low Iying. It is also widely cultivated with sugar cane, and the cane fields are treated with pesticides. Coordinates were taken using a hand-held GPS (Eagle Explorer), but in in- stances where this was not possible (for ex- ample the presence of the tree canopy block- 36 CHASE 8 ROBINSON ing satellite readings), coordinates were take from topographic maps. Survey Sites de 10. Vie 12: 13. 14. Grounds of Bellairs Research Institute (McGill University), Holetown, Parish of St. James [13711'33"N, 59°38'21°W — alt. 1 m]. Mahogany stand on the grounds of Porter's House, and adjacent wooded gully, Holetown, Parish of St. James [13° 11'44”"N, 59°38'18’W —alt. 2-4 m]. This locality is specifically mentioned by Brown (1903) as one of his collecting sites. Also known as Porter’s Estate or Porter’s Wood. Partially wooded hill northeast of Royal Westmoreland Landscape Garden Cen- tre, Holetown, Parish of St. James [13° 12'01”N, 59°38’04”W — alt. 10 m]. Gully east of Royal Westmoreland Land- scape Garden Centre, Holetown, Parish of St. James [13°12'01”М, 59°38’04”W — alt. 3 m]. Jack-in-the-Box Gully, Parish of St. Thomas. Located approximately 2 km south of Welchman Hall Gully [geodesic coordinates unavailable]. Edge of sugar cane field, adjacent to St. Thomas Church, Parish of St. Thomas [13%11'07”N, 59°36’46”W — alt. 115 m]. Welchman Hall Gully, Parish of St. Thomas [13°11’44”М, 59%34'37"W to 13°11'17”М, 59°34’'34’W —alt. 240-270 m]. Unnamed gully between Welchman Hall Gully and Lion Castle, Parish of St. Thomas [13°11'44’N, 59°34’35’W - alt. 240 т]. Dry coastal forest ~200 m south of Har- rison Point Lighthouse, Parish of St. Lucy [13°18'23’N, 59°38’58”W — alt. 30 m]. Steep rocky slope beneath Harrison Point Lighthouse, Parish of St. Lucy [13° 18’31°N, 59°38’'55°W —alt. 15 т]. East Coast from Bathsheba, Parish of St. Joseph, south to the Congor Rocks, Parish of St. John [geodesic coordinates unavailable]. Friendship, halfway between Crabhill Po- lice Post and Hannays intersection, Parish of St. Lucy [13°18’32”М, 59°37° 45’W —alt. 45 т]. Andromeda Botanic Gardens, Bath- sheba, Parish of St. Joseph [13°12'25’N, 59°31’04”W — alt. 60-90 т]. Ashford plantation, Parish of St. Philip iS: 16. Live 18: 19: 20: 2% 22: 23. 24. [geodesic coordinates unavailable]. Courtesy of Dr. Joan Marsden. 100 m southeast of Mullins Bay, Parish of St. Peter [13°14’00”N, 59°38’26”W — alt. 2 mi. Turner's Hall Woods [Scotland District], Parish of St. Andrew [13°13’23”М, 59°35’ 60”W —alt. 150-180 т]. Although these woods are often referred to as the only virgin stand left on the island (Carrington, 1993), the abundance of introduced snails would seem to indicate that the fauna, at least, is not as “virgin” as others have suggested. Barbados Wildlife Reserve, Parish of St. Andrew [13°16'04”М, 59°35'37’W —alt. 220-240 т]. Grenade Hall Signal Site, Parish of St. Andrew [13%16'05”N, 59°35’33’W — alt. 255]. Roadside, between Malvern House and adjacent sugar cane field, Parish of St. Joseph [13°11’43”М, 59°31'30’W —alt. 275 т]. Woods alongside sugar cane fields of Malvern Plantation, on top of Hackleton's Cliff, Parish of St. Joseph [13°11'58’N, 59°31'21”W — alt. 300 т]. Relatively undisturbed forest, beneath Hackleton’s Cliff, adjacent to Malvern Plantation, Parish of St. Joseph [13°11’ 58’N, 59°31'21”W — alt. 290 m]. This site represents probably the least altered habitat encountered during the study. The absence of any introduced snails, with the occurrence of only three endemic species (Helicina fasciata substriata Gray, Brachypodella costata (Guilding), and Pleurodonte isabella (Férussac)), sug- gests that further exploration here for other endemics would be worthwhile. Grounds of Villa Nova, Parish of St. John [13°10'57’N, 59°31’32”W — alt. 240 т]. Oxnards Crescent, Parish of St. James [geodesic coordinates unavailable]. Courtesy of Dr. Angela Fields. Town of Valley, Parish of St. George [ge- odesic coordinates unavailable]. Cour- tesy of Dr. Angela Fields. RESULTS Survey Results Compared with Previous Records The results of our survey, together with those of earlier workers, are shown in Table 1. A taxonomy of the species is given in the sec- LAND SNAILS OF BARBADOS 37 tion below. For convenience, Carpenter is re- ferred to as an “author” even though he did not publish his findings; his records are dated by the year (1876) in which he last received specimens from Barbados. We found a total of 22 species of land snails. Three of these are new records for Barbados, namely Happiella cf. decolarata, Luntia insignis and Zachrysia provisoria. Hap- piella cf. decolorata and Luntia insignis may be new to the island or they may have been missed by earlier workers because they are uncommon and relatively inconspicuous. Zachrysia provisoria is particularly common in lowland parts of the island, especially along the East Coast where it is the dominant snail species in some areas. It is an introduced species not previously reported as a member of the Barbadian fauna. We failed to find specimens of seven species for which there are either confirmed records for Barbados or unconfirmed records but with a likelihood of occurrence based on distributions on nearby islands (Table 1). Three of the missing species are endemics. Following Breure (1974), it appears that Bu- limulus fuscus is extinct. However, further dili- gent search may yet turn up Lucidella bar- badensis and Pseudopineria barbadensis. Although the 22 species found in our survey corresponds fairly closely to the total numbers of confirmed species found by the most pro- ductive of earlier workers (Bland: 21; Smith & Feilden: 19; Brown: 23), an analysis of the records shows that the composition of the fau- nal lists has undergone constant change. Table 2 shows the number of species reported by any one worker but not by another, which is a measure of faunal instability and/or inac- curate reporting. If the analysis is restricted to the present study plus the three most produc- tive earlier workers (bold font in Table 2), an average of 3.58 species is reported by one worker but not by another (mean of all bolded numbers; range, 1-6). Examination of the row totals reveals no clear historical trend, sug- gesting a constant rate of novel observations. Similarly, there is no clear historical trend in the column totals, suggesting that the authors worked with the same degree of accuracy. In reviewing the results from earlier sur- veys, many errors or ambiguities were dis- covered. Several categories of uncertain records are indicated in Table 1, and a nu- merical summary is given in Table 3. The sources of uncertainty include misidentifica- tions, unverified records, and incomplete naming. Further details are provided in the taxonomic section below. Overall, we are un- able to confirm about one-fifth of all previous records. Taxonomy All species reported from Barbados are in- cluded below, whether from the faunal studies conducted by earlier workers, or from mention of individual species in taxonomies of particu- lar genera or families. Specimens or lots from major institutional collections were examined. The examined specimens comprised voucher materials from earlier workers and any other lots specifically with locality data for Barba- dos. An essential synonymy is provided for each taxon, meaning that the treatment is not exhaustive for species other than the Barba- dos endemics. Species citations are given as used by the original authors, plus any locality data provided. Our intention is to provide suf- ficient detail to convey the intricacy of the his- torical record.” Family HELICINIDAE Helicina fasciata substriata Gray, 1824 Helicina substriata Gray, 1824: 66, pl. 6, fig. 4; Bland, 1862: 351; Gibbons, 1879: 134; Kobelt, 1880: 284; Smith & Feilden, 1891: 256; Brown, 1903: 271 Holotype: not found Helicina Occidentalis Guilding, 1828a: 529 (partim: Barbados only) Helicina conoidea Pfeiffer. Sowerby, 1864: pl. 270 (Helicinidae pl. 5), figs. 168, 169; 1873: pl. 6, fig. 49; Smith & Feilden, 1891: 256, non Pfeiffer, 1854 Helicina convexa Pfeiffer. Rush, 1891: 67, non Pfeiffer, 1849 Helicina fasciata substriata Gray. Wagner, 1911: 334, pl. 67, fig. 6-7; Henderson, 1919: 95-96; Pilsbry, 1930: 229 Distribution: Subspecies (variation ?) H. f. substriata Gray appears to be restricted to Barbados. The typical subspecies, H. f. fasci- ata, has a wider distribution in the Lesser Antilles. Status in Barbados: Locally abundant. Material Collected From: Sites 2, 3, 4, 9, 10, 20, and 21. Additional Material Examined: ANSP 85450 (12 specimens /egit Brown); BMNH 1998103 ?legit = collected by; ex = from the collection of 38 CHASE & ROBINSON TABLE 1. Historical record of species diversity and results of the present survey. Symbols indicate the degree of authen- ticity of the individual records. Only valid species names are listed. In many cases, synonyms were used in the original records, for details of which see the taxonomy section in Results. Records in section e may also appear under the correct name in sections a—d. Column counts for sections a—d include only “X” and “x” symbols. (a (b) Lesser Antillean and/or South American Taxa (8) Cecilioides consobrinus minutissima (Guppy, 1868) Smith & Bland Carpenter Feilden Rush Brown Henderson Present 1862 1876 1891 1891 1903 1919 Study ) Endemic Taxa (9) Brachypodella costata (Guilding, 1828) Bulimulus fuscus Guilding, 1828 Glyphyalinia barbadensis, new name Helicina fasciata substriata Gray, 1824 Lucidella barbadensis (Pfeiffer, 1854) Pleurodonte isabella (Férussac, 1822) Pseudopineria barbadensis Krauss, 1996 Succinea barbadensis Guilding, 1828 Truncatella barbadensis Pfeiffer, 1857 Gastrocopta barbadensis (Pfeiffer, 1853) Happiella cf. decolorata (Drouét, 1859) Luntia insignis Smith, 1898 Megalobulimus oblongus (Muller, 1774) Miradiscops implicans (Guppy, 1868) Orthalicus maracaibensis subpulchella (Pilsbry, 1899) Streptaxis glaber (Pfeiffer, 1849) (c) Pan-Caribbean Taxa (5) Bulimulus guadalupensis (Bruguiere, 1789) Cecilioides aperta (Swainson, 1840) Lacteoluna selenina (Gould, 1848) Lucidella plicatula (Pfeiffer, 1849) Zachrysia provisoria (Pfeiffer, 1858) (d) Pan-Tropical Taxa (8) Allopeas gracile (Hutton, 1834) Allopeas micra (d'Orbigny, 1835) Beckianum beckianum (Pfeiffer, 1846) Bradybaena similaris (Rang, 1831) Gulella bicolor (Hutton, 1834) Leptinaria lamellata (Potiez & Michaud, 1835) Opeas hannense (Rang, 1831) Subulina octona (Bruguiere, 1792) Total number of confirmed taxa: (e) Dubious or Unrecognizable Taxa (12) Bulimulus diaphanus fraterculus (Potiez & Michaud, 1835) Bulimulus tenuissimus (d'Orbigny, 1835) Cecilioides acicula (Müller, 1774) Cerion uva (Linné, 1758) Gastrocopta pellucida (Pfeiffer, 1848) Lacteoluna subaquila (Shuttleworth, 1834) Lacteoluna turbiniformis (Pfeiffer, 1839) Lucidella rugosa (Pfeiffer, 1839) Opeas sp. Polydontes perplexa (Férussac, 1821) Pleurodonte dentiens (Férussac, 1821) Succinea bermudensis Pfeiffer, 1857 Truncatella sp. xx A + +: D xx X XX X X XX x x = хх x KK x x 2? # x XX XXXX XX | O OS x Xx Xx x O os os Vv | MN 2х NI E EC ххх Confirmed taxa (seen): species collected and reported by authors, and confirmed by the existence of corresponding specimens in insti- tutional collections. Confirmed taxa (unseen): species collected and reported by authors; specimens not found in institutional collections but the record is confirmed by ourselves or other workers. Attributed taxa: species reported by authors, but only by attribution to earlier publications. Unconfirmed taxa (likely): species collected and reported by authors; specimens not examined by us; possible occurrence on Barbados based on distribution patterns on nearby islands and on the South American mainland. Unconfirmed taxa (unlikely): species collected and reported by authors, but unlikely to be part of the Barbados fauna based on pub- lished accounts of geographic distribution. Dubious taxa (locality): species reported with correct identification based on our own examination of the same specimens, but with dubi- ous or incorrect locality data. Dubious taxa (identification): species reported with suspected incorrect identification based on our own examination of the same spec- imens. Unrecognizable taxa: taxon name given incompletely by authors. LAND SNAILS OF BARBADOS 39 TABLE 2. Matrix of discrepancies between reports of confirmed species. Numbers represent species reported by author(s) listed at the left but not reported by author(s) listed at the top. Bold is used to highlight studies that reported approximately the same number of total species (range, 19-23). Smith € Bland Carpenter Feilden Bland = 12 4 Carpenter 0 = 0 Smith & Feilden 2 11 = Rush 2 5 1 Brown 3 115 5 Henderson 2 5 0 Present study 5 18 6 COLUMN TOTALS 14 63 16 Present ROW Rush Brown Henderson Study TOTALS 13 1 10 5 45 2 1 2 1 6 10 1 6 3 33 = 0 2 0 10 11 = 10 4 48 4 0 E 0 11 12 4 10 ar 52 52 7 40 13 TABLE 3. Species uncertainty. Numerical summary of the degree of authenticity of faunal studies. The char- acterization of taxa is taken from Table 1, with the same definitions of categories as given there. Confirmed taxa Unconfirmed taxa Dubious taxa Attributed Unrecognizable Seen Notseen Likely Unlikely ID Locality taxa taxa (X) (x) (P) (u) (?) (22) (-) (#) Bland, 1862 11 11 3 Carpenter, 1876 8 Smith & Feilden, 3 16 1 1 3 1891 Rush, 1891 3 7 1 1 Brown, 1903 20 3 2 5 5 Henderson, 1919 12 1 Present study 22 (28 specimens /egit Brown); BMNH living in more humid conditions at higher ele- 1888.8.7.93-105 (13 specimens _ legit vations than the smaller form that predomi- Feilden); FLMNH 185624 (4 miles east of Holetown, St. James —19 specimens); MCZ 90491 (24 specimens legit Rawson ex Bland); MCZ 90492 (27 specimens legit Rawson ex Bland; MCZ 318942 (specimens /egit Rawson ex Binney); MCZ 107933 (Blowers —2 speci- mens legit Kugler); MCZ 83527 (Bridgetown — 1 specimen /egit Garman). Discussion: This taxon belongs to the Heli- cina fasciata complex, the typical subspecies having been described from Puerto Rico and being distributed throughout the Lesser An- tilles, as well as on some western Caribbean islands. Minor differences in island popula- tions have led to a number of names being pro- posed, and only a taxonomic work beyond the scope of this study would determine their va- lidity. Pilsbry (1930) considered that Barbados specimens match the description and figure of Helicina substriata Gray, and on that basis he designated Barbados as the type locality for this taxon without designating a lectotype. The specimens of this species on Barbados range considerably in size, larger specimens nates in drier areas, especially along the northwestern coast. This arboreal species was collected at many of the localities stud- ied, and is particularly abundant where there is less human disturbance, crawling on leaf surfaces, tree trunks and aerial tree roots. Lucidella (Poeniella) barbadensis (Pfeiffer, 1854) Helicina Barbadensis Pfeiffer, 1854: 60 (“Bar- badoes”); Bland, 1862: 351; Kobelt, 1880: 284; Smith & Feilden, 1891: 256; Brown, 1903: 266, 271 Type material: BMNH 1998024 (Cuming col- lection) Helicina conoidea Pfeiffer, 1854: 53 (“Barba- does, West Indies”) Possible type — BMNH 1998025 (Cuming col- lection) Not Helicina conoidea Pfeiffer. Sowerby, 1864: pl. 270 (Helicinidae pl. 5), figs. 168, 169; 1873: pl. 6, fig. 49 (=Helicina fasci- ata var. substriata Gray, 1824) 40 CHASE 8 ROBINSON Lucidella holoserica Wagner, 1910: 350-351, pl. 69, fig. 16-19 (“die Insel Barbados”) Lucidella (Poeniella) barbadensis (Pfeiffer). Baker, 1962: 18) Distribution: Endemic to Barbados. Status: Extinct? Material Examined: ANSP 14916 (4 speci- mens /egit Bland); ANSP 14926 (14 speci- mens [labelled as “Helicina conoidea Pfr.?”] legit Shuttleworth); ANSP 85468 (12 speci- mens /egit Brown); BMNH (5 specimens legit Brown); BMNH 97.10.12.3-7 (Hackleton's Cliff-5 specimens legit Pavey); MCZ 90489 (2 specimens ex Bland); MCZ 107932 (Blowers — 1 specimen legit Kugler). Discussion: The identity of Helicina conoidea has been problematical since its first descrip- tion. Pfeiffer's taxon is a synonym of Lucidella barbadensis, described by him in the same year in the same paper. An examination of the material in the BMNH used by Pfeiffer to de- scribe the two taxa shows them to be within the range of variation of a single species. Sowerby (1873) in his Helicina monograph il- lustrated a totally different species, a fact that was somewhat acidly pointed out by Bland (1857: 247), although Bland himself seemed confused as to the identity of the taxon (see under Lucidella plicatula below). Smith & Feilden (1891) merely referred to the speci- mens in the Cuming collection. Brown even expressed doubt as to the occurrence of He- licina conoidea Pfeiffer on Barbados; he may indeed have been considering H. conoidea of Bland, as Brown did collect specimens of L. barbadensis from Porter’s Wood. This species was reported as uncommon in the past, and no specimens were encoun- tered during our survey. Lucidella (Poeniella) plicatula (Pfeiffer, 1849) Helicina plicatula Pfeiffer, 1849: 123 (“Mar- tinique”) Helicina conoidea Pfeiffer. Bland, 1862: 351; Kobelt, 1880: 284, non Pfeiffer, 1854 Helicina rugosa (Pfeiffer). Brown, 1903: 266, 271, non Pfeiffer, 1839 Distribution: Hispaniola; Puerto Rico and the Lesser Antilles. Status in Barbados: Extirpated? Material Examined: ANSP 14783 (3 speci- mens /egit Bland); MCZ 318945 (10 speci- mens ex Bland); MCZ 107931 (Blowers — 1 specimen /egit Kugler). Discussion: The only known Barbados spec- imens of this widely distributed West Indian species available for inspection were col- lected by Bland, but misidentified as Helicina conoidea Pfeiffer. Brown’s (1903) reference to the Cuban taxon Helicina rugosa Pfeiffer, de- scribing it as a “smaller shell” and “identified by the very strong diagonal striation,” clearly is a reference to this species, and establishes its occurrence in Porter’s Wood (St. James) at the time. Family TRUNCATELLIDAE Truncatella barbadensis Pfeiffer, 1857 Truncatella Barbadensis Pfeiffer, 1857: 337 (‘Island of Barbadoes”); Bland, 1862: 351; Kobelt, 1880: 284; Smith & Feilden, 1891: 255-256; Brown, 1903: 271; Torre, 1960: 87 Types: BMNH (Cuming Collection)— lost. ? Truncatella sp. Henderson, 1919: 96 Truncatella (Truncatella) bilabiata Pfeiffer. Clench & Turner, 1948: 153-154 (partim) Truncatella sp. nov. Rosenberg, 1996: 682- 693 Distribution: Endemic to Barbados. Status: Locally abundant. Material Collected From: Sites 9 and 10. Additional Material Examined: ANSP 85463 (8 specimens legit Brown); ANSP 397286 (Harrison Point Lighthouse — 100+ specimens legit Rosenberg); BMNH 88.8.7.106-17 (12 specimens legit Feilden); BMNH 88.8.7.84- 92 (9 specimens legit Feilden); BMNH 1998105 (8 specimens legit Brown); FLMNH 271905 (Harrison’s Cave —6 specimens legit F. G. Thompson); Discussion: Although Pfeiffer described Trun- catella barbadensis as a distinct species from Barbados, Clench & Turner (1948) syn- onymized it with the marine littoral and more widespread T. bilabiata (Pfeiffer), and despite Torre's (1960) arguments that the Barbadian species should be considered distinct, this has generally been followed by workers ever since. Rosenberg (1996) recognized that the form he found on Barbados was quite differ- ent from other known Truncatella species from elsewhere in the Caribbean, suggested LAND SNAILS OF BARBADOS 41 that it was a new species, but he did not de- scribe it. Feilden's material in the British Museum and Brown's material at both the British Museum and the Academy of Natural Sciences clearly are this species. Unfor- tunately, the matter cannot be laid to rest con- clusively, because Pfeiffer’s type material is lost, and a neotype will have to be designated in order to define the taxon. Henderson’s (1919) reference to a Truncatella species could be this taxon, or it could be one of the marine littoral Truncatella species that are also part of the Barbados fauna. This species lives primarily in the leaf litter of coastal forest under completely terrestrial conditions at over 30 m altitude, at the top of the coastal cliffs. However, it can tolerate habitats influenced by the marine environ- ment, some individuals living closer to the ocean under stones and boulders in areas somewhat affected by distant sea spray. It does not appear to live in the supralittoral zone environment, with which other species of this genus are normally associated. Family SUCCINEIDAE Succinea barbadensis Guilding, 1828 Succinea Barbadensis Guilding, 1828a: 532; Bland, 1862: 351, table 2; Gibbons, 1879: 132; Kobelt, 1880: 284; Smith € Feilden, 1891: 255; Rush, 1891: 69; Brown, 1903: 270 Succinea Bermudensis Pfeiffer. Bland, 1862: 351, tabl. 2; Kobelt, 1880: 284, non Pfeif- fer, 1857 Succinea sp. Henderson, 1919: 96 Distribution: Barbados; throughout the West Indies? Status in Barbados: Locally abundant. Material Collected From: Sites 3, 4, 7, 9, 13, 16, 18, and 20. Additional material examined: ANSP 10249 (1 specimen /egit Rush); ANSP 85459 (6 specimens /egit Brown); ANSP 332700 (Porter's Estate —1 specimen /egit Hussey); RM 12075 (5 specimens legit Rawson ex Car- penter); BMNH 88.8.7.118-26 (9 specimens legit Feilden); BMNH 1998104 (11 specimens legit Brown); MCZ 90372 (11 specimens legit Rawson ex Bland); MCZ 90373 (5 specimens legit Rawson ex Bland); MCZ 74182 (9 spec- imens legit Rawson); MCZ 318943 (near Gray's Cove, St. Lucy-St. Peter—14 speci- mens /egit Clench). Discussion: A widespread and variable species in Barbados. Despite the variability in shell shape, all the specimens dissected dur- ing this study show an absence of anatomical variation and are considered to represent a single species. Unfortunately, the taxonomy of Caribbean Succinea species is in a state of disarray, little anatomical work having been done to properly define the numerous named taxa. It is therefore impossible to determine the true geographical extent of this species. The name S. barbadensis has been used in the literature for shells collected throughout the West Indies, but those records are here excluded from the synonymy. Family VERTIGINIDAE Gastrocopta barbadensis (Pfeiffer, 1853) Pupa Barbadensis Pfeiffer, 1853: 554 (“in in- sula Barbadoes”); Bland, 1862: 351; Ko- belt, 1880: 284 Pupa Jamaicensis Adams. Bland, 1862: 351; Kobelt, 1880: 284 Pupa pellucida Pfeiffer. Bland, 1862: 351; Gibbons, 1879: 132; Kobelt, 1880: 284; Smith & Feilden, 1891: 254-255; Brown, 1903: 270, non Pfeiffer, 1848 Gastrocopta barbadensis (Pfeiffer). Pilsbry, 1916: 83-85, pl. 18, figs. 1-5 Distribution: Lesser Antilles, Venezuelan off- shore islands and coastal Venezuela. Status in Barbados: Uncommon. Material Collected From: Site 9. Additional Material Examined: ANSP 85464 (11 specimens /egit Brown); ANSP 332704 (Porter’s Estate —2 specimens /egit Hussey); ANSP 328617 (Blowers, St. James —1 speci- men /egit Kugler); MCZ 90573 (Blowers — 1 specimen /egit Kugler). Discussion: Various workers have used three different names for this species, Pupa pellu- cida Pfeiffer, 1848, originally described from Cuba, its junior synonym P. jamaicensis Adams, 1849, from Jamaica, and P. bar- badensis Pfeiffer, 1853, described from Barbados. Smith & Feilden (1891) suggested all were the same species. There is no evi- 42 CHASE 8 ROBINSON dence for the occurrence of Gastrocopta pel- lucida (Pfeiffer) on Barbados; its known distri- bution is from eastern Mexico, the Bahamas Islands and the Greater Antilles, and Ber- muda (Pilsbry, 1916). Feilden collected Pupa barbadensis at Maxwell Hall (Parish of Christchurch), Hussey at Porter's Estate (St. James), and specimens were collected during this study from eastern Parish of St. Lucy. One of the smallest species of the Barbados molluscan fauna, it is easily missed, and may have a wider distribu- tion on the island. Pupisoma (Ptychopatula) dioscoricola (Adams, 1845) Helix dioscoricola Adams, 1845: 16 Distribution: Ubiquitous Americas. Status in Barbados: Rare? Material Examined: FLMNH 119895 (Harri- son’s Cave, St. Thomas 1 specimen /legit Thompson); throughout the Discussion: Although a single specimen of this species has been collected in Barbados in recent years (none was collected during our field study), it is likely to be established in Bar- bados despite the lack of records, in view of its ubiquitous nature in anthropochorous envi- ronments throughout the Americas. It is minute and easily missed. Family CERIONIDAE Cerion uva (Linné, 1758) Turbo uva Linné, 1758: 765 Cerion uva (“Lamarck”). Brown, 1903: 270 Distribution: Endemic to Aruba, Curacao and Bonaire. Status in Barbados: Dubious record. Discussion: Although Brown mentioned the collection of two specimens of this species by a Rev. N. B. Watson, one from St. Peter’s and one from St. Philip’s Parishes, there is no physical evidence that this species, or indeed any species of Cerion has ever been collected from Barbados. No specimens of this species collected from anywhere other than from the Dutch islands off the northern Venezuelan coast are in the ANSP, FLMNH or BMNH col- lections. Family BULIMULIDAE Bulimulus guadalupensis (Bruguiére, 1789) Bulimus Guadalupensis Bruguiére, 1789: 313 Bulimus exilis (Gmelin). Bland, 1862: 351, non Helix exilis Muller, 1774 Bulimulus exilis (Gmelin). Gibbons, 1879: 130; Kobelt, 1880: 284; Smith & Feilden, 1891: 252; Brown, 1903: 267, 269; Hen- derson, 1919: 202 Bulimulus exiles [sic] (Gmelin). Rush, 1891: 69 Bulimulus (Thaumastus) Smith, 1895: 302, 305 exilis (Gmelin). Distribution: West Indies, and northern South America. Status in Barbados: Locally abundant. Material Collected From: Sites 2, 3, 4, 5, 7, 124193 and 22; Additional Material Examined: ANSP 85439 (6 specimens legit Brown); ANSP 85440 (6 specimens /egit Brown); ANSP 85441 (6 specimens legit Brown); ANSP 85442 (8 spe- cimens legit Brown); ANSP 62059 (6 speci- mens /egit Rush); RM 13620 (8 specimens [labelled as Bulimus exilis] legit Rawson ex Carpenter); RM 13623 (St. Thomas Parish —3 specimens [labeled as Bulimus guadalupen- sis] legit Rawson ex Carpenter); FLMNH 113848 (Harrison's Cave, St. Thomas —4 specimens /egit Thompson); MCZ 90167 (100+ specimens —/egit Rawson ex Bland); MCZ 83525 (Bridgetown —2 specimens /egit Garman); МСУ 108728 (St. Joseph’s —4 specimens /egit Cockerell); MCZ 251316 (St. Ann’s Fort, St. Michael /egit Ray & Allen). Discussion: This West Indian species is found throughout Barbados whereever there has been any kind of human activity. Breure (1974) considers this species native to the Windward Group of the Lesser Antilles, so it was probably introduced to Barbados by human commerce. Bulimulus fuscus Guilding, 1828 Bulimulus fuscus Guilding, 1828b: 170 (Bar- bados) Bulimus Barbadensis Pfeiffer, 1854: 61 (“in in- sula Barbados”); Bland, 1862: 351; Pils- bry, 1897: 48-49, pl. 12, fig. 61 Bulimus fuscus (Guilding). Bland, 1862: 351 LAND SNAILS OF BARBADOS 43 Bulimulus fuscus (Guilding). Kobelt, 1880: 284 Bulimulus tenuissimus (Férussac). Smith 4 Feilden, 1891: 252; Brown, 1903: 268 (doubtful occurrence; attributed to earlier workers), non Bulimus tenuissimus d'Or- bigny, 1835 Bulimulus (Bulimulus) fuscus Guilding. Breure, 1974: 38-39, pl. V, figs. 1-4; pl. Vil 89:5: Distribution: Endemic to Barbados. Status: Extinct? Material Examined: BMNH 197454 (lecto- type designated by Breure, 1974); BMNH 197455 (2 paralectotypes); ANSP 3506 (3 specimens legit Bland); ANSP 3507 (1 speci- men /egit Bland); ANSP 3512 (3 specimens legit Bland); ANSP 25612 (8 specimens /egit Bland); RM 13650 (4 specimens legit Rawson ex Carpenter); MCZ 90415 (6 specimens /egit Rawson ex Bland). Discussion: Smith & Feilden (1891) syn- onymized Bulimus barbadensis Pfeiffer [= Bu- limulus fuscus Guilding], which Feilden had collected in Barbados, with Bulimulus tenuis- simus, a superficially similar Brazilian species. Pilsbry (1897: 49) also noted the similarity be- tween the two taxa, but confirmed that the lat- ter was a distinct Brazilian species. Breure (1974) in his treatment of Caribbean Bulimulus suggested that this species may be extinct, as all records are from the nineteenth century. No specimens were collected during this study. Bulimulus diaphanus fraterculus (Potiez & Michaud, 1835) Helix (Cochlogena) fraterculus Férussac, 1821: 54 (“La Guadeloupe”) (nomen nudum) Bulimus fraterculus “Férussac.” Potiez & Michaud, 1835: pl. 13, figs. 7-8; 1838: 141 (“La Guadeloupe”) Bulimus fraterculus “Férussac.” Bland, 1862: 351 Bulimulus fraterculus “Ferussac.” Kobelt, 1880: 284; Smith & Feilden, 1891: 252 Bulimulus diaphanus (Pfeiffer). Pilsbry, 1897: 47 (Barbados record), non Bulimus di- aphanus Pfeiffer, 1854 Bulimulus fraterculus “Ferussac.” Brown, 1903: 269 (doubtful occurrence; attrib- uted to Bland) Bulimulus (Bulimulus) diaphanus fraterculus (Potiez & Michaud). Breure, 1974: 32-34, pl. 3, figs. 6-10; pl. 7, fig. 1 Distribution: St. Martin; Saba; St. Eustatius; Barbuda; Guadeloupe; Barbados? Status: Unknown. Dubious record. Material Examined: ANSP 25609 (2 speci- mens /egit Foderougher). Discussion: Brown (1903) stated that he was unable to confirm the occurrence of this species in Barbados. Breure (1974) in his tax- onomic review of the genus Bulimulus in the Caribbean indicated that the Barbados records of this taxon are “very doubtful,” con- sidering that the records by Bland (1862), and Smith & Feilden (1891) are probably mis- identifications of the polymorphic Bulimulus guadalupensis. However, two specimens in the ANSP collection, reportedly from Barba- dos, are clearly B. d. fraterculus. Whether this indicates it was established in Barbados at one time, or represents an error in locality re- mains uncertain. Plekocheilus aurissileni (Born, 1780) Voluta auris Sileni Born, 1780: 212, pl. 9, figs. 3,4 Plekocheilus (Plekocheilus) aurissileni (Born). Breure, 1975: 73-76, pl. VI, figs. 5-10 Distribution: St. Vincent. Status in Barbados: Dubious record. Discussion: Breure (1975) reported this large and distinctive species being collected from Porter's Wood (north of Holetown, Parish of St. James) in Barbados. This was based on a specimen that was in Alan Solem's private collection but was not collected by him per- sonally (J. Siapcinsky, personal communica- tion), and was deposited in the Field Museum of Natural History, Chicago (FMNH 146430). There is no record of Solem ever having been to Barbados to collect, and presumably he ob- tained the specimen from an unknown collec- tor, together with erroneous locality data. Orthalicus maracaibensis subpulchella (Pilsbry, 1889) Orthalicus zebra (Múller). Smith 8 Feilden, 1891: 253; Smith, 1895: 302, 306-307; Rush, 1891: 69 (Bridgetown); Brown, 44 CHASE 8 ROBINSON 1903: 269, non Buccinum zebra Muller, 1774 Oxystyla maracaibensis var. subpulchella Pilsbry, 1889: 141-142, pl. 28, fig. 38, 39 (Union Island, Grenadines) Oxystyla sp. Henderson, 1919: 95 Oxystyla pulchella (Spix). McGinty, 1939: 7, pl. 2, fig. 9, non Achatina pulchella Spix, 1827 Distribution: Barbados and the Grenadines. Status in Barbados: Common. Material Collected From: Sites 1, 3, 4, 5, 7, Abi and 22: Additional Material Examined: ANSP 89699 (nr. Bridgetown—1 specimen /egit Clapp); ANSP 227834 (Bridgetown—2 specimens legit Bales); ANSP 303292 (St. John’s Wood — 18 specimens legit Jackson); FLMNH 113849 (Harrison’s Cave, St. Thomas—1 specimen legit F. G. Thompson); FLMNH 113851 (Welchman Hall Gully—4 specimens); FLMNH 109541 (near Bank Hall —5 speci- mens; FLMNH 177917 —St. Peter mangrove plantation); MCZ 251304 (Barbados Museum, St. Michael legit Ray 8 Allen); MCZ 21085 (near Bridgetown —3 specimens /egit Kugler); MCZ 142231 (St. John’s Church, St. David — 49 specimens /egit Howland); MCZ 108953 (Hastins —36 specimens /egit Howland). Discussion: The taxonomic status of this common Barbados tree snail is uncertain, al- though it most closely resembles Orthalicus maracaibensis subpulchella (Pilsbry, 1889), from the Grenadines (typical maracaibensis is from the mainland of Venezuela). Henderson (1919) also recognized the Barbadian form to be “more nearly related to a South American group” as opposed to O. undatus jamaicensis Pilsbry, 1889. Pilsbry (1889) had placed refer- ences to Orthalicus zebra (Muller) (including that of Smith & Feilden, 1891) in synonymy with Oxystyla undatus var. jamaicensis Pils- bry, 1889, presumably without seeing speci- mens from Barbados (the specimens in the ANSP collection were received after Pilsbry had written the relevant text in the Manual of Conchology). Brown (1903) also discussed the taxonomic uncertainty of this species. The taxonomy of the various Caribbean and main- land species and named forms of Orthalicus is in need of revision. Smith & Feilden (1891) reported that Feilden had “brought a small basket full of [Orthalicus undatus jamaicensis] from Ja- maica to Barbados,” that was then released on Pelican Island, a quarantine station just off Bridgetown. Feilden is quoted as later discov- ering that he “found them in limited numbers already introduced to gardens in the suburbs of Bridgetown.” Doubtless he, too, was misled by the similarity of the two forms. Whether or not the Jamaican taxon has survived some- where in the Bridgetown area is unknown. Smith & Feilden’s, and Rush's records (both 1891) are the earliest records of a species of Orthalicus in Barbados. It would seem that the species had recently been in- troduced from elsewhere, presumably the nearby Grenadines, as the species is now common in trees in inhabited areas and so visible as not to be easily missed. Family UROCOPTIDAE Brachypodella costata (Guilding, 1828) Brachypus costatus Guilding, 1828b: 167 (“in arboribus Barbadensibus”) Siphonostoma costata (Guilding). Swainson, 1840: 168, fig. 22 (non p. 333, fig. 97c, d) Cylindrella costata (Brachypus) (Guilding). Pfeiffer, 1844: 183, pl. 1, fig. 16 only; Bland, 1862: 351; Kobelt, 1880: 284 Cylindrella (Gongylostoma) costata Guilding. Smith 8 Feilden, 1891: 255; Brown, 1903: 270 Urocoptis sp. Henderson, 1919: 96 Brachypodella costata (Guilding). 1904: 78-79. Brachypodella costata form albida Pilsbry, 1904: 79. Pilsbry, Distribution: Endemic to Barbados. Errone- ous records from St. Lucia. Status: Locally abundant. Material Collected From: 2, 7, 8, 9, 13, 19, and 21. Additional Material Examined: ANSP 73226 (10 specimens /egit Bland); ANSP 73228 (8 specimens /egit Bland—types of Brachy- podella costata form albida Pilsbry, 1904); ANSP 73229 (13 specimens legit Bland); ANSP 85458 (13 specimens legit Brown); RM 14180 (7 specimens /egit Rawson ex Carpen- ter); FLMNH 119898 (Harrison's Cave, St. Thomas —12 specimens /egit Thompson); MCZ 171018 (13 specimens legit Rawson ex Bland); MCZ 26922 (4 specimens ex Bland); LAND SNAILS OF BARBADOS 45 MCZ 75023 (Blowers — 100+ specimens legit Kugler). Discussion: This species is common throughout the island, in some areas being abundant, found crawling on tree trunks and other vertical surfaces, as well as on rocks and boulders. Brown (1903) listed this species also being from St. Lucia, but this record refers to the similar Brachypodella tatei (Crosse). Pseudopineria barbadensis Kraus, 1996 Bulimus Viequensis Pfeiffer. Bland, 1862: ЗЭ ре. Pilsbry, 1904: 111=112 pl: 1, fig. 12, only, non Pfeiffer, 1856 Pineria Viequensis var. minor Pfeiffer, 1868: 343 (Barbados) Stenogyra Viequensis (Pfeiffer). Kobelt, 1880: 284, non Pfeiffer, 1856 Pineria viequensis (Pfeiffer). Smith & Feilden, 1891: 253; Brown, 1903: 269, non Pfeif- fer, 1856 Pseudopineria barbadensis Kraus, 109-113, figs. 8, 10 1996: Distribution: Endemic to Barbados. Status: Unknown. Material Examined: FLMNH 50324 (holo- type); FLMNH 180526 (5 paratypes legit J. J. Brown); ANSP 85454 (12 paratypes legit Brown); MCZ 23720 (17 paratypes legit Guppy). Discussion: Brown (1903) reported this species as Pineria viequensis occurring along the coast of the Parish of St. Philip, and Feilden collected it from the Parishes of Christchurch and St. Philip. Kraus (1996) rec- ognized the Barbados records as a distinct species based on museum specimens. No specimens were collected during this study, so its status is unknown. Family SUBULINIDAE Allopeas gracile (Hutton, 1834) Bulimus gracilis Hutton, 1834: 84 (Mirzapoor . . . Futtehpoor Sikra. . . between Agra and Neemuch [India]) Bulimus subula (Pfeiffer). Bland, 1862: 351 Stenogyra subula (Pfeiffer). Kobelt, 1880: 284; Smith & Feilden, 1891: 253-254 Opeas subula (Pfeiffer). Brown, 1903: 270 ? Opeas sp. Henderson, 1919: 96 Distribution: Tropics and subtropics world- wide; some Pacific island groups. Status in Barbados: Uncommon. Material Collected From: Sites 1, 3, and 13. Additional Material Examined: ANSP 3101 (4 specimens /egit Bland); ANSP 85451 (4 spec- imens legit Brown); MCZ 318940 (Blowers — 1 specimen /egit Kugler). Discussion: A worldwide species introduced by humans. Rush’s (1891) reference to a species of Opeas could be either one of the Allopeas taxa listed here or Opeas hannense. It is included here as Allopeas gracile, be- cause it is the largest of the three and the one most likely encountered by Rush. Allopeas micra (d’Orbigny, 1835) Helix (Achatina) micra d’Orbigny, 1835: 9 (Central America) Stenogyra micra (d’Orbigny). Gibbons, 1879: 13 Bulimus octonoides Adams. Bland, 1862: 351 Stenogyra octonoides Adams. Kobelt, 1880: 284; Smith & Feilden, 1891: 254; Rush, 1891: 69 Opeas octonoides Adams. Brown, 1903: 270 Distribution: Mexico to Bolivia, and the West Indies; some Pacific island groups. Status in Barbados: Uncommon. Material Collected From: Sites 3, 7, and 9. Additional Material Examined: ANSP 3107 (21 specimens /egit Bland); ANSP 85460 (4 specimens /egit Brown). Discussion: A synanthropic species spread throughout the Americas and elsewhere. Beckianum beckianum (Pfeiffer, 1846) Bulimus beckianus Pfeiffer, 1846: 82 (Opara) Bulimus Caraccasensis Reeve. Bland, 1862: 351 Stenogyra caracasensis (sic) Reeve. Kobelt, 1880: 284 Stenogyra Beckiana Pfeiffer. Smith & Feilden, 1891: 253; Rush, 1891: 69 46 CHASE & ROBINSON Opeas Beckiana Pfeiffer. Smith, 1895: 302, 309; Brown, 1903: 270 Distribution: South and Central America, and the West Indies; some Pacific island groups. Status in Barbados: Locally abundant. Material Collected From: Sites 1, 2, 3, 7, 9, 19, and 22. Additional Material Examined: ANSP 337234 (1 specimen /egit Bland); ANSP 85461 (12 specimens legit Brown); ANSP 332714 (Porter's Estate —2 specimens /egit Hussey); RM 15346 (8 specimens legit Rawson ex Car- penter); MCZ 27231 (12 specimens legit Rawson); MCZ 136007 (9 specimens ex Bland); MCZ 318944 (near Fresh Water Bay legit Balch); MCZ 90579 (Blowers —3 speci- mens ex Kugler). Discussion: Smith & Feilden (1891) reported this species as being “not very common.” Today the species is particularly widespread, often abundant, especially in disturbed habi- tats. Leptinaria lamellata (Potiez & Michaud, 1835) Achatina lamellata Potiez & Michaud, 1835: pl. 11, figs. 7, 8; 1838: 128 (Hab. ?) Tornatellina Antillarum Shuttleworth. Bland, 1862: 351 Leptinaria antillarum (Shuttleworth). Kobelt, 1880: 284 Leptinaria lamellata (Potiez & Michaud). Smith & Feilden, 1891: 255; Smith, 1895: 302, 309; Brown, 1903: 270 Leptinaria sp. Henderson, 1919: 96 Distribution: Tropics and subtropics world- wide. Status in Barbados: Locally common. Material Collected From: Sites 2, 7, 13, and 19. Additional Material Examined: ANSP 24089 (2 specimens legit Bland); ANSP 85456 (4 specimens /egit Brown); MCZ 90578 (Blow- ers —3 specimens legit Kugler). Discussion: This species is widely distributed on Barbados, although never in great num- bers. It is associated with areas of human dis- turbance. Luntia insignis Smith, 1898 Luntia insignis Smith, (Trinidad) Distribution: Trinidad; Aruba; Saba; Barba- dos. 1898: 28, figs 8 Status т Barbados: Uncommon. Material Collected From: Sites 2, 3, 9, and 13. Discussion: This small subulinid is here re- ported from Barbados for the first time. It was originally known only from Trinidad, until it was reported on Aruba by Wagenaar Hum- melink (1940), and then on Saba by Haas (1962). It is probably distributed throughout the Lesser Antilles. Opeas hannense (Rang, 1831) Hélice (Cochlicelle) hannensis Rang, 1831: 41-42, pl. 3, fig. 8 (Village de Hann sur la presque’ile du Cap-Verd” [West Africa]) Bulimus Goodalli (Miller). Bland, 1862: 351 Stenogyra Goodalli (Miller). Kobelt, 1880: 284; Smith & Feilden, 1891: 254 Opeas goodalli (Miller). Brown, 1903: 270 Opeas ascendens Poey. Brown, 1903: 270 Distribution: Tropical Central America; intro- duced worldwide. Status in Barbados: Rare. Material Collected From: Site 9. Additional Material Examined: ANSP 85462 (4 specimens legit Brown). Discussion: We follow Proschwitz (1994) and Cowie (1997) in using Helix hannensis Rang as senior synonym of Helix goodalli Miller, 1822, and Bulimus pumilus Pfeiffer, 1840. Although Brown (1903) listed both O. goodalli and O. ascendens, a note in the text by E. A. Smith (in Brown, 1903: 270) indicates that they are the same species. Although reported by various other workers on the Barbadian fauna, we found only two specimens of this species at a single locality. Subulina octona (Bruguiere, 1792) Bulimus octonus “Chemnitz” Bruguiere, 1792: 325 (“Les lles Antilles”) Achatina octona (Bruguiere). Bland, 1862: 351 LAND SNAILS OF BARBADOS 47 Stenogyra octona (Bruguiere). Gibbons, 1879: 131; Kobelt, 1880: 284; Smith 8 Feilden, 1891: 253 Subulina sp. Rush, 1891: 69 Subulina octona (Bruguiere). Smith, 1895: 302, 309; Brown, 1903: 270 Distribution: Tropics and subtropics world- wide, as well as in greenhouses in the tem- perate zones of Europe and North America. Status in Barbados: Common, locally abun- dant. Material Collected From: Sites 1, 2, 3, 5, 6, 7, 9.12.1315. 16, 19. 20. and 22: Additional Material Examined: ANSP 85453 (8 specimens /egit Brown); RM 12742 (8 specimens /egit Rawson ex Carpenter); MCZ 27131 (7 specimens —/egit Rawson); MCZ 136006 (9 specimens ex Bland); MCZ 107934 (Blowers —31 specimens ex Kugler). Discussion: One of the commonest and most widespread species on Barbados. Smith & Feilden (1891) reported that “it is very abun- dant throughout the island, and is met with in colonies under stones and rocks.” Specimens are often observed containing a number of white eggs in the last two or three whorls of the shell, even when the shell is only half the length of the largest specimens encountered. It would appear that this snail, as other subu- linids, is capable of reproduction well before its maximum size is reached. Family FERUSSACIIDAE Cecilioides acicula (Müller, 1774) Buccinum acicula Muller, 1774: 150-151 Caecilianella acicula (Miller). Brown, 1903: 266, 270 Distribution: Europe; isolated records of in- troductions to other continents and Pacific is- lands. Status in Barbados: Dubious record. Discussion: Brown (1903) reported collecting a single specimen at Porter's Wood (St. James), together with specimens of C. conso- brinus minutissima (Guppy) (see below). However, he also misidentified specimens of C. aperta, which upon closer examination are Clearly individual variations of C. consobrinus minutissima. It is likely that his determination of this European species was also in error. Cecilioides (Geostilbia) aperta (Swainson, 1840) Macrospira aperta “Guilding” Swainson, 1840: 335, fig. 97e, f Achatina Gundlachi Pfeiffer. Bland, 1862: 351 Stenogyra Gundlachi (Pfeiffer). Kobelt, 1880: 284 Stenogyra Gundlachi “Arango” Smith 4 Feilden, 1891: 254 (attributed to Bland, 1862) Caecilianella aperta “Guilding” Brown, 1903: 269-270 Opeas gundlachi“Arango.” Brown, 1903: 266, 270 (doubtful occurrence; attributed to Bland) Caecilianella gundlachi Pfeiffer. Brown, 1903: 270 Distribution: West Indies. Status in Barbados: Unknown. Material Examined: MCZ 90580 (Blowers — 10 specimens ex Kugler). Discussion: Although Bland (1862) and Brown (1903, in Porter’s Wood listed as C. gundlachi) reported this species from Barba- dos (Smith & Feilden, 1891, merely quoted Bland’s record), we have been unable to find specimens from the island in any of the insti- tutional collections surveyed, although there is no reason to suppose this widely distributed Caribbean species has never been present on the island. Smith (1895) reported it from the neighbouring island of St. Vincent. Brown attributed Opeas gundlachi Arango to Bland. However, it was Smith & Feilden who used this name combination, although they were referring to Bulimus gundlachi Pfeiffer [= Ce- cilioides aperta (Swainson)], a species Brown believed he had collected (see next species). No specimen attributable to this taxon was collected during our survey. Cecilioides (Karolus) consobrinus minutissima (Guppy, 1868) Cæcilioides minutissima Guppy, 1868a: 239 (Trinidad) Ceecilianella minutissima (Guppy). Brown, 1903: 269 Ceecilianella aperta Guilding. Brown, 1903: 48 CHASE & ROBINSON 270, non Macrospira aperta Swainson, 1840 Ceecilianella (Cæcilioides) consobrinus var. minutissima (Guppy). Pilsbry, 1909: 41-42, pl. 5, figs. 83, 85 Distribution: Trinidad; St. Vincent; Barbados. Status in Barbados: Unknown. Material Examined: ANSP 85457 (5 speci- mens [labelled as Ceecilianella minutissima] legit Brown); ANSP 85466 (3 specimens [la- belled as Ceecilianella aperta] legit Brown). Discussion: Pilsbry (1909) recognized the race/variety minutissima from the southern Lesser Antilles as distinct from the typical form. Brown collected two confirmed lots of this species from Barbados, although it was not found by any subsequent workers, includ- ing ourselves. lt is a minute species living cryptically, and as such, it would be premature to conclude that it no longer lives on the is- land. Family STREPTAXIDAE Streptaxis (Streptartemon) glaber (Pfeiffer, 1849) Streptaxis glabra Pfeiffer, 1849: 126 (Demer- ara) Streptaxis deformis (Férussac). Smith & Feilden, 1891: 251; Rush, 1891: 68; Brown, 1903: 268; Henderson, 1919: 95; Pilsbry, 1908: x, pl. 52, fig. 5, non Helix deformis Férussac, 1821 Streptaxis (Odontertemon) glaber (Pfeiffer). Baker, 1925: 39-40 Streptaxis (Streptartemon) glaber Pfeiffer. Venmans, 1963: 53-68, figs. 12-16 Distribution: Brazil; Suriname; Guyana; Venezuela; Isla Margarita; Trinidad; Barba- dos; St. Lucia; Dominica; St. Thomas; St. Croix; Virgin Islands. Status in Barbados: Common; locally abun- dant. Material Collected From: Sites 2, 3, 4, 7, 9, 16, 19, 20, and 22. Additional Material Examined: ANSP 1202 (1 specimen /egit Bland); ANSP 5029 (1 speci- men /egit Bland); ANSP 85445 (11 specimens legit Brown); FLMNH 113853 (Welchman Hall Gully —1 specimen); FLMNH 119894 (Har- rison’s Cave —2 specimens /egit Thompson); MCZ 318941 (near Fresh Water Bay legit Balch). Discussion: Although Férussac's name de- formis has been used by most authors for this streptaxid species, that taxon remains of un- known origin (Baker, 1925). One of the more common species in Barbados, it occurs wher- ever there has been any kind of human dis- turbance. Other than Hughes’ (1750) descrip- tion of snails that may correspond to this species, the first published record is by Smith & Feilden (1891). It is clear that Bland col- lected this species, based on specimens that were deposited in the ANSP collection, but he did not list it in his 1862 publication, possibly because he was unable to identify the speci- mens. Gulella (Huttonella) Bicolor (Hutton, 1834) Pupa bicolor Hutton, 1834: 86, 93 (Mirzapur, India) Ennaea (Huttonella) bicolor (Hutton). Brown, 1903: 269 Distribution: Tropics and subtropics world- wide. Status in Barbados: Unknown. Discussion: This species of Old World origin has been reported as introduced to various Caribbean islands, and there are several lots in the ANSP and FLMNH collections from throughout the West Indies. However, it ap- pears to be fairly uncommon at all localities. Brown (1903) also reported the species from St. Thomas, Dominica, as well as several lo- calities in Barbados (Bridgetown, Belle Plan- tation Wood and St. Philip’s). He is, however, the only worker to report its occurrence on the island. Nevertheless, there is insufficient rea- son to conclude that the species is no longer living on Barbados. Family ZONITIDAE Glyphyalinia barbadensis Chase & Robinson, new name Helix incisa Pfeiffer, 1866: 78 (“Habitat in in- sula Barbados”); 1868: 107, non Helix in- cisa Gmelin, 1791 Hyalina incisa (Pfeiffer). Kobelt, 1880: 284 Vitrea incisa (Pfeiffer). Smith & Feilden, 1891: 249; Brown, 1903: 268 (doubtful occur- rence) LAND SNAILS OF BARBADOS 49 ? Vitrea sp. Henderson, 1919: 96 Retinella (Glyphyalinia) incisa (Pfeiffer). Baker, 1930: 209 (?Retinella (Gly- phyalinia) carolinensis (Cockerell, 1890) Distribution: Endemic to Barbados. Status: Rare. Material Examined: ANSP 997 (3 specimens legit Bland); ANSP 48818 (2 specimens); FLMNH 119896 (Harrison's Cave —4 speci- mens /egit Thompson) Discussion: The identity and even the exis- tence of this species has been confused ever since itwas described by Pfeiffer (1866). It ap- pears not to have been collected by any of the subsequent workers on the Barbados fauna. Kobelt (1880) merely listed it (immediately after the publication of Pfeiffer’s description); Smith & Feilden (1891) and Brown (1903) doubted its occurrence on the island. Smith & Feilden (1891), studying Pfeiffer’s type mate- rial, commented on “the distinct impressed lines of growth which divide the last whorl into numerous segments,” characteristic of the shell of Glyphyalinia. There are two lots matching Pfeiffer's description in the ANSP collection. Both lots were collected from Bar- bados, and one is labelled as having been collected by Bland. Baker (1930) referred to one of the lots as containing potentially mislo- calized specimens of Glyphyalinia carolinen- sis (Cockerell), an eastern North American species. As he also pointed out, the name Helix incisa is pre-occupied, so this taxon lacks a specific name. We therefore propose to provide the name Glyphyalinia barbadensis for this species, one that we consider en- demic to Barbados. Several specimens of the species were collected by Thompson in 1987, confirming the continued survival of this species on the island. Family SYSTROPHIIDAE Miradiscops implicans (Guppy, 1868) Zonites implicans Guppy, 1868b: 440 (Trini- dad) Vitrea implicans (Guppy). Brown, 1903: 268 (doubtful occurrence; attributed to Smith [& Feilden]) Distribution: Venezuela and Trinidad. Status in Barbados: Unknown. Discussion: We cannot confirm that this species was ever collected from Barbados. Although much of Brown’s material (report- edly collected at Porter’s Wood) is deposited at the ANSP and the BMNH, there are no specimens of this species collected by Brown in either of these collections, and no other known records from Barbados. However, the occurrence of the following species, also of this South American family, suggests the pos- sibility that the tiny Miradiscops implicans may have been collected from Barbados. Happiella cf. decolorata (Drouét, 1859) Zonites decolorata Drouét, 1859: 50-51, pl. 1, figs. 3-5 Distribution: Guyana and French Guyana. Status: Rare. Material Collected From: Sites 7 and 13. Additional Material Examined: FLMNH 119897 (Harrison’s Cave —4 specimens legit Thompson). Discussion: Examination of specimens col- lected at two localities during this study shows them to be virtually indistinguishable in terms of shell morphology from specimens of Hap- piella cf. decolorata (Drouét, 1859), a species known from Guyana and French Guiana. The generic placement of this species follows that of Ramírez (1993). Family MEGALOBULIMIDAE Megalobulimus oblongus (Múller, 1774) Helix oblongus Múller, 1774: 86 Bulimus oblongus (Muller). Bland, 1862: 351; Kobelt, 1880: 284; Rush, 1891: 65, 69; Smith, 1895: 302, 306 Bulimus (Borus) oblongus (Muller). Smith & Feilden, 1891: 251-252 Strophocheilus (Borus) oblongus Müller. Brown, 1903: 268; Henderson, 1919: 94-95 Distribution: South America; Trinidad; To- bago; Grenada; St Vincent; Barbados. Status in Barbados: Uncommon. Material Collected From: Sites 2, 4, 5, 7, 14, and 16. Fragmentary specimen (observed, not collected) at site 19. Additional Material Examined: ANSP 83178 (1 specimen /egit Brown); ANSP 85470 (3 specimens /egit Brown); RM 13457 (2 speci- 50 CHASE 8 ROBINSON mens /egit Rawson ex Carpenter); MCZ 232161 (Lancaster Plantation, St. James /egit Gooding); MCZ 230453 (Airy Hill, St. John — 1 specimen /egit Humes); MCZ 90359 (6 speci- mens /egit Junious ex Bland); MCZ 50505 (Bridgetown — 2 speciments /egit Walker). Discussion: Megalobulimus oblongus 1$ widely distributed throughout the island, al- though it was considerably more common in the past. Rush (1891) remarked on its abun- dance in Bridgetown, and Henderson (1919) reported the species as living “abundantly all over the island especially in gardens.” Bland (1862) believed the species was introduced to Barbados from St. Vincent by Rev. J. Parkin- son, although the purpose for the introduction is unknown. Family SAGDIDAE Lacteoluna selenina (Gould, 1848) Helix selenina Gould, 1848: 38 (Georgia and Florida) Helix vortex Pfeiffer. Bland, 1862: 351; Pfeif- fer, 1876: 153 (reported from Barbados), non Linné, 1758 Helix (Microphysa) vortex (Pfeiffer). Smith & Feilden, 1891: 251 Microphysa vortex (Pfeiffer). Kobelt, 1880: 284 Helix subaquila Shuttleworth. Rush, 1891: 68, non Shuttleworth, 1854 Thysanophora vortex (Pfeiffer). Brown, 1903: 268 ? Thysanophora sp. Henderson, 1919: 96 Lacteoluna selenina barbadensis Pilsbry, 1930: 244 (Barbados); Baker, 1963: 242 Distribution: Bermuda; Florida and the West Indies. Status in Barbados: Rare. Material Collected From: Site 9. Additional Material Examined: ANSP 8039 (lectotype of L. s. barbadensis Pilsbry, 1930, designated by Baker, 1963); ANSP 28317 (11 paralectotypes of L. s. barbadensis Pilsbry); ANSP 85469 (2 specimens /egit Brown); MCZ 905 (Blowers —3 specimens /egit Kugler). Discussion: This synanthropic species, widely distributed throughout the Caribbean Basin, appears not to be common in Barba- dos. Pilsbry (1930) considered the Barbados populations as a distinct subspecies because they are a little larger than is typical for the species. Smith & Feilden (1891) reported 2 specimens, Brown deposited 2 specimens in the ANSP collection, and 2 specimens were collected during this study. Lacteoluna (Aerotrochus) turbiniformis (Pfeiffer, 1839) Helix turbiniformis Pfeiffer, 1839: 350 (Cuba) Helix (Microphysa) turbiniformis Pfeiffer. Smith & Feilden, 1891: 251 (uncon- firmed) Thysanophora turbiniformis (Pfeiffer). Brown, 1903: 266, 268 (doubtful occurrence; at- tributed to earlier workers) Distribution: Cuba and Jamaica. Status in Barbados: Dubious record. Discussion: Smith & Feilden (1891) reported a single specimen of this species, reputedly from Barbados, in the Cuming collection (in the British Museum). However, they also noted that the collection was “somewhat no- torious for errors of locality” (Smith & Feilden, 1891: 248), and they could not confirm the occurrence of the species based on Feilden’s collections from Barbados. No other workers reported the species, and Brown (1903) con- cluded that its occurrence was doubtful. We concur with earlier workers that this Greater Antillean species was erroneously reported from Barbados. Family CAMAENIDAE Pleurodonte isabella (Férussac, 1822) Helix Isabella Férussac, 1821: 32, (“Les An- tiles, Cayenne”) (nomen nudum); Bland, 1862: 351, table 2 Helicogena isabella Férussac, 1822a: pl. 47, fig. 2 Helix Barbadensis Lamarck, 1822: 78-79 (“la Barbade”) Carocolla Barbadensis Guilding, 1828b: 167 (“sub lapidibus Barbadensibus”) Helix (Dentellaria) Barbadensis Lamarck. Beck, 1837: 35 Helix barbadensis (Carocolla) Guilding. Pfeif- fer, 1847: 310 Helix dentiens Férussac. Deshayes, 1850: 147-148, pl. 47, fig. 2 only; Pfeiffer, LAND SNAILS OF BARBADOS 51 1853: 213 (partim non Helix dentiens Férussac) Dentellaria Isabella (Férussac). Kobelt, 1880: 284 Helix (Dentellaria) isabella Férussac. Pilsbry, 1889: 85-86, pl. 24, figs. 42, 43; E. A. Smith & Feilden, 1891: 250 Pleurodonte isabella Férussac. Brown, 1903: 268 Pleurodonte (Caprinus) isabella (Férussac). Henderson, 1919: 95 Distribution: Endemic to Barbados. Status: Locally abundant. Material Collected From: Sites 2, 3, 4, 5, 7, 8, ЗЫ: 16, 172 20F 21 Fandı22 Additional Material Examined: ANSP 987 (3 specimens), ANSP 988 (4 specimens), ANSP 989 (1 specimen), ANSP 990 (4 specimens), ANSP 991 (3 specimens), ANSP 992 (3 spec- imens), ANSP 993 (2 specimens), ANSP 994 (1 specimen), ANSP 995 (2 specimens), ANSP 32584 (8 specimens), all legit Bland; ANSP 85443 (8 specimens /egit Brown); ANSP 85444 (2 specimens /egit Brown); RM 14301 (9 specimens legit Rawson ex Carpen- ter); FMNH 146396 (Wentwood Gully — 1 specimen); FMNH 147023 (Codrington Col- lege —4 specimens); FMNH 172045 (Speight- stown —6 specimens); MCZ 251322 (Farley Hill, St. Peter—15 specimens legit Ray 8 Allen); MCZ 258049 (4 miles east of Hole- town, St. James-4 specimens /egit Scheafer); MCZ 136004 (Christ Church —1 specimen legit Bland); MCZ 251320 (cave near Hillaby, St. Thomas —9 specimens legit Ray & Allen); MCZ 107935 (Blowers — 10 specimens /egit Kugler); MCZ 108727 (St. Joseph’s —1 specimen /egit Cockerell); MCZ 90325 (1 specimen ex Bland). Discussion: Pleurodonte isabella is a ubiqui- tous Barbados endemic that appears to be relatively unaffected by human activity on the island, being extremely common in suburban gardens as well as relatively unspoiled forest areas. Pleurodonte dentiens (Férussac, 1822) Helix (Helicodonta) dentiens Férussac, 1821: 33 (“La Guadeloupe, la Martinique, Saint- Domingue, les forêts de Cayenne et de la Guyanne”) (nomen nudum) Helix dentiens Férussac, 1822b: pl. 48, fig. 2 Helix dentiens Ferussac. Deshayes, 1850: 147-148; Pfeiffer, 1853: 213 (partim) Pleurodonte dentiens (Férussac). Brown, 1903: 268 (unconfirmed) Distribution: Guadeloupe, Martinique. Dominica, and Status in Barbados: Erroneous record. Discussion: Although Brown (1903) reported that Pleurodonte dentiens had been collected “by a Mr. E. W. Williams” in Pine Wood Estate, St. Michael's, he could not personally confirm the occurrence of this species. Apparently the confusion was created when both Deshayes (1850) and Pfeiffer (1853) placed Helix ¡s- abella as a junior synonym of Helix dentiens, and Brown was unaware of this taxonomic error. Polydontes perplexa (Férussac, 1832) Helix (Helicodonta) perplexa Férussac, 1832: pl. 56A, fig. 1 Helix (Dentellaria) perplexa Férussac. Smith 8 Feilden, 1891: 249 Pleurodonte perplexa (Férussac). Brown, 1903: 268 (doubtful occurrence; attrib- uted to Smith [8 Feilden]) Distribution: Grenada and the Grenadines. Status in Barbados: Dubious record. Material Examined: BMNH 70.10.12.7 (4 specimens /egit Rawson). Discussion: Smith & Feilden (1891) referred to specimens of Helix perplexa Férussac, as being “said to have come from that locality” [Barbados] that were collected by Sir Rawson and deposited at the British Museum (Natural History). Smith later (1895) listed this species from Grenada and the Grenadines, but not from Barbados. We have examined these specimens (BMNH 70.10.12.7), and although the identification can be confirmed, it is un- likely that they were collected from Barbados. Brown (1903) also was unable to confirm the occurrence of this species on the island. Zachrysia provisoria (Pfeiffer, 1858) Helix provisoria Pfeiffer, 1858: 39-40 (Man- zanillo, Cauto, and Guisa, Cuba) 52 CHASE & ROBINSON Distribution: Originally from Cuba; estab- lished in southern Florida, the Bahamas, St. Croix, and Barbados. Status in Barbados: Locally abundant. Material Collected From: Sites 1, 2, 3, 4, 13, and 15. Discussion: Although this large Cuban species has well established populations in southern Florida, as well as the Bahamas (Pilsbry, 1928), it has not been reported until now from anywhere in the Lesser Antilles. Nevertheless, this recently introduced arrival is particularly common in lowland parts of the island, especially along the East coast. In some areas, especially in gardens, it is the most conspicuous snail species. It is likely to have been introduced via the horticultural trade, possibly from Florida. Family BRADYBAENIDAE Bradybaena similaris (Rang, 1831) Helix similaris Férussac, 1821: 43 (“Timor”) (nomen nudum) Hélice (Helicelle) similaris “Férussac” Rang, 1831: 15 (“Bourbon . . . l’île de Cuba; ... Rio-Janeiro et particulièrement au jardin de Saint-Christophe . . . Brésil.”); Bland, 1855: 153; 1862: 351, table 2 Fruticicola similaris “Férussac.” Kobelt, 1880: 284 Helix (Fruticicola) similaris “Férussac.” Smith & Feilden, 1891: 250-251 Helix (Dorcasia) similaris “Férussac.” Rush, 1891: 69 Eulota similaris “Férussac.” Brown, 1903: 268; Henderson, 1919: 96 Distribution: Originally eastern Asia; estab- lished throughout the tropics and subtropics. Status in Barbados: Locally common. Material Collected From: Sites 3, 23 and 24. Additional Material Examined: ANSP 998 (4 specimens /egit Bland); ANSP 85448 (4 spec- imens legit Brown); ANSP 62063 (2 speci- mens /egit Rush); MCZ 136009 (5 specimens ex Blana). Discussion: First reported from Barbados by Bland (1855), Smith 8 Feilden (1891: 250) later reported this species as being “the com- monest Helix in the island,” presumably more so than Pleurodonte isabella, which they also referred to as a Helix. They also noted that it was “abundant on the lowlands as well as on the high ground of Scotland District. . . .” Today, Bradybaena similaris appears to be much more restricted in distribution. In our survey, it was found in only three localities, both in St. James Parish, where it is common but considerably less so than Zachrysia provi- soria and Pleurodonte isabella. DISCUSSION We have described significant discrepan- cies in the faunal lists obtained by seven groups of workers in Barbados, including our- selves. The variability cannot easily be attrib- uted to simple incompetence, because all the groups comprised, or at least included, per- sons with considerable experience as collec- tors and malacologists. While the results of our study clarify the current conservation sta- tus of land snails on Barbados, we believe they also have important implications for con- servation efforts generally. The reported faunal lists for Barbados sep- arate into two groups according to the length of the list, which presumably reflects the in- tensity of the collection effort (Table 1). In one group, the lists of Carpenter, Rush and Hen- derson are very likely incomplete. In the sec- ond group, comprising Bland, Smith and Feilden, Brown, and ourselves, the total num- bers of confirmed species range from 19 to 23. The absence of any evident historical trend in these totals should not be taken to imply an absence of faunal change over the intervening 137 years. On the contrary, faunal change is indicated by our analysis in Table 2, which shows consistent disagreement among the workers with respect to the content of the lists. For example, the earliest worker, Bland, reported five confirmed species that were not found by us, and conversely, we found five species not reported by Bland. Similarly, Smith & Feilden reported three confirmed species not found by us, and we found six species not reported by Smith & Feilden. When Smith & Feilden’s list is compared with Brown's, there is one “different” species re- ported by Smith & Feilden and five “different” species reported by Brown. Given that the foregoing account is based exclusively on LAND SNAILS OF BARBADOS 53 taxa for which the location in Barbados has been confirmed by our examination of institu- tional collections, or by other workers, the pattern of inconsistency is likely to reflect a combination of sampling error and actual fluc- tuations in the fauna. To know the exact size of the current fauna would require a more thorough field survey than we were able to conduct. All together, we confirmed 30 species from our own survey plus those of earlier authors (Table 1) In addi- tion, we confirmed Pupisoma dioscoricola from the FLMNH collection, to give a total of 31 confirmed species present on the island at some time in the period 1862 to the present. Beyond these confirmed species, Miradis- cops implicans and Gulella bicolor are also likely to have been found on the island, al- though we did not examine any specimens, and some small, inconspicuous species may have escaped the notice of all collectors. On the other hand, three endemic species (Bu- limus fuscus, Lucidella barbadensis and Pseudopineria barbadensis) are probably ex- tinct, and one introducted species (Lucidella plicatula) is possibly extirpated. In summary, the total number of extant species is about 30, or slightly less. While we confirm 31 species for Barbados in the period 1862 to the present, a total of 46 species were listed by the six authors whose works have been analyzed here. We also found 12 additional species reported by other workers scattered throughout the malacologi- cal literature. The disparity between the num- ber of confirmed species (31) and the total number of reported species (58) can be at- tributed to a combination of synonyms and er- roneous reports. Brown's (1903) faunal list has special sig- nificance because it is presumably the basis for a widely circulated estimate of the number of species present on Barbados. We refer to a report (Groombridge, 1992) complied by the World Conservation Monitoring Centre in col- laboration with a number of highly respected international conservation organisations. This report contains a list (p. 151, table 14.3) of the total number of land snails on various islands. Data in the list were compiled by the Species Survival Commission Mollusc Specialist Group of the International Union for Conser- vation of Nature and Natural Resources (IUCN/SSC), but without specific references. Barbados is said to have 37 species, which is the exact, and unique, number reported by Brown (1903). In light of the special status af- forded to Brown's total, and noting that we have been able to confirm the validity of only 23 of the 37 “species” from his list (Table 1), a detailed analysis of Brown's list is warranted. In addition to the 23 confirmed species, two species are unconfirmed but likely to occur in Barbados, five species are correctly identified but of dubious locality, and five species were listed by Brown only by attribution to others (Table 3). By this reckoning, the total of con- firmed and unconfirmed species from Brown's list is 35. The two remaining taxa from Brown's list are redundant synonyms: Opeas goodalli and Opeas ascendens are both Opeas hannense; Cecilioides gundlachi and Opeas gundlachi are both Cecilioides aperta. Brown introduced further confusion by la- belling as Cecilioides aperta specimens that are in fact Cecilioides consobrinus minutis- sima. No doubt the reason that Groombridge (1992) lists the land snail fauna of 86 islands is that islands have been well studied by malacologists (e.g., Peake, 1981; Solem, 1984; Cowie, 1997). On the face of it, the data set is a good basis for monitoring international conservation efforts. However, given the like- lihood that islands other than Barbados also have erroneous entries, the information con- tained therein, particularly by older workers, must be viewed with caution. Additionally, the notoriously disputed taxonomy of pulmonate gastropods can create major problems for modern workers. In our taxonomical section, in which 38 species are treated, 136 variant names are listed, covering synonyms, mis- identifications and variant genus/species combinations, but excluding different taxon authorships (often incorrect) and citations at the genus level only. We think it appropriate to advise that serious conservation work on many of these islands must be preceded by careful checking of the historical records. A similar laborious effort will be required for many non-island localities. Our survey results can be compared with those of earlier workers to assess the conser- vation status of the Barbadian land snails, al- beit with the reservations implicit in the com- ments above. It is evident, from all records, that species diversity on Barbados (430 km’) is low relative to other small Caribbean is- lands, for example, Saba (13 km”, 14 species; Clench, 1970) and St. Martin (98.5 km?, 39 species; Coomans, 1967). This is likely a re- 54 CHASE 8 ROBINSON sult of several factors, but principally the ab- sence of mountains on Barbados, the large distance of Barbados from neighboring is- lands, and the fact that Barbados lies to the east of other islands, hence upwind and up- current. In our Surveys, we found six endemic species, five species native to the Lesser An- tiles and/or South America, three pan- Caribbean species and seven pan-tropical species. Conservation interest naturally fo- cuses on the endemic species, of which three were not found by us, including one, Bulimus fuscus, that is almost certainly extinct. On the other hand, we located specimens of Gly- phyalinia barbadensis, which had been con- sidered as an endemic based on Pfeiffer’s original description of Helix incisa (1866), but which had not been subsequently confirmed for the Barbados locality. Even if Lucidella barbadensis and Pseudopineria barbadensis are eventually found, it is clear that the fauna is dominated by introduced taxa. Of these, several synanthropic species are particularly abundant, namely Bulimulus guadalupensis, Allopeas gracile, Allopeas micra, Beckianum beckianum, Leptinaria lamellata, Zachrysia provisoria and Streptaxis glaber. The synan- thropes represent a group of invasive species that are spreading by human transportation, hence referred to as “traveling species” (Robinson, 1999). While the overall diversity of land snails ap- pears to be approximately as great today as it was 150 years ago, its composition has evolved over time. Change will continue as new species are introduced from elsewhere (Cowie, 1998, Robinson, 1999), and tropical faunas worldwide become increasingly ho- mogenized. The synanthropic species, whether of Antillean or pan-tropical origin, are clearly tne most abundant throughout Barba- dos, as little if any of the island remains in a truly undisturbed state. Whether the synan- thropes have actually displaced native species, or have simply occupied increasingly available habitats created by man, needs to be investigated. Of those endemic species that have survived, only those that are rela- tively catholic in their ecological requirements, or those whose natural habitat approximates an anthropochorous environment, seem to be maintaining stable populations. The remain- der seem destined to become extinct, if they have not already become so. In any case, it is clear from our study that any effort to under- stand the snail fauna of Barbados, or any other locality, with a view to identifying and protecting its indigenous elements, must carefully evaluate the evidence of historical trends. 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Studies on the Fauna of Curacao and Other Caribbean Is- lands, 1: 59-108. WAGNER, A. J., 1907-1911, Die Familie der He- licinidae. Systematisches Conchylien-Cabinet von Martini & Chemnitz, (n. s.)1(18:2): 1-391, pls. 1-170. WAREBORN, I., 1992, Changes in the land mollusc fauna and soil chemistry in an inland district in southern Sweden. Ecography, 15: 62-69. WILLING, M. J., 1998, The conchological society and molluscan conservation: Past, present and future. Pp. 139-145, In 1. J. KILLEEN, M. В. SEDDON. & A. M. HOLMES, Molluscan conservation: a strat- egy for the 21st century, Journal of Conchology, Special Publication, Revised ms. accepted 19 July 2000 APPENDIX The following specimens collected during this study have been deposited as vouchers in the Department of Malacology at the Acad- emy of Natural Sciences, Philadelphia. Helicina fasciata substriata Gray, 1824 — ANSP 399360 8 399361 Truncatella barbadensis Pfeiffer, 1857 — ANSP 401921 8 401922 Succinea barbadensis Guilding, 1828 — ANSP 399354 Gastrocopta barbadensis (Pfeiffer, 1853)— ANSP 401923 Bulimulus guadalupensis (Bruguiére, 1789)— ANSP 399359 Orthalicus maracaibensis subpulchella (Pils- bry, 1889) -ANSP 399357 Brachypodella costata (Guilding, 1828)— ANSP 399362 Allopeas gracile (Hutton, 1834)—ANSP 401924 Allopeas micra (d'Orbigny, 1835)-ANSP 401925 Beckianum beckianum (Pfeiffer, 1846)— ANSP 401926 Leptinaria lamellata (Potiez 8 Michaud, 1835)-ANSP 401927 Luntia insignis Smith, 1898 —ANSP 401928 Opeas hannense (Rang, 1831)-ANSP 401929 Subulina octona (Bruguiére, 1792)-ANSP 399364 Streptaxis glaber (Pfeiffer, 1849)-ANSP 399355 Happiella cf. decolorata (Drouét, 1859)— ANSP 401930 Megalobulimus oblongus (Müller, 1774)— ANSP 399356 Lacteoluna selenina (Gould, 1848)-ANSP 401931 Pleurodonte isabella ANSP 399358 Zachrysia provisoria (Pfeiffer, 1858)— ANSP 399363 & A18831 Bradybaena similaris (Rang, 1831)-ANSP 401932 Synoptic collections have been deposited at the Bellairs Research Institute of McGill University, Holetown, Barbados, and the De- partment of Biological and Chemical Sci- ences, University of West Indies, Cave Hill campus, St. Michael, Barbados. (Férussac, 1822)— NOTE ADDED IN PROOFS Subsequent to completion of the manu- script we received one specimen of Achatina са Bowdich, 1833, from Barbados. Its in- troduction probably occurred within the previ- ous year, but its distribution is yet to be deter- mined. MALACOLOGIA, 2001, 43(1-2): 59-85 REPRODUCTION, DURATION OF EMBRYOGENESIS, EGG CAPSULES AND PROTOCONCHS OF GASTROPODS OF THE FAMILY BAICALIIDAE (CAENOGASTROPODA) ENDEMIC TO LAKE BAIKAL T. Sitnikova', P. Ropstorf* & Е. Riedel? ABSTRACT Data on the reproduction and duration of embryogenesis have been obtained for the first time for the Lake Baikal endemic gastropods of the family Baicaliidae. Their reproduction occurs throughout the year, but with peaks in spring and autumn. Embryogenesis duration supposedly depends on the near-bottom water temperature. Hatching appears to be timed to coincide with the periods of increased food supplies. Egg capsule and protoconch morphology has been ex- amined in several species, and four types of capsules are described. Shells of the Baicaliidae are orthostrophic, and only in Liobaicalia stiedae does the protoconch axis differ significantly from the teleoconch axis, which was previously mistaken for heterostrophy. Three groups were distinguished on the basis of a cluster analysis of size and number of protoconch whorls. These clusters do not fully coincide with the recent taxonomy of the Baicaliidae based on shell mor- phology and the female reproductive system. Some species assigned to the same genus or sub- genus differ either in protoconch size or egg capsule morphology. Key words: ancient lakes, Baikal, rissooidean gastropods, egg capsules, embryogenesis, pro- toconchs. INTRODUCTION More than 100 species of Gastropoda are known to inhabit Lake Baikal; of these, 40 are assigned to the family Baicaliidae (Sitnikova, 1994) following the classification of Starobogatov & Sitnikova (1983), or to the subfamily Baicaliinae of the family Hydrobi- idae according to Ponder & Waren (1988). Baicaliids are one of the least studies groups of endemic molluscs in Lake Baikal. Never before has their embryonic shell been closely examined, though at least one baicaliid— Li- obaicalia stiedae (Dybowski, 1875)—was re- ported to possess a heterostrophic shell by Dybowski (1875), and this information is in- cluded in various reference books on gas- tropods. Egg capsule morphology data are scarce: only three species (without precise identification) have been previously described (Sitnikova, 1991a). There is information (Gavrilov, 1953) on the reproduction of only one species, Maackia (Eubaicalia) herderiana (Gerstfeldt, 1859). Our initial goal was to study egg capsule and protoconch morphol- ogy, but in the process we obtained additional data relevant to the duration of reproduction and embryogenesis in several species. These data are included in this paper. MATERIALS AND METHODS Living gastropods and egg masses were collected by SCUBA divers or by means of a dredge or grab in 1995-1997 (Table 1, Fig. 1). The egg capsules were kept in 10 ml glass vials until hatching in a refrigerator (6°C). Ma- terial of previous expeditions, from other lo- calities, stored at the Limnological Institute, was used for comparison. Some adult gastropods were brought alive to the Institute of Ecological Toxicology in Baikalsk, where a steady stream of Lake Baikal water (from a depth of 40 m) with a constant temperature of 6°C was maintained. Each species was cultivated separately in a 20 | glass vessel. In order to obtain egg cap- sules from a single species, all egg clutches laid on the shells prior to the experiment were removed. The clutches were kept in aquaria from July 1995 until May 1996 and were checked daily until the end of September, and then weekly until November. ¿Limnological Institute, P.O. Box 4199, Irkutsk, 664033, Russia; sitOlin.irk.ru Institute of Palaeontology, Freie Universitat Berlin, Germany; palaeont @ zedat.fu-berlin.de 59 60 SITNIKOVA, ROPSTORF & RIEDEL TABLE 1. List of investigated species and sites of their collection. n Site 1 Baklany Rock next to Pestchanaya, sand, 5-10 m, 07.16.1995 2 Bolshie Koty, sand, 9-10 m, 16.07.1995 3 Bolshie Koty, Zhilische canyon, rock, 3-15 m, 27.07.1995, 14.12.1997 4 Bolshie Koty, sand, 15-18 m, 27.07.1995 5 Kultuk, Shaman Cape, silty sand, 20 m, 27.08.1996 6 Maloe More Strait, Ushun Cape, sand, 10-12 m, 13.10.1995 7 Maloe More Strait, Olkhon Gate, grey sand, 27-31 m, 13.10.1995 8 Maloe More Strait, Ushun Cape, stones, 3 and 14 m, 13.10.1995 9 Maloe More Strait, Shaman Rock, stones, 5 m, 14.09.1996 10 Chivyrkui Bay, sand, 20-28 m, 20-25 m, 16.10.1995 11 Chivyrkui Bay, Kyltygei Island, stones, 1.5-2 m, 08.10.1997 12 Svyatoi Nos Peninsula, stones, 6-8.5 m, 15.10.1995 13 Boguchany, stones, 3 m, 18.10.1995 14 Muzhinai, silty sand, 20-64 m, 20.10.1995 15 Barguzin Bay, sand, 25 m, 14.10.1995 Species Parabaikalia oviformis (Dybowski, 1875); P. elata elata (Dybowski, 1875); Korotnewia semenkewitschi (Lindholm, 1909) Parabaikalia florii florii (Dybowski, 1875); P. elata elata (W. Dybowski, 1875) Baicalia turriformis Lindholm, 1909; Maackia (Eubaicalia) bythiniopsis (Lindholm, 1909); M. (E.) herderiana (Gerstfeldt, 1859); M. (Maackia) costata (Dybowski, 1875) Liobaicalia stiedae (Dybowski, 1875); Pseudobaikalia (Pseudobaikalia) contabulata (Dybowski, 1875); P (P) zachwatkini Kozhov, 1936 Liobaicalia stiedae (W. Dybowski, 1875); Baicalia cari- nata (W. Dybowski, 1875); Pseudobaikalia (P.) con- tabulata (Dybowski, 1875); P. (Microbacalia) pulla pulla (Dybowski, 1875) Parabaikalia florii kobeltiana Lindholm, 1909; P. elata elata (Dybowski, 1875) Teratobaikalia (Baicaliella) nana (Lindholm, 1909), T. (B). nana f. clandestina Beckman & Starobogatov, 1975; Parabaikalia florii kobeltiana Lindholm, 1909 Maackia (Eubaicalia) bythiniopsis (Lindholm, 1909); M. (E.) variesculpta (Lindholm, 1909); Teratobaikalia (Teratobaikalia) macrostoma Lindholm, 1909 Teratobaikalia ( Teratobaikalia) macrostoma Lindholm, 1909 Parabaikalia elata dubiosa (Kozhov, 1936); Pseudo- baikalia (Pseudobaikalia) contabulata (Dybowski, 1875); Teratobaikalia (Baikaliella) nana f. clandestina Beckman & Starobogatov, 1975; T. (B.) nana f. pro- ducta Beckman & Starobogatov, 1975; T. (Trichio- baikalia) duthiersii (Dybowski, 1875); Korotnewia semenkewitschi (Lindholm, 1909); Baicalia carinata (Dybowski, 1875); B. carinatocostata (Dybowski, 1875); Godlewskia pulchella (W. Dybowski, 1875) Teratobaikalia ( Teratobaikalia) macrostoma Lindholm, 1909 Maackia (Eubaicalia) variesculpta (Lindholm, 1909); M. (E.) bythiniopsis Lindholm, 1909 Maackia (Eubaicalia) variesculpta (Lindholm, 1909); Baicalia dybowskiana Lindholm, 1909M. (E.) bythin- iopsis Lindholm, 1909; Teratobaikalia ( Trichiobaikalia) ciliata (W. Dybowski, 1875); Baicalia carinata (W. Dybowski, 1875); P. oviformis (W. Dybowski, 1875) Parabaikalia elata elata (W. Dybowski, 1875); Teratobaikalia (Baikaliella) nana clandestina Beckman & Starobogatov, 1975; T. (B.) nana humerosa Beckman & Starobogatov, 1975; Pseudobaikalia (Microbaicalia) pulla tenuicosta (Lindholm, 1909) Hatched snails and juveniles with intact protoconchs were fixed in 70% ethanol. Egg capsules unused in the experiments were fixed in 4% formaldehyde or 2.5% glutaralde- hyde. The shells of hatched embryos and older ju- venile gastropods were examined and mea- sured using a Cambridge 360 SEM (Scanning Electron Microscope) in Berlin. Calcification of the embryonic shell was studied with a Zeiss Axiophot microscope using polarized light. The term “protoconch” can be applied here to the embryonic shell, because baicaliids have no free larvae and hence no larval shells. In REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 61 Svyatoi Nos Peninsula Shaman Rock Maloe More Ushun Cape — À Pestchanaya Bolshie Angara ae, River IN Kultuk ask Kyltygei FE Chivyrkui Ba Barguzin Bay Selenga River FIG. 1. Map of Lake Baikal with indication of collection sites. addition to the description of their sculpture, four measurements are given: number of whorls, maximum diameter, maximum width of the non-spiral of the primordial shell (Fig. 2), and the height from the apex to the end of the aperture. Calculation of the temperature sum (de- gree-days): The data on day-to-day variations of water temperature in near-bottom waters of the littoral zone of Lake Baikal are lacking. Ac- cording to Prof. Shimaraev (Limnological In- stitute, Irkutsk, personal communication), an expert on the thermal regime of the lake, it is possible to estimate approximately the tem- 62 SITNIKOVA, RÓPSTORF & RIEDEL N FIG. 2. Apical view of a protoconch with explana- tions of measurements. | = initial point (axis of counting whorls), MD = maximal diameter, N = non- spiral (from Riedel, 1993). perature sum (degree-days) by using monthly averages of water temperatures basing on data for 1897-1902 for the Goloustnoye re- gion at depths of 5, 10, and 20 m. (Voznesen- sky, 1908) and for 1911-1916 for Peschanaya Bay at 5, 10, 20 m depths (Verbolov et al., 1965). These data are presented in Figure 3. We chose the temperature regimes о the 5, 10 and 20 m depths, because the gastropods for which the egg-laying time and duration of embryonic development were investigated occur in large quantities at depths above 20 m. The time of embryonic development of the gastropods is given (in days), and ambient water temperature in the laboratory or that es- timated in the lake (in °C), which allows a cal- culation of the duration of embryogenesis in degree-days (or temperature sum), that is, av- erage temperature in °C multiplied by the number of days. Statistical processing (T-test, Cluster and Factor analysis) of data was performed using the Statistica 4.3 software package. Results were analysed and reported by T. Y. Sitnikova. Most adult specimens are stored at the Lim- nological Institute of Siberian Branch of the Russian Academy of Sciences in Irkutsk. The SEM-preparations of protoconchs and em- bryos were deposited at the Institute of Palaeontology of the Free University of Berlin. RESULTS Duration of Embryogenesis Data on the reproduction of 15 species of baicaliids were obtained. Observations on em- bryonic development in culture have shown that when the egg capsules are laid, they con- tain a round, yellow embryo with no deter- mined structure of about 0.3 mm in diameter. After 1-3 mo, the embryo begins to develop a foot, head and visceral mass. After another 2-3 mo, the visceral mass is covered by an or- ganic shell, showing a pit (Fig. 11B). Calcifica- tion starts from the apical part of the shell. The margin of the mantle is extremely broad in all species examined (Fig. 7A). When the shell has reached 0.5-0.75 whorls, the operculum has formed, but at that time the embryo is un- able to retract into the shell. A fully developed young snail occupies the whole lumen of its capsule. The snails always hatch through a hole at the top of the capsule. Shell growth stops for a while before the embryos leave their capsules, despite their in- tense consumption of intracapsular sub- stance and excretion of faecal pellets. The duration of embryonic development was observed in eight species (Table 2). They laid clutches in the laboratory in autumn (Sep- tember-November). Under a constant tem- perature of 6°C development lasted 6-9 mo, but the actual temperature of near-bottom water at 10 m depth rises as high as 10-12°C in summer (Kozhov, 1963). Thus, the duration of the in vivo development of embryos ap- pears to be different. To calculate the duration of embryogenesis in baicaliids in their natural conditions, we have selected four species: Maackia herderi- ana, Baicalia carinata, Parabaikalia florii, and Liobaicalia stiedae. Conditions were different for each. The first species occurs in great quantities at 5 m depth, the second and third at 5-10 m and deeper, the fourth at 15-20 m and deeper. Maackia herderiana: in laboratory culture, embryogenesis was equivalent to 1374 de- gree-days, egg capsules were laid in Septem- ber, and the young hatched in April. But the temperature sum in the lake at 5 m depth dur- ing the same period (from September to April) was calculated to be only 811.8 degree-days; that is, under natural conditions, embryoge- nies should last longer. The young were pre- dicted to hatch in mid-August. For females laying capsules in mid-June, embryogenesis at 5 m depth may last about 7.5 mo, and the young will hatch in February (Table 3). This calculated duration of embryogenesis is con- sistent with what we have observed in nature: M. herderiana eggs collected at 5 m depth in October contained some mid-mature em- bryos (Table 4). Baicalia carinata: in laboratory culture, em- bryogenesis was 1290 degree-days and REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 63 IV V VI VII month VIII IX хх "XII —— 5 m --D--10m —X—- 20m FIG. 3. Temperature sum (degree-days) of water in different depths per month, calculated after Voznesen- sky (1908) and Verbolov et al. (1965). TABLE 2. Data and duration of the embryogenesis in culture some of baicaliids collected from different regions of the Lake. Species Parabaikalia florii P. oviformis P elata Maackia herderiana M. variesculpta Baicalia carinata Pseudobaikalia zachwatkini Liobaicalia stiedae Site Maloe More strait, Olkhon Gate, Ushun Cape, 10- 12 m, sand, 13.10.1995 Southern Baikal, Baklany Rock 5-10 m, 16.07.1995 Southern Baikal, Baklany Rock, 5-10 m, 16.07.1995 Southern Baikal, Bolshie Koty, Zhilische canyon, rock, 3-15 m, 27.07.1995 Boguchany, stones, 3 m, 18.10.1995 Kultuk, Shaman Cape, silty sand, 20 m, 27.08.1996 Kultuk, Shaman Cape, silty sand, 20 m, 27.08.1996 Kultuk, Shaman Cape, 20 m, sand, 27.08.1996 Data of egg capsule laid in culture (number of capsules) 01.11.1995 (n = 4) 15.08-26.09.1995 (mail) 15.08.-09.09.1995 (n = 3) 23.08-29.09.1995 (n = 3) 31.10.1995-10.10.1996 (n = 3) 09.10.1995 (n = 1) 01.09-03.10.1996 (n = 6) 03.09.1996 (n = 4) Duration of embryogenesis in culture, in average days (range) 240 (238-245) 216 (170-253) 217 (192-253) 229 (207-250) 344 (n = 1) 215 190 (186-194) (n = 2) 180 (174-186) lasted from October to April. Summed tem- perature in the lake at 10 m depth from Octo- ber to April inclusive was calculated to be 500 degree-days. Under natural conditions, the embryogenesis of B. carinata in capsules laid in October has been predicted to last 11.5 mo at 10 m depth (the young hatching in Septem- ber) and 11 mo at 20 m depth. For eggs laid in mid-June, embryogenesis may last 9.5 mo for 10 m depth (the young will hatch in March), and 11.5 mo at 20 m depth (the snails will hatch in about May). (The summed yearly 64 SITNIKOVA, RÓPSTORF & RIEDEL TABLE 3. Duration of the embryogenesis of some species in culture. Species Duration (degree* days) of embryogenesis in culture (in days) Maackia herderiana Baicalia carinata Liobaicalia stiedae Parabaikalia florii 1374 (229) 1290 (215) 1080 (180) 1440 (240) and time egg capsules were laid Predicted depth 5 m, September 5 m, mid-June 10 m, October 10 m, mid-June 20 m, mid-October 20 m, June 20 m, September 5 m, November 5 m, June 10 m, November 10 m, June 20 m, November 20 m, June Predicted time of embryogenesis (in month) TABLE 4. Data on the development of embryos collected from their natural habitat. Species Maackia bythin- lopsis M. herderiana M. variesculpta Parabaikalia florii florii P. elata elata P. oviformis Baicalia carinata B. dybowskiana Godlewskia pul- chella Teratobaikalia macrostoma T. ciliata Site Northern and Middle Baikal Southern Baikal different regions Northern Baikal Maloe more strait (Middle Baikal) Southern Baikal Southern Baikal Northern Baikal Northern Baikal Northern Baikal Date of collected egg capsule October 1995 December 1997 October 1995 and December 1997 mid-October 1995 July 1995 July 1995 July 1995 August 1995 October 1995 October 1995 October 1995 October 1997 October 1995 Egg capsulae Stage of embryos Substrate number developing stones 27 various: from the first stage of cleavage of a fairly developed embryo stones 15 embryos with a devel- oped shell stones 35 embryos possessing a foot, head and vis- ceral mass stones 57 shell-covered embryos large shells = 50 shell-covered embryos (own and Benedictia) the same = 10 early development the same 6 early development the same 1 embryo with a distinct foot, head and vis- ceral mass own shells 25 early development stones 17 shell-covered embryos own shell 1 early development stones 3 shell-covered embryos stones 10 embryos were already developed and hatched a few days later REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 65 temperature is 1460.55 for 10 m depth, and 1342.65 degree-days for 20 m depth.) Liobaicalia stiedae: embryonic develop- ment in culture for capsules laid in September equalled 1080 degree-days, and the young hatched in March. Summed temperature in the lake at 20 m depth from September to March inclusive was calculated to be 718 de- gree-days. Thus, under natural conditions the young should be expected to hatch as late as July. Parabaikalia florii florii: embryonic develop- ment in culture lasted from November to June, and equalled 1440 degree-days. The temperature sum in the lake at 5 m depth from November to June was calculated to be 480.3 degree-days. Consequently in nature, embry- onic development in capsules laid in early No- vember may last 11 mo, and the young will hatch in mid-October or later. For the same egg-laying time, embryogenesis has been calculated to last 11 mo at 10 m depth, and nearly 12 mo at 20 m depth. While embryonic development in clutches laid at 5 m depth in early June may progress faster than in cap- sules laid in November, that is, 8-9 mo (the young hatching in March-April), it is effec- tively the same for 10 m and 20 m depths: 11.5-12.5 mo (the young hatching in May- June). These calculated values of embryonic de- velopment may not coincide with what really occurs in nature, because (a) near-bottom water temperatures at depths below 20 m (35 m or 50 min some areas) depend strongly on bottom relief and, despite being affected by seasonal changes, these temperatures are al- ways colder than at the surface (Rossolimo, 1957); and (b) water temperature variations may be important for embryonic development in nature, and so the constant water tempera- ture in our experiment may have had a nega- tive effect on the overall duration of embryo- genesis. Egg Capsule Morphology Morphologically, egg capsules of the 15 baicaliid species examined can be subdivided into four groups. (1) Single lens-shaped capsules with con- vex upper surface: On a smooth substratum, the lower surface is flat, but when laid in the umbilicus of another mollusc (Megalovalvata, Valvatidae; Choanomphalus, Planorbidae) or in a cavity in a stone, the lower part of the cap- sule is convex. The margin and the basis is mat and whitish, while the top is transparent. This type of capsule is laid by Liobaicalia stiedae (Fig. 4B), Maackia herderiana, M. variesculpta, M. bythiniopsis (Fig. 5A), M. costata, Teratobaikalia ciliata (Fig. 5B), and Pseudobaikalia zachwatkini (Fig. 4C). Cap- sules of M. bythiniopsis are dark-brown, not transparent and have a pleated surface; cap- sules of the other species are smooth and lighter-coloured (light-yellow or light-brown). Thus, the capsules of different species differ in their overall sizes and the sizes of their mar- gins. (2) Cap-like capsules with no or small mar- gins, often in clusters: Their upper surface is hemispherical, the lower concave or almost flat (depending on the substratum the cap- sules are laid on). These capsules are smooth, and the top is more or less transpar- ent. The capsule color varies from light-yellow to light-brown. The basal part including the margin is mat and whitish. This type of cap- sule is laid by Parabaikalia oviformis (Fig. 4D), Р Пот, P. elata elata, Baicalia carinata and Godlewskia pulchella. (3) Cup-shaped capsules, single or in clus- ters: These are relatively high, with an im- pressed top, the inner surface being dark or light. This type of capsule is attached to rocks and stones and laid by Baicalia dybowskiana (Fig. 5C) and B. turriformis (Fig. 4A). (4) Single “oval bag”, not attached to the substratum. This type of capsule is produced by Teratobaikalia macrostoma, is smooth, greyish green, almost transparent, and is laid on stones between strands of small algae (Fig. 5D). The sizes of the egg capsule of some baicaliid species are given in Table 5. Protoconch Protoconchs have been examined in 26 species, but this paper discusses only the 18 for which adequate numbers could be mea- sured. There are two groups of protoconchs. The first includes species with distinctly sculp- tured shells having 10-25 spiral lirae, for ex- ample: Teratobaikalia ciliata (Fig. 9H-K), T. duthiersii (Fig. 9A-E), T. macrostoma (Fig. 9F, G), T. nana group (Fig. 10A-H), Maackia herderiana (Fig. 7B-D), M. variesculpta (Fig. 7E-J), M. costata (Fig. 8A-C), Pseudobaikalia contabulata, Ps. zachwatkini (Fig. 6G-J), Li- obaicalia stiedae (Fig. 6A-F), and species of Korotnewia. Protoconchs of the second group have a malleate microsculpture and only spi- 66 SITNIKOVA, ROPSTORF 8 RIEDEL FIG. 4. Egg capsules of baicaliid species. A, Baikalia turriformis— both in frontal view. B, Liobaicalia stiedae— frontal and apical view. C, Pseudobaikalia zachwatkin cluster in apical view. (Scale bars equal 1 mm). i- frontal and apical view. D, Parabaikalia oviformis— REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 67 FIG. 5. Egg capsules of baicaliid species, frontal and apical views. A, Maackia bythiniopsis. B, Teratobaikalia ciliata. C, Baicalia dybowskiana. D, Teratobaikalia macrostoma. (Scale bars equal 1 mm). 68 SITNIKOVA, ROPSTORF & RIEDEL TABLE 5. Egg capsule sizes (mm) of some baicaliids. Mean + standard deviation (range). Species (number of specimens) Liobaicalia stiedae (n = 10) Pseudobaikalia zachwatkini (n = 18) Maackia variesculpta (n = 20) M. herderiana (n = 4) M. bythiniopsis (n = 14 for diameter, п = 5 for height) Teratobaikalia macrostoma (n = 3) T. ciliata (n = 6) Parabaikalia florii florii (n = 4) P. florii kobeltiana (n = 4) P. oviformis (n = 11) P. elata elata (n = 8) Baicalia dybowskiana (n = 20) B. turriformis (n = 3) B. carinata (n = 13) Godlewskia pulchella (n = 6) Diameter Height maximal minimal 1.21. = 0.10 1.9 + 0.05 0.52 + 0.07 (1.04-1.25) (1.0-1.15) (0.42-0.60) 1.27 + 0.09 112082 0.63 + 0.08 (1.11-1.40) (1.10-1.35) (0.50-0.85) 1.38 + 0.08 12730107 0.60 +0.07 (1.30-1.50) (1.10-1.35) (0.50-0.80) 1.40 + 0.04 1.00 + 0.09 0.34 + 0.06 (1.35-1.70) (0.80-1.20) (0.30-0.50) 1.75 = 0:16 1861 0510, 1.09 + 0.162 (1.50-2.00) (1.42-1.80) (0.8-1.19) 1.52 + 0.06 113722210103 0.95 + 0.03 (1.55-1.7) (1.3-1.4) (0.95-1.0) 162.2 0412 1.54 + 0.04 0.83 + 0.12 (1.65-1.68) (1.48-1.58) (0.73-1.00) 1:52 0104 1.32 =/0:03 1.27 + 0.04 (1.48-1.55) (1.30-1.37) (1.22-1.30) 1.83. =/0:05 1.60 + 0.00 1.04 + 0.08 (1.8-1.9) (1.60-1.60) (0.95-1.10) 1.41 + 0.06 1:23 10.08 0.94 + 0.08 (1.33-1.48) (1.11-1.37) (0.85-1.11) 1.36 + 0.09 1.24 + 0.09 0.92 + 0.11 (1.23-1.48) (1.11-1.41) (0.74-1.10) 2.07 = 0.12 1.42 + 0.22 1.20 + 0.18 (1.80-2.25) (1.00-1.75) (0.90-1.60) 1.87 + 0.15 1.50 + 0.26 15030540 (1.7-2.0) (1.3-1.8) (1.40-1.60) 1.66 + 0.08 1252230314 0.94 + 0.09 (1.54-1.79) (1.15-1.70) (0.77-1.00) 1.44 + 0.09 1.23 + 0.04 0.80 + 0.08 (1.35-1.60) (1.20-1.30) (0.75-0.90) ral striae but no lirae for example, M. bythin- iopsis (Fig. 8D-F), Godlewskia pulchella, G. columella, B. dybowskiana (Fig. 12A-D), Baicalia carinata (Fig. 12E-G), B. turriformis (Fig. 12H-J), Parabaikalia oviformis (Fig. 11A, C-F), P. elata elata, and P. Пот (Fig. 11G-I), excluding Parabaikalia florii kobeltiana). SEM investigation revealed that species with lirae on the protoconch also may have lirae on the teleoconch (Fig. 7H: M. variesculpta, Fig. 9A: T. duthiersii, Fig: 9H: T. ciliata, Fig. 10E: T. nana group), butthey may become faint on the last whorls. In L. stiedae the lirae continue as rows of periostracal “hairs” (Fig. 11A). A divergence in the angle between proto- conch and teleoconch axes is particularty no- ticeable in Liobaicalia stiedae, where it is sometimes as great as 40° (Fig. 6A-C). Other species examined exhibit only little deviations between the axes, never exceeding 5-10° (Fig. 7H). In some species, it is almost impos- sible to detect an diverging angle. Variation of protoconch size and number of whorls has enabled statistical analysis to be performed for some species (Table 6). Maackia bythiniopsis has the biggest proto- conch diameter (1.305 mm) (figures in paren- theses are mean values) and the Terato- baikalia nana group the smallest (0.625 mm), having also the smallest number of whorls. The shells of the different species of the 7. nana group described by Beckman & Starobogatov (1975) according to one of us (P.R.), may represent only intraspecific varia- tions of T. nana—i.e., forma clandestina (Fig. 10A), forma nana (Fig. 10B), forma producta (Fig. 10C), and forma humerosa (Fig. 10D). The protoconchs of both extremes, forma clandestina (Fig. 10E, F) and forma humerosa (Fig. 10G, H), do not differ neither in dimen- REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 69 FIG. 6. A-F. Liobaicalia stiedae from Kultuk. A, Teleoconch with uncoiled whorls and rows of filaments. B, Nearly planispiral protoconch seen from above. Note the distinct angle to the teleoconch axis. C, Protoconch seen from beneath. D, Hatchling in frontal view. E, Hatchling in apical view. F, Close-up of the nonspiral of the protoconch. G-J, Pseudobaikalia zachwatkini from Kultuk. G, Teleoconch. H, Hatchling in apertural view. |, Hatchling in apical view. (Scale bars: A, G = 2 тт, B-E, Н, | = 200 um, F, J = 50 um). 70 SITNIKOVA, ROPSTORF & RIEDEL FIG. 7. Maackia. A-D, M. herderiana. A, 2-month-old embryo. B, Teleoconch of specimen with well ex- pressed ribs. C, Hatchling in apertural view. D, Hatchling in apical view. E-J, M. variesculpta. E, Typical teleo- conch. Е, Hatchling in apertural view. G, Hatchling in apical view. H, Apex — protoconch tilted to teleoconch axis; spiral lirae of the protoconch continue onto the teleoconch whorls. I, Nonspiral in apical view. J, Magni- fication of spiral lirae. (Scale bars: A, | = 100 um, В = 1 mm, E = 0.5 mm, С, О, F, G = 200 um, J = 10 um). REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 71 FIG. 8. Maackia. A-C, Maackia costata. A, Teleoconch. B, Hatchling in apertural view. C, Hatchling in apical view. D-F, Maackia bythiniopsis. D, Teleoconch. E, Hatchling in apertural view. F, Hatchling in apical view. (Scale bars: A, D = 5 mm, В, С = 200 um, E, F = 500 um). sions nor in sculpture. The highest proto- conch (©: 1.226 mm) with the largest number of whorls (2.6) was found in Baicalia dy- bowskiana (Fig. 12B, C) and the broadest non-spiral part (©: 0.223 mm) in Parabaikalia florii florii. Liobaicalia stiedae has been found to possess the lowest protoconch (0: 0.397 mm, Fig. 6D). Cluster analysis (K-means clustering) has distinguished three groups based on their pro- toconch morphology (Fig. 13). The first group (or cluster) includes Parabaikalia oviformis, P. Пот, P. elata elata, Teratobaikalia macros- toma, Maackia bythiniopsis, two specimens out of six Korotnewia semenkewitschi (see below). The second group includes Baicalia carinata, В. dybowskiana, Godlewskia pul- chella, and Teratobaikalia ciliata. The third group includes Liobaicalia stiedae, Pseudo- baikalia zachwatkini, Ps. pulla, Parabaikalia elata dubiosa, Teratobaikalia duthiersii, the T. nana group, Maackia herderiana, M. varies- culpta, Korotnewia korotnevi), and four speci- mens (out of six) of K. semenkewitschi (see below). Separation into more clusters (4-5) pro- duced a dubious result: specimens of the same species were included in different clus- ters. An effort to divide protoconchs into two clusters resulted in one cluster embracing the whole of the above first and second groups, while the second cluster just overlapped group three. The “joining tree clustering” den- drogram (complete linkage) shows the first two groups close to each other (Fig. 14). The similarity of these groups is manifest when comparing mean values of characters of each species from clusters one and two. For convenience, we will further refer to the three clusters obtained by the name of their 72 SITNIKOVA, ROPSTORF & RIEDEL FIG. 9. Teratobaikalia (Trichiobaikalia). A-E, T. duthiersii. A, Teleoconch —where the spiral lirae cross the axial ribs, they differentiate to periostracal hairs. В, Apex in lateral view — protoconch and teleoconch axes diverge. C, Protoconch in apical view. D, Close-up of spiral lirae. E, Nonspiral appears submerged. F-G, T. macrostoma. F, Conch of a juvenile with aperture characterized by a margin of collar-like plates; umbilicus widely open; transition from protoconch to teleoconch marked by an arrow. G, Nonspiral in lateral view. H-K, T. ciliata. H, Teleoconch of a specimen with well developed ribs and periostracal hairs. |, Hatchling in aper- tural view J, Hatchling in apical view. К. Nonspiral. (Scale bars: А = 500 um, В, C, I, J = 200 um, F, H= 1mm, D = 10 um, E, G= 100 um) REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 73 FIG. 10. Teratobaikalia nana group. A, Teleoconch of Т. nana f. clandestina. В, Teleoconch of Т. nana f. pro- ducta. С, Teleoconch of Т. папа f. nana. D, Teleoconch of Т. папа f. humerosa. E, Е, Protoconch and non- spiral of T. nana f. clandestina. G, H, Protoconch and nonspiral of T. nana f. humerosa. (Scale bars: A, D = 1 тт, В, C=0.5 mm, E, G = 200 um, F, H = 100 um). first species: the first cluster as the Para- baikalia oviformis group, the second as the Baicalia carinata group, and the third as the Liobaicalia stiedae group. All three groups reliably differ by the num- ber of whorls (no overlapping mean values), while the sizes of their non-spiral parts are al- most identical (Table 7). The protoconch type of the Baicalia carinata group species (cluster 2) exhibits the highest number of whorls. Its diameter is close to the Liobaicalia stiedae group (cluster 3), but its height is very close to 74 SITNIKOVA, ROPSTORF & RIEDEL FIG. 11. Parabaikalia. A-F, Parabaikalia oviformis. A, Teleoconch. B, Initial shell formation in a 2-month-old embryo; the posterior visceral mass is covered by a purely organic shell exhibiting a pit-like structure re- flecting the activity of the shell gland. C, Embryonic shell with faint spiral striae and malleate microsculpture. D, Same specimen as in C in apical view. E, Apex of a juvenile which exhibits the transition between embry- onic and postembryonic shell by dense growth increments. F, Nonspiral of the same specimen as in C and D. G, H, Parabaikalia florii. G, Teleoconch. H, Hatchling with faint spiral striae and malleate microsculpture on its shell. |, Hatchling in apical view. (Scale bars: A, G = 5 mm, B, F = 100 um, C, D = 200 um, E, H, | = 500 um) REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 75 FIG. 12. Baicalia. A-D, Baicalia dybowskiana. A, Teleoconch. B, Hatchling in apertural view. C, Hatchling in apical view demonstrating that the first axial ribs are already formed inside the egg capsule. D, Close-up of nonspiral and first whorl showing the shell ornamentation of faint striae and malleate deepenings. E-G, Baicalia carinata. E, Teleoconch. F, Hatchling in apertural view. G, Hatchling in apical view. H-J, Baicalia tur- riformis. H, Teleoconch. |, Hatchling in apertural view. J, Hatchling in apical view. (Scale bars: A, E, H, = 5 mm, others = 100 um). TABLE 6. Protoconch size of some baicaliids (diameter, height, width of nonspiral in mm, N-whorls: num- ber), mean + 6 (range), (number specimens). Species N-whorls Liobaicalia stiedae Teratobaikalia nana f. clandestina T. nana f. humerosa Maackia herderiana M. variesculpta M. bythiniopsis Pseudobaikalia pulla pulla Ps. zachwatkini Ps. contabulata Teratobaikalia duthiersii T. ciliata T. macrostoma Korotnewia semenkewitschi K. korotnevi Parabaikalia elata elata P. elata dubiosa P. oviformis Р florii от Baicalia dybowskiana B. carinata B. carinato-costata Godlewskia pulchella 1.79 + 0.06 (1.75-1.85) (n = 7) 1.50 + 0.055 (1.45-1.6) (n = 5) 1.45-1.5 (n =2) 1.51 10:07 (1.4-1.65) (n = 10) 1.60 + 0.077 (1.45-1.75) (n = 14) 1.87 + 0.063 (1.8-2.0) (m= 10) 1.56==0.225 (1.55-1.6) (n=4) 1.75 + 0.00 (n = 3) 1.58 + 0.063 (1.55-1.75) (n = 10) 2.21 + 0.055 (2.15-2.25) (n=5) 2.08 + 0.084 (2.0222) (n=5) 1.68 + 0.052 (1.65-1.80) (n=6) (n = 10) 1.98 + 0.057 (1.85-2.25) (n = 7) 2.64 + 0.114 (2.55-2.75) (n = 5) 2.40 + 0.092 (2.25-2.60) (n = 23) 2.03 + 0.035 (2.00-2.05) (n = 2) 2.38 + 0.161 (2.2-2.55) (n = 5) Diameter 0.904 + 0.114 (0.78-0.89) (n = 7) 0.625 + 0.018 (0.6-0.64) (n = 5) 0.59-0.64 (n=2) 0.757. =#:0.023 (0.73-0.81) (n = 10) 0.73 + 0.021 (0.67-0.75) (n = 14) 1.305 + 0.061 (1.27-1.42) (n = 10) 0.678 + 0.017 (0.66-0.70) (n = 4) 0.848 + 0.029 (0.80-0.88) (n = 4) 0.685 + 0.042 (0.61-0.75) (n = 10) 0.677 + 0.011 (0.67-0.69) (n = 3) 0.779 + 0.036 (0.73-0.80) (n = 5) Teli = 0.127 (1.04-1.34) (n = 5) 0.850 + 0.061 (0.77-0.97) (n = 6) 0.87 + 0.072 (0.83-0.91) (n = 6) 0.961 + 0.0567 (0.86-1.05) (n = 10) 0.82 + 0.473 (0.80-0.87) (n=3) 1.055 + 0.054 (0.98-1.13) (n = 15) 1. 137 = 0.056 (1.08—1.16) (п= 7) 0.894 + 0.053 (0.84-0.95) (n = 5) 0.837 + 0.056 (0.75-0.99) (n = 23) 0.76 + 0.028 (0.74-0.78) (n=2) 0.638 + 0.042 (0.61-0.71) (n = 5) Height 0.397 + 0.032 (0.37-0.42) (n = 4) 0.423 + 0.028 (0.38-0.45) (n = 5) 0.4-0.43 (n = 2) 0.574 + 0.012 (0.56-0.59) (n= 10) 0.537 + 0.038 (0.45-0.6) (n = 14) 1.004 + 0.069 (0.93-1.14) (n = 9) 0.56 + 0.028 (0.54-0.58) (n = 2) 0.725 + 0.021 (0.71-0.73) (n = 2) 0.555 + 0.026 (0.51-0.59) (n = 10) 0.553 + 0.081 (0.49-0.61) (n = 3) 0.913 + 0.031 (0.87-0.94) (n = 4) 1.226 + 0.085 (1.1-1.31) (n = 5) 0.728 + 0.098 (0.63-0.88) (n = 5) 0.72 + 0.000 (n = 3) 1.018 + 0.0537 (0.93-1.05) (n = 10) 1.020 + 0.073 (0.87-1.14) (п= 15) 1.174 + 0.071 (1111836) (137) 1.226 + 0.029 (1:21=1.27) (n=5) 1.115 + 0.082 (1.06-1.34) (n = 23) 0.855 + 0.007 (0.85-0.86) (п = 2) 0.834 + 0.085 (0.75-0.96) (n = 5) Width of nonspiral 0.104 + 0.040 (0.6-0.165) (n = 6) 0.132 + 0.032 (0.088-0.17) (n = 5) 0.130-0.138 (n = 2) 0.180 + 0.037 (0.9-0.21) (n = 10) 0.147 + 0.020 (0.115-0.19) (n = 14) 0.175 + 0.043 (0.12-0.23) (n = 10) 0.158 + 0.033 (0.110-0.180) (n = 4) 0.182 =0:017 (0.165-0.200) (п= 3) 0.163 + 0.031 (0.104-0.206) (n = 10) 0.127 + 0.032 (0.096-0.160) (n = 3) 0.136 + 0.023 (0.100-0.160) (n = 5) 0.211 + 0.022 (0.180-0.230) (n = 5) 0.196 + 0.027 (0.176-0.245) (n = 6) 0.14 + 0.028 (0.120-0.160) (n = 2) 0.205 + 0.019 (0.180-0.240) (n = 10) 0.205 + 0.009 (0.195-0.213) (n = 3) 0.206 + 0.023 (0.153-0.239) (n = 15) 0.223 + 0.029 (0.185-0.270) (n =7) 0.186 + 0.029 (0.150-0.230) (n = 5) 0.193 + 0.035 (0.107-0.246) (n = 23) 0.185 + 0.021 (0.170-0.200) (n=2) 0.150 + 0.026 (0.123-0.180) (n=5) REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 77 mm (n-whorls - number) —4— Cluster 1 | —O— Cluster 2 —X — Cluster 3 1 O 4 n-w horls diameter Characters height non.spir.part FIG. 13. Plot of means for each cluster (protoconch size in mm). the Parabaikalia oviformis group (cluster 1). The L. stiedae and P. oviformis groups differ in the mean values of all characters. DISCUSSION Reproduction Prior to this study, data on baicaliid repro- duction could be found in one article only (Gavrilov, 1953). Based on egg capsules at- tached to stones lifted monthly from a depth of 3 m near Listvenichnoe (18 km south of Bol- shie Koty). Gavrilov concluded that Maackia herderiana reproduced all the year round and had no seasonality, but Table 8 shows two re- production peaks for this species: Septem- ber-October and March-May. Our data confirm that in Baicaliidae in gen- eral the reproduction continues and egg cap- sules were produced throughout the year, even in January. It is suggested that M. herde- riana does have two reproduction peaks: (1) spring-first half of summer, and (2) second half of summer-early autumn. (Maackia vari- esculpta may also have two peaks in repro- duction.) Capsules found in summer and autumn with embryos at different development stages, and calculated duration of embryoge- nesis do not refute our hypothesis of two re- production maxima in littoral species. How- ever, it is not known how long the egg-laying period lasts in one female and how fecund it is, nor do we know if individual females lay eggs twice or only once a year. In the latter case, two generations —the spring-summer and the autumn—may be hypothesised to exist within these populations, with different egg-laying times. Besides, the reproductive period in one individual may be long enough for egg capsules at the end of one period to coincide in time with the capsules laid at the beginning of the other period. Our investigations show that the period when the young hatch in great numbers is re- stricted to two different seasons: spring-early summer and autumn. How can this be ex- plained? Earlier experiments by Sitnikova (unpubl. data) demonstrated that adult bia- caliids placed in aquaria with Lake Baikal water and substrata (stones with vegetation, sand with detritus) in September-October re- mained active at temperatures of 4-6°C with no food added until April; then all molluscs died, including the young that had been mov- ing along aquarium walls 10-15 days after hatching. Besides, the shells of littoral mol- luscs from the lake start to grow in spring, 78 0.4 SITNIKOVA, RÓPSTORF & RIEDEL Euclidean distance 0.6 0.8 I 1.2 1.4 1.6 FIG. 14. Dendrogram of 144 specimens using protoconch size TABLE 7. Protoconch in the tree groups in the cluster analysis (size in mm), mean + standard deviation (range). Protoconch characters N-whorls Diameter Height Width of nonspiral reaching 0.5-2.0 mm in diameter in May-July. These observations suggest that the gas- tropods may need additional food in spring, and this food becomes available in the spring- summer period. According to Gavrilov (1953), M. herderiana 0.6 0.8 | Linkage distance 1.89 + 0.15 (1.6-2.25) 1.12+0.14 (0.86-1.42) 1.08 + 0.12 (0.87-1.36) 0.2 + 0.04 (0.12-0.27) Cluster 1 (n = 49) 1.2 Cluster 2 (n = 37) 1.4 is a bacterio-feeder, and he reported plank- tonic forms to be dominant among diatoms found in the stomachs of M. herderiana. Ex- amination of baicaliid distribution on stones and rocks in the Zhilische canyon (Bolshie Koty Bay; Fig. 1) in relation to rock slope has 2.41 + 0.14 (2.15-2.75) 0.81 + 0.09 (0.61-0.99) 1.07 + 0.14 (0.75-1.34) 0.18 + 0.04 (0.14.11) 1.6 Cluster 3 (n = 58) 1.59 + 0.11 (1.4-1.85) 0.73 + 0.08 (0.59-0.91) 0.55 + 0.09 (0.37-0.74) 0.16 + 0.08 (0.09-0.96) REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 79 TABLE 8. Number of Maackia herderiana egg cap- sules laid in different periods (after Gavrilov, 1953). Date stones Date stones Number of placed on the raised up from egg capsules bottom the bottom on stones 1 July 1947 2 August 1947 1 5 August, 1947 31 August 1947 7 3 September 1 October 1947 59 1947 6 October 1947 3 November 1 1947 13 November 6 December 3 1947 1947 17 December 7 January 1948 9 1947 15 January 1948 5 February 1948 12 12 February 23-24 March 18 1948 1948 21-25 March 21-24 May 38 1948 3 June 1948 2-3 July 15 revealed a positive correlation between the abundance and biomass of this species and slope. This observation suggested M. herderi- ana may be a suspension-feeder (Kamaltynov etal., 1998), that is, it could assimilate not only bacteria but all the nutrients in the near-bottom water layer (i.e., detritus and phytoplanktonic sediments). We did not specifically study baicaliid feeding, but random dissection of stomachs revealed planktonic and benthic di- atoms, detritus, sponge spicules, and white amorphous substance resembling yeast. Sometimes one component prevailed. Long-term observations (Kozhov, 1963; An- tipova, 1974) revealed two periods in phyto- plankton development in Lake Baikal, when its abundance and hence, biomass, in- creased: spring (April-May, maximum), and autumn (September-October, sometimes later). These periods may shift slightly from year to year, but their seasonality is constant. Bacterioplankton has also been found to ex- hibit two maxima in their development (Kozhov, 1963). Planktonic diatoms and bacteria sinking to the bottom of Lake Baikal may provide the necessary food for suspension-feeding mol- luscs. Plant detritus and microphytobenthos in the diet of molluscs that collect benthic food should not be ignored either. According to Vot- intsev (1961), the influx of organic matter into the Lake Baikal increases considerably during the spring and summer floods. Microphyto- benthos has been found to exhibit two peaks in the development of its biomass and abun- dance: the smaller occurs during the under- ice period (February), the larger takes place in summer (July) (Pomazkina, 2000). Increased content of both, detritus in bottom sediments and microphytobenthos, may act as a stimu- lus for the hatching of species feeding on de- tritus and microphytobenthos. These species include, for example, Parabaikalia elata, P. oviformis and P. florii, which collect their food directly from the lake bottom. Gastropods living in “soft” waters with low calcium content (up to 15mg/l in Lake Baikal, according to Votintsev, 1961), are known to consume calcium from water plants in order to build their shells (Young, 1975). This fact may indicate that Lake Baikal molluscs satisfy their calcium requirements by consuming phyto- plankton and phytobenthos. The available ev- idence suggests that hatching of the young may be timed specifically to coincide with in- creased vegetable resources. Egg Capsules and Protoconch Correlation analysis of all species exam- ined showed (Table 9) that the minimal diam- eter of egg capsules (margins excluded) cor- relates with the protoconch diameter (r = 0.73) and the capsule height generally correlates with the protoconch height (r = 0.82). The larger the protoconch, the greater volume it occupies in the capsule. But in several species — Pseudobaikalia zachwatkini, Tera- tobaikalia ciliata, Godlewskia pulchella, Parabaikalia oviformis, P. elata elata — the protoconch is higher than the egg capsule. Observation of the young of these species re- vealed that the molluscs either lie or lean in- side their capsules and thus make maximum use of the capsule’s volume. How the protoconch size influences teleo- conch size is uncertain. The answer requires investigation of the teleoconch of each spe- cies, and sampling must be statistically reli- able. Based on shell size data reported by Kozhov (1936), we may ascertain the follow- ing: (1) In the species investigated, the teleo- conch is, on average, 13 times higher (minimum in Teratobaikalia nana f. clan- destina and T. macrostoma, maximum in Liobaicalia stiedae), six times wider (mini- mum in Godlewskia pulchella, maximum in Parabaikalia Пот! Пот) and has three times more whorls (minimum in Terato- baikalia ciliata, maximum in Maackia herderiana) (Table 10) than the proto- conch. This means that height and width 80 SITNIKOVA, ROPSTORF & RIEDEL TABLE 9. Correlation of protoconch and egg capsule size. Marked correlations are significant at p < 0.05, mean + standard deviation (range), in mm. m + Ô m + Ô (range) (range) Characters (n=75) r(X, Y) tvalue Characters (n = 95) r(X, Y) t-value egg capsule height 0.88 + 0.24 eggcapsule 1.17 + 0.23 (0.30-1.60) diameter (1.04-2.25) protoconch height 0.90 + 0.25 0.82 12.08 protoconch 0.91 = 0.20 0.73 10.2 (0.37-1.136) diameter (0.59-1.42) TABLE 10. Comparison of teleoconch and proto- conch average size (H, п: height, W, w: width, N, п: number of whorls, capital letters refer to the teleo- conch, lower case to the protoconch); size of teleo- conch after Kozhov (1936). Species H/h W/w N/n Liobaicalia stiedae 25.3 Hal РИ Pseudobaikalia zachwatkini 10.4 4.5 32 Ps. contabulata 10.8 4.4 3.5 Ps. рийа tenuicosta 9.9 4.2 3.7 Maackia costata 14.6 7.65 3.4 M. herderiana ESA 6.8 3.8 M. variesculpta 12.5 5.5 3.5 M. bythiniopsis 10.1 5.6 2.8 Teratobaikalia macrostoma 7.8 6.7 2.2 T. nana f. clandestina 7.8 See 27, T. duthiersii 1077 5.8 Sul T. ciliata 9.4 5.6 2.4 Parabaikalia elata dubiosa 9.8 4.6 ZT: P elata elata les 6.2 3:0) P. oviformis 13.9 7.9 383 P. florii florii 19.7 11.1 Sal Korotnewia semenkewitschi 11.9 5.4 3.6 K. korotnevia 24.8 8.4 4.7 Baicalia carinata 17.8 7.9 3.6 B. dybowskiana 8.7 4.6 2.6 Godlewskia pulchella 10.4 4.4 32 do not increase uniformly with an increas- ing number of whorls. Species assigned to one genus and simi- lar in teleoconch size, are also similar in protoconch size, and the growth of their teleoconchs differ very slightly, for exam- ple, in Maackia herderiana and M. varies- culpta, M. costata, Parabaikalia elata elata and P. oviformis. (3) Species with similar protoconch sizes may differ considerably in teleoconch height and width, for example, Parabaikalia florii florii and P. oviformis. The former is larger and its teleoconch is 20 times higher than the protoconch, compared to only 14 times in the latter species. (4) Parabaikalia elata elata and Р elata du- biosa differ in teleoconch and protoconch D sizes (cluster analysis places them into different groups), and their definitive shell growth proceeds differently. We have distinguished four groups of baicaliid egg capsules; and three size-groups of protoconchs. Capsule and protoconch sizes correlate. Seven species — Liobaicalia stiedae, Maackia variesculpta, M. herderiana, M. bythiniopsis, M. costata, Pseudobaikalia zachwatkini, and Teratobaikalia ciliata—have lens-shaped capsules (Table 11). Five ofthem have been pooled together by embryonic shell size (cluster analysis) into cluster three (Liobaicalia stiedae group). Two species are exceptions: Teratobaikalia ciliata, assigned to cluster two (Baicalia carinata group) due to its protoconch size and large egg capsules, and M. bythiniopsis, also having large capsules, but these are dark-brown in color, with a pleated surface, and are assigned to cluster one (Parabaikalia oviformis group) based on protoconch size. The same cluster includes Teratobaikalia macrostoma based on proto- conch ratios, though this species lays oval, bag-shaped capsules. Species grouped to- gether by protoconch size into cluster one (Parabaikalia oviformis, P. elata elata, P. florii flori) have cap-shaped capsules. Finally, species pooled together by protoconch size into cluster two (Godlewskia pulchella, Baicalia dybowskiana, B. carinata and Terato- baikalia ciliata), have, with the exception of the last two, cup-shaped capsules. Taxonomic Remarks Size differences of egg capsules of two subspecies, Parabaikalia florii florii and P. florii kobeltiana, and of protoconchs of P. elata elata and P. elata dubiosa suggest the possi- ble existence of independent allopatrical species, but this must be tested by further analysis. Parabaikalia florii Пот! and P. elata elata inhibit southern and middle parts of Lake REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 81 TABLE 11. Comparison of protoconch size and type of egg capsules of some baicaliids. Species Parabaikalia oviformis P. florii По! P elata elata Baicalia carinata Godlewskia pulchella Baicalia turriformis B. dybowskiana Teratobaikalia macrostoma T. ciliata T. duthiersii Maackia herderiana M. variesculpta M. costata M. bythiniopsis Protoconch-size cluster 1 (middle size) 1 (middle size) 1 (middle size) 2 (large) 2 (large) not determined 2 (large) 1 (middle size) 2 ( 3 (small) not determined 1 (middle size) Type of egg capsulae cap-like the same the same the same the same cup-like the same oval bag lens-shaped, lighter colours, smooth unknown lens-shaped, lighter colours, smooth the same the same lens-shaped, dark-brown, Pseudobaikalia zachwatkini Liobaicalia stiedae 3 (small) Baikal, whereas Р florii kobeltiana and P. elata dubiosa were found in northern Lake Baikal. We suggest that the reason the proto- conchs of Korotnewia semenkewitschi from different regions of Lake Baikal fall into differ- ent clusters may be explained by this actually being two species. Judging from teleoconch variability it is possible but requires proving. Morphological similarity of protoconchs in the species examined of the genera Baicalia and Godlewskia — cluster analysis assigns them into the same cluster, height/width ratio (Table 12) does not differentiate between them reliably — allows to include them in one genus, but, perhaps, as independent subgen- era, in view of the differences in the structure of their female reproductive systems. The species of Baicalia have a wide renal oviduct loop located on the ventral and ventrolateral surface of the gonoduct, whereas species of Godlewskia have a narrow loop that runs downwards to the dorsal side (Sitnikova, 1991b). Of the three species (Maackia bythiniopsis, M. herderiana and M. variesculpta) assigned by Lindholm (1909), Kozhov (1936) and Sit- nikova (1991b) to the subgenus Eubaicalia Lindholm, 1924, the first species has a larger protoconch lacking the lirae found in the other two species. Cluster analysis of variable char- acters places the protoconch of M. bythiniop- 3 (middle size) pleated surface lens-shaped, lighter colours, smooth the same sis into the Parabaikalia oviformis group, and its size and sculpture are also similar to P. ov- iformis (and to the genus Parabaikalia, in general), apparently suggesting a relationship between these species. However, the teleo- conch of M. bythiniopsis, as in other species of the subgenus Eubaicalia, shows an aper- ture that is distinctly angulated anteriorly (e.g., Fig. 7), which represents an assumed apo- morphy of the subgenus. One of us (T. S.) at- tributes the presence of such an angle to an adaptation to laying egg-capsules in cavities of stones or rocks. The three species exam- ined of the subgenus Eurbaicalia dwell on hard substrata. Maackia herderiana and M. variesculpta lay small capsules is small cavi- ties, whereas M. bythiniopsis lays larger egg capsules in correspondingly larger cavities. The structure of the female reproductive sys- tem of M. bythiniopsis is similar to M. herderi- ana and M. variesculpta, the renal oviduct loop covers half of the surface of the albumen gland, whereas in the species of the genus Parabaikalia, this loop is smaller and covers a third of the surface of the albumen gland (Sit- nikova, 1991b). T-test pairwise comparison of the protoconch proportions of M. bythiniopsis, M. herderiana, M. variesculpta and Para- baikalia oviformis (Tables 11, 12) show M. by- thiniopsis to differ (p < 0.05) from other species in whorl number/height and whorl number/width ratios, but height/width ratio 82 SITNIKOVA, ROPSTORF 8 RIEDEL TABLE 12. Protoconch ratios (п - height, м - width, п - whorl number). h/w mean + à n/h mean + à n/w mean + à Species (range) (range) (range) Liobaicalia stiedae 0.47 + 0.61 4.62 + 0.46 2.18 = 0.13 (0.41-0.53) (4.07-5.00) (2.08-2.37) Teratobaikalia nana f. clandestina 0.69 + 0.04 3.51 = 0.19 2.41 + 0.10 (0.63-0.73) (3.33-3.81) (2.34-2.58) T. nana f. humerosa 0.68 + 0.07 3.56 = 0.27 2.40 + 0.08 (0.63-0.73) (3.37-3.75) (2.34-2.46) Maackia herderiana 0.76 + 0.01 2.62 + 0.04 1.99 + 0.06 (0.72-0.78) (2.50-2.79) (1.89-2.07) M. variesculpta 0.74 + 0.06 2.98 + 0.25 2.20 = 0312 (0.61-0.85) (2.74-3.70) (2.00-2.40) M. bythiniopsis 0.76 + 0.59 1.87 = 0.13 1.42 = 0.10 (0.68-0.86) (1.71-2.11) (1.34-1.46) Pseudobaikalia pulla pulla 0.84 + 0.036 2.77 + 0.14 2.33 = 0:03 (0.80-0.88) (2.67-2.87) (2.31-2.35) Ps. zachwatkini 0.83 + 0.01 241 = 0:07 2.01.=.0:03 (0.83-0.84) (2.36-2.46) (1.99-2.03) Ps. contabulata 0.81 + 0.04 2.85 + 0.18 2.32 + 0.18 (0.73-0.85) (2.62-3.18) (2.09-2.54) Teratobaikalia duthiersii 0.82 + 0.11 2.93 + 0.42 2.37. 20415 (0.69-0.88) (2.54-3.37) (2.25-2.54) T. ciliata 1.18 + 0.01 2.44 + 0.12 2.89 + 0.18 (1.16-1.25) (2.31-2.59) (2.69-3.09) T. macrostoma 1.05 + 0.08 1.70 + 0.18 1.79.0415 (0.91-1.14) (1.60-1.82) (1.57-1.92) Korotnewia semenkewitschi 0.87 + 0.06 2:27.=:0:30 1.96 + 0.15 (0.79-0.93) (2.0-2.62) (1.80-2.14) K. korotnevi 0.82 + 0.04 2290.12 1.88 +0.11 (0.79-0.87) (2.2-2.4) (1.76-1.92) Parabaikalia elata elata 1.06 + 0.065 1.76 + 0.10 1:86 10115 (0.95-1.14) (1.62-1.93) (1.63-2.10) P. elata dubiosa 0.84 + 0.042 2.37. = 0318 1.99 + 0.01 (0.81-0.87) (2.28-2.46) (1.99-2.00) P. oviformis 0.97 + 0.05 1.74 + 0.11 1:68 8011 (0.84-1.03) (1.54-1.92) (1.49-1.86) Р florii Пот 1.04 + 0.08 1.70 + 0.08 177 == 10:15 (0.89-1.65) (1.56-1.80) (1.6-1.96) Baicalia dybowskiana 1.37 + 0.076 215 2021 2.96 + 0.28 (1.29-1.43) (2.01-2.29) (2.71-3.27) B. carinata 1280106 2416 = 0.15 2.88 + 0.19 (1.23-1.44) (1.90-2.63) (2.52-3.20) Godlewskia pulchella 1.31 + 0.085 2.86 = 057 3.73 210110 (1.19-1.35) (2.71-3.03) (3.49-4.05) does not differentiate this species (p > 0.05) from other two species of the subgenus Eu- baicalia, but from Parabaikalia oviformis (p < 0.05) (Table 13). Three species assigned to the genus Tera- tobaikalia — T. ciliata, T. duthiersii and T. macrostoma — also require discussion. Ear- lier, these species were assigned to inde- pendent subgenera of the genus Baicalia: T. macrostoma to subgenus Teratobaikalia, T. ciliata to subgenus Dybowskiola, and T. duthiersii to the subgenus Trichiobaikalia (Lindholm, 1909; Kozhov, 1936). Later, these three species were assigned to the genus Ter- atobaikalia based on the morphology of the fe- male reproductive system, but were divided into two subgenera: Teratobaikalia with T. macrostoma, and Trichiobaikalia with 7. ciliata and 7. duthiersii (Sitnikova, 1991b). The pro- toconchs of these species differ in size, and analysis of their protoconch characters places them in different clusters. However, similar sculpture and protoconch shape suggest an affinity of these species, with T. ciliata and 7. macrostoma being the most similar. This is also confirmed by the height/width ratio of the REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 83 TABLE 13. Comparison of protoconch ratios in Maackia bythiniopsis with two other species of Maackia and Parabaikalia oviformis using T-test (h - height, w - width, n - whorl number). Maackia bythiniopsis (n = 10) h/w n/h n/w Maackia herderiana (n = 10) 10:3, Nis. t 14.4, p 0.00 t 24.10, p 0.00 M. variesculpta (n = 14) 223, mis: t 12.01, p 0.00 t 188.99, p 0.00 Parabaikalia oviformis (n = 15) t 9.41, p 0.00 t 2.50, p 0.02 t 6.93, p 0.00 protoconch, which does not differentiate the species (p > 0.05). The fact that T. duthiersii differs (p < 0.05) from T. macrostoma in three of the protoconch ratios used (height/width, whorl number/height and whorl number/ width), and that 7. ciliata and T. duthiersii do not differ in whorl number/height ratio (p > 0.05), indicates that 7. duthiersii is closer to T. ciliata in protoconch morphology than to T. macrostoma. The three species also have one feature in common: the protoconch sculpture that continues as lirae on the teleoconch, dif- ferentiates to periostracal hairs in T. ciliata, plates in T. macrostoma, and “brushes” in 7. duthiersii (Fig. 9A, F, H). CONCLUSIONS Our results allow us to suggest that (1) baicaliids reproduce throughout the year, with two main egg-laying periods (late spring and late summer-autumn); (2) the duration of em- bryogenies depends on egg-laying time for 5 m, 10 m and 20 m depths, where seasonal temperature varies; (3) that the young hatch in greater numbers twice a year, which for suspension-feeder species appears to be timed in coincidence with peaks of phyto- and bacterioplankton production, and for species that consume microphytobenthos and detri- tus, to production peaks in the former, and in- creased input of the latter. While there is a certain degree of specula- tion in our reasoning, we hope that our report will encourage research in order to examine the link between reproduction, hatching and feeding of Lake Baikal endemic molluscs. Morphologically, the egg capsules of the 15 baicaliid species examined can be subdivided into four groups: (1) single lens-shaped cap- sules with a convex upper surface, (2) cap- like capsules with no or narrow margins, often in clusters, (3) cup-shaped capsules, in clus- ters or single, and (4) single “oval bags”, not attached to the substratum. Protoconchs have been examined in 26 baicaliid species. The ontogenetic continua- tion of the lirae on the teleoconch can be de- tected only in a few species. The protoconchs of Baicaliidae (including Liobaicalia stiedae) are orthostrophic, but in L. stiedae the proto- conch axis deviates significantly from the teleoconch axis. Three groups of baicaliids were distinguished on the basis of a cluster analysis of protoconch morphology, and the results do not fully coincide with currently ac- cepted taxonomy of the Baicaliidae. Some species currently assigned to the same genus (or subgenus) differ either in protoconch size or egg capsule morphology. Additional studies of morphology, taxon- omy, population variability of the shell and ecology of the Baicaliidae will provide further insights into the origin and evolution of this group in Lake Baikal. ACKNOWLEDGEMENTS The authors are sincerely grateful to the di- rector of the Limnological Institute of the Siberian Division of the Russian Academy of Sciences (RAS) Prof. M. A. Grachev for mak- ing it possible to conduct the work under his auspices, to the former and the current direc- tor of the Institute of Toxicology (City of Baikalsk), Dr. A. M. Beym (deceased) and Dr. E. |. Grosheva respectively, to the scientist of the latter institute, S. Kudelin, for having suc- cessfully managed experimental work in the aquarium; to the divers V. Votyakov and S. Selyandin, who self-denyingly lifted molluscs from the bottom of Lake Baikal heedless of the weather, and to the captains of the re- search vessels Titov, Vereshchagin, and Obruchev. We are sincerely grateful to M. Bu- lang-Lorcher (Freie Universitat Berlin) for drawing most of the egg-capsules, to Dr. M. N. Shimaraev for consultation with regard to Baikalian water temperature, Drs. G. F. Mazepova, Z. V. Slugina, R. M. Kamaltynov, 84 SITNIKOVA, RÓPSTORF & RIEDEL D. Y. Sherbakov (all Limnological Institute of RAS in Irkutsk), Dr. Y. I. Starobogatov and Dr. V. G. Sideleva (Zoological Institute of RAS in St. Petersburg) for their kind reading of the manuscript and comments. Many thanks for a financial support from the Jessup-Foundation of the Academy of Natural Science of Philadelphia (ANSP) and for the kindness of Dr. C. Goulden (Director of the Institute for Mongolian Biodiversity and Ecological Stud- ies, ANSP). The first author was able to study contemporary literature in the malacological library of the ANSP and to complete the man- uscript. We are particularly indebted to Dr. G. M. Davis (ANSP) for his critical remarks at the various stages of the progress of the manu- script. We also thank two anonymous review- ers for their comments on the manuscript. The work was supported by the Deutsche Forschungsgemeinschaft (DFG) project Ri 809/1 (Dr. Riedel), the Russian Foundation of Basic Research (RFBR) grant N 97-04-96237 (Dr. Timoshkin), RFBR grant N97-04-63036k (Prof. Grachev), RFBR grant N98-04-49276a, grant N98-04-63058k (Dr. Sitnikova) and by a Japanese grant (Prof. Wada): “An Integrative Study on Biodiversity Conservation under Global Change and Bioinventory Manage- ment System”. LITERATURE CITED ANTIPOVA, М. L., 1974, Annual fluctuation of phy- toplankton in the area of Bolshiye Koty in the pe- riod of 1960-1970. Pp. 75-84, in: O. M. KOZHOVA, ed., Productivnost' Baikala i antropogennye izmemeniya ego prirody [Productivity of Lake Baikal and anthropogenic changes of its nature], Ministerstvo vysshego i srednego obrazovaniya RSFSR, Irkutskiy gosudarstvennyi universitet, Irkutsk (in Russian). BECKMAN, M. Y., & Y. |. STAROBOGATOV, 1975, Baikal deepwater molluscs and their related forms. Pp. 92-111, in: News on the fauna of Baikal, part 1. Trudy Limnologicheskogo Instituta SO AN SSSR, 18(38) (in Russian). DYBOWSKI W., 1875, Die Gastropoden-Fauna des Baikal-Sees, anatomisch und systematisch Bear- beitet. Memories de l’Academie Imperiale des Sciences de St. Petersbourg, 22(8): 1-73, 8 pls. GAVRILOV, G. B., 1953, The reproduction of the Baikalian mollusc Baicalia herderiana and others. Zoologicheskiy Zhurnal, 32(5): 840-843 (in Russian). KAMALTYNOV, R. M., E. B. KARABANOV, T. Y. SITNIKOVA & V. N. ATAMANCHUKOV, 1998, Biota of the underwater rock slopes —new com- ponent of the Baikalian ecosystem. Ekologich- eskie problemy basseinov krupnyh rek (Ecologi- cal problems in basins of large rivers), 203, ab- stract (in Russian). KOZHOV, M. M., 1936, Mollusques du lac Baikal. Systematique, distribution, ecologie, quelques donnies sur la genese et histoire. Akademia Nauk SSSR, Trudy Baikal’skoi Limnologicheskoi Stantsii [Travaux de la Station Limnologique du Lac Bajkal], 8: 1-350, 13 pls. (in Russian, Ger- man summary). KOZHOV, M. M., 1963, Lake Baikal and its life. Monographiae Biologicae, 11: 1-344. LINDHOLM, W. А., 1909, Die Mollusken der Baikal- Sees. Wissenschaftliche Ergebnisse einer Zoolo- gischen Expedition nach dem Baikal-See unter Leitung des Professors Alexis Korotneff in den Jahren 1900-1902, 4: 1-104, 2 pls. LINDHOLM, W. A., 1924, Collectanea baicalica |. Archiv fur Molluskenkunde, 56(6): 217-225. POMAZKINA, С. |., 2000, Microphytobenthos of the southern Baikal. Abstract of the PhD thesis, Insti- tute of Geography, SD of RAS, Irkutsk, 1-20 (in Russian). | PONDER, W. Е. & A. WAREN, 1988. Appendix: Classification of the Caenogastropoda and Het- erostropha —a list of the family-group names and higher taxa. Malacological Review, Supplement 4: 288-326. RIEDEL, F., 1993, Early ontogenetic shell formation in some freshwater gastropods and taxonomic implications of the protoconch. Limnologica, 23(4): 349-368. ROSSOLIMO, L. L., 1957, The thermal regime of Lake Baikal. Trudy Baikalskoy Limnologicheskoy Stantsii AN SSSR, 16: 1-551 (in Russian). SITNIKOVA, T. Y., 1991a, Results of study of egg- clusters of endemic Baikalian mollusks. Pp. 61-73, in: Y. I. STAROBOGATOV, ed., Pazmnozhe- nie i kladki yaits mollyuskov [Reproduction and egg-clusters of molluscs], Trudy Zoologich- eskogo Instituta AN SSSR, Vol. 228, Leningrad (in Russian). SITNIKOVA, T. Y., 1991b, New structure of Baikal endemic molluscs of the family Baicaliidae. Pp. 266-281, in: A. A. LINEVICH & E. L. AFANASIEVA, eds., Morfologiya | evolyutsiya bespozvonoch- nykh [Morphology and evolution of invertebrates], Nauka, Novosibirsk (in Russian). SITNIKOVA, T. Y., 1994, Recent views on the his- tory and diversity of the Baikalian malacofauna. Archiv fur Hydrobiologie, Ergebnisse der Lim- nologie, 44: 319-326. STAROBOGATOV, Y. |. & Т. Y. SITNIKOVA, 1983, The system of the Order Littoriniformes (Gas- tropoda Pectinibranchia). Pp. 18-22, in: 1. M. LIKHAREV, ed., Mollusci, sistematika, ekologiya ! zakonomernosti raspredeleniya. [Molluscs, their systematics, ecology and distribution], Nauka., Leningrad (in Russian). VERBOLOV, V. 1., У. M. SOKOLNIKOV 8 M. М. SHIMARAEV, 1965, Hydrometeorological regime and temperature balance of Lake Baikal. Nauka, Moscow-Leningrad, 373 pp. (in Russian). VOTINTSEV, K. K., 1961, Hydrochemistry of Lake Baikal. Trudy Baikal’skoi Limnologicheskoi REPRODUCTION AND EMBRYOGENY IN BAICALIID RISSOOIDEANS 85 Stantsii [Proceedings of the Baikalian Limnologi- two freshwater species of Gastropoda. Proceed- cal Station], 20: 1-311 (in Russian). ings of the Malacological Society of London, 41: VOZNESENSKY, A. V., 1908, Outline of the climatic 439-445. peculiarities of Lake Baikal. Pp. 173-329, in: O. K. DRIZHENKO, ed., Lotsia / fiziko-geograficheskiy Revised ms. accepted 29 July 2000 ocherk ozera Baikal [Maps and physical and ge- ographical outline of Lake Baikal], St. Petersburg (in Russian). YOUNG, J., 1975, A laboratory study using *Ca tracer, on the source of calcium during growth in MALACOLOGIA, 2001, 43(1-2): 87-101 A GENETIC APPROACH TO RESOLVING TAXONOMIC AMBIGUITY AMONG PLEUROBEMA (BIVALVIA: UNIONIDAE) OF THE EASTERN GULF COAST Karen |. Kandl', Hsiu-Ping Liu?, Robert $. Butler?, Walter В. Hoeh* & Margaret Mulvey? ABSTRACT Freshwater mussels in the genus Pleurobema (Bivalvia: Unionidae) from the eastern Gulf drainages of Florida, Georgia, and Alabama are variable conchologically, and several taxonomic names have been applied to shell phenotypes. Clarification of the species status of these mus- sels is important because they are recognized as imperiled throughout their range, mainly due to habitat destruction. Allozyme electrophoresis and restriction fragment length polymorphisms (RFLPs) of the nuclear ribosomal array showed little genetic differentiation among samples rep- resenting the phenotypes designated P. pyriforme and P. bulbosum. Partial DNA sequences of the mitochondrial cytochrome с oxidase subunit 1 distinguished P. reclusum from P. pyriforme and P. bulbosum. Genetic differentiation was observed between other Pleurobema species, specifically P. strodeanum, Р. clava, and P. sintoxia. Genetic evidence indicates that the eastern Gulf Coast Pleurobema (P. pyriforme and P. bulbosum) are a single species, P. pyriforme, with variable shell characteristics. Pleurobema reclusum is different genetically and may represent a distinct valid species. Pleurobema pyriforme is a federally endangered species. Although we hesitate to call P reclusum a distinct species because we were able to sequence DNA from only one individual, it is distinct genetically from P. pyriforme of the Apalachicola-Chattahoochee-Flint rivers basin and should merit conservation status. Key words: Pleurobema, Unionidae, RFLP analysis, allozymes, DNA sequencing, conserva- tion, endangered species. INTRODUCTION Freshwater unionacean mussels are among the most endangered fauna in North America (Bogan, 1993; Williams et al., 1993; Stolzenburg, 1995; Mulvey et al., 1997; Neves et al., 1997). North American unionids have a life cycle that requires a host (typically fishes) for development and metamorphosis of the glochidia, the unionid larval stage. When host species decline, the mussels that depend on them decline as well (Bogan, 1993). Mussel habitat has been destroyed by siltation of rivers, stream channelization, in- stream gravel mining, dredging, and damming and polluted from paper mills and chemical factories, untreated municipal effluent, and non-point source pollution (Bogan, 1993; Neves et al., 1997). Freshwater mussels have been commercially harvested for the pearl button industry in the early twentieth century (Bogan, 1993) and more recently for the Indo- Pacific pearl trade, for which their shells are ground into beads to form a nucleus for cul- tured pearls (Williams et al., 1993). Intro- duced species, including the Asian clam (Cor- bicula fluminea) and the zebra mussel (Dreissena polymorpha), compete with the native bivalves for space and food or, in the case of zebra mussels, actually use the native mussels as substratum (Bogan, 1993). World- wide, freshwater bivalves face similar pres- sures from humankind (Bogan, 1993). Many freshwater mussel species are cur- rently protected under the Endangered Species Act (ESA) of 1973 (amended in 1978). Conservation of species under the ESA requires that species be recognized as distinct from other such groupings. However, species discriminations often depend upon characters that are phenotypically variable. Information regarding reproductive isolation or underlying genetic distinctiveness may not be available for most molluscan taxa. Taxono- ¿University of New Orleans Department of Biological Sciences New Orleans, Louisiana 70148, U.S.A.; kkandl @ uno.edu Southwest Missouri State University Department of Biology 901 South National Avenue Springfield, Missouri 65804-0095, U.S.A. “United States Fish and Wildlife Service Asheville Field Office 160 Zillicoa St. Asheville, North Carolina 28801, U.S.A. Kent State University Department of Biological Sciences Kent, Ohio 44242-0001, U.S.A. Virginia Institute of Marine Science Department of Environmental Science The College of William and Mary Gloucester Point, VA 23602-1346, U.S.A. 88 KANDL ET AL. mists may lump phenotypic variants under a single species name, or conversely, may apply a specific name to each variant. For efforts towards conservation of biodiversity, failure to recognize cryptic species or prolifer- ation of taxonomic names can create man- agement problems. The ESA protects species of most organ- isms and populations of vertebrates, thus the recognition of valid species is critical to the implementation of the ESA. Several species concepts are currently in use, each differing from the others in their specific criteria (Roe & Lydeard, 1998). Two “universal” criteria of species concepts are (1) reproductive isola- tion and (2) distinct evolutionary lineages (Sites & Crandall, 1997). Populations of inver- tebrates are not legally protected under the ESA. In an effort to gain recognition, if not pro- tective status for populations of invertebrates and other organisms, the concept of evolu- tionary significant units (ESUs) has been em- ployed (Waples, 1991; Moritz, 1994). An ESU is a “population (or group of populations) that: (1) is substantially reproductively isolated from other conspecific units, and (2) repre- sents an important component in the evolu- tionary legacy of the species” (Waples, 1991; Bowen, 1998; Roe & Lydeard, 1998). Roe & Lydeard (1998) argue that the ESU concept overlaps considerably with the phylogenetic species concept, which suggests that a species is “the smallest aggregation of popu- lations or lineages diagnosable by a unique combination of character states”. Thus, some populations that are considered to be ESUs may reasonably be called species instead. For example, certain populations of Potamilus inflatus could be considered two distinct species because the populations are topolog- ically disjunct on a cladogram and reproduc- tively isolated, occurring in two different drainages (the Amite and the Black Warrior drainages) and therefore represent “phyloge- netically and evolutionarily distinct entities” (Roe & Lydeard 1998). The genus Pleurobema currently com- prises at least 32 species (Turgeon et al., 1998; Williams et al., 1993), although Simp- son (1914) recognized 76 species. At least 22 (71%) of the currently recognized species of Pleurobema are designated as threatened or endangered by Williams et al. (1993). As few as two and at least as many as nine nominal species of Pleurobema have been recognized from the Gulf drainages of Florida, Georgia, and Alabama (Clench & Turner, 1956; Simp- son, 1914). Historical records show that Pleu- robema was widespread and locally abundant throughout the Coastal Plain and Piedmont regions of the tri-state area (Brim-Box & Williams, 2000). Simpson (1914) recognized Pleurobema pyriforme (oval pigtoe), P. bulbosum (inflated clubshell), and P. reclusum (Florida pigtoe) based on morphological characteristics. These species, as well as Р modicum, were later synonomized under Р pyriforme by Clench 4 Turner (1956), who argued that vari- ation in shell characters did not reflect under- lying species differences. Some recent taxo- nomic treatments have again recognized P. bulbosum and P. reclusum as species distinct from P. pyriforme (Stansbery, 1976; Williams 8 Butler, 1994). Pleurobema pyriforme, how- ever, is the only species of Pleurobema cur- rently recognized by Turgeon et al. (1998) as occurring in northern Florida and southern Georgia from the Apalachicola River drainage to the Suwannee River drainage. Whether P. bulbosum and P. reclusum are distinct species is a critical conservation issue. These mussels are rare in their ranges and threat- ened by habitat destruction (Williams & Butler, 1994). Within the context of the Endangered Species Act, however, only taxonomically dis- tinct species of invertebrates may be consid- ered for conservation status. We examined specimens of Pleurobema from northern Florida, southern Georgia, and Alabama, including specimens identified as P. pyriforme, P. bulbosum, P. reclusum, and P. strodeanum (fuzzy pigtoe) based on morpho- logical and historical collection site data. Pleu- robema pyriforme occurs from the Econfina Creek drainage in west Florida to the Suwan- nee River system, although Clench & Turner (1956) recognized it as occurring only in the Apalachicola and Suwannee river systems, and Williams & Butler (1994) considered it to be an endemic of the Apalachicola River sys- tem. Pleurobema pyriforme is known from Econfina Creek drainage from 1970 and 1993 (USGS) records (J. D. Williams, personal communication to RSB). The type locality of Р pyriforme is near Columbus, Georgia (Simp- son, 1914). The range of P. reclusum includes the Ochlocknee and Suwannee river systems (Simpson, 1914; Williams & Butler, 1994), and its type locality is the Ochlocknee River in Leon County, Florida (Simpson, 1914). Stans- bery (1976) recognized P. bulbosum as a dis- tinct species from P. pyriforme. The historic range of Р bulbosum was the Flint and Oc- TAXONOMIC AMBIGUITY AND CONSERVATION 89 mulgee river systems of Georgia (Simpson, 1914), although the type locality, Macon, Georgia, on the latter river, is likely incorrect (Clench 8 Turner, 1956; J. Brim-Box, personal communication to KLK). The type locality con- sidered valid is Spring Creek, Decatur County, Georgia, which is a tributary to the Flint River and part of the Apalachicola River drainage (J. Brim-Box, personal communica- tion to KLK). Pleurobema strodeanum is con- sidered a distinct species (Williams & Butler, 1994). Its range extends from the Escambia River to the Choctawhatchee River system (Clench & Turner, 1956), and the type locality is the Escambia River in Florida (Simpson, 1914). In Florida, P. strodeanum is considered a threatened species (Williams & Butler, 1994). The taxonomy of freshwater mussels has been largely based on conchological charac- ters (Mulvey et al., 1997). Reliance on con- chological characteristics for taxonomic reso- lution is often problematic, because shell and nacre color, shell size, and shell shape can vary with environmental conditions and geo- graphic location (Stiven & Alderman, 1992; Mulvey et al., 1997). Molecular genetic tech- niques can provide additional information to better discriminate among species and to de- termine relationships among species which are unclear based on morphological charac- teristics alone (Davis, 1994; Hoeh, 1990; Hoeh et al., 1995; Mulvey et al., 1998). The objectives of this study were to assess the genetic distinctiveness of populations of Pleurobema in order to clarify the species status of eastern Gulf Pleurobema (P. pyri- forme, P. bulbosum, and P. reclusum). Al- lozyme electrophoresis, nuclear RFLPs, and mitochondrial DNA sequencing provided ge- netic characteristics and were used to evalu- ate relationships. To facilitate interpretation of the data, we compared our results to those of other studies that generated comparable ge- netic data for other freshwater mussel taxa. METHODS Twenty-eight specimens of Pleurobema from eleven northern Florida, southern Geor- gia, and southern Alabama populations were used for DNA analysis in this study (Table 1, Fig. 1). Pleurobema strodeanum, P. sintoxia, and P. clava were included to evaluate inter- specific genetic differences among Pleu- robema. Other genera from the subfamily Am- bleminae (sensu Lydeard et al., 1996), to which Pleurobema belongs, were selected for outgroups; these were Uniomerus carolini- ana, Elliptio sp., Quadrula apiculata, and Vil- losa lienosa. Based on a phylogenetic analy- sis of the 16S rRNA gene, Uniomerus and Elliptio are closely related to Pleurobema; Quadrula and Villosa are more distantly re- lated (Lydeard et al., 1996). Allozyme electrophoresis was performed on adductor muscle tissue from all specimens that were frozen (Table 1), and the following loci were scored: a-glycerophosphate dehy- drogenase (aGpd; E. С. 1.1.1.8), fluorescent esterase (F-Est; E. C. 3.1.1.1), glyceralde- hyde-3-phosphate dehydrogenase (Gapd; E. C. 1.2.1.12), glucose-phosphate isomerase 1 (Gpi-1; E. C. 5.3.1.9), lactate dehydrogenase (Ldh; Е. С. 1.1.1.27), malate dehydrogenase 1 and 2 (Mdh-1, Mdh-2; E. C. 1.1.1.37), malic enzyme (Me; E. C. 1.1.1.40), peptidase using leucylalanine and phenylanylproline as sub- strates (la-Pep, pp-Pep; E. C. 3.4.-.-), man- nose-6-phosphate isomerase (Mpi; Е. С. 5.3.1.8), nucleoside phosphorylase (Np; E. C. 2.4.2.1), and phosphoglucomutase-2 (Pgm-2, E. C. 5.4.2.2). Loci were analyzed on the fol- lowing buffer systems: Gapd, Mpi, and Pgm-2 on Tris-citrate, pH 8.0 buffer (Selander et al., 1971), Np and Lah on Tris-citrate-EDTA, pH 7.1 buffer (Ayala et al., 1972); F-est, Me, Gpi, and «Gpd on Tris-EDTA-borate, pH 8.0 buffer (Selander et al., 1971); la-Pep and pp-Pep on lithium-hydroxide buffer, pH 8.1 (Selander et al., 1971); Mdh-1 and Mdh-2 on amine-citrate, pH 6.1 buffer (Clayton & Tretiak, 1972). The most common allele was designated 100, and other alleles were assigned according to their mobilities relative to the 100 allele and the ori- gin. BIOSYS-1 (Swofford & Selander, 1981) was used to determine allele frequencies and to generate a UPGMA phenogram based on Nei’s (1972) unbiased genetic distance. Genomic DNA, for RFLP analysis and DNA sequencing, was isolated following Liu et al. (1996). A 2.2 kilobase nuclear DNA fragment of the ribosomal region (partial 18S rRNA, ITS1, 5.8S rRNA, ITS2, and partial 28S rRNA) was amplified via polymerase chain reaction (PCR) using the primers, mussel 18S and mussel D3 (Mulvey et al. 1998). The ribosomal region DNA was digested with the following ten restriction enzymes: Рае |, Hae ll, Hha Ill, Hinf\, Mbo |, Mbo Il, Mse |, Msp |, Rsa |, and Taq |. Digests were run on 3% agarose gels with molecular weight markers and visualized with ethidium bromide. Polaroid photographs 90 KANDL ET AL. TABLE 1. Species and populations examined using RFLP (R), allozyme electrophoresis (A), and DNA sequencing of the СО! gene ($). The number of individuals examined using DNA analyses, population iden- tification letter (corresponding to Fig. 1 and Table 4), and location are also given. Taxa Study No. Ind. Pop. ld. Drainage River County, State P. bulbosum RAS 3 A Chipola Dry Creek Jackson, Florida P. reclusum RAS 2 B Suwannee New River Union, Florida Р pyriforme RA 1 C Chipola Chipola River Calhoun, Florida P. pyriforme RA 2 D Flint Kinchafoonee Webster, Georgia Creek P. pyriforme RAS 2 E Flint Kinchafoonee Webster, Georgia Creek P. pyriforme RAS 2 Е Flint Chickasawhat- Terrell, Georgia chee R. P. pyriforme RAS 2 G Chipola Baker Creek Jackson, Florida P. pyriforme RAS 5 H Econfina Econfina River Bay, Florida Psp: RAS 1 H Econfina Econfina River Bay, Florida PSP. RAS 3 | Chattahoochee Sawhatchee River Early, Georgia P. clava 5 1 Ohio Little Darby Creek Madison, Ohio P. strodeanum RAS 3 J Escambia Conecuh River Pike, Alabama P. strodeanum RA 2 K Escambia Escambia River Escambia, Alabama P. sintoxia RS 3 Mississippi Spring River unknown, Kansas P. sintoxia RS 2 Mississippi Mississippi River unknown, Kansas Amblema plicata S 1 Genbank Elliptio sp. RAS 2 Chattahoochee Sawhatchee River Early, Georgia Elliptio crassidens S 1 Apalachicola Apalachicola Gadsen, Florida River Fusconaia flava S 1 Genbank Quadrula apicu- S 1 Alabama Little River Baldwin-Monroe, lata Alabama Uniomerus car- RA 1 Altamaha Altamaha River Long, Georgia oliniana Villosa lienosa A 2 Kissimmee Lake Arbuckle Polk, Florida were taken and fragment lengths were deter- mined by comparison with markers from the photographs. Restriction sites were mapped onto the nuclear ribosomal region by compar- ing fragment patterns to a related species (Е!- liptio complanata), with known restriction sites (Liu & Mulvey, unpublished data). For frag- ment sizes that differed from E. complanata, probable cutting sites were inferred. For all study mussels, there was always a partial match of fragment sizes to those of E. com- planata. Both the presence/absence of frag- ments and the restriction sites were analyzed by maximum parsimony (PAUP* 4.0 Beta ver- sion; Swofford, 1998) to examine the evolu- tionary relationships among taxa. A heuristic search using the MULPARS option was used to find the most parsimonious trees. A boot- strap analysis was performed using 2000 bootstrap replicates and a heuristic search to obtain bootstrap values on a 50% majority rule consensus tree. A total evidence approach, in which al- lozyme and RFLP data were combined, was also utilized. For this analysis, allozyme data were coded as character states with loci as characters. Specimens exhibiting allozyme polymorphisms were coded as having multi- ple character states. For example, a speci- men having two different alleles at a particular locus might be coded as having a character state of 1,2. A 710 base pair segment of the mitochon- drial cytochrome c oxidase subunit 1 gene (CO1) was amplified via PCR using primers, COIL 1490: 5'-GGT CAA CAA ATC ATA AAG АТА TTG G-3', and COIL 2198: 5'-TAA ACT TCA GGG TGA CCAAAAAAT CA-3’) (Folmer et al., 1994), but only 347 base pairs of this sequence were included in the analysis. DNA amplifications followed protocols established in Hershler et al. (1999). Amplification prod- ucts were purified with Microcon™-100 fil- ters. Purification products were used as tem- plate for automated sequencing following Applied Biosystem protocols. Sequences were aligned using Sequencher™ software and by visual examination. Relationships 91 TAXONOMIC AMBIGUITY AND CONSERVATION | эаег ul papinold эле suone90] sainads JO sjiejap Jayuny JOAIY yNn9au0) ‘шпиверод$ y (У) any elqueos3 ‘шпиеэрол$ y (Г) 49819 seyojeymes “ds y (|) IOAIH еициоэз “ds y pue эциои/а y (H) 49919 sayeg ‘auuouAd y (9) 49819 ээчэзецмезехэ!ц9 ‘euuoyuAd y (4) 48819 aauoojeyouy ‘эииоила y (3) ‘48819 ээцооецоим ‘euuoquAd y (а) UEAIY ejodiyo BunoyuAd y (9) ‘зэл MAN “unsn/9al а (a) ‘48819 Ала ‘wnsoging 4 (Vv) ‘ерно|+4 pue ‘eiBioay ‘ешеадем шо says U0I98/|09 euusqolnay JO зиоцеэол “| ‘59+ LC EEE ЭП STIL 90 1 0 MINE JIUUEMNS 2131020) $ Е S V 3 вигедету 32 KANDLET AL. among samples were examined using maxi- mum parsimony analysis (PAUP* 4.0 Beta version; Swofford, 1998). A heuristic search was used to find the most parsimonious trees and a bootstrap analysis was performed using 1000 replicates. RESULTS Ten of the eleven allozyme loci were poly- morphic across all taxa; only Gapd was monomorphic across all taxa. Eight loci were monomorphic for the eastern Gulf Pleu- robema (P. pyriforme, P. bulbosum, and P. reclusum) but were polymorphic across other taxa (Table 2). All allele frequencies are based on sample sizes of less than eight. Thus, this allozyme study can be used in a qualitative fashion to compare the presence/absence of alleles in populations, but should not be used to compare allele frequencies among popula- tions. The phenogram based on allozyme data contains a cluster consisting of speci- mens designated as P. pyriforme, P. bulbo- sum, and P. reclusum based on conchological traits (Fig. 2). The genetic distances (Nei, 1972) between pairs of eastern Gulf Pleu- robema ranged from 0 to 0.058 (Fig. 2, Table 3). The genetic distance between Р stro- deanum and taxa in the eastern Gulf Pleu- robema cluster ranged from 0.117 to 0.227, and the genetic distance between the two populations of P. strodeanum was 0.114. The average pair-wise genetic distances between the eastern Gulf Pleurobema taxa and the outgroups, Uniomerus caroliniana and Villosa lienosa, were 0.989 + 0.016 and 1.206 + 0.016, respectively. Analysis of fragment presence/absence and restriction site presence/absence in the ribosomal array using PAUP produced similar results, therefore only restriction site data will be presented. The ten restriction enzymes produced 94 cutting sites (Fig. 3) and nine haplotypes (Table 4). Twenty-two out of 23 samples that had been identified as P. pyri- forme, P. bulbosum, P. reclusum, or P. sp. were the same haplotype (АААААААААА); only one P. pyriforme from Baker Creek had a different haplotype (AAAAAABAAA). Maximum parsimony of RFLP restriction sites produced 14 equally parsimonious trees when characters were weighted equally (TL = 72, Cl = 0.931, RI = 0.762). A 50% majority rule consensus tree is shown in Figure 4. Eastern Gulf Pleurobema, including speci- mens designated as P. pyriforme, P. bulbo- sum, P. reclusum, and P. sp., form a single clade. Pleurobema strodeanum from the Es- cambia River and Conecuh River form a dis- tinct clade, and this clade is the sister taxon to the P. pyriforme clade. The combined data from restriction site and allozyme elec- trophoresis yielded a tree that was nearly identical to that in Figure 2. Maximum parsimony analysis of the CO1 mitochondrial gene sequences produced a single most parsimonious tree when all char- acters were weighted equally (TL = 148, Cl = 0.730, RI = 0.523) (Fig. 5). The sample identi- fied as P reclusum from the Suwannee River is the sister taxon to other Pleurobema speci- mens from the eastern Gulf Coast identified as P. pyriforme, P. bulbosum, and P sp. The sequence divergence between P. reclusum and the other eastern Gulf Coast specimens was 2.31%, whereas the sequence diver- gence among the other eastern Gulf Coast specimens was less than 0.2% over the 347 bp sequence analyzed. Pleurobema stro- deanum + P. clava formed the sister clade to all of the eastern Gulf Pleurobema; the aver- age sequence divergence between these species was 5.4%. Interspecific sequence di- vergence values were 5.76% for P. sintoxia vs. P. strodeanum, 5.19% for P. pyriforme vs. P. sintoxia, 5.76% for P. pyriforme vs. P. clava, 6.34% for P. clava vs. P. sintoxia, and 6.05% for P. clava vs. P. strodeanum. The eastern Gulf Pleurobema clade was most closely re- lated to the other Pleurobema species in this analysis, including P. clava, the type species of the genus Pleurobema. The Pleurobema clade was most closely related to Fusconaia flava, the type species of the genus Fus- conaia. Fusconaia is considered to be the sis- ter genus to Pleurobema (Davis & Fuller, 1981). DISCUSSION Specimens referred to as Pleurobema pyri- forme, P. reclusum, and P. bulbosum (eastern Gulf Pleurobema) show substantial morpho- logical variation. In particular, P. reclusum is morphologically distinct from P. pyriforme and P. bulbosum in that P. reclusum is not quite so inflated and is more plicate having two promi- nent ridges (Simpson, 1914; Williams & Butler, 1994). Genetic evidence from DNA sequenc- ing of the CO1 mitochondrial gene supports the hypothesis that P reclusumis distinct from the other eastern Gulf Pleurobema. Sequence TAXONOMIC AMBIGUITY AND CONSERVATION 93 TABLE 2. Allele frequencies determined by allozyme electrophoresis. Population Id refers to the letter of the population in Fig. 1. No. Ind. refers to the number of individuals that were used in this allozyme study. The following abbreviations are used for taxa: Pp = P. pyriforme, Pb = P. bulbosum, Pr = P. reclusum, Psp = P. sp., Ps = Р strodeanum, Uc = Uniomerus caroliniana, VI = Villosa lienosa. Locus and Allele Pop ld. H G D E F A B H | K J No. Ind. 2 if 6 5 2 4 2 2 7 2 4 2 2 Taxoh “Pp Rp "Pp" "Pp “Pp Pb Pr 0.08 1.00 1.00 0.92 0.90 1.00 1.00 1.00 1.00 1.00 1.00 95 1.00 1.00 80 0.10 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1:00) 1:00’ 1:00 1.00) 1:00’ 1.00) 1.007 1.00 1.00 1.00 COMM OO DOI COMME 100" 1-00) 1:00 TOOMMEOOI DOM OO 1:00) 1:00’ 1:00’ 1.00 1.00 OO OUI OO DOM OO 1:00) 1:00) 1:00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 96 1.00 0.50 84 0.50 1/005) 1200) 1.00 O0 1200) 1.00 100 1.00 95 1.00 84 1.00 1.00 1.00 1.00 0.50 0.50 0.17 0:17 0.50 0.50 1.00 0.83 0.83 1.00 1.00 0.50 0.50 1.00 1.00 0.88 90 0.12 0.33 112 0.20 1:00" 1.00, 0:67 0.80 0:50 0:88 1.00. 1.00. 0:29 0:88, 1.00) 1.00 81 1.00 0.50 0.12 0.71 0.12 divergence between P. reclusum and P. pyri- forme was 2.31% for the 347 bp region ana- lyzed. This level of divergence is midway be- tween the levels of divergence within the P pyriforme clade (all less than 0.29%) and the levels of divergence for pairwise comparisons among distinct Pleurobema species (5.19%- 6.34%). This level of sequence divergence is not unexpected, given the geographic isola- tion of the Apalachicola-Chattahoochee-Flint rivers and Suwannee River drainage popula- tions. 94 KANDL ET AL. . pyriforme - Econfina sp. - Econfina . pyriforme - Baker . pyriforme - Kinchafoonee . pyriforme - Kinchafoonee . pyriforme - Chickasawhatchee . bulbosum - Chipola . reclusum - Suwannee . Sp. - Sawhatchee . strodeanum - Escambia ach) pack a Sac a A a as) . strodeanum - Conecuh Uniomerus - Altamaha Villosa - Kinchafoonee А ГР ВЕ 1.0 0.5 0.4 0.3 0.2 0.1 0.0 EE + + 4 Genetic Distance (Nei, 1972) FIG. 2. UPGMA phenogram based on allozyme data. Branch lengths represent Neïs (1972) genetic dis- tance. TABLE 3. Comparison of genetic distances based on allozyme studies (Nei, 1972) + standard deviations among populations within a species and species within a genus of freshwater mussels. No. Pop. No. Taxon or species Loci Populations within a species Anodonta cataracta 5 NA Elliptio complanata 11 8 Lampsilis cariosa 3 11 Lampsilis radiata 5 7 Leptodea ochracea 2 11 Pleurobema pyriforme 9 13 Species within a genus Amblema 3 14 Anodonta 3 14 Elliptio 7 14 Lampsilis 6 14 Megalonaias 2 14 Pleurobema 2 13 Uniomerus 3 14 Allozyme analyses and RFLP data do not differentiate among any of the eastern Gulf Pleurobema populations. The average among-population genetic distance (Nei, 1972) based on allozymes for eastern Gulf Pleurobema was 0.031 with a maximum value of 0.058. This value is well within the range of among-population genetic distances for other Genetic Distance Reference 0.034 + 0.038 Davis, 1994 0.065 + 0.039 Davis et al., 1981; Davis, 1994 0.071 + 0.027 Stiven & Alderman, 1992 0.018 + 0.010 Kat & Davis, 1984 0.018 Stiven & Alderman, 1992 0.031 + 0.029 this study 0.219 + 0.025 Mulvey et al., 1997 0.457 = 0073 Kat, 1983a 0.210 + 0.017 Davis et al., 1981; Davis, 1984 0.609 + 0.478 Kat, 1983b all < 0.100 Mulvey et al., 1997 0.185 + 0.045 this study 0.308 + 0.165 Davis et al., 1981; Davis 1994 unionid species (Table 3). In addition, the phy- logenetic trees from the RFLP and sequence data show support for a single clade consist- ing of P. pyriforme, P. bulbosum, and P. reclusum. This clade is represented by the large polytomy in Figure 4. Allozyme elec- trophoresis and RFLP analysis of a nuclear gene are techniques that are less likely to dis- TAXONOMIC AMBIGUITY AND CONSERVATION 95 Die ЕЕ =a A E A AAA AN ] Fe Du A He E Re A [| В io М И — A m C a Bi Kerne EA Hha MM — — ee] pa À Be SENA El ESC [MEA Е] [| = Aero A ih) (X a E ЕЕ CE E B EXA AA Fe oe a Win = = AAA A AST A eel a Е SEPA C a An ae CET Mbo Il = = AAA een] [+] a] C ESS Bern El A Msc | ЕК Ged ЕЕ eee |" © SRE Bene В С Bu [PERE] = D Msp! I = er TEE EEN E | A NC | [E] E El A Rsa I aaa pra] SRE) € = — | E] fe ee EEE) A TagT =o M mil: ES HERE CE ES en Be O | CA в RE D 0 500 1000 1500 2000 FIG. 3. Cleavage maps from the 10 enzymes used to cut the 2.2 kb r RNA gene. The restriction enzymes producing the fragments are listed on the left side; capital letters that designate restriction digest patterns are listed on the right. The fragment sizes for each cleavage map are given in Table 5 in the order in which they appear in the figure. 96 KANDL ET AL. TABLE 4. RFLP haplotype counts in the populations studied. Letters refer to the populations of Pleurobema listed in Table 1 and in Fig. 1. Psin-Sp = P. sintoxia from the Spring River, Psin-Mi = P. sintoxia from the Mississippi River, Esp = Elliptio sp. and Uc = Uniomerus caroliniana. Population Haplotype A TB. GD EME @ 1 J К Psin-Sp Psin-Mi Esp Uc AAAAAAAAAA A IR ОР 3 == = = = = AAAAAABAAA 1 a = = AAAAAADAAA AUS IS 5 Y 2 > = = = AAABAADAAA 2. = = = = = AABAAAAAAA 3 = = — AABAAAAAAD 1 = = ABBAAAAAAD 1 = = AAAAAABAAB = 2 = ACCCCCCCCC = = 1 TABLE 5. RFLP fragment sizes for each cleavage map given in Fig. 3. Restriction Restriction Enzyme Digest Pattern Restriction Fragment Size Dde | A 343 316 141 354 272 98 279 347 Hae Il A 788 524 27 237 119 218 176 61 B 324 464 524 27 237 119 218 176 61 C 300 57 164 81 186 551 237 241 272 61 Hha Ш A 205 35 410 589 495 248 168 B 205 35 610 389 495 248 168 C 205 645 389 461 34 248 168 Hinf | A 951 229 44 154 68 120 357 46 181 B 951 273 154 68 120 357 46 181 C 951 -229 198 68. 120 357 227 Mbo | A 123 194 257 95 447 190 747 97 C 123 161 290 95 447 190 747 97 Mbo Il A 692 353 157 2911696 Е 830) 215. 157.292. 156500 Mse | A 197 216 303 368 57 1009 B 197 216 303 425 1009 C 197 203 316 384 41 1009 D 197 216 303 355 70 1009 Msp | A 96 149 84 267 34 143 10 777 590 C 96 149 84 267 34 143 10 857 510 Rsa | A 12.327 61 607 “283 174 47 381 258 С 12 318. 70 700 4190 221 381 258 Tag | A 90 133 162 79 13 184 77 594 489 329 B 90 133 162 179 44 79 197 77 594 489 106 C 90 133 162 179 44 79 38 159 77 389 450 200 150 D 90 133 162 191 32 79 18 179.77 594 1489 tinguish among closely related taxa than is DNA sequencing of the CO1 gene, and nei- ther the allozyme data nor the RFLP data pro- vide sufficient resolution to distinguish these taxa (Hillis et al., 1996). All three genetic techniques clearly differen- tiate P. strodeanum, P. sintoxia, and P. clava from the eastern Gulf Pleurobema. Analysis of allozyme data reveal distances for P. stro- deanum vs. P. pyriforme that are all greater than 0.117. Although there is no absolute ge- netic distance value that separates popula- tions within a species or species within a genus, comparisons with genetic distances of taxa that are widely accepted to be differenti- ated at the population or species level may be useful in identifying the appropriate level of taxonomic classification (Davis, 1994). Both RFLP analyses and DNA sequence data sup- port the notion that P. strodeanum is geneti- cally distinct from P. pyriforme as well. Both RFLP and sequence data analyses group the two or three populations of P. sintoxia and sep- arate P. sintoxia from P. strodeanum, P. clava, TAXONOMIC AMBIGUITY AND CONSERVATION 97 P. bulbosum - Chipola P. reclusum - Suwannee P. pyriforme - Kinchafoonee P. pyriforme - Kinchafoonee 37 P. pyriforme - Chickasawhatchee P. pyriforme - Chipola 42 P. pyriforme - Econfina 78 P. sp. - Sawhatchee auofiadd “y DU9qOAN9]4 P. sp. - Econfina P. pyriforme - Baker P. strodeanum - Escambia 61 31 P. strodeanum - Conecuh 18 P. sintoxia - Spring P. sintoxia - Mississippi = Elliptio sp. Uniomerus FIG. 4. 50% majority rule consensus tree (from 14 equally parsimonious trees), based on restriction site data for Pleurobema taxa, Uniomerus caroliniana, and Elliptio sp. Numbers are bootstrap values after 1000 repli- cates. and the eastern Gulf Pleurobema. The genus Pleurobema is monophyletic in both the RFLP and DNA analyses although bootstrap values indicate relatively poor support for this clade (Figs. 4, 5). Two main conclusions arise from these data sets. The first is that allozyme elec- trophoresis, RFLP analyses, and DNA se- quencing did not differentiate specimens identified as P. bulbosum from P. pyriforme, although these same techniques certainly dis- tinguish currently recognized species of Pleu- robema. The range of P. bulbosum is the Flint River, which is part of the Apalachicola River drainage. The range of P. pyriforme also in- cludes the Apalachicola River drainage (Williams & Butler, 1994). Thus, specimens identified as P. pyriforme or P. bulbosum may represent intraspecific morphological varia- tion that occurs in different rivers within the same drainage or they may represent distinct species. Pleurobema bulbosum and P. pyri- forme can be distinct species despite the low levels of genetic divergence (Sites & Crandall, 1997), but at this time and based on these ge- netic data, we can not recommend that they be considered distinct species. Pleurobema reclusum is genetically distinct 98 KANDL ET AL. 54 P. strodeanum - Escambia P. clava - Ohio P. pyriforme - Chickasawhatchee 56 P. sp. - Sawhatchee 80 | P. pyriforme - Chipola P. pyriforme - Econfina au1ofiudd q ошадолт A 97 P. pyriforme- Kinchafoonee 38 P. bulbosum - Chipola P. reclusum - Suwannee 50 65 | P. sintoxia - Mississippi 99 P. sintoxia - Spring 82 P. sintoxia - Spring F. flava E. crassidens A. plicata O. apiculata FIG. 5. The single most parsimonious phylogram based on a 347 bp sequence of the mitochondrial gene, COI. Numbers are bootstrap values, after 2000 bootstrap replicates. from other eastern Gulf Pleurobema at the CO1 mitochondrial gene using DNA analysis. Regrettably, we had a tissue sample from only a single individual identified as P. reclusum from the Suwannee River. We hesitate to say that this one specimen represents a distinct species of Pleurobema, P. reclusum. On the other hand, because of these genetic differ- ences, we hesitate to say that this individual simply represents morphological and genetic variation present in P. pyriforme. Pleurobema reclusum is confined to the Ochlocknee and Suwannee River drainages, and it is unlikely that gene exchange occurs with Р pyriforme from the Apalachicola drainage; thus, we at- tribute the CO1 divergence to geographic iso- lation, and perhaps reproductive isolation as well. Additional samples are needed to deter- mine whether this morphological and genetic variation is indicative of population-level dif- ferences or whether it is indicative of a nascent species-level divergence. Further studies to differentiate between these possi- bilities should include samples from the Ochlocknee River. Several attempts on our part to include specimens from the Ochlock- nee were unsuccessful. These samples are important because the Ochlocknee River, be- tween the Apalachicola drainage where P. pyriforme is found and the Suwannee River, is an area in which P. pyriforme and P. reclusum may co-occur. The Suwannee River drainage may repre- sent an area of endemism for freshwater mus- sels and other animals. Lydeard et al. (in press) showed that Quincuncina infucata in TAXONOMIC AMBIGUITY AND CONSERVATION 99 the Suwannee River is substantially geneti- cally different from other Q. infucata and sug- gested that populations in the Suwannee River be raised to the species level. Likewise, Hoeh et al. (1995) found a distinct clade of Ut- terbackia peggyae in the drainages of north- western peninsular Florida, including the Suwannee River drainage. Populations of al- ligator snapping turtles from the Suwannee River are morphologically and genetically dis- tinct from other such populations, and this river represents the deepest evolutionary break in the phylogeny of alligator snapping turtles (Roman et al., 1999). One explanation for this region of endemism is that this area was inundated and then isolated from the sur- rounding region at least two times during the Cenozoic due to sea level changes (Hoeh et al., 1995). These vicariance events likely re- sulted in diversification of the freshwater fauna of the northern peninsula of Florida (Hoeh et al., 1995). Pleurobema in the eastern Gulf are in need of conservation (Williams et al., 1993; Williams & Butler, 1994). Clench & Turner (1956) noted that P. pyriforme (synonomized with Р reclusum and P. bulbosum in their study) was “relatively rare and perhaps only locally abundant”. In the 40 years since Clench & Turner (1956), destruction of fresh- water mussel habitat has continued at an ac- celerated pace. Stansbery (1971) noted that P. pyriforme was becoming less abundant in its range, and five years later it was desig- nated as threatened in Alabama (Stansbery, 1976). Williams et al. (1993) designated P pyriforme as endangered, and in 1998 “feder- ally endangered” status was proposed for Р pyriforme (U.S. Fish & Wildlife Service, 1998). Although it is equivocal whether Pleurobema in the Suwannee River drainage represent a valid species, we found a distinct genetic vari- ant at the CO1 locus in a specimen from this region. There is also evidence that popula- tions of several animals, including freshwater mussels, are distinct in the Suwannee River. Thus, we urge wildlife managers to treat the Pleurobema that occur in the Suwannee River drainage as a separate entity for conservation purposes. ACKNOWLEDGMENTS We wish to thank the following people and organizations for providing us with specimens of Pleurobema: J. Brim-Box, J. Williams, and H. Blaylock-Herod, BRD-USGS in Gaines- ville, Florida; T. King and M. Smith, BRD- USGS in Leetown, West Virginia; C. Lydeard, University of Alabama; A. Bogan, North Car- olina State Museum of Natural Sciences; and T. Watters, Ohio State University. J. Williams, E. Keferl, and J. Brim-Box painstakingly gave advice and information concerning collection locations, shell identification, and explana- tions for aberrant data. The comments of C. Lydeard and R. Dillon significantly improved this manuscript. C. Guy helped prepare the map. This work was supported by contract DE-FC09-96SR18546 between U.S. Depart- ment of Energy and the University of Geor- gia’s Savannah River Ecology Laboratory, and the U.S. Fish and Wildlife Service, Asheville, North Carolina. LITERATURE CITED AYALA, F.J., J.R. POWELL, M.L. TRACEY, C.A. MOURAO & S. 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Revised ms. accepted 1 September 2000 MALACOLOGIA, 2001, 43(1-2): 103-215 A SURVEY OF THE EUROPEAN VALVATIFORM HYDROBIID GENERA, WITH SPECIAL REFERENCE TO HAUFFENIA POLLONERA, 1898 (GASTROPODA: HYDROBIIDAE) Marco Bodon, Giuseppe Manganelli & Folco Giusti Dipartimento di Biologia Evolutiva, Universita di Siena, Via Mattioli 4, 1-53100 Siena, Italy; giustif @ unisi.it ABSTRACT All the taxa of genus and species groups introduced for the European valvatiform hydrobiids are analyzed, and if possible revised. All the type species have been redescribed on the basis of new anatomical studies or data in the literature when spirit specimens were not available. This enabled the following taxa to be rec- ognized as junior synonyms of Hauffenia Pollonera, 1898 (type species Horatia (Hauffenia) tellinii Pollonera, 1898): Erythropomatiana Radoman, 1978 (type species Valvata erythropoma- tia Hauffen, 1856); Lobaunia Haase, 1993 (type species: Lobaunia danubialis Haase, 1993); Neohoratia Schütt, 1961 (type species Valvata (?) subpiscinalis KuScer, 1932); and Vrania Rado- man, 1978 (type species Valvata wagneri Kuscer, 1928). Of the species currently assigned to Hauffenia, only a few from the eastern Alps and Balkans actually belong to it: H. erythropomatia (Hauffen, 1856); H. kerschneri (Zimmermann, 1930); H. media Bole, 1961; H. subcarinata Bole & Velkovrh, 1987; H. subpiscinalis ( Kuscer, 1932); H. tellinii (Pollonera, 1898) (with H. michleri Kuscer, 1932, as junior synonym); H. tovunica Rado- man, 1978; H. wagneri (KuScer, 1928); and H. wienerwaldensis Haase, 1992. Each is re- described. Many “Hauffenia” species from the Balkans remain of uncertain generic status due to the ab- sence, or incompleteness of anatomical data. However, Horatia (Hauffenia) raehlei Schutt, 1980, is placed in the genus Fissuria Boeters, 1981, on the basis of its anatomy. After anatomical study, most of the French “Hauffenia” have been assigned to /slamia Rado- man, 1973a, and tentatively to the following species: /. minuta (Draparnaud, 1805), /. globulina (Paladilhe, 1866), /. spirata (Bernasconi, 1985), and /. consolationis (Bernasconi, 1985). Anatomical study of the French “Horatia” exilis (Paladilhe, 1867) supports its allocation to a new genus: Heraultia, n. gen. The Spanish species currently assigned to “Horatia” or “Neohoratia” do not belong to these genera. Some of them, namely “N.” ateni (Boeters, 1969), “N.” globulus globulus (Bofill, 1909), and “N.” д. lagari (Altimira, 1960), belong to /slamia. Diagnoses and keys are provided for the European valvatiform hydrobiid genera and for the species of Hauffenia, together with tables summarising their taxonomic status and distribution. Key words: Gastropoda, Caenogastropoda, valvatiform Hydrobiidae, taxonomy, systematics, geographical distribution, Europe. INTRODUCTION The European fauna is particularly rich in valvatiform stygobiont hydrobiids (Radoman, 1983; Bole & Velkovrh, 1986). The first species discovered were assigned to the het- erobranch valvatid genus Valvata Muller, 1774, until the first hydrobiid genera were de- scribed. Since the 1950s, a plethora of new genera and new species have been intro- duced. This produced a puzzling taxonomic picture, with genera established on shell and/or few anatomical characters, frequent changes of rank of many supraspecific taxa, 103 and species (including extra-European; see Bole & Velkovrh, 1986) assigned to genera on the basis of non-diagnostic characters. In an attempt to reduce this taxonomic chaos, we set out to revise all the taxa related to one of the oldest established genera, Hauffenia Pol- lonera, 1898. Hauffenia was introduced by Pollonera (1898) as a subgenus of Horatia Bourguignat, 1887, for two new hydrobiids, collected by A. Tellini in the debris of the upper course of the Natisone River, Friuli, northeastern Italy: Ho- ratia tellinii and H. valvataeformis. Kuscer (1932, 1933a, b) ranked Hauffenia 104 BODON, MANGANELLI & GIUSTI as a distinct genus, but this was not substan- tiated until Bole (1970) studied the anatomy of H. tellinii and found that its genitalia were dif- ferent from that described by Radoman (1966) for the type species of Horatia. Bole (1970) listed three diagnostic characters of Hauffenia: (1) female genitalia with a small bursa copulatrix having short duct arising from the oviduct near where it enters the al- bumen gland (pallial oviduct), and a very small seminal receptacle arising from the renal oviduct where the oviduct loop ends; (2) male genitalia with a wide and flat penis; and (3) operculum with peg-like structure on its inner face. Boeters (1974) split Hauffenia into two sub- genera, Hauffenia (s. s.) and Neohoratia, the latter introduced by Schutt (1961a) as a sub- genus of Horatia for Valvata subpiscinalis Kuscer, 1932. Boeters (1974) regarded Neo- horatia as a subgenus of Hauffenia because H. subpiscinalis shares the genital characters of the species of Hauffenia (s. s.), but it has no peg-like structure on the inner face of the op- erculum. Oddly, Radoman (1978, 1983) overlooked Neohoratia but divided Hauffenia into two subgenera: Hauffenia (s. s.) and Vrania Rad- oman, 1978 (type species: Valvata wagneri Kuscer, 1928), the latter characterized only by an operculum with a thin edge and a small outgrowth on its inner face. Radoman (1978, 1983) also introduced a number of new gen- era from the Balkans, one of these, Erythro- pomatiana Radoman, 1978 (type species: Valvata erythropomatia Hauffen, 1856), had an organization of the female genitalia similar to that of Hauffenia. Subsequently, Bole & Velkovrh (1986) and Boeters (1988, 1998) ranked Neohoratia as a distinct genus. Boeters (1988, 1998) probably based this assumption on the fact that three Iberian species and one French species, sup- posed by him to be congeneric with N. sub- piscinalis, had differently organized female genitalia: the sac-like structures arising from the renal oviduct are two seminal receptacles and not a bursa copulatrix and a seminal re- ceptacle, as in Hauffenia. Recently, Bole (1993) re-examined the Slovenian taxa of this group, confirming Hauffenia, Erythropoma- tiana and Neohoratia as distinct genera, al- though they were characterized by rather in- significant anatomical characters. On the other hand, he treated Vrania as a junior syn- onym of Hauffenia, as proposed indepen- dently by Haase (1993). In this paper, we redescribe Hauffenia and define its relationships with nominal genera claimed to have similar anatomical organiza- tion (e.g., Erythropomatiana, Neohoratia, and Vrania). To do this, we first consider all the genera of the European valvatiform hydrobi- ids and redescribe their type species on the basis of literature and our own data. We also revise all the specific taxa assigned to these genera, claryifing which actually belong to Hauffenia and which belong to other genera. Each Hauffenia species is redescribed in the second section of the paper and a few mis- identified Hauffenia species are redescribed and discussed. MATERIAL AND METHODS Shells and live specimens were collected by sorting variable amounts of sediment from caves and springs. Unrelaxed material pre- served in 75% ethanol was studied by light microscopy (Wild M5A). Bodies were isolated after crushing the shells and were dissected using very fine, pointed watchmaker’s for- ceps. Images of the body and isolated parts of the genitalia were drawn using a Wild camera lucida. Radulae were obtained by dissecting out buccal bulbs and soaking them in KOH solution to remove soft tissue. They were washed in distilled water, mounted on copper blocks with electronconductive glue, sputter- coated with gold, and photographed using a Philips 505 SEM. Shells and shell fragments were photographed under light and scanning electron microscopes. The anatomical parts are disposed as in life position. So when the penis is described, terms such as left, right, ventral and dorsal correspond to the left, right, ventral and dorsal sides of the snail in life position. When a struc- ture belonging to an organ is described, its position is indicated in relation to the proximal origin of the organ (e.g., “at about 2/3 of penis length” means about 2/3 of the distance from the base to the apex of penis). The terms used in the description are those recently pro- posed by Hershler & Ponder (1998), except for the following (parentheses-terminology of Hershler & Ponder, 1998): last whorl (body whorl); protoconch malleated (pitted or wrin- kled); right (outer) or left (inner) side of penis; seminal groove (ventral channel); lateral wings of central tooth (lateral margins); lamel- lae (filaments). We prefer these terms be- cause they are in current use by European REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 105 authors or describe the aspect or function of the structure more exactly. In the description of the taxa, any reference to a structure will be omitted when its pres- ence/absence has been impossible to ascer- tain in an unequivocal manner (e.g., the hy- pobranchial gland) or when data in the literature is insufficient to allow their exact de- finition (e.g., the route of the penial duct). For identification of the different sac-like structures arising from the distal renal oviduct we adopted the following criteria, in the ab- sence of histological evidence: » the proximal seminal receptacle (PSR; sec- ond or RS2, according to Radoman, 1973b, 1983) leaves the oviduct far from the bursa and level with the end of the oviduct loop; e the distal seminal receptacle (DSR; first or RS1, according to Radoman, 1973b, 1983) arises very close to the point where the oviduct enters the albumen gland (pallial oviduct) but proximally with respect to the bursa copulatrix; it is obviously easy to identify when the bursa is present; when the bursa is absent, it can be identified by the fact it arises further from where the oviduct enters the albumen gland than any bursa, and consequently lies in a position corresponding to that of the DSR in species that also have a bursa; + the bursa copulatrix (BC) arises close to the point where the oviduct enters the albumen gland (pallial oviduct). When histological evidence exists, the bursa copulatrix (gametolytic gland) is a struc- ture that does not contain spermatozoa or contains few, non-oriented and partially di- gested spermatozoa (its contents are cen- trally located and never refringent), whereas the seminal receptacle/s is/are the structure/s that contains/contain spermatozoa oriented with their heads anchored to the cells of the wall (Thompson & Bebbington, 1969; Giusti & Selmi, 1985). For histological study, the penis was fixed in 75% ethanol, dehydrated, diaphanized with xylol, and mounted on microscope slides. The female genitalia (renal and pallial oviduct) were fixed in 75% ethanol, dehydrated, em- bedded in paraffin and cut serially; sections mounted on microscope slides were stained with haematoxylin-eosin. The material examined is listed as follows: locality name (municipality, district and/or country), UTM references, collector(s) and date (number of male and female specimens, shells, non-investigated specimens). Parsimony analysis was performed using a test version of PAUP (Version 4.0.b2 for Mac- intosh; Swofford, 1997). Tree search was per- formed using the heuristic search and the “collapse branches if minimum length is zero” options. The characters used for cladistic analysis were chosen on the list of Hershler & Ponder (1998). Only the type species of each genus was considered or, alternatively, that regarded to be the most closely related species, if the anatomy of the type species is not known. Much of the literature on the valvatiform hy- drobiids published after the fifties by S. Hadzisce and P. Radoman has questionable publication dates and conditions regarding the availability of the names. The main problem concerns the date of publication of two papers (one by HadZisce and the other by Radoman) on the hydrobiids of Lake Ohrid. Radoman (1963a: 69; b: 85-86) claimed that Hadzisce paper was published in 1959 and his paper in 1957: “Hadzisce’s paper, although antedated to August, 1956, was actually issued only in April, 1959 (date taken from the inventory of the Hydrobiological Institute at Ohrid), while my paper was issued in October, 1957 (date taken from the inventory of the Publishing De- partment of Kolorac National University ... in Beograd” (Radoman, 1963b: 86). Unfortu- nately, Radoman (1963a, b, 1973a, 1983) left room for confusion by continuing to cite the two papers with the putative year of publica- tion (1956) and not the true year of publication (1957 for his paper, 1959 for HadziSce’s paper). This caused misinterpretation by sub- sequent authors dealing with the taxa estab- lished in these papers (Bole & Velkovrh, 1986; Jovanovic, 1991; Kabat & Hershler, 1993; Dhora & Welter-Schultes, 1996). We accepted 1957 as the date of publica- tion of Radoman’s paper and 1959 as the date of publication of HadziSce’s paper. In the case of species described by both authors, Rado- man’s names therefore have priority over those of HadziSce. Consequently Ohrigocea (Karevia) prlitchevi Hadzisce, 1959, is a junior synonym of Pseudamnicola ornata Radoman, 1957, and Ohridohoratia (Ohridohauffenia) gjorgjevici Hadzisce, 1959, is a junior syn- onym of Pseudamnicola depressa Radoman, 1957. Other problems were caused by the fact that Radoman (1973a) established many nominal genera without a description or defin- ition but only by combining them with avail- able or new included nominal species. Some of these nominal genera were available at the 106 BODON, MANGANELLI & GIUSTI first introduction (Bracenica, Daphniola, Is- lamia and Strugia), others were only made available subsequently (Dolapia, Prespoli- torea and Zaumia), and others have never been made available (Naumia, Ohridostu- ranya and Rotondia). Key to acronyms in figures: AG, albumen gland; BC, bursa copulatrix; C, ctenidium; CG, capsule gland; CS, cuticularized struc- ture (stylet); DSR, distal (first or RS1) seminal receptacle; FP, fecal pellets; GPD, gonoperi- cardial duct; HG, hypobranchial gland; |, in- testine; LO, loop of the oviduct; MP, muscular pleat; OC, opercular crest; OE, oesophagus; OP, opercular peg; OS, osphradium; OT, op- ercular thickening; P, penis; PD, penial duct; PG, prostate gland; PGL, penial glandular lobe; PL, penial lobe; PSR, proximal (Second or RS2) seminal receptacle; PW, posterior wall of pallial cavity; R, rectum; RC, mass of refringent cells; S, stomach; SG, seminal groove; VD, vas deferens; VE, vas efferens (seminal vesicle). A SURVEY OF THE EUROPEAN VALVATIFORM HYDROBIID GENERA Arganiella Giusti & Pezzoli, 1980 Arganiella Giusti & Pezzoli, 1980: 45. Type Species: Arganiella pescei Giusti & Pez- zoli, 1980, by monotypy. Arganiella pescei Giusti & Pezzoli, 1980 Arganiella pescei Giusti & Pezzoli, 1980: 45-46, fig. 19A-C. Type Locality: “reticoli di falda delle Marche, dell’Abruzzo e del Lazio (in questa re- gione limitatamente alla provincia di Rieti)”, Italy. Restricted by Giusti & Pez- Zoli (1981: 213) to: “Pozzo 163, lungo la SS 150, 32 m sim, 42°30'13”N, 01°27'55’E (versante orientale dell’Ap- pennino centrale, in provincia dell’Aquila, Abruzzo [Well no. 163, along state road no. 150, Km 9.8, altitude 32 m, 42°30'13"N, 01°27’55”E, western side of central Apennines, province of L'Aquila, Abruzzo, Italy]” (actually, the restricted type locality is not in L'Aquila, but in Ter- amo). Type Material: holotype (shell) is in the Giusti collection, Siena, Italy; three paratypes (SMF 254290, shells) are at the Sen- ckenberg-Museum, Frankfurt am Main, Germany, and others (shells and pre- served specimens) in the Giusti and Pez- zoli collections, Milan, Italy (Giusti & Pez- zoli, 1981). Material Examined —Well no. P/163 along the state road 150, km 9.8, Teramo, Abruzzo, ltaly, 33T VH 12, R. Argano 4 G. L. Pesce leg. 3.5.1975 (2 males, 1 female, some shells) (Pesce 4 Silverii, 1976). —Well no. P/161 near S. Petronilla, Roseto, Teramo, Abruzzo, Italy, 33T VH 12, R. Argano & G. L. Pesce leg. 3.5.1975 (3 males, 3 females, many specimens) (Pesce & Silverii, 1976). —Well no. P/42, state road 17, Contrada Buc- cella, L'Aquila, Abruzzo, Italy, 33T UG 69, С. |. Pesce leg. 16.12.1972 (1 male, 1 specimen) (Argano et al., 1975). —Well no. P/37, state road 17, turn-off to Sassa, before Raio Stream, L'Aquila, Abruzzo, Italy, 33T UG 59, G. L. Pesce leg. 9. 1976 (3 males, 5 females, many specimens) (Argano et al., 1975). —S. Susanna springs, Rivodutri, Rieti, Latium, Italy, 33T UH 2307, M. Bodon leg. 12.4.1993 (5 shells), M. Bodon 4 С. Manganelli leg. 29.6.1995 (2 males, 1 fe- male, 7 shells). —Well no. R/23, Via Salaria, km 88.8, Rieti, Latium, Italy, 33T UG 29, G. L. Pesce leg. 13.6.1973 (1 female) (Pesce & Fusac- chia, 1975). —Well no. R/28, Madonnella, Via Cicolana, Km 5.6, Rieti, Latium, Italy, 33T UG 29, G. L. Pesce leg. 13.6.1973 (1 male, 1 specimen) (Pesce & Fusacchia, 1975). For other localities where only shells have been found see Giusti & Pezzoli (1981). Description Shell very small, valvatiform to planispiral, thin, pale whitish, glassy, transparent when fresh; surface of protoconch malleated; spire from rather raised to almost flat, consisting of 2.75-3 rather rapidly growing convex whorls; last whorl large, dilated, more or less de- scending near aperture; umbilicus wide; aper- ture prosocline, roundish, sometimes pyri- form; peristome complete, slightly thickened, slightly reflected only at lower and columellar margin (Figs. 1, 10; Giusti & Pezzoli, 1980: 8 >= FIGS. 1-9. Shell, operculum and anatomical details of Arganiella pescei Giusti & Pezzoli, 1980, from well no. P/37, state road 17, cross-roads for Sassa, before the Raio stream, L'Aquila, Abruzzo, Italy, G. L. Pesce leg. 9.1976 (Figs. 1-7) and shell and operculum of Daphniola exigua (Schmidt, 1856) from the spring Daphne in the Tembe valley, Thessalia, Greece, 7.1980, ex W. J. M. Maassen collection (Figs. 8-9). Figs. 1, 8: shell; Figs. 2, 9: outer face (left), profile (centre in Fig. 2; right in Fig. 9) and inner face (right in Fig. 2) of operculum; Fig. 3: body of a male with pallial cavity open to show head and penis; Figs. 4, 5: penis of three males, dorsal side (left in Figs. 4, 5) and ventral side (right in Fig. 4); Fig. 6: renal and pallial oviduct, intes- tine and pallial organs of a female; Fig. 7: male genitalia (penis and testis excluded), intestine and pallial or- gans. Scale bar = 1 mm. 108 BODON, MANGANELLI & GIUSTI 45, fig. 19A; 1981: 208-209, figs. 3, 7). Di- mensions: height = 0.85-1.05 mm; diameter = 1.65-2.00 mm. Operculum thin, yellowish white or yellow- ish orange, paucispiral, slightly thickened and often with a circular thickening at centre of inner face (Fig. 2; Giusti & Pezzoli, 1980: 45; 1981: 208, 213, fig. 2E). Body unpigmented; eye spots absent (Fig. 3; Giusti & Pezzoli, 1980: 45, fig. 19B; 1981: 208, 213, figs. 1F, 2A). Male genitalia with prostate gland bulging slightly into pallial cavity; penis from short to elongated, flat, with sides corrugated, slightly dilated then tapering near apex, usually end- ing in very pointed tip; penis without lobes, but sometimes with a crest or a small swelling on ventral side near base; penial duct zig-zag- ging through right portion of penis, opening at penis tip (Figs. 4, 5, 7; Giusti & Pezzoli, 1980: 45, fig. 19; 1981: 208-209, fig. 1F-O). Female genitalia with distal seminal recep- tacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle rather de- veloped, arising from oviduct close to point of origin of duct of bursa copulatrix; bursa copu- latrix large, sac-like or kidney-shaped, with rather long duct entering on anterior side; seminal groove running along ventral side of capsule gland (Fig. 6; Giusti & Pezzoli, 1980: 45, fig. 19C; 1981: 208-209, fig. 1A-C). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its cutting edge V-like, with long robust central denticle and 5-6 smaller denticles on both sides in de- creasing order of size; two basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth apically en- larged, their cutting edge with 9-11 denticles, central of which longer, larger; first marginal teeth with long lateral wing and elongated apex, its cutting edge with long row of 20-22 small denticles; second marginal teeth with long, slender lateral wing and roundish, spoon-like apex with cutting edge carrying rather long row of 18-20 very small denticles (Giusti 8 Pezzoli, 1980: 45; 1981: 208-229, 212, figs. 5A-D). Stomach without posterior caecum; intes- tine with well-developed, U- or S-like bend on pallial wall (Figs. 6, 7; Giusti & Pezzoli, 1980: fig. 19C; 1981: 213, figs. 1A, B, 2A-D, FP). Osphradium variable in size, oval or kidney- shaped; ctenidium consisting of 9-18 lamel- lae (Figs. 6, 7; Giusti & Pezzoli, 1980: 45, fig. 19C; 1981: 208, 213, figs. 1A-B, 2A, F). Nervous system unknown. Taxonomy The genus Arganiella is characterized by: shell very small, valvatiform to planispiral; op- erculum without peg; penis without lobes; fe- male genitalia with distal seminal receptacle and large, sac-like or kidney-shaped bursa copulatrix with anterior duct; central tooth with two pairs of basal cusps. It includes only the type species, which is endemic to the central Apennine, Italy. Another species, the French Valvata exilis Paladilhe, 1867, was erro- neously assigned to this genus by Bouchet (1990) (see “Description of a new valvatiform genus from France”). Bracenica Radoman, 1973a Bracenica Radoman, 1973a: 7, 20. Type Species: Bracenica spiridoni Radoman, 1973a, by monotypy. Bracenica spiridoni Radoman, 1973a Bracenica spiridoni Radoman, 1973a: 7, 20. Type Locality: “Spirov izvor, Podmeret, near Braceni, not far from Virpazar, Crna Gora”, Montenegro. Type Material: lectotype (BEO 116, shell) at the Prirodnjacki Muzej u Beograd to- gether with a paralectotype (BEO 117, shell) (Jovanovic, 1991). Description Shell very small, valvatiform-planispiral; surface of protoconch unknown; spire de- pressed, consisting of 2.75-3.25 rather rapidly growing convex whorls, first whorls slightly raised, last whorl large; umbilicus very wide, deep; aperture roundish to oval; peris- tome complete, thin, slightly reflected only at its lower and columellar margin (Radoman, 1973a: 20; Radoman, 1983: 65, pl. 4, fig. 58; Jovanovic, 1991: pl. 4, fig. 26). Dimensions: height = 0.92-1.18 mm; diameter = 1.64-2.02 mm (Radoman, 1983: table 4). Operculum thin, whitish with yellow central part and peg on inner face (Radoman, 1983: 66). Body unpigmented; (Radoman, 1983: 66). Male genitalia with penis elongated and pointed, with one, evident, knob-like lobe on left side about half way along penis (Rado- man, 1973a: 20; 1983: 66, fig. 29). Female genitalia with two seminal recepta- eye spots absent REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 109 cles and a bursa copulatrix arising from distal renal oviduct; proximal seminal receptacle very small; distal seminal receptacle very large, club-shaped; bursa copulatrix large, pyriform, with long slender duct that enters bursa on anterior side (Radoman, 1973a: 6, 20; 1983: 40, 66, fig. 29). Radula with central tooth with one pair of basal cusps; other details unknown (Rado- man, 1973a: 6; 1983:40). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6; 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1983: 66). Taxonomy Radoman (1973a: 7, 20) established Bracenica without a description or definition but only giving a combined description of it and a single included new nominal species. This makes Bracenica available (ICZN, 1999: Art. 13.4). Bracenica is a little known genus character- ized by: shell very small, valvatiform-planispi- ral; operculum with peg; penis with one simple lobe; female genitalia with two seminal recep- tacles, distal larger than proximal, and large, pyriform bursa copulatrix with anterior duct. It contains only the type species from Montene- gro. Dabriana Radoman, 1974 Dabriana Radoman, 1974: 81. Type Species: Dabriana bosniaca Radoman, 1974, by original designation. Dabriana bosniaca Radoman, 1974 Dabriana bosniaca Radoman, 1974: 81-84, figs. 1-3A-D. Type Locality: “Dabarska [DabarSka] pecina, neben dem Ursprung des Flússchen Dabar, etwa 6 Km súdlich der Stadt San- ski most, Bosnien”. According to Rado- man (1983: 168), the type locality is “Dabarska [DabarSka] pecina (cave), by the source of the Dabar river, about 6 km south of the Sanski Most town”. Type Material: the holotype and six paratypes (SMF 232168) are at the Senckenberg- Museum, Frankfurt am Main, Germany (Jovanovié, 1991). Description Shell very small, valvatiform, whitish, trans- parent when fresh; microsculpture of proto- conch unknown, surface of teleoconch with fine radial striations; spire moderately raised, consisting of 3.25-3.5 rapidly growing convex whorls; last whorl dilated, slightly descending near aperture; umbilicus moderately wide; aperture large, prosocline, roundish; peris- tome complete, slightly reflected only at col- umellar margin (Radoman, 1974: 81, fig. 1; 1983: 167-168, pl. 11, figs. 201, 202; Jo- vanovic, 1991: pl. 10, fig. 6). Dimensions: height = 1.85-2.18 mm; diameter = 2.06-2.49 mm (Radoman, 1983: 208, table 9). Operculum probably without outgrowth, though not specified (Radoman, 1974: fig. 3A; 1983: fig. 102A). Body unpigmented; eye spots absent; cau- dal tentacle at posterior apex of foot (Rado- man, 1974: 81, figs. 1, 3A; 1983: 168, fig. 102A, pl. 11, fig. 202). Male genitalia with penis conical elongated, without lobes; penial apex slender, pointed (Radoman, 1974: 82, fig. 3A, B; 1983: 168, fig. 102A, B). Female genitalia with a seminal receptacle (probably distal) and a bursa copulatrix; sem- inal receptacle very large, ovoid, protruding posteriorly, with evident duct inserted halfway between end of oviduct loop and bursa copu- latrix duct; bursa copulatrix very small, ellipti- cal, with short duct entering bursa on anterior side; capsule gland divided in two parts, the anterior of which narrow; seminal groove run- ning all along ventral side of capsule gland (Radoman, 1974: 81-82, fig. 3C, D; 1983: 167-168, fig. 102 C, D). Radula with central tooth trapezoidal with long lateral wings and basal tongue, its apical margin with robust central denticle and about 6 smaller denticles on both sides in decreas- ing order of size; no basal cusp at point where lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with about 6 large denticles, central of which larger; first marginal teeth rake-shaped, with a long lateral wing and elongated cutting edge with a long row of about 14 small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lateral wing and roundish, spoon-like apex, cutting edge of which carrying rather long row of small denticles (Radoman, 1974: 81, fig. 2; 1983: fig. 101). Stomach without posterior caecum; intes- 110 BODON, MANGANELLI & GIUSTI FIGS. 10-16. Microsculpture of protoconchs. Fig. 10: Arganiella pescei Giusti 8 Pezzoli, 1980, from well no. P/37, state road 17, cross-roads for Sassa, before the Raio stream, L'Aquila, Abruzzo, Italy, G. L. Pesce leg. 9.1976; Fig. 11: Daphniola exigua (Schmidt, 1856) from the spring Daphne in the Tembe valley, Thessalia, Greece, 7.1980, ex W. J. M. Maassen collection; Fig. 12: Hauffenia erythropomatia (Hauffen, 1856) from “Babja Luknja” cave, S. 35, Goricane, Medvode, Slovenia, M. Bodon leg. 16.6.1985; Fig. 13: Fissuria boui Boeters, 1981, from the spring La Foux, Draguignan, Var, France, M. Bodon leg. 7.1.1990; Fig. 14: Haaziella ephippiostoma Киббег, 1932, from the springs MoGilnik, Vrhnika, Slovenia, M. Bodon leg. 17.6.1985; Figs. 15, 16: Hauffenia tellinii (Pollonera, 1898) from the spring between Spagnut and Biacis, upper Natisone val- ley, Pulfero, Udine, Friuli-Venetia Julia, Italy, M. Bodon leg. 9.6.1985. Scale bar = 100 um. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS ah tine unknown (Radoman, 1974: 82; 1983: 166). Osphradium elliptical; ctenidium absent (Radoman, 1974: 81; 1983: 168). Nervous system with very long pleuro- supraoesophageal and short pleuro-suboe- sophageal connectives (Radoman, 1974: 81-82; 1983: 166). Taxonomy Dabriana is a little-known genus character- ized by: shell very small, valvatiform; opercu- lum without peg; foot with caudal tentacle; penis without lobes; female genitalia with large, probably distal, seminal receptacle and very small, elliptical bursa copulatrix with an- terior duct; central tooth of radula without basal cusps. It contains only the type species from Bosnia-Herzegovina. Radoman (1983) dubiously places this genus in the Lithoglyphulidae, but Bole & Velkovrh (1986) and Kabat & Hershler (1993) assigned it to the Hydrobiidae. It shares some characters (central tooth of the radula without basal cusps; foot with caudal tentacle) with some marine rissooids. Dalmatella Velkovrh, 1970 Dalmatella Velkovrh, 1970: 97, 103. Type Species: Dalmatella sketi Velkovrh, 1970, by original designation. Dalmatella sketi Velkovrh, 1970 Dalmatella sketi Velkovrh, 1970: 97-98, 103, fig. 1A-D. Type Locality: “izvir pri odtoku Krke izpod elektrarne pod Skradinskim Bukom (Sibenik, Dalmacija)”, Croatia. It corre- sponds to the “spring under power sta- tions near Skradinski Buk on the river Krka” (Bole & Velkovrh, 1986: 190). Type Material: holotype (9075/1) in the Velkovrh collection, Ljubljana, Slovenia, together with three paratypes (9075/2-4) (Velkovrh, 1970). Description Shell very small, valvatiform, keeled, con- vex above, rather flat below, transparent when fresh; microsculpture of protoconch un- known; spire rather raised, consisting of 3.66 rather rapidly growing whorls; last whorl large, trapezoidal in outline, with marked, rather sharp peripheral keel at base, slightly dilated, descending near aperture; umbilicus wide; aperture prosocline, oval to roundish, with sort of beak at keel; peristome complete, sin- uous, not thickened, slightly reflected only at lower and columellar margin (Velkovrh, 1970: 97-98, 103, fig. 1A-D; Bole € Velkovrh, 1986, fig. 15). Dimensions: height = approximately 1.6 mm; diameter = approximately 2.2 mm (Velkovrh, 1970: 98). Operculum and anatomy unknown. Taxonomy A relatively unknown genus, including only the type species and another undescribed en- tity (Bole & Velkovrh, 1986), both from Croa- tia. Dalmatella is listed as a distinct taxon, but due to lack of anatomical data, its validity is doubtful. Daphniola Radoman, 1973a Daphniola Radoman, 1973a: 8. Type Species: Daphniola graeca Radoman, 1973a, by monotypy. Daphniola graeca is a junior synonym of Valvata exigua Schmidt, 1856, according to Schütt (1980), and a junior synonym of Valvata (Cincinna) hellenica Westerlund, 1898, according to Reischútz 8 Sattmann (1993). Daphniola exigua (Schmidt, 1856) Valvata exigua Schmidt, 1856: 160. Type Locality: “Griechenland”. Following the designation of the neotype by Schütt (1980), the type locality becomes “Thes- salien: mehrere kleine Quellen im Tem- petal in der Náhe der Bahnstation Agia Paraskeui”. Type Material: neotype (SMF 262352, shell) in the Senckenberg-Museum, Frankfurt am Main, Germany (Schútt, 1980). Valvata (Cincinna) hellenica Westerlund, 1898: 179. Type Locality: “Griechenland. Vyteria in Arka- dien”. Type Material: lectotype (4667a, shell) in the Naturhistoriska Museet Góteborg, Góte- borg, Sweden, together with two paralec- totypes (4667b, shells) (Reischútz & Sattmann, 1993). Daphniola graeca Radoman, 1973a: 8, 22. FIGS. 17-34. Shell, operculum and anatomical details of Hauffenia erythropomatia (Hauffen, 1856) from the spring below “Babja Luknja” cave, Goricane, Medvode, Slovenia, M. Bodon leg. 16.6.1985 (Figs. 17-22) and of Fissuria boui Boeters, 1981, from Fontaine de Vaucluse, Vaucluse, France, M. Bodon leg. 1.12.1984 (Figs. 23-26, 28, 33), Source du Vivier, N.D. de Vaucluse, Auribeau-sur-Siagne, Alpes Maritimes, France, M. Bodon leg. 1.1.1991 (Figs. 27, 32), the spring La Foux, Draguignan, Var, France, M. Bodon leg. 7.1.1990 (Figs. 29, 30), the spring La Fouan, Chateauneuf Grasse, Alpes Maritimes, France, M. Bodon, E. Bo & M. Sosso leg. 11.2.1994 (Fig. 31), the spring at the Gorges Mal Infernet, Agay, Var, France, M. Bodon leg. 31.12.1990 (Fig. 34). Figs. 17, 23: shell; Figs. 18, 26: outer face (left in Figs. 18, 25) and profile (right in Fig. 18) of operculum; Figs. 19, 24: body of a male with pallial cavity open to show head and penis; Figs. 20, 25: prostate gland, stomach (excluded in Fig. 25), intestine and pallial organs of a male; Figs. 21, 27: gonadal (excluded in Fig. 21), renal and pallial oviduct, intestine and pallial organs of a female; Fig. 22: penis; Figs. 28, 29: renal and pallial oviduct in three females; Figs. 30-34: penis of four males, dorsal side (left) and ven- tral side (right). Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 113 Type Locality: “spring Daphne, about 30 km north of Larissa, Greece”. Type Material: lectotype (BEO 177, shell) in the Prirodnjacki Muzej u Beograd, to- gether with a paralectotype (BEO 178, shell) (Jovanovic, 1991). Material Examined —Daphne spring in Tembe valley, Thessalia, Greece, 7. 1980, ex W. J. M. Maassen collec- tion (5 shells with dried soft parts). Description Shell very small, valvatiform-globose coni- cal; surface of protoconch malleated; spire well raised, consisting of 3-3.5 rather rapidly growing convex whorls; umbilicus small; aper- ture roundish to oval; peristome complete, its external margin thin, its columellar margin thickened, reflected (Figs. 8, 11; Schutt, 1962: 164, fig. 5, as Horatia (Horatia) exigua; Schutt, 1980: 139-140, pl. 10a, fig. 41, as Ho- ratia (Daphniola) exigua; Radoman, 1973a: 22, as Daphniola graeca; Radoman, 1983: 84-85, pl. 5, fig. 87, as Daphniola graeca; Jo- vanovic, 1991: pl. 6, fig. 44, as Daphniola graeca; Reischutz & Sattmann, 1993, pl. 8a, as Daphniola hellenica). Dimensions: height = 1.22-1.52 mm; diameter = 1.30-1.40 mm (according to Schutt, 1962: 164, as Horatia (Horatia) exigua); height = 1.1-1.3 mm; diam- eter = 1.0-1.2 mm (according to Radoman, 1983: 203, table 5, as Daphniola graeca). Operculum thin, yellowish brown, paucispi- ral, slightly thickened at centre, without out- growth on inner face (Fig. 9; Schutt, 1962: 163, as Horatia (Horatia) exigua; Radoman, 1973a: 22, as Daphniola graeca; Schútt, 1980: 140, as Horatia (Daphniola) exigua). Body pigmented; eye spots present (Schutt, 1962: 163, as Horatia (Horatia) ex- igua). Male genitalia with penis elongated and pointed, with slender lobe on left side at about half penis length (Radoman, 1973a: 22, as Daphniola graeca; Schutt, 1980: 140, as Hor- atia (Daphniola) exigua; Radoman, 1983: 83-84, fig. 45, as Daphniola graeca). Female genitalia with two seminal recepta- cles and a bursa copulatrix (according to Radoman, 1973a, 1983; only one seminal re- ceptacle and bursa copulatrix, according to Schütt, 1980) arising from distal renal oviduct; proximal seminal receptacle markedly larger than distal; bursa copulatrix very large, oval, with long slender duct entering bursa on ante- rior side; seminal groove running along ven- tral side of capsule gland (Radoman, 1973a: 6, 22, as Daphniola graeca; Schutt, 1980: 140, as Horatia (Daphniola) exigua; Rado- man, 1983: 40, 83, fig. 45, as Daphniola graeca). Radula with central tooth with one pair of basal cusps; other details unknown (Schütt, 1980: 140, as Horatia (Daphniola) exigua; Radoman, 1983: 40, as Daphniola graeca). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6, as Daph- niola graeca; Radoman, 1983: 40, as Daph- niola graeca). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and somewhat shorter pleuro-sub- oesophageal connectives (Radoman, 1983: 83, fig. 45, as Daphniola graeca). Taxonomy Radoman (1973a: 8, 22) established Daph- niola without a description or definition but gave a combined description of it and a single included new nominal species which makes this nominal genus available (ICZN, 1999: Art. 13.4). Thus, Daphniola Schütt, 1980 (type species: Valvata exigua Schmidt, 1856), is a junior homonym and a junior synonym of Daphniola Radoman, 1973a. Daphniola is here considered a distinct genus. However, some of its anatomical de- tails are unknown and its relationships to other Balkan genera (Horatia Bourguignat, 1887, in particular), require further study. Daphniola is characterized by: shell very small, valvatiform; operculum without peg; penis with one simple lobe; female genitalia with two seminal receptacles, proximal larger than distal, and very large, oval bursa copula- trix with anterior duct. According to Schutt (1980), Daphniola graeca Radoman, 1973a (p. 22) (type locality: “spring Daphne, about 30 km north of Larissa, Greece”) is a junior synonym of Valvata ex- igua Schmidt, 1856. Reischútz & Sattmann (1993) claimed that the identity of the last nominal taxon was uncertain and proposed to use, for this species, Valvata (Cincinna) hel- lenica Westerlund, 1898. However, since Schútt (1980) designated a neotype for Schmidt’s species, its identity is without prob- lems. Another Greek entity, Horatia (Daphniola) 114 BODON, MANGANELLI & GIUSTI exigua pangaea Reischútz, 1984, has been assigned to Daphniola. As the anatomy of this entity has never been studied, its inclusion in this genus requires confirmation. Erythropomatiana Radoman, 1978 Erythropomatiana Radoman, 1978: 35. Type Species: Valvata erythropomatia Hauf- fen, 1856, by original designation. Erythropomatiana erythropomatia (Hauffen, 1856) Valvata erythropomatia Hauffen, 1856: 465. Type Locality: “Górzaher Grotte (Gorizane)” [=“Babja Luknja” cave], Slovenia. Type Material: type material is in the “Ver- sammlung der Musealmitglieder des Laibacher Museums,” Ljubljana, Slove- nia (Hauffen, 1856). Material Examined —“Babja Luknja” cave, $. 35, Goricane, Med- vode, Slovenia, 33T VM 51, M. Bodon leg. 16.6.1985 (2 females). —Spring below “Babja Luknja” cave, fed by waters from the same cave, Goricane, Medvode, Slovenia, 33T VM 51, M. Bodon leg. 16.6.1985 (1 male, 3 females, 10 shells). —“Marijno Brezno” or “Velika Gipsovka” cave, S. 6, Skofja Loka, Slovenia, 33T VM 4613, F. Stoch leg. 11.1.1998 (1 juv. specimen). Description Shell very small, valvatiform, thin, pale whitish, waxen, transparent when fresh; sur- face of protoconch malleated; spire rather flat, consisting of 2.75-3.25 rather rapidly growing convex whorls; last whorl dilated, slightly de- scending near aperture; umbilicus wide; aper- ture prosocline, roundish; peristome com- plete, thin, slightly reflected only at columellar margin (Figs. 12, 17; Radoman, 1978: 35-36, pl. 5, figs. 18-19; 1983: 123, pl. 9, fig. 146, tab. 7; Bole 4 Velkovrh, 1986: fig. 18). Dimen- sions: height = 0.73-1.13 mm; diameter = 1.17-1.55 mm. Operculum thin, yellowish, paucispiral, slightly thickened but without outgrowth on inner face (Figs. 18, 35; Radoman, 1978: 35; 1983: 123; Bole, 1993: 8, fig. 2B). Body unpigmented; eye spots usually ab- sent (only one out of six specimens examined had eye spots) (Fig. 19; Bole, 1963: 121). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather short, flat, with apex blunt and 2-3 (or one, accord- ing to Bole, 1993: fig. 1C), small, rather evi- dent knob-like lobes on left side near apex; penial duct zig-zagging through central por- tion of penis to open at penis tip; large oval mass of refringent cells inside penis apex to right of penial duct; terminal portion of penial duct (immediately before opening) with very small stylet (Figs. 20, 22; Radoman, 1978: 35, fig. 6; 1983: 123, fig. 68; Bole, 1993: 8, fig. 1C). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle, very small and with very short duct arising from oviduct level with end of loop; bursa copulatrix reduced, small but slightly longer than semi- nal receptacle, not or slightly dilated at apex, arising very close to point at which oviduct en- ters albumen gland portion of pallial oviduct; seminal groove running along entire ventral side of capsule gland (Fig. 21; Radoman, 1978: 35, fig. 6; 1983: 123, fig. 68; Bole, 1993: 8, fig 1B). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long, robust central denti- cle and 5-6 smaller denticles on both sides in decreasing order of size; 1-2 basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 11-13 denticles, central of which longer larger; first marginal teeth rake-shaped with long lateral wing and elongated cutting edge with long row of 23-25 small denticles anteriorly; sec- ond marginal teeth scraper-shaped, with long slender lateral wing and roundish, spoon-like cutting edge, its cutting edge carrying rather long row of 15-19 very small denticles (Figs. 44-45; Radoman, 1978: 35; Bole, 1993: 8, fig. 2A). Stomach without posterior caecum; intes- tine with well developed, tightly coiled, S-like bend on pallial wall (Figs. 20, 21; Bole, 1963: 122, fig. 2B; Radoman, 1978: 35; 1983: 40). Osphradium variable in size, oval or elon- gated, kidney-shaped; ctenidium consisting of 6-11 lamellae (Figs. 20, 21; Bole, 1963: fig. 3C; 1993: 8, fig. 1A). REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 115 Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1978: 35; 1983: 123). Taxonomy Erythropomatiana is here considered a ju- nior synonym of Hauffenia Pollonera, 1898 (see “The status and relationships of Hauffe- nia’). Radoman (1978) introduced this genus for Valvata erythropomatia Hauffen, 1856. Al- though the anatomy of this species was nearly the same as that of the Hauffenia species, Erythropomatiana was regarded as distinct from the latter by Radoman (1983), Bole & Velkrovh (1986), and Bole (1993). Radoman (1978) included in Erythropoma- tiana another species from a different site, E. verdica Radoman, 1978, distinguished only by shell characters. This species is here rec- ognized as a junior synonym of Hauffenia subpiscinalis (KuSkéer, 1932) (see below). Fissuria Boeters, 1981 Fissuria Boeters, 1981: 57-58. Type Species: Fissuria boui Boeters, 1981, by original designation. Fissuria boui Boeters, 1981 Fissuria boui Boeters, 1981: 58-59, figs. 5-9, pl. 6, figs. 5-7. Type Locality: “Frankreich, Dép. Vaucluse bzw. Bouches du Rhône, Durance- Grundwasser”. Type Material: the holotype (SMF 253580) is at the Senckenberg-Museum, Frankfurt am Main, Germany; paratypes are at the Senckenberg-Museum, Frankfurt am Main, Germany (SMF 253581), at the Nationaal Natuurhistorisch Museum, Lei- den, The Nederlanas, and in the Boeters collection (124, 418, 761), Múnchen, Germany (Boeters, 1981). Material Examined —Fontaine de Vaucluse, Vaucluse, France, M. Bodon leg. 1.12.1984, 22.6.1989 (3 males, 6 females, 8 shells). —Debris of Durance River near Orgon, Bouches du Rhóne, France, M. Bodon leg. 1.12.1984 (28 shells). —La Foux spring, Draguignan (Var, France), 32T KP 92, M. Bodon leg. 7.1.1990 (4 males, 4 females, many shells). —Spring at the Gorges Mal Infernet, Agay, Var, France, 32T LP 21, M. Bodon leg. 31.12.1990 (2 males, 11 females). —Source du Vivier, N.D. de Vaucluse, Au- ribeau-sur-Siagne, Alpes Maritimes, France, 32T LP 3231, M. Bodon leg. 1.1.1991 (5 males, 5 females, many shells). —La Fouan spring, Chateneuf Grasse, Alpes Maritimes, France, 32T LP 3737, M. Bodon, E. Bo & M. Sosso leg. 11.2.1994 (6 males, 9 females, many shells). — Source de la Foux, Mouans-Sartoux, Alpes Maritimes, France, M. Bodon leg. 2.1.1999 (2 females, many shells). — Alluvial springs in the bed of the Var River, on the right bank, 100-200 m upstream of wells, la Tuiliere, St-Laurent du Var, Alpes Maritimes, France, M. Bodon leg. 2.1.1999 (3 males, 1 shell). Description Shell very small, valvatiform, thin, whitish, glassy, transparent when fresh; surface of protoconch malleated; spire from well raised to almost flat, consisting of 2.75-3.5 rather rapidly growing convex whorls; last whorl rather large, slightly dilated descending near aperture; umbilicus of variable width; aperture prosocline, roundish to ovoid; peristome com- plete, rather thin, slightly reflected only at lower and columellar margin (Figs. 13, 23; Boeters, 1981: 58, pl. 6, figs. 5-7). Dimen- sions: height = 1.19-1.61 mm; diameter = 0.93-1.77 mm. Operculum thin, yellowish, paucispiral, not thickened and without outgrowth on inner face (Fig. 26). Body unpigmented (a few traces of pigment in wall of visceral sac); eye spots absent (Fig. 24). Male genitalia with prostate gland slightly bulging into pallial cavity; penis rather short, flat, with apex pointed and 3-4 (rarely 2) more or less evident raised lobes containing mass of glandular tissue; lobes of variable size and position: usually two lobes on left side and one on ventral side at about 2/3 of penis length; sometimes also one lobe on dorsal side near base of penis); penial duct zig-zag- ging through right portion of penis to open at penis tip (Figs. 25, 30-34; Boeters, 1981: 57-58, figs. 5-8). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal and distal seminal re- 116 BODON, MANGANELLI 8 GIUSTI ceptacles more or less equal in size; bursa copulatrix variable, from very small (Boeters, 1981) to rather large, oval, with proportionally more or less elongated, slender duct entering bursa on anterior side; seminal groove run- ning along ventral side of capsule gland (Figs. 27-29; Boeters, 1981: 57-58, fig. 9). Radula with central tooth trapezoidal with long lateral wings and basal tongue; anterior margin with 9-11 denticles, central of which longer and larger; two basal cusps, outer of which very small, at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their ante- rior margin with 10-11 denticles, central of which longer and larger; first marginal teeth rake-shapea, with long lateral wing and elon- gated cutting edge with long row of 22-26 small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lat- eral wing and roundish spoon-like apex, its cutting edge carrying rather long row of 20-24 very small denticles (Figs. 46, 47). Stomach without posterior caecum; intes- tine with rather developed, tightly coiled, S- like bend on pallial wall (Figs. 25, 27; Boeters, 1981, fig. 8). Osphradium variable in size, more or less elongated, oval or kidney-shaped; ctenidium consisting of 6-13 lamellae (Figs. 25, 27). Nervous system unknown. Taxonomy The genus Fissuria is characterized by: shell very small, valvatiform; operculum with- out peg; penis with 2-4 glandular lobes; fe- male genitalia with two seminal receptacles equal in size and bursa copulatrix oval and of variable size, with anterior duct; central tooth with two pairs of basal cusps. Beyond the type species, Fissuria includes an undescribed species from Liguria, Italy (Pezzoli, 1988a; Bodon et al., 1995b). Other two additional species are tentatively as- signed to this genus: “Fissuria” planospira Bodon, Cianfanelli & Talenti, 1997, from Tus- cany, Italy, by Bodon et al. (1997) and Horatia (Hauffenia) raehlei Schütt, 1980, from Cephalonia |., Greece, in this paper (see below). Gocea Hadzisce, 1956 Gocea Hadzisce, 1956: 496-499. Type Species: Gocea ohridana Hadzisce, 1956, by original designation. vıvv Gocea ohridana Hadzisce, 1956 Gocea ohridana Hadzisée, 1956: 496-499, figs. 1-4. Type Locality: “Ohridsee. Die Schnecke lebt in einem bis jetzt fast ununtersuchten Biotop des Sees, nämlich an den steini- gen Zonen der litoralen Region, und zwar nicht an den oberflächigen Steinen, son- dern tiefer unten an solchen... .”, Mace- donia. According to Radoman (1983: 81) the type locality is: “Lake Ohrid on the stones by Veli Dab, spreading the east lake bank”. Type Material: Hadzisce (1956) did not give any information about the type material. Material Examined — Trepcja south of Ohrid, Lake Ohrid, Mace- donia, 34T DL 83, ex W. J. M. Maassen collection (2 shells with dried soft parts). Description Shell very small, valvatiform, ram-horn-like, partly despiralized; surface of protoconch pit- ted; spire from rather raised to almost flat, consisting of 2.75-3.25 rather rapidly growing convex whorls; last whorl large, despiralized, descending; umbilicus (in spiralized part of shell) rather large; aperture prosocline, irreg- ularly pyriform due to sinuous contour (upper margin extended forward, angled at upper vertex; external margin convex, very slightly angled at periphery; lower margin slightly ex- tended forward; columellar margin concave); peristome complete, thin, slightly reflected only at columellar margin (Fig. 53; Hadzisce, 1956: 496-497, 499, figs. 1a-b, 2a; HadZiSce et al., 1976: 2-3, figs. 1-4; Maassen, 1980: pl. 17, figs. 29-30; Radoman, 1983: 81, pl. 5, figs. 79, 80; Bole 8 Velkovrh, 1986: fig. 20). Dimensions: height = 0.55-0.85 mm; diame- ter = 1.20-1.60 (Hadzisce, 1956: 499); height = 0.53-0.85 mm; diameter = 0.60-1.55 mm (Radoman, 1983: 203, table 5). Operculum reddish, multispiral and pecu- liarly spiralized on outer face to resemble screw; foot tissue penetrating hole at centre of inner face (Fig. 54; Hadzisce, 1956: 497-498, fig. 26; HadziSce, 1959: 87; Hadzisce et al., 1976, fig. 1; Radoman, 1983: 81, fig. 42A). Body slightly pigmented; eye spots present (Hadzisce, 1956, fig. 4). Male genitalia with penis elongated and rather pointed, with slightly raised but evident REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS dali FIGS. 35-43. Operculum and opercular structures of Hauffenia erythropomatia (Hauffen, 1856) from the spring below “Babja Luknja” cave, Goricane, Medvode, Slovenia, M. Bodon leg. 16.6.1985 (Fig. 35); Hauf- fenia tellinii (Pollonera, 1898) from the spring Perilo, near Robic, Nadiza valley, Slovenia, М. Bodon leg. 9.6.1995 (Figs. 36, 37); Kerkia kusceri (Bole, 1961) from the spring of the Krka River, Krka, Slovenia, M. Bodon leg. 16.6.1985 (Figs. 38, 39); Pseudohoratia ochridana (Polinski, 1929) from Lake Ohrid, Macedonia, ex P. Radoman collection (Fig. 40), Hauffenia subpiscinalis (Kuscer, 1932) from the “ZelSe Jame” caves, $. 576, near Rakek, Slovenia, 3.3.1966, ex F. Velkovrh collection (Fig. 41) and Hauffenia wagneri (Kuscer, 1928) from the spring of the “Vranja pec” cave, BoStanj, Sevnica, KrSo, Slovenia, M. Bodon leg. 14.6.1985 (Figs. 42, 43). Figs. 35, 36, 38, 40-42: inner face of operculum; Figs. 37, 43: detail of opercular peg; Fig. 39: detail of opercular crest. Scale bar = 100 um. 118 ВОРОМ, MANGANELLI & GIUSTI hidis. FIGS. 44-52. Radula of Hauffenia erythropomatia (Hauffen, 1856) from “Babja Luknja” cave, S. 35, Goricane, Medvode, Slovenia, M. Bodon leg. 16.6.1985 (Figs. 44, 45), Fissuria boui Boeters, 1981, from Fontaine de Vaucluse, Vaucluse, France, M. Bodon leg. 1.12.1984 (Figs. 46, 47),Hauffenia tellinii (Pollonera, 1898) from the spring between Spagnut and Biacis, upper Natisone valley, Pulfero, Udine, Friuli-Venetia Julia, Italy, M. Bodon leg. 9.6.1985 (Figs. 48, 49) and Hauffenia subpiscinalis ( Kuscer, 1932) from the spring Obrh, Gorenje Jezero, Cerknica, Slovenia, M. Bodon leg. 19.6.1985 (Figs. 50-52). Figs. 44, 46, 48, 50: cen- tral part of radula; Figs. 45, 47: outer marginal teeth; Figs. 49, 52: lateral, inner and outer marginal teeth; Fig. 51: three central teeth. Scale bar = 5 um. lobe on left side near apex (as deduced from Hadzisce, 1956: 499, fig. 4a, and from Rado- man’s, 1983; fig. 42D; but according to Rado- man’s, 1983, description: “penis long, cylindri- cal, smooth, without any outgrowth”) (Hadzisce, 1956: 499, fig. 4a; Radoman, 1983: 81, fig. 42D). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal and distal seminal re- ceptacles about same size; bursa copulatrix large, kidney-shaped, with long slender duct entering bursa on anterior side; seminal groove running all along ventral side of cap- sule gland (Radoman, 1983: 81, fig. 42B, C). Radula with central tooth trapezoidal with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 11119 WZ 39 54 | AS CN Os 55 96 A FIGS. 53-59. Shell and operculum of Gocea ohridana Hadziëée, 1956, from Trepcja south of Ohrid, Lake Ohrid, Macedonia, ex W. J. M. Maassen collection (Figs. 53, 54) and shell, operculum and anatomical de- tails of Hadziella ephippiostoma Киббег, 1932, from the Moëilnik springs, Vrhnika, Slovenia, М. Bodon leg. 17.6.1985 (Fig. 55) and the spring at Lozice, Deskle, Soéa [Isonzo] valley, Slovenia, M. Bodon leg. 10.7.1996 (Figs. 56-59). Figs, 53, 55-56: shell; Figs. 54, 57: outer face (left), profile (centre in Fig. 54; right in Fig. 57) and inner face (right in Fig. 54) of operculum; Fig. 58: penis; Fig. 59: male genitalia (penis and testis ex- cluded), intestine and pallial organs. Scale bar = 1 mm. 120 BODON, MANGANELLI & GIUSTI and 4-5 smaller denticles on both sides in de- creasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 10-11 denticles, central of which longer and larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of about 20 denticles anteriorly; second marginal teeth scraper-shaped, with long slender lateral wing and roundish spoon- like apex, its cutting edge carrying rather long row of small denticles (HadZisce, 1956: 498, fig. 3; Hadzisce et al., 1976: 13, figs. 18, 19). Stomach without posterior caecum; intes- tine unknown (Radoman, 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and somewhat shorter pleuro- suboesophageal connectives (Radoman, 1983: 81). Taxonomy Gocea is here considered a distinct genus. However some of its anatomical details are unknown, and its relationships to other Balkan genera require further study. Gocea is char- acterized: by shell very small, valvatiform, ram-horn-like, partly despiralized; operculum without peg but spiralized on outer face to re- semble screw; penis with one simple lobe; fe- male genitalia with two seminal receptacles equal in size and large, kidney-shaped bursa copulatrix with anterior duct; central tooth with one pair of basal cusps. It includes only the type species, which is endemic to Lake Ohrid. Hadziella Kuscer, 1932 Haaziella Kuscer, 1932: 54. Type Species: Hadziella ephippiostoma Kuscer, 1932, by monotypy. Haaziella ephippiostoma Kuscer, 1932 Haaziella ephippiostoma Kuscer, 1932: 54- 55, pl. 3, figs. 4a, b. Type Locality: “Quelle bei Podgora”, Ljubljan- ica basin, Slovenia. Type Material: holotype (2004a) in the Kuscer collection, Institute of Biology, University of Ljubljana, Ljubljana, Slovenia (Kuscer, 1932). Material Examined —Mo6cilnik springs, Vrhnika. Plentiful karstic springs which feed the Ljubljanica River, Slovenia, 33T VL 49, M. Bodon leg. 17.6.1985 (32 shells). — Springs near the right bank of Verd Stream, upstream of Verd village, Vrhnika. Plenti- ful karstic springs, Slovenia, 33T VL 48, M. Bodon leg. 17.6.1985 (9 shells). —Obrh spring, Gorenje Jezero, Cerknica. Plentiful karstic spring in the Cerknisko Lake basin (the waters flow underground to feed the Rak River), Slovenia, 33T VL 56, M. Bodon leg. 19.6.1985 (6 shells). —Cemun spring, Gorenje Jezero, Cerknica, Slovenia, 33T VL 56, M. Bodon leg. 19.6.1985 (3 shells). —Spring near LoZice, along road to Kanal, Soéa valley, Slovenia, 33T UM 9201, M. Bodon leg. 10.7.1996 (1 male, many shells). Description Shell very small, planispiral, concave on both sides, whitish, glassy, transparent when fresh; microsculpture of protoconch consist- ing of spiral rows of minute, more or less elon- gated knobs; spire flat, consisting of 3-3.25 rather rapidly growing whorls; external wall of last whorl flat obliquely with sort of obtuse keel at base, large, dilated, descending slightly near aperture; aperture prosocline, oval-triangular (slightly angled between col- umellar and external, external and lower and lower and columellar margin); peristome com- plete, thickened, reflected (Figs. 14, 55, 56; Kuscer, 1932: 54-55, pl. 3, figs. 4a, b; Bole, 1963: 124, Fig. 4A, B; Maassen, 1975: pl. 27, figs. 9, 10; Bole & Velkovrh, 1986: fig. 21). Di- mensions: height = 0.51-0.56 mm; diameter = 1.43-1.75 mm. Operculum thin, multispiral, concave, with very reduced outgrowth at centre of inner face (Fig. 57; Bole, 1993: 13, 15, fig. 3D). Body unpigmented (a few traces of pigment in wall of visceral sac); eye spots absent. Male genitalia with entire prostate gland bulging into pallial cavity; penis slightly elon- gated, conical, with sides corrugated, tapering near apex, ending in pointed tip; penial duct zig-zagging through right portion of penis to open at penis tip (Figs. 58, 59; Bole, 1993: 13, 15, fig: SC): Female genitalia with only one (?) sac-like structure (bursa copulatrix) arising from distal REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 121 66 FIGS. 60-66. Shell, operculum and anatomical details of Hadziella anti Schutt, 1960, from the “Bus de l'Ors” cave, no. 64 FR, Cornappo valley, Udine, Friuli-Venetia Julia, Italy, 33T UM 6923, M. Bodon 8 F. Stoch leg. 23.7.96. Fig. 60: shell; Fig. 61: body of a female with pallial cavity open to show head; Fig. 62: outer face (left), profile (centre) and inner face (right) of operculum; Fig. 63: dorsal side of penis; Fig. 64: male genitalia (penis and testis excluded), intestine and pallial organs; Figs. 65, 66: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar = 1 mm. 122 BODON, MANGANELLI 8 GIUSTI renal oviduct; bursa copulatrix large, shoe- shaped, with rather long, wide duct entering bursa on posteroventral side; seminal recep- tacle absent (?): according to Bole (1993), its function carried out by portion of renal oviduct level with end of loop (from where proximal seminal receptacle usually arises); seminal groove running along ventral side of capsule gland (Bole, 1993: 13, 15, fig. 3C). Radula with central tooth trapezoidal with long lateral wings and basal tongue, its apical margin with long robust central denticle and about 4 smaller denticles on both sides in de- creasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with many small denticles, central of which longer and larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of small denticles anteriorly; second marginal teeth scraper-shaped, with long and slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of very small denticles (Bole, 1993: 13, 15, fig: SE). Stomach without posterior caecum; intes- tine with a twisted bend near the style sac and well developed, tightly coiled, Z-like bend on pallial wall (Fig. 59; Bole, 1993: 13, 15, fig. 3B). Osphradium oval; hypobranchial gland well developed; ctenidium absent (Fig. 59; Bole, 1993: 13, 15, fig. 3A). Nervous system unknown. Taxonomy The genus Hadziella is characterized by: shell very small, planispiral, concave on both sides; operculum with very reduced peg; penis without lobes; female genitalia with dis- tal seminal receptacle and large, shoe- shaped bursa copulatrix with posteroventral duct; central tooth with one pair of basal cusps. The description of the female anatomy of Haaziella is entirely based on Bole's (1993) study of the type species. Dissection of fe- males of H. anti (Figs. 60-66) confirmed some of the characters ascertained by Bole (1993) (bursa copulatrix large, shoe-shaped, duct entering bursa on posteroventral side), but also revealed others that were not de- scribed (pallial oviduct and bursa copulatrix entirely inside the pallial cavity). Some char- acters seem to distinguish H. anti (oviduct loop absent; seminal receptacle arising from renal oviduct close to point from where duct of bursa copulatrix arises) from H. ephippios- toma. Because H. anti and H. ephippiostoma are closely related (they share many shell and anatomical characters), the seminal recepta- cle and oviduct loop are presumably similar as well. The type species should therefore be reinvestigated. Apart from the type species, Hadziella in- cludes six other species from northeastern Italy and the northwestern Balkans: H. anti Schutt, 1960; H. deminuta Bole, 1961; H. krkae Bole, 1992; H. rudnicae Bole, 1992; H. sketi Bole, 1961; and H. thermalis Bole, 1992 (Bole & Velkovrh, 1986; Bole, 1992); another undescribed entity is cited by Bole & Velkovrh (1986). Hauffenia Pollonera, 1898 Hauffenia Pollonera, 1898: 3, as a subgenus of Horatia. Type Species: Horatia (Hauffenia) tellinii Pol- lonera, 1898, by subsequent designation (Walker, 1918). Hauffenia tellinii (Pollonera, 1898) Horatia (Hauffenia) tellinii Pollonera, 1898: 3-4, fig. 2. Type Locality: “Lalta valle del Natisone nel Friuli”, Italy. Type Material: Pollonera (1898) did not give any information about the type material. The Pollonera collection is at the Museo Regionale di Scienze Naturali di Torino, but the syntypes of this species have not been traced (E. Gavetti, pers. com., 27.11.1997). Horatia (Hauffenia) valvataeformis Pollonera, 1898: 3-4, fig. 3. Type Locality: “l'alta valle del Natisone nel Friuli”, Italy. Type Material: Pollonera (1898) did not give any information about the type material. The Pollonera collection is at the Museo Regionale di Scienze Naturali di Torino, but the syntypes of this nominal taxon have not been traced (E. Gavetti, pers. com., 27.11.1997). Hauffenia michleri Kuscer, 1932: 56-57, pl. 5, fig. 3. Type Locality: “Ljubljanica quellen Mocilnik”, Slovenia. Type Material: holotype (2005a) in the KuScer collection, “Zoologischen Institut der Uni- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 123 FIGS. 67-80. Shell, operculum and anatomical details of Hauffenia tellinii (Pollonera, 1898) from debris of Natisone River, Paderno, Premariacco, Udine, Friuli-Venetia Julia, Italy, M. M. Giovannelli leg. 5. 1988 (Figs. 67, 70), from the spring Perilo, near Robic, Nadiza valley, Slovenia, M. Bodon leg. 9.6.1995 (Figs. 68, 80), from the spring between Spagnut and Biacis, upper Natisone valley, Pulfero, Udine, Friuli-Venetia Julia, Italy, M. Bodon leg. 23.9.1993, 9.6.1985 (Figs. 69, 72-74, 77, 79), from the spring at Tarpezzo, Alberone valley, S. Pietro al Natisone, Udine, Friuli-Venetia Julia, Italy, M. Bodon leg. 1.4.1991 (Fig. 71) and from the springs of Timavo River, S. Giovanni al Timavo, Duino-Aurisina, Trieste, Friuli-Venetia Julia, Italy, M. Bodon leg. 31.3.1991 (Figs. 75, 76, 78). Fig. 67: shell; Figs. 68-71: outer face (Fig. 68 left), profile (Fig. 68 centre; Figs. 69-71) and inner face (Fig. 68 right) of operculum; Fig. 72: body of a male with pallial cavity open to show head and penis; Figs. 73-75: dorsal side of penis of eight males; Figs. 76, 77: prostate gland, stomach, in- testine and pallial organs of two males; Figs. 78, 79: renal and pallial oviduct, intestine and pallial organs of two females; Fig. 80: renal and pallial oviduct of a female. Scale bar = 1 mm (67-72, 74-80); = 0.5 mm (73). 124 BODON, MANGANELLI & GIUSTI FIGS. 81-86. Shell, operculum and anatomical details of Hauffenia michleri KuScer, 1932, (junior synonym of H. tellinii) from the Mocilnik springs, Vrhnika, Slovenia, M. Bodon leg. 17.6.1985 (Fig. 81) and the spring on right bank of the Verd brook, near the Verd spring, Vrhnika, Slovenia, M. Bodon leg. 17.6.1985 (Figs. 82-86). Fig. 81: shell; Fig. 82: outer face (left), profile (centre) and inner face (right) of operculum; Fig. 83: body of a male with pallial cavity open to show head and penis; Fig. 84: dorsal side (first picture on left and last two on right) and ventral side (second picture) of penis of three males; Fig. 85: prostate gland, stomach, intestine and pallial organs of a male; Fig. 86: renal and pallial oviduct, intestine and pallial organs of a fe- male. Scale bar = 1 mm. versitat Ljubljana”, Slovenia (KusScer, 1932). Material Examined —Perilo spring, Robic, upper Nadiza [Nati- sone] valley, Slovenia, 33T UM 8522, M. Bodon leg. 9.6.1995 (1 female, many shells). —Alluvial springs on left bank of the Nadiza River, 250 m upstream from the gorge, Hurja, Potoki, Slovenia, 33T UM 8323, M. Bodon, S. Cianfanelli & G. Manganelli leg. 25.7.1996 (1 male, 1 female, 4 shells). —Alluvial springs on the right bank of the Nadiza River, at the km 37 milestone on the road to Kobarid, Slovenia, 33T UM 8422, M. Bodon 4 1. Misic leg. 15.7.1996 (1 male). —Plentiful spring 1.1 km from the Italy-Slove- nia border, welling out below the road in the upper Natisone valley (Pulfero, Udine, Friuli-Venetia Julia, Italy), 33T UM REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 125 8318, M. Bodon leg. 1.4.1991 (1 female, many shells). —Spring between Spagnut and Biacis, upper Natisone valley, Pulfero, Udine, Friuli- Venetia Julia, Italy, 33T UM 8211, M. Bodon leg. 23.9.1993, 9.6.1985, 1.4.1991 (11 males, 3 females, many shells) (Pezzoli, 1988a). — Cave of $. Giovanni d’Antro no. 43 Fr, Pulfero, Udine, Friuli-Venetia Julia, Italy, 33T UM 8212, M. Bodon leg. 22.9.1983 (many shells), F. Gasparo & F. Stoch leg. 19.7.1992 (2 females, 3 shells). —Mustig spring, Vedronza, Torre valley, Lu- severa, Udine, Friuli-Venetia Julia, Italy, 33T UM 6524, M. Bodon, M. M. Giovan- nelli & F. Stoch leg. 22.7.1989 (1 female, 10 shells). —Spring at Tarpezzo, Alberone valley, welling below the road and flowing through the village, S. Pietro al Natisone, Udine, Friuli-Venetia Julia, Italy, 33T UM 8411, М. Bodon leg. 8.6.1985 (4 shells), 1.4.1991 (4 males, 1 female, many shells) (Pezzoli, 1988a, cited, in part, as Islamia sp.). —Plentiful spring 1 km downstream from Bod- igoi, Ludrio valley, welling below the road, Prepotto, Udine, Friuli-Venetia Julia, Italy, 33T UM 8402, M. Bodon leg. 1.4.1991 (1 female, many shells). —Spring 700 m upstream of Podresca, Ludrio valley, Prepotto, Udine, Friuli-Venetia Julia, Italy, 33T UM 8905, M. Bodon leg. 7.7.1993 (3 males, 1 female, many shells). — Spring near Podclanz at the turn-off for Salamant, Ludrio valley, Prepotto, Udine, Friuli-Venetia Julia, Italy, 33T UM 9107, M. Bodon leg. 7.7.1993 (1 male, 1 fe- male, many shells). — Alluvial springs on the right bank of the Torre River, 100-200 m upstream from of the confluence of Isonzo, Papariano, Fiumi- cello, Udine, Friuli-Venetia Julia, Italy, 33T UL 7774, M. Bodon leg. 27.7.1996 (1 male, 5 females, 5 shells). —Alluvial springs on the right bank of the Isonzo River, under the railway viaduct, Papariano, Fiumicello, Udine, Friuli- Venetia Julia, Italy, 33T UL 7774, M. Bodon leg. 27.7.1996 (2 males and 1 fe- male). — Springs of the Timavo River, S. Giovanni al Timavo, Duino-Aurisina, Trieste, Friuli- Venetia Julia, Italy, 33T UL 9071, M. Bodon leg. 31.3.1991 (6 males, 2 fe- males, many shells) (Stammer, 1932; Pezzoli, 1988a). —Alluvial spring near Molin del Cucco, Rivoli di Osoppo, Osoppo, Udine, Friuli-Venetia Julia, Italy, 33T UM 4921, S. Cianfanelli & M. Calcagno leg. 29.12.1998 (1 male). —“Grotta de la Foos” cave, no. 229 Fr, Cam- pone, Tramonti di Sotto, Pordenone, Friuli-Venetia Julia, Italy, 33T UM 3124, M. Bodon & F. Stoch leg. 7.7.1991, 15.7.1993 (1 male, 1 female, 2 shells) (Pezzoli, 1988b, 1989). —“Grotta | Landri” cave, no. 1254 V, Colli di Conegliano, San Pietro di Feletto, Tre- viso, Venetia, Italy, 33T TL 8585, F. Stoch leg. 15.7.1999 (1 juv. specimen). —Spring on the right bank of the Idrijca Stream, near Ukovnik, Spodnja Idrija, Slovenia, 33T VM 20, F. Gasparo e F. Stoch leg. 5.6.1993 (1 male, 1 female, 1 shells). —Mocilnik springs, the main Ljublianica source near Vrhnika (type locality of Hauffenia michleri), Slovenia, 33T VL 49, M. Bodon leg. 17.6.1985 (1 female, many shells). —Springs on the right bank of Verd brook, Vrhnika, Slovenia, 33T VL 48, M. Bodon leg 17.6.1985 (3 males, 1 females, many shells). Other shells with operculum were examined from the following localities: — Spring “La Santissima” of the Livenza River, Polcenigo, Pordenone, Friuli-Venetia Julia, Italy, 33T UL 0499, M. Bodon leg. 28.3.1991 (1 shell) (Pezzoli, 1988a). —Debris of Barcis lake, Barcis, Pordenone, Friuli-Venetia Julia, Italy, 33T UM 1318, M. Bodon leg. 8.7.1991 (1 shell). —Spring on the right bank of a stream affluent of the Pontaiba Stream, Colle di Pinzano, Pinzano, Pordenone, Friuli-Venetia Julia, Italy, 33T UM 4118, M. Bodon 4 F. Stoch leg. 15.7.1993 (1 shell). —Spring along the road upstream Casiacco, Arzino valley, Vito d'Asio, Pordenone, Friuli-Venetia Julia, Italy, 33T UM 4221, M. Bodon & F. Stoch leg. 15.7.1993 (1 shell). —Debris of the Tagliamento River, Ponte della Delizia, Valvasone-Codroipo, Por- denone-Udine, Friuli-Venetia Julia, Italy, 33T UL 3792, M. Bodon leg. 27.4.1986 (1 shell) (Pezzoli, 1988a). —Peschiera spring, Vedronza, Torre valley, 126 BODON, MANGANELLI & GIUSTI Lusevera, Udine, Friuli-Venetia Julia, Italy, 33T UM 6524, M. Bodon leg. 24.9.1983 (1 shell) (Pezzoli, 1988a). —Springs on left bank of Vedronza River, Casera Morandin, Lusevera, Udine, Friuli-Venetia Julia, Italy, 33T UM 6425, M. Bodon leg. 24.9.1983 (1 shell) (Pez- zoli, 1988a). —Debris of the Natisone River, Paderno, Pre- mariacco, Udine, Friuli-Venetia Julia, Italy, 33T UM 7600, M. M. Giovannelli, leg. 5.1988 (1 shell). —Spring between Robic and Suzid, Slovenia, 33T UM 8721, M. Bodon leg. 9.6.1985 (1 shell). —Springs downstream from Liessa, Cosizza valley, Grimacco, Udine, Friuli-Venetia Julia, Italy, 33T UM 9012, M. Bodon leg. 1.4.1991 (1 shell). —Springs on right bank of the Grivo Stream, Canal di Grivo, Faedis, Udine, Friuli- Venetia Julia, Italy, 33T UM 7314, M. Bodon leg. 8.7.1983 (1 shell). —Debris of the Isonzo River, Peteano, Sagrado d’lsonzo, Gorizia, Friuli-Venetia Julia, Italy, 33T UL 8783, M. Bodon leg. 20.6.1985 (1 shell) (Bodon & Giovannelli, 1994; Pezzoli, 1988a). —Debris of the Isonzo River, Papariano, Fiu- micello, Udine, Friuli-Venetia Julia, Italy, 33T UL 7774, M. Bodon leg. 21.9.1983, 20.6.1985 (5 shells) (Bodon & Giovan- nelli, 1994; Pezzoli, 1988a). —Spring on right bank of the Idrijca Stream, upstream from Podroteja, Idrija, Slove- nia, 33T VL 29, M. Bodon leg. 13.7.1996 (5 shells). Shell material from the type locality was col- lected in the following place: —Debris of the Natisone River, upstream Lin- der, Pulfero, Udine, Friuli-Venetia Julia, Italy, 33T UM 8216, M. Bodon leg. 23.9.1983 (8 shells) (Pezzoli, 1988a). For other localities where only shells were col- lected see Pezzoli (1988a, 1996). Description Shell very small, valvatiform to planispiral, thin, pale whitish, waxen, transparent when fresh; surface of protoconch malleated; spire from rather raised to almost flat, consisting of 2.5-3.5 rapidly growing convex whorls; last whorl large, dilated, more or less descending, sometimes slightly detached near aperture; umbilicus wide, about 1/6-1/3 of shell di- ameter; aperture more or less prosocline, roundish to pyriform; peristome complete, thin and not or slightly reflected only at columellar margin (Figs. 15, 16, 67, 81, 87; Pollonera, 1898: 3, figs. 2, 3, as Horatia valvataeformis; Kuscer, 1932: 56-57, pl. 5, fig. 3, as Hauffe- nia michleri; Bole, 1967b: 112, fig. 1A, as Hauffenia michleri; Bole, 1970: 4-6, fig. 1; Maassen, 1975: pl. 27, figs. 4-8, as Hauffenia michleri; Pezzoli et al., 1975: pls. 1, 2; Rado- man, 1978: 34-35, pl. 4, figs. 13-15, as Hauf- fenia michleri; Bole, 1979: 36, figs. 1-4a, b, as Hauffenia michleri; Giusti & Pezzoli, 1980: 45, figs. 18B-D, 26L; Giusti & Pezzoli, 1982: pl. 1, fig. 4; Radoman, 1983: 122, table 7, pl. 9, figs. 143, 144, as Hauffenia michleri; Bole & Velkovrh, 1986: fig. 22; Pezzoli, 1989, pl. 5, fig. 11; Bodon & Giovannelli, 1994: fig. 4M, N). Dimensions: height = 0.45-1.20 mm; diame- ter = 0.98-2.30 mm. Operculum yellowish, paucispiral, thick and with well developed spiralized peg at centre of inner face; peg variable in size in different populations, but always dilated at apex (Figs. 36, 37, 68-71, 82; Pollonera, 1898: 3, fig. 2, fig. 3, as Horatia valvataeformis; Bole, 1967a: 88, fig. 4(5), as Hauffenia michleri; Bole, 1967b: 113, fig. 1B3, as Hauffenia michleri; Bole, 1970: 91, fig. 2A5; Bole, 1993: 6, fig. 2B; Radoman, 1978: 33-34, as Hauffenia mich- leri; Radoman, 1983: 121-122, as Hauffenia michleri). Body unpigmented (a few traces of pigment sometimes on visceral sac); eye spots absent (Figs. 72, 83; Bole, 1967a: 87-88, fig. 4(2), as Hauffenia michleri; Bole, 1967b: 112-113, as Hauffenia michleri). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather short, flat, with apex blunt and no or 1-2 slightly ev- ident lateral lobes on left side near apex, pe- nial duct zig-zagging through central portion of penis to open at penis tip; globular mass of refringent cells inside penis apex to right of penial duct; terminal portion of penial duct (immediately before opening) with very small stylet (Figs. 73-77, 84-85, 95-99; Bole, 1967a: 87, fig. 4(4), as Hauffenia michleri; Bole, 1967b: 112, fig. 1B2, as Hauffenia mich- leri; Bole, 1970: 91, fig. 2A3; Giusti & Pezzoli, 1980: 45, fig. 18E; Radoman, 1983: 120, as Hauffenia michleri; Bole, 1993: 6, fig. 1C, as Hauffenia michleri). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; the smaller structure con- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 127 tains spermatozoa oriented with their heads anchored to cells of the wall and is thus a seminal receptacle, and the larger contains few non-oriented and partially digested sper- matozoa and is thus a bursa copulatrix; sem- inal receptacle very small and sessile or with very short duct arising from oviduct level with end of loop; bursa copulatrix very small but markedly longer than seminal receptacle, not dilated at apex, arising very close to point at which oviduct enters albumen gland portion of pallial oviduct; seminal groove running along ventral side of capsule gland (Figs. 78-80, 86, 101; Bole, 1967a: 87-88, fig. 4(3), as Hauffenia michleri; Bole, 1970: 91, fig. 2A2; Giusti & Pezzoli, 1980: 43, fig. 18F; Rado- man, 1983: 40, 120, as Hauffenia michleri; Bole, 1993: 6, fig. 1B; Bole, 1993: 6, as Hauf- fenia michleri). Radula with central tooth trapezoidal with long lateral wings and basal tongue, its apical margin V-like, with long robust central denti- cle and 5-6 smaller denticles on both sides in decreasing order of size; 1-2 basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth rake- like, apically enlarged, their anterior margin with 10-13 denticles, central of which longer and larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 16-26 small denticles anteriorly; second marginal teeth scraper- shaped, with long slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 15-18 very small denticles (Figs. 48-49; Bole, 1967a: 87, fig. 4(6), as Hauffenia michleri; Bole, 1967b: 112, fig. 1B4, as Hauffenia michleri; Bole, 1970: 91, fig. 2A4; Bole, 1993: 6, fig. 2A). Stomach without posterior caecum; intes- tine with well developed, Z-like or question- mark-like bend on pallial wall (Figs. 76-79, 85-86; Radoman, 1983: 40, as Hauffenia michleri). Osphradium variable in size, oval or elon- gated, kidney-shaped; ctenidium absent or consisting of 1-13 lamellae (Figs. 76-79, 85-86; Bole, 1967a: 87 fig 4(1), as Hauffenia michleri; Bole, 1967b: 112, fig. 1B1, as Hauf- fenia michleri; Bole, 1970: 91, fig. 2A1; Bole, 1993: 6, fig. 1A). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1983: 120, as Hauf- fenia michleri). Taxonomy Hauffenia is the second nominal genus es- tablished for the European valvatiform hydro- biids. It was introduced as a subgenus of Ho- ratia Bourguignat, 1887, and raised to the rank of distinct genus by Киббег (1932, 1933a, b). This was only demonstrated to be correct when Bole (1970) studied the type species anatomically and found its genitalia to be clearly different from those described by Radoman (1966) in the type species of Hora- tia. (For diagnosis, status and relationships of Hauffenia with Erythropomatiana Radoman, 1978, Neohoratia Schutt, 1961, and Vrania Radoman, 1978, see “Status and Relation- ships of Hauffenia”, below.) In the last 30 years, many valvatiform hy- drobiid species from western Europe were in- cluded into Hauffenia (also as Horatia (Hauf- fenia)), often simplistically on the basis of shell characters alone, by Bole (1961), Schútt (1961b, 1980), Binder (1966) and Bernasconi (1975, 1984, 1985). Most of them are in need of revision because only the shells were stud- ied. Some are revised and redescribed in the present paper. This revision enabled us to as- certain that geographical distribution of Hauf- fenia is limited to the northern sector of the former Yugoslavia (Slovenia and Croatia), Austria and the neighbouring areas of north- eastern Italy. Наийета michleri is here recognized as ju- nior synonym of H. tellinii (see taxonomic re- marks to H. tellinii in the section on Hauffenia species). Horatia Bourguignat, 1887 Horatia Bourguignat, 1887: 46, 47-49. Type Species: Horatia klecakiana Bourguig- nat, 1887, by subsequent designation (Westerlund, 1902). Horatia klecakiana Bourguignat, 1887 Horatia klecakiana 1887: 49-50, figs. 18-21. Type Locality: “sorgente pres de Ribaric, dans la vallée de la Cettina”, Croatia. Accord- ing to Radoman (1983: 52) this corre- sponds to “the Vrijovac spring in the source area of the Cetina river”. Type Material: lectotype (shell) in the Bour- guignat collection, Museum d'Historie Naturelle de Geneve, Geneva, Switzer- land (Binder, 1957). Bourguignat, 128 BODON, MANGANELLI & GIUSTI FIGS. 87-94. Microsculpture of protoconchs. Fig. 87: Hauffenia michleri Киббег, 1932, (junior synonym of H. tellinii) from the Mocilnik springs, Vrhnika, Slovenia, M. Bodon leg. 17.6.1985; Fig. 88: Horatia klecakiana Bourguignat, 1887, from Cetina near Vrlika, Croatia, 16.9.1964, ex F. Velkovrh collection; Fig. 89: Kerkia kusceri (Bole, 1961) from the spring of the Krka River, Krka, Slovenia, M. Bodon leg. 16.6.1985; Fig. 90: Hauffenia subpiscinalis (Kuscer, 1932) from the spring Kotla, Rakov Skocjan, Slovenia, M. Bodon leg. 18.6.1985; Fig. 91: Pezzolia radapalladis Bodon & Giusti, 1986, from the spring inside the Rio di Tonnego, Ponte della Vittoria, Rapallo, Genova, Liguria, Italy, M. Bodon leg. 16.12.1979; Fig. 92: Pseudohoratia ochri- dana (Polinski, 1929) from Lake Ohrid, Macedonia, ex P. Radoman collection; Fig. 93: Hauffenia wagneri (Kuscer, 1928) from the spring of the “Vranja Pec” cave, Bostanj, Sevnica, KrSko, Slovenia, M. Bodon leg. 14.6.1985; Fig. 94: Hauffenia subcarinata Bole 8 Velkovrh, 1987, from the spring near LoZice, along the road to Kanal, Soca valley, Slovenia, M. Bodon leg. 10.7.1996. Scale bar = 100 um. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 129 PL Vy ee RC 102 109 = FIGS. 95-103. Penis, stylet and pallial oviduct of Hauffenia tellinii (Pollonera, 1898) from the spring between Spagnut and Biacis, upper Natisone valley, Pulfero, Udine, Friuli-Venetia Julia, Italy, M. Bodon leg. 23.9.1993, 1.4.1991 (Figs. 95, 96, 98, 101), Hauffenia michleri KuSéer, 1932, (junior synonym of H. tellinii) from the spring on right bank of the Verd brook, near the Verd spring, Vrhnika, Slovenia, M. Bodon leg 17.6.1985 (Figs. 97, 99), and Hauffenia subpiscinalis (Киббег, 1932) from the “ZelSke Jame” caves, $. 576, near Rakek, Slovenia, 3.3.1966, ex Е. Velkovrh collection (Fig. 100) and from the Mocilnik springs, Vrhnika, Slovenia, M. Bodon leg. 17.6.1885 (Figs. 102, 103). Figs. 95-97: penis; Figs. 98, 99: apex of penis; Fig. 100: the entire stylet; Figs. 101, 102: section of capsule gland; Fig. 103: detail of seminal groove. Scale bar = 100 um (95-97), 50 um (98, 99, 101, 102), 10 um (100, 103). 101 130 BODON, MANGANELLI & GIUSTI PSR 110 FIGS. 104-110. Operculum, shell and anatomical details of Horatia klecakiana Bourguignat, 1887, from Cetina near Vrlika, Croatia, 16.9.1964, ex F. Velkovrh collection. Fig. 104: outer face (left), and profile (right) of operculum; Fig. 105: shell; Fig. 106: body of a male with pallial cavity open to show head and penis; Fig. 107: stomach; Fig. 108: renal and pallial oviduct, intestine and pallial organs of a female; Fig. 109: prostate gland, intestine and pallial organs of a male; Fig. 110: dorsal side of penis of three males. Scale bar = 1 mm. Material Examined —Cetina near Vrlika, Croatia, 33T Xd, 16.9.1964, ex F. Velkovrh collection (3 males, 2 females, many shells). Description Shell very small, valvatiform to ovoid, rather robust, whitish, opaque; surface of proto- conch malleated; spire more or less raised, consisting of 3.25-3.5 rather rapidly growing convex whorls; last whorl dilated and more or less descending near aperture; umbilicus not very wide; aperture prosocline, roundish to oval; peristome complete, external margin thin, columellar margin thickened, reflected (Figs. 88, 105; Bourguignat, 1887: 47-56, figs. 2, 3, as Horatia letourneuxi, figs. 4, 5, as H. praeclara, figs. 6, 7, as H. albanica, figs. 10, 11, as H. obliqua, figs. 10-13, as H. ver- likana, figs. 14, 15, as H. palustris, figs. 16, 17, as H. fontinalis, figs. 18-23, as H. obtusa; Binder, 1957: 59-62, figs. 1a-r, u-w; Ant, 1962: 74, figs. 1-21, 25-30; Radoman, 1965: 143, Figs. 3, 4; Radoman, 1966: 246, fig. 2; Radoman, 1983: 52, pl. 3, fig. 38). Dimen- sions: height = 1.55-2.14 mm; diameter = REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 131 1.81-2.18 mm (Radoman, 1965: table 2; 1983: 201, table 3). Operculum thin, reddish yellow, paucispiral, slightly thickened at centre, lacking outgrowth on inner face (Fig. 104; Radoman, 1965: fig. 3; Boeters, 1974: 86; Radoman, 1983: 52). Body slightly pigmented; eye spots present (Fig. 106; Boeters, 1974: 86). Male genitalia with prostate gland bulging well into pallial cavity; penis rather elongated and slender, with apex pointed and single or double, rather evident, lobe on left side at about 2/3 of its length; penial duct zig-zagging through lateral portion of first half of penis, be- coming central before opening at penis tip (Figs. 109-110; Boeters, 1974: 86, figs. 1, 2; Radoman, 1983: 52, fig. 20). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal seminal receptacle thin and bent to adhere to oviduct level with end of loop; distal seminal receptacle larger than proximal, wide at apex; bursa copulatrix large, kidney-shaped, with long slender duct that enters bursa on anterodorsal side; semi- nal groove running along ventral side of cap- sule gland (Fig. 108; Radoman, 1966: 249, fig. 8; 1973a: 6; Boeters, 1974: 86; Radoman, 1983: 40, 51, fig. 20). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long, robust central denti- cle and 5 smaller denticles on both sides in decreasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 10-11 denticles, central of which longer and larger; first marginal teeth rake-shapea, with long lat- eral wing and elongated cutting edge with long row of 25-30 small denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of very small denticles (Binder, 1957: 62, fig. 2; Boeters, 1974: 86; Hershler & Longley, 1986: fig. 28C). Stomach without posterior caecum; intes- tine with bend absent or relatively undevel- oped (Figs. 107-109; Radoman, 1973a: 6; Boeters, 1974: 86; Radoman, 1983: 40). Osphradium variable in size, elongated, kidney-shaped; ctenidium consisting of 7-11 lamellae (Figs. 108, 109; Boeters, 1974: 86). Nervous system with long pleuro-supraoe- sophageal and rather long to somewhat short pleuro-suboesophageal connectives (Rado- man, 1966: 249; 1983: 40, 51) Taxonomy The genus Horatia is characterized by: shell very small, valvatiform to ovoid; opercu- lum without peg; penis with one simple lobe; female genitalia with two seminal receptacles, distal larger than proximal, and large, kidney- shaped bursa copulatrix, with anterodorsal duct; central tooth with one pair of basal cusps. Horatia was the first nominal genus estab- lished for the European valvatiform hydro- biids. Before then, and sometimes also after, valvatiform hydrobiids were included in the heterobranch genus Valvata Muller, 1774. Together with the type species, Bourguig- nat (1887) introduced many nominal species of Horatia. Apart from Horatia servaini Bour- guignat, 1887, which is regarded as a junior synonym of /slamia valvataeformis (Móllen- dorff, 1873) (Radoman, 1983), all the others, from Dalmatia and Albania are currently re- garded as junior synonyms of Horatia kle- cakiana Bourguignat, 1887 (Binder, 1957; Radoman, 1983). They are: Horatia albanica Bourguignat, 1887; H. fontinalis Bourguignat, 1887; H. letourneuxi Bourguignat, 1887; H. obliqua Bourguignat, 1887; H. obtusa Bour- guignat, 1887; H. palustris Bourguignat, 1887; H. praeclara Bourguignat, 1887; and H. ver- likana Bourguignat, 1887. Ant (1962) also regarded H. knorri Schütt, 1961, as a possible junior synonym of H. klecakiana, but this is very doubtful. Besides the type species, Horatia includes two other species inhabiting different drain- ages from that where H. klecakiana lives: H. macedonica (Kuscer, 1936) and H. novose- lensis Radoman, 1966, both from Macedonia (Radoman, 1983). Reports of species of Horatia exist from other European and non-European countries (Willmann & Pieper, 1978; Bole & Velkovrh, 1986; Kabat & Hershler, 1993), but all of them are questionable. Islamia Radoman, 1973a Islamia Radoman, 1973a: 10. Type Species: Horatia servaini Bourguignat, 1887, by original designation. Horatia servaini is a junior synonym of Hydrobia 132 BODON, MANGANELLI & GIUSTI 112 PGL MP 113 FIGS. 111-116. Shell, operculum and anatomical details of Islamia valvataeformis (Möllendorff, 1873) from Vrelo Bosne, Sarajevo, Bosnia, G. Vigna & A. Vigna leg. (Figs. 111-115) and shell of Karevia ornata (Rado- man, 1957) from the harbour of Trpezjca, Lake Ohrid, Macedonia, 5.1976, ex W. J. M. Maassen collection (Fig. 116). Fig. 111: body of a male with pallial cavity open to show head and penis; Figs. 112, 116: shell; Fig. 113: outer face of operculum; Fig. 114: intestine and pallial organs of a male; Fig. 115: dorsal side (left) and ventral side (right) of penis. Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 133 valvataeformis Mollendorff, 1873, ac- cording to Radoman (1983). Islamia valvataeformis (Móllendorff, 1873) Hydrobia valvataeformis Móllendorff, 1873: 59. Type Locality: “. . . an Steinen in der Quelle der Bosna, welche am Fuss des lg- mangebirges bei Sarajevo. . .”, Bosnia. According to Radoman (1983: 126) the type locality corresponds to “Vrelo Bosne, near Sarajevo”. Type Material: Möllendorff (1873) did not give any information about the type material. Horatia servaini Bourguignat, 1887: 52. Type Locality: “Sources de la Bosna, pres de Sérajewo, Bosnie”. Type Material: syntypes (2 shells) in the Bour- guignat collection, Museum d’Historie Naturelle de Genève, Geneva, Switzer- land (Binder, 1957). Material Examined —Vrelo Bosne, Sarajevo, Bosnia, 34T BP 85, G. Vigna & A. Vigna leg. (2 males, 3 shells). Description Shell very small, valvatiform, conical-ovoid, thin, pale whitish, waxen, transparent when fresh and clean; microsculpture of protoconch unknown; spire well raised, consisting of 3.5-4.25 rapidly growing convex whorls; last whorl large, dilated, more or less descending near aperture; umbilicus rather small; aper- ture prosocline, roundish to ovoid; peristome complete, thin, slightly thickened at columellar margin and slightly reflected at lower and col- umellar margin (Fig. 112; Bourguignat, 1887: 52, figs. 8, 9, as Horatia servaini; Radoman, 1973b: 227, 229, fig. 1A, as Islamia (Islamia) servaini; Radoman, 1983: 126, pl. 9, fig. 148). Dimensions: height = 1.62-2.00 mm; diame- ter = 1.53-1.81 mm (Radoman, 1983: Tab. 7). Operculum thin, yellowish, paucispiral, without thickening or outgrowth at centre of inner face (Fig. 113; Radoman, 1973b: 227, as Islamia (Islamia) servaini; Radoman, 1983: 124). Body pigmented; eye spots present (Fig. 111; Giusti et al., 1981: fig. 4.3, as /slamia servaini). Male genitalia with penis large, elongated, dorso-ventrally flat, with cylindrical outline, apically bifid due to well-developed penial lobe on left side, slightly larger than tip of penis proper and containing mass of glandu- lar cells; muscular pleat on ventral side of penis about 2/3 of penis length near base of penial lobe; penial duct zig-zagging through right portion of penis to open at tip of penis proper (Fig. 115; Radoman, 1973b: 227, 231, 233, fig. 3, as Islamia (Islamia) servaini; Giusti et al., 1981: fig. 4.3, as /slamia servaini; Radoman, 1983: 124, 126, fig. 69C). Female genitalia with only two seminal re- ceptacles arising rather close to one another from distal renal oviduct half way between end of loop (where proximal seminal recepta- cle arises in most hydrobiids having two sem- inal receptacles) and where oviduct enters al- bumen gland (near where distal seminal receptacle arises); that arising proximally (closer to end of loop) being rather well devel- oped, always larger and longer than other, usually wider at apex than at base and with short but evident stalk; that arising distally (closer to where oviduct enters albumen gland) being very small and usually without evident stalk; bursa copulatrix absent; semi- nal groove running along ventral side of cap- sule gland (Radoman, 1973b: 227, fig. 2, as Islamia (Islamia) servaini; Radoman, 1973a: 6, 10; 1983: 40, 124, fig. 69A, B). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 5-6 smaller denticles on both sides in de- creasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 9-11 den- ticles, central of which longer and larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 25-28 small denticles anteriorly; sec- ond marginal teeth scraper-shaped, with long and slender lateral wing and roundish, spoon- like apex, its cutting edge carrying rather long row of very small denticles (Radoman, 1973b: 227, fig. 2, as Islamia (Islamia) servaini; Giusti et al., 1981: 53, pl. 3, figs. 1-3, as /slamia ser- vaini; Radoman, 1983: 124). Stomach without posterior caecum; intes- tine with relatively undeveloped, U-like bend on pallial wall (Fig. 114; Radoman, 1973a: 6; Giusti et al., 1981: fig. 4.3, as /slamia servaini; Radoman, 1983: 40). Osphradium variable in size, kidney- shaped; ctenidium consisting of 13-15 lamel- 134 BODON, MANGANELLI & GIUSTI lae (Fig. 114; Giusti et al., 1981: 53, fig. 4.3, as Islamia servain)). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1973b: 227, as Is- lamia (Islamia) servaini; Radoman, 1973a: 6; 1983: 124). Taxonomy Radoman (1973a: 10) established /s/amia without a description or definition, but only by combining it with an available species group name and some new species. After 1930, this condition did not make a generic name avail- able (ICZN, 1999: Art. 13.6). However, at the same time he designated a type species for the new nominal genus and introduced the new family group taxon, Islamiinae, of which he gave a diagnosis. Hence, we can consider this as a “combined description of a new fam- ily-group taxon and a new genus,” which con- fers availability to each name (ICZN, 1999: Art. 13.5). The genus /s/amia is characterized by: shell very small, ovoid to planispiral; operculum without peg; penis with one glandular (rarely non-glandular) lobe; female genitalia with two seminal receptacles, proximal larger than dis- tal, and without bursa copulatrix; central tooth with one or two pairs of basal cusps. Adriolitorea Radoman, 1973b (p. 234; type species: /slamia zermanica Radoman, 1973a) and Mienisiella Schútt, 1991 (p. 134; type species: Mienisiella mienisi Schútt, 1991) are junior synonyms of /slamia Rado- man, 1973a. Adriolitorea was introduced by Radoman (1973b) as a subgenus of /slamia and then regarded as synonym of the latter by Radoman (1983). Mienisiella is considered to be a junior synonym of Islamia on the basis of personal unpublished data. Schútt (1991) de- scribed the female genitalia of M. mienisi Schutt, 1991, and M. gaillardoti (Germain, 1911) as having a very small bursa copulatrix and one seminal receptacle. Study of topo- typical specimens of M. mienisi and others of M. gaillardoti from many localities in Israel confirmed what is clearly evident from Schutt’s (1991: fig. 5c): the two species have two seminal receptacles located and shaped exactly as in the /s/amia species. Penis struc- ture in Mienisiella (see Schutt 1991: 134-135, figs. 5b, 6b) is also exactly the same as that of the /slamia species. In fact, Schutt’s second “small, simple excrescence on left side of penis” is the muscular pleat described above on the penis of /. valvataeformis and present on the penis of many /slamia species studied up to now. Study of M. gaillardoti also showed that the penial duct runs in the right side of the penis to open at the tip of the penis proper (right branch of bifid apex of penis) and not, as figured by Schútt (1991), at the tip of the pe- nial lobe (left branch of bifid apex of penis). Boeters (1998) mistakenly regarded /s- lamia as a junior synonym of Neohoratia. In fact, he did not base this conclusion on the concordance between the type species of Neohoratia and Islamia (Valvata (?) subpisci- nalis Kuscer, 1932, and Horatia servaini Bour- guignat, 1887 [a junior synonym of Hydrobia valvataeformis Mollendorff, 1873], respec- tively), but on the fact that an alleged “Neoho- ratia” species from western Europe actually turns out to be an /slamia species: Islamia cf. minuta (Draparnaud, 1805) (see “Descrip- tions of some taxa misidentified as Hauffenia species” below). A number of species from different sites in Europe and Turkey have been assigned to Is- lamia by Giusti 4 Pezzoli (1980), Giusti et al. (1981), Reischútz (1988), Radoman (1973a, b, 1983) and Bodon et al. (1995a, b). Some of them, assigned to /s/amia on the basis of shell characters, are in need of revision. vevv Karevia Hadzisce, 1959 Karevia Hadzisce, 1959: 81-82, as a sub- genus of Ohrigocea Hadzisce, 1959. Types Species: Ohrigocea (Karevia) prlitchevi Hadzisce, 1959, by subsequent designa- tion (Radoman, 1963a) according to ICZN (1999: Art. 69.2.2). Ohrigocea prl- itchevi is a junior synonym of Karevia or- nata (Radoman, 1957), according to Radoman (1963a, b, 1983). Karevia ornata (Radoman, 1957) Pseudamnicola ornata Radoman, 1957: 88-89, figs. 3, 6, 7C, F, 8. Type Locality: “. . . su uz istoënu оба Ohrid- skog jezera, po obalskom kamenju, pocev od izvora Veli Dab pa do pocetka peskovite, juzne obale jezera, prema Sv. Naumu.”, Lake Ohrid, Macedonia. The type locality is “Lake Ohrid, on the bank stones by Veli Dab. Spread from this lo- cality to a great sandy beach near Sveti Naum”, according to Radoman (1983: 81). Type Material: lectotype (BEO 161, shell) at REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 135 the Prirodnjacki Muzej u Beograd, to- gether with a paralectotype (BEO 162, shell) (Jovanovic, 1991). Ohrigocea (Karevia) prlitchevi Hadzisce, 1959: 83-86, figs. 20, 21. Type Locality: “. . . lebt an seichten steinigen Zonen der Litoralregion auf der nord- östlichen, östlichen und sudöstlichen Seite des Sees [Lake Ohrid], Macedo- nia”. Type Material: HadZisce (1959) did not give any information about the type material. Material Examined —Harbour of Trpezjca, Lake Ohrid, Macedo- nia, 34T DL 83, 5.1976, ex W. J. M. Maassen collection (1 shell with dried soft parts). Description Shell very small, valvatiform-planispiral, adorned with two rows of more or less raised nail-like projections, regularly spaced to follow course of a peripheral keel and basal keel; surface of protoconch malleated; spire de- pressed, consisting of 2.75-3.25 rather rapidly growing whorls; last whorl dilated, de- scending slightly near aperture, its external wall with peripheral keel and its lower wall with basal keel; umbilicus very wide; aperture prosocline, irregularly rhomboidal, angled above and at two keels; peristome complete, thin, slightly reflected only at columellar mar- gin (Fig. 116; Radoman, 1957: 88-89, fig. 3; Hadzisce, 1959: 83-86, figs. 20, 21, as Ohrigocea (Karevia) prlitchevi; Hadzisce, 1959: 86-87, fig. 23, as Ohrigocea (Karevia) sandanskii; Maassen, 1980: pl. 17, figs. 35, 36; Radoman, 1963a: 78-79, fig. 7, as Pseudamnicola (Karevia) ornata; Radoman, 1963b: 95-96, fig. 7, as Pseudamnicola (Karevia) ornata; Radoman, 1983: 80-81, pl. 5, fig. 77, as Dolapia ornata; Jovanovic, 1991: pl. 5, fig. 40, as Dolapia ornata). Dimensions: height = 0.72-1.10 mm; diameter = 1.45-1.73 mm (Radoman, 1983: tab. 5). Operculum yellowish red, paucispiral, prob- ably without peg though not specified. Body pigmented; eye spots present. Male genitalia with penis large, flat with apex rather pointed and lobe on left side about 2/3 of penis length (Radoman, 1957: 89, fig. 7F; HadZiSce, 1959: 86, fig. 20, as Ohrigocea (Karevia) prlitchevi; Hadzisce, 1959: 87, fig. 24, as Ohrigocea (Karevia) san- danskii; Radoman, 1963a: fig. 15, as Pseud- amnicola (Karevia) ornata; Radoman, 1963b: fig. 15, as Pseudamnicola (Karevia) ornata; Radoman, 1983: 80, fig. 41, as Dolapia or- nata). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal seminal receptacle much larger than distal, which is rudimentary; bursa copulatrix large, arched, kidney- or crescent-shaped, with long, slender duct en- tering bursa on anterior side; seminal groove running along ventral side of capsule gland (Radoman, 1957: 89, fig. 7C; Radoman, 1963a: 78, fig. 15, as Pseudamnicola (Kare- via) ornata; Radoman, 1963b: 95, fig. 15, as Pseudamnicola (Karevia) ornata; Radoman, 1973a: 6, as Dolapia ornata; Radoman, 1983: 40, 80, fig. 41, as Dolapia ornata). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 4-5 smaller denticles on both sides in decreasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 9-13 denticles, central of which longer and larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 14-20 denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of very small denticles (Rado- man, 1957: 90, fig. 6; Hadzisce, 1959: 85-86, fig. 22, as Ohrigocea (Karevia) prlitchevi; Hadzisce, 1959: 86-87, fig. 24, as Ohrigocea (Karevia) sandanskil). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6, as Dolapia ornata; Radoman, 1983: 40, as Dolapia or- nata). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1957: 91, fig. 8; Radoman, 1973a: 6, as Dolapia ornata; Rado- man, 1983: 80, as Dolapia ornata). Taxonomy Karevia is here considered a distinct genus, but its relationships to Ohridohauffenia HadZisce, 1959, and Ohrigocea HadZisce, 1959, need to be clarified. Karevia is charac- terized by: shell very small, valvatiform- 136 BODON, MANGANELLI & GIUSTI planispiral, bicarinate and adorned with two rows of nail-like projections; operculum with- out peg; penis with one simple lobe; female genitalia with two seminal receptacles, proxi- mal larger than distal, which is rudimentary, and large, crescent-shaped bursa copulatrix with anterior duct; central tooth with one pair of basal cusps. lt includes only the type species, which is endemic to Lake Ohrid. Dolapia Radoman, 1983 (p. 80, type species: Pseudamnicola ornata, Radoman, 1957), is a junior synonym of Karevia Ha- dzisce, 1959. In fact, apparently overlooking his previous designation of Ohrigocea (Kare- via) prlitchevi Hadzisce, 1959 (as P. ornata, with O. prlitchevi as a junior synonym), as type species of Karevia, Radoman (1973a) designated another type species: Ohrigocea (Karevia) miladinovorum Hadzisée, 1959. At the same time, he introduced a new genus, Dolapia, for P. ornata. However, he (Rado- man, 1973a: 8) established this nominal genus without a description or definition but only by combining it with an available species group name. After 1930, this condition does not make a generic name available (ICZN, 1999: Art. 13.6). Ohrigocea (Karevia) sandanskii Hadzisce, 1959 (pp. 86-87, figs. 23, 24; type locality: “. . . lebt auf der östlichen und sudóstlichen Seite des Sees, in den seichten Teilen seines steinigen Litorals [Lake Ohrid]”, Macedonia figs. 117-122 is another junior synonym of Karevia ornata (Radoman, 1963a, b, 1983). Kerkia Radoman, 1978 Kerkia Radoman, 1978: 29. Type Species: Hauffenia kusceri Bole, 1961, by original designation. Kerkia kusceri (Bole, 1961) Hauffenia kusceri Bole, 1961: 62, 67, fig. 3A. Type Locality: “Krska jama”, Slovenia. Ac- cording to Radoman (1983: 112), the type locality is “Jama Krke (cave), south- east of Ljubljana”. Type Material: Bole (1961) did not give any in- formation about the type material. Material Examined —Spring of the Krka River, Krka, Slovenia, 33T VL 88, M. Bodon leg. 16.6.1985 (1 male, 1 female, many shells). —“KrSka Jama” cave, 5. 74, Krka, Slovenia, 33T VL 88, F. Gasparo & F. Stoch leg. 26.9.1992 (13 specimens, 2 shells). Description Shell very small, valvatiform, thin, whitish; surface of protoconch malleated; spire de- pressed to slightly raised, consisting of 2.75-3.25 rather rapidly growing convex whorls; last whorl dilated, more or less de- scending near aperture; umbilicus wide; aper- ture prosocline, roundish to oval; peristome complete, thin, slightly reflected at columellar margin. (Figs. 89, 117; Bole, 1961: 62, 67, fig. 3A; Radoman, 1978: 29-30, pl. 4, figs. 3, 4; 1983: 110, 112, pl. 8, fig. 128). Dimensions: height = 0.97-1.44 mm; diameter = 1.60-2.34 mm (Radoman, 1983: tab. 6). Operculum slightly thickened, paucispiral, with low crest-like thickening at centre of inner face (Figs. 38, 39, 118; Bole, 1961: 62, 67, fig. 3A; Radoman, 1978: 30; 1983: 112; Bole, 1993: 10, fig. 2B). Body unpigmented (a few traces of pigment in wall of visceral sac); eye spots absent (Fig. 119). Male genitalia with prostate gland bulging well into pallial cavity; penis elongated, cylin- dro-conical, flat, slightly dilated then tapering near apex, with large non-glandular lobe on left side about 2/3 of penis length; penial duct zig-zagging through right portion of penis to open at penis tip (Figs. 120, 121; Bole, 1961: 62, fig. 3A; Radoman, 1978: 29, fig. 2C; 1983: 110, fig. 58C; Bole, 1993: 10, fig. 1C). Female genitalia with distal seminal recep- tacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle very long, slender; bursa copulatrix very large, oval to subtriangular, with long, slender duct entering bursa on anterior side; seminal groove run- ning along ventral side of capsule gland (Fig. 122; Radoman, 1978: 29, fig. 2A, B; 1983: 40, 110, fig. 58A, B). Bole (1993, fig. 1B), illus- trates female genitalia with a very small bursa copulatrix; this is due to a misidentification of a female of the Hauffenia species living in the same aquifer. Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 5-6 smaller denticles on both sides in de- creasing order of size; two basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 8-9 denti- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 137 FIGS. 117-122. Shell, operculum and anatomical details of Kerkia kusceri (Bole, 1961) from the spring of the Krka River, Krka, Slovenia, M. Bodon leg. 16.6.1985. Fig. 117: shell; Fig. 118: outer face (top centre), pro- file (top left and right) and inner face (below) of operculum; Fig. 119: body; Fig. 120: dorsal side of penis; Fig. 121: prostate gland, stomach, intestine and pallial organs of a male; Fig. 122: renal and pallial oviduct, in- testine and pallial organs of a female. Scale bar = 1 mm. 138 BODON, MANGANELLI & GIUSTI cles, central of which longer and larger; first marginal teeth with apical row of 12-16 denti- cles; second marginal teeth with rather long row of 14-18 denticles (Bole, 1961: 62, 67, fig. 3A; 1993: 10, fig. 2A). Stomach without posterior caecum; intes- tine with well developed, S-like bend on pallial wall (Figs. 121, 122; 1978: 29, fig. 2D; Rado- man, 1983: 40, fig. 58D). Osphradium variable in size, ovoid or elon- gated; ctenidium consisting of about 11-12 lamellae (Figs. 121, 122; Bole, 1961: 62, 67; 1993: fig. 1A). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1978: 29; 1983: 110). Taxonomy The genus Kerkia is characterized by: shell very small, valvatiform to planispiral; opercu- lum without peg, but with crest-like projection or thickening on inner face; penis with one simple lobe; female genitalia with distal semi- nal receptacle and very large, oval to subtri- angular, bursa copulatrix with anterior duct; central tooth with two pairs of basal cusps. It includes the type species and a recently de- scribed species: K. brezicensis Bodon & Cianfanelli, 1996, both from Slovenia. Kerkia brezicensis differs from K. kusceri in shell (flat spire), opercular (circular thickening on inner face), and anatomical (slender penial apex, smaller penial lobe) characters (Bodon & Cianfanelli, 1996). Lobaunia Haase, 1993 Lobaunia Haase, 1993: 98-99. Type Species: Lobaunia danubialis Haase, 1993, by original designation. Lobaunia danubialis Haase, 1993 Lobaunia danubialis Haase, 1993: 99-105, figs. 8B, 9-15. Type Locality: “Peilronr A 63 am Eber- schúttwasser im SE Wiens”, Austria. Type Material: holotype (NHMW 85027, fe- male) at the Naturhistorisches Museum Wien, Vienna, Austria; paratypes at the Naturhistorisches Museum Wien (NHMW 85028, 1 male: NHMW 85884, 31 shells) and in the Stojaspal collection (3230, 16 shells), Vienna, Austria (Haase, 1993). Material Examined —Pipe A89, Lobau, Wien, Austria, Pospisil leg. 24.1.1992, ex M. Haase collection (1 shell with dried soft parts, 8 shells). Description Shell very small, valvatiform-planispiral, thin, whitish, transparent when fresh; surface of protoconch malleated; spire almost flat, consisting of 2.25-2.75 rather rapidly growing convex whorls; last whorl dilated, descending slightly near aperture; umbilicus wide; aper- ture prosocline, oval; peristome complete, thin, slightly reflected only at columellar mar- gin (Fig. 123; Haase, 1993: 98-99, 104-105, fig. 9A-C; Boeters, 1998: 29, figs. H13-14). Dimensions: height = 0.50-0.84 mm; diame- ter = 0.86-1.31 mm (Haase, 1993: table 1). Operculum thin, yellow-orange, paucispiral, slightly thickened at centre, but without out- growth on inner face (Fig. 124; Haase, 1993: 98-99, figs. 10A, B). Body unpigmented (a few black spots on visceral sac level with stomach); eye spots absent (Haase, 1993: 98). Male genitalia with penis rather short, flat, tapering slightly near apex and ending in rather obtuse tip; penial duct zig-zagging through central portion of penis to open at penis tip; terminal portion of penial duct (im- mediately before opening) with very small stylet (Haase, 1993: 99, 103, Figs. 8B, 15; Boeters, 1998: 29, fig. H15). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle small but proportionally rather developed, arising from oviduct at end of loop; bursa copulatrix small, slightly larger than seminal receptacle, with very short duct entering on anterior side; according to Haase (1993), the seminal groove (“Ventralkanal”) is a distinct duct — possibly similar to the “spermathecal duct” de- scribed by Davis et al. (1982) in Spurwinkia salsa (Pilsbry, 1895)—completely separated from the lumen of the capsule gland, starting from proximal part of capsule gland and end- ing in the gonopore; the renal oviduct is sup- posed to end by entering the albumen gland through an opening (“Offnung fur Eizellen”); spermatozoa are supposed to enter the gono- pore, to run in the “spermathecal duct’, then enter the pallial oviduct and, finally, to reach renal oviduct (and hence seminal receptacle) through another opening (“Offnung fur Sper- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 139 matozoen”) connecting albumen gland di- rectly to renal oviduct (Haase, 1993: 98-99, 102, 104, figs. 8B, 12-14; Boeters, 1998: 29, fig. H16). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long and robust central denticle and 5 smaller denticles on both sides in decreasing order of size; 1-2 basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 10-11 denticles, central of which longer and larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 17-21 small denticles anteri- orly; second marginal teeth scraper-shaped, with long and slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 13-15 small denti- cles (Haase, 1993: 98-99, figs. 11A, B). Stomach without posterior caecum; intes- tine with well developed, S-like bend on pallial wall (Haase, 1993: 98-99, fig. 12). Osphradium unknown; ctenidium consist- ing of 8-9 lamellae (Haase, 1993: 99). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Haase, 1993: 99). Taxonomy Lobaunia is here considered a junior syn- onym of Hauffenia Pollonera, 1898 (see “Sta- tus and relationship of Hauffenia”). Haase (1993) described Lobaunia as a distinct genus based on the extremely peculiar struc- ture of the female genitalia (see description) reconstructed by serial sectioning. If these characters were true, this genus would be dis- tinct from all other European hydrobiids (val- vatiform and non-valvatiform). However, the fact that Lobaunia matches Hauffenia, Neo- horatia Schutt, 1961, Vrania Radoman, 1978, and Erythropomatiana Radoman, 1978, in male genital characters and also lives in an area where Hauffenia is widespread, invited prudence and suggested the need for more careful anatomical study. Asked for details of the anatomy of Lobaunia, Haase (personal communication, 3.2.1995) replied that he had new data suggesting that his reconstruction of the female genitalia of Lobaunia, based on only one specimen, might be incorrect and that Lobaunia might correspond to Hauffenia. Lobaunia included only the type species from Austria. Lyhnidia Hadzisée, 1959 Lyhnidia Hadzisce, 1959: 88. Type Species: Lyhnidia hadzii Hadzisée, 1959, by subsequent designation (Rado- man, 1963a). Lyhnidia hadzii Hadzisce, 1959 Lyhnidia hadzii HadZiSce, 1959: 90-93, figs. 27-29. Type Locality: “. . . lebt in den seichten Teilen des steinigen Litorals auf der óstlichen und südöstlichen Seite des Sees [Lake Ohrid] wie auch in seinem Sublitoral, námlich der Schalenzone und besonders der Zone der lebenden Dreissensien, Macedonia”. According to Radoman (1983: 116) the type locality is “Lake Ohrid, by Veli Dab”. Type Material: HadZisce (1959) did not give any information about the type material. Material Examined —Harbour of Trpezjca, Lake Ohrid, Macedo- nia, 34T DL 83, 5.1976, ex W. J. M. Maassen collection (2 shells with dried soft parts). Description Shell very small, valvatiform-globose coni- cal, pale whitish, waxen, transparent when fresh; surface of protoconch malleated; spire rather raised, conical, consisting of 3.25-3.5 rather rapidly growing convex whorls; last whorl large, slightly dilated, slightly descend- ing near aperture; umbilicus very small, slit- like; aperture prosocline, irregularly square; peristome complete, sinuous, with external margin concave above, convex at centre, concave again at border with the lower mar- gin, lower margin convex and slightly reflexed at border with columellar margin, columellar margin rather vertical, little thickened and re- flexed (Fig. 125; Hadzisce, 1959: 90-91, figs. 27-28; Radoman, 1963a: 77, fig. 5a; 1963b: 93, fig. 5a; 1983: 115-116, table 7, pl. 8, fig. 135). Dimensions: height = 0.92-1.15 mm; di- ameter = 0.84-1.08 mm, according to Rado- man, 1983: table 7 (height = 1.0-1.2 mm; di- ameter = 0.9-1.1 mm, according to HadZiSce, 1959). Operculum thick, yellowish red, paucispiral, 123 124 125 129 131 FIGS. 123-131. Shell and operculum of Hauffenia danubialis (Haase, 1993), from the pipe A89, Lobau, Wien, Austria, Pospisil leg. 24.1.1992, ex M. Haase collection (Figs. 123, 124), shell of Lyhnidia hadzii HadZisce, 1959, from the harbour of Trpezjca, Lake Ohrid, Macedonia, 5.1976, ex W. J. M. Maassen collec- tion (Fig. 125) and shell, operculum and anatomical details of Erythropomatiana verdica Radoman, 1978, ju- пог synonym of Hauffenia subpiscinalis (Kuácer, 1932), from spring on right bank of the Verd brook, near the Verd spring, Vrhnika, Slovenia, M. Bodon leg. 17.6.1985 (Figs. 126-131). Figs. 123, 125, 126: shell; Figs. 124, 127: outer face (left), profile (Fig. 124 centre; Fig. 127 right) and inner face (Fig. 124 right) of opercu- lum; Fig. 128: body of a male with pallial cavity open to show head and penis; Fig. 129: intestine and pallial organs of a male; Fig. 130: dorsal side (left) and ventral side (right) of penis; Fig. 131: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 141 without outgrowth at centre of inner face (HadZisce, 1959: 92). Body pigmented; eye spots present. Male genitalia with penis elongated, flat, ta- pering near apex, ending in rather pointed tip and with a small lobe on left side near apex (Hadzisce, 1959: 92, fig. 28; Radoman, 1963a: 77, fig. 13a; 1963b: 93, fig. 13a; 1983: 116). Hadzisce (1959, fig. 29) and Radoman (1963a: fig. 13a; 1963b: fig. 13a) show a dis- tinct penial lobe; however, Radoman’s (1983: 116) diagnosis of the genus does not mention this character: “Penis . . . smooth (without any outgrowth)”; the diagnosis is based, rather, on L. gjorgjevici, which has a penial tip without a distinct lobe. Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle rather well developed; bursa copulatrix large, kid- ney-shaped, with rather short, wide duct en- tering bursa on anterior side; seminal groove running along ventral side of capsule gland (Radoman, 1963a: 77, fig. 13a; 1963b: 93, fig. 13a; 1973a: 6, 10; 1983: 40, 114, 116). Rado- man’s diagnosis (1983: 116) described a bursa copulatrix “very small, hardly visible be- hind the oviduct loop, with a long duct (fig. 63)”, but it was based on L. gjorgjevici, not on L. hadzii. Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 4 smaller denticles on both sides in de- creasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with about 8-9 denticles, central of which longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of about 23 small denticles anteriorly; second marginal teeth scraper- shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of very small denti- cles (HadziSce, 1959: 92, fig. 29; Rado- man, 1963a, fig. 17b; 1963b, fig. 17b; 1983: 114). Stomach without posterior caecum; intes- tine unknown (Radoman, 1963a, fig. 13a; 1963b, fig. 13a; 1973a: 6; 1983: 40, 116). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and slightly shorter pleuro-suboe- sophageal connectives (Radoman, 1973a: 6; 1983: 116). Taxonomy Lyhnidia is here considered a distinct genus. However, some of its anatomical de- tails are unknown, and its relationships to other Balkan genera require further study. Lyhnidia is characterized by: shell very small, valvatiform-globose conical; operculum with- out peg; penis with one simple lobe; female genitalia with proximal seminal receptacle and large, kidney-shaped bursa copulatrix with anterior duct; central tooth with one pair of basal cusps. Apparently overlooking his earlier type des- ignation (as in the case of Karevia), Radoman (1967) erroneously reported Lyhnidia gjorgje- ис! Hadzisce, 1959, as a type species of Ly- hnidia. Radoman (1983) included four other spe- cies from Lake Ohrid in Lyhnidia: L. gjorgjevici Hadzisce, 1959, L. karamaniHadzisée, 1959, L. stankovici Hadzisce, 1959, L. sublitoralis Radoman, 1967, distinguished on the basis of few shell characters. Neohoratia Schutt, 1961a Neohoratia Schutt, 1961a: 71-72, as sub- genus of Horatia. Type Species: Valvata (?) subpiscinalis Kuscer, 1932, by original designation. Neohoratia subpiscinalis (Kuscer, 1932) Valvata (?) subpiscinalis KuScer, 1932: 51- 53, pled: 16. 1 Type Locality: “Der Rak-Bach unweit der ju- goslavisch-italienischen Grenze”, Slove- nia. Type Material: holotype (2001a) in the KuScer collection, Ljubljana, Slovenia (Kuscer, 1932). Erythropomatiana verdica Radoman, 1978: 36; РЁ: 5, 65.20.21. Type Locality: “Quelle im Bett des Flússchens neben dem Dorf Verd, unweit von Vrhnika”, Slovenia. Type Material: holotype and two paratypes (SMF 249615) are at the Senckenberg- Museum, Frankfurt am Main, Germany (Jovanovic, 1991). Material Examined Bole (1967a) studied the anatomy of speci- mens from the subterranean waters of the Ljubljanica basin, Slovenia. The specimens 142 BODON, MANGANELLI & GIUSTI FIGS. 132-140. Shell, operculum and anatomical details of Hauffenia subpiscinalis (Kuscer, 1932) from the “ZelSke Jame” caves, $. 576, near Rakek (Slovenia), 3.3.1966, ex Е. Velkovrh collection (Figs. 132-136, 138, 140) and from the spring Obrh, Gorenje Jezero, Cerknica, Slovenia, M. Bodon leg. 19.6.1985 (Figs. 137-139). Fig. 132: shell; Fig. 133: body of a male with pallial cavity open to show head and penis; Fig. 134: outer face (left) and profile (right) of operculum; Fig. 135: renal and pallial oviduct, intestine and pallial or- gans of a female; Figs. 136-137: renal and pallial oviduct of two females; Fig. 138: prostate gland, stomach, intestine and pallial organs of a male; Fig. 139: dorsal (left), right (centre) and ventral side (right) of penis; Fig. 140: dorsal side of penis of two males. Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 143 examined for the present study were from the following sites (the first two sites were at the spring and caves which feed the Rak River, so they belong to the type locality): _“Zelëke Jame” caves, $. 576, near Rakek, Slovenia, 33T VL 46, 3.3.1966, ex F. Velkovrh collection (2 males, 1 female); M. Buda Dancevich & F. Stoch leg. 1.7.1993 (2 shells). _ —Kotla spring, Rakov Skocjan. Plentiful karstic spring flooding into the Rak River, Slovenia, 33T VL 46, M. Bodon leg. 18.6.1985 (1 female, many shells); M. Buda Dancevich & F. Stoch leg. 1.7.1993 (many shells). —Obrh spring, Gorenje Jezero, Cerknica. Plentiful karstic spring in the CerkniSko lake basin (the lake water floods into sinkholes and feeds the Rak River), Slovenia, 33T VL 56, M. Bodon leg. 19.6.1985 (2 males, 1 female, 15 shells). —Spring of Zerovniscica, Zerovnica, Cerk- nica, Slovenia, 33T VL 57, F. Gasparo & F. Stoch leg. 14.10.1994 (1 male). —“Planinska Jama” cave, $. 748, Planina, Slovenia, 33T VL 47, F. Gasparo & F. Stoch leg. 5.6.1993 (1 male, 4 shells). —Mocilnik springs, Vrhnika; plentiful karstic springs which feed the Ljubljanica River, Slovenia, 33T VL 49, M. Bodon leg. 17.6.1885 (2 females, many shells). —Spring on right bank of Verd brook, near the Verd spring, Vrhnika, Slovenia, 33T VL 49, M. Bodon leg. 17.6.1985 (2 males, 1 female, many shells) (type locality of Erythropomatiana verdica). —Ukovnik cave, S. 1165, Spodnja Idrija, Idri- jca valley, Slovenia, 33T VM 20, F. Gas- paro & F. Stoch leg. 5.6.1993 (1 female). —“Draga pri Ponikvah” cave, S. 972, Ponikve, Stanjel, Slovenia, 33T VL 17, $. Dolce & F. Stoch leg. 19.7.1994 (1 male, 1 shell). —Springs of the Timavo River, S. Giovanni al Timavo, Duino-Aurisina, Trieste, Friuli- Venetia Julia, Italy, 33T UL 9071, M. Bodon leg. 31.3.1991 (1 female, many shells) (Pezzoli, 1988a, cited as /slamia (?) sp.). —Cave of Trebiciano, no. 17 VG, Trieste, Friuli-Venetia Julia, Italy, 33T VL 0959, S. Dolce & F. Stoch leg. 8.12.1991 (1 juv. specimen, 4 shells), 26.1.1992 (2 males, 1 female, 2 spec. and 7 juv. spec.), F. Stoch leg. 2.7.1996 (10 shells). — Spring in the square of Bagnoli della Rosan- dra which feeds a basin, Trieste, Friuli- Venetia Julia, Italy, 33T VL 1151, M. Bodon, M. M. Giovannelli & F. Stoch leg. 29.3.1991 (2 juv. spec., many shells). For other localities in Italy where only con- chological material has been found see Pez- zoli (1988a) and Bodon & Giovannelli (1993). For other localities in Slovenia, see KuScer (1932), Bole (1970, 1979, 1985) and Bole & Slapnik (1997). Description Shell very small, valvatiform, thin, pale whitish, waxen, transparent when fresh; sur- face of protoconch malleated; spire generally well raised, consisting of 3.25-3.75 rapidly growing, convex whorls; last whorl dilated, more or less descending, often slightly de- tached near aperture; umbilicus rather narrow; aperture wide, prosocline, roundish; peris- tome complete, thin, slightly reflected only at columellar margin (Figs. 90, 126, 132; KuScer, 1932: 51-52, pl. 5, fig. 1; Schútt, 1961a: 71; Bole, 1970: 92, fig. 3A-G; Bole: 1979a, fig. 1(5a-5b); Maassen: 1975, pl. 27, figs. 1-3; Bole 8 Velkovrh, 1986: fig. 31; Hershler & Lon- gley, 1986: fig. 71). Dimensions: height = 1.3- 2.6 mm; diameter = 1.4-2.9 mm. Operculum thin, yellowish, paucispiral, slightly thickened, but lacking peg or any kind of outgrowth at centre of inner face (Figs. 41, 127, 134; Bole, 1967a: 84, fig. 3A5; 1993: 8, fig. 2B). Body unpigmented (sometimes a few traces of pigment in wall of visceral sac); eye spots absent (Figs. 128, 133). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather short, flat, with apex enlarged, blunt or slightly pointed; 1-3 small, knob-like lateral lobes on left side near apex; sometimes slightly raised pleat at centre of ventral side about 2/3 of penis length; penial duct zig-zagging through central portion of penis to open at penis tip; large, pyriform mass of refringent cells inside penis apex to right of penial duct; terminal part of penial duct (immediately before opening) with very small stylet (Figs. 100, 129-130, 138-140; Bole, 1967a: 84, fig. 3A4; 1993: 8, fig. 1C). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle small, sessile or with very short duct arising from oviduct level with end of loop; bursa copulatrix reduced, small, but longer than seminal re- 144 BODON, MANGANELLI & GIUSTI ceptacle, not dilated at apex, arising very close to point at which oviduct enters albumen gland portion of pallial oviduct; seminal groove running along ventral side of capsule gland (Figs. 102, 103, 131, 135-137; Bole, 1967a: 84, fig. 3A3; 1993: 8, fig. 1B). Histo- logical study of female genitalia gave results in line with those described for H. tellinii. Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 5-6 smaller denticles on both sides in de- creasing order of size; 2-3 basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 9-13 denticles, central of which longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 19-25 small denticles anteriorly; sec- ond marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 10-12 very small denticles (Figs. 50-52; Bole, 1967a: 83, fig. 3A6; Hershler & Longley, 1986: fig. 28D; Bole, 1993: 8, fig. 2A). Stomach without posterior caecum; intes- tine with well developed, S-like bend on pallial wall (Figs. 129, 131, 135, 139; Bole, 1967a: 83). Osphradium variable in size, elongated, oval or kidney-shaped; ctenidium consisting of 9-16 lamellae (Figs. 129, 131, 135, 139; Bole, 1967a: 83, fig. 3A1; 1993: 8, fig. 1A). Nervous system with very short connec- tives (Bole, 1967a: 84). Taxonomy Neohoratia is here considered a junior syn- onym of Hauffenia Pollonera, 1898 (see “The status and relationships of Hauffenia”). Schütt (1961a) introduced Neohoratia, as subgenus of Horatia, for Valvata subpiscinalis KuScer, 1932. Subsequently, Neohoratia was re- garded as a subgenus of Hauffenia by Boeters (1974), completely overlooked by Radoman (1978, 1983), and ranked as a full genus by Bole 8 Velkovrh (1986), Boeters (1988, 1998) and Bole (1993). Erythropomatiana verdica Radoman, 1978, is here recognized as junior synonym of Neo- horatia subpiscinalis (see taxonomic remarks to H. subpiscinalis, in the section on Hauffenia species). Many nominal species from different south European localities (from Greece to Spain) have been assigned to Neohoratia (some- times regarded as a subgenus of Hauffenia or Horatia, sometimes as a distinct genus) by Schutt (1962, 1980), Bernasconi (1975), Boe- ters (1981, 1988, 1998), Gittenberger (1982), Vidal-Abarca & Suarez (1985), Boeters & Ro- lan (1988), Bech (1990), Hinz et al. (1994), Ramos et al. (1992) and Rolan (1997a, b). All were assigned to Neohoratia on the basis of shell characters, only in few cases followed by anatomical study. All are in need of revision (see below). Ohridohauffenia Hadzisce 1959 Ohridohauffenia Hadzisce, 1959: 74, as sub- genus of Ohridohoratia. Type Species: Ohridohoratia (Ohridohauffe- nia) gjorgjevici Hadzisce, 1959, by mono- typy. Ohridohoratia gjorgjevici is a junior synonym of Pseudamnicola depressa Radoman, 1957, according to Radoman (1963a, b, 1983). Ohridohauffenia depressa (Radoman, 1957) Pseudamnicola depressa Radoman, 1957: 88-91, figs. 2, 5, 7B, E. Type Locality: “. . . su uz istoënu obalu Ohrid- skog jezera, ро obalskom kamenju, pocev od izvora Veli Dab pa do pocetka peskovite juzne obale jezera, prema Sv. Naumi”, Lake Ohrid, Macedonia. Accord- ing to Radoman (1983: 75) the type lo- cality is “Lake Ohrid, on the bank stones by Veli Dab”. Type Material: lectotype (BEO 142, shell) is at Prirodnjacki Muzej u Beograd (Jo- vanovic, 1991). Ohridohoratia (Ohridohauffenia) gjorgjevici Hadzisce, 1959: 75-76, figs. 12, 13. Type Locality: “. . . die ich an seichten Stellen des steinigen Litorals gefunden habe . . .”, Lake Ohrid, Macedonia. Type Material: Hadzisce (1959) did not give any information about the type material. Description Shell very small, valvatiform, depressed; microsculpture of protoconch unknown; spire rather depressed, consisting of 3-3.25 rapidly growing, slightly convex whorls; last whorl large, rather dilated, with distinct keel at pe- riphery; umbilicus moderately wide; aperture prosocline, pyriform, distinctly angled at cen- tre of external margin, with external margin REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 145 slightly concave immediately above and below keel; peristome complete, thin, slightly reflected only at columellar margin (Rado- man, 1957: 88, fig. 2; HadziSce, 1959: 75-76, fig. 12, as Ohridohoratia (Ohridohauffenia) gjorgjevici; Radoman, 1963a: fig. 4a; 1963b: fig. 4a; 1983: 75, pl. 5, fig. 69; Jovanovic, 1991: pl. 5, fig. 35). Dimensions: height = 0.81-1.04 mm; diameter = 1.25-1.53 mm (Radoman, 1983: 75, table 5). Operculum thin, yellowish red, paucispiral, slightly thickened, without outgrowth on inner face (HadZisce, 1959: 76, as Ohridohoratia (Ohridohauffenia) gjorgjevici; Radoman, 1983: 75). Body pigmented; eye spots present. Male genitalia with penis large, flat with long conical tip, slightly obtuse apex, and one, more or less evident, sometimes pointed lobe on left side about 2/3 of penis length (Rado- man, 1957: 91, fig. 7E; Hadzisce, 1959: 76, fig. 12, as Ohridohoratia (Ohridohauffenia) gjorgjevici; Radoman, 1963: fig. 12a; 1963b: fig. 12a; 1983: 75, fig. 37). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal seminal receptacle much larger than distal, which is rudimentary; bursa copulatrix large, kidney-shaped, with long, slender duct entering bursa on anterior side; seminal groove running along ventral side of capsule gland (Radoman, 1957: 90-91, fig. 7B; 1963a: fig. 12a; 1963b: fig. 12a; 1973a: 6; 1983: 75: fig. 37). Radula with central tooth trapezoidal, its cutting edge with 9-11 denticles, central longer; long lateral wings and basal tongue; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their ante- rior margin with 8-11 denticles, central of which longer, larger; first marginal teeth rake- shaped, with long lateral wing and elongated cutting edge with long row of 14-21 small denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cut- ting edge carrying rather long row of nu- merous, very small denticles (Radoman, 1957: 90, fig. 5; HadZiSce, 1959: 76, fig. 13, as Ohridohoratia (Ohridohauffenia) gjorgje- Vici). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6; 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and slightly shorter pleuro-suboe- sophageal connectives (Radoman, 1973a: 6; 1983: 75). Taxonomy Ohridohauffenia is here considered a dis- tinct genus, but its relationships to Ohrigocea Hadziscée, 1959, and possibly also Karevia Hadzisce, 1959, need to be clarified. Ohrido- hauffenia is characterized by: shell very small, valvatiform, sometimes carinate; operculum without peg; penis with one simple lobe; fe- male genitalia with two seminal receptacles, proximal larger than distal (which is rudimen- tary), and large, kidney-shaped bursa copula- trix with anterior duct; central tooth with one pair of basal cusps. Rotondia Radoman, 1964 (pp. 108, 109; type species: Pseudamnicola (Rotondia) ro- tonda Radoman, 1964, by original designa- tion), is regarded as a junior synonym of Ohri- dohauffenia Hadzisce, 1959, by Radoman (1983). Rotondia was established without a description or definition, but only by combining it with some available nominal species; it is therefore not available (ICZN, 1999: Art. 13.6). Likewise, Naumia Radoman, 1973a, regarded by Radoman (1983) as a junior synonym of Ohridohauffenia, is not available. In fact, it was established by Radoman (1973a: 8, for Pseudamnicola (Rotondia) st. naumi [sic] Ra- doman, 1964) without a description or defini- tion, but only by combining it with an available species group name. After 1930, this condition does not make a generic name available (ICZN, 1999: Art. 13.6). Radoman (1983) includes five other species from Lake Ohrid and nearby springs in Ohridohauffenia: O. sublitoralis (Radoman, 1963a), O. rotonda (Radoman, 1964), O. min- uta (Radoman, 1955), O. drimica (Radoman, 1964), and O. sanctinaumi (Radoman, 1964). Ohrigocea Hadzisce, 1959 Ohrigocea Hadzisce, 1959: 76-77. Type Species: Ohrigocea samuili Hadzisce, 1959, by subsequent designation (Rado- man, 1963a). Ohrigocea samuili Hadzisce, 1959 Ohrigocea (Ohrigocea) samuili Hadzisce, 1959: 77-79, figs. 14, 15. Type Locality: “lebt in den seichten Zonen des steinigen Litorals auf der nord-óstlichen, óstlichen und sud-óstlichen Seite des 150 156 FIGS. 141-156. Shell and operculum of Ohrigocea samuili HadZiSce, 1959, from H. Gorica, Lake Ohrid, Macedonia, 5.1975, ex W. J. M. Maassen collection (Figs. 141, 142); shell, operculum and anatomical de- tails of Pezzolia radapalladis Bodon & Giusti, 1986, from the spring inside the Rio di Tonnego, Ponte della Vittoria, Rapallo, Genova, Liguria, Italy, M. Bodon leg. 16.5.1981, 4.10.1981, 6.11.1982 (Figs. 143, 144, 148-151) and from the spring on the right bank of the Bana Stream, below the tank of the aqueduct of Bana, Camogli, Genova, Liguria, Italy, M. Bodon leg. 26.1.1992 (Figs. 147, 152); anatomical details of Pezzolia sp. 2 from alluvial spring on bed of the Canate Stream near the cave of Cavassola, Genova, Liguria, Italy, 32T NQ 0222, M. Bodon leg. 24.12.1983 (Figs. 145, 155) and of Pezzolia sp. 1 from springs in the Nervi Stream valley, Genova, Liguria, Italy, 32T NQ 0315, M. Bodon leg. 15.1.1983, 32T NQ 0416, M. Bodon leg. 3.12.1983 (Figs. 146, 153, 154); shell of Prespolitorea valvataeformis Radoman, 1973a, from Lake Prespa, 3 Km north of Otesevo, Macedonia, ex W. J. M. Maassen collection (Fig. 156). Figs. 141, 143, 156: shell; Figs. 142, 144: outer face (Fig. 142 left, Fig. 144), profile (Fig. 142 centre) and inner face (Fig. 142 right) of operculum; Figs. 145-147: renal and pallial oviduct of three females; Fig. 148: intestine and pallial organs of a male; Fig. 149: renal and pallial oviduct, intestine and pallial organs of a female; Fig. 150: head of a male; Figs. 151-155: dorsal side of penis of six males. Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 147 Sees [Lake Ohrid]”, Macedonia. Accord- ing to Radoman (1983: 79), the type lo- cality is “Lake Ohrid, on bank stones by Veli Dab. Spread along all the rocky east lake bank, exclusively on stones”. Type Material: HadZisce (1959) did not give any information about the type material. Material Examined —H. Gorica, Lake Ohrid, Macedonia, 34T DL 84, 5. 1975, ex W. J. M. Maassen collec- tion (2 shells with operculum, 2 shells). Description Shell very small, valvatiform, sometimes depressed; microsculpture of protoconch un- known; spire slightly raised, consisting of 2.75-3 rather rapidly growing, convex whorls; last whorl, large, dilated, more or less de- scending near aperture; umbilicus wide; aper- ture prosocline, ovoid to pyriform, moderately angled at its upper vertex, its external margin slightly concave at upper half; peristome com- plete, thin, slightly reflected only at columellar margin (Fig. 141; Hadzisce, 1959: 77-78, fig. 14; Radoman, 1963a: fig. 6a; 1963b: fig. 6a; 1983: 79, pl. 5, fig. 74). Dimensions: height = 0.65-0.80 mm; diameter = 1.08-1.20 mm (Radoman, 1983: table 5). Operculum rather thick, yellowish red, pau- cispiral, thicker at centre of inner face, without peg (Fig. 142; Hadzisce, 1959: 78). Body pigmentation not described; eye spots present (Hadzisce, 1959: fig. 14) Male genitalia with penis rather elongated, conical, moderately pointed, and with one, rather evident, pointed lobe on left side about 2/3 of penis length (HadzZisce, 1959: 79, fig. 14; Radoman, 1963a: 78, fig. 14a; 1963b: 94, fig. 14a; 1983: 78, fig. 39). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal seminal receptacle larger than distal, which is reduced; bursa copulatrix large, kidney-shaped, with long, slender duct entering bursa on anterior side; seminal groove running along ventral side of capsule gland (Radoman, 1963a: 78, fig. 14a; 1963b: 94, fig. 14a; 1973a: 6; 1983: 40, 78, fig. 39). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long, robust central denti- cle and 4 smaller denticles on both sides in decreasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, their anterior margin with about 9 denticles; first marginal teeth rake-shaped, with row of about 20 denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cut- ting edge carrying row of about 20 denticles (Hadzisce, 1959: 78-79, fig. 15; Radoman, 1963a: 90, fig. 17c; 1963b: 109, fig. 17c). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6; 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and slightly shorter pleuro-suboe- sophageal connectives (Radoman, 1973a: 6; 1983: 78). Taxonomy Ohrigocea is here considered a distinct genus, but its relationships to Ohridohauffenia Hadzisce, 1959, and possibly also Karevia Hadzisce, 1959, need to be clarified. Ohrigo- cea is Characterized by: shell very small, val- vatiform, depressed, sometimes carinate; op- erculum without peg; penis with one simple lobe; female genitalia with two seminal recep- tacles, proximal larger than distal, which is rudimentary, and large, kidney-shaped bursa copulatrix with anterior duct; central tooth with one pair of basal cusps. Ohridosturanya Radoman, 1973a, re- garded by Radoman (1983) as a junior syn- onym of Ohrigocea, is not available. In fact, it was established by Radoman (1973b: 8; for Horatia (Hauffenia) stankovici Hadzisée, 1959) without a description or definition, but only by combining it with an available species- group name. After 1930, this condition does not make a generic name available (ICZN, 1999: Art. 13.6). Radoman (1983: 78-80), assigned three other species to Ohrigocea: О. karevi Hadzisce, 1959, O. miladinovorum Hadzisée, 1959, and O. stankovici (HadzZisce, 1959). All are distinguished mainly by shell characters and live, together with O. samuili, in the Lake Ohrid. Pezzolia Bodon & Giusti, 1986 Pezzolia Bodon 4 Giusti, 1986: 62-63. Type Species: Pezzolia radapalladis Bodon & Giusti, 1986, by original designation. 148 BODON, MANGANELLI 8 GIUSTI Pezzolia radapalladis Bodon & Giusti, 1986 Pezzolia radapalladis Bodon € Giusti, 1986: 63-66, table 1, figs. 1A-G, pl. 1, figs. A- D, pl. 2, figs. A-D. Type Locality: “spring inside the Rio di Ton- nego, close to the Ponte della Vittoria, Rapallo, Genova. 3°14’34”М; 44°22’ 14”E; 30 m a.s.l. Square on the map of the Military Geographic Institute: 83 II SO, Rapallo”, Italy. Type Material: the holotype (SMF 305486, shell) is at the Senckenberg-Museum, Frankfurt am Main, Germany; paratypes (shells and preserved specimens) are at the Museo Civico di Storia Naturale G. Doria, Genoa, Italy, and in the Giusti (Siena, Italy), Bodon (Genoa, Italy), and Pezzoli (Milan, Italy) collections (Giusti & Pezzoli, 1981). Material Examined —Spring inside the Rio di Tonnego, Ponte della Vittoria, Rapallo, Genova, Liguria, Italy, 32T МО 1613, M. Bodon leg. 16.12.1979, 31.12.1979, 29.11.1980, 3.12.1980, 4.4.1981, 3.4.1981, 16.5.1981, 4.10.1981, 6.11.1982 (8 males, 7 females, some specimens, many shells). —Spring at Le Rocche, Casalino, Rapallo, Genova, Liguria, Italy, 32T NQ 1512, S. Gaiter leg. 28.8.1991 (1 female, many shells). —Spring on right bank of the Bana Stream, below the tank of the Bana aqueduct, Camogli, Genova, Liguria, Italy, 32T NQ 1410, M. Bodon leg. 26.1.1992, 12.4.1992 (3 males, 5 females, some specimens, many shells). —Cave Valdettaro no. 129 Li, Rapallo, Ra- pallo, Genova, Liguria, Italy, 32T NQ 1710, M. Bodon leg. 6.11.1992 (1 male, 3 females, 11 shells). —Spring on left bank of the S. Francesco Stream, Il Campo, Rapallo, Genova, Li- guria, Italy, 32T NQ 1913, M. Bodon leg. 31.3.1996, 15.12.1996 (2 males, 1 fe- male, many shells). —Spring on right bank of the Recco Stream, below the church of Avegno, Genova, Liguria, Italy, 32T NQ 1315, M. Bodon leg. 10.9.1994, 12.11.1994, 19.11.1996 (1 male, 3 females, a few shells). For other localities where only shells have been collected see Pezzoli, 1988a. Description Shell very small, valvatiform, thin, whitish, waxen, transparent when fresh; surface of protoconch malleated; surface of teleoconch with thin growth-lines; spire raised, rarely de- pressed, planispiral, consisting of 2.5-3.25 rather rapidly growing convex whorls; last whorl large, slightly dilated, descending near aperture; umbilicus wide; aperture prosocline, oval or slightly pyriform; peristome complete, thin, slightly reflected only at columellar mar- gin (Figs. 91, 143; Bodon & Giusti, 1986: 62-64, pl. 1, figs. A-D). Dimensions: height = 0.58-1.05 mm; diameter = 0.95-1.37 mm. Operculum thin, pale yellowish, paucispiral, slightly thicker, but without traces of outgrowth at centre of inner face (Fig. 144; Bodon & Giusti, 1986: 62, 64, fig. 1C). Body unpigmented (a few traces of pigment in wall of visceral sac); eye spots absent (Fig. 150; Bodon & Giusti, 1986; 62, 64, fig. 1A, B). Male genitalia with penis from short to elon- gated, flat, without lobes; penis terminal por- tion widening slightly before ending in short, pointed, conical-flat apex; penial duct zig-zag- ging through right portion of penis to open at penis tip (Figs. 151, 152; Bodon & Giusti, 1986: 62, 64, fig. 1B, G, table 1). Female genitalia with two seminal recepta- cles and bursa copulatrix (when present) aris- ing from distal renal oviduct; proximal seminal receptacle small, sometimes slightly smaller than distal; bursa copulatrix very reduced or absent, with very short duct so as to appear equal in size to distal seminal receptacle or sometimes smaller; seminal groove running along ventral side of capsule gland (Figs. 147, 149; Bodon & Giusti, 1986: 62, 64, fig. 1E, F, table 1). Radula with central tooth trapezoidal with long lateral wings and basal tongue; anterior margin with 15-19 denticles, central of which longer, larger; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 13-18 denticles, central of which longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 11-16 small denticles anteriorly; second marginal teeth scraper-shaped, with long and slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of very small denticles (Bodon & Giusti, 1986: 62, 64, pl. 2, figs. A-D). Stomach without posterior caecum; intes- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 149 tine with well-developed S-like bend on pallial wall (Figs. 148-149; Bodon & Giusti, 1986: 62, 64, fig. 1D, E). Osphradium oval or reniform; ctenidium ab- sent (Figs. 148-149; Bodon 8 Giusti, 1986: 62, 64, Fig. 1D, E). Nervous system unknown. Taxonomy Pezzolia is characterized by: shell very small, valvatiform; operculum without peg; penis with or without glandular lobe/s; female genitalia with two seminal receptacles, proxi- mal equal or smaller than distal, and bursa copulatrix very reduced or absent; central tooth with one pair of basal cusps. In the only population of Pezzolia radapal- ladis studied by Bodon & Giusti (1986), the bursa copulatrix was always present, but in other populations subsequently discovered, it may be extremely reduced or even absent (Fig. 147). Other populations of Pezzolia re- cently found in the subterranean waters of Liguria are distinct from Р radapalladis by virtue of conchological (more globose shell) and anatomical (penis with 1-2 glandular lobes) characters (Figs. 145, 146, 153-155). This may support the existence of more than one species of Pezzolia (Pezzoli, 1988a; Bodon et al., 1995b). Prespolitorea Radoman, 1983 Prespolitorea Radoman, 1983: 68. Type Species: Prespolitorea valvataeformis Radoman, 1983, by original designation. Prespolitorea valvataeformis Radoman, 1973a Prespolitoralia [sic] valvataeformis Radoman, 1973a: 20-21. Type Locality: “stony, south-west coast of Lake Prespa, on the stones in the shal- low coastal zone”, Macedonia. Type Material: lectotype (BEO 120, shell) at the Prirodnjacki Muzej u Beograd, to- gether with a paralectotype (BEO 121, shell) (Jovanovic, 1991). Material Examined —Lake Prespa, 3 km north of Otesevo, Mace- donia, 33T DL 94, ex W. J. M. Maassen collection (1 shell). Description Shell very small, valvatiform; surface of pro- toconch malleated; spire rather depressed, consisting of 3-3.5 rather rapidly growing, convex whorls; last whorl large, dilated, slightly descending near aperture; umbilicus moderately wide, sometimes partly covered by reflected columellar margin of peristome; aperture prosocline, irregularly ovoid, its ex- ternal margin sinuous, somewhat angled at periphery; peristome complete, thin, slightly thickened, reflected at columellar margin (Fig. 156; Radoman, 1973a: 20-21; 1983: 68, pl. 4, fig. 60; Jovanovic, 1991: pl. 4, fig. 28). Dimensions: height = 0.84-1.01 mm; di- ameter = 1.18-1.30 mm (Radoman, 1983: table 4). Operculum probably without outgrowth, though not specified. Body pigmented; eye spots present. Male genitalia with penis large, flat, apically extended in rather pointed conical tip, and with one small pointed lobe on left side about 2/3 of penis length (Radoman, 1973a: 21; 1983: 68, fig. 31). Female genitalia with two seminal recepta- cles and a bursa copulatrix; proximal seminal receptacle much larger than distal, which is rudimentary; bursa copulatrix large, globose, with long, slender duct entering bursa on an- terior side; seminal groove running along ven- tral side of capsule gland (Radoman, 1973a: 6, 21; 1983: 40, 68, fig. 31). Radula with central tooth with one pair of basal cusps; other details unknown (Rado- man, 1983: 40). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6; 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and shorter pleuro-suboesopha- geal connectives (Radoman, 1973a: 6; 1983: 68). Taxonomy As Radoman (1973a: 7, 20-21) established Prespolitorea without a description or defini- tion, but only by combining it with two new species (one of which he designated as type species), this nominal genus was not made available in 1973 (ICZN, 1999: Art. 13.6). On the contrary, the two new nominal species are available (ICZN, 1999: Art. 11.9.3). Prespoli- torea is here considered a distinct genus. However, some of its anatomical details are 150 BODON, MANGANELLI & GIUSTI unknown, and its relationships to other Balkan genera (especially Daphniola Radoman, 1973, and Horatia Bourguignat, 1887) require further study. Prespolitorea is characterized by: shell very small, valvatiform; operculum without peg; penis with one simple lobe; fe- male genitalia with two seminal receptacles, proximal larger than distal (which is rudimen- tary), and large, globose bursa copulatrix with anterior duct. Radoman (1973a: 21) included in Prespoli- torea the type species, endemic to Lake Prespa, and another species from Lake Malo, south of Lake Prespa, Albania, P. malapres- pensis Radoman, 1973a, distinguished from P. valvataeformis by shell shape. Pseudohoratia Radoman, 1967 Pseudohoratia Radoman, 1967: 149-151. Type Species: Valvata ochridana Polinski, 1929, by original designation. Pseudohoratia ochridana (Polinski, 1929) Valvata (Atropidina) ochridana Polinski, 1929: 136-137. Type Locality: “[Lake Ohrid], Macedonia. Ac- cording to Radoman (1983: 115): “Lake Ohrid, in the Chara zone in the Ohrid gulf”. Type Material: Polinski (1929) did not give any information about the type material. Material Examined —Lake Ohrid, Macedonia, 34T DL, ex P. Radoman collection (2 males, 1 female). Description Shell very small, valvatiform, thin, yellow- ish, glassy, transparent when fresh; surface of protoconch malleated; spire raised, consist- ing of 3-3.25 rather rapidly, regularly growing, convex whorls; last whorl large, slightly di- lated, more or less descending near aperture; umbilicus from small to wide; aperture proso- cline, roundish to oval; peristome complete, slightly thickened, slightly reflected only at columellar margin (Figs. 92, 157; Polinski, 1929: 136-137; 1932: 617, pl. 7, fig. 4; Rado- man, 1953: 64-66, figs. 1-3, 8, table 1; 1955: 60, table 4, pl. 5, figs. 9-11; 1967: 149, fig. 1; 1983: 114, pl. 8, fig. 131). Dimensions: height = 0.94-1.30 mm; diameter = 1.00-1.39 mm (Radoman, 1983: table 7). Operculum slightly thickened, reddish yel- low, paucispiral, with small, not or slightly api- cally dilated peg at centre of inner face (Figs. 40, 159; Radoman, 1953: 66, fig. 4; 1955: 57-58; 1967: 149, fig. 1; 1983: 114) Body unpigmented; eye spots present (Fig. 158). Male genitalia with prostate gland bulging well into pallial cavity; penis cylindrical, elon- gated, tapering near apex, ending in rather pointed tip, with small, non-glandular lobe on left side about 2/3 of penis length; penial duct zig-zagging through right or central (near tip) portion of penis to open at penis tip (Figs. 160, 161; Radoman, 1955: 43-44, fig. 99; 1967: fig. 2c; 1983: 114, fig. 61D). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle small but proportionally rather developed, bent to adhere to oviduct; bursa copulatrix very small, with long, slender duct entering bursa on an- terior side; seminal groove running along ven- tral side of capsule gland (Fig. 162; Radoman, 1955: 36, 38-39, figs. 82, 83, 86-97; 1967: 150, fig. 2d; 1973a: 6, 10; 1983: 40, 114, fig. 61C). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long, robust central denti- cle and 3-4 smaller denticles on both sides in decreasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 9-11 den- ticles, central of which longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 15-18 small denticles anteriorly; sec- ond marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of very small denticles (Radoman, 1955: 12-13, fig. 17, table 2; 1967: 150, fig. 2a; 1983: 114, fig. 61A). Stomach without posterior caecum; intes- tine without bend on pallial wall (Figs. 161, 162; Radoman, 1955: 21-24, figs. 34, 35; 1973a: 6; 1983: 40). Osphradium variable in size, elongated, el- liptical; ctenidium consisting of 7-9 lamellae (Figs. 161, 162; Radoman, 1955: 11, table 1). Nervous system with long pleuro-supraoe- sophageal and shorter pleuro-suboesoph- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 157 FIGS. 157-170. Shell, operculum and anatomical details of Pseudohoratia ochridana (Polinski, 1929) from Lake Ohrid, Macedonia, ex P. Radoman collection (Figs. 157-162) and of Hauffenia wagneri (Kuácer, 1928) from the spring of the “Vranja реб” cave, Bostanj, Sevnica, Kráko, Slovenia, M. Bodon leg. 14.6.1985 (Figs. 163-170). Figs. 157, 163-165: shells; Figs. 158, 168: body of a male with pallial cavity open to show head and penis; Figs. 159, 166: outer face (left), profile (centre; also extreme right in Fig. 166) and inner face (right) of operculum; Figs. 160, 169: dorsal side of penis of two males; Figs. 161, 170: prostate gland, stomach (ex- cluded in Fig. 170), intestine and pallial organs of a male; Figs. 162, 167: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar = 1 mm. 152 BODON, MANGANELLI & GIUSTI ageal connectives (Radoman, 1955: 45-46, fig. 123; 1967: 150, fig. 2b; 1973a: 6; 1983: 114, fig. 61B). Taxonomy Pseudohoratia is characterized by: shell very small, valvatiform; operculum with peg; penis with one simple lobe; female genitalia with proximal seminal receptacle and bursa copulatrix very small, but with long anterior duct; central tooth with one pair of basal cusps. Horatia polinskii Radoman, 1953 (pp. 64-67, figs. 1-4; type locality: “. . . lac d'Ohrid ... entre 20 et 120 m de profondeur . . .”) is a junior synonym of Pseudohoratia ochri- dana (Polinski, 1929) according to Radoman (1983). Radoman (1983: 115) included two more species from Lake Ohrid in Pseudohoratia: P. brusinae (Radoman, 1953) and Р lacustris (Radoman, 1964). They are distinguished by shell shape. Two out of 100 females of Р. brusinae had a very rudimentary distal semi- nal receptacle (Radoman, 1983: 115). Pseudoislamia Radoman, 1979 Pseudoislamia Radoman, 1979: 23. Type Species: Pseudoislamia balcanica Radoman, 1979, by original designation. Pseudoislamia balcanica Radoman, 1979 Pseudoislamia balcanica Radoman, 1979: 23-27, fig. 1, table 1, pl. 1, figs. 1, 2. Type Locality: “Trichonis Lake, along the stone northeast bank, near by Mirtia, Greece”. Type Material: the lectotype (BEO 171, shell) is at the Prirodnjacki Muzej u Beograd to- gether with a paralectotype (BEO 172, shell) (Jovanovic, 1991). Description Shell very small, valvatiform, depressed; microsculpture of protoconch unknown; spire depressed, consisting of 2.75-3.25 rather rapidly growing convex whorls; last whorl large, dilated; umbilicus rather wide, deep; aperture prosocline, irregularly oval due to sinuous contour (upper part of external mar- gin slightly concave; central part of external margin projected forwards, angled; lower margin convex); peristome complete, thin, slightly reflected only at columellar margin (Radoman, 1979: 23, 27, pl. 1, figs. 1, 2; 1983: 83, pl. 5, figs. 84-86; Jovanovic, 1991: pl. 6, fig. 43). Dimensions: height = 0.84-1.05 mm; diameter = 1.18-1.34 mm (Radoman, 1979: table 1; 1983: table 5). Operculum presumably without outgrowth though not specified. Body pigmentation not described; eye spots present (Radoman, 1979: fig. 1; 1983: fig. 44). Male genitalia with penis elongated, with one evident lobe on left side near apex (Rado- man, 1979: 23, 27, fig. 1; 1983: 83, fig. 44). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal seminal receptacle slightly larger than distal; bursa copulatrix small, with long, slender duct entering bursa on anterior side; seminal groove running along ventral side of capsule gland (Rado- man, 1979: 6, 23, 27, fig. 1; 1983: 40, 83, fig. 44). Radula with central tooth with one pair of basal cusps; other details unknown (Rado- man, 1979: 23; 1983: 40). Stomach without posterior caecum; intes- tine unknown (Radoman, 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and shorter pleuro-suboesoph- ageal connectives (Radoman, 1979: 23; 1983: 83). Taxonomy The genus Pseudoislamia is characterized by: shell very small, valvatiform, depressed; operculum without peg; penis with one simple lobe; female genitalia with two seminal recep- tacles, proximal sligtly larger than distal, and very small bursa copulatrix with long anterior duct; central tooth with two pairs of basal cusps. It includes only the type species which is endemic to Lake Trichonis, Greece (Rado- man, 1983). Sardohoratia Manganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998 Sardohoratia Manganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998: 51-53. Type Species: Sardohoratia sulcata Man- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 153 ganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998, by original designation. Sardohoratia sulcata Manganelli, Bodon, Cianfanelli, Talenti 8 Giusti, 1998 Sardohoratia sulcata Manganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998: 53-55, figs. 28-30, 34-37, 40-45, 53-57. Type Locality: “Plentiful karstic spring “Su Col- ogone” at San Giovanni, 100 m a.s.l. (Oliena, Nuoro). UTM references: 32T NK 4260”, Sardinia, Italy. Type Material: holotype (MZUF 11580, shell) and four paratypes (MZUF 11559, 11581, 4 shells) in the malacological collection of the Museo Zoologico “La Specola”, Florence, Italy; other paratypes (145 shells and 14 preserved specimens) are in the Bodon (Genova, Italy), Cianfanelli (Florence, Italy), Giusti (Siena, Italy), Maassen (Duivedrecht, Holland), Sosso (Genoa, Italy), and Talenti (Florence, Italy) collections (Manganelli et al., 1998). Material Examined —Plentiful karstic spring “Su Cologone”, San Giovanni, Oliena, Nuoro, Sardinia, Italy, 32T NK 4260, M. Bodon, F. Giusti & G. Manganelli leg. 22.11.1986 (44 shells), M. Bodon leg. 24.3.1989 (3 males, 4 fe- males, 7 specimens, many shells). Description Shell very small, valvatiform, rather robust, whitish, waxen; surface of protoconch mal- leated surface of teleoconch with deep, scat- tered spiral groves; spire raised, consisting of 2.25-3 rather rapidly growing convex whorls; last whorl slightly dilated, not or slightly descending near aperture; umbilicus rather small; aperture prosocline, ovoid; peri- stome complete, thickened, columellar mar- gin slightly reflected (Manganelli et al., 1998: 53-54, figs. 28-30, 34-37). Dimensions: height = 0.60-1.13 mm; diameter = 0.62-1.24 mm (Manganelli et al., 1998: table 1). Operculum very thin, pale yellow, paucispi- ral, slightly thicker at centre, lacking out- growth on inner face (Manganelli et al., 1998: 54, fig. 40). Body unpigmented; eye spots absent (Man- ganelli et al., 1998: 54, fig. 41). Male genitalia with penis rather elongated, slender, with pointed apex, without lobes; pe- nial duct zig-zagging through central portion of penis to open at penis tip (Manganelli et al., 1998: 54, figs. 41-43). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; seminal receptacles of equal size; proximal seminal receptacle bent out- wards near end of loop; bursa copulatrix large, oval, with rather long duct entering bursa on anterior side; seminal groove run- ning along ventral side of capsule gland (Man- ganelli et al., 1998: 54, figs. 44, 45). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long, robust central denti- cle and 5 smaller denticles on both sides in decreasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 9 denti- cles, central of which longer, larger; first mar- ginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 20-22 small denticles anteriorly; second mar- ginal teeth scraper-shaped, with long slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of ap- proximately 12-15 small denticles (Man- ganelli et al., 1998: 54, figs. 53-57). Stomach without posterior caecum; intes- tine with S-like bend on pallial wall (Man- ganelli et al., 1998: 54-55, figs. 42, 44). Osphradium oval; ctenidium absent (Man- ganelli et al., 1998: 54-55, figs. 42, 44). Nervous system unknown. Taxonomy The genus Sardohoratia is characterized by: shell very small, valvatiform; operculum without peg; penis without lobes; female gen- italia with two seminal receptacles equal in size and large, oval bursa copulatrix with an- terior duct; central tooth with one pair of basal cusps. Besides the type species, Sardohoratia in- cludes another species from Sardinia, S. is- lamioides Manganelli, Bodon, Cianfanelli, Ta- lenti & Giusti, 1998, which differs in a few characters. Strugia Radoman, 1973a Strugia Radoman, 1973a: 10. Type Species: Strugia ohridana Radoman, 1973a, by monotypy. 154 BODON, MANGANELLI & GIUSTI Strugia ohridana Radoman, 1973a Strugia ohridana Radoman, 1973a: 10, 25. Type Locality: “Cave from which Sum spring comes out, about 4 km west of town Struga, Macedonia”. Type Material: lectotype (BEO 262, shell) is at the Prirodnjacki Muzej u Beograd, to- gether with a paralectotype (BEO 263, shell) (Jovanovic, 1991). Description Shell very small, valvatiform; microsculp- ture of protoconch unknown; spire rather raised, consisting of 3.5-3.75 rather rapidly and regularly growing convex whorls; last whorl large, slightly dilated, more or less de- scending near aperture; umbilicus wide; aper- ture prosocline, roundish; peristome com- plete, slightly thickened, slightly reflected only at lower and columellar margin (Radoman, 1973a: 25; 1983: 118, pl. 8, fig. 139; Jo- vanovic, 1991: pl. 8, fig. 65). Dimensions: height = 1.50-1.60 mm; diameter = 1.60-1.68 mm (Radoman, 1983: table 7). Operculum reddish yellow, paucispiral, without outgrowth at centre of inner face (Radoman, 1973a: 25; 1983: 118). Body unpigmented; eye spots absent (Radoman, 1983: 119). Male genitalia with penis rather elongated, slightly dilated (at about 2/3 of penis length), then tapering to end in moderately pointed tip; small knob-like lobe on left side of penis level with subapical dilated portion (Radoman, 1983: 119, fig. 65). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle pro- portionally well developed, bent to adhere to oviduct; bursa copulatrix moderately large, sac-like or kidney-shaped, with rather long, slender duct entering bursa on anterior side; seminal groove running all along ventral side of capsule gland (Radoman, 1973a: 6, 25; 1983: 119, fig. 65). Radula with central tooth with one pair of basal cusps; other details unknown (Rado- man, 1983: 114). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6; 1983: 40) Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and shorter pleuro-suboesopha- geal connectives (Radoman, 1973a: 6; 1983: 40, 118). Taxonomy Although Radoman (1973a: 10, 25) estab- lished Strugia without a description or defini- tion, but only by combining it with a new species, this nominal genus is available (ICZN, 1999: Art. 13.4). Strugia is here considered a distinct genus, but its relationships to Lyhnidia Hadzisée, 1959, need to be clarified. Strugia is charac- terized by: shell very small, valvatiform; oper- culum without peg; penis with one simple lobe; female genitalia with proximal seminal receptacle and large, sac-like or kidney- shaped bursa copulatrix with anterior duct; central tooth with two pairs of basal cusps. A monotypic genus including only the type species, endemic to the Ohrid Basin. Vrania Radoman, 1978 Vrania Radoman, 1978: 35, as subgenus of Hauffenia. Type Species: Valvata wagneri Kuscer, 1928, by original designation. Vrania wagneri (KuSCer, 1928) Valvata wagneri Kuscer, 1928: 50, fig. 1. Type Locality: “Grotte Vranja реб bei Bostanj, 46°N, 1577'E, Slovenia. Type Material: Kuscer (1928) did not give any information about the type material. Material Examined —Spring of the “Vranja pec” cave, BoStanj, Sevnica, Kr$ko, Slovenia, 33T WL 29, M. Bodon leg. 14.6.1985 (2 males, 2 fe- males, 6 shells). —Spodnja KlevevSka Jama, $. 410, Smarjeta, Novo Mesto, Slovenia, 33T WL 18, F. Stoch leg. 16.6.1996 (1 female). Description Shell very small, markedly conical-valvati- form or valvatiform, thin, pale whitish, waxen, transparent when fresh; surface of proto- conch malleated; spire from moderately to well raised, consisting of 2.75-3.5 rather rapidly growing, convex whorls; last whorl di- lated, sometimes descending slightly near aperture; umbilicus moderately wide; aper- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 195 FIGS. 171-179. Radula of Hauffenia wagneri (Kuscer, 1928) from the spring of the “Vranja pec” cave, BoSanj, Sevnica, KrSko, Slovenia, M. Bodon leg. 14.6.1985 (Fig. 171), Fissuria raehlei (Schütt, 1980) from well no. G/143, Poros, Cephalonia, Greece, G. L. Pesce, D. Maggi 8 G. Silverii leg. 2.4.1978 (Figs. 172, 173), Islamia consolationis (Bernasconi, 1985) from the cave Biez-Airoux, Consolation-Maisionnettes, Doubs, France, M. Bodon 8 G. Manganelli leg. 13.6.1996 (Figs. 174-176) and Heraultia exilis (Paladilhe, 1867) from the spring of Lez River, Hérault, France, M. Bodon leg. 2.12.1984 (Figs. 177-179). Figs. 171, 175: half of central portion of radula; Figs. 172, 177: central teeth; Figs. 173, 179: lateral, inner and outer marginal teeth; Fig. 174: central and lateral teeth; Fig. 176: outer marginal tooth; Fig. 178: four lateral teeth. Scale bar = 5 um. 156 BODON, MANGANELLI & GIUSTI ture prosocline, roundish; peristome com- plete, thin, slightly reflected only at columellar margin (Figs. 93, 163-165; Kuscer, 1928: 50, fig. 1; Radoman, 1978: 35, pl. 4, figs. 16, 17; 1983: 122-123, pl. 9, fig. 145, table 7). Di- mensions: height = 0.71-1.14 mm; diameter = 0.95-1.60 mm. Operculum thin, yellowish, paucispiral, slightly thickened, with small, not apically di- lated but spiralized peg at centre of inner face (Figs. 42, 43, 166; Bole, 1970: 92, fig. 2B5; Radoman, 1978: 35, fig. 5F; 1983: 122, fig. 67F). Body unpigmented (a few traces of pigment in wall of visceral sac); eye spots absent (Fig. 168). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather short, flat, with apex blunt, and one rather evident, wide lobe on left side near apex; penial duct zig-zagging through central portion of penis to open at penis tip; large, roundish or pyriform mass of refringent cells present inside penis apex right of penial duct; terminal part of pe- nial duct (immediately before opening) with very small stylet (Figs. 169-170; Bole, 1970: 92, fig. 2B3). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle very small, sessile, or with very short duct arising from oviduct level with end of loop; bursa copulatrix reduced, small but longer than seminal receptacle, not or slightly dilated at apex, arising very close to where oviduct en- ters albumen gland portion of pallial oviduct; seminal groove running along ventral side of capsule gland (Fig. 167; Bole, 1970: 92, fig. 2B2). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 5-6 smaller denticles on both sides in de- creasing order of size; 1-2 basal cusps at point where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 10-12 denticles, central of which longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 21-24 small denticles anteriorly; sec- ond marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 14-19 very small denticles (Fig. 171; Bole, 1970: 92, fig. 2B4). Stomach without posterior caecum; intes- tine with well-developed, tightly coiled, S-like bend on pallial wall (Figs. 167, 170; Rado- man, 1978: 33; 1983: 40). Osphradium variable in size, oval or elon- gated, kidney-shaped; ctenidium consisting of 7-11 lamellae (Figs. 167, 170; Bole, 1970: 92, fig. 2B1). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1978: 33; 1983: 120). Taxonomy Vrania is here considered a junior synonym of Hauffenia Pollonera, 1898 (see “The status and relationship of Hauffenia”). Radoman (1978) introduced this taxon, as subgenus of Hauffenia, for Valvata wagneri KuScer, 1928. Vrania was confirmed as subgenus of Hauffe- nia by Radoman (1983) and Bole & Velkrovh (1986). On the contrary, Bole (1993) and Haase (1993) regarded it as a junior synonym of Hauffenia. Vrania included only the type species. Zaumia Radoman, 1983 Zaumia Radoman, 1983: 119. Type Species: Horatia kusceri Hadzisce, 1959, by original designation. Zaumia kusceri (HadZisce, 1959) Horatia kusceri Hadzisce, 1959: 65-66, figs. 4, 5. Type Locality: “lebt in den Quellen des Klosters st. Naum am Súdende des Sees und an den sechten, steinigen Teilen der Litoralregion auf der Ostseite des Sees”, Ohrid Basin, Macedonia. According to Radoman (1983: 120), the type locality is “springs by Sveti Naum, near the south bank of Lake Ohrid”. Type Material: Hadzisce (1959) did not give any information about the type material. Description Shell very small, valvatiform or conical- ovoid, thin, glassy, transparent when fresh; mi- crosculpture of protoconch unknown; spire well raised, consisting of 3.25-3.5 rather rapidly growing convex whorls; last whorl large, dilated, descending slightly near aper- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 157 ture; umbilicus small, hole-like; aperture prosocline, roundish to ovoid; peristome complete, thin, slightly thickened, slightly re- flected only at lower and columellar margin (HadZiSce, 1959: 65, fig. 4; Radoman, 1963a: 79, fig. 8; 1963b: 96, fig. 8; 1983: 119-120, table 7, pl. 8, fig. 140). Dimensions: height = 1.22-1.47 mm; diameter = 1.05-1.18 mm (Radoman, 1983: 206, table 7). Operculum thin, paucispiral, without out- growth at centre of inner side (Hadzisce, 1959: 65; Radoman, 1983: 119). Body unpigmented; eye spots absent (Radoman, 1983: 120). Male genitalia with penis long, cylindrical, slightly dilated at about 2/3 of penis length, then tapering to end in rather pointed tip; slightly evident lobe on left side, level with subapically dilated portion (Radoman’s (1983: 119) diagnosis reports: “. . . without clear outgrowth”) (Hadzisce, 1959: 65, fig. 4; Rado- man, 1963a: fig. 16; 1963b: fig. 16; 1983: 119, fig. 66). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle small but proportionally rather developed; bursa copulatrix very small, with long, slender duct entering bursa on anterior side; seminal groove running along ventral side of capsule gland (Radoman, 1963a: 79, fig. 16; 1963b: 96, fig. 16; 1973a: 6; 1983: 40, 114, 119, fig. 66). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long, robust central denti- cle and about 4 smaller denticles on both sides in decreasing order of size; one basal cusp at point where each lateral wing arises from face of central tooth; lateral teeth rake- like, apically enlarged, their anterior margin with about 10 denticles, central of which longer and larger; first marginal teeth rake- shaped, with long lateral wing and elongated cutting edge with long row of about 20 small denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cut- ting edge carrying rather long row of very small denticles (Hadzisce, 1959: 65, fig. 5). Stomach without posterior caecum; intes- tine unknown (Radoman, 1973a: 6; 1983: 40). Osphradium and ctenidium unknown. Nervous system with long pleuro-supraoe- sophageal and slightly shorter pleuro-suboe- sophageal connectives (Radoman, 1963a: fig. 18b; 1963b: fig. 18b; 1973a: 6; 1983: 119). Taxonomy Radoman (1973a: 8) established Zaumia without a description or definition, but only by combining it with two available species group names. After 1930, this condition does not make a generic name available (ICZN, 1999: Art. 13.6). Zaumia is here considered a dis- tinct genus, but its relationships to Pseudoho- ratia Radoman, 1967, need to be clarified. Za- umia is characterized by: shell very small, valvatiform, conical-ovoid or depressed; oper- culum without peg; penis with one simple lobe; female genitalia with proximal seminal receptacle and very small bursa copulatrix, with long anterior duct; central tooth with one pair of basal cusps. Radoman (1983) included another species from Lake Ohrid in Zaumia: Z. sanctizaumi (Radoman, 1964), distinct from Z. kusceri by valvatiform-planispiral shell. STATUS AND RELATIONSHIPS OF HAUFFENIA The western Palaearctic hydrobiids have rather constant anatomical organization, ex- cept for some features of the penis in males and the renal oviduct in females (Tables 1, 2, Fig. 180). Taxonomy at the rank of genus (and sometimes also at that of family) has thus been traditionally based on penis structure and the number and position of the sac-like structures associated with the renal oviduct (bursa copulatrix and seminal receptacle/s). The penis offers such characters as shape, presence of one of more lobes and their shape, glandular structures in the penis or lobes, papilla or filament at penis tip, stylet- like structures at penial duct opening, and po- sition of penial duct inside penis. These char- acters are generally constant in groups of species and are therefore good diagnostic features at the rank of genus. However, cases are known in which some of these characters are absent in one species of a genus (e.g., Pezzolia; Fig. 180). In supposedly related groups of species, the number and position of the sac-like struc- tures associated with the renal oviduct, bursa copulatrix, and seminal receptacles (one or two) are considered more important (Davis 8 Carney, 1973). Also in this case, however, there are some exceptions, for example, the bursa copulatrix may be reduced, present or 158 BODON, MANGANELLI & GIUSTI MALE Penis with stylet Penis without stylet Penis with/ Penis with Penis with without lobe/s simple lobe/s | glandular lobe/s FEMALE Seminal Bursa receptacle/s copulatrix 4 р | | Normal BC Г Bracenica | or with long duct Daphniola | Gocea | РУК + DSR Horatia Karevia Fissuria Ohridohauffenia Ohrigocea у Prespolitorea Pseudoislamia | Reduced BC | and duct | Pezzolia p.p. MW | + | BC absent Islamia p.p. Islamia p.p. Pezzolia p.p. al mem AE Normal BC with anterior duct Kerkia Only DSR 7 ND BC with posteroventral or JNO 0) lateroposterior duct BC smaller than У 2 0 po Only PSR > Normal BC or with long duct ub iR —+ Lyhnidia Pseudohoratia Strugia Zaumia Reduced BC and duct 3 Hauffenia + Penis lacking lobe/s Sardohoratia Pezzolia p.p. Pezzolia p.p. Arganiella Hadziella Heraultia Dabriana FIG. 180. Synopsis of the European hydrobiid genera with valvatiform shell arranged according to the main anatomical characters of the genitalia (Dalmatella not included because anatomy unknown; it is identified by its keeled shell). The presence of the distal seminal receptacle needs to be confirmed in the type species of Hadziella (the genus is identified by its planispiral shell with reflected peristome). For each genus, only the characters of the type species or closely related species have been considered. Acronyms: BC bursa copulatrix, DSR distal seminal receptacle (first seminal receptacle), PSR proximal sem- inal receptacle (second seminal receptacle). REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 159 TABLE 1. List of the main characters used in discussion of the status and relationships of Hauffenia. The character-state “O” was assigned to Hydrobia, because this genus is considered to be one of the primitive extant hydrobiids (Ponder, 1988a). Protoconch Teleoconch lal T2 Surface depressions: absent (0); pits (1). Spirals: absent (0); spiral threads (1); spiral grooves (2). Wrinkles: absent (0); wrinkles or malleations (1). Shape: ovate-conic (0); planispiral (1); valvatiform (2); trochiform (3); neritiform (4); ovate (5); conic (6); elongate-conic (7). Spiralization: spire entirely coiled (0); despiralized only near aperture (1); almost entirely despiralized (2); horn-like, uncoiled (3). Whorl translation: apex of shell prominent (0); apex of shell flat (1). Whorl outline: flat (0); convex (1); shouldered (2). Color: absent (0); present (1). Microsculpture — Surface depressions: pits absent (0); pits present (1). Spiral microsculpture: absent (0); threads (1); grooves (2). Spiral sculpture: absent (0); crests (1); keel(s) (2). Axial sculpture: growth lines only (0); rounded ribs (1); lamelliform ribs (2). Spines: absent (0); present (1). Aperture: peristome Al A2 A3 Umbilicus U1 Operculum O1 O4 O5 Head-foot H1 Pallial cavity C1 C2 C3 C4 Condition of outer lip relative to rest of apertural plan: simple (0); reflected (1); with varix at edge of outer lip (2); with varix behind outer lip (3). Shape of adapical and abapical portions of outer lip: simple (0); adapically sinuous (1); abapically sinuous (2); adapically and abapically sinuous (3). Inclination of outer lip relative to coil axis: orthocline (0); prosocline (1); opisthocline (2). Size: absent (0); narrow (1); wide (2). Overall shape: ovate (nucleus central or subcentral) (0); elongate-ellipsoid (nucleus sub- marginal) (1); circular (nucleus central) (2). Coiling: paucispiral (less than four whorls) (0); multispiral (four or more whorls) (1). Nuclear thickening and peg: nucleus simple (0); nucleus area thickened (1); nucleus raised into arched peg (2); nucleus raised into spiral peg (3). Crest on inner surface: inner surface simple (0); inner surface with crest (1). Frill: absent (0); slightly developed (1); highly developed (2); forming solid ridge (3). Eyes: present, normal size (0); reduced (1); absent (2). Metapodial tentacle: absent (0); single tentacle present (1). Posterior pallial tentacle: present (0); absent (1). Ctenidium: present (0); absent (1). Osphradium — size: more than 66% of pallial cavity length (0); 50-66% of pallial cavity length (1); less than 50% of pallial cavity length (2). Osphradium — position relative to ctenidium: opposite posterior part of ctenidium (0); opposite middle of ctenidium (1); opposite anterior part of ctenidium (2); inapplicable (ctenidium absent) (-). Osphradium — overall shape: more than three times longer than broad (0); two to three times longer than broad (1); less than twice as long as broad (2). Central teeth — shape:trapezoidal (0); square (1); triangular (2); broadly rectangular (3). Central teeth — excavation of base: less than 25% of tooth height (0); 25%-50% of tooth height (1); more than 50% of tooth height (2). Central teeth —basal tongue length: absent (0); shorter than lateral wing (1); about equal to lateral wing (2); longer than lateral wing (3). Central teeth — basal tongue shape: narrow V-shaped (0); broad V-shaped (1); U-shaped to square (2); slightly convex (3). Central teeth — number of pairs of basal cusps: one pair (0); two pairs (1); three or more pairs (2); basal cusp absent (3). Central teeth — position of basal cusps: all cusps arise from lateral wing (0); one or more cusps arise from tooth face (1); inapplicable (basal cusps absent) (-). Central teeth — relative size of basal cusps: inner cusp larger (0); all cusps equal-size (1); inapplicable (one or no cusps) (-). 160 BODON, MANGANELLI & GIUSTI TABLE 1. (Continued) R8 RQ R10 R11 R12 Stomach Sil S2 Intestine 11 12 Lateral teeths — overall shape: outer margin straight or almost (0); outer margin with dis- tinct concave bend (1). Lateral teeth —shape of face: taller than wide (0); square (1). Lateral teeth —basal projection: absent (0); present (1). Lateral teeth —length of cutting edge relative to outer wings: cutting edge much shorter (0); cutting edge slightly shorter to about equal in length (1). Inner marginal teeth —cusp size: cusp larger than those of outer marginal teeth (0); cusp about as large as those of outer marginal teeth (1). Posterior caecum: present (0); rudimentary or absent (1). Shield caecum: absent (0); present (1). Coiling: simple coil around style sac (0); coil around style sac with additional coil on dorsal side of style sac (1); simple coil far from style sac (2). Shape of rectum within pallial cavity: straight or with slight bend (0); with U-shaped bend (1); with S-shaped bend initiated to left (2); with S-shaped bend initiated to right (3). Female reproductive system F9 F10 F11 F12 F13 F14 wale F16 F17 F18 F19 F20 Pigmentation of coiled oviduct: absent (0); present (1). Overall coiling pattern: single bend or loop (0); two or more bends or loops (1); no loop (2). Type A sperm duct: absent (0); present (1). Number and position of seminal receptacles: one distal (RS1) (0); one proximal (RS2) (1); two seminal receptacles (one proximal, one distal) (2); none (3). Relative size of seminal receptacles: distal (RS1) a little larger or equal to proximal (RS2) (0); distal (RS1) much larger than proximal (RS2) (1); distal (RS1) smaller than proxi- mal (RS2) (2); inapplicable (one or no seminal receptacle) (-). Position of proximal seminal receptacle (RS2) relative to end of loop: at end of loop (0); close to end of loop (1); inapplicable (proximal seminal receptacle absent) (-). Position of proximal seminal receptacle (RS2) relative to bursa copulatrix: all or mostly anterior to bursa (0); lying against bursa (1); behind bursa (2); inapplicable (proximal receptacle or bursa absent) (-). Shape of proximal (RS2) seminal receptacle: elongate (0); pyriform to globular (1); inap- plicable (proximal receptacle absent) (-). Duct of proximal (RS2) seminal receptacle: no distinct duct (0); distinct duct shorter than seminal receptacle (1); duct much longer than seminal receptacle (2); inapplicable (proximal receptacle absent) (-). Position of distal seminal receptacle (RS1) relative to end of loop: far from end of loop, near bursa copulatrix duct (0); closer to end of loop than to bursa copulatrix duct (1); inapplicable (distal seminal receptacle or bursa absent) (—). Position of distal seminal receptacle (RS1) relative to bursa copulatrix: all or most anterior to bursa (0); lying against bursa (1); behind bursa (2); inapplicable (distal receptacle or bursa absent) (-). Shape of distal (RS1) receptacle: elongate (0); pyriform to globular (1); inapplicable (distal receptacle absent) (-). Duct of distal (RS1) seminal receptacle: no distinct duct (0); distinct duct shorter than seminal receptacle (1); duct much longer than seminal receptacle (2); inapplicable (dis- tal receptacle absent) (-). Dimension of posterior bursa copulatrix relative to albumen gland: medium to large (0); rather small (1); very small or rudimentary (2); inapplicable (no bursa) (-). Bursal duct: anteroventral (0); anterodorsal (1); anterior (2); posterodorsal (3); pos- teroventral or posterior (4); posterolateral (5); inapplicable (no bursa) (-). Opening: bursal duct opens into coiled oviduct (0); bursal duct opens into spermathecal duct (1); bursal duct opens into cordoseminal duct (2); inapplicable (no bursa) (—). Length of bursal duct: about 50-100% of bursa length (0); less than 50% of bursa length (1); longer than bursa (2); inapplicable (no bursa) (-). Glandular zones: homogeneous capsule gland (0); two or more glandular zones (1). Spermathecal duct: absent (0); common opening in capsule gland in anterior pallial cavity (1); opens separately to anterior pallial cavity (2); opens separately about halfway along capsule gland (3); opens separately to posterior pallial cavity (4). Ending of uteral gland: not far from pallial margin (0); far from pallial margin (1). Male reproductive system M1 M2 Overall shape of penis: gradually tapering (0); broadly triangular (1); rectangular (2); strap-like, i.e., flat (3). Shape of distal end: tapered (0); blunt (1); expanded (2); subapically expanded and tapered at tip (3). REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 161 TABLE 1. (Continued) M3 Distal papilla: absent (0); present (1). M4 Corneous stylet: absent (0); present, at tip (1); present, inside apex (2). M5 Pigmentation of penis: absent (0); present (1). M6 Simple lobe/s (non-glandular or glandular) on right or upper edge: absent (0); apical (1); lateral, in distal half (2); lateral, in proximal half (3). M7 Simple lobe/s (non-glandular or glandular) on left edge: absent (0); apical (1); lateral, in distal half (2); lateral, in proximal half (3). M8 Non glandular pleat on lower edge: absent (0), present (1). M9 Glandular area inside simple lobe/s: absent (0), apocrine glands (1); internal glandular fields (2); inapplicable (no lobe) (>). M10 External glandular fields or ridges: absent (0); present (1). M11 Sucker-like apocrine glands: absent (0); present (1). M12 Tubular penial gland —number: none (0); one (1); two or more (2). M13 Tubular penial gland — position: absent (—); in penis only (1); in penis and haemocoel (2). M14 Penial duct — position: near centre (0); near right edge (1). absent in a genus (and also in a species, see the case of Pezzolia). Clear relationships between Hauffenia, Lobaunia, Neohoratia, Vrania and Erythropo- matiana emerge from the redescriptions of their type species. In fact, they share the fol- lowing combination of characters: e male genitalia with penis apex rounded; pe- nial duct subcentral; terminal portion of pe- nial duct (immediately before opening) with very small stylet; mass of refringent cells in- side right side of penis apex; variable num- ber (0-3) of more or less evident penial lobes on left side of penis near apex; + female genitalia with bursa copulatrix and proximal seminal receptacle arising from distal renal oviduct; bursa copulatrix very small, with very short duct; proximal semi- nal receptacle shorter than bursa copula- trix. This combination is unique and enables easy taxonomic identification of the Hauffenia group among all anatomically known valvati- form and non-valvatiform genera from all over the world (for the status and identification of all European valvatiform hydrobiid genera, see Tables 3, 4; Fig. 181). Only two characters are available for the subdivision of Hauffenia: the opercular peg and the penial lobes. Hauffenia has an evi- dent peg at the centre of the inner face of the operculum (very developed in H. tovunica Radoman, 1978, rather developed in H. tellinil), Vrania has a smaller peg, Neohoratia and Erythropomatiana lack a peg. As a first conclusion, Erythropomatiana can be pro- posed as a junior synonym of Neohoratia. Oddly, Radoman never considered or dis- cussed Neohoratia when introducing (Rado- man, 1978) and later redescribing (Radoman, 1983) Erythropomatiana. Radoman (1978, 1983) also included a species, E. verdica Radoman, 1978, which is a junior synonym of N. subpiscinalis, the type species of Neoho- ratia in Erythropomatiana (see Taxonomy in Hauffenia subpiscinalis). Vrania was introduced as a subgenus of Hauffenia by Radoman (1978), on the basis of a single diagnostic character: the smaller op- ercular peg. Study of peg variations demon- strates that in specimens of H. tellinii from the type locality, the peg is more dilated apically and never as reduced as in V. wagneri. How- ever, its dimensions vary over a continuum (Figs. 69-71) from that of H. tellinii to that of V. wagneri. Hence, the peg is not a character sufficient to support differentiation of Vrania with respect to Hauffenia at the rank of genus or subgenus. Together with other possible characters, it could at most support differenti- ation at species or subspecies level. In con- clusion, we fully agree with Haase (1992) in considering Vrania a junior synonym of Hauf- fenia. The remarkable concordance of the genital and other anatomical characters in Hauffenia tellinii and Neohoratia subpiscinalis and the variation of the opercular peg from very well developed in H. tovunica Radoman, 1978, to rather well developed in H. tellinii, and small in V. wagneri, throw doubts on the value of the character “peg absent” in N. subpiscinalis and E. erythropomatia, and consequently on the validity of Neohoratia (with Erythropomatiana as a junior Synonym) as a generic taxon dis- tinct from Hauffenia (with Vrania as a junior synonym). Apart the absence of an opercular peg, N. 16 № TABLE 2. Character states (see Table 1) in the European hydrobiid genera with valvatiform shell and in a selection of the best known European hydrobiid genera without valvatiform shell. The table also includes four taxa proposed as junior synonyms of Hauffenia (Erythropomatiana, Neohoratia, Vrania, and Lobaunia). For each genus, only the characters of the type species have been considered or those of the most closely related species, if the anatomy of the type species is not known. ? = no data. CHARACTERS T2 ТЗ T4 T5 T6 T7 T8 T9 T10 A1 A2 АЗ U1 O1 02 O3 O4 O5 Н1 H2 C1 C2 C3 C4 C5 Ri R2 R3 R4 RS R6 R7 RB RI R10R11R12 Panero P1 TAXA 1 it 1 212000 020 4 0 00 0 0 0 0 0 1 4 1 0 0 0 0 2 0 0 Arganiella оо 2 0 22 2 3 ? Bracenica Dabriana 1 1 % 2 0 0° 2 0050, 0 2 0 оо ? 0 00 0 0 0 0 0 ? 2 0 0 0 Dalmatella 2 оо ? Я 4 4 1 12272 CO OO 07 0-0 22 10 OOO 1 4 1 1 1 зоо Daphniola 2 2 12 0 0 00 0 0 0 0 0 0 0 2 0 0 00 2 2 0 2 0 0 0 0 0 0 0 2 ‘Erythropomatiana’ Fissuria Gocea 1 0 0 0 Oo 0 0 0 Oo 0 22000020 4 0 BODON, MANGANELLI & GIUSTI 3 0 0 01 AO) оо оо оо O 01 0 12 0 0 % ODE PEN ? Ор E20 2 4 4 2 0 2 0 2 0772 0 0472 62 2 2 CPE? OF 22 ? 1 1 1 1 1 2 4 ? ? 4 2 2 1 ? 1 ? 2 2 2 2 2 7 2 2 2 2 2 2 2 2 2 2 ? 1 01 0 0 02 ? 0 0 ? 0 ? ? 1 ? 0 ? 4 1 1 1 4 1 1 1 1 1 1 1 1 1 1 1 1 OO О 2° 2 0.3 0 0 2 0 2722097070720 20 2 4 OO 3 о OO 0710" 0 2200 0 0 0 0 0 0 0 0 707707707 0 ооо 2 22 0) 22 207 20 O &0 1 1 2 201001000 2121000020 te 22 OOO 501000 22 Om 10 0101250 fo 2 2 2 2272, 0) 9 0' 0 FO 2 0 2 ee Oe OOS AO 0240 2200 D SOIR DONC AO 1 22000020 2 2 4 2 2 2 1 1 4 4 4 4 1 4 1 1 1 1 1 1 1 1 1 ? 0 00 0 0 0 0 0 0 00 0 0 0 0 0 0 000 0 0 0 0 0 0 00 0 0 0 0 0 2 0 02 00 0 0 0 1 0 0 0 0 0 0 0 0 4 0 000 0 0 0 2 0 1 0 00 0 0 0 0 0 2750710) 2700" ON OO 2 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0.07100" 207.079 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 00 0 0 0 0 1 1 1 1 1 1 1 4 1 1 1 1 1 4 20 02 IA E 270070 0 0 1 12 0 0 1 1 2 0 0 © 3 0-0 ? 1 зоо 00200 ? ? 2 0 0 1 2 0 0 ? 2 0 0 1,300 0 220 0 00300 1 0 0 0 2 2 0 ? 2 0 2 0 2 1 0 0 0 0 4 ? 4 1 ? 1 ? ? 0 в Ohridohauffenia Ohrigocea Pezzolia Pseudoislamia Haaziella Hauffenia Heraultia Horatia Islamia Karevia Kerkia ‘Lobaunia" Lyhnidia ‘Neohoratia” Prespolitorea Pseudohoratia Sardohoratia Strugia ? ? ? ? 2 4 4 1 1 1 4 1 4 1 1 1 4 4 4 1 4 4 4 4 2 090520" 700 2-10 Zee OS NODO 72,530 2 4 01 0 o 0 0 OM ON OM 0 0 1 3 0 0 0 0 ‘Vrania” Zaumia 4 0212 ООО 2540 4 4 1 4 4 4 1 1 4 4 050 0 207 50.707070 o 0 0 0 ON 0 0 0 ооо ооо ооо ооо ооо 1 4 4 4 4 012 0 0 "0 0 0 0 0 0 0 0 0 0 0 Adriohydrobia Alzoniella Avenionia JAN 2 00202207208 27:0 1 0707070707070 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 0 0 0 3 0 0 0 0 0 0 0 | ON АН 0272727070 1 2 2 2 1 1 1 1 1 1 4 0 0 4 4 Belgrandia 0 ? оо 1 4 1 Belgrandiella Bythinella 2 1 022 2 0 9 0 00 0 0 0 0 2 0 000 0 0 0 0 0 0, 100: 10. 0» 0 1 6 00 4 0 1 1 1 1 1 1 4 1 1 0 1 OF 0 ORIG? 202505507 707 70720 2055407 2020 Bythiospeum £ 2 0’ 305507 707.0 00 120220730 0% 40 0 4 Clameia 1 2-22 1 1 0 0 Graecoanatolica Graziana 01 0 1 1 ПО ООО 10250 209270220 0 0 TABLE 2. (Continued) CHARACTERS A2 АЗ U1 O1 O2 O3 O4 O5 H1 H2 C1 C2 C3 C4 C5 Ri R2 АЗ R4 А5 Аб R7 RB R9 А10В11 В12 PEPA STATS Gs И 18s Toe OTAN P1 TAXA 0.22200 1 ооо 4 OOO 021022102707 о 0 4 1 ооо ооо 305 AO OA 02207202 (00 00 60 0) о 1 1 0 Heleobia 4 0-0 1 1 4 4 Hydrobia REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS + сое SS or os | OO Te © @rorerrrrorrerrrrrro = р NE NN | N с ее ое (Ne SiS еее соо-тооосоос<оосоосоо-тососое Е оооооосо - a — a 2 Sroarrzoo0oN0o000n0508 5 9900-0000 -oraunarraerorannrran 2 [|o|«ooro rrrauaauae-aanaaaurauanaae 20000000 CHNNANNNNNENNNNNr-ANNUT—- SE | ONOSANaAa "oo0o00000-o000000000r 2|ooo«"o0o8o -raüeroorrrüärrüneorrrn 2 ooo oooo PE or er" = SONO ere хх а < а < < < ме мамам 000-0000 -orooooo00000ou«-oo5o00o 3moo«ormo rer Or s eo ee oo © o o0o000000000000000000 A 00060000 a a о annnco§$oononnnonnooNon г соосоооо со o0000000000000000000 Г со-сооосо N o0o000000000000000000 „|oorworuaN © ONDODODOOATODODOoooco. г ооосооосоо LO 2c00000000000000000000 [| AAA NA + D0000000000000000000 „|oon"onrOo oO Weare Cite fe ao ee ee Ae Orrco|rr u © esoror-rrToranosrooss-ore г Orrtr|rr PTrrrroororrrrrrooo0or 7|oou«-o|oo o ANS O OO SOOO | DEZE | oo SISI O O SO SO 5s SOS | |э|сеобо eolleooooqooocoo-oooo El === 29 Sooo oo ооо ооо 0000068 ooo || CIA ASIS oroooocororororooocoo fl |0 | соооо eoooooo-cooooooooooo || | = | са os eo0000000000000r-o000 {| омося-ямчм rrrrrrorrrrrrrrorr"rrr О ооосоооо oo6o00o0-o0o000060 000000800 | SS «osos 919 9 9/9 соо SOS Soo SSSos Tl +... ses коохчооюкоюхюююноююоюо чм мессесемаес SCA (©) NO CS ar Or Er CET OO, MOROS ooo госоочо-ссоооос тосе 9oocoo«-o0o000 (POMS KS) LSS) KS) TI IE 975 (еее on Narr SS Zr Er gr E a 2 w < a a = Au = He Q Ye 5395 = E OCS ey au = AZ US 5 а осо SOS on, 8a 8 ЗЕ Rs сео ася оо ба асе Oa et Ses ‚I DBECLLOS ооо Е Е Зоо ERTTEEER SONO nes E ES Si cos es 50 SEESES SSR TERS ESSESES PERES = я = = = ENS AL DOY Sas SS 0/12 Gone a 00 El a ag O 0100 ооо 0770770 OO 10) 10) OOF 0) 0) 707707207 27207 07.07.0770 ORO OO OP ооо ооо ооо 1 0 00202 00 Оооо 4 Hadziella 0’ 2 0 О 02 0 4 1 0 4 Hauffenia Heraultia Horatia OF OF OM 0) 205 © OS 1 4 4 О о li OF MORO OO 0.0 + 2050 0 оо 01 4 1 4 07 207207 2 0) OF 10/00 D 002222. 072.052005202.052.02202205. ооо 000 4 4 0 0 Islamia о 1 OF 40) 08 07 0 0 1 Karevia Kerkia OOOO OLAS M OMOMOMO 2 OOO OO 4 252400 ON 0%2-0 000 0 200 163 912 0) 40; 0,0 4 0 4 0 4 “Lobaunia” TABLE 2. (Continued) 164 CHARACTERS F2 F3 F4 F5 F6 F7 F8 F9 F10F11 F12F13F14 F15 F16 F17 F18 F19 Е20 M1 M2 МЗ M4 M5 M6 M7 M8 M9 M10M11M12M13M14 12 Fi S2 11 S1 TAXA 2 020 0 00 0 0 0 0 0 0.0 2 0 0 0 0 0 2 0 0 2 0 0 0 0 Lyhnidia “Neohoratia" ? ? 0,50, 0 0 0-0 0 0:00. 0.2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 6 0 0 4 1 0 O0 0 0 0 0 0 0 0 0 0 1 1 0 1 4 1 ооо 2 Ohridohauffenia Ohrigocea Pezzolia 0002 2 0 1 ? 0 0 2 000 2 0 0 2 0 0 ооо 0003 3 00000 0 0220 1 1 1 1 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 — 0 Prespolitorea 272790277090 MONO MODO OM 0" ооо ооо о 0 0 0 0 0 0 0 07207 2750 10 “0 2 0 0 1 Pseudohoratia ? Оооо 10270" 109-0) <0 — 1 22020003 OF 27710 30570270) <0" 0.0.0) 700.0: 00 | 1 0 0 0 0 0 0 Pseudoislamia Sardohoratia Strugia 0 00 1 4 fe 2707727207. 0 0.0... 07.0) 07 10) 10) O <0) 0) 00 2 2 0 ? 0 0 2 0 0 0 оо 02070752 то оборо ооо 4 Vrania" BODON, MANGANELLI & GIUSTI Srrrorrrearrroroor и EeOo0000r00000000000 SODOSO90990009- 000000 S09000000606600006008 о тео ег [а SOr9069000000000000 NOOTANODOONANDOODOOO Tr O0OMMNOOOOOOOOO0O0000O Oroor«oooorrrooooo oOo0o000000000090rr000 0.0 0.0.0.0 0.0.0.0 0/0 0000.00 оохохо-осоосоя--оое ооочесоосоосоочоо--о O00000000r-000000000 ooooooroocooox+oooooo O00000000000r-000000o Noo-ocooo | ono ooo oooocoooroo|oroocooocooo NNTONTOY+=<|qnorsrasrosS NTOoDoocooco|orroocooro |o-oorornworr|r{rrr |eoroororrooolrorr jeoeoororr|rro|rrrr |[ococooceos | сое |osoo olor-|lllllllellle ое Ее ~lere lili ЕЕ а ЕЕ опере ето esa О gh Ae ale cline TONNNOODOOOOOOT-ODOAM 000000” 0000000000 Eoroo00000r00ro00000 orooo0000000r00000 “ONDODONO-DOONOA AO ооо о 0.0 0/0/90 SO 1 ео SOOO SOOO OOH OO 0/0/09: O10 rOorrrrrr rrr rorrrre $ ee oe S ваз ооо ева ash? ЕЕ Ч << об0соет55345 0 0 0,70 207207 190720720 ооо о 1 0 70707 0 07 0 Marstoniopsis Mercuria 07270070 1 0 07070771 1 0 0 0 0 0 0 0 4 4 4 1 1 1 1 OOO 0020) 720075020730 4 ооо 0720702020722 ооо ооозоо 207 707 72720707 7020750 0 20 000 3 Moitessieria 0 4 0 1 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0052 0 0 07 0 Orientalina ооо 0.2070 ооо OL 3:10) 07 05-05 50° о OOO 075020520770 2 1 0 4 0 0 0 0 Oo 0 0 4 0 0 0 0 0 0 3 0 0 0 0 0 0 0 4 0 0 0 0 1 Palacanthiliopsis Paladilhia 4 0 0 00 0 0 0 0 0° 0° 07.00 0 2 0 070707007020 Paladilhiopsis Pauluccinella Phreatica 1 ооо ооо ооо 0 4 0 0 0 00.0 0 0 O 0 © OF 50779 OA OM 0:40 707 207 07 0 0 20207507 0 о 1 4 4 4 Plagigeyeria ONO MOTO 0 20" 50 ТОРО 0 0 10" OP 07 30: 207707790207 — 1 Potamopyrgus 07071 ONO 0 OL O08 ONE ON OMS RD 27 ,052:02505 ооо о 1 Pseudamnicola 07-207 0 050350 4 1 0 1 00520 010750 00 000 020 4 1 1 Pseudavenionia Sadleriana 07.0070 IRON 1 1 ооо 9 O52 OOOO: Sardopaladilhia REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 165 TABLE 3. Valvatiform hydrobiid taxa of the genus group from Europe and the Middle East (* taxa not dis- cussed in the paper) and their taxonomic status according to the present revison or the recent literature. Nominal taxon Taxonomic status Adriolitorea Radoman, 1973b, as /slamia (Adriolitorea) Arganiella Giusti & Pezzoli, 1980 Bracenica Radoman, 1973a Dabriana Radoman, 1974 Dalmatella Velkovrh, 1970 Daphniola Radoman, 1973a Daudebardiella* Boettger, 1905 Dolapia Radoman, 1983 Erythropomatiana Radoman, 1978 Fissuria Boeters, 1981 Gocea HadZisce, 1956 Haaziella Kuscer, 1932 Hauffenia Pollonera, 1898, as Horatia (Hauffenia) Heraultia n. gen. Horatia Bourguignat, 1887 Islamia Radoman, 1973a Karevia Radoman, 1973a (not Hadzisce, 1959) Karevia HadZiSce, 1959, as Ohrigocea (Karevia) Kerkia Radoman, 1978 Lobaunia Haase, 1993 Lyhnidia HadZisce, 1959 Mienisiella Schutt, 1991 Naumia Radoman, 1973a Neohoratia Schütt, 1961a, as Horatia (Neohoratia) Ohridohauffenia HadZisce, 1959, as Ohridohoratia (Ohridohauffenia) Ohridosturanya Radoman, 1973a Ohrigocea Hadzisce, 1959, as Ohrigocea (Ohrigocea) Pezzolia Bodon & Giusti, 1986 Prespolitorea Radoman, 1983 Pseudohoratia Radoman, 1967 Pseudoislamia Radoman, 1979 Rotondia Radoman, 1964, as Pseudamnicola (Rotondia) Sardohoratia Manganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998 Sheitanok* Schütt & Sesen, 1991 Strugia Radoman, 1973a Vrania Radoman, 1978, as Hauffenia (Vrania) Zaumia Radoman, 1983 subpiscinalis and E. erythropomatia share an- other character state, the evident penial lobes, in the form of small pimples. Vrania wagneri apart from having a reduced opercu- lar peg, is characterized by one evident wide penial lobe. Hauffenia tellinii, apart from hav- ing a rather developed opercular peg, is char- acterized by wide, but not evident penial lobes, or none at all. Three groups, based on two diagnostic characters, can therefore be identified. The only anatomical difference between the four taxa consists in the different number and junior synonym of /s/amia Arganiella Bracenica Dabriana Dalmatella Daphniola Daudebardiella* junior synonym of Karevia junior synonym of Hauffenia Fissuria Gocea Haaziella Hauffenia Heraultia Horatia Islamia junior synonym of Ohrigocea Karevia Kerkia junior synonym of Hauffenia Lyhnidia junior synonym of /slamia name not available (see remarks to Ohridohauffenia) junior synonym of Hauffenia Ohridohauffenia name not available (see remarks to Ohrigocea) Ohrigocea Pezzolia Prespolitorea Pseudohoratia Pseudoislamia name not available (see remarks to Ohridohauffenia) Sardohoratia Sheitanok Strugia junior synonym of Hauffenia Zaumia evidence of the penial lobes: 2-3, small, evi- dent and roundish (Neohoratia, Erythropoma- tiana); 0-2, large but not evident (Hauffenia); one, evident and wide (Vrania). These differ- ences are not sufficient for a distinction at the rank of genus or subgenus, in view of the marked variability in some of the type species studied by us (see above description and Figs. 22, 73-75, 84, 130, 138, 140, 169). We therefore propose Neohoratia, Vrania, Eryth- ropomatiana as junior synonyms of Hauffenia. Finally, with regard to Lobaunia, Haase’s (1992) interpretation of its female genitalia TABLE 4. Analytical key for the identification of the European valvatiform hydrobiid genera (Dalmatella is not included, because its anatomy is unknown; it is easily identified by its shell characters). For characters used, see Fig. 181. 1a—Three sac-like structures on renal oviduct: a bursa copulatrix 2 (BC) and two seminal receptacles: distal (DSR) and proximal (PSR) 1b — Two sac-like structures on renal oviduct: a bursa copulatrix 12 (BC) and a seminal receptacle (DSR or PSR) or two seminal receptacles (DSR and PSR) 2a — ВС normal-sized, with evident duct 3 2b —BC reduced in size, not larger than a seminal receptacle, without Pezzolia (in part) (Liguria, Italy) evident duct; penis without lobe/s or with one or two glandular lobe/s За — Penis with lobe/s 4 3b — Penis without lobe/s; PSR bent outwards Sardohoratia (Sardinia Island, Italy) 4a —Penis with one simple lateral lobe 5 4b —Penis with 2—4 glandular lobes Fissuria (France, Italy, and Greece) 5a — BC large, with duct about as long as BC 6 5b —BC small, with duct about twice as long as BC Pseudoislamia (Greece) ба —DSR slightly larger or smaller than PSR 7 6b —DSR much larger than PSR and about as long as bursa copu- Bracenica (Montenegro) latrix duct; BC pyriform; operculum with peg 7a—PSR distinct 8 7b —PSR relatively indistinct, bent to adhere to wall of oviduct Horatia (Dalmatia and Macedonia) 8a — ВС roundish 9 8b —BC not roundish 10 9a—PSR much larger than DSR Prespolitorea (Prespa basin) 9b —PSR slightly larger than DSR Daphniola (Greece) 10a—BC pyriform 11 10b —BC arched, kidney- or crescent-shaped; shell with two rows of Karevia (Lake Ohrid) raised nail-like projections 11a—PSR larger than DSR Ohrigocea and Ohridohauffenia (Ohrid basin) 11b—PSR about as long as DSR; last whorl of shell despiralized; Gocea (Lake Ohrid) operculum spiralized on outer face to resemble screw 12a — Seminal receptacle (PSR) at end of loop of oviduct 13 12b —No seminai receptacle (PSR) at end of loop of oviduct (only 19 DSR and BC beyond end of loop) 1За —ВС normal-sized with evident duct, if reduced with long duct; 14 penis with simple lobe 13b —BC reduced in size, without evident duct, or absent (in the lat- 17 ter case DSR present) 14a—Penial lobe large 15 14b — Penial lobe small 16 15a—BC medium-sized Lyhnidia (Ohrid basin) 15b —ВС very small Zaumia (Ohrid basin) 16a—BC medium-sized Strugia (Ohrid basin) 16b — ВС very small; operculum with peg Pseudohoratia (Lake Ohrid) 17a—BC slightly larger than PSR; wide penial apex with stylet and Hauffenia (Austria, NE Italy, subcentral penial duct; operculum with or without peg and NW Balkans) 17b—BC absent and DSR not larger than PSR; penial apex without 18 stylet and with lateral penial duct 18a—PSR usually larger than DSR and with evident duct; penis Islamia (France, Switzerland, S Eu- with one lateral or apical lobe, usually glandular rope, Turkey, Israel and Lebanon) 18b —PSR usually smaller than DSR and without evident duct; Pezzolia (in part) (Liguria, Italy) penis without or with one or two glandular lobe/s 19a—BC larger than DSR 20 19b —BC small, smaller than DSR (which is very large); penis without Dabriana (Bosnia-Herzegovina) lobe/s; caudal tentacle; central tooth of radula without basal cusps 20a — Duct of bursa copulatrix entering bursa on anterior side 21 20b — Duct of bursa copulatrix entering bursa on posteroventral or 22 lateroposterior side; penis without lobe/s 21a—Penis with one simple lobe; operculum with crest on inner face Kerkia (Slovenia) 216 — Penis without lobe/s; operculum at most with circular thicken- Arganiella (Central Italy) ing at centre of inner face 22a — Duct of bursa copulatrix entering bursa on posteroventral Hadziella (NE Italy and NW Balkans) side; shell planispiral 226 — Duct of bursa copulatrix entering bursa on lateroposterior side Heraultia (Hérault, France) REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 167 11 Е NAO о PER 19a 20a 4 и Er Lee es Са Po ee I 22a 22b 21a FIG. 181. Characters used in the analytical key for the identification of genera of European valvatiform hy- drobiids (Table 4). The bar at the end of the oviduct means that another structure (bursa copulatrix or semi- nal receptacle) may be present; the double bar that two other structures (bursa copulatrix and seminal re- ceptacle or two seminal receptacles) may be present. 168 BODON, MANGANELLI & GIUSTI needs confirmation. We propose this genus as a junior synonym of Hauffenia. Concluding, the genus Hauffenia (including Erythropomatiana, Neohoratia, Vrania and Lobaunia) is characterized by: shell very small, conical-valvatiform to planispiral; oper- culum with or without peg; penis with stylet and with or without lobes; female genitalia with proximal seminal receptacle and very small bursa copulatrix; central tooth with one or two pairs of basal cusps. We performed a parsimony analysis on a selection of European valvatiform and non- valvatiform hydrobiid genera in order to verify whether our conclusion about the affinities be- tween Hauffenia, Lobaunia, Neohoratia, Vra- nia and Erythropomatiana was supported by synapomorphies. The data matrix consisted of a selection of 44 of the 62 taxa listed in Table 2. Sixteen genera were excluded be- cause they were not well known (more than eight characters unknown: Adriohydrobia, Bracenica, Clameia, Dabriana, Dalmatella, Daphniola, Graecoanatolica, Karevia, Iglica, Ohriohauffenia, Ohrigocea, Palacanthiliopsis, Prespolitorea, Pseudoislamia, Strugia and Zaumia). All characters were assigned equal weight, and multistate characters were treated as nonadditive. Twenty-two of the 79 characters were parsimony uninformative: six were constant (T8, T10, H2, R10, M3, M13) and 16 were autoapomorphies (T2, T6, T9, Од, 05, СЗ; СА, ВТ, Ril, S2,F2, РБ, ЕЮ, F18, M10, M11). Character polarity was de- termined by outgroup comparison, choosing four different genera as outgroup: Hydrobia (subfamily Hydrobiinae), regarded as one of the most primitive extant genera (Ponder, 1988a); Bythinella, regarded as belonging to the family/subfamily Amnicolidae/Amnicoli- nae (Davis et al., 1985; Hershler & Thomp- son, 1988; Hershler & Ponder, 1988a); Heleo- bia, regarded as belonging to the subfamily Cochliopinae (= Littoridininae) (Davis et al., 1983; Hershler & Thompson, 1992; Hershler & Ponder, 1998); Potamopyrgus, regarded as possibly belonging to the distinct subfamily Potamopyrginae (Boeters, 1988; Ponder, 1988b). 1152 most parsimonious hypotheses were generated by our data matrix. All had 332 steps, with a Consistency Index of 0.425 and a Retention Index of 0.576 (Swofford, 1997). The strict consensus of the trees (Fig. 182) showed that 14 monophyletic groups ap- peared in all trees and that one of them was the Hauffenia group. If shell characters (P1- P3, T1-T10, A1-A3, U1) are excluded (most shell characters are considered to be plastic and subject to convergence; Ponder, 1988a), a total of 8208 most parsimonious hypotheses were generated by our data matrix. All had 250 steps, with a Consistency Index of 0.432 and a Retention Index of 0.571 (Swofford, 1997). The strict consensus of these trees (Fig. 183) showed that eight monophyletic groups appeared in all trees and that one of them was the Hauffenia group. In the two sets of trees, the Hauffenia group is always sup- ported by the following synapomorphies: F4, [proximal seminal receptacle], M2, [penis with apex blunt], M4, [corneous stylet inside apex of penis] and M14, [penial duct running through central portion of penis], but only one of these, M4,, is exclusive. Shell characters played a major role in re- solving phylogenetic relationships among the various genera (Fig. 182): when shell charac- ters are omitted, resolution diminishes and some of the monophyletic groups disappear or change. The cladistic analysis clearly produced a “non-result”. Nevertheless, it was useful to show that morphological characters are not sufficient to reconstruct the phylogenesis of the hydrobiids. A combined approach, involv- ing consideration of morphological and ge- netic data, is the only valid alternative. STATUS OF THE SPECIES PRESENTLY ASSIGNED TO HAUFFENIA Many species from different European lo- calities (and even from outside Europe) have been assigned to Hauffenia and Neohoratia (Kuscer, 1932, 1933a, b; Bole, 1961, 1967a, b, 1979, 1993; Schutt, 1961b, 1962, 1980; Binder, 1966; Angelov, 1967; Radoman, 1973a, 1978, 1983; Bernasconi, 1975, 1977, 1984, 1985, 1988; Maassen, 1975, 1978; Boeters, 1973, 1974, 1981, 1988, 1998; Git- tenberger, 1982; Bole & Velkovrh, 1986, 1987; Boeters & Rolan, 1988; Bech, 1990; Jo- vanovic, 1991; Haase, 1992, 1993; Hinz et al., 1984; Rolan 1997a, b). Most of them, studied on the basis of shell characters only, followed in only a few cases by anatomical study, are in need of revision. As far as possible, we checked the status of each of them by study- ing any new material available. Only a few species from the eastern Alps and Balkans actually belong to Hauffenia (for REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 169 Alzoniella Alzoniella | Pseudavenionia Pseudavenionia An 1 ganiella Arganiella а Haaziella Haaziella Sardohoratia Sardohoratia Heraultia Herauitia Kerkia Kerkia Erythropomatiana En Hauffenia feel Clos Lobaunia Lobaunia Neohoratia to tia UCI Pezzolia Pezzolia вета Fissuria Gocea seso Horatia Horatia Lyhnidia ace A Pseudohoratia Pseudohoratia Avenionia Aven ionia Bythinella Belgrandia Marstoniopsis Belgrandiella Islamia Bythiospeum ETA Istriana Litthabitella a Lanzaiopsis Belgrandia Lithogly E que Pauluccinella Moitessieria A ase ue Belgrandiella Sardopaladilhia pl ol Paladilhiopsis Litthabitella Phreatica Lanzaiopsis Heleobía Lithoglyphus Hydrobia Moitessierla Pseudamnicola Paladilhia RHONE Plagigeyeria Graziana Sardopaladilhia Pauluccinella Paladilhiopsis Islamia Phreatica M CURE) Hydrobia Sadleriana . : Pseudamnicola Orientalina _ Potamopyrgus Marstoniopsis Heleobia Bythinella Alzoniella Alzoniella Pseudavenionia Pseudavenionia Arganiella Arganiella Haaziella Haaziella Sardohoratia Sardohoratia Heraultia Heraultia Kerkia Kerkia Erythropomatiana Erythropomatiana Hauffenia Hauffenia Lobaunia Lobaunia Neohoratia Neohoratia Vrania Vrania Pezzolia Pezzolia Fissuria Fissuria Gocea Gocea Horatia Horatia Lyhnidia Lyhnidia Pseudohoratia Pseudohoratia Avenionia Avenionia Bythinella Bythinella Marstoniopsis Marstoniopsis Islamia Islamia Mercuria Mercuria Sadleriana Sadleriana Orientalina Orientalina Belgrandia Belgrandia Pauluccinella Pauluccinella Graziana Graziana Belgrandiella Belgrandiella Bythiospeum Bythiospeum Istriana Istriana Litthabitella Litthabitella Lanzaiopsis Lanzaiopsis Lithoglyphus Lithoglyphus Moitessieria Moitessieria Paladilhia Paladilhia Plagigeyeria Plagigeyeria Sardopaladilhia Sardopaladilhia Paladilhiopsis Paladilhiopsis Phreatica Phreatica Heleobia Heleobia Potamopyrgus Hydrobia Pseudamnicola Pseudamnicola Hydrobia Potamopyrgus FIG. 182. The strict consensus trees of 1152 most parsimonious cladograms generated by the data matrix (Table 2) using Bythinella, Heleobia, Hydrobia and Potamopyrgus as outgroup for determining character po- larity. 170 BODON, MANGANELLI & GIUSTI Alzoniella Alzoniella Fissuria Fissuria Arganiella Arganiella Avenionia Avenionia Islamia Islamia Belgrandia Belgrandia Belgrandiella Belgrandiella Bythiospeum Bythinella Erythropomatiana Bythiospeum Hauffenia Erythropomatiana Lobaunia Hauffenia Neohoratia Lobaunia Vrania Neohoratia Gocea Vrania Horatia Gocea Graziana Horatia Haaziella Graziana Heleobia Haaziella Heraultia Heraultia Hydrobia Hydrobia Mercuria Mercuria Pseudamnicola Pseudamnicola Istriana Istriana Litthabitella Litthabitella Kerkia Kerkia Lanzaiopsis Lanzalopsis Moitessieria Moitessieria Paladilhia Paladilhia Phreatica Phreatica Plagigeyeria Plagigeyeria Sardopaladilhia Sardopaladilhia Lithoglyphus Lithoglyphus Potamopyrgus Potamopyrgus Marstoniopsis Marstoniopsis Lyhnidia Lyhnidia Pseudohoratia Pseudohoratia Orientalina Orientalina Paladilhiopsis Paladilhiopsis Pauluccinella Pauluccinella Pezzolia Pezzolia Pseudavenionia Pseudavenionia Sadleriana Sadleriana Sardohoratia Sardohoratia Bythinella Heleobia Alzoniella Alzoniella Fissuria Fissuria Arganiella Arganiella Avenionia Avenionia Islamia Islamia Belgrandia Belgrandia Belgrandiella Belgrandiella Bythinella Bythinella Bythiospeum Bythiospeum Erythropomatiana Erythropomatiana Hauffenia Hauffenia Lobaunia Lobaunia Neohoratia Neohoratia Vrania Vrania Gocea Gocea Horatia Horatia Graziana Grazlana Haaziella Haaziella Heleobia Heleobia Heraultia Heraultia Istriana Hydrobia Litthabitella Мегсипа Kerkia Pseudamnicola Lanzaiopsis Istriana Moitessieria Litthabitella Paladilhia Kerkia Phreatica Lanzaiopsis Plagigeyeria Moitessieria Sardopaladilhia Paladilhia Lithoglyphus Phreatica Potamopyrgus Plagigeyeria Marstoniopsis Sardopaladilhia Lyhnidia Lyhnidia Pseudohoratia Pseudohoratia Orientalina Orientalina Paladilhiopsis Paladilhiopsis Pauluccinella Pauluccinella Pezzolía Pezzolia Pseudavenionia Pseudavenionia Sadleriana Sadleriana Sardohoratia Sardohoratia Mercuria Marstoniopsis Pseudamnicola Lithoglyphus Hydrobia Potamopyrgus FIG. 183. The strict consensus trees of 8208 most parsimonious cladograms generated by the data matrix (Table 2) when shell characters (P1-P3, T1-T10, A1-A3 and U1) are excluded. Character polarity was deter- mined by outgroup comparison, choosing four different genera as outgroup: Bythinella, Heleobia, Hydrobia and Potamopyrgus. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS WA 0 50 100 150 200 250 300 имы вы 655 I Chilometri FIG. 184. Geographic range of the genus Hauffenia Pollonera, 1898 (only species certainly belonging to the genus are considerea). the distribution of Hauffenia and other Euro- pean valvatiform hydrobiid genera, see Figs. 184-187; for the status and distribution of all European valvatiform hydrobiid species, see Tables 5, 6). In addition to the type species of Hauffenia, H. tellinii (Pollonera, 1898); Eryth- ropomatiana, H. erythropomatia (Hauffen, 1856); Neohoratia, H. subpiscinalis (Kuscer, 1932); and Vrania, H. wagneri (KuSéer, 1928), they are: H. kerschneri (Zimmermann, 1930), H. media Bole, 1961, H. subcarinata Bole & Velkovrh, 1987, H. tovunica Radoman, 1978, and H. wienerwaldensis Haase, 1992. They are redescribed below (for their identification, see Table 7, Fig. 188). The misidentified species will be discussed in four groups on the basis of their geograph- ical distribution (central-eastern European species, French species, Spanish species, and North American species). Central-Eastern European Species. Some nominal species described from the Balkan Peninsula are assigned to Hauffenia and to Neohoratia and listed as valid taxa by Bole & Velkovrh (1986), although all of them were overlooked by Radoman (1973, 1983). Their distribution far from Slovenia and northern Croatia (where species of Hauffenia live) throws serious doubt on their inclusion in Hauffenia. They are: “Hauffenia” edlaueri (Schütt, 1961b), “Hauffenia” jadertina KuScer, 1933a, “Hauffenia” lucidula (Angelov, 1967), “Hauffe- па” plana Bole, 1961, “Hauffenia” raehlei (Schütt, 1980), “Hauffenia” sinjana (Kuscer, 1933a, “Hauffenia” solitaria Bole & Velkovrh, 1986 (nomen nudum), “Neohoratia” epirana (Schütt, 1962) and “Neohoratia” hadei (Git- tenberger, 1982). For these species (except “H.” lucidula, “H.” raehlei and “N.” epirana), 172 BODON, MANGANELLI & GIUSTI FIG. 185. Geographic range of the genus /slamia Radoman, 1973a. The distribution is probably wider in western and eastern Europe and Asia Minor, where little research has been carried out. lack of anatomical data makes their inclusion in Hauffenia entirely speculative. In the case of “H.” lucidula and “N.” epirana, there is too lit- tle anatomical data available to make a posi- tive generic arrangement. On the other hand, there is sufficient anatomical data on “H.” raehlei to allocate it to another genus, namely Fissuria (see Fissuria raehlei in the “Descrip- tions of some taxa misidentified as Hauffenia species”). French Species. Several very small, valvati- form, freshwater prosobranch gastropods from France were described under the generic name Valvata at the dawn of modern malacology: V. minuta Draparnaud, 1805; V. moquiniana Dupuy, 1851; У globulina Paladilhe, 1866; V. exilis Paladilhe, 1867; V. bourguignati Letourneux, 1869; V. micromet- rica Locard, 1889; and V. turgidula Locard, 1889. Germain (1913, 1931) made the first revi- sion, confirming only four species as valid: V. minuta (with V. turgidula as a junior synonym or form), V. globulina (with V. bourguignati and V. micrometrica as junior synonyms), V. mo- guini [sic] and V. exilis. More recently, Binder (1966) regarded V. minuta and V. globulina as belonging to the same, very variable species. On the basis of available conchological and REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 173 50 100 150 200 250 300 Chilometri FIG. 186. Geographic range of the genera Arganiella Giusti 8 Pezzoli, 1980 (A), Bracenica Radoman, 1973a (B), Dabriana Radoman, 1974 (Db), Dalmatella Velkovrh, 1970 (DI), Daphniola Radoman, 1973a (Dp), Haaziella KuScer, 1932 (Ha), Horatia Bourguignat, 1887 (Ho), Kerkia Radoman, 1978 (K), Pezzolia Bodon 8 Giusti, 1986 (Pz), Pseudoislamia Radoman, 1979 (Ps) and endemic genera of Ohrid and Prespa basins (E) (Gocea HadzZisce, 1956, Karevia Hadzisce, 1959, Lyhnidia HadziSée, 1959, Ohridohauffenia Hadzisce 1959, Ohrigocea HadZiSce, 1959, Prespolitorea Radoman, 1983, Pseudohoratia Radoman, 1967, Strugia Radoman, 1973a, Zaumia Radoman, 1983). anatomical characters, he demonstrated that this species was a hydrobiid, which he as- signed to Hauffenia. Bernasconi (1975, 1977, 1984, 1985, 1988) revised the French Hauffenia and concluded that only one species, Hauffenia (Neohoratia) minuta (Draparnaud, 1805), lived in France. He regarded V. globulina and V. moquiniana as junior synonyms of H. minuta, and, follow- ing Boeters (1974), V. exilis as a species of Horatia. He split H. minuta into four sub- species, two of which he established: H. mi- nuta minuta, H. minuta globulina (Paladilhe, 1866), H. minuta spirata Bernasconi, 1985, and H. minuta consolationis Bernasconi, 1985. The assignment of these taxa to Hauf- fenia (Neohoratia) was based on the small bursa copulatrix and single seminal recepta- cle (characters typical of Hauffenia) and on the absence of the opercular peg (a character considered typical of Neohoratia). Unfortunately, Bernasconi’s papers contain many mistakes, including misinterpretation of female genital structure. Our study of the French Hauffenia demonstrated that the two sac-like structures were two seminal recepta- cles, not a bursa copulatrix and a seminal re- ceptacle. This and the structure of the penis (presence of glandular lobe on left side; ab- sence of glandular mass in apical portion to right of penial duct; absence of stylet-like structure at tip) support inclusion of the 174 BODON, MANGANELLI & GIUSTI FIG. 187. Geographic range of the genera Fissuria Boeters, 1981 (F), Heraultia, n. gen. (H) and Sardohora- tia Manganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998 (S). French Hauffenia species in the genus /s- lamia (see “Descriptions of some taxa misidentified as Hauffenia species’). Our study supports placement of V. exilis in a new genus (see Heraultia n. gen. in “De- scription of a new valvatiform genus from France’). Spanish Species. In a recent revision of the western European hydrobiids, Boeters (1988) assigned a number of Iberian valvatiform species to two genera: Horatia and Neohoratia. According to him, Horatia includes two Iber- ian species: H. gatoa Boeters, 1980; and H. (?) sturmi (Rosenhauer, 1856). The latter species was recently revised by Ramos et al. (1992) on topotypical material. None of these taxa belong to Horatia, notwithstanding the fact that they have a wide bursa copulatrix similar to that of species of Horatia (see de- scription of Horatia). In fact, Paludina sturmi has the penis divided into a wide basal portion and a very long, slender tip with a small lobe near its base, and the female genitalia have only two sac-like structures: a large bursa copulatrix and a seminal receptacle (proximal or RS2) (Boeters, 1988: 220-221, figs. 160, 161, 172, 173; Ramos et al., 1992: 485). The same is true of Horatia gatoa, although this REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 175 TABLE 5. Valvatiform hydrobiid taxa of species group from Europe and the Middle East (* taxa not discussed in the paper) and their taxonomic status according to the present revision or the recent literature (when the generic name is in quotation marks, allocation of the taxon is dubious or very uncertain). Nominal taxon albanica, Horatia— Bourguignat, 1887 amidicus*, Sheitanok— Schutt & Sesen, 1991 anatolica*, Islamia— Radoman, 1973a anti, Hadziella— Schutt, 1960 asiana*, Daudebardiella— Boettger, 1905 ateni, Microna— Boeters, 1969 azarum, Neohoratia- Boeters 8 Rolan, 1988 balcanica, Pseudoislamia— Radoman, 1979 bendidis*, Islamia— Reischútz, 1988 birsteini*, Horatia— Starobogatov, 1962 borutzki”, Horatia- Zhadin, 1932 bosniaca*, Islamia— Вадотап, 19734 bosniaca, Dabriana— Radoman, 1974 boui, Fissuria—Boeters, 1981 bourguignati, Valvata—Letourneaux, 1869 brezicensis, Kerkia—Bodon 8 Cianfanelli, 1996 brusinae, Horatia— Radoman, 1953 burnabasa*, Horatia—Schútt, 1964 cianensis, Islamia— Bodon, Manganelli, Sparacio & Giusti, 1995 consolationis, Hauffenia minuta— Bernasconi, 1985 coronadoi, Valvata— Bourguignat, 1870 danubialis, Lobaunia— Haase, 1993 deminuta, Hadziella ephippiostoma— Bole, 1961 depressa, Pseudamnicola— Radoman, 1957 drimica, Pseudamnicola (Rotondia)— Radoman, 1964 edlaueri, Horatia (Hauffenia)— Schütt, 1961b ephippiostoma, Hadziella— KuScer, 1932 epirana, Horatia (Neohoratia)— Schutt, 1962 erythropomatia, Valvata— Hauffen, 1856 exigua, Valvata— Schmidt, 1856 exilis, Valvata— Paladilhe, 1867 fezi, Valvata (? Tropidina)— Altimira, 1960 fontinalis, Horatia— Bourguignat, 1887 gaillardoti, Valvata (Cincinna)— Germain, 1911 gaiteri, Islamia— Bodon, Manganelli, Sparacio & Giusti, 1995 gasulli, Hauffenia (Neohoratia)— Boeters, 1981 gatoa, Horatia—Boeters, 1980 gjorgjevici, Lyhnidia— Hadzisce, 1959 gjorgjevici, Ohridohoratia (Ohridohauffenia)—Hadzisce, 1959 globulina, Valvata—Paladilhe, 1866 globulus, Amnicola— Bofill, 1909 graeca*, Islamia— Radoman, 1973a graeca, Daphniola— Radoman, 1973a hadei, Horatia (Neohoratia)— Gittenberger, 1982 hadzii, Lyhnidia—Hadzisce, 1959 hellenica, Valvata (Cincinna)— Westerlund, 1898 islamioides, Sardohoratia— Manganelli, Bodon, Cianfanelli, Talenti 8 Giusti, 1998 jadertina, Hauffenia-Kuscer, 1933a karamani Lyhnidia— Hadzisce, 1959 karevi, Ohrigocea (Ohrigocea)—HadZiSce, 1959 kerschneri, Horatia erythropomatia— Zimmermann, 1930 klecakiana, Horatia— Bourguignat, 1887 knorri*, Horatia— Schütt, 1961 Taxonomic status junior synonym of Horatia klecakiana Sheitanok amidicus* Islamia anatolica* Haaziella anti Daudebardiella asiana* Islamia ateni Islamia azarum Pseudoislamia balcanica “Islamia” bendidis* “Horatia” birsteini* “Horatia” borutzkii* Islamia bosniaca* Dabriana bosniaca Fissuria boui possible junior synonym of Islamia globulina Kerkia brezicensis Pseudohoratia brusinae Islamia burnabasa* Islamia cianensis Islamia consolationis “Islamia” coronadoi Hauffenia danubialis Hadziella deminuta Ohridohauffenia depressa Ohridohauffenia drimica “Hauffenia” edlaueri Haaziella ephippiostoma “Islamia” epirana Hauffenia erythropomatia Daphniola exigua exigua Heraultia exilis “Islamia” fezi junior synonym of Horatia klecakiana Islamia gaillardoti Islamia gaiteri “Horatia” gasulli “Horatia” gatoa Lyhnidia gjorgjevici junior synonym of Ohridohauffenia depressa Islamia globulina Islamia globulus globulus Islamia graeca* (junior synonym of “/s/amia” epirana?) junior synonym of Daphniola exigua exigua “Hauffenia” hadei Lyhnidia hadzii junior synonym of Daphniola exigua exigua Sardohoratia islamioides “Hauffenia” jadertina Lyhnidia karamani Ohrigocea karevi Hauffenia kerschneri Horatia klecakiana “Horatia” knorri 176 BODON, MANGANELLI & GIUSTI TABLE 5. (Continued) Nominal taxon krkae, Hadziella— Bole, 1992 kusceri, Hauffenia—Bole, 1961 kusceri, Horatia—Hadzisce, 1959 lacustris, Pseudamnicola (Rotondia)— Radoman, 1964 lagari, Pseudamnicola— Altimira, 1960 latina”, Islamia— Radoman, 1973a letourneuxi, Horatia— Bourguignat, 1887 ljovuschkini*, Horatia— Starobogatov, 1962 loichiana, Hauffenia kerschneri— Haase, 1993 lucidula, Horatia (Hauffenia)— Angelov, 1967 lucidulus [sic], Horatia (Hauffenia)— Angelov, 1967 lyhnidica, Pseudamnicola (Ohrigocea)— Radoman, 1963a macedonica, Sadleriana—Kuscer, 1936 malaprespensis, Prespolitorea—Radoman, 1973a media, Hauffenia— Bole, 1961 michleri, Hauffenia—Kuscer, 1932 micrometrica, Valvata—Locard, 1889 mienisi, Mienisiella— Schutt, 1991 miladinovorum, Ohrigocea (Karevia)— Hadzisce, 1959 miljackae*, Dalmatella— Bole & Velkovrh, 1986 minuta, Pseudamnicola— Radoman, 1955 minuta, Valvata— Draparnaud, 1805 moquiniana, Valvata— Dupuy, 1851 naegelei*, Daudebardiella— Boettger, 1905 novoselensis, Horatia— Radoman, 1966 obliqua, Horatia—Bourguignat, 1887 obtusa, Horatia—Bourguignat, 1887 ochridana, Valvata (Atropidina)— Polinski, 1929 ohridana, Gocea— Hadzisce, 1956 ohridana, Strugia— Radoman, 1973a ornata, Pseudamnicola— Radoman, 1957 palustris, Horatia— Bourguignat, 1887 pangaea, Horatia (Daphniola) exigua— Reischútz, 1984 parvulus*, Lithoglyphus—Naegele, 1894 pescei, Arganiella— Giusti & Pezzoli, 1980 plana, Hauffenia—Bole, 1961 planospira, “Fissuria”—Bodon, Cianfanelli & Talenti, 1997 polinskii, Horatia—Radoman, 1953 praeclara, Ногайа— Bourguignat, 1887 prlitchevi, Ohrigocea (Karevia)— HadZisce, 1959 pseudorientalica*, Islamia— Radoman, 1973a pusilla, Valvata—Piersanti, 1952 radapalladis, Pezzolia—Bodon & Giusti, 1986 raehlei, Horatia (Hauffenia)— Schutt, 1980 rotonda, Pseudamnicola (Rotondia)— Radoman, 1964 rudnicae, Hadziella— Bole, 1992 samuili, Ohrigocea (Ohrigocea)—Hadzisce, 1959 sandanskii, Ohrigocea (Karevia)— Hadzisce, 1959 schuelei, Hauffenia (Neohoratia) coronadoi— Boeters, 1981 servaini, Horatia— Bourguignat, 1887 sinjana, Hauffenia jadertina—Kuscer, 1933a sketi, Dalmatella— Velkovrh, 1970 sketi, Hadziella— Bole, 1961 sokolovi*, Horatia— Starobogatov, 1962 solitaria, Hauffenia— Bole & Velkovrh, 1986 spirata, Hauffenia minuta— Bernasconi, 1985 spiridoni, Bracenica— Radoman, 1973a st. naumi [sic], Pseudamnicola (Rotondia)— Radoman, 1964 Taxonomic status Hadziella krkae Kerkia kusceri Zaumia kusceri Pseudohoratia lacustris Islamia globulus lagari Islamia latina* junior synonym of Horatia klecakiana “Horatia” Iljovuschkini* junior synonym of Hauffenia kerschneri “Hauffenia” lucidula “Hauffenia” lucidula junior synonym of Ohrigocea stankovici Horatia macedonica Prespolitorea malaprespensis Hauffenia media junior synonym of Hauffenia tellinii possible junior synonym of /slamia globulina Islamia mienisi Ohrigocea miladinovorum nomen nudum Ohridohauffenia minuta Islamia minuta possible senior synonym of /s/amia globulina Daudebardiella naegelei* Horatia novoselensis junior synonym of Horatia klecakiana junior synonym of Horatia klecakiana Pseudohoratia ochridana Gocea ohridana Strugia ohridana Karevia ornata junior synonym of Horatia klecakiana “Daphniola” exigua pangaea “Horatia” parvula* Arganiella pescei “Hauffenia’ plana “Fissuria” planospira junior synonym of Pseudohoratia ochridana junior synonym of Horatia klecakiana junior synonym of Karevia omata Islamia pseudorientalica* Islamia pusilla Pezzolia radapalladis Fissuria raehlei Ohridohauffenia rotonda Hadziella rudnicae Ohrigocea samuili junior synonym of Karevia omata Islamia schuelei junior synonym of /slamia valvataeformis “Hauffenia” sinjana Dalmatella sketi Hadziella sketi “Horatia” sokolovi* nomen nudum Islamia spirata Bracenica spiridoni Ohridohauffenia sanctinaumi REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 77 TABLE 5. (Continued) Nominal taxon st. zaumi [sic], Horatia— Radoman, 1964 stankovici, Horatia (Hauffenia)—Hadzisce, 1959 stankovici, Lyhnidia—Hadzisce, 1959 sturmi, Paludina— Rosenhauer, 1856 subcarinata, Hauffenia (Hauffenia)— Bole & Velkovrh, 1987 sublitoralis, Lyhnidia— Radoman, 1967 sublitoralis, Pseudamnicola (Ohridohauffenia)— Radoman, 1963a subpiscinalis, Valvata (?) —KusScer, 1932 sulcata, Sardohoratia- Manganelli, Bodon, Cianfanelli, Talenti & Giusti, 1998 supracarinata, Horatia— Bole & Velkovrh, 1986 tellinii, Horatia (Hauffenia)—Pollonera, 1898 thermalis, Hadziella— Bole, 1992 tovunica, Hauffenia (Hauffenia)— Radoman, 1978 trichoniana*, Islamia— Radoman, 1979 troglobja*, “Valvata’— Bole 8 Velkovrh, 1986 turgidula, Valvata— Locard, 1889 umbilicata*, Haaziella—Bole & Velkovrh, 1986 valvataeformis, Horatia (Hauffenia)— Pollonera, 1898 valvataeformis, Hydrobia— Möllendorff, 1873 valvataeformis, Prespolitorea— Radoman, 1973a verdica, Erythropomatiana— Radoman, 1978 verlikana, Horatia— Bourguignat, 1887 wagneri, Valvata—Kuscer, 1928 wienerwaldensis, Hauffenia— Haase, 1992 zermanica, Islamia— Radoman, 1973a species appears to have a distal (or RS1) seminal receptacle (Boeters, 1988: 220, figs. 162, 174). Adistinct genus may have to be in- troduced for each of these species. However, we postpone this course of action until more anatomical detail is available. An even greater number of species was as- signed (Sometimes tentatively) to Neohoratia by Boeters (1988), Boeters & Rolan (1988), Rolan (1997a, b), and Hinz et al. (1994): N. ateni (Boeters, 1969), N. azarum Boeters & Rolan, 1988; N. (?) coronadoi (Bourguignat, 1870); N. (?) fezi (Altimira, 1960); N. (?) gasulli (Boeters, 1981); N. globulus (Bofill, 1909) (with two subspecies: N. g. globulus and N. g. lagari (Altimira, 1960)); and N. schuelei (Boeters, 1981). None of these ap- pear to belong to Hauffenia, as herein rede- fined. “Neohoratia” gasulli was more recently re- vised by Ramos et al. (1992) on topotypical material. It is peculiar in that it has a large pear-shaped bursa copulatrix and is com- pletely devoid of distinct seminal receptacles (according to Ramos et al., 1992, the function of seminal receptacles is supplied by the dis- Taxonomic status Zaumia sanctizaumi Ohrigocea stankovici Lyhnidia stankovici “Horatia” sturmi Hauffenia subcarinata Lyhnidia sublitoralis Ohridohauffenia sublitoralis Hauffenia subpiscinalis Sardohoratia sulcata nomen nudum (see Hauffenia subcarinata) Hauffenia tellinii Hadziella thermalis Hauffenia tovunica Islamia trichoniana* nomen nudum (= Islamia pusilla) possible junior synonym of /slamia globulina nomen nudum junior synonym of Hauffenia tellinii Islamia valvataeformis Prespolitorea valvataeformis junior synonym of Hauffenia subpiscinalis junior synonym of Horatia klecakiana Hauffenia wagneri Hauffenia wienerwaldensis Islamia zermanica tal portion of the renal oviduct which is en- larged and refringent, as if it contained ori- ented spermatozoa); the penis has a lobe. Also in this case, we think a distinct genus will probably have to be introduced. Anatomical study of some other species (“N.” ateni, “N.” g. globulus and “N.” g. lagari; Figs. 189-208), showed that: the female gen- italia are characterized by the absence of a bursa copulatrix and the presence of two rather relatively separated seminal recepta- cles, the proximal larger than the distal one (Figs. 194, 200, 206); the penis is character- ized by the presence of a large lobe on the left side containing a refringent mass of glandular cells; the penial duct runs inside the right side; an evident muscular pleat on ventral side, a glandular mass inside penis tip and a stylet- like structure at the opening of the penial duct are absent. Apart from the pecullar shell shape, amnicoliform or even bythinelliform (e.g., N. aten), the anatomical characters dis- tinguish these three taxa from those included in Hauffenia. They suggest that “N.” ateni, “N.” g. globulus and “N.” g. lagari belong to a group of /slamia, close to that including the 178 BODON, MANGANELLI & GIUSTI TABLE 6. Geographical distribution of valvatiform hydrobiid species from Europe and the Middle East (* taxa not discussed in the paper). The names of the countries are in parentheses when a species is present only in one or a few localities or areas of that country. RS Taxa Arganiella pescei Giusti & Pezzoli, 1980 Bracenica spiridoni Radoman, 1973a Dabriana bosniaca Radoman, 1974 Dalmatella sketi Velkovrh, 1970 Daphniola exigua exigua (Schmidt, 1856) “Daphniola” exigua pangaea (Reischútz, 1984) Daudebardiella asiana* Boettger, 1905 Daudebardiella naegelei* Boettger, 1905 Fissuria boui Boeters, 1981 Fissuria raehlei (Schútt, 1980) “Fissuria” planospira* Bodon et al., 1997 Gocea ohridana Hadzisce, 1956 Haaziella anti Schútt, 1960 Haaziella deminuta Bole, 1961 Haaziella ephippiostoma Kuscer, 1932 Haaziella krkae Bole, 1992 Haaziella rudnicae Bole, 1992 Haaziella sketi Bole, 1961 Haaziella thermalis Bole, 1992 Hauffenia danubialis (Haase, 1993) Hauffenia erythropomatia (Hauffen, 1856) Hauffenia kerschneri (Zimmermann, 1930) Hauffenia media Bole, 1961 Hauffenia subcarinata Bole & Velkovrh, 1987 Hauffenia subpiscinalis (Kuscer, 1932) Hauffenia tellinii (Pollonera, 1898) Hauffenia tovunica Radoman, 1978 Hauffenia wagneri (Kuscer, 1928) Hauffenia wienerwaldensis Haase, 1992 “Hauffenia” edlaueri (Schütt, 1961b) “Hauffenia” hadei (Gittenberger, 1982) “Hauffenia” jadertina KuScer, 1933a “Hauffenia” lucidula (Angelov, 1967) “Hauffenia” plana Bole, 1961 “Hauffenia” siniana KuScer, 1933a Heraultia exilis (Paladilhe, 1867) Horatia klecakiana Bourguignat, 1887 Horatia macedonica (Kuscer, 1936) Horatia novoselensis Radoman, 1966 “Horatia” birsteini* Starobogatov, 1962 “Horatia” borutzkii* Zhadin, 1932 “Horatia” gasulli (Boeters, 1981) “Horatia” gatoa Boeters, 1980 “Horatia” knorn* Schütt, 1961 “Horatia” ljovuschkini Starobogatov, 1962 “Horatia” parvula* (Naegele, 1894) “Horatia” sokolovi* Starobogatov, 1962 “Horatia” stumi (Rosenhauer, 1856) Islamia anatolica* Radoman, 1973a Islamia ateni (Boeters, 1969) Islamia azarum (Boeters & Rolan, 1988) Islamia bosniaca* Radoman, 1973a Islamia burnabasa* (Schütt, 1964) Islamia cianensis Bodon et al., 1995 Islamia consolationis (Bernasconi, 1985) Distribution Latium, Marche and Abruzzo (Italy) Montenegro Bosnia-Herzegovina Dalmatia (Croatia) Thessalia and Peloponnesus (Greece) East Makedonia (Greece) Turkey Turkey Vaucluse, Bouches du Rhóne, Var and Alpes Maritimes (France) Cephalonia and Zante (Greece) Tuscany (Italy) Lake Ohrid Rab Island (Croatia), Slovenia and Friuli (Italy) Friuli-Venetia Julia (Italy) and Slovenia Slovenia Slovenia Croatia Croatia Slovenia and Croatia Niederósterreich (Austria) Slovenia Oberósterreich and Niederósterreich (Austria) Slovenia and Croatia Slovenia Friuli-Venetia Julia (Italy) and Slovenia Eastern Venetia, Friuli-Venetia Julia (Italy) and Slovenia Croatia Slovenia Niederósterreich (Austria) Dalmatia Lakonia, Peloponnesus (Greece) Dalmatia (Croatia) Bulgaria Montenegro Dalmatia (Croatia) Hérault (France) Dalmatia (Croatia) and Bosnia-Herzegovina, Albania (?) Macedonia Macedonia Caucasus Caucasus East Spain South Spain Dalmatia Caucasus Turkey Caucasus South Spain Turkey Northeast Spain Northwest Spain Bosnia-Herzegovina Turkey Sicily (Italy) Doubs (France) REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 179 TABLE 6. (Continued) Taxa Distribution Islamia gaillardoti (Germain, 1911) Islamia gaiteri Bodon et al., 1995 Islamia globulina (Paladilhe, 1866) Islamia globulus globulus (Bofill, 1909) Islamia globulus lagari (Altimira, 1960) Islamia graeca* Radoman, 1973a Islamia latina* Radoman, 1973a Islamia mienisi (Schutt, 1991) Islamia minuta (Draparnaud, 1805) Islamia pseudorientalica* Radoman, 1973a Islamia pusilla* (Piersanti, 1952) Islamia schuelei (Boeters, 1981) Islamia spirata (Bernasconi, 1985) Islamia trichoniana* Radoman, 1979 Islamia valvataeformis (Móllendorff, 1873) Islamia zermanica Radoman, 1973a “Islamia” bendidis* Reischútz, 1988 “Islamia” coronadoi (Bourguignat, 1870) “Islamia” epirana (Schütt, 1962) “Islamia” fezi (Altimira, 1960) Karevia omata (Radoman, 1957) Kerkia brezicensis Bodon & Cianfanelli, 1996 Kerkia kusceri (Bole, 1961) Lyhnidia giorgievici Hadzisce, 1959 Lyhnidia hadzii Hadzisce, 1959 Lyhnidia karamani Hadzisce, 1959 Lyhnidia stankovici Hadzisce, 1959 Lyhnidia sublitoralis Radoman, 1967 Ohridohauffenia depressa (Radoman, 1957) Ohridohauffenia drimica (Radoman, 1964) Ohridohauffenia minuta (Radoman, 1955) Ohridohauffenia rotonda (Radoman, 1964) Ohridohauffenia sanctinaumi (Radoman, 1964) Ohridohauffenia sublitoralis (Radoman, 1963a) Ohrigocea karevi Hadzisce, 1959 Ohrigocea miladinovorum Hadzisce, 1959 Ohrigocea samuili Hadzisce, 1959 Ohrigocea stankovici (Hadzisce, 1959) Pezzolia radapalladis Bodon & Giusti, 1986 Prespolitorea malaprespensis Radoman, 1973a Prespolitorea valvataeformis Radoman, 1973a Pseudohoratia brusinae (Radoman, 1953) Pseudohoratia lacustris (Radoman, 1964) Pseudohoratia ochridana (Polinski, 1929) Pseudoislamia balcanica Radoman, 1979 Sardohoratia islamioides Manganelli et al., 1998 Sardohoratia sulcata Manganelli et al., 1998 Sheitanok amidicus* Schütt & Sesen, 1991 Strugia ohridana Radoman, 1973a Zaumia kusceri (HadZiSce, 1959) Zaumia sanctizaumi (Radoman, 1964) Israel Elba Island (Italy) South and East France Northeast Spain Northeast Spain Amvrakia Lake, Etolia (Greece) Dalmatia (Croatia) Israel and Lebanon Doubs, Jura and Ain (France), Neuchätel (Switzerland) Turkey Campania, Abruzzo and Apulia (Italy) South Spain Doubs (France) Trichonis Lake, Etolia (Greece) Bosnia-Herzegovina Croatia Samothraki Island (Greece) Central Spain Ipiros, Etolia and Lefkada (Greece) East Spain Lake Ohrid Slovenia Slovenia Sweti Naum Lake, Ohrid basin Lake Ohrid Lake Ohrid Lake Ohrid Lake Ohrid Lake Ohrid Ohrid basin Ohrid basin Lake Ohrid Ohrid basin Lake Ohrid Lake Ohrid Lake Ohrid Lake Ohrid Lake Ohrid and Ohrid basin Liguria (Italy) Mirka Prespa Lake (Albania) Prespa Lake Lake Ohrid Lake Ohrid Lake Ohrid Trichonis Lake Etolia (Greece) Sardinia Island (Italy) Sardinia Island (Italy) Turkey Ohrid basin Ohrid basin Lake Ohrid French “Hauffenia” species (see the re- description of the French Islamia in “Descrip- tions of some taxa misidentified as Hauffenia species’). North American Species. Two valvatiform species from North America, Valvata micra Pilsbry & Ferris, 1906, and Valvata micra nugax Pilsbry & Ferris, 1906, were assigned to Hauffenia by Bole & Velkovrh (1986) and to Hauffenia and Horatia respectively by Burch (1989). An anatomical revision by Hershler & Longley (1986) in the same years showed that these species belong to a distinct, North 180 BODON, MANGANELLI & GIUSTI TABLE 7. Analytical key for identification of the Hauffenia species (only species certainly belonging to Hauffenia are considered). For characters used, see Fig. 188. 1a—Operculum with peg, from small to very well developed 1b — Орегсшит without peg or, all the most, with very reduced peg 2a —Peg small, not dilated at the top 2b —Peg well developed, dilated at the top 3a —Shell with spire moderately to well raised; penis with one wide lateral lobe 3b —Shell with spire slightly raised 4a —Penis with one lateral lobe poorly developed; intestine with bend tightly coiled on pallial wall 4b —Penis with one wide lateral lobe; intestine with bend slightly coiled on pallial wall Ба — Peg of medium size; penis without or with slightly evident lobe/s 5b — Peg very well developed; penis with one rather evident lateral lobe 6a — Shell with a keel on the lower wall of the last whorl around umbilicus 6b — Shell not keeled 7a—Penis with 1-3 small lateral lobes; no peg 7b — Penis without lobes 8a — Shell with spire well raised; diameter 1.4-2.9 mm 8b — Shell with spire rather flat; diameter 1.17-1.55 mm 9a—No peg 9b — Peg very reduced . wagneri . media . kerschneri . tovunica . subcarinata . tellinii . subpiscinalis . erythropomatia . danubialis H. wienerwaldensis Dept ae <2 00) 270 American genus, Phreatodrobia Hershler & Longley, 1986. THE SPECIES OF HAUFFENIA Hauffenia danubialis (Haase, 1993) Lobaunia danubialis Haase, 1993: 99-105, figs. 8B, 9-15. Type Locality and Type Material: see Lobau- nia danubialis Haase, 1993, in the sec- tion on the taxa of the genus group. Diagnosis A species of Hauffenia having shell very small, valvatiform-planispiral, with spire al- most flat; operculum without peg; penis with- out lobes. Material Examined and Description See Lobaunia danubialis Haase, 1993, in the section on the taxa of the genus group. Distribution Niederósterreich, Austria. Taxonomy Hauffenia danubialis was assigned to a dis- tinct genus, Lobaunia, by Haase (1993). This genus is here regarded as a junior synonym of Hauffenia. It is distinct from H. wiener- waldensis, due to complete absence of an op- ercular peg. Hauffenia erythropomatia (Hauffen, 1856) Valvata erythropomatia Hauffen, 1856: 465. Type Locality and Type Material: see Eryth- ropomatiana erythropomatia (Hauffen, 1856) in the section devoted to the taxa of the genus group. Diagnosis A species of Hauffenia having shell very small, valvatiform, with spire rather flat; oper- culum without peg; penis with 2-3 small lat- eral lobes. Material Examined and Description See Erythropomatiana erythropomatia (Hauffen, 1856) in the section devoted to the taxa of the genus group. Distribution North of Ljubljana, Slovenia. Taxonomy Hauffenia erythropomatia was assigned to a distinct genus, Erythropomatiana, by Rado- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 181 4b 6b 8b 9a 9b FIG. 188. Characters used in the analytical key for the identification of the Hauffenia species (Table 7). man (1978). This genus is here regarded as a junior synonym of Hauffenia. Hauffenia kerschneri (Zimmermann, 1930) Ногайа erythropomatia kerschneri Zimmer- mann, 1930: 233-234, pl. 13, figs. 3-6. Type Locality: “. . . in den zufúhrenden Quell- kanalen der Wasserleitung in Weyer an der Enns (Oberósterreich)”, Austria. Type Material: lectotype (NHMW 85034) and 19 paralectotypes (NHMW K 48844) at the Naturhistorisches Museum Wien, Vi- enna, Austria (Haase, 1993). Hauffenia kerschneri loichiana Haase, 1993: 94-98. Type Locality: “Kleine Quelle unterhalb des Fuchslochs (Hóhlenkatastern. 1837/24) bei Loich im westlichen Niederósterreich (Seitental der Pielach)”, Austria. Type Material: holotype (NHMW 85029) and 68 paratypes (NHMW 85030, shells) at the Naturhistorisches Museum Wien, Vi- enna, Austria (Haase, 1993). Diagnosis A species of Hauffenia having shell very small, valvatiform, with spire not very raised; operculum with small peg; penis with one wide lateral lobe. Material Examined —Spring in Weyer an der Enns (type locality), Oberósterreich, Austria, M. Bodon leg. 10.6.1985 (3 males, 3 females, many shells). Description Shell very small, valvatiform, thin, waxen, transparent when fresh; surface of proto- conch malleated; spire not very raised, con- 182 BODON, MANGANELLI & GIUSTI 190 PGL 189 193 FIGS. 189-194. Shell, operculum and anatomical details of /slamia ateni (Boeters, 1969) from the thermal spring at Banys de Sant Vicieng, Cataluña, Spain, M. Bodon leg. 14.9.1985. Fig. 189: shell; Fig. 190: outer face of operculum; Fig. 191: body of a male with pallial cavity open to show head and penis; Fig. 192: dor- sal side and ventral side (second picture) of penis of three males; Fig. 193: prostate gland, intestine and pal- lial organs of a male; Fig. 194: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar = 1mm. sisting of 2.5-3.25 rather rapidly growing con- vex whorls; last whorl large, dilated, descend- ing slightly near aperture; umbilicus wide; aperture prosocline, oval; peristome com- plete, thin, slightly thickened, slightly reflected only at columellar margin (Figs. 214, 215; Zimmermann, 1930: 233-234, pl. 13, figs. 3-6; Haase, 1993: 92, fig. 1A-D, table 1, as Hauffenia k. kerschneri; Haase, 1993: 94, fig. 4A-D, table 1, as H. k. loichiana; Boeters, 1998: 28, fig. H6). Dimensions : height = 0.70-1.09 mm; diameter = 1.00-1.61 mm. Operculum thin, orange, paucispiral, with small spiralized peg at centre of inner face (Fig. 216; Haase, 1993: 92, fig. 2A, B, as Hauffenia k. kerschneri; Haase, 1993: 94, as H. k. loichiana). Body unpigmented (a few black spots on visceral sac); eye spots absent (Fig. 217; Haase, 1993: 94, as H. k. loichiana). Male genitalia with prostate gland bulging well into pallial cavity; penis short, flat, with apex blunt, and one lateral, not very raised but wide lobe on left side; penial duct zig-zag- ging through central portion of penis to open at penis tip; globular mass of refringent cells 204 FIGS. 195-206. Shell, operculum and anatomical details of Islamia globulus globulus (Bofill, 1909) from the spring near Guardiola, Cataluña, Spain, M. Bodon leg. 14.9.1985 (Figs. 195-200) and of /s/amia globulus lagari (Altimira, 1960) from the spring Fuente Les Dous, Torrelles de Foix, Cataluña, Spain, M. Bodon leg. 2.1.1992 (Figs. 201-206). Figs. 195, 201: shells; Figs. 196, 203: body of a male with pallial cavity open to show head and penis; Figs. 197, 202: outer face of operculum; Figs. 198, 205: prostate gland, stomach (ex- cluded in Fig. 198), intestine and pallial organs of a male; Figs. 199, 204: dorsal and ventral side (second picture) of penis; Figs. 200, 206: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar =1 mm. 184 BODON, MANGANELLI 8 GIUSTI FIGS. 207-213. Microsculpture of protoconchs. Fig. 207: Islamia ateni (Boeters, 1969) from the thermal spring at Banys de Sant Vicienc, Cataluña, Spain, M. Bodon leg. 14.9.1985; Fig. 208: Islamia globulus glob- ulus (Bofill, 1909) from the spring near Guardiola, Cataluña, Spain, M. Bodon leg. 14.9.1985; Fig. 209: Is- lamia minuta (Draparnaud, 1805) from the Source de l'Ain, Nozeroy, Jura, France, M. Bodon leg. 21.7.1985; Fig. 210: Islamia globulina (Paladilhe, 1866) from the Rivière souterraine de Labouiche, Foix, Ariège, France, M. Bodon leg. 13.9.1985; Fig. 211: Islamia spirata (Bernasconi, 1985) from the stream near Pont les Moulins, Cusancin valley, Doubs, France, M. Bodon leg. 22.7.1985; Fig. 212: Islamia consolationis (Bernasconi, 1985) from the springs at Consolation-Maisonnettes, Doubs, France, М. Bodon 8 G. Manganelli leg. 13.6.1996; Fig. 213: Heraultia exilis (Paladilhe, 1867) from the Source du Lez, Hérault, France, M. Bodon leg. 2.12.84. Scale bar = 100 um. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 185 214 FIGS. 214-221. Shell, operculum and anatomical details of Hauffenia kerschneri (Zimmermann, 1930) from the spring in Weyer an der Enns, Oberósterreich, Austria, M. Bodon leg. 10.6.1985. Figs. 214, 215: shells; Fig. 216: outer face (left), profile (centre) and inner face (right) of operculum; Fig. 217: body of a female with pallial cavity open to show head; Fig. 218: dorsal side of penis of two males; Fig. 219: prostate gland, stom- ach, intestine and pallial organs of a male; Fig. 220: renal and pallial oviduct, intestine and pallial organs of a female; Fig. 221: renal and pallial oviduct of a female. Scale bar = 1 mm. inside penis apex to right of penial duct (no data about this in Haase, 1993); terminal part of penial duct (immediately before opening) with very small stylet (Figs. 218, 219; Haase, 1993: 95-96, 98, figs. 5, 7, 8, as H. k. loichi- ana; Boeters, 1998: 28, fig. H7). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle very small, with very short duct arising from oviduct level with end of loop; bursa copulatrix re- duced, about same size as seminal recepta- cle, slightly dilated at apex, arising very close to where oviduct enters albumen gland por- tion of pallial oviduct; seminal groove running along ventral side of capsule gland (Figs. 220, 221; Haase, 1993: 95, 98, figs. 5, 6, as Hauf- fenia k. loichiana; Boeters, 1998: 28, fig. H8). Radula with central tooth trapezoidal with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 5 smaller denticles on both sides, in de- creasing order of size; 2 basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically en- larged, their anterior margin with 9 denticles, central one longer, larger; first marginal teeth rake-shaped, with long lateral wing and elon- 186 BODON, MANGANELLI & GIUSTI gated cutting edge with long row of 21-22 small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lat- eral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 13-15 very small denticles (Haase, 1993: 92, fig. 3A, B, as H. k. kerschneri; Haase, 1993: 94, fig. 3C, D, as H. k. loichiana). Stomach without posterior caecum; intes- tine with well developed, S-like bend on pallial wall (Figs. 219, 220; Haase, 1993: 95, fig. 5, as Н. к. loichiana). Osphradium variable in size, elliptical or kidney-shaped; ctenidium consisting of 6-9 lamellae (Figs. 219, 220; Haase, 1993: 94, as H. k. loichiana). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Haase, 1993: 95, as H. k. loichi- ana). Distribution Oberósterreich and Niederósterreich, Aus- tria. Taxonomy The small opercular peg and the penis with wide penial lobe suggest relationships with Hauffenia wagneri (KusScer, 1928) or H. media Bole, 1961. Nevertheless, due to other weak anatomical characters (intestine with less tightly coiled bend on pallial wall) and the dis- tance between the type localities and the dis- tributions of these species, we prefer to main- tain H. kerschneri as a distinct species. Haase (1993) divided the species into two subspecies on the basis of an unique differ- ence in the shell morphometry of a single pop- ulation; this is not significant considering the marked variability in shell dimensions of the single populations of Hauffenia species. Hauffenia media Bole, 1961 Hauffenia media Bole, 1961: 62-63, 67-68, fig. 3B. Type Locality: “jama Vrlovka pri Kamanju ob Kolpi”, Croatia. Type Material: Bole (1961) did not give any in- formation about the type material. Diagnosis A species of Hauffenia having shell very small, valvatiform, with spire slightly raised; operculum with very small peg; penis with one wide but not very raised lateral lobe. Material Examined —Spring near Kostanjevica, KrSko, Slovenia, 33T WL 37, A. Edlauer leg., (Naturhis- torisches Museum Wien no. 21418; 1 shell with operculum, 3 shells, deter- mined by H. Schütt). —“Kostanjeviska Jama’ cave, 5. 518, Kostan- jevica, Krsko, Slovenia, 33T WL 37, Е. Stoch leg. 16.6.1996 (1 female). Description Shell very small, valvatiform, thin, waxen, transparent when fresh; surface of proto- conch malleated; spire slightly raised, con- sisting of 3-3.25 rather rapidly growing con- vex whorls; last whorl large, dilated, descending slightly near aperture; umbilicus wide; aperture prosocline, roundish; peris- tome complete, thin, slightly reflected only at columellar margin (Figs. 222-224; Bole, 1961: 62-63, 67, fig. 3B). Dimensions: height = 1.0-1.2 mm; diameter = 1.4-1.9 mm (Bole, 1961). Operculum thin, paucispiral, yellowish, with very small spiralized peg at centre of inner face (Figs. 225, 226; Bole, 1961, 62, 67, fig. 3B; 1993: 6). Body unpigmented (a few traces of pigment in wall of visceral sac); eye spots absent (Fig. 227). Male genitalia with penis rather elongated, flat, with apex blunt, slightly pointed at centre and one wide, but not very raised lateral lobe on left side; penial duct zig-zagging through sub-central portion of penis to open at penis tip; no data available about existence of glob- ular mass of refringent cells inside penis apex to right of penial duct and of stylet (Bole, 1961: 62, 67, fig. 3B). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle small, with very short duct arising from oviduct level with end of loop; bursa copulatrix reduced, a little longer than seminal receptacle, slightly dilated at apex, arising very close to where oviduct enters albumen gland portion of pallial oviduct; seminal groove running along ventral side of capsule gland (Fig. 228). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 187 FIGS. 222-228. Shell, operculum and anatomical details of Hauffenia media Bole, 1961, from the spring near Kostanjevica, KrSko, Slovenia, A. Edlauer leg., (Naturhistorisches Museum Wien no. 21418) (Figs. 222, 223, 225) and from the “Kostanjeviska Jama” cave, S. 518, Kostanjevica, KrSko, Slovenia, F. Stoch leg. 16.6.1996 (Figs. 224, 226-228). Figs. 222-224: shells; Figs. 225, 226: outer face (left), profile (centre) and inner face (right) of operculum; Fig. 227: body of a female with pallial cavity open to show head; Fig. 228: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar = 1 mm. and 4-5 smaller denticles on both sides in de- creasing order of size; one basal cusp where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically en- larged, their anterior margin with 10 denticles, central one longer, larger; first marginal teeth with apical row of 23-28 denticles; second marginal teeth with apical row of 18-24 denti- cles (Bole, 1961: 62, 67, fig. 3B). Stomach without posterior caecum; intes- tine with well-developed, tightly coiled, S-like bend on pallial wall (Fig. 228). 188 BODON, MANGANELLI & GIUSTI Osphradium elongated; ctenidium consist- ing of 8-12 lamellae (Fig. 228; Bole, 1961: 62, 67). Nervous system unknown. Distribution Eastern Slovenia and northern Croatia. Taxonomy The anatomical data is not complete enough to draw conclusions on the status of this species. Penis shape, female genitalia and operculum structure are sufficient to infer it belongs to Hauffenia. The small opercular peg suggests relationships with H. wagneri (Kuscer, 1928). Hauffenia subcarinata Bole & Velkovrh, 1987 Horatia supracarinata Bole & Velkovrh, 1986: 193, nomen nudum. Hauffenia (Hauffenia) subcarinata Bole & Velkovrh, 1987: 72-73, 78-79, fig. 1A-C, pl. 1, figs. 1, 2. Type Locality: “Izvir v vasi Lozice pri Desklah v dolini Soce. 12 km severno severoza- hodno od Nove Gorice/Quelle im Dorf Lozice bei Deskle in Soca-Tal, 12 km NNW von Gorica”, Slovenia. Type Material: holotype (35215a) in the Velkovrh collection, BiotehniSka fakulteta Univerza Edvarda Kardelja, Ljubljana, Slovenia, together with paratypes (35215/80) (Bole & Velkovrh, 1987). Diagnosis A species of Hauffenia having shell very small, valvatiform, with spire moderately raised; lower wall of last whorl with keel; op- erculum with well-developed peg; penis with- out lobes. Material Examined —Spring near Lozice, along road to Kanal, Soca valley, Slovenia, 33T UM 9201, M. Bodon leg. 10.7.1996 (1 shell with oper- culum, many shells) (type locality). —Spring in camping area at Kanal, Soca val- ley, Slovenia, 33T UM 9404, S. Cian- fanelli & M. Calcagno leg. 31.7.1994 (1 shell with operculum, 5 shells). Description Shell very small, valvatiform, pale whitish, waxen, transparent when fresh; surface of protoconch malleated; spire moderately raised, conical, consisting of 2.75-3.5 rather rapidly growing convex whorls; last whorl large, dilated, descending slightly near aper- ture, its lower wall (around umbilicus) having keel ending at aperture; umbilicus wide; aper- ture prosocline, oval, slightly angled at lower margin (near keel); peristome complete, thin, slightly thickened, slightly reflected only at lower and columellar margin (Figs. 94, 229; Bole & Velkovrh, 1987: 72, 78, pl. 1, figs. 1, 2; Bole, 1993: 6). Dimensions: height = 0.83- 1.30 mm; diameter = 1.14-1.90 mm (Bole & Velkovrh, 1987). Operculum thick, yellowish brown, pau- cispiral, with well-developed, spiralized peg at centre of inner face (Fig. 230; Bole, & Velkovrh, 1987: 72, 79, fig. 1B, C; Bole, 1993: 6). Body unpigmented; eye spots absent. Male genitalia with penis rather elongated, cylindrical, flat, slightly tapering near apex, ending in a slightly blunt tip, without penial lobes; penial duct zig-zagging through central portion of penis to open at penis tip; no data available about existence of globular mass of refringent cells inside penis apex to right of penial duct and of stylet (Bole & Velkovrh, 1987: 72, 79, fig. 1A; Bole, 1993: 6). Female genitalia unknown. Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 4 smaller denticles on both sides in de- creasing order of size; two basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically en- larged, their anterior margin with 10 denticles, central one longer, larger; first marginal teeth rake-shaped, with long lateral wing and elon- gated cutting edge with long row of 18-20 small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lat- eral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 14-16 very small denticles (Bole & Velkovrh, 1987: 72, 79, fig. 1C; Bole, 1993: 6). Stomach, intestine, osphradium, ctenidium and nervous system unknown. Distribution Soéa [Isonzo] valley, western Slovenia. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 189 Taxonomy Penis characters and operculum with peg are probably sufficient to justify inclusion of this species in the genus Hauffenia. НаиНета subpiscinalis (Ки5бег, 1932) Valvata (?) subpiscinalis Kuscer, 1932: 5-53, 21: 5, 19.1. Type Locality and Type Material: see Neoho- ratia subpiscinalis (Kuscer, 1932) in the section devoted to the taxa of the genus group. Erythropomatiana verdica Radoman, 1978: 36, pl. 5, figs. 20, 21. Type Locality and Type Material: see the syn- onymy of Neohoratia subpiscinalis (Kuscer, 1932) in the section devoted to the taxa of the genus group. Diagnosis A species of Hauffenia having shell very small but larger than the other species of the genus, valvatiform, with spire well raised; op- erculum without peg; penis with 1-3 small lat- eral lobes. Material Examined and Description See Neohoratia subpiscinalis (KuScer, 1932) in the section devoted to the taxa of the genus group. Distribution Eastern Friuli-Venetia Julia, Italy, and West- ern Slovenia, from Soca [Isonzo] valley to Ljubljana area. Taxonomy Hauffenia subpiscinalis was recently as- signed to a distinct genus, Neohoratia, by Bole & Velkovrh (1986), Boeters (1988), and Bole (1993). This genus is here regarded as a junior synonym of Hauffenia. Erythropomatiana verdica Radoman, 1978, is here recognized as a junior synonym of H. subpiscinalis. Radoman (1978) distinguished H. verdica from H. subpiscinalis on the basis of few differences (penis shape; slightly larger bursa copulatrix), deduced from the drawings of H. subpiscinalis published by Bole (1967: fig. 3A). It is true that Bole showed a squatter penis with larger lobes than those figured by Radoman (1978: fig. 6) in H. erythropomatia, but as we verified with topotypical specimens, E. verdica also has a squat penis (Fig. 130). Hauffenia subpiscinalis from the typical area showed penis lobes variable in number (2-3) and shape, often very similar to those of E. verdica and H. erythropomatia (Figs. 138, 140). Regarding the bursa copulatrix, our studies on many populations not only re- vealed bursa size to be rather variable, but also that specimens of H. subpiscinalis from the typical area have a bursa proportionally larger than those figured by Bole (1967) and similar to those found in E. verdica from Vrhnika (Figs. 131, 135-137). Shell shape is also similar in the type species of the two gen- era (cf., Bole, 1970, 1979; Radoman, 1978). Hauffenia tellinii (Pollonera, 1898) Horatia (Hauffenia) Tellinii Pollonera, 1898: 3, 4, fig. 2. Type Locality and Type Material: see Hauffe- nia tellinii (Pollonera, 1898) in section de- voted to taxa of the genus group. Horatia (Hauffenia) valvataeformis Pollonera, 1898: 3-4, fig. 3. Type Locality and Type Material: see syn- onymy of Hauffenia tellinii (Pollonera, 1898) in section devoted to the taxa of the genus group. Hauffenia michleri KuScer, 1932: 56-57, pl. 5, fig. 3. Type Locality and Type Material: see syn- onymy of Hauffenia tellinii (Pollonera, 1898) in section devoted to the taxa of the genus group. Diagnosis A species of Hauffenia having shell very small, valvatiform to planispiral, with spire from rather raised to almost flat; operculum with well developed peg; penis with 1-2 slightly evident lateral lobes, sometimes ab- sent. Material Examined and Description See Hauffenia tellinii (Pollonera, 1898) in section devoted to taxa of the genus group. 190 BODON, MANGANELLI & GIUSTI Distribution From eastern Venetia and Friuli-Venetia Julia, Italy, to western Slovenia in the Ljubl- jana area. Taxonomy We agree with Bole (1970) about the syn- onymy of the two species described by Pollonera from the upper Natisone valley: Ho- ratia (Hauffenia) tellinii and H. (H.) valvatae- formis. The latter is distinguished from the for- mer by a shell with raised spire, which is clearly only one end of a spectrum of shell shape. The two extreme morphs (spire de- pressed: tellinii; spire raised: valvataeformis) and their intermediates were found in many of the populations examined. Our study does not confirm the existence of constant and valid shell, anatomical and op- ercular characters that justify regarding H. michleri as a species distinct from H. tellinii. Traditional distinction of the two species was evidently based on anatomical study of very few specimens. Hence, we propose H. mich- leri as a junior synonym of H. tellinii. Hauffenia populations from the southeast- ern Karst in the province of Trieste have an operculum with a reduced peg; they were ten- tatively assigned to H. tellinii (Pezzoli, 1988a; Bodon & Giovannelli, 1993). Hauffenia tovunica Radoman, 1978 Hauffenia (Hauffenia) tovunica Radoman, 1978: 34, fig 5A-E, pl. 4, figs. 11, 12. Type Locality: “Die Grotte Tounjcica, neben dem Ort Tounj, in der Náhe der Strasse Duga resa — Josipdol”, Croatia. Type Material: holotype and four paratypes (SMF 249614) at the Senckenberg-Mu- seum, Frankfurt am Main, Germany (Jo- vanovic, 1991). Diagnosis A species of Hauffenia having shell very small, valvatiform, with spire not very raised; operculum with very well-developed dilated peg; penis with one rather evident lateral lobe. Material Examined —“Tounjcica Spilja” cave, Tounj, east of Ogulin, Croatia, E. Kletecki, F. Gasparo & F. Stoch leg. 13.7.1997 (1 male, 1 young specimen, 12 shells). Description Shell very small, valvatiform, not very thin, transparent when fresh; surface of proto- conch malleated; spire not very raised, con- sisting of 3.25-3.75 rather rapidly growing convex whorls; last whorl large, dilated, slightly descending near aperture; umbilicus wide; aperture prosocline, roundish; peris- tome complete, somewhat thickened and re- flected (Fig. 231; Radoman, 1978: 34, pl. 4, figs. 11, 12; Radoman, 1983: 122, pl. 9, fig. 142; Jovanovic, 1991: pl. 8, fig. 66). Dimen- sions: height = 1.26-1.68 mm; diameter = 1.81-2.02 mm (Radoman, 1983: table 7). Operculum rather thick but with thin edge, paucispiral, convex, but concave at centre, with very well-developed, dilated and spiral- ized peg at centre of inner face (Fig. 235; Radoman, 1978: 33-34, fig. 5D, E; Radoman, 1983: 120, fig. 67D, E). Body unpigmented (a few black spots on visceral sac); eye spots absent (Fig. 233). Male genitalia with prostate gland bulging well into pallial cavity; penis rather short, flat, with apex blunt, and one rather evident, knob- like, lateral lobe on left side near apex; penial duct zig-zagging through central portion of penis to open at penis tip; globular mass of re- fringent cells inside penis apex to right of pe- nial duct and terminal part of penial duct with very small stylet (Fig. 232; Radoman, 1978: 33, fig. 5C; 1983: 120, fig. 67C). Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle very small, with very short duct arising from oviduct level with end of loop; bursa copulatrix re- duced, small but markedly longer than semi- nal receptacle, slightly dilated at apex, arising very close to where oviduct enters albumen gland portion of pallial oviduct; seminal groove running along ventral side of capsule gland (Radoman, 1978: 33, fig. 5A, B; 1983: 40, 120, fig. 67A, B). Radula with central tooth with one pair of basal cusps; other details unknown (Rado- man, 1978: 33; 1983: 114). Stomach without posterior caecum; intes- tine with well-developed, tightly coiled, S-like bend on pallial wall (Fig. 234; Radoman, 1983: 40). Osphradium kidney-shaped; ctenidium consisting of about 11 lamellae (Fig. 234). FEN ASS oP airs) = 236 FIGS. 229-236. Shell, operculum and anatomical details of Hauffenia subcarinata Bole & Velkovrh, 1987, from the spring near LoZice, along the road to Kanal, Soëa valley, Slovenia, 33T UM 9201, M. Bodon leg. 10.7.1996 (Figs. 229, 230), Hauffenia tovunica Radoman, 1978, from the “Tounjéica Spilja” cave, Tounj, east of Ogulin, Croatia, E. Kletecki, F. Gasparo 8 F. Stoch leg. 13.7.1997 (Figs. 231-235) and of Hauffenia wienerwaldensis Haase, 1992, from the upper well in Klamm 106, Wienerwald, Niederósterreich, Austria, M. Haase leg. 17.7.1989 (Fig. 236). Figs. 229, 231, 236: shells; Figs. 230, 235: outer face (left), profile (centre) and inner face (right) of operculum; Fig. 232: dorsal side of penis; Fig. 233: body of a male with pallial cav- ity open to show head and penis; Fig. 234: prostate gland, stomach, intestine and pallial organs of a male. Scale bar = 1 mm. 192 BODON, MANGANELLI & GIUSTI Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Radoman, 1978: 33; 1983: 120). Distribution Known only from the type locality, near Ogulin, northern Croatia. Taxonomy The study of the male genitalia confirms that the this species is distinct and that it be- longs to Hauffenia. Compared to H. tellinii, H. tovunica has more developed opercular peg and penial lobe. Hauffenia wagneri (KuScer, 1928) Valvata wagneri KuScer, 1928: 50, fig. 1. Type Locality and Type Material: see Vrania wagneri (Kuscer, 1928) in section de- voted to taxa of the genus group. Diagnosis A species of Hauffenia having shell very small, conical-valvatiform or valvatiform, with spire from moderately to well raised; opercu- lum with small peg; penis with one wide lateral lobe. Material Examined and Description See Vrania wagneri (Kuscer, 1928) in sec- tion devoted to taxa of the genus group. Distribution Mirna valley, eastern Slovenia. Taxonomy The small opercular peg suggests relation- ships with Hauffenia media Bole, 1961. The variability of shell shape suggests that the two taxa may be synonyms. We prefer to maintain them as distinct species, pending further anatomical study of H. media. Hauffenia wienerwaldensis Haase, 1992 Hauffenia wienerwaldensis Haase, 1992: 208-213, figs. 1-11, table 1. Type Locality: “Upper well in Klamm 106”, Wienerwald, Niederósterreich, Austria. Type Material: holotype (NHMW 85940) and paratypes (NHMW 85941-85946) at the Naturhistorisches Museum Wien, Vi- enna, Austria; other paratypes are in the Reischútz collection (Baden, Austria) (Haase, 1992). Diagnosis A species of Hauffenia having shell very small, valvatiform, with spire from almost flat to slightly raised; operculum with very re- duced peg; penis without lobes. Material Examined —Upper well in Klamm 106, Wienerwald, Niederósterreich, Austria, M. Haase leg. 17.7.1989 (13 shells). Description Shell very small, valvatiform, thin, waxen, transparent when fresh; surface of proto- conch malleated; spire from almost flat to slightly raised, consisting of 3-3.25 rather rapidly growing convex whorls; last whorl large, dilated, slightly descending near aper- ture; umbilicus wide; aperture prosocline, oval; peristome complete, thin, slightly re- flected only at columellar margin, sometimes detached from last whorl (Fig. 236; Haase, 1992: 208, figs. 1, 2; Boeters, 1998: 29, figs. H9, 10). Dimensions: height = 0.67-1.00 mm; diameter = 1.15-1.69 mm (Haase, 1992: table 1). Operculum thin, orange, paucispiral, slightly thickened at centre of inner face to give rise to very reduced peg (Haase, 1992: 208, fig. 3A-D). Body unpigmented (a few black spots on visceral sac); eye spots absent (Haase, 1992: 208, fig. 4). Male genitalia with penis rather short, large, flat, slightly tapering near apex, ending in a slightly blunt tip and without penial lobes; pe- nial duct zig-zagging through central portion of penis to open near penis tip; no data avail- able about existence of globular mass of re- fringent cells inside penis apex to right of pe- nial duct; terminal part of penial duct (immediately before opening) with well-devel- oped stylet, which according to original de- scription “stands somewhat behind the tip of penis perpendicular to its axis and slightly in- clined to the right” (Haase, 1992: 210, figs. 9-11; Boeters, 1998: 29, fig. H11). REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 193 Female genitalia with proximal seminal re- ceptacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle very small, sessile or with very short duct arising from oviduct level with end of loop; bursa cop- ulatrix reduced, same size as seminal recep- tacle, slightly dilated at apex, arising very close to where oviduct enters albumen gland portion of pallial oviduct; seminal groove run- ning along ventral side of capsule gland (Haase, 1992: 209-210, fig. 8; Boeters, 1998: 29, fig. H12). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with long robust central denticle and 5 smaller denticles on both sides in de- creasing order of size; 1-2 basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically en- larged, their anterior margin with 10-12 denti- cles, central one longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 19-21 small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lat- eral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 15-17 very small denticles (Haase, 1992: 208, fig. 6A, B). Stomach without posterior caecum; intes- tine with well-developed, S-like bend on pallial wall (Haase, 1992: 208-209, fig. 7). Haase (1992: 208) reports an “anterior digestive gland lobe” with a separate opening into the stomach. Osphradium kidney-shaped; ctenidium consisting of 9-10 lamellae; Haase (1992: 208) reports the presence of a hypobranchial gland (Haase, 1992: 208, fig. 5). Nervous system with long pleuro-supraoe- sophageal and short pleuro-suboesophageal connectives (Haase, 1992: 209). Distribution Niederósterreich, Austria. Taxonomy The published data clearly confirms that the present species belongs to Hauffenia. As for the hypobranchial gland and the anterior di- gestive gland-lobe (the latter found by Haase, 1992, in only 25% of dissected specimens), two structures never described before in the group (and which must be more carefully in- vestigated), their meaning for taxonomy ap- pears presently irrelevant. DESCRIPTIONS OF SOME TAXA MISIDENTIFIED AS HAUFFENIA SPECIES Fissuria raehlei (Schutt, 1980) Horatia (Hauffenia) raehlei Schutt, 1980: 140. Type Locality: “Insel Kephallinia, Schlucht bei Poros, Genist des Baches”, Greece. Type Material: holotype (SMF 263529) at the Senckenberg-Museum, Frankfurt am Main, Germany (Schütt, 1980). Material Examined — Well no. G/54, Cephalonia, Greece, С. |. Pesce, D. Maggi & M. Miranda leg. 7.5.1977 (1 male, 1 female) (Pesce et al., 1979). —Well no. G/57, S. Efimia, Cephalonia, Greece, G. L. Pesce, D. Maggi & M. Mi- randa leg. 7.5.1977 (1 male, 1 female) (Pesce et al., 1979). — Well no. G/58, $. Efimia, Cephalonia, Greece, G. L. Pesce, D. Maggi & M. Mi- randa leg. 7.5.1977 (2 males, 1 shell) (Pesce et al., 1979). — Well no. G/143, Poros, Cephalonia, Greece, G. L. Pesce, D. Maggi & G. Silverii leg. 2.4.1978 (1 male) (Pesce et al., 1979). —Well no. G. 174, shore Zante-Lithakial, at the crossroads for Mouzaki, Zante, Greece, G. L. Pesce & G. Silverii leg. 8.4.1979 (1 male, 5 females, 2 shells) (Pesce & Maggi, 1983). —Well no. G. 194, near Katastarion, Zante, Greece, G. L. Pesce & G. Silverii leg. 9.4.1979 (1 male, 1 female, 2 shells) (Pesce & Maggi, 1983). Description Shell very small, valvatiform-planispiral, thin, whitish, transparent when fresh; surface of protoconch malleated; spire slightly raised, consisting of 2.75-3.25 rather rapidly grow- ing, convex whorls; last whorl large, dilated, descending near aperture; umbilicus very wide; aperture prosocline, roundish-ovoid; peristome complete, slightly reflected only at columellar margin (Figs. 237-238; Schutt, 1980: 140, pl. 10a, fig. 42). Dimensions: height = 0.54-1.07 mm; diameter = 1.11-1.52 mm. Operculum thin, yellowish, paucispiral, 194 BODON, MANGANELLI & GIUSTI FIGS. 237-249. Shell, operculum and anatomical details of Fissuria raehlei (Schutt, 1980) from well no. G/58, S. Efimia, Cephalonia, Greece, G. L. Pesce, D. Maggi & M. Miranda leg. 7.5.1977 (Figs. 237, 239, 241-242, well no. G/174, shore Zante-Lithakial, at the crossroads for Mouzaki, Zante, Greece, G. L. Pesce & G. Silverii leg. 8.4.1979 (Figs. 238, 247, 248), well no. G/54, Cephalonia, Greece, G. L. Pesce, D. Maggi & M. Miranda leg. 7.5.1977 (Figs. 240, 243, 244), well no. G/57, S. Efimia, Cephalonia, Greece, G. L. Pesce, D. Maggi & M. Miranda leg. 7.5.1977 (Figs. 245, 249) and well no. G/194, near Katastarion, Zante, Greece, G. L. Pesce & G. Silverii leg. 9.4.1979 (Fig. 246). Figs. 237-238: shells; Fig. 239: outer face (left) and pro- file (centre) of operculum; Fig. 240: body of a male with pallial cavity open to show head and penis; Figs. 241, 244-247: dorsal side of penis of six males; Fig. 242: prostate gland, intestine and pallial organs of a male; Fig. 243: renal and pallial oviduct, intestine and pallial organs of a female; Figs. 248, 249: renal and pallial oviduct of two females. Scale bar = 1 mm. slightly thickened at centre of inner face but without peg (Fig. 239). Body unpigmented; eye spots absent (Fig. 240). Male genitalia with prostate gland bulging well into pallial cavity; penis rather short or moderately long, flat, with apex pointed, and two, well-raised lobes; lobes of variable size and containing mass of glandular tissue: one on left side about 2/3 of penis length; another on dorsal-right side about 1/3 of penis length; penial duct zig-zagging through right portion of penis to open at penis tip (Figs. 241, 242, 244-247). Female genitalia with two seminal recepta- cles and a bursa copulatrix arising from distal renal oviduct; proximal and distal seminal re- ceptacles elongated, more or less equal in size; bursa copulatrix variable in size from medium to very large, roundish to oval, with slender elongated duct entering bursa on an- terior side; seminal groove running along ven- tral side of capsule gland (Figs. 243, 248, 249). Radula with central tooth trapezoidal with long lateral wings and basal tongue; two basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 10-11 denticles, central one longer, larger; REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 195 first marginal teeth rake-shaped, with long lat- eral wing and elongated cutting edge with long row of 28-32 small denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of about 22 very small denti- cles (Eigs: 172, 173). Stomach without posterior caecum; intes- tine with rather developed, S-like bend on pal- lial wall (Figs. 242, 243). Osphradium variable in size, elongated, oval or kidney-shaped; ctenidium consisting of 9-23 lamellae (Figs. 242, 243). Nervous system unknown. Taxonomy The study of many specimens from Cephalonia Island (some collected close to the type locality) and Zante Island revealed that the female genitalia has medium to large bursa copulatrix and two rather long seminal receptacles (proximal and distal) and that the male genitalia have a penis with two glan- dular lobes. These characters agree with those described for Fissuria boui and conse- quently support inclusion of Horatia (Hauffe- nia) raehlei in the same genus. Islamia minuta (Draparnaud, 1805) Valvata minuta Draparnaud, 1805: 42, pl. 1, figs. 36-38. Type Locality: no locality is indicated by Dra- parnaud (1805) or is given in the labels accompanying the type material in the Draparnaud collection at Naturhisto- risches Museum Wien (K. Edlinger, pers. com.). It must therefore be assumed that the type locality is “France” from the title of Draparnaud's volume (“Histoire na- turelle des mollusques terrestres et fluvi- atiles de la France”). If the designation of a neotype is ruled by the ICZN, the type locality will become Source de l'Ain, Nozeroy, Jura. Type Material: original type material consists of the unidentifiable lectotype (a shell) in the Draparnaud collection, Naturhisto- risches Museum Wien (Austria) (Binder, 1966). Bernasconi (1975) selected a neotype [“nouveau typoide”] for Valvata minuta, but this designation is invalid (see below). In order to preserve this name in the current sense, we are apply- ing to the ICNZ to set aside the type sta- tus of the lectotype and to designate the shell shown in Fig. 250 as neotype. The proposed neotype is in the Naturhis- torisches Museum Wien (catalogue no. 100485). Material Examined —Source de l'Ain, Nozeroy, Jura, M. Bodon leg. 21.7.1985 (3 males, 1 female, many shells). —Source de ГЕртайег, near Poncin, Ain, M. Bodon leg. 26.8.1989 (1 male, 1 female, 39 shells). —Source du Dessoubre, Consolation-Mai- sionnettes, Doubs, M. Bodon leg. 21.7.1985 (1 male, 4 females, 48 shells). — Springs at Consolation-Maisonnettes, Doubs, M. Bodon & G. Manganelli leg. 13.6.1996 (6 males, 14 females, many shells). — Springs along the stream in the Parc du Seminaire, N.D. de Consolation, Conso- lation-Maisonnettes, Doubs, M. Bodon leg. 21.7.1985 (6 males, 7 females, 27 shells). — Source de l’Aiguille, Vallon Pont d'Arc, Ardèche, M. Bodon leg. 25.6.1989 (5 males, 3 females, many shells). This population, assigned by Bernasconi (1988) to /. globulina, is here reported as Islamia cf. minuta because anatomically is similar to /. minuta, but the shell spire is more conical, like /. globulina. Description Shell very small, almost planispiral, thin, whitish, waxen, transparent when fresh; sur- face of protoconch malleated; spire almost flat, consisting of 2.75-3.25 rather rapidly growing, convex whorls: last whorl large, di- lated, descending slightly near aperture; um- bilicus wide; aperture prosocline, roundish; peristome complete, thin, slightly thickened, slightly reflected only at columellar margin (Figs. 209, 250-251; Draparnaud, 1805: 42, pl. 1 figs. 36-38; Locard, 1889: 331-333; 1893: 128; Germain, 1931: 674-675; Binder, 1966: 371-374, figs. 1, 2; Bernasconi, 1975: 308-309, figs. 7a, b, 8a; 1977: 30, figs. 2a, 3a; Boeters, 1998: 28, figs. H1, 2). Dimen- sions: height = 0.67-1.56 mm; diameter = 1.09-2.11 mm (height = 0.7-1.5 mm; diame- ter = 1.0-2.2 mm according to Bernasconi, 1975 1977); Operculum thin, yellow orange, paucispiral, 196 BODON, MANGANELLI & GIUSTI FIG. 250. Proposed neotype of Islamia minuta (Draparnaud, 1805) from the Source de l'Ain, Nozeroy, Jura, M. Bodon leg. 21.7.1985. Naturhistorisches Museum Wien (catalogue no. 100485). Scale bar = 1 mm. a little thicker at centre but without outgrowth on inner face (Fig. 257; Bernasconi, 1975: 307, fig. 6b, c). Body unpigmented (sometimes traces of pigment in wall of visceral sac); eye spots present or absent (Figs. 253, 255; Boeters, 1979: 60; Bernasconi, 1975: 304, fig. 1b). Male genitalia with prostate gland bulging Slightly into pallial cavity; penis rather elon- gated, flat, with sides corrugated near base, not or slightly tapering near apex, which branches in two; right branch corresponding to tip of penis proper, variably elongated, cylin- drical or conical, pointed; left branch being a lobe, variable in shape and size, but always shorter than penis tip and with inside refringent mass of glandular cells; rather straight muscu- lar pleat on ventral side about 2/3 of penis length, running obliquely from right side to base of penial lobe and not protruding on left side; penial duct zig-zagging through right or central portion of penis to open at penis tip (Figs. 252, 254, 256; Bernasconi, 1975: 305-306, fig. 4b, c; Boeters, 1998: 28, fig. H4). Female genitalia with only two seminal re- ceptacles arising from distal renal oviduct, usually not very close to one another; that arising at end of loop (in position correspond- ing to that of proximal) well developed, always larger and longer than other, with short but ev- ident stalk, wider at apex; that arising close to where oviduct enters albumen gland (in posi- tion more or less corresponding to that of dis- tal) very small, usually without evident stalk, in some specimens with refringent mass of ori- ented spermatozoa; no trace of bursa copula- trix; seminal groove running along ventral side of capsule gland (Figs. 259-261; Bernasconi, 1975: 306, fig. 5b; Boeters, 1998: 28, fig. H5). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with larger central denticle and five smaller denticles on both sides in de- creasing order of size; 2-3 basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically en- larged, their anterior margin with 11-13 denti- cles, central one longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 20-26 FIGS. 251-261. Shell, operculum and anatomical details of Islamia minuta (Draparnaud, 1805) from the Source de ГАт, Nozeroy, Jura, M. Bodon leg. 21.7.1985 (Figs. 251 -253, 255, 257, 259), from the springs along the stream in the Parc du Seminaire, N.D. de Consolation, Consolation-Maisonnettes, Doubs, M. Bodon leg. 21.7.1985 (Figs. 254, 261), and from the Source de l'Epinglier, near Poncin, Ain, M. Bodon leg. 26.8.1989 (Figs. 256, 260). Fig. 251: shell; Figs. 252, 254, 256: dorsal and ventral side (the second picture of each series) of penis of seven males; Fig. 253: body of a male with pallial cavity open to show head and penis; Fig. 255: head of another specimen with eye spots; Fig. 257: outer face (left) and profile (right) of op- erculum; Fig. 258: prostate gland, stomach, intestine and pallial organs of a male; Figs. 259, 260: renal and pallial oviduct, intestine and pallial organs of two females; Fig. 261: renal and pallial oviduct of a female. Scale bar = 1 mm. 198 BODON, MANGANELLI & GIUSTI H/D 08 % H/D 0.6 204 104 262 06 08 10 H/D 263 10 12 14 16 D H H/D 10 H/D 0.8 % 1.2 р H/D 06 204 10 ES Y 10) 0.8 Ve er 264 06 08 10 H/D 265 10 12 14 16 D FIGS. 262-265. Biometric analysis of the shells of some populations of the French /slamia. Figs. 262, 263: /. minuta (Draparnaud, 1805) and /. globulina (Paladilhe, 1866) from the Source de l’Epinglier, near Poncin, Ain, M. Bodon leg. 26.8.1989; Figs. 264, 265: /. minuta and /. spirata (Bernasconi, 1985) from the springs along the stream in the Parc du Seminaire, N.D. de Consolation, Consolation-Maisonnettes, Doubs, M. Bodon leg. 21.7.1985. H: shell height; D: shell diameter; dimensions in mm. small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lat- eral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 14-18 very small denticles (Bernasconi, 1975: 304-305, fig. 2b, c). Stomach without posterior caecum; intes- tine with well-developed, S-like bend on pallial wall (Figs. 258-260; Bernasconi, 1975: 306, fig. 3b; Boeters, 1998: 28, fig. H3). Osphradium oval; ctenidium consisting of 8-14 lamellae (Figs. 258-260; Bernasconi, 1975: 305, fig. 3b). Nervous system unknown. Distribution The distribution of /slamia in France in- cludes a large part of the Rhone basin to the east, the Languedoc and the Garonne, Dor- dogne basins and probably the Gascogne (Fig. 185) to the southwest. The distribution indicated by Bouchet (1990: Fig. 7) for “Hauf- fenia” minuta and by Ripert (1998: fig. 10) for “Neohoratia” globulina includes eastern Provence and the Cóte d'Azur, southeastern areas in which /slamia has never been docu- mented (anatomical study of valvatiform hy- drobiids from the Départements of Vaucluse, Var and Alpes Maritimes, identified them as Fissuria boui Boeters, 1982). Populations surely attributed to /. minuta have only been found in the Jura mountains, the upper Rhone basin, the French départ- ments of Doubs, Jura and Ain and the Swiss cantons of Neuchatel and Bern (Bernasconi, 1975, 1977). Taxonomy Our study shows that the relative position of the two seminal receptacles and the point where they arise from the renal oviduct are quite constant in all the populations examined REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 199 of Islamia minuta, 1. globulina (Paladilhe, 1866), /. spirata (Bernasconi, 1975) and /. consolationis (Bernasconi, 1985) (for the last three, see Descriptions below). It is important that the proximal seminal receptacle is always larger and longer than the distal one (see Description of /slamia). This contrasts with what Bernasconi (1975, 1977, 1984, 1985) claimed, that is, distal sac-like structure (aris- ing from renal oviduct just before it enters pal- lial oviduct) larger than proximal (arising close to end of loop). The fact that at least part of the material studied by Bernasconi comes from the same localities as the material stud- ied by us (Source de l’Ain, Nozeroy for /. min- uta; Vidourle, resurgence, Sauve for /. globu- lina) indicates that our material cannot belong to a taxon different from Bernasconi’s and suggests that he misinterpreted the female genital structure. This may have been be- cause in wrongly interpreting the larger sac-like structure as a bursa copulatrix, Bernasconi illustrated it arising where the bursa copulatrix normally arises in the hydro- biids, that is, from the renal oviduct immedi- ately before the oviduct enters the pallial oviduct. This misinterpretation is indirectly confirmed by the fact that the position of the smaller sac-like structure, interpreted by Bernasconi as a seminal receptacle, is also imprecisely figured by him. In Bernasconi’s figures, it does not have a constant position and sometimes arises proximally from the renal oviduct at the end of the loop and some- times about halfway along the loop (see Bernasconi, 1975, fig. 5; 1977, fig. 1e; 1984, fig. 1e; 1985, fig. 1d). Our data on the structure of the female gen- italia and the penis (presence of glandular lobe on left side of penis; absence of mass of refringent cells inside penis apex to right of penial duct; absence of stylet at tip of penis) is enough to conclude that these French species do not belong to Hauffenia, as Haase (1993: 106) suggested. The misinterpreted French “Hauffenia” must be assigned to /s- lamia. In fact, they only differ from the type species of /slamia from the Balkans (to which species from Italy, Asia Minor and Middle East are more or less related; cf., Radoman, 1973a, 1983; Giusti & Pezzoli, 1980; Giusti et al., 1981; Schutt, 1991; Bodon et al. 1995a) in some minor anatomical features: for example, seminal receptacles sometimes more distant from one another and relatively undeveloped muscular pleat on ventral side of penis which does not protude on left side. It is particularly difficult to clarify the identity of Valvata minuta and the status of the forms/subspecies into which it has been sub- divided. Valvata minuta is cited by many clas- sic authors from various localities (Férussac, 1807: 128; Bouchard-Chantereaux, 1838: 87; Gassies, 1849: 183; 1859: 60; Dupuy, 1851: 585-586; Moquin-Tandon, 1856: 434). Pal- adilhe (1866: 25, 27) claimed that Gassies and the others misinterpreted it as “une char- mante Valvée microscopique de forme glob- uleuse”, which he described as a distinct species: V. globulina. Locard (1895: 20, 46-47) traced the type material of V. minuta. This consists of two shells in the Draparnaud collection in the Naturhistorisches Museum Wien, Austria, and one in the M. Bischof von Hohenwarth collection. Many years later, Binder (1966: 371-372) studied the two syntypes in the Naturhistorisches Museum Wien. One is a hydrobiid species (and is se- lected as the Чуре”), the other is a fragment of the apex of a shell of Valvata piscinalis (Muller, 1774). Bernasconi (1975) published a first re- description of V. minuta based on the study of some Swiss and French populations, but his paper contains many controversial aspects which have made a puzzle of the case. First of all, although he was aware of the existence of a lectotype, Bernasconi selected a neotype [“nouveau typoide”] for У minuta, in contra- vention of ICZN (1999: Art. 75) and conse- quently invalid. Moreover, his neotype does not belong to the nominotypical form/sub- species minuta but to the different form/sub- species globulina. The type locality indicated for the form/subspecies globulina (Vidourle, résurgence (Sauve)), and not that of the form/subspecies minuta (Areuse, résurgence (St. Sulpice)), is reported as type locality of the species. Both these restricted type locali- ties are outside the range reported by the au- thors of the taxa. In fact, that of the form/sub- species minuta is in the Swiss canton of Neuchátel (under the king of Prussia until 1815) and not in France; that of form/sub- species globulina in the Rhone basin, and not in the Garonne basin. As a consequence, his neotype designation and his type locality re- striction are invalid. Study of the lectotype (Draparnaud collec- tion no. 1820 xxvi/21; Naturhistorisches Mu- seum Wien, Austria; Binder, 1966: fig. 1) does not enable certain identification of the spe- cies. It has a shell shape similar to the speci- mens of /. minuta from the Jura, but the shell 200 BODON, MANGANELLI & GIUSTI TABLE 8. Shell parameters of the /slamia species coexisting in the Source de ГЕртойег at Poncin, Ain, and the springs in the Parc du Seminaire at Consolation-Maisonnettes, Doubs. Acronyms: H shell height, D shell diameter. Locality Source de l'Epinglier, Poncin (Ain) mean ratio H/D + o (range) number of shells Springs in the Parc du Seminaire, Consolation-Maisonnettes (Doubs) mean ratio H/D + o (range) number of shells height is less. On the basis of shell size (height: 0.60 mm; diameter: 1.34 mm), it is more similar to Valvata exilis Paladilhe, 1867, from the Départment de ГНегаий (see “De- scription of a new valvatiform genus from France” for redescription of V. exilis), differing in the fact that the last whorl is not dilated near the aperture. No locality is indicated by Dra- parnaud (1805), nor is it given in the labels ac- companying the type material in the Dra- parnaud collection. If Draparnaud, who lived in Montpellier, collected this material near his town, then his V. minuta cannot be the species that is currently considered to be V. minuta, which lives much further north, but it may be V. exilis. In this situation, we think that it is better to apply to ICZN to set aside the Binder’s (1966) type designation and to des- ignate a neotype for this species so that the current understanding of this nominal taxon is preserved. The proposed neotype (Fig. 250) was col- lected in the Source de ГАт, Nozeroy, Jura. This proposed neotype (a shell) was chosen from a population for which the anatomy is al- ready known (Bernasconi, 1975). This popu- lation lives in a major spring of the French Jura. No other similar /slamia species lives in the same spring. The neotype is deposited in the Naturhistorisches Museum Wien (cata- logue no. 100485). Bernasconi (1984, 1985, 1988) regarded /. minuta as having four subspecies. All of them, apart from /. consolationis, which is character- ized by a larger shell, are identified by different values of the ratio of shell height and diameter (H/D) (/. minuta: 0.56 + 0.07-0.66 + 0.09; /. globulina: 0.76 + 0.09-0.91 + 0.06; /. spirata: 1.00 + 0.06-1.07 + 0.08; Bernasconi, 1975, 1985). In cases of coexistence of more forms/ Taxa Islamia minuta Islamia globulina 0.65 + 0.05 0.93 + 0.06 (0.51-0.73) (0.82-1.06) Sil 28 Islamia minuta Islamia spirata 0.62 + 0.06 0.98 + 0.04 (0.53-0.81) (0.91-1.06) 30 19 subspecies, shell material clearly revealed that the populations are not homogeneous and that the frequencies of H/D values are dis- tributed on a bimodal curve. For example, the population from Source de l'Epinglier, Ain, consists of two quite distinct shell morphs (Table 8; Figs. 262, 263). Binder (1966) stud- ied the shells and claimed that the depressed shells, traditionally assigned to V. minuta, and the shells with raised spire, traditionally as- signed to V. globulina, were linked by a series of intermediate shells, and were the extremes of a continuum. He concluded that V. minuta and V. globulina were not distinct species (“Cette distinction ne se justifie probablement pas. . .”). The evidence of two distinct coex- isting morphs is undeniable in this population. Unfortunately, there is too little anatomical data, only one female and one male (Fig. 256) examined, to confirm their status as distinct taxa on an anatomical basis. lf we are to ac- cept the H/D criteria of Bernasconi (1975, 1985), these two morphs can be assigned to /. minuta and to /. globulina. The same happens near Consolation- Maisonnettes, Doubs, where three distinct shell morphs also coexist (Table 8; Figs. 264, 265). Again, according to the dimensions and the H/D values of Bernasconi (1975, 1985), the three morphs can be assigned to /. mi- nuta, to |. spirata and to I. consolationis. Anatomical study revealed that the speci- mens attributed to /. minuta and /. consolatio- nis have a penial lobe shorter than the penial apex, and those assigned to /. spirata a penial lobe longer than the penial apex, while only /. minuta has eye spots (Figs. 254, 276-277, 282, 284). Other specimens such as those collected in the Source de l’Aiguille, Vallon Pont d’Arc, REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 201 Ardeche, and assigned by Bernasconi (1988) to /. globulina are characterized by H/D values (mean H/D: = 0.76 + 0.05), which lie almost between those of /. globulina and I. minuta whereas the male genitalia, are identical to those of /. minuta (penial lobe shorter than penis apex). Islamia spirata is readily distinguishable by shell parameters only in the case of the type population. When other populations are con- sidered the distinction becomes problemati- cal. As well, the absence of eyes in the type population of /. spirata is of no help for distin- guishing it from /. globulina. In fact, other pop- ulations assignable to /. minuta or /. globulina contained specimens with eyes coexisting with specimens without eyes, suggesting that this character is occasionally variable. The above facts are evidence of a very complex situation and possibly of equivocal shell and anatomical characters. However, since almost all these taxa (except /. globulina and /. spirata) live together in some places and are distinguishable by morphometric and anatomical characters, there must be at least three distinct species. More populations must obviously be studied (especially from the type localities of all the taxa described in the past) in order to gather more biometric and anatom- ical data for a better understanding of the tax- onomic setting of the group. We nevertheless feel justified in rejecting the current interpreta- tion of all French /slamia as subspecies of /. minuta. We suggest provisional recognition of the four taxa, /. minuta (Draparnaud, 1805), /. globulina (Paladilhe, 1866), /. spirata (Bern- asconi, 1975), and /. consolationis (Bern- asconi, 1985) as distinct species (see key in Table 9, Fig. 266). The shells of the different species of /slamia described to date (Radoman, 1973a, 1983; Giusti 4 Pezzoli, 1980; Giusti et al., 1981; Schütt, 1991; Bodon et al., 1995a) vary little in shape (mostly valvatiform with the spire raised to different degrees) and size. These two char- acters are not sufficiently diagnostic to deter- mine the status of allopatric taxa. This obvi- ously compels us to rely on anatomy, which also offers little in the way of diagnostic char- acters. Apart from the fact that French /slamia colonize an area geographically separate from the other areas inhabited by this genus (Spain, Italy, Corsica, Elba, Sicily, Balkans, Turkey, and Israel), two characters seem to confirm their distinctiveness. These characters, if con- stant, would be sufficient to support the exis- tence of a distinct branch in the radiation of Islamia. In fact, the French /slamia are distin- guished from all the eastern species, from the Balkans to Turkey and Israel (Radoman, 1973a, 1983; Giusti et al., 1981; Schutt, 1991), by seminal receptacles, on the average, more distant from one another and a relatively un- developed muscular pleat on ventral side of penis, not protruding on left side as in typical Islamia. The same differences distinguish the French /slamia from the Italian /. pusilla (Pier- santi, 1952) (Giusti et al., 1981), while a markedly reduced penial lobe distinguishes two other recently described /slamia species TABLE 9. Provisional analytical key for the identification of French Islamia species. For characters used, see Fig. 266. 1a — Penial lobe shorter than penis apex 2 1b — Penial lobe longer than penis apex 3 2a —Mean D < 2 mm; mean H/D = 0.56-0.66 I. minuta 2b —Mean D > 2 mm; mean H/D = 0.80-0.90 |. consolationis За — Mean H/D = 0.76-0.93 I. globulina 3b —Mean H/D = 0.98-1.07 |. spirata 2a 2b 3a 3b FIG. 266. Characters used in the analytical key for the identification of the French /s/amia species (Table 9). 202 BODON, MANGANELLI 4 GIUSTI from different Italian islands: /. cianensis Bodon, Manganelli, Sparacio & Giusti, 1995 (penis with muscular pleat), and /. gaiteri Bodon, Manganelli, Sparacio & Giusti, 1995 (penis without muscular pleat; Bodon et al., 1995a). Finally, other undescribed or little known Italian (see Giusti et al., 1981), Corsi- can and Spanish taxa differ in other anatomi- cal characters and/or the frequent absence of a muscular pleat on the ventral side of penis (for the Spanish species, see above). Islamia globulina (Paladilhe, 1866) ? Valvata moquiniana Dupuy, 1851: 586-587, pl. 28, fig. 15. Type Locality: “. . . alluvions du Lot, pres de Mende. . .”, Dep. Lozère, France. Type Material: Dupuy (1851) did not give any information about the type material. Valvata globulina Paladilhe, 1866: 170 [p. 27 in reprint]. Type Locality: “. . . paraît spéciale au bassin de la Garonne, notamment dans les dé- partements du Gers et de Lot-et- Garonne.”, France. Type Material: Paladilhe (1866) did not give any information about the type material; Binder (1966) stated that the type does not exist. ?Valvata bourguignati Letourneux, 1869: 197-199. Type Locality: “. . . fontaine, pres du Moulin- Gachet (commune de Pissotte)”, Dep. Vendée, France. Type Material: Letourneux (1869) did not give any information about the type material. ? Valvata turgidula Locard, 1889: 333-334. Type Locality: “. . . lac de la Négresse, pres de Bayonne, dans les Basses-Pyrénées с ЕГАЙСЕ. Type Material: according to Locard (1889) type material is in the Bourguignat collec- tion, Geneve, Switzerland. ?Valvata micrometrica Locard, 1889: 336- 337. Type Locality: “. . . fontaine du Camarde, pres de Valence dans le Gers”, France. Type Material: Locard (1889) did not give any information about the type material. Material Examined —Source de l’Epinglier, near Poncin, Ain, M. Bodon leg. 26.8.1989 (34 shells). —Vidourle, resurgence, Sauve, Gard, 1.1976, ex R. Bernasconi collection (2 males, 2 females); M. Bodon leg. 2.12.1984 (3 shells). —Source de la Nizon, Lirac, Gard, M. Bodon, H. Girardi & B. Bomba leg. 29.12.1998 (3 males, 1 female, many specimens, many shells). —Wells 13/PT/2, propriété Michon, Chateau- renard, Bouches du Rhone, C. Bou leg. 4.1977, collection H. Girardi (1 male). —Resurgence du Moulis at Moulis, St. Girons, Ariege, M. Bodon leg. 13.9.1985 (1 male, 2 females, 1 shell). —Riviere souterraine de Labouiche, Foix, Ar- iege, M. Bodon leg. 13.9.1985 (1 male, 2 females, many shells). —Ruisseau souterrain d’Amiel, Penne, Tarn, C. Bou leg. 12.1991, ex H. Girardi collec- tion (3 males, 1 female). —Ruisseau souterrain de Cabeau, Penne, Tarn, C. Bou leg. 12.1976, H. Girardi col- lection (1 male, 1 female). —Source de la Mandre, Soreze, Tarn, C. Bou leg. 10.1991, H. Girardi collection (1 male, 6 shells). —Water-bearing stratum of the intake A.E.P. of Lescure, Tarn, C. Bou leg. 12.1991, ex H. Girardi collection (5 males, 2 females). —Fontaine des Canelles, Lalinde, Dordogne, M. Bodon leg. 27.6.1989 (1 female). This population is reported as /slamia cf. glo- bulina, because there were no male specimens for anatomical study. Description Shell very small, valvatiform, thin, whitish, waxen, transparent when fresh; surface of protoconch malleated; spire rather raised, consisting of 2.5-3.25 rather rapidly growing, convex whorls; last whorl large, dilated, de- scending slightly near aperture; umbilicus small; aperture prosocline, roundish; peris- tome complete, thin, slightly thickened, slightly reflected only at columellar margin (Figs. 210, 267; Gassies, 1849: 183, pl. 2, fig. 7, as V. min- uta; Moquin-Tandon, 1855: 543, pl. 41, figs. 26-28, as V. minuta; Locard, 1889: 334-336, 1893: 127-128, fig. 130; Dupuy, 1851: 585, pl. 28, fig. 14, as V. minuta; Germain, 1931: 675-676, figs. 742, 743; Binder, 1966: 371-374, figs. 2, 3; Bernasconi, 1975: 308, 310, figs. 7c, 8b; 1977: 30, figs. 2a, 3a; 1984: 205, fig. 7f). Dimensions: height = 0.7-1.8 mm; diameter = 0.9-1.9 mm (height and di- ameter according to Bernasconi, 1975, 1977). Operculum thin, pale yellowish or yellow, REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 203 paucispiral, a little thicker at centre but without outgrowth on inner face (Fig. 268; Boeters, 1973: 65; Bernasconi, 1975: 307, fig. 6a; 1984: fig. 1b). Body unpigmented (sometimes traces of pigment in wall of visceral sac); eye spots present or absent (Fig. 269; Boeters, 1973: 65; Bernasconi, 1975: 304, fig. 1a; 1984: fig. 1a). Male genitalia with prostate gland bulging slightly into pallial cavity; penis variably elon- gated, flat, with sides corrugated near base, not or slightly tapering near apex, which branches in two: right branch, forming tip of penis proper, short, conical, more or less pointed; left branch constituting lobe, variable in shape and size but always slightly longer than penis tip, with inside refringent mass of glandular cells; rather straight muscular pleat on ventral side about 2/3 of penis length, run- ning obliquely from right side to base of penial lobe, not protruding on left side; penial duct zig-zagging through right or central portion of penis to open at penis tip (Figs. 270, 271; Boeters, 1973: 65; Bernasconi, 1975: 305- 306, fig. 4a; 1984: fig. 1a, d). Female genitalia with only two seminal re- ceptacles arising from distal renal oviduct, usually not very close to one another; that aris- ing at end of loop (in position corresponding to that of proximal) well developed, always larger and longer than other, with short but evident stalk and apical enlargement; that arising close to where oviduct enters albumen gland (in position more or less corresponding to that of distal) very small, usually without evident stalk; no trace of bursa copulatrix; seminal groove running along ventral side of capsule gland (Figs. 272, 273; Boeters, 1973: 65; Bernasconi, 1975: 306, fig. 5a; 1984: fig. 1e). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like, with larger central denticle and 5 smaller denticles on both sides, in decreas- ing order of size; 2-3 basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 11-13 denticles, central one longer, larger; first marginal teeth rake- shaped, with long lateral wing and elongated cutting edge with long row of 20-26 small denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lateral wing and roundish, spoon-like apex, its cut- ting edge carrying rather long row of 14-18 very small denticles (Bernasconi, 1975: 304-305, fig. 2a). Stomach without posterior caecum; intes- tine with well developed, U- or S-like bend on pallial wall (Figs. 272, 274; Boeters, 1973: 65; Bernasconi, 1975: 306-307, fig. 3a). Osphradium variable in size, oval or elon- gated; ctenidium consisting of 6-11 lamellae (Figs. 272, 274; Boeters, 1973: 65; Bern- asconi, 1975: 305, fig. 3a; 1984: fig. 1c). Nervous system unknown. Distribution The distribution of /. globulina includes much of France: the Rhone basin to the east, and the Languedoc, the Garonne and Dor- dogne basins and probably the Gascogne to the southwest (Bernasconi, 1975, 1977; Bertrand, 1998; Ripert, 1998). Taxonomy See also discussion of /. minuta. All the /s- lamia populations from southern France hav- ing shell characters of /. globulina can be con- sidered to belong to the same species on the basis of anatomical characters. Although, it has not been studied topotypi- cal material of all nominal species, having globulina-like shell described from southern France (e.g.,Valvata moquiniana Dupuy, 1851; V. globulina Paladilhe, 1866; V. bour- guignati Letourneux, 1869; V. turgidula Lo- card, 1889; and V. micrometrica Locard, 1889), it is very probable that they are syn- onymous. If this is true, the oldest established name available for this taxon is V. moquini- ana. Here we have maintained its current name V. globulina, awaiting a definitive revi- sion, including the study of material from the Départments of Gers and Lot-et-Garonne, the type locality of the species described by Pal- adilhe (1866). Islamia spirata (Bernasconi, 1985) Hauffenia minuta spirata Bernasconi, 1985: 63-64, fig. 1a-e, table 1. Type Locality: “Bléfond, résurgence (Silley; V. 3)”, Dép. Doubs, France. Type Material: Bernasconi (1975) did not give any information about the type material. Material Examined —Springs along the stream in the Parc du Seminaire, N.D. de Consolation, Conso- 204 BODON, MANGANELLI & GIUSTI 268 FIGS. 267-280. Shell, operculum and anatomical details of /slamia globulina (Paladilhe, 1866) from Vi- dourle, resurgence, Sauve, Gard, France 1.1976, ex R. Bernasconi collection (Figs. 267, 268, 271-272, 274) and from the Résurgence du Moulis at Moulis, St. Girons, Ariege, France, M. Bodon leg. 13.9.1985 (Figs. 269, 270, 273) and of Islamia spirata (Bernasconi, 1985) from the springs along the stream in the Parc du Seminaire, N.D. de Consolation, Consolation-Maisonnettes, Doubs, France, M. Bodon leg. 21.7.1985 (Figs. 275-280). Figs. 267, 275: shells; Figs. 268, 278: outer face (left) and profile (right) of operculum; Figs. 269: body of a male with pallial cavity open to show head and penis; Figs. 270, 271, 277: dorsal and ventral side (second picture of each series) of penis of four males; Figs. 272, 279: renal and pallial oviduct, intestine and pallial organs of a female; Fig. 273: renal and pallial oviduct of a female; Figs. 274, 280: prostate gland, stom- ach, intestine and pallial organs of a male; Fig. 276: body of a female with pallial cavity open to show head. Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 205 lation-Maisonnettes, Doubs, France, M. Bodon leg. 21.7.1985 (4 males, 7 fe- males, 9 shells). —Stream near Pont les Moulins, Cusancin valley, Doubs, France, M. Bodon leg. 22.7.1985 (many shells). Description Shell very small, valvatiform, thin, whitish, waxen, transparent when fresh; surface of protoconch malleated; spire raised, consist- ing of 3-3.5 rather rapidly growing, convex whorls; last whorl large, dilated, descending slightly near aperture; umbilicus small; aper- ture prosocline, roundish; peristome com- plete, thin, slightly thickened, slightly reflected only at columellar margin (Figs. 211, 275; Bernasconi, 1975: 308, 310, figs. 7d, 8c; 1985: table 1, fig. 2). Dimensions: height = 0.90-1.51 mm; diameter = 0.90-1.44 mm (height = 1.0-1.9 mm; diameter = 1.0-1.8 mm, according to Bernasconi, 1975). Operculum thin, yellowish, paucispiral, a lit- tle thicker at centre but without outgrowth on inner face (Fig. 278; Bernasconi, 1985: 63, fig. 1a). Body unpigmented (sometimes traces of pigment in wall of visceral sac); eye spots ab- sent (Fig. 276; Bernasconi, 1985: 63). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather elon- gated, flat, with sides corrugated near base, tapering slightly near apex, which branches into two; right branch, forming tip of penis proper, short, conical, more or less pointed; left branch, constituting lobe, cylindrical, slightly longer than penis tip, with inside re- fringent mass of glandular cells; rather straight muscular pleat on ventral side about 2/3 of penis length, running obliquely from right side to base of penial lobe, not protrud- ing on left side; penial duct zig-zagging through right or central portion of penis to open at penis tip (Fig. 277; Bernasconi, 1985: 63-64, fig. 1e). Female genitalia with only two seminal re- ceptacles arising from distal renal oviduct, rather close to one another: that arising at end of loop (in position corresponding to that of proximal) well developed, always larger, longer than other, with short but evident stalk and apical enlargement; that arising close to where oviduct enters albumen gland (in posi- tion more or less corresponding to that of dis- tal) very small, usually without evident stalk; no trace of bursa copulatrix; seminal groove running along ventral side of capsule gland (Fig. 279; Bernasconi, 1985: 64, fig. 1d). Radula details unknown (Bernasconi, 1985: 63). Stomach without posterior caecum; intes- tine with well-developed, S-like bend on pallial wall (Figs. 279, 280; Bernasconi, 1985: 64, Па. 15,6): Osphradium variable in size, oval or elon- gated; ctenidium consisting of 3-10 lamellae (Figs. 279, 280; Bernasconi, 1985: 63, fig. 1b). Nervous system unknown. Distribution The distribution of /. spirata is limited to a small area in the Jura mountains, upper Rhone basin, in the French Départment of Doubs (Bernasconi, 1975, 1985). Taxonomy See discussion of /. minuta. Islamia consolationis (Bernasconi, 1985) Hauffenia minuta consolationis Bernasconi, 1985: 64, figs. 2, 3, table 2. Type Locality: “Maurepos, . . . bei Consola- tion-Maisonettes,” Doubs, France. Type Material: holotype at Museum d'Histoire Naturelle de Geneve, Geneva, Switzer- land; paratypes are in the Bernasconi collection, Múnchenbuchsee, Switzer- land (Bernasconi, 1985). Material Examined —Grotte du Biez-Airoux, Consolation-Maison- nettes, Doubs, M. Bodon & G. Manganelli leg. 13.6.1996 (3 males, 2 females, 10 shells). —Grotte de Maurepos, Consolation-Maison- nettes, Doubs, M. Bodon & G. Manganelli leg. 13.6.1996 (1 shell). — Source du Dessoubre, Consolation-Maison- nettes, Doubs, M. Bodon leg. 21.7.1985 (many shells), M. Bodon & G. Manganelli leg. 13.6.1996 (2 females, 17 shells). —Springs at Consolation-Maisonnettes, Doubs, M. Bodon & G. Manganelli leg. 13.6.1996 (many shells). —Springs along the stream in the Parc du Seminaire, N.D. de Consolation, Conso- 206 BODON, MANGANELLI & GIUSTI 281 PSR FIGS. 281-286. Shell, operculum and anatomical details of /slamia consolationis (Bernasconi, 1985) from the Grotte du Biez-Airoux, Consolation-Maisonnettes, Doubs, France, M. Bodon & G. Manganelli leg. 13.6.1996. Fig. 281: shell; Fig. 282: body of a male with pallial cavity open to show head and penis; Fig. 283: outer face (left) and profile (right) of operculum; Fig. 284: dorsal and ventral side (second picture) of penis of two males; Fig. 285: testis, vas efferens, prostate gland, stomach, intestine and pallial organs of a male; Fig. 286: renal and pallial oviduct, intestine and pallial organs of a female. Scale bar = 1 mm. REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 207 lation-Maisonnettes, Doubs, M. Bodon leg. 21.7.1985 (3 shells). Description Shell very small, valvatiform, thin, whitish, waxen, transparent when fresh; surface of protoconch very finely malleated; spire rather raised, consisting of 2.75-3.5 rather rapidly growing convex whorls; last whorl large, di- lated, descending slightly near aperture; um- bilicus small; aperture prosocline, roundish; peristome complete, thin, slightly thickened, slightly reflected only at columellar margin (Figs. 212, 281; Bernasconi, 1985: table 2, fig. 3). Dimensions: height = 1.80-2.80 mm; di- ameter = 1.94-2.79 mm (height =1.63 + 0.53-2.09 + 0.15 mm; diameter = 2.01 + 0.36-2.32 + 0.21 mm according to Bernasconi, 1985). Operculum thin, yellow orange, paucispiral, a little thicker at centre but without outgrowth on inner face (Fig. 283). Body unpigmented (traces of pigment in wall of visceral sac); eye spots usually absent (Fig. 282). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather elon- gated, flat, with sides corrugated near base, not or slightly tapering near apex, which branches in two; right branch, forming tip of penis proper, variably elongated, conical, more or less pointed; left branch constituting lobe, cylindrical, shorter than penis tip, with in- side refringent mass of glandular cells; rather straight muscular pleat present on ventral side at about 3/4 the penis length, running obliquely from right side to base of penial lobe, not protruding on left side; penial duct zig-zagging through right or central portion of penis to open at penis tip (Fig. 284). Female genitalia with only two seminal re- ceptacles arising from distal renal oviduct, rather close to one another: that arising at end of loop (in position corresponding to that of proximal) well developed, always larger, longer than other, with short but evident stalk and apical enlargement; that arising close to where oviduct enters albumen gland (in posi- tion more or less corresponding to that of dis- tal) small, without evident stalk; no trace of bursa copulatrix; seminal groove running along ventral side of capsule gland (Fig. 286). Radula with central tooth trapezoidal, with long lateral wings and basal tongue, its apical margin V-like with long, robust central denticle and 5 smaller denticles on both sides in de- creasing order of size; 1-2 basal cusps where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically en- larged, their anterior margin with 9 denticles, central one longer, larger; first marginal teeth rake-shaped, with long lateral wing and elon- gated cutting edge with long row of 15-17 long denticles anteriorly; second marginal teeth scraper-shaped, with long, slender lat- eral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of about 15 small denticles (Figs. 174-176). Stomach without posterior caecum; intes- tine with well-developed, S-like bend on pallial wall (Figs. 285, 286). Osphradium variable in size, oval or elon- gated; ctenidium consisting of 10-15 lamellae (Figs. 285, 286). Nervous system unknown. Distribution The distribution of /. consolationis is limited to a small area in the Jura mountains, upper Rhone basin, in the French Départment of Doubs (Bernasconi, 1985). Taxonomy See discussion of /. minuta. DESCRIPTION OF A NEW VALVATIFORM GENUS FROM FRANCE Some of the species mentioned in the pres- ent paper and studied anatomically in the course of its preparation were found to belong to genera which have not yet been described. Here we only describe one, the type species of which was already known in the literature under an improper generic name. Heraultia n. gen. Diagnosis Hydrobiid snail with shell very small, plani- spiral, whitish, anatomically characterized by: male genitalia with penis conical, pointed, without lobes; female genitalia with distal seminal receptacle and a bursa copulatrix; bursa copulatrix very large, with very long 208 BODON, MANGANELLI & GIUSTI 292 FIGS. 287-294. Shell, operculum and anatomical details of Heraultia exilis (Paladilhe, 1867) from the spring of the Lez River, Hérault, France, M. Bodon leg. 2.12.84. Fig. 287: shell; Fig. 288: outer face (left) and pro- file (right) of operculum; Fig. 289: body of a male with pallial cavity open to show head and penis; Fig. 290: dorsal side of penis of three males; Fig. 291: prostate gland, stomach, intestine and pallial organs of a male; Fig. 292: renal and pallial oviduct, intestine and pallial organs of a female; Figs. 293-294: renal and pallial oviduct of two females. Scale bar = 1 mm. duct entering bursa on lateroposterior side; central tooth of radula with one basal cusp on each side. Etymology From Hérault, the Départment of France, in which the species occurs. Type Species Valvata exilis Paladilhe, 1867. Taxonomy See discussion of H. exilis. Heraultia exilis (Paladilhe, 1867) Valvata exilis Paladilhe, 1867: 51-52, pl. 21, figs. 27-30. Type Locality: “.. . dans les fossés d'irrigation des prairies de la rive droit du Lez a la hauter du hameau de Lattes .. . Elle se rencontre aussi en petit quantité dans les alluvions du Lez recueillies sous le vil- lage de Castelnau, a pres de 8 kilometres en dessus de Lattes. Mais elle parait bien plus abondante dans les alluvions de la Boyne (sous le village de Fontes, ar- rondissement de Béziers) . . .”, Hérault, France. Type Material: syntypes (shells) in the Pal- REVISION OF EUROPEAN VALVATIFORM HYDROBIIDS 209 adilhe collection, Faculté des Sciences, Montpellier, France (Boeters, 1974). Material Examined —Source du Lez, Hérault, France, M. Bodon leg. 2.12.1984 (3 males, 5 females, many shells), 4.9.1985 (many shells). —Debris of the Lez River, Prades-le-Lez, Hérault, France, M. Bodon leg. 5.1.1992 (4 shells). —Debris of the Lez River, Castelnau, Mont- pellier, Hérault, France, M. Bodon leg. 3.12.1984 (many shells). —Debris of the Mosson River, Juvignac, Mont- pellier, Hérault, France, M. Bodon leg. 4.9.1985 (2 shells). Description Shell very small, planispiral, thin, pale whitish, waxen, transparent when fresh; sur- face of protoconch malleated; spire flat, con- sisting of 2.25-3 rapidly growing convex whorls; last whorl large, dilated, descending to some extent near aperture; umbilicus wide; aperture prosocline, roundish; peristome complete, slightly sinuous at upper margin, slightly thickened, reflected especially at lower and columellar margin (Figs. 213, 287; Paladilhe, 1967: 51, pl. 21, figs. 27-30; Lo- card, 1889: 329; 1893: 127; Germain, 1913: 323; 1931: 678; Boeters, 1974: fig. 8). Dimen- sions: height = 0.48-0.68 mm; diameter = 1.05-1.54 mm. Operculum slightly thickened, yellowish, paucispiral, thicker at centre of inner face, but without peg (Fig. 288; Boeters, 1974: 86). Body unpigmented; eye spots absent (Fig. 289; Boeters, 1974: 86). Male genitalia with prostate gland bulging slightly into pallial cavity; penis rather long, conical, having short, wide basal portion with corrugated sides and longer, gradually taper- ing, apical portion ending in very pointed tip; penial duct zig-zagging through right portion of penis to open at penis tip (Fig. 290; Boeters, 1974: 83, fig. 3). Female genitalia with distal seminal recep- tacle and a bursa copulatrix arising from distal renal oviduct; seminal receptacle, club-like, small but proportionally rather developed, arising from oviduct very close to point of ori- gin of duct of bursa copulatrix; bursa copula- trix large, sac-like, oval to square in outline, with very long, slender duct entering bursa at lateroposterior side; seminal groove running along ventral side of capsule gland (Figs. 292-294; Boeters, 1974: 83, fig. 4). Bursa copulatrix incorrectly depicted by Boeters (1974: fig. 4) as rather small with duct enter- ing on anterior side. Radula with central tooth trapezoidal with long lateral wings and basal tongue, its apical margin V-like with long, robust central denticle and 4-5 smaller denticles on both sides, in decreasing order of size; one basal cusp where each lateral wing arises from face of central tooth; lateral teeth rake-like, apically enlarged, their anterior margin with 9-10 den- ticles, central one longer, larger; first marginal teeth rake-shaped, with long lateral wing and elongated cutting edge with long row of 23-25 small denticles anteriorly; second marginal teeth scraper-shaped, with long slender lat- eral wing and roundish, spoon-like apex, its cutting edge carrying rather long row of 18-20 small denticles (Figs. 177-179). Stomach without posterior caecum; intes- tine with rather well-developed, U- or V-like bend on pallial wall (Figs. 291, 292; Boeters, 1974: 86, fig. 3). Osphradium oval or kidney-shaped; ctenid- ium consisting of 9-12 lamellae (Figs. 291, 292: Boeters, 1974: 86). Taxonomy Boeters (1974) assigned Valvata exilis to Horatia Bourguignat, 1887, which, at that time, was known to be characterized by: rather large bursa copulatrix and distal semi- nal receptacle (c.f., Radoman, 1966; Rado- man, 1973b). More recent anatomical study of Horatia revealed that the female genitalia not only have a bursa copulatrix and distal semi- nal receptacle, but also a proximal seminal re- ceptacle (Fig. 108; Radoman, 1983). Clearly, V. exilis does not belong to Horatia, and our anatomical study of specimens collected near the type locality brought to light new data which supports the introduction of a new genus. Only one of the genera with valvatiform shells described so far is close to Heraultia, that is, Arganiella Giusti & Pezzoli, 1980 (see above). Arganiella is characterized by a penis lacking lobes and by female genitalia with a large bursa copulatrix and only the distal sem- inal receptacle. Our decision to introduce Her- aultia is prompted by the fact that Arganiella is clearly distinguished by: penis dilated subapi- cally and with shorter apical portion; bursa 210 BODON, MANGANELLI & GIUSTI copulatrix having a rather short duct entering on the anterior side; central tooth of radula with two basal cusps on each side. Heraultia exilis was reported from Spain by Alonso (1975), Vidal-Abarca 4 Suarez (1985) and Bech (1990). All these reports are based on shell material and are therefore unreliable. ACKNOWLEDGEMENTS We thank Antonella Daviddi and Leonardo Gamberucci for technical assistance, Helen Ampt for revising the English, Reno Bernasconi (Münchenbuchsee, Switzerland), H. Girardi (Montfavet, France), Martin Haase (Vienna, Austria), Wim J. M. Maassen (Duive- drecht, Holland), Giuseppe Lucio Pesce (L'Aquila, Italy), Pavle Radoman (Beograd, Serbia), Rajko Slapnik (Ljubljana, Slovenia), Fabio Stoch (Trieste, Italy) and France Velkovrh (Ljubljana, Slovenia) for information about or loan of material from their respective collections, Simone Cianfanelli (Florence, Italy), Karl Edlinger (Vienna, Austria), Anita Eschner (Vienna, Austria), and Elena Gavetti (Turin, Italy) for bibliographical research. Research partly financed by MURST (In- vertebrate fauna of soil and underground wa- ters of the Apennine and insular Italy) and University of Siena (The non marine molluscs of the Italian Fauna) grants. LITERATURE CITED ALONSO, M. R., 1975, Moluscos terrestres y dul- ceacuicolas de la depresion de Granada (Es- pana) y sus alrededores. Cuadernos de Ciencias Biologicas Universidad de Granada, 4:125-157. ANGELOV, A., 1967, Horatia (Hauffenia) lucidula n. sp. ein neuer Vertreter der Molluskenfauna Bul- gariens. 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University of Michigan, Museum of Com- parative Zoology, Miscellaneous Publication, 6: 1-213. WILLMANN, R. & H. PIEPER, 1978, Gastropoda. In: J. ILLIES, [ed.], Limnofauna Europaea, Stuttgart, New York, Amsterdam: 118-134. WESTERLUND, C. A., 1898, Novum Specilegium Malacologicum. Annuaire du Musée Zoologique de l’Académie Impériale des Sciences de St.- Pétersbourg, 3: 155-183. ZIMMERMANN, S., 1930, Horatia erythropoma- tiana kerschneri n. subsp. Archiv fur Mollusken- kunde, 62: 233-234, pl. 13. Revised ms. accepted 29 July 2000 We. Ji MALACOLOGIA, 2001, 43(1-2): 217-222 IMMUNOLOGICAL DETECTION OF A CK2 ACTIVITY IN TAPES SEMIDECUSSATUS (MOLLUSCA, BIVALVIA) MANTLE L. Mercado', E. Itarte?, $. Marshall? & С. Arenas?* ABSTRACT Casein kinase 2 (CK2) is a highly conserved enzyme present in a wide range of eukaryotes, including yeasts, invertebrates and mammals. This kinase is involved in a number of key regu- latory cellular processes. We describe a CK2-like protein in mantle tissue of Tapes semidecus- satus, a highly active organ responsible for cell healing and regeneration in this clam species. After subcellular fractionation of crude homogenized mantles, the putative enzymatic activity was clearly enriched after two-step cromatography partial purification. Furthermore, using a poly- clonal antibody elicited against the known subunits of the mammalian enzyme, Western blot analysis confirmed the presence of three reactive polypeptides. Two of these perfectly matched the expected molecular weights of the catalytic a. (42 kDa) and a’ (38 kDa) subunits of the ref- erence enzyme. Considering our research interest is centered on the relationship between ma- rine organisms and their environment, our preliminary work on a CK2-like kinase in clams offers a potential model system to study protein phosphorylation in molluscs as an indicator of the ef- fect of environment factors over selected physiological responses. Key words: CK2, kinase activity, biochemistry mollusc, bivalve mollusc, clams, mantle, im- munodetection. INTRODUCTION The study of protein phosphorylation, one of the most relevant covalent modification of proteins, has demonstrated the existence of networks of protein kinases involved in the regulation of cellular processes related to me- tabolism, gene expression and cell growth. (Hunter & Cooper 1985; Krebs, 1994). One component of the network is CK2, also known as casein kinase 2, a pleiotropic and highly conserved serine/threonine kinase described both in the nucleus and the cytosol of both vertebrates (Tuteja & Tuteja, 1998; Chevet et al., 1999; Guo et al., 1999) and invertebrates (Hu 8 Rubins, 1991; Jaffe et al., 1997; Co- queret et al., 1998). The enzyme is also re- quired for viability as well as for cell cycle pro- gression (Pinna & Meggio, 1997; Guerra & Issinger, 1999). The prototype enzyme is composed of 4 subunits: 2 catalytic ones, a and a’ of 42-44 kDa and 38 kDa respectively, and two identical B regulatory subunits of 28 kDa each (Allende & Allende, 1995). In vivo CK2 exists as a tetrameric holoenzyme com- posed of either a,ß,, aa’B, or a’,B, (Chester et al., 1995; Gietz et al., 1995). A number of cytosolic and nuclear sub- strates of CK2 has been described in rat he- patocytes, a classical model, due to their re- generative capacity (Pinna, 1990; Issinger, 1993; Pancetti et al., 1996; Prowald et al., 1997; Roher et al., 1998; Miro et al., 1999). In this report we describe the activity of a CK2-like protein in a novel invertebrate sys- tem, the bivalve mollusc Tapes semidecussa- tus (Mediterranean clam). Moreover, the fact that the enzymatic activity is recognized by polyclonal antibodies against mammalian en- zyme subunits, strongly suggests a potential equivalent role of the enzyme in marine inver- tebrates. ‘Laboratorio de Fisiología Celular, Instituto de Biologia, Facultad de Ciencias Básicas y Matemáticas, Universidad Católica de Valparaíso, Chile. “Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad Autónoma de Barcelona, Spain. “Laboratorio de Genética y Inmunología Molecular, Instituto de Biología, Facultad de Ciencias Básicas y Matemáticas, Uni- versidad Católica de Valparaíso, Chile. “To whom correspondence: Laboratorio de Genética e Inmunología Molecular, Instituto de Biologia, Universidad Católica de Valparaíso, Av. Brasil 2950, Casilla 4059, Valpariso, Chile; garenas @ucv.cl 218 MERCADO ET AL. MATERIAL AND METHODS Tissue Preparation Twenty-five adult 7. semidecussatus speci- mens were collected from the delta of the Ebro river, Mediterranean Sea, Tarragona, Spain. The individuals were dissected in the laboratory, with a yield of 35 g of whole man- tles and kept under liquid nitrogen. The frozen material was homogenized in 100 ml of buffer A (50 mM Tris/HCl ph 7.4, 5% v/v glycerol, 1 mM DTT, 2 mM PMSF, 1 mg/ml aprotinin) using a steel mortar. The resulting ho- mogenate was submitted to serial centrifuga- tion steps, always recovering the supernatant: 800 x g (20 min), 19,000 x g (30 min) and 100,000 x g (75 min). The final soluble frac- tion, defined as S-100, contained 1.65 g of protein and was the source for enzyme en- richment. Partial Purification of CK2-like Protein To identify the enzyme activity, the whole S- 100 was loaded into a Whatman P11 phos- phocellulose column (3 x 1.5 cm) equilibrated with buffer A at 4°C, to a flow rate of 0.5 ml/min. The column was washed with 40 ml of the same buffer at the same flow rate and the adsorbed proteins were eluted through a saline gradient between 0.0 and 1.2 M NaCl in buffer A, into 45 individual 0.5 ml fractions. The mean 280 nm absorbance value was de- termined for each fraction to standarize the first CK2 enzymatic assay. Fractions display- ing enzymatic activity were dialized overnight against 1/1000 buffer A at 4°C. In order to enrich the enzymatic activity, a second step purification was done using a DEAE-Sepharose column (Sigma, DCL-6B- 100) packed into 2 ml and equilibrated with 20 ml of buffer A. The dialysed material was ap- plied to the column and eluted at a flow rate of 0.5 ml/min, through a saline gradient between 0.15 and 1.2 M Nacl in buffer A. Absorbance values at 280 nm as well as CK2 activity were determined. To further increase specific activ- ity of the resulting positive fractions, they were pooled, dialysed and concentrated into a 300 ul DEAE column, eluted at a one-step con- centration of 1.2 M NaCl in buffer A. Enzymatic Activity Assay CK2 activity was assayed measuring the in- corporation of °°P from ["32Р] GTP into B-ca- sein as substrate (Martos et al. 1985). The re- action mixture contained 10 ul B-casein (10 mg/ml), 15 ul of the test reagent and 5 ul of sample. The test reagent contained 2.085 pmol GT! “Ip, 8 mM EGTA, 0,2 M MgCl, and 5 mM ATP/GTP in buffer B (19 mM DTT, 10 mM EDTA, 500 mM B-glycerolphosphate) pH 7.0. The mixture was incubated for 15 min at 30°C, chilled and spotted onto p81 phospho- cellulose paper squares, fixed with ice-cold 10% TCA, and the radioactivity incorporated measured in a liquid scintillation counter. The activity of CK2 was expressed in mU/ml: one milliunit corresponds to the incorporation of 1 pmol of '***!P from GTP onto В-сазет (Plana et al., 1994). Protein Analysis and Immunodetection Protein concentration was determined by serial dilutions of samples assayed by the method of Bradford (1976) using bovine serum albumin as a standard. Polypeptide profiles of S-100, P11 and DEAE fractions were resolved by 10% SDS-PAGE (Laemmii, 1970) and stained with Coomassie brilliant blue. The enriched DEAE-peak fraction, resolved in a 10% SDS-PAGE, was electrotransferred onto Immobilon-P membranes (Towbin et al., 1979) for immunotesting using specific anti- bodies. Non-specific sites were blocked with 3.0% non-fat milk. The primary antibody, anti- rat liver CK2 antibody already purified by im- munoaffinity chromatography (Krehan et al., 1996), was a generous gift from Dr. P. Lorenz (Biochemische Zellphysiologie, Deutsches Krebsforschungszentrum DKFZ, Heidelberg, Germany). The mixture, a polyclonal antibody against o,a’, and В subunits was used at 1:4,000 dilution for one hour at room temper- ature with gentle agitation. The secondary an- tibody, goat anti-rabbit-IgG alkaline phos- phatase conjugated IgG, was used at 1:3,000 dilution for one hour at room temperature. The membrane was developed using the commer- cial enzyme substrate, BCIP (5-bromo-4- chloro-3-indolyl phosphate) and NBT (nitro- blue tetrazolium) (BIORAD). The rat liver enzyme was used as a positive control. RESULTS An S-100 extract from clam mantle tissue was the source to identify CK2 like activity after serial P11 and DEAE-cellulose column chromatography. Polypeptide enrichment after each step was confirmed using poly- CK2-LIKE ACTIVITY IN CLAMS 219 kDa 1 1s—> 62—> 51—> 38 — À 26—> 2 3 FIG. 1. 10% SDS-PAGE characterization of polypeptides resulting after sequential partial purification of a CK2-like enzyme from mantle crude extracts. Coomassie blue-stained profiles for identical protein concen- tration (30 ug) of 3 different purification steps: (1) S-100 fraction, (2) Post P-11 pool, (3) Post DEAE- Sepharose pool. 300 250 200 150 100 4 CK2 Activity (mU/ml) 50 12 16 20 24 Fraction 28 32 36 40 44 Number FIG. 2. Elution profile of S-100 from clam mantles on Phosphocellulose P 11 column chromatography CK2 referential activity in mU/ml across and the comparative protein concentration of each fraction as detected at 280 nm. acrylamide gel electrophoresis of equivalent protein concentration fractions (Fig. 1). Figure 2 shows the profiles of biochemical activity as well as protein concentration values after P 11 fractionation of S-100. CK2 activity was de- tected between fractions 22-32, with a peak in fraction 28, corresponding to an specific ac- tivity of 255.41 mU/ml. and to an elution NaCl concentration of 0.8 M. Figure 3 shows the elution profiles after the first DEAE column. The peak enzymatic activ- ity concentrates between fractions 12 to 18, matching with the highest protein concentra- tion. Western blotting assay of these samples revealed a 42 kDa polypeptide, which was recognized by anti-CK 2 a. It co-migrated at a similar molecular weight described for this subunit in mammals (Fig. 4A). 220 MERCADO ET AL. 300 250 200 150 CK2 Activity (mU/ml) 100 En — 50 Fraction Abs. 280 nm Number FIG. 3. Elution profile of post P 11 pooled fractions on a first DEAE-Sepharose column chromatography. CK2 referential activity in mU/ml and the comparative protein concentration of each fraction as detected at 280 nm. Fractions 12 to 15 (2 ml) were pooled, con- centrated to 300 ul and taken to a second DEAE column. The mixture was eluted as a unique fraction (post DEAE) at a single ionic strength of 1.2 M NaCl in buffer A. This frac- tion was probed with the anti rat liver CK2 polyclonal antibody. Figure 4B shows three discrete polypeptide bands, two of them corresponding to the rela- tive molecular weights of rat liver subunits © and с’ (42 and 38 kDa, respectively), and a third band at 60 kDa, which might very well be a species-specific В subunit, not equivalent in molecular weight to the rat liver counterpart, or simply anon specific reaction. We were not able to resolve either of these alternatives. DISCUSSION CK2 is an enzyme activity classically stud- ied in mammalian cells but not as yet in ma- rine organisms. Nontheless, the regulatory properties of eukaryotic CK2 enzymes are poorly understood (Guerra et al., 1999). The present work provides the basis for further studies on CK2- like activities in a new model system, marine bivalves, and the putative im- pact of protein phosphorylation in key biologi- cal processes, mainly those environmentally dependent. A number of features derived from our re- sults support the idea of the ubiquity of this enzyme. Firstly, the CK2 activity demon- strated in Tapes semidecussatus was at- tained using the same purification methods applied to mammals, and this could very well be considered a sign of biochemical and bio- physical conservation. Secondly, the fact that B-casein, the well- known specific substrate for the mammalian CK2 enzyme, displays a reasonable phos- phorylating activity using a non-purified puta- tive clam enzyme and GTP/ATP as donors, is another supportive argument for our proposal. Last, but not least, is the existing cross im- munoreactivity between the purified rat liver enzyme and the putative CK2 like clam coun- terpart. Considering that the polyclonal anti- rat CK2 were elicited against specific aminoacidic residues involved in subunit as- sociation residues 15-27 of the с subunit and 16-28 of the о’ subunit (Krehan et al., 1996), it becomes feasible that in the clam system a similar association exists that might represent CK2-LIKE ACTIVITY IN CLAMS 221 A 121371415 B ] > kDa ENE +60 o маи a ote Gm < 42 + 30 CK2 FIG. 4. A Western blotting analysis of fractions from DEAE (first column). В, Immunoblot of a post DEAE (sec- ond column) fraction (1) showing recognition of putative clam mantle CK2 subunits. Purified rat liver CK2 (2) as positive control. Equivalent protein concentration in both samples. homologs of mammalian enzymes. This is a known fact for a number of different species, such as frog (Jedlicki et al., 1992), chicken (Maridor et al., 1991), and human (Lozeman et al., 1990) when compared to the rat en- zyme. Thus, the similarity between the a: subunits of mollusc and mammals would indicate that the in course of evolution, the catalytic sub- units could have been preferentially con- served and not necessarily the regulatory В. The relevancy of this point is that the catalytic subunits of the enzyme are the ones acting upon the target substrates. CK2 В subunit was not detected as expect in our system. One possibility is that amino acid residues 171-186, the region from the mammalian subunit of CK2 specifically recog- nized by antibodies (Krehan et al., 1996), are not present in putative subunits of the mollusc kinase. On the other hand, the unspecific de- tection of a 60 kDa polypeptide instead of the expected 28 kDa reported size for the mam- malian subunit (Meggio et al., 1992), might also mean that in mollusc it is in a dimeric or larger form. In fact, the literature reports a slightly larger В subunits in the yeast $. cere- viseae (Bidwai et al., 1994). ACKNOWLEDGEMENTS L. 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TOWBIN, H., T. STAEHELIN & J. GORDON, 1979. Electrophoretic transfer of proteins from poly- acrylamide gels to nitrocellulose sheets: proce- dure and some applications. Proceedings of the National Academy of Sciences USA, 76: 4350- 4354. TUTEJA, В. & N. TUTEJA, 1998, Nucleolin: а mul- tifunctional major nucleolar phosphoprotein. Crit- ical Reviews in Biochemistry and Molecular Biol- ogy, 33: 407-436. Revised ms. accepted 10 November 2000 MALACOLOGIA, 2001, 43(1-2): 223-236 INFERENCES ON THE PHYLOGENETIC RELATIONSHIPS OF SUCCINEIDAE (MOLLUSCA, PULMONATA) BASED ON 18S rRNA GENE Ana Virginia Calogeras Dutra- Clarke”, Charlene Williams”, Rebecca Dickstein’, Norbert Kaufer® 8 James В. Spotila!* ABSTRACT The systematic position of Succineidae and its relationship with Athoracophoridae and other pulmonate mollusks is controversial. In this paper, we attempted to study their relationships in the light of the 18S rRNA gene. We determined complete sequences from the 18S rRNA gene for Omalonyx matheroni (Potiez & Michaud, 1835), a closely related species to Succinea putris, the type-species of the family Succineidae (Tillier, 1981), and Athoracophorus bitentaculatus (Quoy & Gaimard, 1832), the type-species of the family Athoracophoridae. All complete se- quences of closely related pulmonate molluscs available from GenBank were included in the phylogenetic analysis in an attempt to find the positions of these taxa in the pulmonate tree. Comparative sequence analysis and base stacking energy methods were used to predict the secondary structure of the 18S rRNA gene. The secondary structure was used to resolve am- biguous nucleotide positions for our alignment. Stems E10-1, E23-1, 43 and 49 of the 18S rRNA gene were highly variable and the suggested secondary structure was presented for 21 mol- lusks. We used maximum parsimony and maximum likelihood methods for assessment of the re- lationships of the Succineidae. The 18S rRNA data indicate Succineidae and Athoracophoridae taxa form a monophyletic clade, the Elasmognatha. The Elasmognatha is positioned among the stylommatophoran molluscs. The order Stylommatophora is monophyletic. The subclass Pul- monata is not supported. Siphonariids are an early diverging lineage to all taxa or positioned as a sister taxon to the pulmonates. Furthermore this study indicated that siphonariids are not part of or even closely related to the main lineage of basommatophorans represented here by Planor- bidae and Lymnaeidae. Key words: 18S rRNA gene; secondary structure; Succineidae; Athoracophoridae. INTRODUCTION The mollusk subclass Pulmonata, or the lung breathing snails, includes the vast major- ity of terrestrial, numerous freshwater, and a few marine sublittoral mollusks. Their main ra- diation occurred among the representatives of the land snails order Stylommatophora, in which 71 to 92 families are reported (Ember- ton et al., 1990), and the number of species are estimated to be between 30,000 to 35,000 (Solem, 1984). The fossil record dates from the mid-Paleozoic (Solem, 1979; Solem & Yochelson, 1979). The Pulmonata monophyly was strongly supported with partial se- quences of the 28S ribosomal RNA by Rosen- berg et al. (1994), who also pointed out the monophyly of the order Stylommatophora. Succineidae are distributed worldwide, with their origin considered as Laurasian (Tillier, 1989) or Gondwanaland (Nordsieck, 1986). Athoracophoridae are considered to be a sis- ter group to Succineidae, according to mor- phological data (Nordsieck, 1986) and partial 28S rRNA gene sequence analysis (Tillier et al., 1992), although their close relationships have been rejected in other studies (Solem, 1978; Tillier, 1984, 1989; Schileyko, 1978). The Succineidae + Athoracophoridae clade was named as Elasmognatha, because, among pulmonate snails, the included taxa share a unique type of jaw called the elas- mognatha jaws. The Succineidae are a sty- lommatophoran family of land snails that oc- cupy a key position in the elucidation of the steps of pulmonate evolution. Tillier et al. "Department of Environmental Sciences, Engineering and Policy, Drexel University, 32nd & Chestnut Streets, Philadelphia, Pennsylvania 19104, U.S.A.; dutra-clarke O worldnet.att.net Department of Biochemistry and Molecular Biology, Jefferson Institute of Molecular Medicine, Jefferson Medical School, Thomas Jefferson University, 233 So. 10th Street, Philadelphia, Pennsylvania 19107, U.S.A. ‘Institut für Genetik, Biozentrum Technische Universität Braunschweig, Spielmannstrasse 7, D-38106 Braunschweig, Ger- many. “To whom correspondence should be directed; spotiljr@ drexel.edu 224 DUTRA-CLARKE ET AL. (1992) reestablished the taxon Elasmognatha by showing the monophyly between Succinei- dae and Athoracophoridae based on partial sequences from 28S rRNA gene. The Elas- mognatha seemed to retain many symple- siomorphic characters, and thus their classifi- cation based on morphology has been deeply affected by them (Dutra-Clarke, 1996). The systematic position of Succineidae within the Pulmonata has been a crucial problem in un- derstanding pulmonate evolution. Tillier et al. (1992) showed Succineidae and Athora- cophoridae related to Opistobranchia, and they stated: “. . . the position of this taxon, sister-group of the order Pulmonata or inside the Stylommatophora, varies in relation to minor changes in alignment. . . .”. We ap- proached the problem of their relationships by means of a detailed analysis, including sec- ondary structure information, of complete se- quences of the 18S rRNA gene. The type- species Athoracophorus bitentaculatus was selected to represent Athoracophoridae. The Succiniedae is represented by the semi-slug Omalonyx matheroni. Omalonyx is related to Succinea, based on combined analysis of morphological data and partial sequence data from 18S rRNA gene (Dutra-Clarke, 1996) and on the hypothesis that Omalonyx evolved from Succinea (Tillier, 1981). The small subunit rRNA or 18S rRNA gene is the most studied rRNA gene. In relation to the land mollusk order Stylommatophora, up to the present time, 18S rRNA sequences have been published for Achatinidae, Clausili- idae, Helicidae, and one Succineidae (Win- nepennickx et al., 1992, 1996, 1998). We are adding two more in this paper, that of a Suc- cineidae and of the type-species of Athora- cophoridae. The 18S rRNA gene remains ho- mogeneous as a result of concerted evolution (Hillis & Dixon, 1991). This characteristic plus the diversity of domains from conserved to highly variable make the 18S rDNA full-length sequence a key molecule to approach rela- tionships from species to class level. Full-length 18S rRNA gene has been used to elucidate relationships within the mollusks (Adamkewicz et al. 1997; Kenchington et al., 1994; Winnepennickx et al., 1992, 1994, 1995, 1996, 1998a, b; Bargues & Mas-Coma, 1997) and closely related protostomates. Adam- kewicz et al. (1997) studied relationships of subclasses within Bivalvia. Kenchington et al. (1994) analyzed family level relationships; Winnepennickx et al. (1992, 1994, 1995) used 18S rRNA gene to establish the monophyly of the phylum Mollusca and (1996) the relation- ships among classes of the phylum Mollusca. Winnepennickx et al. (1998b) found the 18S rDNA could also resolve for “rapid” recent cladogenetic events, such as divergences between species of Littorinidae. Moreover, Bargues & Mas-Coma (1997) discriminate species of the family Lymnaeidae based on the sequence and characters of the secondary structure of 18S rRNA gene. The estimated ratio of transition to trans- version for our full length data-set was of 1:1. Therefore, substitutional saturation of sites seems not to be a problem. For a site that is at or near saturation, this ratio would be of 0.5 (Aboudheif et al., 1998). Even when we elim- inate the highly variable sites of the molecule (bulges, loops and unpaired nucleotides on the secondary structure), this ratio was of 1:0.93, and a similar phylogeny was obtained. The purposes of this study were (1) to de- termine if 18S rRNA gene sequence data sup- port the hypothesis that the families Succinei- dae and Athoracophoridae belong to a monophyletic clade; (2) to test whether the Succineidae and Elasmognatha belong to the main pulmonate radiation, that of the stylom- matophorans, or if they are sister-group to the stylommatophorans; (3) to test the relation- ship of Succineidae and Achatinidae, consid- ered by Tillier (1989), based on morphology, to be more closely related than is Succineidae to Athoracophoridae; (4) to present a compar- ative analysis of the secondary structure of highly variable subdomains of the 18S rRNA gene for mollusks showing its importance for the phylogenetic inferences. MATERIALS & METHODS Specimens of the semi-slug Omalonyx matheroni were freshly collected from Brazil (vouchers deposited at the Academy of Nat- ural Sciences — ANSP A18026-A18038; 18S rRNA sequence data at GenBank, accession number AF047199) and Athoracophorus bi- tentaculatus, the type-species of Athoraco- phoridae, were museum preserved speci- mens from New Zealand (ANSP A7965b, d; 18S rRNA sequence data at GenBank, ac- cession number AF047198). Closely related outgroup to pulmonates is represented by a Neogastropoda; another more distant related outgroup is represented by a Vetigastropoda. Published complete 18S rRNA gene se- quence data are the vetigastropod Mono- PHYLOGENETIC STUDY OF THE FAMILY SUCCINEIDAE 225 donta labio (X94271); the neogastropod Thais clavigera (X91979); and the euthyneurans Aplysia sp. (X94268), Lymnaea_ stagnalis (Z73984), Fossaria truncatula (Z73985), Ba- kerilymnaea cubensis (Z83831), Radix pere- gra (Z73981), Stagnicola palustris (Z73983), Lymnaea glabra (Z73982), L. auricularia (Z73980), Biomphalaria glabrata (U65224), B. alexandrina (U65225), Siphonaria alge- sirae (X91973), Onchidella celtica (X70211), Limicolaria kambeul (X66374, Achatinidae), Helix aspersa (X91976, Helicidae), Oxyloma sp. (X94276), Balea biplicata (X94278), and Laevicaulis alte (X94270). Taxonomic groups resolved in our tree follow names proposed by Ponder & Lindberg (1997). DNA Extraction Foot muscle and liver tissue were taken from dissected alcohol preserved specimens (A. bitentaculatus) or only the foot muscle was taken from fresh-frozen snails (O. matheroni). DNA was obtained from at least two individuals of each species. Total DNA was extracted by an adaptation of the CTAB (cetyl- trimethylammonium bromide) method as de- scribed for plant DNA using CTAB as in Cur- rent Protocols in Molecular Biology (Ausubel, 1994) and in Spolsky et al. (1996). PCR Reactions The PCR reactions were performed with GeneAmp (Parkin-Elmer Corp) on a Coy Tem- pcycler. The PCR primers are listed in Table 1. Reactions were performed in a total volume of 50 ul using the ampli-Taq DNA polymerase kit (200 uM of each dNTP; 1 uM of primers and 2.0 U to 2.5 U of ampli-Taq DNA polymerase). The concentration of the template DNA varied between 50 ng and 100 ng. The standard PCR amplification profile consisted of an initial de- naturation step of 95°C for 2 min, followed by 25 to 30 cycles of 94°C for 1 min, 48-51*C for 1 min, 72°C for 1 min and 30 sec, followed by 10 min at 72°C for final extension. The purifi- cation of the PCR product was done with either the Magic Clean kit from Promega or with the Qiagen PCR purification kit from Qiagen Inc. The purified PCR product was then prepared for direct sequencing. Sequencing Both DNA strands of the 18S rRNA gene were sequenced. Automated sequencing used 3.2 pmol of each PCR primer to prime the cycle sequencing reactions in both the sense and antisense directions. The auto- mated cycle sequencing was performed in a Model 373A or Model 377 DNA sequencing system (Applied Biosystems Inc., Foster City, California), at the Department of Biochemistry of Thomas Jefferson University, Philadelphia, Pennsylvania. The sequences were read and entered into the multiple-sequence editor ESEE (Cabot & Beckenbach, 1989). The primers were complementary to con- served regions of the eukaryotic 18S riboso- mal gene and are listed in Table 1. Secondary Structure and Sequence Alignment Compilations of rRNA small subunit struc- tures (Neefs et al., 1993; Van de Peer et al., 1996) and the predicted secondary structure for the pulmonate snail L. kambeul (Win- nepennickx et al., 1992) were used as models to obtain the secondary structures here pro- posed. We followed the method described by Kjer et al. (1994) and Kjer (1995) to manually predict secondary structure and to write it on the aligned sequence. This comparative se- quence analysis is considered by Guttell et al. (1994) as a very refined test of homology and is based on the concept of positional covari- ance. Woese et al. (1990) showed that com- parative sequence analysis has been in- volved in the derivation of higher-order structures for a variety of different RNA mole- cules. We also applied the RNAdraw program (Matzura & Wennborg 1996), which uses min- imum energy structure prediction algorithm to fold the RNA to obtain diagrams of the sec- ondary structure. If used exclusively, this pro- gram can offer poor resolution. Optimal struc- tures could only be obtained when constraints for nucleotide pairings were previously de- fined according to the comparative analysis of aligned sequences. The diagrams of the pre- dicted secondary structure were used to ei- ther corroborate or decide for difficult homolo- gous nucleotide positions in our alignment. We followed the stems numbering as in Neefs et al. (1993). However, stems 20 and 21, as seen in Neeffs et al. (1993), compilations were not drawn for the pulmonate secondary structure model of L. kambeul (Winnepen- ninckx, 1992). We numbered the stems as Neefs et al. (1993) but omitted stems 20 and 21 following the structure presented by Win- nepenninkx et al. (1992), because they are 226 DUTRA-CLARKE ET AL. TABLE 1. List of primers ID Sequence Publication Position* #2 CAACCTGGTTGATCCTGCCCAGT Rice (1990) 0-22 NS1 GTAGTCATATGCTTGTCTC White et al. (1990) 19-38 A AGGGYTCGAYYCCGGAGA Winnepenninckx et al. (1994) 391-409 $23 GGAGCATGAGAAACGGCTAC This paper 431-410 NS3 GCAAGTCTGGTGCCAGCAGCC White et al. (1990) 572-593 NS2 GGCTGCTGGCACCAGACTTGG White et al. (1990) 593-572 B CGCGGTAATTCCAGCTCCA Whinnepenninckx et al. (1994) 592-611 C TTGGYRAATGCTTTCGC Whinnepenninckx et al. (1994) 987-970 D TTAATCAAGAACGAAAGT Whinnepenninckx et al. (1994) 998-1016 G CCGTCAATTCCTTTAAGTTT Whinnepenninckx et al. (1994) 1185-1165 NS4 CTTCCTCAATTCCTTTAAG White et al. (1990) 1188-1168 = AATTTGACTCAACACGGG Whinnepenninckx et al. (1994) 1216-1234 NS7 GAGGCAATAACAGGTCTGTGATGC White et al. (1990) 1449-1472 F GGGCATCACAGACCTGTTA Whinnepenninckx et al. (1994) 1474-1454 | TTTGTACACACCGCCCGTCG Whinnepenninckx et al. (1994) 1663-1680 GACGGGCGGTGTAC Whinnepenninckx et al. (1994) 1679-1663 ITS1 GGAAGTAAAAGTCGTAACAAGG This paper 1782-1807 NS8 TCCGCAGGTTCACCTACGGA White et al. (1990) 1829-1809 #1 CTGATCCTTCTGCAGGTTCACCTAC Rice (1990) 1836-1811 "Position as compared to the published sequence of L. kambeul (Whinnepenninckx et al., 1994) not well formed for the molluscs studied here. The highly variable subdomains of the do- mains V2, V4, V7 and V9 in the 18S rRNA gene for the 21 mollusks studied here were represented by the stems E10-1; E23-1, 43 and 49. Non-canonical base pairings G:U were common, but the non-canonical G:A were not allowed. Once the alignment for the highly variable regions V2, V4, V7 and V9 were ready, and with the guidance of the sec- ondary structure, the rest of the 18S rRNA gene could be easily aligned. All diagrams were redrawn manually. We aligned the new complete 18S rRNA gene sequences with complete published se- quences for the pulmonates from GenBank. Very divergent domains of the 18S rRNA gene were difficult to align. However, with the sec- ondary structure predicted for each taxon, ho- mologous nucleotide positions were defined and a careful manual alignment resulted in a confident data set and allowed us to include the entire gene sequence into our analysis. ESEE multiple sequence editor software (Cabot & Beckembach, 1989) was used for the alignment. Gaps were inserted to account for length differences between the se- quences. Phylogenetic Analysis The two new complete 18S rRNA se- quences were manually aligned with pub- lished sequences. The sequences were for- matted for phylogenetic analysis using the software EAT (Cabot, 1993). The ratio of tran- sition to transversion was empirically deter- mined by performing likelihood analyses on all taxa, testing a range of ratios varying from 0.4 to 1.5. The ratio which yielded the higher log likelihood for the data was than selected. Phylogenetic analysis were performed using parsimony (PAUP 4.0d57; Swofford, 1998) and maximum-likelihood (from PHYLIP; Felsenstein, 1993). Gaps were treated as missing data. Unweighted parsimony trees were found by conducting a heuristic search. The tree nodes were tested for reliability by bootstrapping (500 replicates on the full length sequencing data and 100 on the data-set ex- cluded of highly variable sites). Majority rule consensus trees were generated from the bootstrapped trees. We selected a more closely related species to the Euthyneura (the neogastropod Thais clavigera) and a more distant taxon (the vetigastropod Monodonta labio) based on gastropod relationships shown by Ponder & Lindberg (1997) and Win- nepenninckx (1998a). We applied two different tree-making meth- ods to 18S rRNA gene sequences, the maxi- mum likelihood (dnaml from PHYLIP, Felsen- stein, 1993) and parsimony (PAUP 4.0d57; Swofford, 1998). They differ in their assump- tions on the evolutionary process, and could well constitute a test on the reliability of our data to generate a robust phylogeny. PHYLOGENETIC STUDY OF THE FAMILY SUCCINEIDAE 227 RESULTS Alignment and Secondary Structure The alignment of 21 taxa relied on the pro- posed secondary structure yielded 1926 com- parative sites, of which 176 were parsimony- informative. Secondary structure was predicted for the stems E10-1, E23-1 43 and 49 (Figs. 1-4) be- cause of the variation found in nucleotide composition among the 21 aligned se- quences. The predicted secondary structure allows phylogenetic resolution for family and order level of relationships among the studied species representing families. Variability in stem E10-1 was detected among succineid genera and between succineids and other pulmonates (Dutra-Clarke, 1996). GNRA tetra-loops were found in stem E10- 1 and 49 (R stands for Purine and N for any nucleotide). These loops are considered to be a common RNA structural motif and an un- usual stable hairpin loop structure (Woese et al., 1990; Jucker & Pardi, 1995). Another tetra-loop sequence UCGC was found in the stem E10-1 for Athoracophoridae and Suc- cineidae; in the stem E23-1 for Achatinidae, Athoracophoridae, Helicidae, Succineidae, Lymnaeidae, Onchididae, and Siphonariidae and in stem 49 for Onchididae. Phylogenetic Analysis The phylogenetic position of Succineidae and Athoracophoridae (Elasmognatha) rela- tive to others mollusks was inferred using maximum parsimony (Figs. 5, 7) and maxi- mum likelihood (Fig. 6) analyses. Tillier et al. (1992) consensus tree shows the Elasmog- natha in sister group position to all other pul- monates. Parsimony generated a consensus tree: length of 739 steps, consistency index (Cl) of 79; homoplasy index (HI) of 0.21, re- tention index (RI) of 0.74, and rescaled con- sistency index (RC) of 0.58 (fig. 5). The maxi- mum likelihood tree has a log of -6803.85 (Fig. 6). Contradicting the overwhelming morpho- logical evidence presented by Dutra-Clarke (1996) for the status of the family Succineidae as monophyletic, the molecular data support the hypothesis that Succineidae is para- phyletic due to shared sequence homologies between Athoracophorus and Omalonyx, di- verging from the published sequences of the succineid taxon Oxyloma sp. Therefore, the taxa Elasmognatha (reestablished by Tillier et al., 1992) is here defended, and it is supported by 99% to 100% of the bootstrap value. There- fore, there is no indication of close relation- ships between Succineidae and Achatinidae, as supposed Tillier (1989). The subclass Pul- monata was not supported due to the poorly resolved relationships between the systelom- matophorans and siphonariids with the opisto- branch Aplysia, the relationships of which are unstable in the parsimony or maximum likeli- hood trees. There is an unresolved polytomy consisting of four clades — (1) “higher” ba- sommatophorans + stylommatophorans, (2) systelommatophorans, (3) Siphonaria, and (4) Aplysia sp. The taxa Euthyneura formed a ro- bust clade among gastropods, which is con- firmed with 100% confidence. Furthermore, our analysis indicates siphonariides are nei- ther part of the basommatophorans clade here represented by Planorbidae and Lymnaeidae, nor are they closely related to them. Among the “higher” basommatophorans, the family Lymnaeidae and the family Planorbidae are monophyletic and supported with 100% confi- dence. Among Lymnaeidae, there are two lin- eages, but the only clade well supported by 99% to 100% bootstrap is formed by Lymnaea glabra as sister taxon to L. stagnalis and Stag- nicola palustris. This clade was also well sup- ported in the parsimony tree of Bargues & Mas-Coma (1997), and a name could be ap- plied for this node. The family Planorbidae is well supported with 100% bootstrap in both trees. For the data-set excluded of highly vari- able sites, parsimony generated a consensus tree: length of 690 steps, consistency index (Cl) of 0.77; homoplasy index (HI) of 0.23, re- tention index (RI) of 0.71 and rescaled consis- tency index (RC) of 0.55 (Fig. 7). There is a tri- cotomy, due to collapse in nodes representing species divergency among lymnaeids. These missing nodes were poorly supported in the trees generated by using full-length gene se- quence (Figs. 5, 6). The only robust node uni- fying the lymnaeid species Stagnicola palus- tris, Lymnaea glabra, and L. stagnalis is present and well supported by 70 of bootstrap value. DISCUSSION Boore & Brown (1994) appropriately com- ment: “a sequence alignment is fundamen- tally an hypothesis of homology at each of the aligned positions; changing the alignment of nucleotides can generate very different evolu- tionary trees.” Once we obtained the se- DUTRA-CLARKE ET AL. ou ou > o > o >—o >-u oo oo o-o > о v—u 0-4 >-o ou o-u 2—< о uu — © =) м = 2 2-u 2-4 oo o-o o-o 2-0 0-0 ou oro o—U 3—@ oO “о бо _o a o > o-> vou v—-uu® o-u?>o oo ou >-< o-< >o 4-> <— 2 Я <-> Я «—5> Я <-5 “ло -о “о 0 Vo-u o-u oe о-8 «<-> => <-> < o es о < < < < < < < < < < o 2 o 2 o = =) 25 2 > = o > o > о © > о | о > < (0) < o < о 2-0 2-0 2-9 2—< 2—< 2—< <-> «—> <-> a 2—<« À a >—< À o 2—< . < ®\5-0 a 2 a 2 o 2— o->> <-> x e я <-> 2 4—> 2 q 2) 0-40 “``. TC o..o 0 —_u uu u-0 o-> <-> <-> < о < © < о < < < < < < < o < © o о 9 о 5 o 5 > 2 5 = о v Jo 2 =. 5 =, > о 2 o о a © o < о-о 2-0 2-0 > —< D —< 2-4 = <-> <-> <-> —_—< 3 2 2 —4« = 2—< 5 #82 2 a qa” 58 PA | = 2-< 9 bh в E съ < o» о < о 5 9 Die >-< > © O0 o—-0. 2-0 oo o-o au оо =u = 2° ne 2—0 —-o GES ou ou A >-u ees o-o -o 0-03 en o 5 D2—<0° <—- >00 < 5° я <-—5 <- : Se Boo во oo E) ee < © = 2 < < > < A y o o > > 2 > o > > > o = [С] < о < о >-0 2-9 >-« >-< <=> ; <-2 & 2—< $ Dar gs” dise 5 2 A т nm © « © < IA u—o uo o—u 0-3 oo —0 ou o—0 2 o w—2 2 —0 = _ © o ом © o.—o 5-6 en uu o-> _ 2-0 ао ee oie — Oo — 220 826 — uw =) о-955 Voy? 5—« >-au® < o <-> 8 «<-> & <-> o-u ou o—v o-u <-> a < о > о < < < < < о « о bd ae > о о 5 в 5 =) © = © = < o o >-o >-u >-< >-< <—2 <-> 2 D>-a $ 2— = 2 < pro RAS < о ™ o . о 276 255 27 218 275 »-0 2—< «<-> 2—<« 5-4. в 223 2-26 > —< <-> > —< FIG. 1. A, B. Secondary structure representations for stem E10-1 in the 18S rRNA gene for 21 taxa. ondary structure. As Kjer (1995) pointed out, quences for this study one of the most careful the conservation of the rRNA secondary structure exceeds that of its nucleotides and and time consuming steps, was the align- ment. Secondary structure is suggested for should be used for the assignment of homol- stems E10-1, E23-1, 43 and 49, because ogous positions for phylogenetic studies. Many authors exclude highly variable regions those regions were ambiguous to align with- out the knowledge and guidance of their sec- 229 664 3CCGU уси GCCCGUC GGG GGUAA _ 645 U A с GU GUCC UUGUU 11 EDER ee UG G cUG,GGcGcG 659 PHYLOGENETIC STUDY OF THE FAMILY SUCCINEIDAE 5 GGGU GU UCGC EIS IA / 3 CCUG MON 63 (A) G с с G cc U с U G U G | с UCUG CCCGUCA GGCCG U с G 1 | G U U U cc A | U с | 6 658 U 1 А с с GCCG 641 701 679 678 ео ео о оо > o o-oo > > ou о-о o-u wien LS} о-о oS ou е-о Se oso. OS) оо >) = >-«c a Seo E) o =0 9 Bry—o o > OU—2 o-> > OF o-u оо SO uo oo DES 2 =o) o-oo ES u-o 0 o-v = ore be usa ee 0-5“ Sa GS-30 ¿231 <—5<“« 5 < < < о о < 5 o > > o o SI et U—20 DY me uw vn 2—0 a 9-0, = AS HUA OS a O=— 0 < "uo AS na bo om nm bm” ou ео 5 о > o ou ou vu-o o-v ou е-о u—o 2 e, o-u ou ou v-u> o-u> o-9 = > 0-0 2-0 2—, UE? a Gee $ 5-<«о o-u RC roo 0-0 U=0 0-3 U—o Vo o-u a oe: = = -U = BES => DENT ces o = Cn VU == = o-> <—2< 4-3 < < u—ow = o E <—2 o > > 5 o o o o v=>0 o->0 uU=> 2-1 9 23-0 © = UN B=-0,R 2 0-0,X <3. 0-0, 8 ‘u—o n u—o 2 uo о в E om oO i à FIG. 2. A, B. Secondary structure representations for stem E23-1 in the 18S rRNA gene for 21 taxa. 230 (A) (B) FIG. 3. A, B. Secondary structure representations for stem 43 in the 18S rRNA gene for 21 taxa. MON 1349 G cu S'CCU CUAA 1363 AGUU DUTRA-CLARKE ET AL. cGcc GACAG © 3'GGA GAUU UCAA GCG < E UGUUG \ ACA—U 1415 > G=C GC UA GU 6-0 ТНА cu 1 93, zu ABT S'CCU CUAA AGUU CGCCGAU Su A ANA с 3GGA GAUU UCAA GCGGCUG A cu UC сви 1405 APL 1354 1368 A AU e ccu S'CCU UUAA AGUU CGCCGGU у ТЕР TEED badd Е ЕЕ c GGA GAUU UCAAGCGGCCG G à cu с UguA 1406 ATH 1358 1362 / A AU сс Cc S'CCUUUAA AGUUGCGCCGGU N III dil Peel с Y GGA GAUU UCAACGCGGCCG, с \ cu 1412 QUA BAK 1391 1405 Y A AU / S'CCU UUAA AGUUCGCC UCGGUGGU CC A CA OA A TO OA 3'GGA GAUU UCAA GCGGAGCCGUUA \ A с су C 144 BAL 3351 1365 BER AU / cfc S'CCU UUAA AGUU CGCCGGU U Ir IATA U YGGA GAUU UCAA GCGGCCG . G \ с с GuA 1403 LYA 1381 1395 Y A AU U Y 5S'CCU UUAA АСИ CGCC UCGGUG GU byt UE ott ВВГ 3'GGA GAUU UCA GCGGAGCCGUUA \ cu c u с 1438 А LYG 1380 1394 RER AU 2 S'CCU UUAA AGU” ССС U Cee рр 6 IIA GGA GAUU UCA GCGGA \ cu си U 1437 A LYS 1380 1394 / A AU + S'CCU UUAA AGU © GGA GAUU UCA 1437 OMA 1351 7 А 5'CCU UU AA AAA A! 3} GGA GAUU 1403 ONC 1375 Y A A 5'CCU UUAA LA PE BEIN I GGA GAUU 1427 cu cu су U AU 4 1389 Y cc ЧА вии CGCCGGUS Uy UCAA GCGGCCG с с GU cécc UCGGUGGUY AA A GCGG AGCCGUUA, с G с BIA 1375 1389 / А AU / uc Cy SCCU UUAA AGUU CGCCGGU с Pn CE AS A EN G 3 GGA GAUU UCAA GCGGCCG A ` cu с CGu 1427 BIG 1376 1380 / А AU < ис SCCU UUAA AGUUCGCCGGU LOE EC vant G Y GGA GAUU UCAA GCGGCCG A N U C cu 1428 FOS 1374 1388 EM AU / S'CCU UUAA AGUUCGCC UCGGUGGU 9 CAC D PA EA DER re OR EE GGA GAUU UCA A GCGGAGCCGUUAY \ cu cu С 1431 HEL 1368 1372 7 A AU / с сс 5S'CCUUUAA AGUU CGCCGGU U Me CE en с Y GGA GAUU_ UCAA GCGGCCG 6 A 1419 U LAE 1353 1367 Y A AU Y U S'CCU UUAA AGUUCGCCGGU UC PC Ve ze ET! YGGA GAUU UCA A GCGGCCG Аб | > cu с 7U 1403 UM 1371 1385 / A AU / с S'CCU UUAA AGUU CGCCGGU UC LTE Тре TY Y GGA GAUU UCAA GCGGCCG Аб с N u Cc cu 1422 OXY 1347 1351 А AU 4 с S'CCU UUAA AGUU CGCCGGUS Ca TA ae al с 3'ббА GAUU UCAA GCGGCCG 6 ` cu с Cola 1399 u RAD 1 36 1390 7 À AU US SCOCUUUAA АСИ © cécc UCGGUGGUY A TACA ET ES CUIR | A TOR aA Ve GGA GAUU UCA GCGGAGCCGUUA y \ сие с 1443 А sıp 1373 1387 SS AU 1 с SCCU UUAA AGUU CGCCGGUS Cy Pe ATA IA с 3'ббА GAUU UCAA GCGGCCG \ CU с UA 1424 STA 1382 ase / AU SOCUUUAA АСИ © cocc UCGGUGGUY GGA GAUU ven N cu 1439 GCGG AGCCGUUA y U U с PHYLOGENETIC STUDY OF THE FAMILY SUCCINEIDAE (A) von 1675 SE GA 1719 / А GA GA CAGA AG / S'UACU CC UUGAAUGGUU Yau GACC UC UGG UCCGUCUGCCGCUU OT TOT OS О III YA UGA GG GAUUUACUAG UCA CUGG An GGC, GGGCAGGCGGUGGG A А AGDE ANTAS 1785 Pi THAIS 1663 1677 5, 1707 / А GA / y 6A GA cc р S'UACU CC UUGAAUGGUU AGU 6660 “ue UUGGUCU cGGc GCCUUU д A A Tee A: DANSE AA AUUVSCUAE UCA CUCG AG AGCUGG GCCG CGG 666, AG с a G UU с 174 APL 156 1600 G mo GA с бА GaGa’ U / U АСУ uc UUGAGCGGUU AGU GGGC UC UUGGUC CGGUCUGGCGU ¢ AAA A dl J'AUGA AG GGUUCGCUAGUCA CUCG a AGCCGG GCCAGGCCGCG 6 SATA c AG САБ шос E 1777 oA ATH 1669 1683 G 1713 ИА ВА / С СА CIEGA 5'UACU UC UUGAGCGGUUAGU GGGC UC YUGGUC UCGGUCUGGCAC € IA A ae EL CHE A TE JAUGA AG GGUUCGCUAGUCA CUCG AG AGCCGGGGCCAGGCCGUG A AG SATA с AG A UU с 1781 BAK 1709 thee) 6 1753 / A GA / с бА CGA U / G S'UACU UC UUGAGCGGUUAGU GGGC UC CUGGUC CGGUCUGGCGU с A ae TT RENE A FAUGA, AG AE SUUCGCUAS UCA CUCG AG GGCCGG GCCAGECCECG, \ C AG AS 1820 pa (B) 1676 18 GA c GOA 6 SUACU “uc UUGAGCGGUU AGU GGGC UC UUGGUU C6GUCUGGCGU с ЕЕ НО PEPE EE НЕЕЕЕТИНЕНИНЕ О AG ОВО UCA CUCG AG AGCCGG GCCAGACCGUA, A Age) СА 6 UC € im A UM A 16m 1695 175 EN EN A ea / 6 S'UACU UC UUGAGCGGUU AGU 6666 UC UUGGUC CS GUCUGGUAC с ПО О СО СООО EC CT TN TN CCR Bauch AG GGUUCGCUAG UCA CUCGAG AGCCGG CIA A А А с Аб AA ae uc с m LYA 1 us A GA С GA С GA SUACU uc VUGAGCEGUL AGU GGGC UC UUGGUC Vegcu cugccau® с НЕЕ НО ТНЕНЕН EU TEEPE EE ТН PPT YAUGA AG GGUUCGCUAG UCA CUCG AG AGCCGG GCCA 66СС6С6 д \ А А с Аб с AG UU С 1810 ps m2 A GA / € GA C GA U SUACU UC UUGAGCGGUUAGU GGGC UC UUGGUC cecucuecucu® с ИЕН PEPE PTE ЕНТО HIT sauce AG por sun UCA CUCG AG AGCCGG GCCAGGCCGCG „N АА Аб с А, 6 uu С Ina jm 6 na A GA с ба LC (GIA АСУ uc VUGAGCEGUU AGU 6666 UC UUGGUC YegoucuccccuS с АО САО ООС ОСАО СО SS INVES AG ¡ESUUCECUAS UCA CUCG AG AGCCGG GCCA SSBCCELE:, A CAG A „8 uu = 1009 (С) * 1714 un A GA / GA i 700 BAL 1665 1679 A GA A А С 6 SUACU UC UUGAGCGGUU AGU GGGC UC Vuscuc “C@GUCUGGUAC с A A AT LT TE JAUGA AG GGUUCGCUAG UCA CUCG AG AGCCGG GCCAGGCCGUG A А А с Ga 1709 AG A À „8 uc Cc 171 BIA 1689 1702 6, 1732 A GA С GA сб SUACU UC UUGAGCGGUUAGU GGNC UC UUGGUC Weacueneacaun с ИЕ РИЕНЕНТЕН en III III IN SENSE GGUUCGCUAGUCA CUCG AG AGCCGG GCCAGGCCYCG A \ A с AG CAG UU с А 1733 А BIG 1689 1703 G 173 A GA с GA CGA ï 6 РАСО UC UUGAGCGGUUAGU 6666 UC VUGGUCUCGGUCUGGCGU р О ТА AI JAUGA AG GGUUCGCUAGUCA CUCG AG AGCCGG GCCAGGCCGCG A NA A с AG CAG uu с А 1800 A FOS 1692 1708 1736 сс U G SUACU uc UUGAGCGGUU AGU GGGC UC UUGGUC CGGU CUGGCGU р IM ЗА TITRE JAUGA AG eee UCA CUCG AG GGCCGG GCCA 6660656, C \ А се Аб СА UU 1803 HEL Maen 1691 6 с GA С SUACU “uc LUGAGCGGUU AGU GGGC PEEP UP PEPE I НЕ SAS AS GGUUCGCUAG UCA CUCG 1720 GA U 4 G UC UUGGUC CGGUCUGGUAC ¢ A LACT AG AGCCGG GCCAGECCEUG A AG \ A Со ВАС А UC с 1788 ОМА 1654 1678 GG im A PER Rea “MG NN: ge 66 A А FUACU UC UUGAGCGG UC GAG UCGGAVU os vee a A PETE CUTE НЕ EEE | 6 YAUGA AG GGUUCGCUAG CUC AGCUUGA AGAGC АСС GCG SADA A KA UA 173 G GG АА ONC Ve 1700 G 170 A GA / Сс GA A СА / u SUACU UC UUGAGCGGUUAGU GGGC UC UUGGUC cecucuccucu. с ОВ A Ne TRUCK AG GGUUCGCUAG УСА CUCG AG AGCCGG BECA SGCCAC © с AG CAG 1797 A ox 1658 1672 1702 A GA с GA сс у 6 SUACU uc VUGAGCGGUU AGU 6666 UC вос CGGUCUGGCAC С IATA TAUGA AG RG CUUCECURGIUCR CUCG AG AGCCGG GCCAGGCCGUG A A с AG A A, 6 vu с 1768 RAD 1718 6, „104 A GA / CGA се 5 UACU UC UUGAGCGGUUAGU GGGC UC Uuscuc cecucueccau® с NI JAUGA AG GGUUCGCUAG UCA CUCG AG AGCCGG GCCAGGCCECG LÀ ue \ A А C AG An 6 UU 1815 SIP 1684 1698 A С GA ! 7 сб 6 SUACU uc VUGAGCGGUU AGU GGGC UC vec veecucuceccu с PEEP UE EEE TEE Pee НН an AG AU QUEUE UCA CUCG AG AGCCGG GCCA GGCCGCG, A C AG СА, 6 uc c 1795 С GA ЧАСУ uc VUGAGCGGUUAGU GGGC UC vuGcuc” вби cusscau® (3 INM TO А Th Aves AG Pr Е UCA CUCG AG AGCCGG GCCA LULU A с AG) Oy (CHANG UU len A FIG. 4. A-C. Secondary structure representations for stem 49 in the 18S rRNA gene for 21 taxa. 232 100 73 87 62 100} 92 5 100 65 100 92 DUTRA-CLARKE ЕТ AL. Monodonta labio Thais clavigera Aplysia sp. Siphonaria algesirae Oxyloma sp. Athoracophorus bitentaculatus Omalonyx matheroni Limicolaria kambeul Helix aspersa Balea biplicata Bakerilymnaea cubensis Lymnaea glabra Lymnaea stagnalis Stagnicola palustris Fossaria truncatula Lymnaea auricularia Radix peregra Biomphalaria alexandrina Biomphalana glabrata Laevilocaulis alte Onchidella celtica ELASMOGNATHA STYLOMMATOPHORA EUTHYNEURA Lymnaeidae Planorbidae SYSTELOMMATOPHORA FIG. 5. 50% majority-rule bootstrap consensus tree from parsimony analysis based on 18S rRNA gene se- quences. The numbers above nodes indicate bootstrap values from 500 replicates. from their analysis. Moreover, Bargues 8 Mas-Coma (1997) discriminated species of the family Lymnaeidae based on the phyloge- netic information of the highly variable stem E10-1. Our alignment did not corroborate the find- ings of Kenchington et al. (1994). They de- scribed an eight base pair insertion located in a highly variable domain of the 18S rRNA gene (V-4, helix E23-5) as unique for the pul- monate family Achatinidae. For example, we found the same eight base pairs to be shared by the pulmonate families Succineidae, Siphonariidae, and Lymnaeidae; and by Atho- racophoridae, Helicidae and Onchididae (with only one change). Their findings could be considered a problem of taxon sampling, or could be simply an alignment problem. We re- ported only four of the nine highly variable subdomains of the 18S rDNA as referred by Kenchington et al. (1994) as phylogenetically important, because only those four were found to be highly variable and therefore more informative among the studied Euthyneura. Rosenberg et al. (1994) stressed the impor- tance of the alignment and of other factors such as bias in nucleotide composition, and the ratio of transition to transversion. Vawter & Brown (1993) pointed out there are no con- sistent transition to transversion biases in the 18S rRNA gene. They found a C to T or vice- versa transition to be common, and also state there are phylogenetic biases in base compo- sition: Vertebrates are more G/C rich overall (1.24) than are the invertebrates (0.96). We also found changes in T to C (or C to T) to be common for our data set. The MP and ML analysis support unam- PHYLOGENETIC STUDY OF THE FAMILY SUCCINEIDAE 100 81 69 99 22 100 7 43 64 68 100 61 34 91 100 72 26 93 233 Monodonta labio Thais clavigera Aplysia sp. Biomphalaria glabrata Planorbidae Biomphalaria alexandrina Bakerilymnaea cubensis Lymnaea glabra Lymnaea stagnalis Stagnicola palustris Lymnaeidae Fossaria truncatula Lymnaea auricularia Radix peregra ete EUTHYNEURA Limicolaria kambeul Helix aspersa Balea biplicata STYLOMMATOPHORA Oxyloma sp. Athoracophorus bitentaculatus ELASMOGNATHA Omalonyx matheroni Siphonaria algesirae Laevilocaulis alte SYSTELOMMATOPHORA Onchidella celtica FIG. 6. Maximum likelihood tree based on 18S rRNA gene sequences. Numbers on the nodes represent bootstrap values out of 500 replicates. biguously the clade Elasmognatha on the main lineage of the pulmonates. However, both trees fail to support the monophyly of Succineidae. Perhaps there are undetectable homoplasies or hidden mutations for the Atho- racophoridae that make their relationship with one of the Succineidae taxon artificially strong. Winnepemninckx et al. (1995) consid- ered an Euthyneura node without a represen- tative of the Opistobranchia. Thollesson (1999), based on the 16S rRNA gene data, found Opistobranchia was not monophyletic and suggested abandoning the taxon Opisto- branchia. This suggestion corroborates the hypothesis of Tillier et al. (1992, 1994) that Aplysia and Pulmonata are more closely re- lated to each other than is Aplysia to other nudibranchs. Tillier et al. (1992). using the D1 domain of the 28S rRNA. also showed that the clade formed by Succinea (Succineidae) and Aneita (Athoracophoridae) was in a sister group position to all other pulmonates, ba- sommatophorans, and stylommatophorans, or it could be placed inside the stylom- matophorans. Our trees show the clade formed by Succineidae and Athoracophoridae taxa was positioned among the derived sty- lommatophorans and that they were not more related to Achatinidae or Helicidae than are those late taxa related to the systelom- matophorans, represented here by L. alte and O. celtica. Emberton et al. (1990), using par- tial sequence from 28S rRNA, discussed the position of basommatophorans in relation to 234 DUTRA-CLARKE ET AL. Monodonta labio Thais clavigera Aplysia sp. Siphonaria algesirae Balea biplicata 60 Oxyloma sp. 99 76 93L_ Omalonyx matheroni 61 fr Helix aspersa STYLOMMATOPHORA Athoracophorus bitentaculatus ELASMOGNATHA Limicolaria kambeul EUTHYNEURA Bakerilymnaea cubensis Fossaria truncatula 100 100 Lymnaea auricularia Radix peregra Lymnaeidae Stagnicola palustris spp” Lymnaea glabra Lymnaea stagnalis 100 Biomphalaria alexandrina Biomphalaria glabrata 88 Laevilocaulis alte Onchidella celtica | Planorbidae SYSTELOMMATOPHORA FIG. 7. 50% majority-rule bootstrap consensus tree from parsimony analysis based on 18S rRNA gene se- quences excluded of bulges and unpaired nucleotides of the highly variable regions. The numbers above nodes indicate bootstrap values from 500 replicates. the stylommatophorans. Basommatophora is paraphyletic, with lymnaeids + planorbids being sister to stylommatophorans when we conclude Siphonariidae does not represent the basommatophorans. Hubendick (1978) pointed out for the possibility of the basomm- matophorans to do not constitute a single phylogenetic unit. As Solem & Yochelson (1979) point out, there are problems in estab- lishing a direct relationship between basom- matophorans and stylommatophorans. They say the accepted position of the basom- matophoran as a stem group to various sty- lommatophoran is not supported by the his- torical sequence of the fossil record. Our results demonstrate the power of resolution of the complete sequences of 18S rDNA for studies of molluscan family and order rela- tionships, which includes from conserved to highly variable domains. PHYLOGENETIC STUDY OF THE FAMILY SUCCINEIDAE 235 ACKNOWLEDGEMENTS We thank Kenneth Emberton for useful comments on an early version of this manu- script, Thomas Wilke for very helpful com- ments on a later versions and Christine Spol- sky from the Department of Malacology of the Academy of Natural Sciences for helping with analysis of the data. We also thank Daniel Bramblett of the Department of Biochemistry and Molecular Genetics of the University of Alabama at Birmingham for his assistance with the secondary structure. K. M. Kjer of Rutgers University kindly assisted with the method of aligning sequence data including secondary structure notation. A. V. C. D.-C. is indebted to the Brazilian Council of Scientific Research (CNPq) for a doctorate scholarship. This work was supported by funds of the Betz Chair to J. R. S. and to George M. Davis, N. 1. H. grant Al-11373. LITERATURE CITED ADAMKEWICZ, S. L., M. G. HARASEWYNCH, J. BLAKE, D. SAUDEK & C. J. 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Revised ms. accepted 26 November 2000 MALACOLOGIA, 2001, 43(1-2): 237-311 DEEP-SEA CRYPTOBRANCH DORID NUDIBRANCHS (MOLLUSCA, OPISTHOBRANCHIA) FROM THE TROPICAL WEST PACIFIC, WITH DESCRIPTIONS OF TWO NEW GENERA AND EIGHTEEN NEW SPECIES Ángel Valdés Department of Invertebrate Zoology and Geology, California Academy of Sciences, Golden Gate Park, San Francisco, СА 94118, U.S.A.; avaldes Ocalacademy.org ABSTRACT The study of a large collection of cryptobranch dorid nudibranchs from deep waters in New Caledonia and the Philippines revealed the presence of Austrodoris kerguelenensis (Bergh, 1884); 18 new species belonging to the genera Cadlina, Austrodoris, Geitodoris, Discodoris, Peltodoris, Paradoris, Diaulula, Rostanga, Sclerodoris, Baptodoris and Dendrodoris; and two previously undescribed genera, Goslineria and Pharodoris. The anatomy of all these species, in- cluding the digestive, reproductive, and nervous system, are studied in detail. All these species are clearly distinguishable from other members of their genera. Most of the species have a pale, simple background coloration, and two of them lack eyes. Both character- istics seem to be adaptations to living in deep waters. Other deep-water Atlantic and Pacific species of dorid nudibranchs have similar adaptations. The two new genera are characterized by the presence of large copulatory spines, numerous flexible spines in Goslineria, and two solid, bifid spines in Pharodoris. No other cryptobranch dorid genera previously described have simi- lar copulatory spines. Some of the species here described belong to genera previously reported from cold or tem- perate waters, such as Austrodoris, Cadlina and Diaulula. Most of the species belong to genera that are widespread in either cold, temperate or tropical waters (Rostanga, Paradoris, Geitodoris and Baptodoris), and only two belong to exclusively tropical genera (Sclerodoris and Den- drodoris). Vicariant events and vertical dispersal could explain the processes of speciation and the origin of these deep-water species. Key words: Mollusca, Nudibranchia, Doridina, deep-water, Pacific Ocean. INTRODUCTION In contrast to other groups of mollusks with a well-documented deep-sea occurrence and numerous described deep-water species, very little is known about deep-sea opistho- branch mollusks, and particularly about deep- sea dorid nudibranchs. To date, only three papers dealing with deep Atlantic species of dorids have been published. The first is Bouchet (1977), who described five new species and a new genus of cryptobranch dorids, including phyllidiids, from the northeast Atlantic. Later, Valdés 8 Ortea (1996) described two new deep-sea phyllidiids, also from the northeast Atlantic, and Valdés 8 Bouchet (1998a) described an abyssal new genus of Corambidae from the Norwegian Sea. The first report on deep-water Indo-Pacific dorids was Bergh's (1884) description of Ba- thydoris abyssorum Bergh, 1884, collected at 4,435 m depth off New Ireland, Papua New 237 Guinea, during the Challenger Expedition. Bergh (1884) also described Archidoris aus- tralis Bergh, 1884, collected off the Kerguelen Island at 173 m depth. Other dorids studied in that paper were collected from shallow water. Subsequently, García et al. (1993) reported Austrodoris kerguelenensis (Bergh, 1884) from Antarctic deep waters. The genus Bathy- doris is collected from deep or cold shallow waters, and it is the only genus of dorid nudi- branchs that is consistently known from the deep sea (Wágele, 1989; Baranets, 1993; Valdés & Bertsch, 2000). There had not been a significant collecting effort of deep-sea opisthobranchs in the Indo- Pacific until the French Musorstom Expedi- tions. Three papers on dorid nudibranchs have been produced from the material col- lected in these expeditions: Valdés (2001) de- scribed 12 new species of Phyllidiidae (gen- era Phyllidia and Phyllidiopsis) from deep waters around New Caledonia; Valdés 8 Gosliner (in press) described four new 238 VALDÉS species and a new genus of caryophyllidia- bearing dorids from New Caledonia and the Philippines; and Fahey 4 Gosliner (2000) de- scribed three new species of the genus Hal- gerda from these two areas and from Fiji. Despite this recent proliferation of papers, there are still numerous genera and families of dorids never reported from the deep sea. The present paper deals with the bulk of the collection of cryptobranch dorids collected during the Musorstom Expeditions; it consti- tutes the first attempt to describe in detail the deep-water biodiversity of Indo-Pacific dorid nudibranchs. MATERIALS AND METHODS The material examined was collected during several French scientific expeditions to the southwest Pacific Ocean (Fig. 1) and the Philippines between the years 1981 and 1994, organized by the Institut de Recherche pour le Développement (IRD, formerly ORSTOM) and the Muséum National d'Histoire Naturelle, Paris. All specimens, including the type mate- rial, are deposited at the Laboratoire de Biolo- gie des Invertebres Marins et Malacologie of the Museum National d'Histoire Naturelle, Paris (MNHN), the Department of Invertebrate Zoology and Geology of the California Acad- emy of Sciences (CASIZ) and the Australian Museum, Sydney (AM). Registration numbers are not used in MNHN, and individual lots and specimens are identified by the unique combi- nation of the station number and cruise acronym. Descriptions of living animals are based on photographs or notes by collectors. All speci- mens available were dissected, and morpho- logical examination was facilitated by making a dorsal incision. The internal features were examined and drawn using a dissecting mi- croscope with a camera lucida. A portion of the mantle was critical-point dried for the Scanning Electron Microscope (SEM). Spe- cial attention was paid to the morphology of the reproductive system, digestive system, and central nervous system. The numbering of the nerves in the central nervous system was determined according to their relative po- sition on the ganglia. The buccal mass was re- moved and dissolved in 10% sodium hydrox- ide until the radula was isolated from the surrounding tissue. The radula was then rinsed in water, dried, and mounted for exam- ination with the SEM. DESCRIPTIONS Genus Cadlina Bergh, 1879 Type Species: Doris repanda Alder & Han- cock, 1842, by monotypy [= Cadlina lae- vis (Linnaeus, 1767)]. Cadlina abyssicola Valdés, new species (Figs. 2A, 3, 4) Material Examined HOLOTYPE: East of New Caledonia, Bathus 1 Expedition, stn. CP670 (20%54'S, 165°53'E), 394-397 m, 14 March 1993, 17 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). PARATYPES: New Caledonia, Musorstom 4 Expedition, stn. DW222 (22°38'S, 167°37'E), 535-560 m, 29 September 1985, one specimen 13 mm preserved length, dis- sected, leg. P. Bouchet and B. Richer de Forges (MNHN); stn. CC247 (22%09'S, 167°13’E), 435-460 m, 4 October 1985, one specimen 20 mm preserved length, dis- sected, leg. P. Bouchet and B. Richer de Forges (MNHN). lle des Pins, south of New Caledonia, SMIB 8 Expedition, stn. DW195 (22°59'S, 168°21'E), 508-514 m, 1 February 1993, one specimen 20 mm preserved length, dissected, leg. P. Bouchet and B. Richer de Forges (MNHN). South of New Caledonia, Bathus 2 Expedition, stn. CP735 (23°02'S, 166°56'E), 530-570 m, 13 May 1993, one specimen 15 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). Norfolk Ridge, New Caledonia, Bathus 3 Ex- pedition, stn. CP831 (23%04'S, 166°56’E), 650-658 m, 30 November 1993, one speci- men 13 mm _ preserved length, leg. P. Bouchet, B. Richer de Forges and A. Warén (MNHN); stn. CP846 (23%03'S, 166°58’E), 500-514 m, 1 December 1993, one specimen 14 mm preserved length, leg. P. Bouchet, B. Richer de Forges and A. Warén (MNHN); stn. CP847 (23°03'S, 166°58’E), 405-411 m, 1 December 1993, one specimen 15 mm pre- served length, leg. P. Bouchet, B. Richer de Forges and A. Warén (MNHN). Southwest of New Caledonia, Halipro 1 Expedition, stn. CP868 (21°14'S, 165°55’E), 430-455 m, 23 March 1994, one specimen 12 mm preserved length, leg. B. Richer de Forges (MNHN); stn. CP869 (21°14'S, 165°55’E), 450-490 m, 23 March 1994, three specimens 11-16 mm pre- served length, leg. B. Richer de Forges (CASIZ 121095). North of New Caledonia, DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 239 Espíritu 16° miele ne ale sele ne TS CES ee ьеынныи > Récifs : d'Entrecasteaux Santo © ‘0 a ER ER WEN $, . A Port-Vila 18°1---------- ----- ; AT» { pro a Récifs de l'Astrolabe 224 DAS: NEE : 162° 164° 166° , \ AN 20% С REN EN ent LES я , чение ничьи = еее еее CE CE , 0 \ 2 x > Û AN мно eu nn an nenn gr u nenn un nn ne 11107 168° FIG. 1. Map of the New Caledonia region, where most of the samples were collected. Bathus 4 Expedition, stn. DW941 (19°02'S, 163°27'E), 270 m, 8 August 1994, one speci- men 14 mm preserved length, leg. B. Métivier and В. Richer de Forges (ММНМ). External Morphology The body is somewhat elevated, oval (Fig. 2A). The dorsum is covered with simple, con- ical tubercles. Some of them, more sparsely arranged, are much larger than the others. There are several, simple and isolated mantle glands around the mantle margin. The perfoli- ate rhinophores are composed of 30 lamellae. There are seven bipinnate branchial leaves. Ventrally, the oral tentacles are short and grooved (Fig. 3F). The anterior border of the foot is grooved but not notched. The mantle margin is about as wide as the foot. Based on the field notes, the background color of the living animals is uniformly white to cream. The apices of the larger tubercles are yellow. The rhinophores and gill have the same color as the dorsum. The mantle glands are yellow in the preserved specimens. Anatomy The posterior end of the oral tube has two strong retractor muscles (Fig. 3E), which are 240 VALDES FIG. 2. Preserved specimens. A. Cadlina abyssicola, new species, paratype from Musorstom 4 (stn. CC247); scale bar = 5 mm. B. Austrodoris kerguelenensis, Biocal (stn. DW33); scale bar = 2 mm. C. Austrodoris caeca, new species, paratype from Halipro 1 (stn. CH876); scale bar = 5 mm. D. Pharodoris diaphora, new species, holotype; scale bar = 2 mm. E. Pharodoris philippinensis, new species, holotype; scale bar = 2 mm. F. Austrodoris laboutei, new species, holotype; scale bar = 2 mm. G. Geitodoris pallida, new species, holo- type; scale bar = 1 mm. H. Discodoris achroma, new species, paratype from Bathus 3 (stn. CP832); scale bar = 2 mm. |. Discodoris sp.; scale bar = 10 mm. FIG. 3. Cadlina abyssicola, new species, paratype from SMIB 8 (stn. DW195). A. Dorsal view of the anatomy, scale bar = 1 mm. B. Reproductive system; scale bar = 1 mm. C. Detail of the ampulla and prostate con- nection: scale bar = 1 mm. D. Central nervous system; scale bar = 0.5 mm. E. Anterior portion of the diges- tive system; scale bar = 1 mm. F. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, defer- ent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pl, pleural nerve; plg, pleural ganglion; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, seminal receptacle; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. 242 VALDÉS attached to the body wall. The small, oval, muscular buccal bulb has two additional mus- cles. Two long salivary glands connect to the buccal bulb at the sides of the esophageal junction. The buccal bulb is one-half as long as the oral tube. The jaws consist of numer- ous thin, unicuspid elements about 20 um long (Fig. 4D). The radular formula is 59 x (45.1.45) in a 20 mm preserved length speci- men. The rachidian teeth have 4-5 elongate denticles, none of them clearly larger than the others (Fig. 4A). The inner lateral teeth are hamate, having a single cusp and a series of denticles. The innermost lateral tooth has 3-4 short denticles on the inner side and 6-7 short, blunt denticles on the outer side. The following teeth have 7-10 blunt denticles only on the outer side. The teeth increase in size gradually towards the medial portion of the half-row. The mid-lateral teeth are hamate, with a long cusp, and they lack denticles (Fig. 4B). The outermost teeth have 10-12 small denticles (Fig. 4C). The esophagus is long and connects directly to the digestive gland. The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 3C). The oviduct enters the female glands near the center of the mass. The prostate is tubular and elongate (Fig. 3B); it narrows and ex- pands again into the short deferent duct. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. The vagina is long and wide. At its proximal end, the vagina connects to the large, irregular bursa copulatrix. From the distal end of the vagina, near the opening, a short duct con- nects to the seminal receptacle and the uter- ine duct. The bursa copulatrix is about ten times larger than the seminal receptacle. In the central nervous system (Fig. 3D), the cerebral and pleural ganglia appear to be par- tially fused and distinct from the pedal gan- glia. There are four cerebral nerves leading from each cerebral ganglion, and two pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with two nerves leading from each. The pedal and parapedal commissures are enveloped to- gether with the visceral loop along all their length. The circulatory system (Fig. 3A) consists of a heart and a single blood gland situated over the central nervous system. Etymology From the Ancient Greek abyssos (deep) and the Latin cola (dweller), in reference to the habitat of this species, used as a noun in apposition. Remarks Cadlina abyssicola is clearly distinguish- able from other species of the genus. All the species previously described have denticu- lated mid-lateral teeth (Rudman, 1984), whereas C. abyssicola has smooth mid-lat- eral teeth. Also, species of Cadlina, except Cadlina dubia Edmunds, 1981, have bifid jaw rodlets (Rudman, 1984), whereas in C. abyssicola all of them are simple. Cadlina dubia, described from Ghana, differs from C. abyssicola in having much shorter radular teeth, all bearing strong denticles (Edmunds, 1981). Cadlina willani Miller, 1980, from New Zealand, is the species with the closest geo- graphic range to C. abyssicola. It is a white species with a mid-dorsal yellow stripe and a yellow band around the mantle margin. It is very different from C. abyssicola, which is white with yellow spots. Moreover, the mor- phology of the radular teeth is very different between these two species. The inner lateral teeth of C. abyssicola are narrow and elon- gate and have 6-7 short denticles on the outer side of the cusp, whereas those of C. willani are shorter and have only 3 denticles on the outer side (Miller, 1980). Also, the mid- lateral teeth of C. abyssicola are smooth, whereas in C. willani the mid-lateral teeth have 7 strong denticles. Cadlina nigrobranchiata Rudman, 1985, from southern Australia is clearly distinguish- able from C. abyssicola in having a black gill and black rhinophores (Rudman, 1985). Also, the radular morphology of these two species is very different. The rachidian teeth of C. ni- grobranchiata are much shorter than those of C. abyssicola. Moreover, the mid-lateral teeth of C. abyssicola are smooth, whereas in C. nigrobranchiata they have several strong den- ticles. Schródl (2000) reviewed the species of Cadlina from South America and Antarctica. All of them are clearly distinguishable from C. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 243 FIG. 4. Cadlina abyssicola, new species, paratype from SMIB 8 (stn. DW195). A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 43 um. D. Jaw ele- ments; scale bar = 30um. abyssicola in their radular morphology and anatomy. These five species — Cadlina ker- guelensis Thiele, 1912; C. sparsa (Odhner, 1921); C. magellanica Odhner, 1926; C. affi- nis Odhner, 1934; C. georgiensis Schrödl, 2000 — have denticulate mid-lateral teeth, whereas they are smooth in C. abyssicola. In C. affinis and C. magellanica, the seminal re- ceptacle inserts in the middle of the vagina, in C. kerguelensis and C. sparsa it inserts near the bursa copulatrix, whereas in C. georgien- sis and C. abyssicola the insertion is closer to the opening of the vagina. However, in C. abyssicola the seminal receptacle is more dis- 244 VALDÉS tal than in C. georgiensis and it is clearly stalked. In addition, the prostate and the am- pulla of C. abyssicola appear to be longer than those of C. georgiensis. Cadlina sp. (Fig. 5) Material Examined New Caledonia, Biocal Expedition, stn. DW14 (22°47'S, 167°14'E), 440-450 m, 30 August 1985, one specimen 5 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). External Morphology The body is somewhat elevated, oval. The dorsum is covered with simple, conical tuber- cles that are evenly arranged. The perfoliate rhinophores are composed of nine lamellae. There are five bipinnate branchial leaves. Ventrally, the oral tentacles are short and grooved. The anterior border of the foot is simple. The mantle margin is wider than the foot. The color of the living animal is unknown. The preserved specimen is uniformly white to cream. The rhinophores and gill have the same color as the dorsum. Anatomy The jaws consist of numerous thin, unicus- pid elements about 10 um long (Fig. 5D). The radular formula is 68 x (19.1.19) in a 5 mm preserved length specimen. The rachidian teeth have a large, single central cusp and 5-6 blunt denticles on each side (Fig. 5A). The inner lateral teeth are wide, hamate, hav- ing a single cusp and a series of denticles. These teeth have 2-3 denticles on the inner side and 10-12 elongate denticles on the outer side. The teeth increase in size gradu- ally towards the medial portion of the half-row. The mid-lateral teeth are hamate, with a long cusp and 7-8 large denticles only on the outer side (Fig. 5B). The outermost teeth have 5-11 denticles (Fig. 5C). The reproductive system of the single spec- imen collected was immature, so information on the genital morphology is not available. Remarks The radular morphology of this species is very distinct from other members of the genus Cadlina. The rachidian teeth have a number of small denticles on each side of the central cusp that are not present in any other species previously described (Rudman, 1984) However, since the color of the living animal is unknown, and the reproductive system of the single specimen collected is immature, | prefer not to name this species until new ma- terial becomes available. Genus Austrodoris Odhner, 1926 Type Species: Archidoris rubescens Bergh, 1898, by original designation [= Aus- trodoris kerguelenensis (Bergh, 1884)]. Austrodoris kerguelenensis (Bergh, 1884) (Figs: 2B: 6; 7) Archidoris kerguelenensis Bergh, 1884: 85- 89, pl. 1, figs. 1-12. Austrodoris georgiensis Garcia et al., 1993: 417-421, figs. 1-8, new synonymy herein. See Wagele (1990, 1993) for a complete list of synonyms. Material Examined New Caledonia, Biocal Expedition, stn. DW33 (23°10'S, 167°10'E), 675-680 m, 29 August 1985, one specimen 13 mm pre- served length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). lle des Pins, New Caledonia, SMIB 8 Expedition, stn. DW193 (25°59'S, 168°21'E), 500-508 m, 1 February 1993, two specimens 10 mm pre- served length, leg. P. Bouchet and B. Richer de Forges (MNHN). External Morphology The body is oval, almost rounded and somewhat elevated (Fig. 2B). The whole dor- sum is covered with small, elongate tubercles. In general, tubercles are larger in the center of the dorsum. Some tubercles are clearly longer and are surrounded by areas with nu- merous short tubercles (Fig. 6D). The rhinophoral and branchial sheaths are edged by several long tubercles. The large, perfoli- ate rhinophores are composed of 38 lamellae. The gill consists of six short, bipinnate branchial leaves. Ventrally, there are two short, conical oral tentacles (Fig. 7E). The anterior border of the DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 245 FIG. 5. Cadlina sp. specimen from Biocal (stn. DW14). A. Inner lateral teeth; scale bar = 15 um. B. Mid-lat- eral teeth; scale bar = 15 um. С. Outer lateral teeth; scale bar = 15 um. D. Jaw elements; scale bar = 10 um. foot is grooved but not notched. The mantle margin is wider than the foot. Based on the field notes, the color of the liv- ing animals is uniformly white, with yellowish tubercles. The rhinophores and gill are pale yellow. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 7C), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The 246 VALDES uk FIG. 6. Austrodoris kerguelenensis, specimen from SMIB 8 (stn. DW193). A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 60 um. D. Dorsal tubercles; scale bar = 430 um. buccal bulb is as long as the oral tube. The labial armature is smooth. The radular for- mula is 36 x (56.0.56) in a 10 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are hamate, hav- ing a single cusp, and they lack denticles (Fig. 6A). The teeth increase in size gradually to- wards the medial portion of the half-row (Fig. 6B). The outermost teeth are elongate and also lack denticles (Fig. 6C). The esophagus is long and connects to the stomach. The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into the short oviduct and the prostate. The oviduct connects to the female glands near the center of the mass (Fig. 7B). The prostate DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 247 FIG. 7. Austrodoris kerguelenensis, specimen from SMIB 8 (stn. DW193). A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 0.5 mm. C. Anterior portion of the digestive system; scale bar = 0.5 mm. D. Central nervous system; scale bar = 0.5 mm. E. Mouth area; scale bar = 1 mm. Ab- breviations: a, ampulla; ab, abdominal ganglion; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, re- tractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pl, pleural nerve; plg, pleural ganglion; pr, prostate; г, rhinophoral nerve; $, seminal receptacle; sg, salivary gland; st, stomach: t, oral tentacle; v, vagina. 248 VALDÉS is tubular and convoluted; it expands again into the long and wide deferent duct. The def- erent duct opens into a common atrium with the vagina. The penis is unarmed. The vagina is short and connects distally to the oval bursa copulatrix. From the bursa copulatrix leads another duct that branches into the uterine duct and the seminal receptacle. The seminal receptacle has a short, convoluted stalk. The bursa copulatrix is about three times larger than the seminal receptacle. In the central nervous system (Fig. 7D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, three pleural nerves leading from the right pleural ganglion and three from the left ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gas- troesophageal, rhinophoral, and optical gan- glia are present. The pedal ganglia are clearly separated, with three nerves leading from the left ganglion and two from the right ganglion. The pedal and parapedal commissures are enveloped together with the visceral loop along most of their length. There is an ab- dominal ganglion on the right side of the vis- ceral loop. The circulatory system (Fig. 7A) consists of a heart and two blood glands situated in front and behind the central nervous system. Remarks The external morphology and anatomy of the specimens from New Caledonia are very similar to the descriptions by Bergh (1884) and Wagele (1990) of Austrodoris kerguele- nensis, and they clearly belong to the same species. In both the Antarctic and New Cale- donia specimens, the dorsal tubercles are rounded, some of them being clearly longer, surrounded by areas with numerous short tu- bercles. Also, the reproductive system of the New Caledonia specimens has a very long, muscular deferent duct, a short tubular prostate, and a large, convoluted ampulla, identical to the description of Wágele (1990) for A. kerguelenensis. The proportions and arrangement of the bursa copulatrix and sem- inal receptacle is also very similar between the Antarctic and New Caledonia animals. Moreover, the radula of A. kerguelenensis is composed of numerous simple, hamate teeth (Wagele, 1990), very similar to those de- scribed here. According to Wagele (1990), Austrodoris kerguelenensis is the only valid species of this genus. Recently, García et al. (1993), de- scribed Austrodoris georgiensis based on a single specimen collected from South Geor- gia in the Atlantic Antarctic sector. The only difference between A. georgiensis and A. ker- guelenensis is the presence of an elongate bursa copulatrix in the former. Because other features of both nominal species (e.g., exter- nal morphology, radula, other reproductive or- gans), are identical, it is likely that the single specimen assigned to A. georgiensis is just an aberrant specimen of A. kerguelenensis. Another possibility is that the bursa copulatrix is more variable than assumed until now. Austrodoris kerguelenensis, originally de- scribed from the Kerguelen Islands, is distrib- uted throughout the Atlantic sector of the Antarctica (Wägele, 1990; Garcia et al., 1993). Bouchet (1977) recorded A. kerguele- nensis from Atlantic waters off Uruguay (as Austrodoris macmurdensis). The present record, from New Caledonia, is a consider- able range extension for this species. Austrodoris caeca Valdés, new species (Figs. 2C, 8-10) Material Examined HOLOTYPE: Banc Combe, South Pacific, Musorstom 7 Expedition, stn. CP551 (12°15'S, 177°28'W), 791-795 m, 18 May 1992, 10 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). PARATYPES: Banc Combe, South Pacific, Musorstom 7 Expedition, stn. CP552 (12°16'S, 177°28'W), 786-800 m, 18 May 1992, one specimen 12 mm preserved length, dissected, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). Banc Tuscarora, South Pacific, Musorstom 7 Expedition, stn. CP562 (11°48’S, 178°22’W), 775-777 m, 19 May 1992, one specimen 12 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN); stn. CP623 (12°34'S, 178°15’W), 1280-1300 m, 28 May 1992, one specimen 18 mm preserved length, dissected, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). South of New Caledonia, Bathus 2 Expedition, stn. CP766 (22°10'S, 166°02’E), 650-724 m, 17 May 1993, one specimen 12 mm preserved length, leg. P. Bouchet and B. Richer de Forges (CASIZ 121096). Norfolk Ridge, New Caledo- DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 249 y zu y ri fs : = zm FIG. 8. Austrodoris caeca, new species, paratype from Musorstom 7 (stn. CP552). A. Inner lateral teeth; scale bar = 60 um. B. Mid-lateral teeth; scale bar = 75 um. C. Outer lateral teeth; scale bar = 75 um. D. Dor- sal tubercles; scale bar = 200 um. nia, Bathus 3 Expedition, stn. CC848 (23°02'S, 166°53’E), 680-700 m, 1 Decem- ber 1993, two specimens 14 mm preserved length, leg. P. Bouchet, B. Richer de Forges and A. Warén (MNHN). New Caledonia, Halipro 1 Expedition, stn. CH876 (23°10’S, 166°49'E), 870-1000 m, 31 March 1994, 23 mm preserved length, dissected, leg. B. Richer de Forges (MNHN). Vanuatu, South Pacific, Musorstom 8 Expedition, stn. CP1074 (15°48'S, 167°24'E), 775-798 m, 4 October 1994, one specimen 13 mm preserved length, dissected, leg. P. Bouchet and B. Richer de Forges (MNHN). 250 VALDÉS FIG. 9. Austrodoris caeca, new species, paratype from Musorstom 8 (stn. CP1074). A. Inner lateral teeth; scale bar = 60 um. B. Mid-lateral teeth; scale bar = 100 um. C. Outer lateral teeth; scale bar = 75 um. D. Dor- sal tubercles; scale bar = 250 um. External Morphology The body is oval, almost rounded and somewhat elevated (Fig. 2C). The whole dor- sum is covered with small, rounded tubercles. Some of them are irregularly distributed and larger than the others (Figs. 8D, 9D). The rhinophoral sheaths are elevated and covered with small tubercles. The large, perfoliate rhinophores are composed of 41 lamellae. The gill consists of six short, bipinnate branchial leaves. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 251 FIG. 10. Austrodoris caeca, new species, paratype from Musorstom 7 (stn. CP552). A. Dorsal view of the anatomy, scale bar = 1 mm. B. Reproductive system; scale bar = 1 mm. C. Detail of several reproductive or- gans; scale bar = 1 mm. D. Anterior portion of the digestive system; scale bar = 1 mm. E, Central nervous system; scale bar = 0.5 mm. F, Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; b, buccal bulb; be, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart: i, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; sg, salivary gland; st, stomach; v, vagina. 252 VALDÉS Ventrally, there are no oral tentacles (Fig. 10F). The anterior border of the foot is grooved but not notched. The color of the living animals is unknown. The preserved specimens are uniformly cream or pinkish. The rhinophores and gill are pale cream. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 10D), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb is three times longer than the oral tube. The labial armature is smooth. The radular formula is 21 x (30.0.30) in a 12 mm preserved length specimen and 26 x (36.0.36) in a 13 mm preserved length speci- men. There are no rachidian teeth. The inner lateral teeth are hamate, having a single cusp, and they lack denticles (Figs. 8A, 9A). The teeth increase in size gradually towards the medial portion of the half-row (Figs. 8B, 9B). The outermost teeth are elongate but shorter than the adjacent ones, and also lack denti- cles (Figs. 8C, 9C). The esophagus is long and connects directly to the stomach. The ampulla is thin and elongate; it narrows into the gonoduct, which branches into the short oviduct and the prostate (Fig. 10C). The oviduct connects to the female glands near the center of the mass. The prostate is tubular and elongate (Fig. 10B); it narrows slightly and expands into the long, wide and strongly muscular deferent duct. The deferent duct opens into a common atrium with the vagina. The penis is unarmed and very long, about 2/3 of the ejaculatory portion of the deferent duct length. The vagina is elongate. At its proximal end, the vagina connects to the oval bursa copulatrix. From the bursa copulatrix leads another duct that connects to the semi- nal receptacle and the uterine duct. The sem- inal receptacle has a long, convoluted stalk. The bursa copulatrix is about eight times larger than the seminal receptacle. In the central nervous system (Fig. 10E), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, and two pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. There are no eyes. The pedal ganglia are clearly sepa- rated, with three nerves leading from each. The pedal and parapedal commissures are enveloped together with the visceral loop along most of their length. The circulatory system (Fig. 10A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Latin caecum (blind), in reference to the lack of eyes in this species. Remarks The genus Austrodoris is very closely re- lated to Archidoris Bergh, 1878. According to Wagele (1990), the main differences between these two genera are that in Austrodoris most of the length of the deferent duct is encased within a muscular sheath, there is no glans penis, and the seminal receptacle and bursa copulatrix insert opposite and not serially. The genus Austrodoris appears to be restricted to the Southern Hemisphere, whereas Archi- doris is mainly distributed in the Northern Hemisphere. An anatomical study and com- parison between the type species of both gen- era is necessary before their systematic sta- tus can be determined. In the meanwhile, | follow Wagele’s (1990) criteria and maintain Austrodoris as a valid genus. Austrodoris caeca clearly belongs to the genus Austrodoris in lacking a glans penis, having a very large, elongate and strongly muscular deferent duct, longer than the prostate, smooth and hamate radular teeth, and a smooth labial cuticle. Anatomically, A. caeca differs from Aus- trodoris kerguelenensis in having a seminal receptacle with a long stalk, having a much shorter ampulla, and in lacking eyes. In addi- tion, the outermost lateral teeth of A. caeca are very short compared to the mid-laterals, whereas in A. kerguelenensis they are pro- portionally larger. Austrodoris laboutei Valdés, new species (Figs. 2, 115,12) Material Examined HOLOTYPE: East of New Caledonia, Bathus 1 Expedition, stn. CP711 (21°43'S, 166°36’E), 315-327 m, 19 March 1993, one specimen 15 mm preserved length, dis- DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 253 FIG. 11. Austrodoris laboutei, new species, holotype. A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 75 um. C, Outer lateral teeth; scale bar = 43 um. D, Dorsal tubercles; scale bar = 75 um. sected, leg. P. Bouchet and B. Richer de Forges (MNHN). External Morphology The body is elongate and high (Fig. 2F). The mantle margin is very reduced. The whole dorsum is covered with small, rounded tubercles, all of them similar in size (Fig. 11D). The rhinophoral sheaths are low, situated very anteriorly, and covered with small tuber- cles. The large, perfoliate rhinophores are composed of 19 lamellae. The gill sheath has a wide opening and it is situated very posteri- orly. The gill consists of seven long, tripinnate branchial leaves. Ventrally, the oral tentacles are wide, short, and grooved (Fig. 12E). The anterior border of 254 VALDÉS FIG. 12. Austrodoris laboutei new species, holotype. A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 0.5 mm. C. Anterior portion of the digestive system; scale bar = 1 mm. D. Central nervous system; scale bar = 0.5 mm. E. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; ag, accessory gland; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and di- gestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; sg, salivary gland; st, stom- ach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 255 the foot is grooved but not notched. The foot is wide. Based on the field notes, the color of the liv- ing animals is cream grayish. The dorsum is covered with a number of small, brown spots. The rhinophores are pale brown, and the gill is cream grayish with brown spots. The pre- served specimen is uniformly cream. Anatomy The posterior end of the oral tube has four strong retractor muscles (Fig. 12C), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. Two long salivary glands are attached to the buc- cal bulb at the point where the esophagus connects. There is another glandular struc- ture that connects ventrally to the buccal bulb at the point where the oral tube opens into the buccal bulb. The buccal bulb is one-third the length of the oral tube. The labial armature is smooth. The radular formula is 15 x (16.0.16) in a 16 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are hamate, having a very long, thin cusp that lacks denticles (Fig. 11A). The teeth increase in size suddenly towards the medial portion of the half-row. The mid-lateral teeth are long, having a short, conical cusp, and they lack denticles (Fig. 11B). The outermost teeth are small and also lack denticles (Fig. 11C). The esophagus is long and connects directly to the stomach. The ampulla is wide and convoluted; it nar- rows into the gonoduct, which branches into the long oviduct and the prostate (Fig. 12B). The oviduct connects to the female glands near the center of the mass. The prostate is tubular and elongate; it narrows slightly and expands again into the wide deferent duct. The deferent duct opens into a common atrium with the vagina. The penis occupies the whole ejaculatory portion of the deferent duct. The vagina is short. At its proximal end, the vagina connects to the irregular bursa copulatrix and to another short, wide duct that connects to the seminal receptacle and the uterine duct. The seminal receptacle has a short stalk. The bursa copulatrix is about eight times larger than the seminal receptacle. In the central nervous system (Fig. 12D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglion, and three pleural nerves lead from each pleural ganglion. The buccal gan- glia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with three nerves leading from each. The pedal and parapedal commissures are enveloped to- gether with the visceral loop. The circulatory system (Fig. 12A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology After Pierre Laboute, who photographed specimens studied in this paper. Remarks Austrodoris laboutei belongs to the genus Austrodoris in lacking a glans penis, having a strongly muscular deferent duct, smooth and hamate radular teeth, and a smooth labial cu- ticle. Other features of this species that are also present in members of the genus Aus- trodoris are short, non-digitiform oral tenta- cles and simple dorsal tubercles. However, the deferent duct of A. /aboute/ is much shorter than in other species of the genus, and there is a gland that connects the oral tube and the buccal bulb and an accessory gland in the reproductive system, both of which are absent in other members of the genus. The allocation of A. /aboutei in the genus Austrodoris is thus provisional, until a phylogenetic analysis of other related species will allow a more precise definition of the di- agnostic characters of the genera Archidoris and Austrodoris. Austrodoris laboutei is clearly distinguish- able from other species of the genus by the body shape, with a very reduced mantle mar- gin. The coloration of this species, cream grayish with small brown spots is different from that of A. kerguelenensis, which is uni- formly white with yellowish tubercles. Anatomically, A. Jaboutei differs from Aus- trodoris kerguelenensis and A. caeca in hav- ing an accessory gland in the reproductive system, a much longer prostate, and a very large and irregular bursa copulatrix. In addi- tion, the innermost and outermost radular teeth of A. laboutei are very long and thin compared to those of other species of the genus. On the contrary, the mid-laterals of A. laboutei are stronger than those of A. kergue- lenensis and A. caeca. 256 VALDÉS Genus Goslineria Valdés, new genus Type Species: Goslineria callosa Valdés, new species, here designated. Diagnosis Body elevated, oval, stiffened by numerous integumentary spicules. Dorsum covered with simple, conical tubercles. Anterior border of the foot grooved, not notched. Labial cuticle unarmed. Radula without rachidian teeth. Inner and mid-lateral teeth simple, hamate. Outermost lateral teeth with numerous, small denticles. Prostate flattened, flattened. Penis unarmed. Genital atrium with several large sacs, each containing a long, simple, flexible spine. Etymology Dedicated to Terry Gosliner, who made in- valuable comments on the manuscript and provided financial support for this study. Remarks The presence of several sacs in the repro- ductive system, each containing a flexible cop- ulatory spine, is the most distinctive feature of this new genus. Species of Paradoris may also have sacs with copulatory spines associated with the atrium, but in this genus the spines are rigid. Besides, species of Paradoris have jaws, armed with numerous denticles, whereas in Goslineria the labial cuticle is smooth. The new genus Pharodoris has two large, rigid copulatory spines. However, Pharodoris is different from Goslineria in having bifid spines, a highly elevated branchial sheath, and a tu- bular prostate. Other genera with copulatory spines are Hoplodoris, Asteronotus, Jorunna, and Sclerodoris, but in all those the spines are rigid and situated in the accessory gland, not in separate sacs (Gosliner & Behrens, 1998; Valdés & Gosliner, in press). Moreover, Jorunna and Sclerodoris have caryophyllidia, which are absent in Goslineria, Asteronotus, Pharodoris and Paradoris. Goslineria callosa Valdés, new species (Figs 13A, 14, 15, 16B) Material Examined HOLOTYPE: Philippines, Musorstom 2 Ex- pedition, stn. CP41 (13°16’N, 122°46’E), 166-172 m, 25 November 1980, 21 mm pre- served length, leg. P. Bouchet (MNHN). External Morphology The body is elevated and oval (Fig. 13A). The dorsum is covered with simple, conical tu- bercles (Fig. 14D). Some tubercles are clearly larger than the others. The perfoliate rhino- phores are each composed of 22 lamellae. There are seven tripinnate branchial leaves. The rhinophoral and branchial sheaths are somewhat elevated, bearing numerous tu- bercles. Ventrally, there are no oral tentacles (Fig. 15F). The anterior border of the foot is grooved, not notched. The mantle margin is as wide as the foot. Based on the photographs, the color of the living animal is uniformly pale cream. The rhinophores and gill have the same color as the dorsum. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 15E), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb tube is about twice as long as the oral tube. The labial cuticle is smooth. The radular formula is 39 x (42.0.42) in a 21 mm preserved length specimen. There are no rachidian teeth. The inner and mid-lateral teeth are hamate, having a single cusp, and they lack denticles (Fig. 14A). The teeth in- crease in size gradually towards the medial portion of the half-row (Fig. 14B). The outer- most lateral teeth are also hamate and have numerous, small denticles (Fig. 14C). The esophagus is very long, convoluted, and con- nects directly to the stomach (Fig. 15A). The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 15C). The oviduct enters the female glands near the nidamental opening. The prostate is large and flattened (Fig 15B); it narrows and expands again into the long, wide and muscular defer- ent duct. The deferent duct opens into a com- mon atrium with the vagina. The atrium has six long sacs, each one containing a long, flexible copulatory spine (Figs. 15B, 16B). The largest copulatory spine reaches up to 1.5 mm in length. The penis is unarmed. The vagina is very wide. At its proximal end, the DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 257 FIG. 13. Living animals. A. Goslineria callosa, new genus and species, holotype. B. Paradoris araneosa, new species, paratype from SMIB 8 (stn. DW182-184), photo J. L. Menou. C. Paradoris imperfecta, new species, holotype, photo P. Laboute. D. Dendrodoris brodieae, new species, paratype from Спаса! 2 (stn. DW81), photo P. Laboute. vagina connects to the large and oval bursa copulatrix. From the vagina leads another very short duct that connects to the seminal receptacle and the long uterine duct. The bursa copulatrix is about five times larger than the seminal receptacle. In the central nervous system (Fig. 15D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglia, and two pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with four (right) or three (left) nerves leading from them. The pedal and parapedal commissures are enveloped to- gether with the visceral loop along all their length. The circulatory system (Fig. 15A) consists of a heart and a blood gland situated over the central nervous system. Etymology From the Latin callosus (hard skin), in ref- erence to the hard texture of the mantle of this species. Remarks This species is clearly distinguishable from other dorids previously described by the pres- ence of several, flexible copulatory spines in the reproductive system. Genus Pharodoris Valdés, new genus Type Species: Pharodoris diaphora Valdés, new species, here designated. 258 VALDÉS FIG. 14. Goslineria callosa, new genus and species, holotype. A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 75 um. C. Outer lateral teeth; scale bar = 43 um. D. Dorsal tubercles; scale bar = 250 um. Diagnosis Body elevated, oval, stiffened by numerous integumentary spicules. Dorsum covered with simple, conical tubercles. Branchial sheath very elevated. Anterior border of the foot grooved, not notched. Oral tentacles short. Labial cuticle unarmed. Radula without rachidian teeth. Inner and mid-lateral teeth simple, hamate. Outermost lateral teeth with numerous, small denticles. Prostate long, tubular. Penis unarmed. Genital atrium with DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 259 FIG. 15. Goslineria callosa, new genus and species, holotype. A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 1 mm. C. Detail of several reproductive organs; scale bar = 1 mm. D. Central nervous system; scale bar = 0.5 mm. E. Anterior portion of the digestive system; scale bar = 1 mm. F, Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; d, deferent duct; e, esophageal ganglion; f, female glands; 9, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, re- tractor muscle; 0, esophagus; ot, oral tube; p, pedal nerve; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, seminal receptacle; sa, copulatory spine sac; sg, salivary gland; sp, copulatory spine; st, stomach; v, vagina. 260 VALDÉS Ae 4 DET ER EN 4 us FIG. 16. Copulatory organs. A. Solid structures in the accessory gland of Geitodoris pallida, new species, holotype. B. Copulatory spine of Goslineria callosa, new genus and species, holotype. two large glands, each containing a long, bifid, rigid spine. Etymology From the Ancient Greek pharos (light- house), in reference to the shape of the branchial sheath, that resembles a lighthouse tower. Remarks The new genus Pharodoris is clearly distin- guishable from other genera of cryptobranch dorids previously described. lt has several unique features, such as a very elevated branchial sheath and the presence of two large glands in the atrium, armed with bifid, rigid copulatory spines. The presence of simple dorsal tubercles, a radula lacking rachidian teeth and having sim- ple, hamate lateral teeth (except the outer- most), absence of labial armature, presence of a tubular prostate, and the anterior border of the foot grooved but not notched, relate Pharodoris with Archidoris and Austrodoris. However, these two genera lack accessory glands with copulatory spines, and the branchial sheath is not so elevated (Wagele, 1990; Valdés & Gosliner, in press). Pharodoris diaphora Valdés, new species (Figs 2D, 17, 18, 19C) Material Examined HOLOTYPE: East of New Caledonia, Bathus 1 Expedition, stn. CP707 (21°43'S, 166°36’E), 347-375 m, 19 March 1993, 19 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). PARATYPES: New Caledonia, Musorstom 4 Expedition, stn. DW231 (22°34'S, 167°10’E), 75 m, 1 October 1985, one speci- men 7 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). East of New Caledonia, Bathus 1 Expedition, stn. CP710 (21°43'S, 166°36'E), 320-386 m, 19 March 1993, one specimen 18 mm preserved length, leg. P. Bouchet and B. Richer de Forges (CASIZ 121097); stn. CP711 (21°43'S, 166°36’E), 315-327 m, 19 March 1993, one specimen 16 mm preserved length, dissected, leg. P. Bouchet and B. Richer de Forges (MNHN). Vanuatu, Musorstom 8 Ex- pedition, stn. DW964 (20°20'S, 169°49'E), 360-408 m, 21 September 1994, one speci- DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 261 FIG. 17. Pharodoris diaphora, new genus and species, paratype from Bathus 1 (stn. CP711). A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 75 um. C. Outer lateral teeth; scale bar = 60 um. D. Dorsal tubercles; scale bar = 300 um. men 9 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). ADDITIONAL MATERIAL: Passe Deverd, outer Barrier Reef, near Koumac, New Cale- donia (20°45.2'S, 164°15.2'E), 53 m, 22 Oc- tober 1993, 3 specimens 10-12 mm long alive, leg. W. Rudman (AM C200596). External Morphology The body is somewhat elevated, oval (Fig. 2D). The dorsum is covered with simple, con- ical tubercles. Some of them, more abundant in the center of the body are twice as large as the others. There are several spicules pro- FIG. 18. Pharodoris diaphora, new genus and species, paratype from Bathus 1 (stn. CP711). A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 0.5 mm. C. Detail of several repro- ductive organs; scale bar = 0.5 mm. D. Anterior portion of the digestive system; scale bar = 1 mm. E. Cen- tral nervous system; scale bar = 0.5 mm. F. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; b, buc- cal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; 1, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, seminal receptacle; sa, copulatory spine sac; sg, salivary gland; sp, copulatory spine; st, stomach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 263 FIG. 19. Pharodoris, new genus, copulatory spines. A. B. philippinensis, new species, holotype, apex of a spine. B. B. philippinensis new species, holotype, general view of a spine. C. B. diaphora new species, paratype from Bathus 1 (stn. CP711), general view of a spine, holotype. truding from the dorsal surface of the tuber- cles (Fig. 17D). The branchial sheath is very elevated, rising clearly over the dorsal surface of the body. The perfoliate rhinophores are composed of 30 lamellae. There are five bip- innate, very elongate branchial leaves. Ven- trally, the oral tentacles consist of two triangu- lar prolongations on each side of the buccal area (Fig. 18F). The anterior border of the foot is grooved but not notched. The mantle mar- gin is narrower than the foot. Based on the field notes, the background color of the living animals is uniformly white to pale yellow. The rhinophores and gill have the same color as the dorsum. Three additional specimens of this species, which have not been studied for this paper, have been collected from New Caledonia by Bill Rudman (pers. comm.). His manuscript notes read as follows: mantle with low rounded tubercles, spicules radiating from them; central region (visceral hump) with brown speckling between tubercles and on sides of them; tips of tubercles translucent 264 VALDÉS clear; mantle margin translucent cream with some tubercles ringed in brown but most not; general appearance of brownish visceral hump and narrow translucent cream mantle margin; most remarkable feature is the highly developed tall and rigid gill pocket, same color and tuberculation as dorsum; six simple branchial leaves, translucent clear with white edging. It is clear from this description that these specimens are conspecific with the other material here examined. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 18D), which are attached to the body wall. The large, oval, muscular buccal bulb has two additional mus- cles. Two long salivary glands connect to the buccal bulb at the sides of the esophageal junction. The buccal bulb is four times longer than the oral tube. The labial armature is un- armed. The radular formula is 34 x (39.0.39) in a 16 mm preserved length specimen. The are no rachidian teeth. The inner lateral teeth are hamate, having a single cusp and one denticle on the outer side (Fig. 17A). The teeth increase in size gradually towards the medial portion of the half-row (Figs 17B). The mid-lateral teeth are hamate, with a long cusp, and they lack denticles. The three out- ermost teeth are very small and have 10-11 small denticles (Fig. 17C). The esophagus is long and connects directly to the stomach (Fig. 18A). The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 18C). The oviduct enters the female glands near the nidamental opening. The prostate is tubular and elongate (Fig. 18B); it narrows and ex- pands again into the short deferent duct. The deferent duct opens into a common atrium with the vagina. On the atrium, near the open- ing of the vagina, there are two large glands, each one containing a bifid, rigid spine about 1 mm long (Figs. 18B, 19C). The penis is un- armed. The vagina is long and wide. At its proximal end, the vagina connects to the large and oval bursa copulatrix. From the medial part of the vagina leads the elongate seminal receptacle and the long uterine duct. In the central nervous system (Fig. 18E), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglion, and two pleural nerves lead from each pleural ganglion. The buccal gan- glia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with three nerves leading from each. The pedal and parapedal commissures are enveloped to- gether with the visceral loop along most of their length. The circulatory system (Fig. 18A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Ancient Greek diaphoros (differ- ent), in reference to the strange shape of this animal. Remarks Pharodoris diaphora is different from all species of cryptobranch dorids previously de- scribed. The characteristic, very elevated branchial sheath is a unique feature of this genus. Internally, the presence of two large glands containing bifid, rigid spines is only present in this genus. For a comparison to other members of this genus, see Remarks on Pharodoris philippinensis. Pharodoris philippinensis Valdés, new species (Figs. 2E, 19A, B, 20, 21) Material Examined HOLOTYPE: Philippines, Musorstom 3 Ex- pedition, stn. CP121 (12°08'N, 121°18’E), 73-84 m, 03 June 1985, 19 mm preserved length, dissected, leg. P. Bouchet and M. P. Triclot (MNHN). External Morphology The body is somewhat elevated, oval (Fig. 2E). The dorsum is covered with simple, con- ical tubercles. Some of them, more abundant in the center of the body, are twice as large as the others. The branchial sheath is very ele- vated, rising clearly over the dorsal surface of the body. The perfoliate rhinophores are each composed of 36 lamellae. There are five bip- DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 265 FIG. 20. Pharodoris philippinensis, new genus and species, holotype. A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 75 um. innate, very elongate branchial leaves. Ven- trally, the oral tentacles consist of two triangu- lar prolongations on each side of the buccal area. The anterior border of the foot is grooved but not notched. The mantle margin is narrower than the foot. The color of the living animal is unknown. The preserved specimen is uniformly pale cream. Anatomy The labial armature is unarmed. The radu- lar formula is 39 x (44.0.44) in a 19 mm pre- 266 VALDÉS FIG. 21. Pharodoris philippinensis, new genus and species, holotype, reproductive system. Abbrevia- tions: a, ampulla; bc, bursa copulatrix; d, deferent duct; f, female glands; pr, prostate; s, seminal re- ceptacle; sa, copulatory spine sac; sp, copulatory spine; v, vagina. served length specimen. There are no rachid- ian teeth. The innermost lateral teeth are ha- mate, having a single cusp, and they lack denticles (Fig. 20A). Some inner lateral teeth may have a small, rounded tubercle on their inner side. The teeth increase in size gradu- ally towards the medial portion of the half-row (Fig. 20B). The mid-lateral teeth are hamate, with a long cusp, and they lack denticles. The three outermost teeth are very small and have 8-10 small denticles (Fig. 20C). The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 21). The oviduct enters the female glands near the nidamental opening. The prostate is tubular and elongate; it narrows and expands again into the short deferent duct. The deferent duct opens into a common atrium with the vagina. The atrium has two large glands, each one containing a bifid, rigid spine about 1 mm long (Figs. 19A, B, 21). The penis is unarmed. The vagina is long and wide. At its proximal end, the vagina connects to the large and rounded bursa copulatrix. From the bursa copulatrix leads a short duct that branches into the elon- gate seminal receptacle and the uterine duct. Etymology From the Philippines, the type locality of this species. Remarks Pharodoris philippinensis differs from P. di- aphora in the radula and reproductive system morphology. The innermost lateral teeth of B. diaphora have a large denticle that is absent in B. philippinensis. Only few inner teeth of B. philippinensis have a small, rounded denticle on the inner side, whereas all the inner later- als of B. diaphora have a large, conical denti- cle. From the bursa copulatrix of B. philip- pinensis emerge two ducts, one connecting to the vagina and the other, very short, branch- ing into the seminal receptacle and the uterine duct, whereas in B. diaphora a single duct emerges, branching into the vagina, the sem- inal receptacle and the uterine duct. Pharodoris sp. (Fig. 22) Material Examined Banc Argo, Coral Sea, Musorstom 5 Expe- dition, stn. 290 (23°06'S, 159°26’E), 300 m, 11 October 1986, 14 mm preserved length, dissected, leg. P. Bouchet, B. Métivier and B. Richer de Forges (ММНМ). External Morphology The body is somewhat elevated, oval. The dorsum is covered with simple, conical tuber- cles. Some of them are clearly larger than the others. The branchial sheath is elevated, ris- ing clearly over the dorsal surface of the body. The perfoliate rhinophores are composed of 22 lamellae. There are six bipinnate, very elongate branchial leaves. Ventrally, the oral tentacles consist of two triangular prolonga- tions on each side of the buccal area. The an- terior border of the foot is grooved but not notched. The mantle margin is as wide as the foot. The color of the living animal is unknown. The preserved specimen is uniformly pale cream. Anatomy The labial armature is unarmed. The radu- lar formula is 33 x (40.0.40) in a 14 mm pre- served length specimen. The are no rachidian teeth. The innermost lateral teeth are hamate, having a single cusp, and they general lack denticles (Fig. 22A). Some inner lateral teeth may have a small, rounded tubercle on their inner side. The teeth increase in size gradu- DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 267 FIG. 22. Pharodoris sp., specimen from Musorstom 5 (stn. 290). A. Inner lateral teeth, scale bar = 43 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 60 um. ally towards the medial portion of the half-row (Fig. 22B). The mid-lateral teeth are hamate, with a long cusp, and they lack denticles. The three outermost teeth are very small and have 5-8 small denticles (Fig. 22C). The single specimen has the reproductive system partially developed. The two large glands associated with the atrium are visible, but some other organs are not discernible. Remarks The radula of Pharodoris sp. is very similar to that of Pharodoris philippinensis. In both, the mid-lateral teeth lack denticles, which are only present in the innermost and outermost lateral teeth. According to the available infor- mation, it is very likely that both animals be- long to the same species. However, in view of 268 VALDÉS the incomplete development of genital char- acters for this Pharodoris sp., | keep it provi- sionally separated. Moreover, since there is no information on the reproductive system and the external coloration of Pharodoris sp., | will leave it unnamed until more material and information becomes available. Genus Geitodoris Bergh, 1891 Type Species: Doris complanata Verrill, 1880, by monotypy. Geitodoris pallida Valdés, new species (Figs 2G, 16A, 23, 24) Material Examined HOLOTYPE: Norfolk Ridge, New Caledo- nia, Bathus 3 Expedition, stn. CP833 (21°14'S, 165°55’E), 441-444 m, 30 Novem- ber 1993, 12 mm preserved length, dissected, leg. P. Bouchet, B. Richer de Forges and A. Waren (MNHN). External Morphology The body is elevated and oval (Fig. 2G). The dorsum is covered with irregular tuber- cles, having a number of spicules projecting all over the surface. There are some larger tu- bercles (up to 200 um long) surrounded by smaller tubercles (Fig. 23E). The perfoliate rhinophores are composed of 22 lamellae. There are five unipinnate branchial leaves. Ventrally, the oral tentacles are short and tri- angular. The anterior border of the foot is notched and grooved (Fig. 24F). The mantle margin is about as wide as the foot. Based on the field notes, the background color of the living animal is uniformly pale cream. There are several pale brown spots in the mantle margin. The rhinophores and gill have the same color as the dorsum. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 24D), which are attached to the body wall. The small, oval, muscular buccal bulb has two additional mus- cles. The buccal bulb is ten times shorter than the oral tube. The jaws consist of numerous thin, unicuspid elements about 30 um long (Fig. 23D). The radular formula is 18 x (23.0.23) па 12 mm preserved length speci- men. There are no rachidian teeth. The inner lateral teeth are hamate, having a single cusp, and they lack denticles (Fig. 23A). The teeth increase in size gradually towards the medial portion of the half-row (Fig. 23B). The outer- most teeth are elongate plates with number of small denticles (Fig. 23C). The esophagus is long and connects directly to the intestine. The ampulla is long; it narrows into the gon- oduct, which branches into a short oviduct and the prostate (Fig. 24C). The oviduct en- ters the female glands near the center of the mass. The prostate is flattened and granular (Fig 24B); it is divided into two portions that are clearly distinguishable by their different texture and coloration. The largest portion is pale yellow and connects to the deferent duct, whereas the smallest portion is whitish and connects to the ampulla. The deferent duct is very long and narrow. It narrows and expands again into the short ejaculatory portion. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. There is an accessory gland connected to the atrium, which has several rigid structures in- side. These structures are elongate, about 150 um long (Fig 16A). The vagina is long and folded. At its proximal end, the vagina con- nects to the large and irregular bursa copula- trix. From the bursa copulatrix leads another long and convoluted duct that connects to the seminal receptacle and the uterine duct. The bursa copulatrix is about four times larger than the seminal receptacle. In the central nervous system (Fig. 24E), the cerebral and pleural ganglia are largely fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, and three pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with two nerves leading from each. The pedal and parapedal commissures are enveloped to- gether with the visceral loop along all their length. The circulatory system (Fig. 24A) consists of a heart and two blood gland situated in front and behind the central nervous system. Etymology From the Latin pallidus (pale), in reference to the color of this species. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 269 FIG. 23. Geitodoris pallida, new species, holotype. A. Inner lateral teeth; scale bar = 75 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 75 um. D. Jaw elements; scale bar = 25 um. E. Dorsal tubercles; scale bar = 150 um. Remarks Geitodoris pallida clearly belongs to the genus Geitodoris in having the following com- bination of characters: anterior border of the foot notched and grooved, dorsal tubercles simple, presence of jaws, radula lacking rachidian teeth, inner lateral teeth hamate, outermost lateral teeth elongate, plate-like, prostate flattened, penis and vagina unarmed, presence of an accessory gland. This new species is characterized by the VALDÉS 270 FIG. 24. Geitodoris pallida, new species, holotype. A. Dorsal view of the anatomy, scale bar = 1 mm. B. Re- productive system; scale bar = 0.5 mm. C. Detail of several reproductive organs; scale bar = 0.5 mm. D. An- terior portion of the digestive system; scale bar = 1 mm. E. Central nervous system; scale bar = 0.5 mm. F. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; ag, accessory gland; b, buccal bulb; bc, bursa cop- ulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; 1, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 271 presence of several solid structures, probably copulatory spines, in the accessory gland. The only described species of the genus with these structures is Geitodoris pusae (Er. Mar- cus, 1955) described from southern Brazil. According to Er. Marcus (1955), G. pusae has a denticle on the inner side of the innermost lateral teeth, whereas it is absent in G. pallida. Another difference between these species is that the outermost lateral teeth of G. pallida have a number of small denticles that appear to be absent in G. pusae. In addition, the out- ermost teeth of G. pusae are much longer than the mid-laterals, whereas in G. pallida they are shorter. Other Pacific species of Geitodoris with a proximal geographic range are G. ohshimai Baba, 1936, and G. lutea Baba, 1937, from Japan, and G. sticta Miller, 1996, from New Zealand. Geitodoris ohshimai is clearly distin- guishable from G. pallida in having denticles on the mid-lateral teeth (Baba, 1936). Geito- doris lutea has the outermost lateral teeth very elongate, longer than the mid-laterals and smooth (Baba, 1937), whereas in G. pal- lida they are shorter than the mid-laterals and bear numerous small denticles. Geitodoris sticta has short outermost lateral teeth with denticles, but in this species there are few, ir- regular denticles on the outer side of the cusp (Miller, 1996), whereas in G. pallida there are numerous, regularly arranged denticles on the inner side of the cusp. In addition, the re- productive system of G. sticta lacks an acces- sory gland that is present in G. pallida. Miller (1996) regarded the Australian spe- cies Discodoris palma Allan, 1932, and D. crawfordi Burn, 1969, as belonging to Geito- doris. According to the redescription of D. palma by Thompson (1975), this species has very elongate, smooth outer lateral teeth, which contrast with the short, denticulate outer laterals of G. pallida. The outer radular teeth of D. crawfordi are short and hamate (Burn, 1969), more similar to those of the genus Dis- codoris, and it is very likely that it belongs in this latter genus. Unfortunately, the reproduc- tive system of neither of these species has been described, so comparisons of the mor- phology of the genital organs is not possible. Genus Discodoris Bergh, 1877 Type Species: Discodoris boholiensis Bergh, 1877, by subsequent designation by O’- Donoghue (1926). Discodoris sp. (Figs. 21, 25, 26) Material Examined North of New Caledonia, Bathus 4 Expedi- tion, stn: “CP910) (18°59’5, 1163°094B), 560-608 m, 05 September 1994, 66 mm pre- served length, leg. B. Métivier and B. Richer de Forges (MNHN). External Morphology The body is elevated and oval (Fig. 21). The dorsum is covered with simple, elongate tu- bercles, all of them about 200 um long (Fig. 25E). The perfoliate rhinophores are com- posed of 35 lamellae. There are five bipinnate branchial leaves. The rhinophoral and branchial sheaths are somewhat elevated, bearing numerous tubercles. Ventrally, the oral tentacles are short and conical. The ante- rior border of the foot is notched and grooved (Fig. 26F). The mantle margin is narrower than the foot. The color of the living animals is unknown. The preserved specimen is uniformly pale cream. The rhinophores and gill have the same color as the dorsum. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 26D), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The oral tube is about twice as long as the buccal bulb. The jaws consist of numerous irregular elements, about 15 um long (Fig. 25D). The radular formula is 21 x (25.0.25) in a 66 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are ha- mate, having a single cusp, and they lack denticles (Fig. 25A). The teeth increase in size gradually towards the medial portion of the half-row (Fig. 25A, B). The outermost teeth are also hamate and lack denticles (Fig. 25C). The esophagus is very long, convoluted and connects directly to the stomach. The ampulla is very long and convoluted; it narrows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 26B). The oviduct enters the female glands near the nidamental opening. The prostate is elongate and granular; it narrows and ex- pands again into the long, wide and muscular deferent duct. The deferent duct opens into a 272 VALDÉS у ‘ DM р y y y FIG. 25. Discodoris sp. A. Inner lateral teeth; scale bar = 200 um. B. Mid-lateral teeth; scale bar = 150 um. С. Outer lateral teeth; scale bar = 150 um. D. Jaw elements; scale bar = 25 ит. E. Dorsal tubercles; scale bar = 250 um. common atrium with the vagina. The penis is unarmed. The vagina is very wide; at its prox- imal end, it connects to the large and oval bursa copulatrix. From the vagina leads an- other long duct that connects to the seminal receptacle and the very long uterine duct (Fig. 26C). The bursa copulatrix is about the same size as the seminal receptacle. In the central nervous system (Fig. 26E), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglion, and two pleural nerves lead from each pleural ganglion. The buccal gan- glia lie near the rest of the central nervous system, joined to the cerebral ganglia by two DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 273 FIG. 26. Discodoris sp. A. Dorsal view of the anatomy; scale bar = 2 mm. B. Reproductive system; scale bar = 1 mm. С. Detail of the seminal receptacle; scale bar = 1 mm. D. Anterior portion of the digestive system; scale bar = 1 mm. E. Central nervous system; scale bar = 1 mm. F. Mouth area; scale bar = 2 mm. Abbrevi- ations: a, ampulla; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve: d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and di- gestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; 0, esophagus; ot, oral tube; p, pedal nerve; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; sg, salivary gland; st, stom- ach; t, oral tentacle; v, vagina. 274 VALDÉS long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with two nerves leading from each. The pedal and parapedal commissures are enveloped together with the visceral loop along most of their length. The circulatory system (Fig. 26A) consists of a heart and two blood glands situated in front and behind the central nervous system. Remarks This species clearly belongs to the genus Discodoris as indicated by the presence of the following characters: dorsum covered with simple tubercles, presence of jaws, radula with hamate, smooth teeth, absence of ac- cessory gland and penis, and vagina un- armed. Other features of the genus Dis- codoris present in this species are foot notched and grooved, oval digitiform tenta- cles and prostate flattened. Numerous species have been assigned to the genus Discodoris, most of them from Indo-Pacific tropical waters, but also from temperate and cold regions. Discodoris sp. is the first species described from deep waters. The long vagina of this species appears to distinguish it from other members of the genus, but the lack of anatomical studies in most members of Discodoris makes difficult comparison with this new species. Consider- ing that only one specimen of this species is available and its external coloration is un- known, | prefer not to name this species until additional material becomes available. Discodoris achroma Valdés, new species (Figs. 2H, 27, 28) Material Examined HOLOTYPE: New Caledonia, Musorstom 4 Expedition, stn. CC245 (22°07'S, 167°11’E), 415-435 m, 05 October 1985, 15 mm pre- served length, leg. P. Bouchet and B. Richer de Forges (MNHN). PARATYPES: New Caledonia, Biocal Ex- pedition, stn. CP109 (22%11'S, 167°16’E), 495-515 m, 09 September 1985, one speci- men 12 mm preserved length, dissected, leg. P. Bouchet, B. Métivier and B. Richer de Forges (CASIZ 121098). New Caledonia, Mu- sorstom 4 Expedition, stn. CP242 (22°06’S, 167°10’E), 500-550 m, 03 October 1985, one specimen 11 mm preserved length, dissected, leg. P. Bouchet and B. Richer de Forges (MNHN). Norfolk Ridge, New Caledonia, Bathus 3 Expedition, stn. CP832 (23%03'S, 166°54’E), 650-659 m, 30 November 1993, one specimen 13 mm preserved length, dis- sected, leg. P. Bouchet, B. Richer de Forges and A. Waren (MNHN). ADDITIONAL MATERIAL: Coral Sea, Mu- sorstom 5 Expedition, stn. 323 (21°13'S, 157°58’E), 970 m, 14 October 1986, 15 mm preserved length, leg. P. Bouchet, B. Metivier and В. Richer de Forges (MNHN). External Morphology The body is elevated and oval (Fig. 2H). The dorsum is covered with simple, conical tubercles, all of them about 100 um long (Fig. 27E). The perfoliate rhinophores are com- posed of 33 lamellae. There are five bipinnate branchial leaves. The rhinophoral and branchial sheaths are somewhat elevated, bearing numerous tubercles. Ventrally, the oral tentacles are conical. The anterior border of the foot is notched and grooved (Fig. 28E). The mantle margin is wider than the foot. Based on the field notes, the color of the liv- ing animals is uniform translucent white. The rhinophores and gill have the same color as the dorsum. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 28C), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The oral tube is about twice as long as the buccal bulb. The jaws consist of numerous irregular elements about 50 um long (Fig. 27D). The radular formula is 15 x (18.0.18) in a 11 mm preserved length specimen and 16 x (18.0.18) in a 13 mm preserved length speci- men. There are no rachidian teeth. The inner lateral teeth are hamate, having a small cusp, and they lack denticles (Fig. 27A). The teeth increase in size gradually towards the medial portion of the half-row (Figs. 27A, B). The out- ermost teeth are also hamate and lack denti- cles (Fig. 27C). The esophagus is very long, convoluted and connects directly to the large stomach (Fig. 28A). The ampulla is very short and simple; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 28B). The oviduct enters the female glands near the DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 275 FIG. 27. Discodoris achroma, new species, paratype from Musorstom 4 (stn. CP242). A. Inner lateral teeth; scale bar = 60 um. В. Mid-lateral teeth; scale bar = 60 um. С. Outer lateral teeth; scale bar = 43 um. D. Jaw elements; scale bar = 15 um. E. Dorsal tubercles; scale bar = 100 um. center of the mass. The prostate is very elon- gate and not flattened; it narrows and ex- pands again into the short, muscular deferent duct. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. The vagina is very elongate and convoluted; at its proximal end, it connects to the rounded bursa copulatrix. From the vagina leads an- other long duct that connects to the seminal receptacle and the uterine duct. The bursa copulatrix is twice as large as the seminal re- ceptacle. In the central nervous system (Fig. 28D), the cerebral and pleural ganglia are fused and 276 VALDÉS bu © ee FIG. 28. Discodoris achroma, new species, paratype from Bathus 3 (stn. CP832). A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 0.25 mm. C. Anterior portion of the diges- tive system; scale bar = 1 mm. D. Central nervous system; scale bar = 0.5 mm. E. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; ab, abdominal ganglion; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; 0, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; plg, pleural ganglion; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 277 distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglion, and three pleural nerves lead from each pleural ganglion. The buccal gan- glia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with three nerves leading from each. The pedal and parapedal commissures are enveloped to- gether with the visceral loop along all their length. There are three abdominal ganglia on the right side of the visceral loop. The circulatory system (Fig. 28A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Ancient Greek achromos (lacking color), in reference to the translucent white appearance of this species. Remarks This species clearly belongs to the genus Discodoris by the presence of the following characters: dorsum covered with simple tu- bercles, presence of jaws, radula with ha- mate, smooth teeth, absence of accessory gland and penis, and vagina unarmed. It dif- fers from other shallow-water members of the genus in having a translucent white coloration and lacking a flattened prostate. Discodoris achroma differs from Discodoris sp. in both the external morphology and anatomy. Discodoris achroma is a small, translucent species, whereas Discodoris sp. is large and uniformly cream. Internally, they differ in the shape of the radular teeth. The in- nermost lateral teeth of D. achroma are longer and have a shorter cusp. The outermost teeth are thinner and comparatively smaller in D. achroma than in Discodoris sp. The reproduc- tive system of D. achroma has the bursa cop- ulatrix connected to the vagina by a long, con- voluted duct, whereas in Discodoris sp. this duct is much wider and shorter. The uterine duct of Discodoris sp. connects near the sem- inal receptacle, instead of near the bursa cop- ulatrix insertion, as in D. achroma. The am- pulla of D. achroma is short and simple, being very long and convoluted in Discodoris sp. The specimen from the Coral Sea was very poorly preserved and a definitive identification is not possible. The radula is very similar to that of the New Caledonia specimens of D. achroma, and it is therefore included in this species. Genus Peltodoris Bergh, 1880 Type Species: Peltodoris atromaculata Bergh, 1880, by subsequent designation by O’- Donoghue (1929). Peltodoris lippa Valdés new species (Figs. 29A, 30, 31) Material Examined HOLOTYPE: Vanuatu, Musorstom 8 Expe- dition, "sin. EP11327(15238/5, 1972037E). 161-182 m, 11 October 1994, 23 mm pre- served length, leg. P. Bouchet and B. Richer de Forges (MNHN). External Morphology The body is flat and oval (29A). The dorsum is covered with simple, conical tubercles. Some tubercles are clearly larger than others and can reach over 200 um long (Fig. 30D). The perfoliate rhinophores are composed of 16 lamellae. There are six tripinnate branchial leaves. The rhinophoral and branchial sheaths are low and bear numerous tubercles. Ven- trally, the oral tentacles are conical. The ante- rior border of the foot is notched and grooved (Fig. 31E). The mantle margin is as wide as the foot. Based on the field notes, the color of the liv- ing animal is uniform translucent white. The rhinophores and gill have the same color as the dorsum. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 31D), which are attached to the body wall. The oval, muscular buccal bulb has two large additional muscles. The oral tube is about twice as long as the buccal bulb. The labial cuticle is smooth. The radular formula is 15 x (31.0.31) па 23 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are ha- mate, having a small cusp, and they lack den- ticles (Fig. 830A). The teeth increase in size gradually towards the medial portion of the half-row (Fig. 30A, B). The outermost teeth 278 VALDÉS FIG. 29. Preserved specimens. A. Peltodoris lippa, new species, holotype; scale bar = 2 mm. B. Rostanga ankyra, new species, holotype; scale bar = 1 mm. C. Diaulula immaculata, new species, holotype; scale bar = 2 mm. D. Baptodoris phinei, new species, holotype; scale bar = 2 mm. E. Diaulula cerebralis, new species, paratype from Musorstom 3 (stn. CP128); scale bar = 3 mm. F. Dendrodoris orbicularis, new species, holo- type; scale bar = 5 mm. G. Sclerodoris virgulata new species, holotype; scale bar = 2 mm. are also hamate and have several, small den- ticles (Fig. 30C). The esophagus is very long, convoluted and connects directly to the large stomach. The ampulla is very short and simple; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 31C). The prostate is flattened, large and granular (Fig 31B); it is divided into two portions that are clearly distinguishable by their different tex- ture and coloration. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. The vagina is very elongate and convoluted; at its proximal end, it con- nects to the oval bursa copulatrix. From the bursa copulatrix leads another duct that con- nects to the seminal receptacle and the uter- ine duct. The bursa copulatrix is twice as large as the seminal receptacle. In the central nervous system (Fig. 31F), DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN FIG. 30. Peltodoris lippa, new species, holotype. A. Inner lateral teeth; scale bar = 60 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 60 um. D. Dorsal tubercles; scale bar = 150 um. the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglion, and four pleural nerves lead from each pleural ganglion. The eyes are very large and inflated. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two short nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with three (left side) and four (right side) nerves leading from them. The pedal and parapedal commissures are enveloped together with the visceral loop along all their length. There is an abdominal ganglion on the right side of the visceral loop. 280 VALDÉS FIG. 31. Peltodoris lippa, new species, holotype. A. Dorsal view of the anatomy; scale bar = 1 mm. B. Re- productive system; scale bar = 0.5 mm. C. Detail of several reproductive organs; scale bar = 0.5 mm. D. An- terior portion of the digestive system; scale bar = 0.5 mm. E. Mouth area; scale bar = 1 mm. F. Central ner- vous system; scale bar = 0.5 mm. Abbreviations: a, ampulla; b, buccal bulb; ab, abdominal ganglion; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; р, pedal nerve; pl, pleural nerve; plg, pleural ganglion; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, seminal receptacle; sg, sali- vary gland; st, stomach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 281 The circulatory system (Fig. 31A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Latin /ippus (having inflamed eyes), in reference to the large size of the eye- balls of this species. Remarks Peltodoris was regarded as a synonym of Discodoris by Thompson (1975), without any justification. Examination of specimens of Peltodoris atromaculata Bergh, 1880, the type species of the genus Peltodoris, revealed that this genus is distinguishable from Discodoris by the absence of jaw elements (Schmekel & Portmann, 1982). However, it is probable that the sole presence of this character, which is variable in other related genera, is not suffi- cient to separate Peltodoris from Discodoris. On the other hand, there may be other differ- ences between these two genera that need to be investigated. A review of the systematic status and a reconstruction of the phylogeny of the cryptobranch dorids is necessary be- fore definitive synonymies may be stabilized. In the meantime, the genus Peltodoris is here regarded as valid and different from Dis- codoris. Peltodoris lippa shares a number of fea- tures with Peltodoris atromaculata Bergh, 1880, and it clearly belongs to this genus. These features in common are dorsum with simple, non-caryophyllidia tubercles, anterior border of the foot grooved and notched, oral tentacles present, radular teeth and labial cu- ticle smooth, absence of penial hooks or ac- cessory glands, and flattened prostate. This is the first species of Peltodoris de- scribed from deep waters, and it is clearly dis- tinguishable from other species of the genus in having a uniform translucent white col- oration. The species of Peltodoris with the most proximal geographic range to P. lippa is P. noumeae (Risbec, 1937) described from shal- low waters of New Caledonia. There is not enough information in the original description of this species (Risbec, 1937) to determine whether it belongs to the genus Peltodoris. In any case, P. noumeae is clearly distinguish- able from P. lippa in having a yellow-orange background color with red spots. Peltodoris fellowsi Kay & Young, 1969, orig- inally described from Hawaii and subse- quently reported from the Pitcairn Islands (Ed- munds & Preece, 1996), has a uniform white coloration as well. However, in P fellowsi the gill is black and the rhinophores are red- brown, whereas in Р lippa rhinophores and gill are white. Internally both species differ in the radular morphology. The outermost lateral teeth of P. fellowsi are shorter than those of PR lippa. Genus Paradoris Bergh, 1884 Type Species: Paradoris granulata Bergh, 1884, by monotypy [= Paradoris indecora (Bergh, 1881)]. Paradoris araneosa Valdés, new species (165. 138, 32, 33) Material Examined HOLOTYPE: South of New Caledonia, Bathus 2 Expedition, stn. CP736 (23°03’S, 166°59’E), 452-464 m, 13 May 1993, 14 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). PARATYPES: South of New Caledonia, SMIB 8 Expedition, stn. DW182-184 (23°18’S-23°19'S, 168%05'E), 314-367 m, 31 January 1993, one specimen 35 mm pre- served length, dissected, leg. P. Bouchet and B. Richer de Forges (MNHN). Norfolk Ridge, New Caledonia, Bathus 3 Expedition, stn. DW838 (23°01'S, 166°56’E), 400-402 m, 30 November 1993, one specimen 33 mm pre- served length, dissected, leg. P. Bouchet, B. Richer de Forges and A. Waren (MNHN). Southeast of New Caledonia, Halipro 1 Expe- dition sin ©Р851 (21243/S7 166.37.E); 314-364 m, 19 March 1994, one specimen 13 mm preserved length, dissected, leg. B. Richer de Forges (CASIZ 121099). External Morphology The body is elevated and oval (Fig. 13B). The dorsum is covered with simple, rounded tubercles. The dorsal surface, in the areas without tubercles, has a number of small de- pressions surrounded by minute ridges (Fig. 32E). The perfoliate rhinophores are com- posed of 25 lamellae. There are six tripinnate branchial leaves. Ventrally, the oral tentacles 282 VALDÉS [nn —n Що FIG. 32. Paradoris araneosa, new species, paratype from Bathus 3 (stn. DW838). A. Inner lateral teeth; scale bar = 75 ит. В. Mid-lateral teeth; scale bar = 100 um. С. Outer lateral teeth; scale bar = 100 um. D. Jaw el- ements; scale bar = 43 um. E. Dorsal tubercles; scale bar = 600 um. are short and conical. The anterior border of large, dark brown spots scattered on the dor- the foot is notched and grooved (Fig. 33F). sal surface and several, more numerous, The mantle margin is about as wide as the small darker dots. The larger tubercles are foot. opaque white. The rhinophores are cream Based on the photographs, the color of the with opaque white lamellae. The branchial living animals is pale brown. There are few, leaves are pale brown and the lamellae dark DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 283 FIG. 33. Paradoris araneosa, new species, paratype from SMIB 8 (stn. DW182-184). A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 1 mm. C. Detail of the accessory organs; scale bar = 0.5 mm. D. Central nervous system; scale bar = 0.5 mm. E. Anterior portion of the digestive sys- tem; scale bar = 1 mm. F. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; ag, accessory gland; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral gan- glion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and di- gestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; 0, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, seminal re- ceptacle; sa, accessory sac; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. 284 VALDÉS brown. There are several white dots on the gill, more densely arranged near the base of the branchial leaves. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 33E), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb is about as long as the oral tube. The jaws consist of numerous thin, unicuspid elements about 30 um long (Fig. 32D). The radular formula is 68 x (16.0.16) па 32 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are ha- mate, having a single cusp, and they lack denticles (Fig. 32A). The teeth are very elon- gate with a short, strong cusp. The teeth in- crease in size gradually towards the medial portion of the half-row (Figs. 32A, B). The out- ermost teeth are elongate and also lack den- ticles (Fig. 32C). The esophagus is long and connects directly to the stomach. The ampulla is long and curved. It narrows into the gonoduct, which branches into a long oviduct and the prostate (Fig. 33B). The oviduct enters the female glands near the nidamental opening. The prostate is flattened and granular; it is divided into two portions that are clearly distinguishable by their differ- ent texture and coloration, the largest portion being pale yellow and connecting to the def- erent duct, whereas the smallest portion is or- ange and connects to the ampulla. The defer- ent duct is long and narrow; it narrows and expands again into the short ejaculatory por- tion. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. There is a large, ramified accessory gland and two small sacs, containing one rigid spine each, connected to the atrium (Fig. 33C). The vagina is very long; at its proximal end, it con- nects to the large and oval bursa copulatrix. From the bursa copulatrix leads another long duct that connects to the seminal receptacle and the very short uterine duct. The bursa copulatrix is about six times larger than the seminal receptacle. In the central nervous system (Fig. 33D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, and four pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, with three nerves lead- ing from each. The pedal and parapedal com- missures are enveloped together with the vis- ceral loop along all their length. The circulatory system (Fig. 33A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Latin araneosus (full of webs), in reference to the small ridges on the surface of this species, that resemble cobwebs. Remarks Paradoris araneosa belongs to the genus Paradoris in having the following combination of characters: anterior border of the foot notched and grooved, simple dorsal tuber- cles, reproductive system with an accessory gland and two rigid copulatory spines, jaws with numerous, simple elements, radula with strong teeth, which have a short cusp and lack denticles (Miller, 1995). Paradoris leuca Miller, 1995, from New Zealand, is the species that has been found in closest proximity to P. araneosa. It differs from P. araneosa in lacking accessory glands and copulatory sacs armed with spines (Miller, 1995). Due to the lack of these structures, it is likely that P. leuca does not belong to the genus Paradoris. Other differences are the shape of the jaw elements, which have a tri- angular cusp in P leuca and are rounded in P. araneosa. Externally, the dorsal tubercles of P. leuca are more evenly distributed and are smaller than those of P. araneosa. Paradoris tsurugensis Baba, 1986, from Japan, is distinguishable from P. araneosa in having two accessory glands and two copula- tory sacs armed with spines, instead of a sin- gle accessory gland (Baba, 1986). Also, P. tsurugensis has many more radular teeth than P. araneosa at comparable size. The radular formula of the former is 90 x (20-25.0.20-25), whereas it is 68 x (16.0.16) in the latter. Miller (1995) included Platydoris galbanus Burn, 1958, from Victoria, Australia, and Peltodoris fellowsi Kay & Young, 1969, from Hawaii, in the genus Paradoris, but both species lack jaws (Burn, 1958; Kay & Young, DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 285 1969), and therefore they must be placed in a different genus. Paradoris imperfecta Valdés, new species (Figs. 136, 34, 35) Material Examined HOLOTYPE: South of New Caledonia, Chalcal 2 Expedition, stn. CP18 (24°47'S, 168°09'E), 274 m, 27 October 1986, 13 mm preserved length, leg. P. Bouchet, B. Métivier and В. Richer de Forges (MNHN). External Morphology The body is elevated and oval (Fig. 13C). The dorsum is covered with small, conical tu- bercles. There are several larger tubercles surrounded by smaller tubercles. Many tuber- cles, especially those situated in the center of the body, are joined to the adjacent ones by short ridges. The perfoliate rhinophores are composed of 20 lamellae. There are five bip- innate branchial leaves. Ventrally, the oral tentacles are short and conical. The anterior border of the foot is notched and grooved (Fig. 34F). The mantle margin is about as wide as the foot. The color of the living animals is uniformly cream. There are few, irregular pale brown spots and numerous small dark brown dots on the dorsal surface. The dark brown spots are situated on the mantle margin. There are two areas on both sides of the dorsal hump with a high concentration of dark brown spots. The rhinophores are cream with the lamellae dark brown. The gill is uniformly cream. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 34E), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb is about as long as the oral tube. The jaws consist of numerous thin, unicuspid elements, about 10 um long (Fig. 35C). The radular formula is 46 x (13.0.13) in a 13 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are ha- mate, having a single cusp, and they lack denticles (Fig. 35A). The teeth are very elon- gate, with a short, strong cusp. The teeth in- crease in size gradually towards the medial portion of the half-row (Figs 35A-B). The out- ermost teeth are short and also lack denticles (Fig. 35C). The esophagus is long and con- nects directly to the stomach. The ampulla is short and simple; it narrows into the gonoduct, which branches into the oviduct and the prostate (Fig. 34B). The ovi- duct enters the female glands near the nida- mental opening. The prostate is long and granular; it is divided into two portions that are Clearly distinguishable by their different tex- ture and coloration. The larger portion is pale yellow and connects to the deferent duct, whereas the smaller portion is orange and connects to the ampulla. The deferent duct is short and narrow; it expands again into the long ejaculatory portion. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. There are two small sacs connected to the atrium, each containing a single rigid spine each. The vagina is very long; at its proximal end, it connects to the large, oval bursa copulatrix. From the bursa copulatrix leads another long duct that con- nects to the irregular seminal receptacle and the uterine duct (Fig 34C). The bursa copula- trix is about five times larger than the seminal receptacle. In the central nervous system (Fig. 34D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, and two pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, having two nerves leading from each one. The pedal and para- pedal commissures are enveloped together with the visceral loop along all their length. The circulatory system (Fig. 34A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Latin imperfectus (imperfect), in reference to the lack of accessory gland in this species and the irregular shape of the seminal receptacle. Remarks Paradoris imperfecta is clearly distinguish- able from P. araneosa in lacking an accessory 286 VALDÉS м FIG. 34. Paradoris imperfecta, new species, holotype. A. Dorsal view of the anatomy; scale bar = 1 mm. В. Reproductive system; scale bar = 1 mm. C. Detail of several reproductive organs; scale bar = 1 mm. D. Cen- tral nervous system; scale bar = 0.5 mm. E. Anterior portion of the digestive system; scale bar = 0.5 mm. F. Mouth area: scale bar = 1 mm. Abbreviations: a, ampulla; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; $ female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; plg, pleural ganglion; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, seminal receptacle; sa, accessory Sac, sg, Salivary gland; sp, accessory spine; st, stomach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 287 FIG. 35. Paradoris imperfecta, new species, holotype. A. Inner lateral teeth; scale bar = 43 um. B. Mid-lat- eral teeth; scale bar = 43 um. C. Outer lateral teeth; scale bar = 30 um. D. Jaw elements; scale bar = 20 um. gland. In addition, P. imperfecta has only two accessory sacs containing spines, which also distinguish this species from Paradoris leuca and P. tsurugensis. Paradoris leuca does not have accessory glands or sacs, whereas P. tsurugensis has two glands and two sacs. Anatomical differences between P. arane- osa and P. imperfecta include the outermost lateral teeth of the radula, which are much shorter in Р imperfecta. Also, the lateral teeth appear to have a longer, more triangular cusp in Р imperfecta. The prostate of P. imperfecta is more elongate than that of Р araneosa, and the seminal receptacle is irregular and elon- gate, whereas in P. araneosa it is regularly oval. Also, the atrium of P imperfecta is much 288 VALDÉS larger and the vagina is shorter than those of P. araneosa. Externally, these two species differ in the coloration of the rhinophores and the gill. The rhinophores of P. araneosa have white lamel- lae, whereas in P. imperfecta the lamellae are dark brown. The gill of P. araneosa is pale brown with dark brown lamellae and white dots, whereas it is uniformly cream in P. im- perfecta. Genus Diaulula Bergh, 1878 Type Species: Doris sandiegensis Cooper, 1863, by monotypy. Diaulula immaculata Valdés, new species (Figs. 29C, 36, 37) Material Examined HOLOTYPE: Loyalty Ridge, Musorstom 6 Expedition, stn. DW428 (20°24’S, 166°13’E), 420 m, 17 February 1989, 19 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). PARATYPE: Chesterfield, Coral Sea, Mu- sorstom 5 Expedition, stn. 371 (19°55’S, 158°38'E), 350 m, 20 October 1986, one specimen 9 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). External Morphology The body is elevated and oval (Fig. 29C). The dorsum is covered with thin caryophyl- lidia, the largest reaching over 75 um (Fig. 36D). They have a large, rounded ciliated tu- bercle and 6-7 spicules around it. The perfo- liate rhinophores are composed of 20 lamel- lae. There are four bipinnate branchial leaves. Ventrally, the oral tentacles are short and con- ical. The anterior border of the foot is notched and grooved (Fig. 37E). The mantle margin is wider than the foot. The color of the living animals is unknown. Preserved specimens are uniformly pale cream. The rhinophores and gill are also pale cream. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 37D), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The oral tube is about twice as long as the buccal bulb. The labial cuticle is smooth. The radular formula is 32 x (46.0.46) in a 9 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are hamate, hav- ing a single cusp, and they lack denticles (Fig. 36A). The teeth are very elongate with a short, strong cusp. The teeth increase in size gradu- ally towards the medial portion of the half-row (Figs. 36A, B). The outermost teeth are short and also lack denticles (Fig. 36C). The esoph- agus connects directly to the stomach. The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into the oviduct and the prostate (Fig. 37C). The oviduct enters the female glands near the center of the mass. The prostate is very large and granular (Fig 37B); it is divided into two portions that are clearly distinguishable by their different texture and coloration; the largest portion is pale yellow and connects to the deferent duct, whereas the smallest por- tion is orange and connects to the ampulla. The deferent duct is long and narrow, and opens into a common atrium with the vagina. The penis is unarmed. The vagina is long; at its proximal end, it connects to the large, oval bursa copulatrix. From the bursa copulatrix leads another long duct that connects to the seminal receptacle and the uterine duct. The bursa copulatrix is about six times larger than the seminal receptacle. The circulatory system (Fig. 37A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Latin immaculatus (unspotted), in reference to the uniformly white dorsum of this species. Remarks Valdés & Gosliner (in press) revised the type species of the genus Diaulula and pro- posed a new diagnosis of the genus that in- cludes the following characteristics: dorsum covered with elongate caryophyllidia, prostate flattened, with two portions, penis and vagina unarmed, labial cuticle smooth and radular teeth hamate and smooth. Diaulula immacu- lata has all these features and clearly belongs to this genus. Several species have been assigned to the DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 289 FIG. 36. Diaulula immaculata, new species, paratype. A. Inner lateral teeth; scale bar = 25 ит. В. Mid-lateral teeth; scale bar = 30 um. C. Outer lateral teeth; scale bar = 20 um. D. Caryophyllidia; scale bar = 75 um. genus Diaulula after its original description, which was based on Doris sandiegensis Cooper, 1863, from California. Bergh (1905) described Diaulula rubra from the Philippines, but the description is very incomplete and it is impossible to determine the correct generic placement of this species. Two years later, Bergh (1907) described two new species from South Africa, D. capensis and, with question, D. morosa. Both species appear to have caryophyllidia on the dorsum, but their repro- ductive systems are not described in enough detail to determine their correct generic place- ment. In any case, these species are distin- 290 VALDÉS [>= FIG. 37. Diaulula immaculata, new species, paratype. A. Dorsal view of the anatomy; scale bar = 1 mm. B. Reproductive system; scale bar = 0.5 mm. C. Detail of several reproductive organs; scale bar = 0.5 mm. D. Anterior portion of the digestive system; scale bar = 1 mm. E. Mouth area; scale bar = 0.5 mm. Abbrevia- tions: a, ampulla: b, buccal bulb; bc, bursa copulatrix; bg, blood gland; а, deferent duct; f, female glands; h, hermaphrodite and digestive glands; ht, heart; i, intestine; m, retractor muscle; o, esophagus; ot, oral tube; pr, prostate; s, seminal receptacle; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 291 guishable from D. immaculata by their exter- nal coloration, light brown in D. capensis, dark greenish in D. morosa, and uniformly white in D. immaculata. Eliot (1907) transferred Doris vestita Abra- ham, 1877, to the genus Diaulula. Again, this species appears to have caryophyllidia, but the internal anatomy is not described in enough detail and its generic placement re- mains uncertain. Er. Marcus (1959) re- described Doris hispida d'Orbigny, 1837, and placed this species in the genus Diaulula. This species has a dorsal ridge and could be a member of the genus Atagema J. E. Gray, 1850 (Valdés & Gosliner, in press). Diaulula immaculata is clearly distinguish- able from D. sandiegensis by both the exter- nal morphology and anatomy. Externally, D. sandiegensis has very distinctive black circles or spots on the dorsum that are absent in D. immaculata. Also, the outermost radular teeth of D. sandiegensis are more elongated and the prostate comparatively smaller than those of D. immaculata. Diaulula cerebralis Valdés, new species (Figs 29E, 38, 39) Material Examined HOLOTYPE: Philippines, Musorstom 3 Ex- pedition, stn. CP144 (11°02’N, 124°15’E), 379-383 m, 7 June 1985, 30 mm preserved length, leg. P. Bouchet and M. P. Triclot (MNHN). PARATYPE: Philippines, Musorstom 3 Ex- pedition, stn. CP128 (11°50’М, 121°42’E), 815-321 m, 5 June 1985, one specimen 27 mm preserved length, dissected, leg. P. Bouchet and M. P. Triclot (MNHN). External Morphology The body is elevated and oval (Fig. 29E). The dorsum is covered with long, thin caryophyllidia, the largest reaching over 250 um (Fig. 38E). They have a large, oval ciliated tubercle and four long spicules around it. The perfoliate rhinophores are composed of 30 lamellae. There are eight bipinnate branchial leaves. The rhinophoral and branchial sheaths are elevated. Ventrally, the oral tenta- cles are short and conical. The anterior border of the foot is notched and grooved (Fig. 39F). The mantle margin is about as wide as the foot. The color of the living animals is uniformly white. The rhinophores and gill are also white. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 39E), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb is about as long as the thin oral tube. The jaws consist of numerous thin, uni- cuspid elements, about 10 um long (Fig. 38D). The radular formula is 21 x (21.0.21) in a 27 mm preserved length specimen. There are no rachidian teeth. The three innermost lateral teeth are simple plates, lacking a cusp and denticles (Fig. 38A). The following lateral teeth have a single, short cusp and also lack denticles (Figs 38A-B). The teeth increase in size gradually towards the medial portion of the half-row. The outermost teeth are short, hamate and also lack denticles (Fig. 38C). The esophagus is long and connects directly to the stomach (Fig. 39A). The ampulla is long and very convoluted; it narrows into the gonoduct, which branches into the short oviduct and the prostate (Fig. 39C). The oviduct enters the female glands near the nidamental opening. The prostate is long and granular (Fig. 39B); it narrows and expands again into the very large deferent duct, which opens into a common atrium with the vagina. The penis is unarmed. The vagina is wide and long; at its proximal end, it con- nects to the oval bursa copulatrix. From the distal end of the vagina leads another long duct that connects to the seminal receptacle and the uterine duct. The bursa copulatrix is about two times larger than the seminal re- ceptacle. In the central nervous system (Fig. 39D), the cerebral and pleural ganglia are partially fused and distinct from the pedal ganglia. The cerebral ganglia are very large and conspicu- ous. There are two cerebral nerves leading from each cerebral ganglion, and three (right side) or two (left side) pleural nerves lead from the pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral and optical ganglia are present. The pedal ganglia are clearly separated, having three (right side) or four (left side) pedal nerves leading from them. The pedal and parapedal commissures are enveloped together with the visceral loop along all their length. 292 VALDÉS FIG. 38. Diaulula cerebralis, new species, paratype. A. Inner lateral teeth; scale bar = 100 um. B. Mid-lateral teeth; scale bar = 100 um. C. Outer lateral teeth; scale bar = 100 um. D. Jaw elements; scale bar = 20 um. E. Caryophyllidia; scale bar = 250 um. The circulatory system (Fig. 39A) consists of a heart and two blood glands situated in front and behind the central nervous system. Etymology From the Latin cerebrum (brain), in refer- ence to the extremely developed cerebral ganglia of this species. Remarks The generic position of this species is par- ticularly problematic. It fits within the genus Diaulula in having the dorsum covered with elongate caryophyllidia, prostate flattened, with two portions, penis and vagina unarmed and radular teeth hamate and smooth. How- ever, it is different from the other species of DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 293 FIG. 39. Diaulula cerebralis, new species, paratype. A. Dorsal view of the anatomy; scale bar = 1 mm. B. Re- productive system; scale bar = 1 mm. C. Detail of several reproductive organs; scale bar = 1 mm. D. Central nervous system; scale bar = 0.5 mm. E. Anterior portion of the digestive system; scale bar = 0.5 mm. F. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, re- tractor muscle; 0, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. 294 VALDÉS this genus in having jaws. The presence or absence of jaws is variable in other genera, such as Rostanga and Jorunna, and may not have much phylogenetic information (Valdés & Gosliner, in press). Other related genera in which this species could fit are Rostanga and Jorunna. However, Rostanga has very elon- gate and denticulate lateral teeth, and Jorunna has a large accessory gland with a copulatory spine. These characters are ab- sent in Diaulula cerebralis. Therefore, this species is provisionally placed in the genus Diaulula where it is better accommodated. Diaulula cerebralis differs from D. immacu- lata in having a much smaller prostate, the seminal receptacle connected to the vagina, instead of to the bursa copulatrix, the inner- most lateral teeth like simple plates, and the cerebral ganglia extremely developed com- pared to other dorid central nervous systems previously studied (Valdés & Gosliner, in press). However, the most important differ- ence between these two species is the pres- ence of jaws in Diaulula cerebralis. Diaulula sandiegensis is also different from D. cere- bralis in having dorsal black circles or spots and lacking jaws. Genus Rostanga Bergh, 1879 Type Species: Doris coccinea Forbes, in Alder and Hancock, 1848, by original designa- tion [= Rostanga rubra (Risso, 1818)]. Rostanga ankyra Valdes, new species (Figs. 29B, 40, 41) Material Examined HOLOTYPE: South of New Caledonia, Спаса! 2 Expedition, stn. DW74 (24°40'S, 168”38'E), 650 m, 29 October 1985, 8 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). External Morphology The body is elevated and oval (Fig. 29B). The dorsum is covered with long, thin caryophyllidia, the largest reaching over 100 um (Fig. 40D). They have a large ciliated tu- bercle and 5-6 spicules around it. The perfo- liate rhinophores are composed of 19, trans- verse lamellae. There are five bipinnate branchial leaves. Ventrally, the oral tentacles are short and conical. The anterior border of the foot is notched and grooved. The mantle margin is about as wide as the foot. The color of the living animal is uniformly white. The rhinophores and gill are also white. Anatomy The single specimen was poorly preserved and only the radula and reproductive system were examined. The labial cuticle is smooth. The radular formula is 77 x (97.0.97) in a 8 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are ha- mate, having a long, thin cusp folded inwards, and a single, long denticle on the outer side (Fig. 40A). The base is very wide, triangular. The teeth increase in size gradually towards the medial portion of the half-row (Fig. 40A, B). The mid-lateral teeth are very long, with a Curved cusp and two long denticles. The out- ermost teeth are also very elongate, with a single, long denticle (Fig. 40C). The ampulla is short; it narrows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 41). The oviduct enters the female glands near the center of the mass. The prostate is long and granular; it is divided into two portions that are clearly dis- tinguishable by their different texture and col- oration. The deferent duct is very long and narrow; it opens into a common atrium with the vagina. The vagina is very long; it expands into a large, muscular portion that narrows again before it connects to the large and ir- regular bursa copulatrix. From the bursa cop- ulatrix leads another long duct that connects to the seminal receptacle and the short uter- ine duct. The bursa copulatrix is about twenty times larger than the seminal receptacle. Etymology From the Ancient Greek ankyra (anchor), in reference to the shape of the innermost radu- lar teeth of this species. Remarks The radular morphology of R. ankyra is typ- ical of the genus Rostanga. It is characterized by having the innermost teeth with a triangu- lar, wide base and a cusp folded inwards, and the outermost teeth very elongate. Also, the oval body shape and the presence of long caryophyllidia on the dorsum clearly suggest DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 295 FIG. 40. Rostanga ankyra, new species, holotype. A. Inner lateral teeth; scale bar = 15 um. B. Mid-lateral teeth; scale bar = 25 um. С. Outer lateral teeth; scale bar = 20 ит. D, Caryophyllidium; scale bar = 43 um. the placement of this species in the genus Rostanga. Rudman & Avern (1989) revised the genus Rostanga in the Indo-West Pacific. According to these authors there are nine valid species in this region, all of them characterized by a red or orange general color. More recently, Garovoy et al. (in press) described three new species from South Africa, and included Bore- odoris setidens Odhner, 1939, in the genus Rostanga. Rostanga setidens, as well as one of the new species from South Africa, have a white background color. However, the new species from South Africa is clearly distin- guishable from R. ankyrain having the dorsum covered with purplish brown spots (Garovoy et 296 VALDÉS FIG. 41. Rostanga ankyra, new species, holotype. Reproductive system. Abbreviations: a, ampulla; bc, bursa copulatrix; d, deferent duct; f, female glands; pr, prostate; s, seminal receptacle; v, vagina. al., in press). Other differences include the presence of jaw elements, and the lack of den- ticles on the mid-lateral teeth of the new species from South Africa. Rostanga setidens, on the other hand, has a radular morphology very similar to R. ankyra, and both species lack jaws. The main radular difference is the presence of a single denticle in all the lateral teeth of R. setidens (see Odhner, 1939), whereas in the mid-lateral teeth of R. ankyra there are two long denticles. The reproductive system of A. ankyra has a very long deferent duct, and a wide, muscular portion in the vagina, whereas in R. setidens the deferent duct is shorter, and the vagina is simple. Genus Sclerodoris Eliot, 1904 Type Species: Sclerodoris tuberculata Eliot, 1904, by subsequent designation herein. Sclerodoris virgulata Valdés, new species (Figs. 29G, 42, 43) Material Examined HOLOTYPE: South of New Caledonia, Chalcal 2 Expedition, stn. CP27 (23%15'S, 168°05’E), 283 m, 31 October 1985, 14 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). PARATYPES: South of New Caledonia, Chalcal 2 Expedition, stn. CP27 (23%15'S, 168°05'E), 283 m, 31 October 1985, one specimen 12 mm preserved length, leg. P. Bouchet, B. Metivier and B. Richer de Forges (CASIZ 121100). Norfolk Ridge, New Caledo- nia, Bathus 3 Expedition, stn. CP847 (23°03'$, 166°58'E), 405-411 m, 1 Decem- ber 1993, one specimen 10 mm preserved length, leg. P. Bouchet, B. Richer de Forges and A. Waren (MNHN). External Morphology The body is elevated and oval (Fig. 29G). The dorsum is covered with long, thin caryo- phyllidia, the largest reaching over 100 um (Fig. 42D). They have a small ciliated tubercle and 4-5 spicules around it. There are no dor- sal ridges or depressions. The perfoliate rhinophores are composed of 21 lamellae. There are four long, thin, bipinnate branchial leaves. Ventrally, the oral tentacles are short and conical (Fig. 43E). The anterior border of the foot is notched and grooved. The mantle margin is about as wide as the foot. The color of the living animals is white, with a series of dark brown spots and lines. These spots and lines are arranged in a regular pat- tern. There are numerous spots in the center of the dorsum that in two specimens may line up to form a line, running from the rhino- phores to the gill. Along the mantle margin there are several lines projecting radially from the center of the dorsum. There are small spots distributed irregularly among the central and radial lines. The rhinophores and gill are also white. Anatomy The posterior end of the oral tube has six strong retractor muscles (Fig. 43C), which are attached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb is about as long as the oral tube. The labial cuticle is smooth. The radular for- mula is 31 x (41.0.41) in a 10 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are hamate, having a single cusp, and they lack denticles (Fig. 42A). The teeth increase in size suddenly towards the medial portion of the half-row. The mid-lat- eral teeth are also hamate and lack denticles DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 297 FIG. 42. Sclerodoris virgulata, new species, paratype from Bathus 3 (stn. CP847). A. Inner lateral teeth; scale bar = 25 um. B. Mid-lateral teeth; scale bar = 43 um. C. Outer lateral teeth; scale bar = 30 um. D. Caryophyl- lidium; scale bar = 30 um. E. Penial hooks; scale bar = 43 um. (Fig. 42B). The outermost teeth are elongate, with a short base, and have 3-11 long denti- cles (Fig. 42C). The esophagus is long and connects directly to the stomach (Fig. 43A). The ampulla is long; it narrows into the gon- oduct, which branches into a short oviduct and the prostate (Fig. 43B). The oviduct en- ters the female glands near the nidamental opening. The prostate is flattened and granu- lar; it is divided into two portions that are clearly distinguishable by their different tex- ture and coloration. The deferent duct is long and narrow; it opens into a common, large atrium with the vagina. The penis is armed 298 VALDÉS — FIG. 43. Sclerodoris virgulata, new species, paratype from Bathus 3 (stn. CP847). A. Dorsal view of the anatomy; scale bar = 0.5 mm. B. Reproductive system; scale bar = 0.25 mm. C. Anterior portion of the di- gestive system; scale bar = 0.5 mm. D. Central nervous system; scale bar = 0.25 mm. E. Mouth area; scale bar = 0.5 mm. Abbreviations: a, ampulla; ab, abdominal ganglion; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; gr, glandular region of the retractor muscles; h, hermaphrodite and digestive glands; ht, heart; i, intestine; |, visceral loop; m, retractor muscle; o, esophagus; ot, oral tube; p, pedal nerve; pg, pedal ganglion; pl, pleural nerve; plg, pleural ganglion; pr, prostate; r, rhinophoral nerve; s, seminal receptacle; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 299 with large, triangular hooks, about 70 um long (Fig. 42E). The vagina is very long; at its prox- imal end, it connects to the large and irregular bursa copulatrix. From the bursa copulatrix leads another long duct that connects to the seminal receptacle and the very short uterine duct. The bursa copulatrix is about ten times larger than the seminal receptacle. In the central nervous system (Fig. 43D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, and two (left) or one (right) pleural nerves lead from the pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral, and optical ganglia are present. The pedal ganglia are clearly separated, hav- ing three nerves leading from each one. The pedal and parapedal commissures are en- veloped together with the visceral loop along all their length. There is an abdominal ganglia on the right side of the visceral loop. The circulatory system (Fig. 43A) consists of a heart and two blood gland situated in front and behind the central nervous system. Etymology From the Latin virgulatus (striped), in refer- ence to the numerous black lines on the dor- sum of this species. Remarks Rudman (1978) revised the genus Scle- rodoris in the Indo-West Pacific. He recog- nized eight valid species in this area (two of them unnamed), all of them very different from Sclerodoris virgulata. All the species previ- ously described have either dorsal ridges or depressions, which are absent in S. virgulata. Moreover, the combination of the following characters makes S. virgulata very easily dis- tinguishable from other members of the genus: white color pattern with radial dark brown lines, absence of an accessory gland, caryophyllidia lacking a projecting filament, and presence of four, thin branchial leaves. Another genus with caryophyllidia and den- ticulated outermost teeth is Taringa (Valdés & Gosliner, in press). However, S. virgulata does not fit in Taringa in having penial hooks and lacking a cuticular structure in the penis. In addition, Taringa has the inner and mid-lat- eral teeth denticulated, whereas they are smooth in S. virgulata. Genus Baptodoris Bergh, 1884 Type Species: Baptodoris cinnabarina Bergh, 1884, by monotypy. Baptodoris phinei Valdés, new species (Figs. 29D, 44, 45) Material Examined HOLOTYPE: Coral Sea, Musorstom 5 Ex- pedition, stn. 324 (21%15'S, 157°51’E), 970 m, 21 mm preserved length, leg. P. Bouchet, B. Métivier and B. Richer de Forges (MNHN). External Morphology The body is elevated and oval (Fig. 29D). The dorsum is covered with conical caryophyllidia, the largest reaching over 100 um (Fig. 44D). They have a large, elongate ciliated tubercle and five spicules around it. The perfoliate rhinophores are composed of 32 lamellae. There are five tripinnate branchial leaves. The rhinophoral and branchial sheaths are low. Ventrally, the oral tentacles are short and conical. The anterior border of the foot is notched and grooved (Fig. 45E). The mantle margin is narrower than the foot. The color of the living animal is uniformly white. The rhinophores and gill are also white. Anatomy The posterior end of the oral tube has six retractor muscles (Fig. 45C), which are at- tached to the body wall. The oval, muscular buccal bulb has two additional muscles. The buccal bulb is about as long as the oral tube. The labial cuticle is smooth. The radular for- mula is 20 x (18.0.18) in a21 mm preserved length specimen. There are no rachidian teeth. The inner lateral teeth are hamate, hav- ing a single cusp with a large, rounded denti- cle (Fig. 44A). The teeth are very elongate with a short, strong cusp. The teeth increase in size gradually towards the medial portion of the half-row (Fig. 44A, B). The outermost teeth are short and have several, very small denticles (Fig. 44C). The esophagus is very long and wide, and connects directly to the stomach. 300 VALDÉS FIG. 44. Baptodoris phinei, new species, holotype. A. Inner lateral teeth; scale bar = 43 um. B. Mid-lateral teeth; scale bar = 60 um. C. Outer lateral teeth; scale bar = 75 um. D. Caryophyllidia; scale bar = 150 um. The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into the short oviduct and the prostate (Fig. 45B). The oviduct enters the female glands near the nidamental opening. The prostate is large and granular; it is divided into two portions that are clearly distinguishable by their different tex- ture and coloration; the largest portion is pale yellow and connects to the deferent duct, whereas the smallest portion is orange and connects to the ampulla. The deferent duct is long and narrow; it expands into the ejacula- tory portion, which opens into a common atrium with the vagina. The penis is unarmed. There is a simple accessory gland that opens into the atrium, between the deferent duct and DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 301 E FIG. 45. Baptodoris phinei, new species, holotype. A. Dorsal view of the anatomy; scale bar = 1 mm. B. Re- productive system; scale bar = 1 mm. C. Anterior portion of the digestive system; scale bar = 1 mm. D. Cen- tral nervous system; scale bar = 0.5 mm. E. Mouth area; scale bar = 1 mm. Abbreviations: a, ampulla; ab, abdominal ganglion; ag, accessory gland; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; d, deferent duct; e, esophageal ganglion; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; 1, visceral loop; m, retractor muscle; 0, esopha- gus; ot, oral tube; p, pedal nerve; pl, pleural nerve; pr, prostate; r, rhinophoral nerve; rs, renal syrinx; s, sem- inal receptacle; sg, salivary gland; st, stomach; t, oral tentacle; v, vagina. 302 VALDÉS the vagina. The vagina is short; at its proximal end, it connects to the large, oval bursa copu- latrix. From the distal portion of the vagina leads another long duct that connects to the seminal receptacle and the uterine duct. The bursa copulatrix is about twice as large as the seminal receptacle. In the central nervous system (Fig. 45D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are three cerebral nerves leading from each cere- bral ganglion, and three pleural nerves lead from each pleural ganglion. The buccal gan- glia lie near the rest of the central nervous system, joined to the cerebral ganglia by two long nerves. Gastroesophageal, rhinophoral and optical ganglia are present. There are no eyes. The pedal ganglia are clearly sepa- rated, having three nerves leading from each. A large abdominal ganglion lies on the right side of the central nervous system. The pedal and parapedal commissures are enveloped together with the visceral loop along all their length. The circulatory system (Fig. 45A) consists of a heart and a single blood gland situated over the central nervous system. Etymology Dedicated to Phineus, the mythical prophet who was blinded by Zeus, in reference to the lack of eyes of this species. Remarks The genus Baptodoris was recently diag- nosed by Valdés & Gosliner (in press) as hav- ing the dorsum covered with caryophyllidia, prostate with two portions, penis armed with hooks, and a gap without hooks near the opening, accessory gland lobate, without a spine, labial cuticle smooth, and outermost lateral teeth very small and multidenticulate. Baptodoris phinei has all these characteristics except for the penial hooks. However, B. phinei share more features with members of the Baptodoris clade than with other caryoph- yllidia-bearing dorids and therefore it is provi- sionally placed in this genus. Also, B. phinei share with one of the two valid species of the genus, Baptodoris cinnabarina Bergh, 1884, the presence of a single, large denticle on the cusp of the lateral radular teeth. Baptodoris phinei is clearly distinguishable from the two other members of the genus Baptodoris by the external morphology and anatomy. Baptodoris cinnabarina is yellow to dark red with white and brown spots (Valdés & Gosliner, in press), and B. mimetica Gosliner, 1991, is bright lemon yellow with white spots (Gosliner, 1991), whereas P. phinei is uni- formly pale cream. The anatomical unique features of P. phinei are the lack of eyes and the absence of penial hooks. Genus Dendrodoris Ehrenberg, 1831 Type Species: Dendrodoris lugubris Ehren- berg, 1831, by subsequent designation by Gray (1847). Dendrodoris orbicularis Valdés, new species (Figs. 29F, 46) Material Examined HOLOTYPE: Southeast of New Caledonia, Halipro 1 Expedition, stn. CC856 (21°44'S, 166°37'E), 311-365 m, 20 March 1994, 47 mm preserved length, dissected, leg. B. Richer de Forges (MNHN). PARATYPES: New Caledonia, Musorstom 4 Expedition, stn. CP135 (18555, 163°22'E), 456 m, 19 September 1985, one specimen 46 mm preserved length, dis- sected, leg. P. Bouchet and B. Richer de Forges (MNHN). Southeast of New Caledo- nia, Halipro 1 Expedition, stn. CC851 (21°43'S, 166°37'E), 314-364 m, 19 March 1994, one specimen 14 mm preserved length, dissected, leg. B. Richer de Forges (MNHN). North of New Caledonia, Bathus 4 Expedition, stn. CP906 (19°01'S, 163°15’E), 339-350 m, 4 September 1994, one specimen 20 mm pre- served length, dissected, leg. B. Métivier and B. Richer de Forges (CASIZ 121101); stn. DW925 (18°55'S, 163°24’E), 370-405 m, 7 August 1994, one specimen 11 mm preserved length, dissected, leg. B. Métivier and B. Richer de Forges (MNHN). External Morphology The body is elevated, oval (Fig. 29F), soft in texture. The dorsum is smooth, lacking tuber- cles. The perfoliate rhinophores consist of 37 lamellae. There are six tripinnate branchial leaves. Ventrally there are no oral tentacles. The anterior border of the foot has a deep notch where the mouth opens (Fig. 46E). The DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 303 FIG. 46. Dendrodoris orbicularis, new species, paratype from Musorstom 4 (stn. CP135). A. Dorsal view of the anatomy; scale bar = 2 mm. B. Reproductive system; scale bar = 2 mm. C. Detail of several reproduc- tive organs; scale bar = 2 mm. D. Central nervous system; scale bar = 1 mm. E. Mouth area; scale bar = 3 mm. Abbreviations: a, ampulla; b, buccal bulb; bc, bursa copulatrix; bg, blood gland; bu, buccal ganglion; c, cerebral nerve; cg, cerebral ganglion; d, deferent duct; f, female glands; g, genital nerve; h, hermaphrodite and digestive glands; ht, heart; i, intestine; o, esophagus; p, pedal nerve; pl, pleural nerve; plg, pleural gan- glion; pr, prostate; pt, ptyaline gland; py, pyloric sac; r, rhinophoral nerve; s, seminal receptacle; v, vagina. foot is very wide, almost ten times wider than the mantle margin The color of the living animals is unknown. Only one specimen, the holotype, retained some of the original coloration. It is pale brown with several, large rings of dark brown. The interior of some ridges is darker than the rest of the dorsum. The mantle margin is edged by a thin, white line, that is interiorly surrounded by a wider black band. Ventrally, there are faded brown spots densely distrib- uted on the foot sole, and a few, larger scat- 304 VALDÉS tered on the mantle margin. The other speci- mens are uniformly cream, some of them with faded patches of dark brown. Anatomy The posterior end of the buccal bulb has four thin retractor muscles (Fig. 46A), which attach to the body wall. There are no salivary glands. Two large ptyaline glands are joined together to a single duct that opens into the buccal bulb. There are no jaws and radula. The esophagus is long and convoluted. It has two rounded esophageal glands and con- nects directly to the digestive gland. The in- testine is short and has a pyloric sac on its proximal region (Fig. 464). The ampulla is long and convoluted; it nar- rows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 46C). The oviduct enters the female glands near the center of the mass. The prostate is tubular, granular, and elongate (Fig. 46B); it narrows and expands again into the long deferent duct. The deferent duct opens into a common atrium with the vagina. The penis is unarmed. The vagina is long and wide; at its proximal end, it connects to the large, irregular bursa copulatrix. Another very long, convoluted duct, which connects to the seminal recepta- cle and to the short uterine duct, leads from the bursa copulatrix. The bursa copulatrix is about twenty times larger than the seminal re- ceptacle. In the central nervous system (Fig. 46D), the cerebral and pleural ganglia are fused and distinct from the pedal ganglia. There are four cerebral nerves leading from each cerebral ganglion, and three pleural nerves lead from each pleural ganglion. The buccal ganglia lie near the rest of the central nervous system, joined to the cerebral ganglia by two short nerves. Optical ganglia are present. The rhinophoral ganglia appear to be fused with the cerebral ganglia. The two pedal ganglia are joined together ventrally, having three nerves leading from each one. There are no pedal and parapedal commissures. The circulatory system (Fig. 46A) consists of a heart and a single blood gland situated behind the central nervous system. Etymology From the Latin orbis (circle, ring), in refer- ence to the brown rings on the dorsum of this species. Remarks Dendrodoris orbicularis is easily distin- guishable from other Indo-Pacific species of Dendrodoris by its external coloration. There are no other species described from this area with a color pattern of large, dark dorsal rings. In addition, the lack of penial hooks in D. or- bicularis differentiates this species from most of the others studied anatomically. Only Den- drodoris coronata Kay & Young, 1969, also lacks penial hooks too (Kay & Young, 1969; Valdes & Gosliner, 1999), but this species is easily distinguished from D. orbicularis in hav- ing a white coloration with black spots, dorsal tubercles, the seminal receptacle larger than the bursa copulatrix and a very short ampulla (Valdes & Gosliner, 1999). Dendrodoris brodieae Valdés, new species (Figs. 13D, 47A, B, 48A) Material Examined HOLOTYPE: North of New Caledonia, Mu- sorstom 4 Expedition, stn. CP153 (19°04'S, 163°21’E), 235 m, 14 September 1985, one specimen 14 mm preserved length, leg. P. Bouchet and B. Richer de Forges (MNHN). PARATYPES: South of New Caledonia, Musorstom 4 Expedition, stn. DW227 (22°46'S, 167°20'E), 300 m, 30 September 1985, one specimen 19 mm preserved length, leg. P. Bouchet and B. Richer de Forges (CASIZ 121102). South of New Caledonia, Chalcal 2 Expedition, stn. DW81 (23°20'S, 168°03’E), 311 m, 31 October 1986, one specimen 31 mm preserved length, dis- sected, leg. P. Bouchet, B. Métivier, B. Richer de Forges (MNHN). External Morphology The body of the living animal is elevated, oval, soft in texture (Fig. 13D). The dorsum is smooth, lacking tubercles. The perfoliate rhinophores consist of 32 lamellae. There are six tripinnate branchial leaves. Ventrally there are no oral tentacles. The anterior border of the foot has a deep notch where the mouth opens. The foot is very wide, almost ten times wider than the mantle margin. The general color of the body is pale brown. The dorsum is covered with numerous, rounded or oval pale cream spots. Among them, there are several large, dark brown patches. In some cases, the brown patches DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 305 FIG. 47. Reproductive systems. A, B. Dendrodoris brodieae, specimen from Chalcal 2 (DW81); scale bar = 1 mm. C, D. Dendrodoris sp., specimen from Musorstom 3 (stn. CP110); scale bar = 1 mm. Abbreviations: a, ampulla; bc, bursa copulatrix; d, deferent duct; f, female glands; pr, prostate; s, seminal receptacle; v, vagina; vg, vestibular gland. overlap the white spots. The mantle edge is surrounded by a white or pale cream line, in- terrupted by dark brown, almost black spots. These spots are connected to pale brown spots situated on the mantle margin. The rhinophores are cream with the apex black. The gill is also cream with the lamellae edged with dark brown pigment. Anatomy The reproductive system has a wide, sim- ple ampulla; it narrows into the gonoduct, which branches into a long oviduct and the prostate (Fig. 47B). The oviduct enters the fe- male glands near the center of the mass. The prostate is tubular, granular and elongate (Fig. 47A); it narrows and expands again into the deferent duct. The deferent duct opens into a common atrium with the vagina. The penis is armed with several rows of elongate hooks. The hooks have long bases and coni- cal cusps, about 50 um long (Fig. 48A). The vagina is long and wide; at its proximal end, it connects to the large and rounded bursa cop- ulatrix. Another long, convoluted duct, which connects to the seminal receptacle and to the short uterine duct, leads from the bursa copu- latrix. The bursa copulatrix is about ten times larger than the seminal receptacle. 306 VALDÉS FIG. 48. Penial hooks. A. Dendrodoris brodieae, specimen from Chalcal 2 (DW81); scale bar = 25 um. B. Dendrodoris sp., specimen from Musorstom 3 (stn. CP110); scale bar = 20 um. Etymology Dedicated to Gilianne Brodie, who is re- viewing the genus Dendrodoris in the Indo- West Pacific Remarks Dendrodoris brodieae is clearly distinguish- able from other Indo-Pacific species of Den- drodoris by its external coloration. The only other species with cream rounded spots on the dorsum is Dendrodoris guttata (Odhner, 1917). However, D. guttata has also black spots in the middle of the cream spots (Willan & Coleman, 1984), which are absent in D. brodieae. In addition, D. guttata has a fawn or apricot general color, whereas D. brodieae is pale brown. Also, the dark brown spots of D. brodieae are absent in D. guttata (Willan & Coleman, 1984). Dendrodoris denisoni (Angas, 1864) also resembles D. brodieae in the external coloration, but these two species are clearly distinguishable by the presence of large tubercles in the former. Internally, D. brodieae is characterized by having a very large vestibular gland. Other species of Dendrodoris with vestibular glands are D. nigra (Stimpson, 1855), D. denisoni (Angas, 1864), and D. albobrunnea Allan, 1933 (Brodie et al., 1997; Valdes & Gosliner, 1999). In D. nigra and D. denisoni, the vestibular gland is much smaller than that of D. brodieae. Only in D. albobrunnea it has a very large vestibular gland, comparable to that of D. brodieae. However, both species are distinguishable by the external morphol- ogy. Dendrodoris albobrunnea has small dor- sal tubercles, whereas D. brodieae has a smooth dorsum. Moreover, D. albobrunnea is a very pale species with small brown spots, whereas D. brodieae has a much more com- plex coloration, with brown patches and pale cream spots. Dendrodoris sp. (Figs. 47C, D, 48B) Material Examined Philippines, Musorstom 3 Expedition, stn. CP110 (14°00’N, 120°18’E), 187-193 m, 2 June 1985, one specimen 27 mm preserved length, dissected, leg. P. Bouchet and M. P. Triclot (MNHN). External Morphology The body is elevated, oval, soft in texture. The dorsum is smooth, lacking tubercles. The perfoliate rhinophores consist of 31 lamellae. There are six tripinnate branchial leaves. Ven- trally there are no oral tentacles. The anterior DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 307 border of the foot has a deep notch where the mouth opens. The foot is very wide, almost ten times wider than the mantle margin. The color of the living animal is unknown. Anatomy The reproductive system has a small, con- voluted ampulla; it narrows into the gonoduct, which branches into a short oviduct and the prostate (Fig. 47D). The oviduct enters the fe- male glands near the center of the mass. The prostate is tubular, granular and elongate (Fig. 47C); it narrows and expands again into the long and wide deferent duct. The deferent duct opens into a common atrium with the vagina. The penis is armed with several rows of penial hooks. The hooks have long bases and minute, rounded cusps, about 2 um long (Fig. 48B). The vagina is long and wide; at its proximal end, it connects to the large and rounded bursa copulatrix. Another very long and convoluted duct, which connects to the seminal receptacle and to the short uterine duct, leads from the bursa copulatrix. The bursa copulatrix is about 20 times larger than the seminal receptacle. Remarks The penial hooks of this species appear to be very distinctive. However, a single speci- men of this species was collected, and there is not information on the external coloration of the living animal. Therefore, | prefer not to name this likely new species before more ma- terial becomes available. DISCUSSION The deep-water species of cryptobranch dorids described in this paper have been compared with their known closest relatives inhabiting shallow waters. The anatomy and the generic placement of many shallow-water species of cryptobranch dorids from the tropi- cal Indo-Pacific are poorly known. It may be possible that some of the tropical shallow- water species have large bathymetric ranges, showing a pale or white coloration when they occur in deep waters. For example, the no- taspidean opisthobranch Umbraculum um- braculum Lightfoot, 1786, is found in eastern Australia from the intertidal zone to 274 m, and the deep-water specimens are translu- cent white, apparently lacking pigment (Rud- man, 1999). However, most of the species de- scribed in the present paper, closest relatives of which have been anatomically studied, show notable anatomical differences from the shallow-water species and they clearly consti- tute new species. Moreover, the evidence in- dicates that most species of cryptobranch dorids have, in general, narrow bathymetric ranges. As for the phyllidiids on the Norfolk Ridge (Valdés, 2001), the present material shows that several deep-sea assemblages replace each other in different depth zones. For instance, of the ten species found in the 200-400-m depth interval, one (Cadlina abyssicola) has been recorded from deeper than 500 m and another (Pharodoris di- aphora) from more shallow than 100 m. Con- versely, of the seven species found in the 500-1,000-m interval, only two (Cadlina abyssicola and Discodoris achroma) are recorded from shallower depths than 500 m. Even if only species represented by more than one lot are considered, there is evidence for a considerable species turnover: of the five species found at more than one station in the 200-400-m depth interval, one (Cadlina abyssicola) has been recorded from deeper than 500 m and another (Pharodoris di- aphora) from more shallow than 100 m. Con- versely, of the five species found at more than one station in the 500-1000-m interval, two (Cadlina abyssicola and Discodoris achroma) have been recorded from more shallow than 500 m. The species studied in this paper belong to 13 genera of cryptobranch dorids. Most of these genera-Cadlina, Geitodoris, Dis- codoris, Peltodoris, Paradoris, Diaulula, Ros- tanga, Sclerodoris and Dendrodoris— are here reported from deep waters for the first time, and two, Goslineria and Pharodoris, are here described. Some of these genera are more commonly found in cold or temperate waters. Austrodoris has been reported from Antarctic cold Atlantic waters (Bouchet, 1977; Wagele, 1990), Cadlina has been found in cold and temperate waters of Antarctica, South America, Japan, North America, Europe and South Africa (Rudman, 1984; Gosliner, 1987b), and Diaulula, which has only one species assigned with certainty, was de- scribed from cold or temperate waters in the Americas and Japan (McDonald, 1983). Other genera, such as Rostanga, Paradoris, Geito- doris and Baptodoris, are widespread and are not commonly found in cold regions (Baba, 1986; Rudman & Avern, 1989; Gosliner, 1991; 308 VALDÉS Miller, 1995, 1996; Ortea, 1995). Finally, only two genera, Sclerodoris and Dendrodoris, are strictly from tropical or subtropical areas (Rud- man, 1978; Valdés & Gosliner, 1999). Several hypotheses could explain the pres- ence of both tropical and cold water species in Indo-Pacific deep waters. Species of Aus- trodoris, which are absent from the tropical Indo-Pacific, are likely related to the Antarctic species. A. kerguelenensis, also found in New Caledonia Austrodoris kerguelenensis also occurs in the Antarctic deep-sea, and has a very wide geographic and bathymetric range (only in cold waters). Since the two other species found in New Caledonia share at least a portion of their geographic range with A. kerguelenensis, vicariance does not ex- plain the process of speciation that took place in this area. The species of Austrodoris de- scribed here are found in deeper and there- fore colder waters, from 500 to 1,300 m depth, whereas the rest of the species, with the ex- ception of Baptodoris phinei, were collected in shallower areas. It appears that Austrodoris is better adapted to cold waters than other gen- era and this genus occurs in large numbers in the Antarctic (Wagele, 1990) and the tropical deep sea. The tropical deep sea could be a relict area where species of Austrodoris oc- curred since the early Pliocene. Gosliner (1987a) hypothesized that during the early Pliocene, when ocean temperatures began to drop, it is likely that many species of marine organisms were widely distributed throughout the southern oceans. The oceanic warming during portions of the Pleistocene could have served as an extinction mechanism of these cold-water species in large areas of their geo- graphic ranges. The deep-water environment could have served as a refuge where Aus- trodoris species survived. On the other hand, most of the genera found in deep waters are not present in the Antarctica. In the case of Sclerodoris and Dendrodoris, the species found in New Cale- donia must have tropical sister taxa, since these genera are strictly found in tropical wa- ters. Vertical dispersal followed by subse- quent vicariance could explain the presence of these tropical species in the deep sea. Members of Sclerodoris and Dendrodoris have always been found at 460 m or less, being among the species with a shallowest occurrence. The genera Rostanga, Paradoris, Cadlina and Geitodoris have representatives in the temperate waters of New Zealand (Miller, 1980; Rudman & Avern, 1989; Miller, 1995, 1996), very close to New Caledonia, and it is possible that vicariant events were involved in the evolution of this species. Again, as Gosliner (1987a) hypothesized for South African species, the ocean warming during the Pleistocene could have served as an iso- lating mechanisms that resulted in speciation. Members of Paradoris, Cadlina and Geito- doris appear to be absent from the rest of the Indo-Pacific and also vicariance seems to be a likely explanation for the range of these species. Phylogenetic studies on each genus would determine the sister-group relation- ships and biogeographic patterns of the deep- water species here described. Most of the species studied in this paper have a pale, simple external coloration. Many of them are uniformly pale cream or white, and only a few have two contrasting colors. Bouchet (1977) described several species from Atlantic deep waters with similar color patterns. Valdés (2001) found that species of Phyllidiopsis show certain correlation be- tween their color and their collection depth. Species found deeper have simpler and paler color patterns. The coloration of shallow- water dorid nudibranchs is believed to have a strong defensive significance (Gosliner 8 Behrens, 1990). Several defensive systems based on color have been described in dorid nudibranchs, that included aposematic col- oration and mimicry, but also crypsis and spe- cial resemblance. Due to the partial or total absence of light in deep waters, the coloration of deep-sea dwellers is probably not very use- ful in an interaction with potential predators. Therefore, deep-sea nudibranchs probably rely on other defensive systems, that are also present in shallow water species, for exam- ple, chemical defense and spicules (Avila, 1995). Chemical studies have not been car- ried out on deep-sea nudibranchs, to deter- mine whether they may have higher concen- trations of toxic chemicals than shallow waters species. Since most deep-water dorids have a pale coloration, that must be some sort of ecologi- cal advantage for these animals. The most likely possibility is that deep-sea animals con- serve energy by not producing pigments that they are not going to use. Also, pale col- orations may help to camouflage species that inhabit moderately deep waters, where some light is still available. Two of the species studied in this paper lack eyes, in what also seems to be another DORID NUDIBRANCHS FROM THE DEEP PACIFIC OCEAN 309 adaptation to living in deep waters. Other blind species of nudibranchs have been pre- viously described from deep waters in the Arc- tic, Atlantic and Pacific Oceans (Wägele, 1989; Baranets, 1993; Valdés & Bouchet, 1998a, 1998b; Valdés & Bertsch, 2000). The blind species collected from New Caledonia are Austrodoris caeca and Baptodoris phinei, and both were collected at great depths, from 650 to 1,330 m in the case of the former and at 970 m in the latter. The rest of the species, which have well-developed eyes, were col- lected from areas shallower than 700 m depth. The blind Atlantic species previously described are from 1,685 and 2,538-3,016 m depth (Valdés & Bouchet, 1998a, 1998b). For this paper, 23 species have been stud- ied, of which 18 are from New Caledonia, four from the Philippines, three from Vanuatu, and one from Wallis and Futuna (the total does not add up to 23 because some species occur in more than one region). According to Philippe Bouchet (pers. comm.), New Caledonia has been sampled more extensively than any other place, and the number of records un- doubtedly reflects this sampling effort more than actual differences in absolute species richness. It is noteworthy that nearly half of the total number (11 species) have been col- lected from the Norfolk Ridge alone, a site of complex topography with extensive hard bot- toms. Only three stations, all from New Caledo- nia, had more than one species (Bathus 1 sta. CP711: Austrodoris laboutei, Pharodoris di- aphora; Bathus 3 sta. CP847: Cadlina abyssi- cola, Sclerodoris virgulata; and Halipro 1 sta. CP851: Paradoris araneosa, Dendrodoris or- bicularis). This contrasts with the experience of opisthobranch collectors in tropical shallow water, where the syntopic co-occurrence of several species of cryptobranch dorids is not exceptional. An indication that the deep-water fauna of cryptobranch dorids is still far from adequately inventoried is given by the fact that 14 species (61%) are represented by single specimens. ACKNOWLEDGEMENTS | would like to recognize the generous sup- port of several individuals. Philippe Bouchet made available the material examined in this paper, collected with the financial support of MNHN and IRD (formerly ORSTOM) during the MUSORSTOM expeditions. Specimens were collected by Bertrand Richer de Forges, Philippe Bouchet, Anders Warén and Bernard Métivier. Pierre Laboute and Jean-Louis Menou took photographs of living specimens. Katie Martin edited the final version of the manuscript. Darrell Ubick helped to prepare the samples using the critic point technique for examination on SEM. Terry Gosliner, Philippe Bouchet and Bill Rudman made con- structive comments on the manuscript. This paper has been supported by the Na- tional Science Foundation through the PEET grant DEB-9978155 “Phylogenetic systemat- ics of dorid nudibranchs” to Terrence M. Gosliner, and the Ministerio de Educacion y Cultura of Spain (SEUI), through its postdoc- toral fellowship program. Additional financial support has been made possible by the Cali- fornia Academy of Sciences. LITERATURE CITED AVILA, C., 1995, Natural products of opisthobranch molluscs: a biological review. Oceanography and Marine Biology: An Annual Review, 33: 487-559. BABA, K., 1936, Opisthobranchia of the Ryükyü (Okinawa) Islands. Journal of the Department of Agriculture, Kyúshú Imperial University, 5: 1-50, pls. 1-3. BABA, K., 1937, Opisthobranchia of Japan. Journal of the Department of Agriculture, Kyúshú Imper- ¡al University, 5: 290-344, pls. 1, 2. BABA, K., 1986, Description of a new species of nudibranchiate Mollusca, Paradoris tsurugensis, Dorididae, from Japan. Boletim de Zoologia, 10: 1-8. BARANETS, O. 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Initial attach- ment of the female bivalves occurs in April and May on small mud shrimps that have settled in mid or late winter. When the host moults, the female bivalve moves to the new exoskeleton, which allows it to grow proportionally together with the host. Shells of one year-old female bi- valves are 11.9-12.9 mm long, but few individuals attain an age of 1.5 years, corresponding to a length of 14.0-16.8 mm. By depriving the mud shrimp of part of its food, the female bivalve re- tards the growth of its host. Females = 3.2 mm are normally accompanied by several 360-um- long dwarf males attached to the bases of the host’s pereiopods. The number of males per fe- male increases during summer and autumn, but drops in December, and males are almost entirely absent during winter. Males are never transferred together with the female when the host moults, but left to perish on the old exoskeleton. Two types of sperm cells are produced. The typ- ical sperm (euspermatozoa) are stored in paired pouch-like seminal receptacles, which were previously mistaken for the species’ testes. The breeding period of females < 3/4-years old is from July through November. When breeding starts for the second time in spring, the ova be- come fertilized by sperm that have survived the winter in the receptacles. Peregrinamor ohshi- mai produces females only in spring, while males are turned out throughout spring, summer and autumn. Atypical sperm (paraspermatozoa) reside in the suprabranchial cavity outside the re- ceptacle. Their role in reproduction is uncertain. Key words: Peregrinamor ohshimai, reproduction, dwarf males, sperm dimorphism, seminal receptacle. INTRODUCTION A remarkable commensal bivalve, Peregri- namor ohshimai Shóji, has been recorded from a number of Japanese localities. It is a frequent associate of the burrowing tha- lassinidian mud shrimp Upogebia major (De Haan), more occasionally of U. narutensis (Sakai), U. yokoyai Markov, or U. issaeffi (Balss) (Miyazaki, 1936; Shdji, 1938, 1939; Sakai, 1968; Kimura & Yamamoto, 1990; Kato 8 Itani, 1995; Sakai et al., 1995; Itani & Kato, 1996). A second species, p. gastrochaenans Kato & Itani, is a commensal with U. carini- cauda (Stimpson) (Kato & Itani, 2001). The shell of р. ohshimai 15 mytiliform and may reach a length of 20 mm and a width of > 12 mm. The bivalve, always solitary, is attached to the longitudinal groove between the bases ofthe 1.-5. pereiopods of its host by means of a fan-shaped byssus. lt occupies a constant and never changing position so that its me- dian plane is parallel with that of its host and its ventral side opposed to the host. The ante- rior part of the bivalve is always directed for- ward with respect to the host (Figs. 1, 2). The systematic position of Peregrinamor has not been settled. Habe (1977) relegated the genus to the family Dreissenidae, but Kato & Itani (1995) argued that it is most closely re- lated to the Montacutidae within the super- family Galeommatoidea. They considered its mytiliform shell, otherwise atypical for this fam- ily, as an adaptation to its unique posture. Sim- ilarly to other galeommatoidean bivalves, P. ohshimai spawns its eggs into the supra- branchial chamber, where they are brooded until they become liberated as veligers. "Zoomorphology Department, Zoological Institute, Universitetsparken 15, DK-2100 Copenhagen ©, Denmark; jlutzen O zi.ku.dk. “Faculty of Education, Kumamoto University, 2-40-1, Kurokami, Kumamoto 860-8555, Japan Faculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100, Tsukide, Kumamoto 862- 8502, Japan 314 LUTZEN ET AL. 5 mm FIG. 1. Female Peregrinamor ohshimai (a) attached between the pereiopods of its host, Upogebia major, which is also parasitized by the rhizocephalan Sacculina upogebiae (b). Drawn by Beth Beyerholm. Based on section series or dissections, Shöji (1938) and Kato 4 Itani (1995) con- cluded that P. ohshimai is a hermaphrodite. If true, this would imply that this commensal, since it always occurs singly, should repro- duce by obligatory auto-fertilization, or that sufficient sperm broadcast from one bivalve into the host burrow should find its way into a specimen associated with a host in another burrow. Both alternatives seem improbable. Two paired organs claimed to be the testes are situated far back in the body. However, unlike the testes of hermaphroditic galeomma- toideans, which always constitute an inte- grated part of an ovotestis, these organs are completely separated from the ovaries in P ohshimai. We therefore decided to investigate the sexual status of Р ohshimai, resulting in the discovery that the large commensal bi- valve is a female, and that it usually lives in as- sociation with a number of hitherto overlooked dwarf males. In addition to dealing with the re- productive anatomy and sperm ultrastructure of Peregrinamor, we describe the relations be- tween the life cycles of host and commensal. MATERIAL AND METHODS The shrimps were dug up at low tide on the extensive mud flats at Kitashinchi, N. of Yat- sushiro, Kumamoto Prefecture, Kyushu. Reg- ular monthly or bimonthly samples of Upoge- bia major were taken from mid-April through mid-December 1998. Supplementary collect- ing at the same site were performed 17 March 1999 and 24 January 2000. Taking advantage of the fact that U. major is a popular bait in Japan during spring, many specimens har- bouring Peregrinamor (also originating from the Kitashinchi mud flats) were purchased live from angler’s stores 4-23 May 1999. In addi- tion, eight large alcohol-preserved, undated U. major from Matsushima hosting P. ohshi- mai were kindly placed at our disposal by Dr. Takao Yamaguchi, Aitsu Marine Biological Laboratory, University of Kumamoto. A few of the mud shrimps with bivalves were para- sitized by the rhizocephalan Sacculina up- ogebiae (Shiino) (Fig. 1). Most of the samples were preserved in 70% ethanol, but for histological and cytological purposes a representative number of bivalves were fixed in Bouin’s fluid (Sea water formula) or Karnovsky's fixative. As far as possible, the mud shrimps were sexed, and the carapace length (CL) from the tip of the rostrum to the end of the carapace determined to the near- est 0.1 mm in a petri dish on graded paper or using a calibrated eye-piece. The presence or absence of Peregrinamor females and the number of dwarf males was noted and the an- teroposterior female shell length (SL) mea- sured to the nearest 0.1 mm. Eggs spawned into the female bivalve's suprabranchial cavity could often be seen through the semitrans- parent shell. If this was not possible, the fe- males were dissected or serially sectioned. A total of 69 8-um thick, hematoxylin-eosin- stained series of paraplast sections were made of ovigerous or non-ovigerous bivalves of sizes between 3.6 and 12.9 mm SL pre- REPRODUCTION IN A COMMENSAL BIVALVE 315 FIG. 2. Female Peregrinamor ohshimai in situ on its host, Upogebia major. fb, filtering basket of host. Scale bar: 10 mm. served in Bouin's fluid or 70% ethanol. Part of a few females plus several males were em- bedded in araldite and cut into 2-um thick se- rial sections that were stained with toluidine blue. For TEM, several females and dwarf males were fixed in Karnovsky's fixative, post- fixed in a 2% osmium tetraoxide in 0.1 M ca- codylate buffer, and cut into ultrathin sections that were examined with a Jeol JEM-100X electron microscope. To study the effect of moult in Upogebia upon females and dwarf males of Peregri- nanor, individual mud shrimps with bivalves from May 1999 were placed in 200 ml beakers with running sea water and inspected twice daily throughout 14 days. Veligers released from several female Peregrinamor were cul- tured at room temperature for four weeks (7 May-4 June 1999). The larvae were kept in aerated bowls in unfiltered sea water that was changed daily. They were fed a mixture of two species of cultured algae (Tetraselmis tetrathele and Chaetoceros gracilis). The veligers and a few males were studied live and their movements video-recorded. RESULTS Population Age Structure and Growth Upogebia major: Size-frequency distribu- tion of the mud shrimps revealed the year- round presence of three cohorts. The cohort consisting of smaller specimens was always distinctly separated from the other two, which in most samples were also discrete (Fig. 3). During the studied period, the females of the two larger size-classes carried eggs from No- vember through December. No ovigerous fe- males occurred from mid April through Octo- ber. The settlement of the smallest size class in 1998 therefore occurred sometime during mid or late winter 1998, a period that is poorly represented in the samples. In April, the new cohort had grown to a size of 4-6-mm CL and by the end of December had reached 10-20 mm CL, with an average of 16 mm CL. In April the > 1-year old shrimps were 21-mm CL, and the > 2-years old ones 25-32-mm CL. The size frequency distribution of males and fe- males generally corresponded. Female Peregrinamor ohshimai: Through- out the life of the bivalve, its SL is linearly re- lated to the size (CL) of the host (Fig. 4). The smallest bivalves appeared on 15 and 27 April on 3.6-4.5-mm CL hosts, but were not mea- sured. Four 950-2,300-um SL bivalves from 12 May occurred on 3.5-6.5-mm CL U. major (Fig. 5). During summer and autumn, the bi- valves grew regularly together with the hosts and had reached an average size of 9.1-mm SL by December. When approximately 1-year old next spring (April and May), they were 11.9-12.9-mm SL. Larger bivalves were poorly represented in the samples. Among mud shrimps of the O-group, 9.56% hosted the bivalve, a figure that had decreased to one-fifth, or 1.88%, among animals of the 1- group. According to information from people used to collecting preferentially large Upoge- 316 LUTZEN ETAL. 2 GROUP i] ot LGROUP 7 > males Upogebia major CL (mm) \ \ ---- females АРК MAY JUNE JULY AUG SEP ОСТ NOV DEC FIG. 3. Growth curves for Upogebia major calcu- lated from average CL of bimonthly samples April through December 1998. Kitashinchi, Amakusa ls- land. 207 mm) 107 Peregrinamor ohshimai SL Upogebia major CL (mm) FIG. 4. Relationship between sizes of Peregrinamor ohshimai and Upogebia major, April through De- cember 1998 and May 1999. The line is repre- sented as SL = -0.954 + 0.680CL, r2 = 0.824 (P < 0.0001). Kitashinchi, Amakusa Island. bia for bait or food, large bivalves are very rare. Fifteen 14.2-15.6 mm large bivalves from September through November were probably approximately 1.5-years old. Judg- ing from the size of the host (29.2-mm CL) and their own length (17.0 and 18.3-mm SL), two specimens of Peregrinamorfrom July and August must have been more than 2 years old. Eight more large undated shells (16.6- 20.0-m SL) associated with very large hosts (28.0-34.7-mm CL) were probably the same age. Male and female mud shrimps were associ- ated with the commensal bivalves to the same degree (7.8%, respectively 7.7%). For each of the months April through December, the aver- age CL of Upogebia of the 0-group hosting P. ohshimaiwas constantly 1-2 mm shorter than that of the total material of hosts (Fig. 6). The Males When the female bivalves were cautiously lifted from their contact with the host or re- moved entirely, a number of tiny male bivalves were usually seen to be attached by byssus threads to the basis and coxa of the host's pereiopods or to the narrow sternal groove in between. For attachment, the fourth and fifth pereiopods facing the posterior part of the fe- male's mantle slit were preferred to the three foremost pairs. Occasionally, a few males were located on the female's shell margin or on its byssus. The shape and size of the shell of the dwarf males vary within narrow limits (Figs. 14, 15). It is regularly oval in shape, very slightly in- equilateral, semitransparent, approximately 360 ит long and 305 um high and laterally compressed. The hinge has two subsymmet- rical lateral teeth. The foot is relatively large and partially cili- ated. The pedal glands are prominent (Figs. 7, 8). Conspicuous anterior and posterior byssus retractor muscles are associated with the byssus gland. If live males are removed from the shrimp, they move around quickly. They do this by extending the foot to more than the shell length, stick its tip to the sub- stratum, then drag the body along. Males were occasionally seen to detach voluntarily and move from place to place on and between the limb bases. The gills are represented by inner demibranchs. A short, ciliated oesopha- gus terminates within a mass of undifferenti- ated yolk-rich endodermal cells. No other parts of the alimentary tract were demonstra- ble. All major ganglia are present, and a pair of statocysts lie adjacent to the large pedal ganglia. A prominent testis occupies the entire visceral mass and has paired openings pos- teriorly in the mantle cavity. Even if of constant SL, the males vary con- siderably in the development of the testes and some other organs. The specimen illustrated in Figure 7 represents an immature stage with a large 150 x 200 um testes demonstrating the early stages of spermiogenesis. The mass of yolk-rich cells is very extensive and the ad- ductor muscles are well developed. Figure 8 shows the terminal stage in the life of the males. The testis has shrunk to a fraction of its original volume and is now filled with mature REPRODUCTION IN A COMMENSAL BIVALVE 317 1 GROUP Average no. males per female > 1 GROUP Я 14 2 Sid E 1 Sogn: Br 112 D a 2 > un оО Е Ro) 8 Gp = 6 3 > 4 < 1 GROUP SOTBUI YJIM зэ[еита} % 0 GROUP зэтеиэу SNOLASIAO Y, JAN FEB MAR APR MAY JUNE JULY AUG SEP OCT NOV DEC FIG. 5. Peregrinamor ohshimai. Frequency of males throughout the year and growth and reproduction of fe- males. Figures along the growth curve (c) indicate sample sizes (female p. ohshimai). All data from Kitash- inchi, Amakusa Island, except for those marked by an asterisk from Kato & Itani (1995) and Miyazaki (1936). Upogebia major CL (mm) = — without Peregrinamor ohshimai with Peregrinamor ohshimai APR MAY JUNE JULY AUG SEP OCT NOV DEC FIG. 6. Average size of Upogebia major with (n = 283) and without Peregrinamor ohshimai (n = 3,071), April through December 1998 (0-group). Ki- tashinchi, Amakusa Island. spermatozoans. The posteriorly directed, short spermiduct becomes clearly visible in live specimens. Characteristically at this Stage, both adductor muscles have de- creased in volume, and the mass of endoder- mal cells has degenerated into a very small body. These two types, including any interme- diary stages, may occur on the same mud shrimp hosting a female bivalve. The number of males estimated to be sexually mature was small, however, and in ten mud shrimps ex- amined hardly exceeded 10%. Up to 53 males were recorded with a single female bivalve, but their number was usually much smaller (Fig. 9). The frequency of males was expressed in two ways: (1) as the per- centage of female bivalves associated with males, and (2) as the average number of males per female (Fig. 5). Apart from a very few males recorded in late June, they first be- came associated with the O-group of females in late July. The smallest female co-occurring with males was 3.2 mm. Males became in- creasingly abundant in August when they oc- curred together with most of the females, al- though the number per female was still moderate. Their frequency peaked during au- tumn, when a high number (15-20) of males had associated with almost every female bi- valve. The samples from December showed a distinct drop in their number. Only one male 318 LUTZEN ET AL. 100 mm FIGS. 7-8. Peregrinamor ohshimai. Anatomy of an immature (7) and sexually mature (8) dwarf male, left side view. aa, anterior adductor; ab, anterior byssal retractor; b, byssus; bg byssus gland; cg, cerebral gan- glion; f, foot; h, heart; id, inner demibranch; k, kidney; |, ligament; mm, mantle margin; oe, oesophagus; pa, posterior adductor; pb, posterior byssal retractor; pg, pedal ganglion; sm, shell margin; st, statocyst; t, testes; ud, undifferentiated tissue; vg visceral ganglion. Drawn by Beth Beyerholm. 2mm FIG. 9. A female Peregrinamor ohshimai (left side view) with its six dwarf males. is, extended inhalent siphon. Drawn by Beth Beyerholm. was found together with seven females of the 1-group from January through April. The males reappeared in early May, where on an average 9.7 males were associated with all of 23 females. In general males were few or absent on large Upogebia associated with large female bivalves. Thus, males were only recorded to- gether with two among eight 16.6-20.2 mm long female P. ohshimai. The collecting date of these specimens were not known. The Females Whether small, medium-sized or large, the female's shell is always attached singly with its ventral shell margin facing the host’s sternal groove and in such a way that its anterior shell margin is level with the basis of the first pair of pereiopods (chelipeds, Fig. 2). This is also true with the smallest shells (950-2,300 um SL) that were placed far anteriorly on the ventral part of the thorax between the first pair, or the first and second pairs of pereiopods. The shell of the smallest female (SL 950 um) is inequilateral, rounded, shortened be- hind, and with a distinct approximately 350- um-long prodissoconch (Fig. 10). There are two subumbonal laterals. The surface is pol- ished and marked by week growth lines. Ina 1,500 um long specimen (Figs. 11, 12), the shell has started to become laterally inflated, the posterior part has elongated to exceed the front part in length, and the ventral margin has become almost straight. In older shells, growth chiefly takes place in the posterior part of the shell, which in a 2,300-um-long speci- men (Fig. 13) had grown to double the length of the front part. When 3-4 mm long, the bi- valves had acquired the mytiliform shape, typ- ical of the species. A distinct ovary occupies a major portion of the anterior mantle and a small part of the foot, exactly as described by Shóji (1938) and Kato & Itani (1995). At each side, the ovary is divided into a large lateral portion and a much smaller elongate dorsal posteriorly directed portion. The two parts fuse just behind the middle of the body in a short and narrow lobe, which terminates medially at the wall of the suprabranchial cavity. In the absence of dis- tinct oviducts, this is presumably where a tem- REPRODUCTION IN A COMMENSAL BIVALVE 319 400 um FIGS. 10-15. Peregrinamor ohshimai. FIG. 10, left shell of 950 tum long female. FIGS. 11, 12, left shell and hinge of right shell of 1,500 um long female. FIG. 13, left shell of 2,300 um long female. FIG. 14, inside of right shell of male. FIG. 15, live specimen of male in left view. p, prodissoconch; t, testes; ud, undifferentiated tissue. Drawn by Beth Beyerholm. porary opening appears when the eggs are spawned into the incubatory cavity. Histologi- cally, the ovary consists of many branching tubular lobules. Besides a stock of oogonia along the wall of the lobule, the ovary contains two more or less distinct size classes of oocytes, which undergo vitellogenesis simul- taneously. When ripe, paraplast-embedded sectioned oocytes have a diameter of 85-90 um, corresponding to 100-105 um measured in newly spawned live eggs. In females pre- served immediately before emission of the larvae, the next, and larger, generation of ovarial oocytes have almost attained their maximum size, and the even smaller genera- tion of oocytes vary between 25 and 45 um in diameter. Since at the peak of the reproduc- tive season (August-October) only three out of four sexually mature bivalves were oviger- ous (Fig. 5), there must be an interval sepa- rating the emission of the larvae and the next spawning, in which the larger oocytes grow to their final size. The lay-out of the reproductive organs was studied in many specimens from 3.5 to 12.9 mm. Irrespective of the size of the specimens, a testis, whether part or not of the ovary, was absent. It is obvious, however, that the small, paired organs believed to be the testes by previous authors, are receptacles for the stor- age of sperm (Figs. 16, 17). Lying superficially far posteriorly within the mantle, they were visible as two usually whitish curved bodies through the thin, transparent shell. Each formed an oblong sinuous sac that is located close to the respective visceral ganglion. It had a narrow ciliated opening into the suprabranchial chamber (incubatory cham- ber) far posterior to the oviducts. The opening was placed within the margin of a small, raised area of the suprabranchial chamber wall lined with a tall, cylindrical, ciliated ep- ithelium (Fig. 16). The wall of the seminal receptacle also consisted of a cylindrical, cili- ated epithelium bearing numerous 2-3-um- long slender microvilli (Fig. 20). The seminal receptacles were typically filled with fully de- veloped flagellated euspermatozoa orien- tated with part of their long, thin heads em- bedded in the surface of the cells of the lining epithelium (Figs. 20, 21). In all sizes exam- ined, there were absolutely no signs of sper- matogenesis. In addition, masses of sperm of another type, paraspermatozoa, with a much shorter head were usually present in the su- prabranchial cavity close to, but outside the seminal receptacles. We conclude that the large specimens of P. ohshimai are females and that at oviposition into the incubatory chamber the ova become fertilized by sperm cells discharged from the receptacles. While sperm were normally present in the 320 LUTZEN ET AL. FIGS. 16-21. Peregrinamor ohshimai. Histological sections through the seminal receptacles filled with eu- spermatozoa (16 and 17); heads of paraspermatozoa (18) and euspermatozoa (19); Figs 20, 21, electron- micrograph showing aligned euspermatozoa embedded in the epithelial cells of the receptacle. 2 um thin sections of araldite embedded material stained with toluidine blue (16) and 8 um thick paraplast sections stained with H + E (17-19). a, acrosome of euspermatozoon; cr, ciliary root; d, duct of seminal receptacle which opens into the suprabranchial cavity (su) near arrow; e, embryo; ep, ciliated epithelial cell of recepta- cle with microvilli (m); n, nucleus of euspermatozoon; o, ovary; om outer mantle epithelium; p, parasperma- tozoa, rc, raised ciliated area for reception of paraspermatozoa; sr, seminal receptacle. Scales represent 50 um (16), 100 um (17), 10 um (18 and 19), 2 um (20), and 1 um (21). REPRODUCTION IN А COMMENSAL BIVALVE 321 receptacles of females that had passed 3.6 mm SL, the oocytes did not become mature until at a SL of 5.5-6.0 mm. With the excep- tion of two 5.5-mm large ovigerous speci- mens, female Р ohshimai usually did not spawn until having attained a SL of 6.0 mm. Between a SL of 6.0 mm and 6.9 mm oviger- ous and non-ovigerous females were equally numerous, but when > 7.0 mm, there were ap- proximately three times as many ovigerous as non-ovigerous specimens. The reproductive activity, expressed as the percentage of ovigerous females, excluding specimens < 6.0 mm, started in the latter half of July with a maximum in August, September and October. Reproduction declined again in November and had come to a complete stop in early De- cember, when none of the 36 females col- lected were ovigerous. Many females of the 1- group had spawned in early May, but the percentage was not recorded. Through the semitransparent female shell, it could be observed that during incubation in the suprabranchial cavity, ova and embryos were rotating vigorously, probably moved by the action of the ciliated gill epithelium. Some of the ovigerous females that were kept alive in the laboratory liberated straight-hinged lar- vae 7 May. The shells of freshly released lar- vae measured 205 um in length and 152 um in height. One week-old larva had grown to 250 um and 200 um in height and length, but did not increase further in size. At that stage the larvae mostly rested on the bottom and only now and then lifted from the substratum. They were still alive, but immobile, when at an age of 25 days the incubation was discontin- ued. The Sperm Peregrinamor ohshimai produces two types of flagellated sperm, euspermatozoa and paraspermatozoa (terminology of Healy 4 Jamieson (1981)). These types correspond broadly to the categories “typical” or “eu- pyrene” spermatozoa (= euspermatozoa) and “atypical” or “oligopyrene” spermatozoa (= paraspermatozoa) often used in older litera- ture. The euspermatozoon has a slender, al- most straight 9.5-10.0-um-long head, whereas the head of the paraspermatozoon is shorter, 4.5 um long, but broader and slightly conical (Figs. 18, 19). Both types were found in squash preparations of live males. Within the female’s receptacles, only euspermato- zoa were found (Figs. 16, 17). The parasper- matozoa were either attached with their heads to the raised, ciliated epithelium of the suprabranchial cavity near the receptacle’s orifice (Fig. 17), or occurred as masses of free cells in the adjacent part of the suprabranchial cavity, perhaps because they had been de- tached as a response to fixation. When unat- tached, the paraspermatozoa were often mixed with smaller amounts of euspermato- zoa. Spermatids of paraspermatozoa often occurred together with mature sperm. Euspermatozoa were present in all of 69 sectioned females from 3.6 to 12.9 mm SL, except for a single 3.8-mm-long specimen. In two specimens (3.6 and 4.4 mm SL), they oc- curred in only one of the two receptacles. They were abundantly present in all females from winter (December through April). Paraspermatozoa were absent in most (N = 13) of the smaller bivalves < 5.9 mm SL (N = 16) and in all but one of seven females from January, March and April. In many sectioned bivalves with SL from 6.0 to 11.4 mm collected from July through December various quanti- ties of paraspermatozoa occurred in approxi- mately 75% of the specimens. Behaviour of Females and Males at Moulting of the Host Kato & Itani (1995) demonstrated that dur- ing the moulting process of its host, Peregri- namor is able to detach itself from the old ex- oskeleton and move to the new exoskeleton and reattach in the proper location. We con- firmed this behaviour in several cases. To study the fate of the dwarf males, which were unknown to Kato & Itani, we compared their number on the old and new exoskeletons of seven isolated recently moulted hosts. Of a total of 44 dwarf males, only one accompa- nied the female to the new exoskeleton, and this was probably because it had originally at- tached to the female’s byssus. In three other cases, where the presence of males before moulting, but not their number, were noted, again none of them were transferred to the new exuvia. Simulating a moult, in six other mud shrimps the female bivalves (SL 11.6-12.1 mm) were removed by cutting the byssus, and the associated males were in- spected immediately following the operation and 12 hours later. Their reaction to the fe- male’s removal was slight, as 38 of a total of 45 males present remained in their original position, three had moved to beyond the coxae of the pereiopods and maxillipeds, and 322 LUTZEN ETAL. only four had left the host. In addition, we oc- casionally noted that mud shrimps with bi- valves and with an unusually clean exoskele- ton, consistent with a recent moult, were always devoid of any dwarf males. DISCUSSION Upogebia major lives in a U-shaped burrow excavated in the mud flats. Its four pairs of ab- dominal swimmerets generate a steady cur- rent of water through the burrow. The water current is being filtered by a fine-meshed bas- ket composed by numerous long close-set setae of its maxillipeds and first two pairs of pereiopods and maxillipeds (MacGinitie, 1930; Mukai & Koike, 1984). Kato & Itani (1995) studied the feeding of the female Р ohshimai. When the shells gape, the right and left anterior mantle edges are extended to form an inhalent siphon that penetrates into the host's filtering basket (Fig. 9). The species is thus a true commensal, which feeds upon suspended matter retained in the host’s bas- ket. The constantly smaller average CL of Up- ogebia hosting P. ohshimai (Fig. 6) seems to show that the bivalve removes a substantial part of the host’s food. To reach into the filter- ing basket and not at the same time obstruct the movements of the pereiopods, or become damaged or even crushed by them, the exact position along the host’s midline cannot be compromised (Fig. 2). This is probably why two or more bivalves were never seen to co- inhabit the same host individual in the several hundreds of instances of this association recorded in the literature or seen by us. Many galeommatoidean bivalves are protandric hermaphrodites and in most of the cases where dwarf males have been re- ported, they are merely considered as an initial stage in such a life cycle and capable later of transforming into a functional female (O Foighil, 1985a). All evidence suggests that P. ohshimai is strictly gonochoristic. The males that have attained sexual maturity are completely exhausted and evidently have no potential to develop further. Also no traces of any testes were ever found in the smallest fe- males examined histologically. Gonochorism in Р ohshimai is probably a secondary adap- tation to the species’ extremely sedentary and solitary habits. The males are so small that, even when plentiful, they do not interfere with the female’s position. Neither do they com- pete for food, since, living in a water current that has been filtered by the host's food bas- ket and having no functional alimentary tract, they must depend wholly on the energy re- sources accumulated in their own tissues. It must be assumed that the males are fairly short-lived, but that dead or lost individuals are being replaced by a supply of new ar- rivals. Morphologically the dwarf males of P ohshimai show a striking resemblance to the dwarf males (complemental males) of Monta- cuta phascolionis (Dautzenberg & Fischer) discovered by Deroux (1961a). The female’s particular feeding posture also means that small specimens, as was al- ways the case, have to be placed with their anterior shell gape close to the base of the fil- tering basket to allow the short inhalent siphon to reach into it. The reason for the ob- served linear correlation between sizes of the female bivalve and the host (Fig. 4), also ob- served by Kato & Itani (1995), suggests that efficient food uptake by the bivalve requires a fairly constant size-relation between the in- halent siphon and the food basket. The clear separation of the size-classes of the host allows a fairly accurate determination of its age from the size. Because P. ohshimai only settles upon small hosts and grow pro- portionately together with them, there is also no overlap between the size-classes of the commensal (Fig. 5). Relatively many more Upogebia of the O-group were hosting Pere- grinamor than those of the 1- and 2-groups. Kato & Itani (1995) found 29.2% of Upogebia of the O-group from September to harbour the bivalve, compared to 18.4% among speci- mens of the 1-group in April. Combined with the small number of records of large bivalves, this seems to show that the number of the commensals is greatly reduced before or while entering its second year, which is well in advance of the time when the host becomes sexually mature (November-December). This is a short life span compared to other com- mensal bivalves, which may attain a maxi- mum age of from four to seven years (Franz, 1973; Ockelmann & Muus, 1978). There are at least two separate reproduc- tive periods during the life of Р ohshimal. Specimens of the 0-group commence spawn- ing in July, when they pass the critical 6.0 mm SL, but the almost constant presence of eu- spermatozoa in the receptacles in specimens > 3.6 mm SL is evidence that insemination oc- curs much earlier. Breeding continues through November but comes to a complete stop in December (Fig. 5). Of 41 females from REPRODUCTION IN A COMMENSAL BIVALVE 323 December through March, only one specimen (from January) was ovigerous, but all sec- tioned ones had very large oocytes in the ovary and the receptacles full of sperm. Fe- males of the 1-group started to reproduce again in April, when all of nine 8.5-16.7 mm SL specimens were “producing eggs and brooding developing eggs and veligers” (Kato & Itani, 1995). The appearance of rather newly settled female Peregrinamor in April and May, and their absence in June, is also consistent with this conclusion. Many of the 1- year old bivalves were ovigerous in early May, and so were three of four specimens from early June (Miyazaki, 1936) but there is no more information from the summer months. The freshly released larvae (205 x 152 um) are slightly smaller than reported by Miyazaki (1936, as Erycina sp.), namely 220 x 170 um. Miyazaki succeeded in incubating the larvae to the pediveliger stage, in which the shell measured 351 x 298 um. This corresponds to the size of the prodissoconch of the female shell as well as to the shell size of the dwarf males (360 x 305 um). Apparently, larvae de- veloping into both males and females settle upon the mud shrimps immediately after they have completed the planktonic phase. While there is a good correlation between the presence and abundance of dwarf males and the reproduction in the females of the 0- group, some males are still present, although at a reduced number, when reproduction comes to a stop in early winter (Fig. 5). The reproductive role of these males is probably largely played out and they cannot be com- pensated for as the production of larvae has ceased. In keeping with this, only a single male was found associated with the seven fe- males from January, March, and April. When spawning is resumed in spring, fertilization of the ova evidently depends upon the sperm that has been stored for three to four winter months within the receptacles. From the lar- vae produced by the 1-group in the spring are recruited (1) the next 0-дгоир of females, (2) males that become associated with the 0- group in June-July, and (3) the first males of the 1-year old female bivalves, noted first in early May. As no settling of female bivalves could be demonstrated during the breeding period of the O-group, it follows that its sole contribution to the population is to produce males. This is apparently also the role of those females of the 1-group that survive long enough to spawn during summer and autumn. In conclusion, P. ohshimai produces females only in spring, while males are turned out throughout spring, summer and autumn. The very high number of males compared to females partly compensates for their small size and individually limited sperm production. Upogebia major undergoes a number of moults throughout summer and autumn, and at each moult any males present are lost. Since males nevertheless increase in number during this period, lost males not only become replaced, but additional males are being added. The number of moults in U. major is not known, but in the smaller Mediterranean U. pusilla (Petagna) there are 7-9 moults per year (Dworschak, 1988). Assuming the num- ber of moults in U. major to be of the same order and the observed annual average of males per female of Р ohshimai at 10-15 (Fig. 5), the number of males produced for each female is probably several times higher. Seminal receptacles in bivalves are known only in Xylophaga dorsalis (Turton) (Pho- ladoidea: Pholadidae) (Purchon, 1941) and in the family Montacutidae. All montacutids are brooders that incubate the ova in the suprabrancial cavity, with which the recepta- cles are associated. Among the Montacutidae there are three types of receptacles: In species of Jousseaumiella they are claimed to arise as pouches from the oviduct (Bourne, 1906; Knudsen, 1944). Nipponomysella sub- truncata (Yokoyama) is peculiar in the pos- session of two mushroom-shaped recepta- cles, which protrude from the visceral mass into the incubatory cavity (Lútzen et al., in press). In Litigiella glabra (Fischer), M. phas- colionis, Mysella cuneata (Verrill & Bush), and Potidoma subtrigonum (Jeffreys), the recep- tacles are paired pouches from the floor of the suprabranchial cavity (Pelseneer, 1911; Der- oux, 1961b; Gage, 1968; Jespersen & Lutzen, 2000). Very similar pouches, so-called acces- sory male organs, are found in Mysella biden- tata (Montagu) (Deroux, 1961b; Ockelmann & Muus, 1978). The sperm depositories in PR. ohshimai obviously belong to the same type. Since the species is strictly gonochoristic, it is certain that the sperm stored in the receptacle and in the adjacent part of the suprabranchial cavity are foreign sperm. Similarly to M. bidentata, which have sperm depositaries as well as dimorphic sperm, only euspermatozoa were found within the receptacles of P. ohshi- mai. lt remains to be settled whether the sperm are transferred from the males in sperm bags, such as may occur in some other montacutids (Deroux, 1961b; Ockelmann & 324 LUTZEN ET AL. Muus, 1978; O Foighil, 1985a,b; Lútzen et al., in press). Stored sperm enable P. ohshimai to repro- duce uninterruptedly even when males are absent. When the host moults, which proba- bly happens several times during the bivalve’s breeding season, the female bivalve becomes dissociated from the accompanying males. Some time will elapse until new males settle and become sexually productive, but in the in- terval the stored sperm allow the female to re- produce at its convenience. During the winter months, males are totally absent, and re- sumption of the reproduction in spring in the 1-group is dependent on the euspermatozoa that have survived for 3-4 months in the re- ceptacles. Paraspermatozoa are obviously less viable, as they occurred only sparsely during winter. There are no other data to show how long sperm of bivalves may remain func- tional. But in some prosobranch gastropods, sperm may still be capable of fertilization for more than one year (Coe, 1942; Chi & Wag- ner, 1957). Compared to the condition in Proso- branchia, sperm dimorphism is rare in bivalves and confined to two species of the family Mon- tacutidae (Mysella bidentata; Montacuta te- nella Loven) and the Thyasiridae (Axinodon symmetros (Jeffreys)) (Ockelmann, 1965). The list now includes also P. ohshimai. The significance of paraspermatozoa in molluscs is enigmatic. Ockelmann & Muus (1978) sug- gested that they perform some signal func- tion. Because they seem to be absent when the females of P. ohshimai resume spawn- ing in spring, they probably take no part in fertilization. Popham (1940) stated that the Montacuti- dae are characterized by lack of the outer demibranch. A vestige of the outer demi- branch is present in Peregrinamor (Shóji, 1938), but it is also in the montacutid Monta- cutona (Morton, 1980). The discovery that Peregrinamor has seminal receptacles and dimorphic sperm, until now only known from species of the Montacutidae, lends support to the view of Kato & Itani (1995) that the genus is most closely related to this family. ACKNOWLEDGEMENTS We are very grateful to Dr. Ase Jespersen, Department of Zoomorphology, Copenhagen, for her assistance in operating the electron microscope. We would also like to thank Dr. Keiji Baba, Faculty of Education, Kumamoto University, and Dr. Takao Yamaguchi, Aitsu Marine Biological Station, Kumamoto Univer- sity, for providing us with specimens of P ohshimai. 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SHOJI, K., 1939, On the curious commensal bi- valve (Peregrinamor ohshimai Shóji) which at- taches to Upogebia major. Japanese Journal of Zoology, 51(2): 101 (In Japanese). Revised ms. accepted January 3, 2001 MALACOLOGIA, 2001, 43(1-2): 327-336 EYEBALLS AND PITFALLS: ESTIMATING THE DEGREE OF HETEROSTROPHY IN THE HETEROBRANCH SHELL (GASTROPODA: HETEROBRANCHIA) Roberto Cipriani’ ? & Rüdiger Bieler' ABSTRACT Heterostrophic gastropod shells, in which the coiling axes of proto- and teleoconchs diverge, pose interesting ontogenetic questions and complex morphological characters for systematic analyses. The inclination angle of the protoconch in relation to the axis of coiling of the teleo- conch has been used to distinguish between otherwise similar heterostrophic species. The pre- cision of this metric, however, is very sensitive to measurement biases. This paper introduces a new technique, the box method, which provides a more precise estimate of the angle of inclina- tion of the protoconch. A rectangular box is drawn around an image of the protoconch; the box diagonal then is used as an estimator of the angle of inclination. The accuracy of the technique is investigated with a simulation model that calculates two-dimensional projections of realistic three-dimensional coiling morphologies. The results of the model suggest that the studied met- ric is dependent on the morphology of the protoconch and on its degree of immersion in the first teleoconch whorl. Caution is in order when basing taxonomic decisions on differences in the in- clination of the protoconch without considering the underlying morphologies and without know- ing the error associated with these differences. Given that the accuracy of this metric cannot be achieved or known, the precision of this angular measure should be maximized and its limits demonstrated. Authors are encouraged to include in their reports a description of the technique used to estimate the protoconch inclination and to give; a measure of the error of the angular measurement made for each particular taxon or sample.” Key words: Gastropoda, lower Heterobranchia, shell morphology, shell coiling, protoconch, angle measurement, statistics, precision. INTRODUCTION A key feature and putative synapomorphy of the Heterobranchia — a large clade of Gas- tropoda encompassing the traditional groups of Pulmonata, Opisthobranchia and various former “prosobranchs” such as Pyramidel- loidea, Architectonicoidea, and Valvatoidea — is the phenomenon of heterostrophy (Hasz- prunar, 1985, 1988; Bieler, 1992; Ponder 4 Lindberg, 1997). In other groups, the proto- conch (embryonic and larval shell) and teleo- conch (postlarval shell) are coiled around the same axis, resulting in a homeostrophic shell. Heterostrophic shells, by contrast, display a divergence between the coiling axis of proto- conch and teleoconch (Fig. 1). [For helpful reviews of the terminology associated with coiling variability in gastropods, including het- erostrophy, see Cox (1960: 110-120) and Robertson (1993).] The obvious cases of heterostrophy are those in which a dramatic change of coiling occurs just before (e.g., Bandel, 1995; Cipri- ani, 1997) or at metamorphosis (e.g., Robert- son, 1985, 1993), resulting in noticeable di- vergence of the coiling direction/axes between protoconchs and teleoconchs. Het- erostrophy is particularly well expressed in basal heterobranchs —termed “lower hetero- branchs” in recent works—such as Pyra- midellidae, Architectonicidae, and Mathil- didae. Certain authors (e.g., Golikov & Starobogatov, 1975; Ponder & Warén, 1988; Bieler, 1992) have used Fischer’s (1885) ex- pressive name “Heterostropha” for this proba- bly paraphyletic group of taxa. More recently, the name Heterostropha has also been used synonymously with Heterobranchia (e.g., Bandel, 1990; Schróder, 1995; Grúndel, 1998). In these strongly developed cases of het- ‘Department of Zoology, Field Museum of Natural History, 1400 S. Lake Shore Drive, Chicago, Illinois 60605-2496, U.S.A.; and Committee on Evolutionary Biology, University of Chicago; bieler@fmnh.org “Current address: Departamento de Estudios Ambientales, Universidad Simon Bolivar, Caracas, Venezuela; send mail to: CCS 91392, P.O. Box 025323, Miami, Florida 33102-5323 U.S.A.; rcipri@usb.ve This paper is dedicated to Dr. Robert Robertson of The Academy of Natural Sciences of Philadelphia, in recognition of his pioneering work on the morphology of heterostrophic shells. 328 CIPRIANI 8 BIELER Y? KEIL TIEREN EN FIG. 1. Protoconch and early teleoconch shell of same specimen of Mathilda cf. amanda (New Caledonia; protoconch diameter of 460 um), viewed from two different angles. erostrophy, the protoconch is sinistrally coiled, but the larva is anatomically dextrally organized. The apparent sinistrality is a result of hyperstrophy (i.e, the inverted “upward” coiling along the axis, as opposed to coiling “downward” in regular orthostrophy). Other lower heterobranchs, such as Cornirostridae, Valvatidae, Orbitestellidae, Omalogyridae and Hyalogyrinidae, show only initial hyper- strophy (Bieler et al., 1998), a weak expres- sion of heterostrophy that affects only the ini- tial whorl(s) of the larval shell. An_ initial left-coiling of the first part of the protoconch whorl is also reported for the vetigastropod families Trochidae (Hadfield & Strathmann, 1990) and the extinct Discohelicidae (Schröder, 1995). Heterobranch protoconchs, in contrast to the situation in Caenogastropoda, display few morphological features in shell pattern and sculpture. Among extant taxa, such features are usually limited to sutural wrinkles, aper- tural varices, anal keels and associated cal- luses (e.g., Robertson, 1964; Bieler, 1993), with more elaborate patterns restricted to members of a few groups, such as the pyra- midellid genus Chrysallida (e.g., Penas & Rolan, 1998). In the absence of ornamental characters, differences in coiling parameters thus provide valuable morphological charac- ters used in heterobranch systematics. The traditional metric of heterostrophy is the incli- nation angle of the protoconch body whorl in relation to some fixed morphological refer- ences, usually the axis of coiling of the teleo- conch (Micali, 1992; Bieler, 1995; Schróder, 1995). It is important to note that some au- thors give the smallest angle between the two hypothetical shell axes, rather than the de- gree of “turn”; a 25° divergence reported by Bandel (1995), for instance, corresponds to 155° of other authors. An extreme case of heterostrophy exists in which the degree of heterostrophy is so great that the protoconch assumes an upside-down position on the teleoconch —in widely umbili- cated forms actually protruding from the um- bilicus, rather than the spire, of the teleo- conch. This condition, for which Dautzenberg & Fischer (1896) introduced “anastrophie” and Minichev & Starobogatov (1971) “coxial heterostrophy,” is prevalent in the family Ar- chitectonicidae and a few other groups. Char- acters of heterostrophy have thus been used at various taxonomic levels, from inferring membership to the Heterobranchia based on heterostrophic coiling (e.g., Bandel, 1994) to assigning membership to a particular family, genus, or species. As a systematic character at a lower taxo- nomic level, the inclination angle has been used to distinguish between otherwise similar species. As long as the recognized ranges of average measures of variability (e.g., vari- ance) are significantly different between the compared taxa, this character is useful for comparison and phylogenetic analysis. For in- stance, the angle appears to be near-constant within extant Indo-Pacific species of Mathilda (Bieler, 1995) and, in congruence with other morphological characters, often differs among species. By contrast, Lower Cretaceous species of the same genus were described with intraspecific variability ranging up to 35° (Schróder, 1995). Wise (1996) employed the protoconch angle as a character in his phylo- genetic analysis of Pyramidellidae, using nar- rowly delimited character states of 90-95”, ESTIMATING THE DEGREE OF HETEROSTROPHY 329 A Diagonal B Average Line Lines running parallel to walls of body whorl of protoconch FIG. 2. Box method. This method measures variables using a rectangular framework enclosing the proto- conch. Protoconch is rotated around coiling axis of teleoconch to estimate minimum inclination of box diag- onal from axis of teleoconch coiling. Use of this framework reduces measurement error, increasing precision of inclination angle measurement. Variables used in this method include (A) angle of diagonal of box, and (B) average angle of two lines running parallel to walls of body whorl of protoconch. 120-125°, 130-135°, and 140-145°. Schan- der et al. (1999) used wider ranges for their character states of 90-100° and 100-145° in a subsequent analysis. Most modern taxo- nomic studies involving heterostrophic shells now provide detailed descriptions of the incli- nation angle, the number of protoconch whorls, and most recently also of the degree of immersion into the first teleoconch whorl (e.g., Hori, 1998). Several practical questions arise when using heterostrophy as a character in hetero- branch systematics: (1) How accurate/precise are such measure- ments made with the traditional method? How great is the measurement error? (2) Can the traditional method be improved, thus reducing the measurement error? (3) Is the degree of heterostrophy, as ex- pressed by “degree of inclination of the protoconch against the inferred axis of the teleoconch,” a useful/meaningful charac- ter when comparing morphologies in which the protoconch has different de- grees of overlap by the first teleoconch whorl? The present paper attempts to answer these questions. MATERIALS AND METHODS To address our objectives, we used a het- erostrophic mathildid species, identified as Mathilda cf. amanda Thiele, 1925 (Hetero- branchia: Mathildidae), for which such mor- phological parameters as the angle of inclina- tion of the protoconch are easy to measure. Representatives of this morphological variety have previously been described from the Indo-Pacific (Bieler, 1995); the exemplar for this study was collected in the Eastern Atlantic (Sénégal, Region de Dakar; Marche-Marchad Sta. 9, 20 February 1957; Muséum National d’Histore Naturelle, Paris [MNHN]; not num- bered). The protoconch of this species is rela- tively large (about 500 um in maximum diam- eter), has approximately 1.5 whorls, and is only half covered by the first teleoconch whorl. The 12 whorls of the teleoconch mea- sure 20.5 mm in height. This species was cho- sen to give a minimum estimate of measure- ment error, and measurements of species with less exposed protoconchs will likely be less accurate. All measurements were taken from the same shell. This implies that all the errors discussed are introduced by the re- searcher, by the methodology, or by its use on the shell, and are not due to differences be- tween or within species. Also, all measure- ments for this study were taken by a single ex- perimenter (RC); any human error is assumed consistent throughout repeated measure- ments. To compare the results of the traditional technique (Fig. 2) with those produced by the other techniques that follow, the angle of incli- nation of the protoconch was always esti- mated between the body whorl of the proto- conch and an imaginary line running parallel to the upper suture line of the first teleoconch whorl. To report this angle in traditional fash- 330 CIPRIANI 8 BIELER Angle of inclination of protoconch | B Angle of inclination of first teleoconch whorl Axis FIG. 3. Using box method, estimating maximum angle of protoconch from axis of coiling is equivalent to mea- suring minimum angle between diagonal of box and lower border of box. Protoconch is rotated around teleo- conch axis. In each step of rotation, inclination of protoconch is measured, and corresponding value is used as sample’s angle. After all protoconch angles from same sample are collected at different instances of ro- tation, value of minimum angle is selected as most repeatable measure. Rotation sequence starts from po- sition A. Protoconch displays smallest angle in position B. ion, the angle between this imaginary line and the axis of coiling of the teleoconch was mea- sured and added later to the first angle (see example below). Given that all measurements were taken from the same individual, it can safely be assumed that the error of this cor- recting angle is constant and therefore is not affecting the relative magnitudes of the mea- surement errors from the different protocols described below. The traditional technique was emulated by measuring the inclination of the protoconch from an orientation that ap- peared to show maximum angle to the eye. In an attempt to improve the estimation ac- curacy for the inclination angle of the proto- conch, a simple alternative we call the box method was developed and tested. This ap- proach uses a framework of reference (i.e., a rectangular box) around a two-dimensional image of the protoconch to overcome the problem of the absence of morphological landmarks on the shell. The lower border of this rectangle is aligned with the upper suture of the first teleoconch whorl. The upper border of the rectangle is a line parallel to the lower border, and separated from it by a perpendic- ular distance large enough to contain the pro- toconch body whorl between both lines. The lateral sides close the rectangle to contain the protoconch (Fig. 2). Two variables inside this box were used as guidelines to measure the inclination angle of the protoconch: (a) the di- agonal of the rectangle aligned with the body whorl of the protoconch, and (b) the average angle between two lines that run parallel to each side of the walls of the protoconch body whorl (Fig. 2). To further reduce the measurement error, the specimen was first mounted on a support with a rotating axis. This axis was mounted perpendicular to the plane of sight of a dis- secting microscope. The shell was attached to the tip of the rotating axis using a small quantity of modeling clay at the base of its body whorl. The coiling axis of the shell was aligned to the axis of rotation by correcting the shell's relative orientation until the off-center oscillations of its spire were minimal. The shell was positioned with its apex pointing toward the viewer. Then, the shell was rotated until the axis of coiling of the protoconch was ap- proximately at 45° of inclination from the plane of sight. The protoconch and first two teleoconch whorls were outlined using a drawing tube attached to a dissecting micro- scope, with a magnification large enough to produce drawings of the protoconch with a di- ameter of approximately 4 cm on the paper. In all drawings, the centerline of the teleoconch (i.e., axis of coiling) was approximated by es- timating the midline of the two lines extending through the lateral extremes of the teleoconch sutures. An outline was drawn again after the shell was rotated 15° in the direction that made the axis of the protoconch more colin- ear with the plane of sight (Fig. 3). This pro- ESTIMATING THE DEGREE OF HETEROSTROPHY 331 cedure was repeated at least four or five times, or until the shell rotated approximately 180° around its own axis. In each outline, the heterostrophic angle was measured between this line and the lower border of the box. The shell was removed from the support and mounted again, and the procedure repeated each time a complete sequence of angular measures was taken. For comparative purposes, we also mea- sured the angle of inclination of the proto- conch of a single outline, using the diagonal line of the box framework described above, once the shell was rotated to a position that seemed to display the greatest inclination to the experimenter’s eyes. This angle was cal- culated by using the following trigonometric formula: diagonal where lateral and diagonal are the lengths of the lateral side and the diagonal of the box, re- spectively. Averages were estimated using two to ten replicates. A rarefaction analysis, with 500 bootstraps per replicate, was used to estimate the magnitude of measurement error ex- pected to occur for a given number of repli- cates. Acomputer program was developed to help us understand how the morphology of the pro- toconch and its immersion in the first teleo- conch whorl affects the accuracy of the box method. This program simulated two-dimen- sional projections of a protoconch on top of a teleoconch in a large number of morphologi- cal combinations, given the coiling parame- ters that matched those found in this and other samples of heterobranch gastropods (Cipriani, unpubl.; program available at http://prof.usb.ve/rcipri). The program repre- sented the cross section of the body whorl of a “rotating” protoconch with respect to the teleoconch axis, simulating different angles of inclination, distances between tube cross- sections, tube diameters, immersion of the protoconch into the first teleoconch whorl, and inclination of the first teleoconch whorl in rela- tion to its coiling plane. The accuracy of the angular estimation (i.e., the diagonal of the box frame) in relation to the “real” angle of the protoconch was compared with the per- centage overlap of the protoconch by the first teleoconch whorl and the inclination of the lat- ter in relation to its coiling plane. The “real” lateral Angle = arcsin angle of inclination of the protoconch was de- fined as the angle between the line passing through the two centroids of the tube cross- sections and the axis of teleoconch coiling. The amount of overlap was defined as per- centage of the diameter of the largest cross section of the protoconch body whorl (with simulated percentages of 0%, 35%, 70%). The inclination of the first teleoconch whorl was simulated at 0°, 20° and 40° from the plane of coiling of the teleoconch. It is important to note that what is actually simulated in this model is a measuring tech- nique on a two-dimensional projection of a shell. This is exactly what practicing systema- tists do when aligning the shell under the mi- croscope to measure the angle of inclination of the protoconch. Hence, the results of this model should be insightful on the potential bi- ases of the measuring technique. RESULTS AND DISCUSSION The maximum absolute deviation of the angle from one measurement taken of the specimen of Mathilda cf. amanda, estimated using the traditional method, was 10.5° (or +5.3°) around its average (X = 32.7°). The av- erage deviation of this measurement, ex- pressed as a standard error, was +2.5° (Fig. 4). These numbers mean that after two angu- lar measurements are taken from this individ- ual of Mathilda cf. amanda, the actual estima- tion can be on average any value between 30.2° and 35.2° (Fig. 4). Considering the max- imum deviation of this measure, the estima- tion could actually be 27.4° to 38.0°. The rare- faction analysis suggests that it would be necessary to estimate an average of five an- gular measurements of the same sample to reduce the variability by 50%. If the shell is rotated, and the minimum angle of the box diagonal is taken from each set of rotations, the precision of the angular measure is readily increased. Using this method, the value of a single measurement is expected to be on average +0.7° of the aver- age angular value (Fig. 4). The maximum de- viation in this experiment ranged between +1.4° around its average. This means that at least eight replicates would be necessary to reduce the error of the traditional technique to a value similar to that obtained from averag- ing two replicates using the box method. 332 CIPRIANI & BIELER Angle Replicates FIG. 4. Estimations of angle of inclination (in degrees) of body whorl of protoconch of Mathilda cf. amanda in relation to line defined by lower border of box frame (i.e., line defined by upper border of first teleoconch whorl). Results of traditional technique (№) and box method (6). Standard error margins estimated from rar- efaction analysis (500 replicates) identified by finely dashed (box method) and continuous lines (traditional technique). If instead of the diagonal variable the aver- age angle between both lines around the body whorl is measured, the standard error increases to +1.4° (Fig. 5) and the maximum deviation to +3.1°. This variability is similar to that obtained if the diagonal of the box is used but the shell is not rotated (standard error = +1.7°; maximum deviation = +3.3°) (Fig. 5). Calculating the diagonal of the box using a trigonometric approach resulted in angles with errors identical to those obtained directly measuring the inclination of the diagonal line (standard error = +0.7°; maximum deviation = +1.4°). These errors are not too large consid- ering the error reported in the traditional tech- nique, even though the position of the shell is subjectively chosen to measure the inclination of the protoconch in these protocols. How- ever, of all of the above, those angles mea- sured using the minimum diagonal value on a rotating shell were the most precise. Even if the average inclination of the protoconch body whorl seems a more natural approxima- tion to the angle of inclination than the diago- nal of the box, the precision of the former is strongly dependent on the morphology of the body whorl and of the relative immersion of the protoconch. Different approaches were used to reduce this variability, including se- lecting positional landmarks to help trace the lateral lines, but none of them were success- ful. The accuracy of the angle is harder to measure. At a first glance, the average of the estimated angles varies with the method (Figs. 4, 5). This implies that each methodol- ogy has its own biases, and these have to be part of the criteria taken into account by the researcher when choosing a particular proto- col. The results of the computer simulation suggest that the morphology of the proto- conch biases the estimation of its angle of in- clination using the box method. The accurate estimation of the angle using the diagonal of the box is strongly dependent on the percent- age of protoconch overlap and on the inclina- tion of the first protoconch whorl in relation to its plane of coiling. Notice, in Figure 6, that the estimated inclination does not match the ac- tual angle (i.e., straight line), and is strongly dependent on morphological characters such as the protoconch overlap. These results imply that: (a) different angu- lar measurements may come from proto- conchs that actually are identically inclined, and (b) even using the same technique, two measured angular values that are equal may actually come from protoconchs that are dif- ferently inclined. Consequently, caution is suggested when basing taxonomic decisions on differences in the inclination of the proto- conch without considering the underlying morphologies and knowing the error associ- ESTIMATING THE DEGREE OF HETEROSTROPHY 333 Angle 1 2 3 4 5 6 i 8 9 10 11 Replicates FIG. 5. Estimations of angle of inclination (in degrees) of protoconch body whorl of Mathilda cf. amanda in relation to line defined by lower border of box frame. Plot shows angle values of diagonal line (@) and of av- erage inclination of two lines fitted on sides of the protoconch (№), both obtained from box method applied without rotating shell. Rotational position of shell from which measurements are taken is approximated by eye. Standard error margins estimated from rarefaction analysis (500 replicates) identified by finely dashed (diagonal) and continuous lines (average). ated with these differences. Measuring angles from drawing-tube sketches introduces errors in the estimation, but direct measurement of the inclination of the protoconch from the mi- croscope ocular limits the use of a reference frame, thereby increasing the variability of the metric. One potential way to reduce the error caused by the drawing-tube approach is using a video or digital camera system connected to the microscope to load the shell image (or scanning micrograph) into a computer system before measuring the angle. However, this approach will not eliminate or reduce angular errors associated with the measuring method itself. Moreover, different technologies used to reproduce the images introduce their own errors; scanning electron micrographs, for in- stance, can be notoriously distorted. In this particular case, accuracy and preci- sion seem to have an inverse relationship; we, however, do not possess objective infor- mation about the accuracy of techniques other than the box method. Despite of all the problems outlined above, the metric traditionally used to describe het- erostrophic shells can still be of use for local taxonomic purposes, as long as its error is taken into account. Given that the accuracy of this metric cannot be improved, the precision of this angular measure should be maximized and its limits understood. In particular, it would be very useful to indicate to other researchers not only a description of the morphology of the protoconch and first teleoconch whorl, but also a description of the technique used to es- timate the inclination of the protoconch, and overall, the precision of the measurement made of each particular taxon or sample. These values should include averages and the errors of such averages, or at the very least the range of values taken from the sam- ple. How should this error be reported? If the angle estimation takes place between the morphological features of interest (e.g., proto- conch body whorl and axis of coiling), then the error could be expressed using different parameters, such as variance, standard devi- ation, or intervals of confidence. However, if the sum of two or more separate angles is re- quired, the measures of error from those av- erages have to be treated accordingly. The following is the rationale of this approach: The errors associated with the function F (x, y) of the sums of the averages (e.g., angles of coiling axis and protoconch with lower border of box), are part of the variability of the (as- sumed) normal distribution to which the actual function F (x, y) is asymptotically tending (when n > +). Given that HONEY 334 250 275 Predicted Angle 250 275 250 275 300 300 300 CIPRIANI & BIELER 325 350 375 325 350 375 325 350 375 Angle of Protoconch FIG. 6. Results of computer simulations (N = 900). Inclination of protoconch of Mathilda cf. amanda esti- mated using box method, as a function of protoconch overlap by first teleoconch whorl: (A) 0%, (B) 35%, and (C) 70%. Values of inclination of first teleoconch whorl expressed in degrees: = 0; @ = 20; A = 40. Per- centages expressed on basis of diameter of largest protoconch cross section. Black diagonal lines represent values in which predicted angle and real angle match, for each of the overlapping percentages. Notice that accuracy of box method (i.e., intersection of simulated values and straight lines) varies depending on per- centage of overlap and inclination of first teleoconch whorl. where x is the average angle of the proto- conch with the upper suture line, and y is that of the suture line with the axis of coiling. Given that x and y are independent, then following Ku (1966), 0? [Е (x, У] = 01 + oY where о? is the parameter estimator of the population variance, IN 2 1 AS n=1 and nis the number of samples. For example, the average minimum angle of the first ten measurements of the sample's protoconch obtained from the box method was 30.4” and its variance 1.1”. Recall that this is the angle between the protoconch body whorl and the base of the box, and that the base of the box is inclined with respect to the plane perpendicular to the axis of coiling of the teleoconch. Estimation of the angle be- tween the base of the box and this plane re- sulted in an average of 9.9”, with a variance of 3.4” (n=10). Hence, X = 90 - (30.4 + 9.9) = 49.7”, [x] = 1.1 + 3.4 = 4.5°. Only the variances from the averages are summed (1.1 + 3.4), because 90° is added with no error (by definition in this model). This constant represents the angular difference between the axis of coiling of the teleoconch and the plane perpendicular to it. The stan- dard deviation of the average is olx] = V о"[Х= 2.1° and the 95% confidence interval, using the critical values of Student's t-distribution for n= 10 (Rohlf & Sokal, 1981), is 2.262 - o[x] Vn The result accordingly is reported as x + o[x] 0.95 [9] — = 1.5°. ESTIMATING THE DEGREE OF HETEROSTROPHY 335 Omens 2915) ога X = Los (49.17 = 1.57). Given the small sample size used in this ex- ample, confidence intervals could also be es- timated using a non-parametric method. The dependence of the inclination of the protoconch on other morphological shell vari- ables makes it practically impossible to obtain accurate results, or even worse, to know if the estimations are accurate at all. However, the concept of accuracy is strongly dependent on the qualities of the variable used to measure the desired morphology. An important ques- tion can be raised about the adequacy of this metric to describe such morphology. This question is beyond the scope of this paper, al- though one must be aware that the morphol- ogy of heterostrophic shells is relatively com- plex. Given this morphological complexity and the difficulty of assigning landmarks to the whorls, it is ambitious to think that heterostro- phy can be summarized in a single parameter. However, these results are not intended to discourage systematists from using the tradi- tional metrics of heterostrophy. The intention of this paper is to expose some of the limits of this metric, and to promote a better under- standing of its statistical properties and con- sequent use. ACKNOWLEDGMENTS The current paper resulted from discussions and experiments of the authors to address practical problems in comparative morpholog- ical studies of heterostrophic gastropod shells. The focus on heterostrophic coiling led to in- depth morphological, physical, and evolution- ary studies as part of a Ph.D. dissertation by RC, the results of which will be published else- where. RC wishes to thank Mike LaBarbera, Gene Hunt, Mike Foote, David Jablonski, and Susan Kidwell for their useful comments and criticism. Philippe Bouchet (MNHN) made this and many other specimens of lower hetero- branchs available to our studies. Two anony- mous reviewers provided valuable comments on an earlier draft of the manuscript. Lower heterobranch gastropod research was sup- ported by Hinds Fund awards from the Com- mittee on Evolutionary Biology and grants from Sigma Xi and the Society of Western Malacologists to RC, as well as NSF DEB- 9318231 to RB, and NSF DEB-9509324 to RB and Timothy Collins (Florida International Uni- versity). LITERATURE CITED BANDEL, K., 1990, Shell structure of the Gas- tropoda excluding Archaeogastropoda. Pp. 117- 134, in In: J. G. CARTER, ed., Skeletal biomineral- ization: patterns, processes and evolutionary trends, Vol. 1. New York, Van Nostrand Reinhold. BANDEL, K., 1994, Triassic Euthyneura (Gas- tropoda) from St. Cassian Formation (Italian Alps) with a discussion on the evolution of the Het- erostropha. Freiberger Forschungsheft, C452: 79-99, incl. pls. 1-4. BANDEL, K., 1995, Mathildoidea (Gastropoda, Het- erostropha) from the Late Triassic St. Cassian Formation. 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Journal of Research of the Na- tional Bureau of Standards — С. Engineering and Instrumentation, 70C(4): 263-273. MICALI, P., 1992, Due nuovi Pyramidellidae (Mol- lusca: Gastropoda) Pliocenici: Chrysallida varisculpta n. sp. e Folinella spinosula n. sp. Bol- lettino Malacologico, 28(5-12): 195-202. MINICHEV, YU. S. & YA. |. STAROBOGATOV, 1971, Heterostrophy and its significance in the evolution of Gastropoda. Pp. 10-12 in: |. M. LIKHAREV, ed., Molluscs: Trends, Methods and some Results of their Investigation. Moscow, Academy of Sciences of the U.S.S.R., Zoology Institute [in Russian]. PENAS, A. 4 E. ROLAN, 1998, La familia Pyra- midellidae Gray, 1840 (Mollusca, Gastropoda, Heterostropha) en Africa Occidental. 3. El género Chrysallida s. |. Revista de la Sociedad Española de Malacología, Iberus, Suppl. 4: 1-73. PONDER, W. Е. & D. В. LINDBERG, 1997, Towards a phylogeny of gastropod mollusks: an analysis using morphological characters. 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Ophelia, 51(1): 39-76. SCHRODER, M., 1995, Fruhontogenetische Scha- len jurassischer und unterkretazischer Gastropo- den aus Norddeutschland und Polen. Palaeonto- graphica, (A)238(1-4): 1-95, pls. 1-15. WISE, J. B., 1996, Morphology and phylogenetic relationships of certain pyramidellid taxa (Hetero- branchia). Malacologia, 37(2): 443-511. Revised ms accepted 2 January 2001 MALACOLOGIA, 2001, 43(1-2): 337-347 TAXONOMIC REVISION OF PISIDIUM PUNCTATUM STERKI, 1895 (BIVALVIA: SPHAERIIDAE) Alexei V. Korniushin', Igor A. Grigorovich? & Gerald L. Mackie? ABSTRACT The status of Pisidium punctatum Sterki, 1895, is revised based on the examination of type material and new collections from the lower Great Lakes. Heterogeneity of the shell and anatom- ical characteristics suggests that this taxon is a composite of two different species. The two va- rieties of P. punctatum described from the Great Lakes basin — P. punctatum $. str. and P. punc- tatum var. armatum Sterki, 1905 —are synonymized with the Eurasian native P. moitessierianum Paladilhe, 1866, because they all possess crescent-shaped folds around the umbones, similar hinge morphology, poorly developed radial mantle muscles, and a profoundly reduced dorsal lobe of the nephridium. Pisidium punctatum var. simplex Sterki, 1905, previously established as a form of Р punctatum, is redescribed and raised to species rank. Pisidium simplex is distin- guished from P. moitessierianum by the absence of umbonal folds and by having a more elon- gated shell, less extensively developed hinge teeth, and more pronounced mantle muscle bun- dles. In addition, P. simplex appears to have a much broader distribution in North America than P. moitessierianum. Pisidium simplex resembles the Central American native P. punctiferum (Guppy, 1867) in mantle musculature and nephridium shape, suggesting a close affinity between these taxa. Lectotypes of P punctatum s. str., P. punctatum var. armatum, and P. punctatum var. simplex are designated. Key words: bivalve mussel, Pisidium, Sphaeriidae, Great Lakes, cryptogenic species. INTRODUCTION The North American fauna of small fresh- water clams of the family Sphaeriidae is char- acterized by considerable taxonomic diversity —36 species belonging to four genera —and close affinities to the European fauna (Her- rington, 1962; Burch, 1975). Nine sphaeriid species — Musculium lacustre (Müller, 1774); Sphaerium nitidum Clessin, in Westerlund, 1876; Pisidium casertanum (Poli, 1791); PR conventus Clessin, 1877; P. lilljeborgi Clessin, 1886; P. milium Held, 1836; P. nitidum Jenyns, 1832; Р obtusale (Lamarck, 1818); and P. subtruncatum Malm, 1855 —have broad Hol- arctic distributions. Four Eurasian species found in North America, S. corneum (Lin- neaeus, 1758), Р amnicum (Müller, 1774), P. henslowanum (Sheppard, 1823), and P supinum Schmidt, 1851, have most records from the Great Lakes-St. Lawrence basin, in- dicating their nonindigenous origin in New World (Herrington, 1962; Clarke, 1981). One North American species, Musculium transver- sum (Say, 1829), was apparently introduced to England (Ellis, 1978). Most of the work done on invading freshwater invertebrates in North America has focused on the bivalves Dreissena polymorpha, D. bugensis, and Cor- bicula fluminea (D'ltri, 1997; Nalepa & Schloesser, 1993). Much less effort has been directed toward the study of the Sphaeriidae, the invasions of which may involve a small number of colonists. The affinity of the North American Pisidium punctatum s.|. to European species has been repeatedly suggested. Herrington (1954) treated it as a senior synonym of P. tenuili- neatum Stelfox, 1918, whereas Ellis (1978) and Piechocki (1989) tentatively syn- onymized it with Р moitessierianum Pal- adihle, 1866. On the other hand, Herrington (1962) suggested that P. punctatum is a syn- onym of Central American P. punctiferum (Guppy, 1867). Kuiper (1962b) and later re- viewers (Burch, 1975; Clarke, 1981) recog- nized P. punctatum as a valid species. Descriptions of P. punctatum provided by "Institute of Zoology, National Academy of Sciences of Ukraine, Kiev, 01601, Ukraine Corresponding author. Great Lakes Institute for Environmental Research, University of Windsor, Windsor, Ontario N9B 3P4, Canada; gigoruw @ uwindsor.ca “Department of Zoology, University of Guelph, Guelph, Ontario NIG 2W1, Canada 338 KORNIUSHIN, GRIGOROVICH 8 MACKIE different authors are rather controversial and cannot help to resolve the problem of its identity. Sterki (1895) pointed out that the species possesses concentric umbonal folds, whereas Kuiper (1962b) noted that these folds were similar to those in P. moitessieri- anum. However, a distinct variety of P. punc- tatum lacking folds was described under the name simplex Sterki, 1905. Clarke (1981) ob- served the folds in many specimens and con- sidered them a characteristic feature of P. punctatum, whereas Mackie et al. (1980) did not include this character in their description of the species. Anatomical characters used for discrimination of European species of Sphaeriidae (Korniushin, 1996, 1998) were not yet applied to the North American taxa. Considering these discrepancies in the litera- ture and lack of soft tissue characters, a re- view of the systematics of this species is clearly needed. In this study, we examine the Sterki’s type material and recent collections of P. punctatum $.|. from the lower Great Lakes and compare the species’ conchological and anatomical characteristics with those of similar species from Europe and Central America. The mor- phological heterogeneity of P punctatums.|. is demonstrated. Pisidium punctatum s. str. and P. punctatum var. armatum Sterki, 1905, are synonymized with Р moitessierianum Pal- adihle, 1866, whereas P. punctatum var. sim- plex Sterki, 1905, is raised to a full species. The status of Р moitessierianum in North America and affinities to P. simplex are briefly discussed. MATERIAL AND METHODS Our revision was initiated in the course of surveys in the St. Clair River-western Lake Erie corridor conducted during 1997-1998. Both folded and non-folded forms of P. punc- tatum were collected, and their soft anatomies were studied alongside the traditional shell characters. Syntypes series of P. punctatum s. str, Р punctatum var. simplex, and P. punc- tatum var. armatum were revised, along with other relevant collections of Sterki. Lecto- types were selected from the mentioned type lots in order to ensure consistent use of these names in the future. In all cases, lectotypes retained catalogue numbers of the original syntypes series (provided with species’ de- scriptions); the new paralectotypes’ numbers are also provided, when applicable. The lots of P. moitessierianum collected earlier in vari- ous basins of Ukraine and reviewed by Kor- niushin (1996) were used for comparison. Ma- terials of Р punctiferum were obtained from the Field Museum of Natural History, Chicago, and from the private collection of Claus Meier- Brook, Tubingen, Germany. Voucher speci- mens were deposited in the American Mu- seum of Natural History, New York, New York, and the Field Museum of Natural History, Chicago, Illinois. The following parameters were measured on a stereoscopic optical microscope using a calibrated ocular micrometer: shell length, shell height, shell width, height of hinge plate, length of hinge (distance between the cusps of the lateral teeth in the left valve), length of ligament, and distance between the umbo and the posterior shell edge. Because some morphological features of adults may differ in young Pisidium, all measurements were taken from the largest adults with a shell length > 1.3 mm. Several indices were calculated, including the elongation index (height to length ratio), the convexity index (width to height ratio), and the hinge index (hinge height to shell height ratio). Descriptive statistics (mean values and standard deviation) and 2-tailed t-tests were executed using standard statistical software. Scanning electron microscopy (SEM) was conducted on a JEOL microscope in the Mu- seum fúr Naturkunde, Berlin, Germany. Hinge teeth nomenclature corresponds to that in Herrington (1962). Specimens for anatomical study were fixed in 70% alcohol or in 5% formalin and dis- sected under a stereomicroscope. Nephridia were observed in situ, gills and mantle prepa- rations were stained by Grenacher’s carmine and water eosine, respectively, and mounted in Canada balsam for microscopic study (for details, see Korniushin, 1996). Abbreviations: aa, anterior adductor; as, anal siphon; bp, brood pouch; c, ctenidium; C2, inner cardinal tooth of left valve; C3, car- dinal tooth of right valve; C4, outer cardinal tooth of left valve; dg, digestive gland; dl, dor- sal lobe; f, foot; H, shell height; hh, hinge height; hl, hinge length; L, shell length; Il, lat- eral loop; Is, ligament size; Ip, labial palps; lup, distance from the umbone to the posterior shell end; mre, outer radial mantle muscles; mri, inner radial mantle muscles; n, nephrid- ium; pa, posterior adductor; pd, pericardial tube; sr1, retractor of anal siphon; sr2, retrac- tor of branchial siphon; W, shell width. TAXONOMIC REVISION OF PISIDIUM PUNCTATUM Museum acronyms: CMNH, Carnegie Mu- seum of Natural History, Pittsburg, Penn- sylvania, U.S.A.; FMNH, Field Museum of Natural History, Chicago, Illinois, U.S.A.; MCZ, Museum of Comparative Zoology, Cambridge, Massachusetts, U.S.A.; MHNG, Museum d'Histoire Naturelle de Geneve, Switzerland. SYSTEMATICS Pisidium moitessierianum Paladilhe, 1866 Figs. 1-4 Pisidium moitessierianum Paladilhe, 1866: 172; Kennard & Woodward, 1926: 323 (European synonymy); Favre, 1943: 43, fig. 23 Pisidium punctatum Sterki, 1895:99, pl. 2 figs. 7-12; Kuiper, 1962b: 173-178, figs. 7-9; Ellis, 1978: 82 (probable synonymy with P moitessierianum) Pisidium punctatum var. armatum Sterki, 1905: 84 Pisidium punctiferum (Guppy): 1962: 47-48 (part) Herrington, Type Material Pisidium punctatum Sterki, 1895 - two syn- types MCZ 112563, Tuscarawas River, Bear Run, tributary of Mahoning River, Portage Co., Ohio, lectotype here designated (Fig. IAS 15, hl = 14, W = 1.0 [measure- ments in mm], paralectotype’s new accession number MCZ 319616. Pisidium punctatum var. armatum Sterki, 1905 - >15 syntypes CMNH 4642, Tus- carawas River, Navarre, Stark Co., Ohio, Coll. V. Sterki, 28 October 1903, lectotype here designated (Fig. 1B) - L=1.6, H=1.4, W= 1.0 [measurements in mm]. Other Materials Examined CMNH 11692, Tinkers Creek, Bedfort, Cuyahoga Co., Ohio, Coll. H. Wright [det. as Pisidium punctatum var. simplex,] July 1927. CMNH 6688, Ohio River below Portsmouth, Ohio, Coll. V. Sterki, September 1910. West- ern Lake Erie, Michigan, Coll. I. A. Grig- orovich, 27 May 1998. St. Clair River delta, Ontario, Coll. I. A. Grigorovich, 10 August 1997. Lake St. Clair, Ontario, Coll. |. A. Grig- orovich, 4 June 1998. Dnieper River near Kiev, Ukraine, Coll. A. V. Korniushin, July | oO ==> FIG. 1. Shells of Pisidium punctatum s. str. and P. punctatum var. armatum. A, P. punctatum s. str. Lectotype, MCZ 112563 (external frontal and lateral and internal lateral view of left valve and hinge fea- tures of single individual). Bear Run, Ohio; B, P. punctatum var. armatum. Lectotype, CMNH 4642 (external frontal and lateral and internal lateral view of left valve and hinge features of single individual). Tuscarawas River, Ohio. Scale bar = 1 mm. 1987. Lake Beloye, Volyn Region, Ukraine, Coll. A. V. Korniushin, August 1998. Description Shells (of North American specimens) small (maximal L = 1.9 mm), triangular or trapezoid, relatively short and high (elongation index 0.86-0.92), moderately or markedly convex (convexity index 0.7 to 0.8), with flat, not protruding subcentrally placed umbones and characteristic crescent umbonal folds of same form as in European P. moitessierianum (Figs. 1A, B, 2, 3). Surface sculpture with fine, but somewhat irregularly spaced concen- tric ribs. Scars of mantle muscles not pro- nounced, hardly distinguishable from mantle line (Fig. 1). Hinge plate relatively broad, arched. Cardi- nal tooth in right valve (C3) bent at its middle point and bifurcated posteriorly (in European specimens arched). Inner cardinal tooth in left valve (C2) of triangular shape; outer cardinal tooth slightly bent. Lateral teeth strong, some- what swollen. Ligament pit enclosed (not visi- ble from the dorsal side), relatively broad. 340 KORNIUSHIN, GRIGOROVICH 8 MACKIE Internal pores noticeable in many, but not all North American specimens; pores not found in European specimens (Figs. 2C, D, 3C). Gills, or ctenidia (Fig. 4A, B). Outer demi- branch absent. Brood pouches (one per gill) placed dorsally and formed by nine filaments. Five to ten larvae per gill. Mantle edge (Fig. 4C). Inhalant siphon ab- sent. Exhalant siphon with one pair of retrac- tors. Retractors of reduced inhalant siphon also noticeable. Inner radial muscles of pedal slit grouped in five noticeable but rather weak bundles. Outer radial mantle muscles rela- tively long. Nephridia (Fig. 4D) small; dorsal lobe evi- dently reduced, low and broad, only slightly cleft; lateral loop visible from outside. Distribution Type locality: Maurin, near Montpellier, France (Paladilhe, 1866). This species is re- ported from many European countries includ- ing Great Britain (Ellis, 1978), Germany (Glóer & Meier-Brook, 1998), Poland (Piechocki, 1989), Ukraine, Belarus and Russia (Kor- niushin, 1996). In North America, it is known from Lake Erie and St. Lawrence drainage (Sterki, 1895), as well as St. Clair River and Lake St. Clair (Grigorovich et al., 2000). Remarks In erecting P. moitessierianum from Maurin specimens, Paladihle (1866) indicated the fol- lowing diagnostic characters: crescent folds around umbones, well-developed concentric surface ribs on shell, and a strong hinge. Since Kennard & Woodward (1926), the name Pisidium moitessierianum has been adopted in European literature (Ellis, 1978; Piechocki, 1989; Gloer & Meier-Brook, 1998). Favre (1943) redescribed and figured a spec- imen from Maurin (MHNG, Bourguignat coll. #375) presumed to be one of syntypes. Sphaeriids from the Great Lakes and Ukrain- ian drainages correspond to the descriptions of P moitessierianum by Paladihle (1866) and Favre (1943). The combination of concholog- ical features, principally the existence of well- developed umbonal folds, strong shell sculp- ture and hinge characters, points conclusively to the two varieties of P. punctatum, namely punctatum s. str.and armatum, as being iden- tical to Р moitessierianum. Values of the shell indices of the typical (folded) form of P. punctatum also agree with those of the European Р moitessierianum (Table 1). Possessing a relatively narrow hinge plate, especially under the umbone, North American specimens of Р moitessieri- anum are most similar to specimens from Lake Beloye in Ukraine. Individuals from the European rivers (e.g., Dnieper River near Kiev; Table 1, Fig. 3) are characterized by a somewhat stronger hinge and arched cardinal teeth (Ellis, 1978; Piechocki, 1989). Anatomi- cal characters of the Great Lake specimens do not differ from those of European P. moitessierianum as described by Korniushin (1996). Pisidium moitessierianum is similar in many shell and hinge characters to P. cruciatum, but umbonal folds in the latter species are arched, with ventrally diverging branches. Compared to young specimens of P. compressum and P. lilljeborgi cristatum, which may also have crescent folds, P. moitessierianum differs in its smaller size and stronger hinge. Pisidium simplex Sterki, 1905 Figs. 5-7 Pisidium punctatum var. simplex Sterki, 1905: 84 Pisidium punctatum: Kuiper, 1962b: 173-178, fig. 6 Pisidium punctiferum (Guppy): Herrington, 1962: 47-48 (part) Type Material Syntypes (>15 specimens) CMNH 2804, Lillycash Creek, Joliet, Illinois, Coll. J. H. Handwerk, 1899, lectotype here designated (Fig. 5A) - L= 1.8, H = 1.5, W = 1.2 [mea- surements in mm]; syntypes (one complete specimen and two separate valves) CMNH 272, Fox River, Wisconsin, Coll. G. T. Marston, No. 202, P.102;. Other Materials Examined CMNH 10121, Lake Washington, Seattle, Washington, Coll. C. C. Engberg, 1922. CMNH 10563, Beaver Lake, Washington, Coll. C. C. Engberg, 1924. CMNH 10047, Whatcom Lake, Bellingham, Washington, Coll. C. C. Engberg, 1922. CMNH 3658-b, Crystal Lake, Benzie Co., Michigan, Coll. R. J. Kirckland, 1902. CMNH 3781, Brook near In- TAXONOMIC REVISION OF P/SIDIUM PUNCTATUM 341 FIG. 2. Shell and hinge morphology of the Great Lakes populations of Pisidium moitessierianum. Lake St. Clair, Ontario: A, shell. Arrow indicates an umbonal fold; B, external shell sculpture; C, hinge; D, western Lake Erie, Michigan, hinge and internal shell surface with pores. Scale bars = 0.1 mm. dian mounds, Kent Co., Michigan, Coll. R. J. Kirckland, 1902. CMNH 8224 Locust Creek, Bolo, Illinois, Coll. A. A. Hinkley, 3 September 1908. CMNH 7107, Hudson, Ohio, Coll. V. Sterki, May 1909. CMNH 5933, Beacoup Creek, Du Bois, Illinois, Coll. A. A. Hinkley, 1907. CMNH 7856, Wabash River, Chairs, In- diana, Coll. A. A. Hinkley, 1908. CMNH 6207, Tinker’s Creek, ditch, 2 miles east of Hudson, Ohio, Coll. V. Sterki, 27 May 1909. CMNH 3273, Tuscarawas River, Ohio, Coll. V. Sterki, September 1897. CMNH 3278, Erie Canal, Hamilton, Ohio, Coll. V. Sterki, August 1901. CMNH 6198 Tinker’s Creek, 3 km east of Hudson, Ohio, Coll. V. Sterki, 27 May 1909. CMNH 6213, Brandy Wine Creek, Hudson, Ohio, Coll. V. Sterki, May 1909. CMNH 12867, Lake Erie beach, Cedar Point, Sandusky, Ohio, Coll. V. Sterki, July 1919. CMNH 269, brook, Rootstown, Ohio. CMNH 6515, Lake Erie, Presque Island, Pond Lake, Pennsylva- nia, Coll. V. Sterki, July 1910. CMNH 6462, Lake Erie, ditch to pond on Big Bend, Presque Island, Pennsylvania, Coll. V. Sterki, July 1910. CMNH 271, Herkimer Co., New York, Coll. A. Bailey, 1894. CMNH 8357, Oneida Lake, South Bay, New York, Coll. F. C. Baker, 1916. CMNH 7557 North Warren, Maryland, Coll. N. W. Lermond, August 1913. Western Lake Erie, Michigan, Coll. I. A. Grigorovich, 27 May 1998. CMNH 10399, Lake Ontario near Toronto, Ontario, Coll. D. B. Adamstone, 1922. CMNH 10351, Lake Nipigon, Nipigon channel, Ontario, Coll. D. B. Adamstone, 1922. St. Clair River delta, Ontario, Coll. |. A. Grigorovich, 4 June 1998. Description Shell somewhat larger than in North Amer- ican Р moitessierianum (maximal L = 2.0 mm), oval, more or less elongated (elongation 342 KORNIUSHIN, GRIGOROVICH 8 MACKIE FIG. 3. Shell and hinge morphology of the Euro- pean population Pisidium moitessierianum. Dnieper River, Ukraine. A, shell. Arrow indicates an umbonal fold; B, external shell sculpture; C, hinge. Scale bars = 0.1 mm. index less than 0.85), rather convex (convex- ity index up to 0.86), with relatively broad, pro- truded, posteriorly placed umbones (Figs. 5, 6). No umbonal folds. Upper margin arched, other margins evenly curved. Surface sculp- ture variable: from very delicate to rather pro- nounced, regular striations. Scars of mantle muscles clearly distinguishable, elevated over mantle line (Fig. 5A). Hinge plate very thin, arched. C2 slightly bent, C3 and C4 almost straight, C3 cleft on posterior end. Lateral teeth delicate, not swollen. Ligament pit narrow; ligament of closed specimens somewhat elevated over upper margin of hinge plate. Pores present. Gill and brood pouches (Fig. 7A, B) simi- lar to those in the former species. Mantle (Fig. 7C). Siphons as in previous species. Inner radial mantle muscles form seven well-developed bundles. Outer radial muscles relatively short. Nephridia with well-developed, deeply cleft dorsal lobe; lateral loop visible from outside (Figs. 7D). Distribution Type locality: Lillycash Creek, Joliet, Illinois. This species is broadly distributed in North America, from the Atlantic to the Pacific coast. Remarks This species differs from P. moitessierianum in that it lacks the umbonal folds and pos- sesses an elongated shell with markedly asymmetrical umbones, a narrow hinge plate with almost straight cardinal teeth, more nu- merous and strongly developed mantle mus- cle bundles and nephridia with a rather high and deeply cleft dorsal lobe. The shell indices in these two species may overlap (Table 1), but their mean values are significantly different (t- test, Р < 0.01). Pisidium simplex is identified by an extremely extended pointed apex. The best distinction is provided by the elongation index (H/L) and hinge index (hh/H). The pattern of surface sculpture in this species is similar to that in P moitessierianum, but in most of the studied populations the ribs were less pro- nounced. Pisidium simplex differs from P. punc- tiferum, as described by Guppy (1867) and Burch (1975), in having a smaller shell with markedly asymmetrical umbones and less pronounced sculpture (Figs. 6, 8), but resem- bles the latter in the shape of the nephridium and arrangement of mantle muscles (Figs. 7, 9). DISCUSSION The presence of concentric umbonal folds around the umbones is a common feature shared by Р. moitessierianum and the nomi- TAXONOMIC REVISION OF PISIDIUM PUNCTATUM 343 FIG. 4. Anatomy of Pisidium moitessierianum. Western Lake Erie, Michigan. A, gross anatomy; B, gills; C, mantle edge; D, nephridia. Scale bars = 1 mm for A-C and 0.5 mm for D. native form of P. punctatum, as mentioned by Kuiper (1962b). Identity of these taxa is also supported by the greatly simplified dorsal lobe of nephridium and weak mantle muscles. The present data agree well with published de- scriptions of the nephridium in P. moitessieri- anum (Odhner, 1923; Korniushin, 1996, 1998); this form of the dorsal lobe never oc- curs in the other species of Pisidium (Kor- niushin, 1996). Both North American and European popula- tions of P. moitessierianum have all the char- acters described by Odhner (1921, 1923), Kuiper (1962a), and Meier-Brook (1970) for the subgenus Neopisidium. These features in- clude an enclosed, but not introverted liga- ment, presence of one demibranch and one siphon, and an upper, dorsal position of the brood pouch. A markedly broadened hinge with arched cardinal teeth is reported in all European de- scriptions of P. moitessierianum (Ellis, 1978; Piechocki, 1989; Glóer & Meier-Brook, 1998). However, as shown here, in some European lacustrine populations, the hinge may be markedly weakened. Thus, while having some differences from the typical form, P punctatum cannot be distinguished as a geographic vari- ety or subspecies of P. moitessierianum at the present time. The known North American distribution of PR moitessierianum (Sterki, 1895, as P. puncta- tum s. str. and P. punctatum var. armatum; Kuiper, 1962b; Grigorovich et al., 2000) is lim- ited to the lower Great Lakes basin. It should be noted that four other sphaeriid species that have been recognized as introductions from Europe, are also restricted to habitats within the Great Lakes and/or neighboring drainages (Herrington, 1962; Burch, 1975; Clarke, 1981). Thus, the introduced origin of P moitessierianum from Europe seems likely. However, until further evidence as to its origin and distribution on the continent is obtained, this species should be considered as crypto- genic, according to definition of Carlton (1996). Species distinctness of P simplex con- cluded from differences in shell outline, form of hinge teeth, arrangement of the mantle muscles and form of the nephridium, can fur- ther be confirmed by statistically significant differences in shell indices. In contrast to P moitessierianum, P. simplex has a much broader distribution in North America, from the Pacific to the Atlantic coast. Affinity be- tween P. simplex and P. punctiferum (Guppy, 1867) is supported by the similarity in their anatomy, particularly in their mantle muscula- ture and nephidium characteristics. These species are probably closely related, despite the geographic gap separating their ranges (Burch, 1975). Thus, southern, neotropical affinities of P. simplex are assumed. Pisidium punctiferum is traditionally in- cluded in the subgenus Neopisidium, along- side P. moitessierianum. However, the dorsal lobe of its nephridium, like that of P. simplex, is more similar to that of Afropisidium species reported by Korniushin (1998), than to P moitessierianum. Affinity of P. simplex to the subgenus Afropisidium is also suggested by the slightly elevated ligament. The latter sub- genus is recognized as a South American na- tive (Kuiper, 1962a). However, a decision on the subgeneric arrangement of P. punctiferum and, consequently, P. simplex, would be pos- sible only after thorough anatomical investi- KORNIUSHIN, GRIGOROVICH 8 MACKIE 344 Хэ} ay] ul UBAIH эле siaquinu anbojejeo *, цим pajeo -IPUI $! апаице o oads jo uoenesqo ON “ajeudoldde элэцм uonelAep рлерие}$ pue иеэш '(sesayjuaJed ul) зи ‘(N) azis ajdwes Jussaıdaı siaquiny ‘WWW и! эле зио!зиацир ||V :310N (Gt'0-8£ 0) (r20-220) (950-150) (Zr0°0-EE0'0) (620-890) (68 0-28'0) rE00 + 27'0 8000 = 520 ZI00++SO +000 + 900 — 590`0 = 2/0 91007480 (1€'0-£€'0) (12:0-61'0) (090-950) (250'0-€b0'0) (180-220) (98'0-08'0) 9100 = SEO 60007020 ¿I00+8S0 #00`0 + $#0`0 SS00+8Z0 12007+€80 (9€°0-SE'0) (02:0-61'0) (650-2590) (650'0-9p0'0) (280-820) (58`0-28`0) (85`0-65`0) (020-410) (090-950) (Ssto'0-8€0'0) (18`0-6/`0) (£8:0-08'0) £L0OO+/E0 LLOO0+8r0 +blOO+8S0 €000+Zb00 2200+6/0 8000 + 280 ce 0 6L'0 950 770`0 (18'0-98'0) (s8:0-€8'0) (er0-LE'0) (02°0-$1'0) (850-590) (650 0-+£0'0) (28'0-7/`0) (Р8`0-08`0) 950'0 + SO 7100 +110 81070 + 459`0 8000 + EPOO 22007820 2100 + E80 (190`0-9$0`0) (18'0-6/`0) (£6 0-GS8'0) 4 mi } 9000 +SS00 8200 + 8/0 S200 + 060 (320'0-690'0) (08 0-G/'0) (16'0-18`0) у s 2 Z00'0 + 140'0 1Z2000+ ¿40 2100 +060 (220'0-850'0) (220-290) (26'0-98'0) 7 z Z 600`0 + £90 0 LEO'O = 120 1Z0'0 + 88`0 : я a (£90'0-650'0) (6/`0-7/`0) (68`0-88`0) (27'0-0$`0) (y2:0-07'0) (£90-090) (290'0-€b0'0) (£E8°0-r2'0) (68°0-S8'0) sooo + 17'0 LIOO0O+2Z0 HOOFLIO LIO'O + /S00 ErOOFSLO S007 2/80 €t'0 02'0 650 ss0 0 (22'0-89'0) (16'0-06'0) 7/dn Vs Vy H/uu H/M VH saJoIpul suolsuswip O9IXEN ‘шпладоипа y 11819 IS aye] Xaydwıs y aye] JaAeag ‘xa/dwis y spunoyy ueipu| ‘xa/dwis y JOAIH x0- ‘sadAjuAs ‘ха!аши$ en winjejound y 49919 ч$еэ^л ‘sadAjuAs ‘xa/duis “Jen wnjejound y aÁ0|9g 9-87] ‘шпивива!55аиош y лам ‘WNUBLAISSA}IOW y 9113 aye] Wa}sem ‘шпивие!5$аиош y 11819 IS эхел ‘wNueUaIssayioW y sadÁAjuÁs ‘штвиие Jen штырипа 4 sadAjuÁs ‘wnjejound 4 sam eso] pue saidads зациеэс| оцаелбоэб jualayip Wood) Saioeds WINIpISig JO зэцзиэовелецо OUJaWOYdoW ‘| 37191 FIG. 5. Shells of Pisidium punctatum var. simplex. A, Lectotype, CMNH 2804 (external frontal and lateral and internal lateral view of left valve and hinge features of single individual). Lillycash Creek, Illinois; B, CMNH 272 (external frontal and lateral view and hinge of left valve). Fox River, Wisconsin; C, CMNH 271 (external frontal and lateral view of left valve and hinge features of single individual). New York State; D, CMNH 10563 (external frontal and lateral view and hinge of left valve), Beaver Lake, Washington Sate; E, CMNH 10399 (external frontal and lateral view and hinge of left valve). Lake Ontario, Ontario. Scale bar = 1 mm. FIG. 6. Shell and hinge morphology of Pisidium simplex. Paralectotype, CMNH 3781. Brook near Indian mounds, Michigan: A, shell; B, external shell sculpture; C, hinge. St. Clair River delta, Ontario: D, shell; E, hinge and internal shell surface. Scale bars = 0.1 mm. 346 KORNIUSHIN, GRIGOROVICH 8 MACKIE FIG. 7. Anatomy of Pisidium simplex. St. Clair River delta, Ontario. A, gross anatomy; B, gills; C, mantle edge; D, nephridia. Scale bars = 1 mm. FIG. 8. Shell and hinge morphology of Pisidium punctiferum. San Diegito, Mexico, ex. A. C. Hinkley, FMNH 59511. A, shell; B, hinge. Scale bars = 1 mm. gation of the South American material, which is beyond the scope of this paper. ACKNOWLEDGEMENTS We gratefully appreciate A. Baldinger, J. E. Rawlins, R. L. Davidson and C. Sturm for al- lowing us to study Pisidium collections at their institutions, C. Meier-Brook for providing alco- hol material of P. punctiferum, Dr. E. Wasch for conducting S.E.M., and V. Heinrich for pro- cessing photographs. Comments by G. M. Davis, E. V. Coan, H. J. Maclsaac, R. |. Co- FIG. 9. Anatomy of Pisidium punctiferum. Guade- loupe, collection of C. Meier-Brook. A, mantle edge; B, nephridia. lautti, S. A. Bandoni, D. Banninga, and C. Matkovié improved the clarity of manuscript. This study was supported by the Collection Study Grant of the AMNH, Bass Fellowship and A. von Humboldt fellowship to AVK, and by NATO and GLIER postdoctoral fellowships to IAG. LITERATURE CITED BURCH, J. B., 1975, Freshwater sphaeriacean clams (Mollusca: Pelecypoda) of North America. Malacological Publications, Hamburg, Michigan. 96 pp. TAXONOMIC REVISION OF PISIDIUM PUNCTATUM 347 CARLTON, J. T., 1996, Biological invasions and cryptogenic species. Ecology, 77: 1653-1655. CLARKE, A. H., 1981, The freshwater molluscs of Canada. National Museums of Canada, Ottawa. 446 pp. D'ITRI, F. M., ed., 1997, Zebra mussels and aquatic nuisance species. Ann Arbor Press, Chelsea, Michigan. 638 pp. ELLIS, A. E., 1978, British freshwater bivalve Mol- lusca. Synopses of the British fauna, (New Se- ries)11. The Linnean Society of London. 91 pp. FAVRE, J., 1943, Revision des especes de Pisid- ium de la collection Bourguignat du Museum d'Histoire Naturelle de Geneve. Revue Suisse de Zoologie, Suppl. 50: 1-64. GLOER, P. & C. MEIER-BROOK, 1998, Súss- wassermollusken. Ein Bestimmungsschlussel fur die Bundesrepublik Deutschland. Deutscher Ju- genbund für Naturbeobachtung, Hamburg. 136 рр. GRIGOROVICH, I. A., KORNIUSHIN, А. V. & Н. J. MACISAAC, 2000, Moitessier's pea clam Pisid- ium moitessierianum (Bivalvia, Sphaeriidae): a cryptogenic mollusc in the Great Lakes. Hydrobi- ologia, 435: 153-165. GUPPY, R. J. L., 1876, Description of a new fresh- water bivalve found in Trinidad. Annals and Mag- azine of Natural History, (3) 19: 160-161. HERRINGTON, H. B., 1954, Pisidium species and synonyms, North America, north of Mexico, The Nautilus, 67: 131-138. HERRINGTON, H. B., 1962, A revision of the Sphaeriidae of North America (Mollusca: Pelecy- poda). Miscellaneous Publications, Museum of Zoology, University of Michigan, 118: 1-74, pls. 1-7. KENNARD, A. S. & B. B. WOODWARD, 1926, Syn- onymy of the British non-marine Mollusca (Re- cent and post- Tertiary). The British Museum (Nat- ural History), London. 447 pp. KORNIUSHIN, A. V., 1996, Bivalve molluscs of the superfamily Pisidioidea in the Palaearctic region: fauna, systematics, phylogeny. Schmalhausen Institute of Zoology, Kiev, Ukraine. 176 pp. (In Russian). KORNIUSHIN, A. V., 1998, A comparative investi- gation of nephridia in fingernail and pill clams. Bi- valvia |. Malacological Review, Suppl., 7: 53-63. KUIPER, J. G. J., 1962a, Note sur la systématique des pisidies. Journal de Conchyliologie, 102: 53- 572 KUIPER, J. G. J., 1962b, Systematische Stellung and geographische Verbreitung von Pisidium tenuilineatum. Archiv fúr Molluskenkunde, 91: 173-181. MACKIE, С. L., D. $. WHITE & T. W. ZDEBA, 1980, A guide to freshwater molluscs of the Laurentian Great Lakes with special emphasis on the genus Pisidium. Environmental Research Laboratory, Office of Research and Development U.S. Envi- ronmental Protection Agency, Duluth, Minnesota. 144 pp. MEIER-BROOK, C., 1970, Untersuchungen zur Bi- ologie einiger Pisidium-Arten (Mollusca; Eulamel- libranchiata; Sphaeriidae). Archiv fúr Hydrobiolo- gie, Suppl., 38: 73-150. NALEPA, T. F. & D. W. SCHLOESSER, eds., 1993, Zebra mussels: biology, impact and control. Lewis Publishers, Boca Raton, Florida. 810 pp. ODHNER, N., 1921, On the some species of Pisid- ¡um in the Swedish State Museum. Journal of Conchology, 16: 218-223. ODHNER, N., 1923, On the anatomical character- istics of some British pisidia. Proceedings of the Malacological Society of London, 15: 155-161. PALADILHE, M., 1866, Nouvelles miscelaneés malacologiques. |. Especes inédites, nouvelles ou peu connues du département de L'Hérault. Revue et Magasin de Zoologie, (2) 18: 168-174. PIECHOCKI, A., 1989, The Sphaeriidae of Poland (Bivalvia: Eulamellibranchiata). Annales Zoo- logici, 42: 249-320. STERKI, V., 1895, Two new pisidia, The Nautilus, 8: 97-100. STERKI, V., 1905, New varieties of the North Amer- ican pisidia. The Nautilus, 19: 80-84. Revised ms. accepted 1 January 2001 MALACOLOGIA, 2001, 43(1-2): 349-365 INDEX Taxa in bold are new; pages in italic Cecilioides 38, 47, 53 indicate figures of taxa. abyssicola, Cadlina 238, 240-241, 242- 244, 243, 307, 309 abyssorum, Bathydoris 237 Achatina fulica 57 gundlachi 47 octona 46 pulchella 44 Achatinidae 224-225, 227, 232-233 achroma, Discodoris 240, 274, 275-276, PT ЗО acicula, Buccinum 47 Caecilianella 47 Cecilioides 38, 47 Adrihydrobia 162, 164, 168 Adriolitorea 134, 165 affinis, Cadlina 243 Afropisidium 343 albanica, Horatia 130-131, 175 albobrunnea, Dendrodoris 306 alexandrina, Biomphalaria 225, 232-234 algesirae, Siphonaria 225, 232-234 Allopeas 45 gracile 38, 45, 54, 57 micra 38, 45, 54, 57 alte, Laevicaulis 225, 232-234 Alzoniella 162, 164, 169-170 amanda, Mathilda 328, 329, 331-334 Amblema 94 Ambleminae 89 amidicus, Sheitanok 175, 179 Amnicola globulus 175 Amnicolidae 168 Amnicolinae 168 amnicum, Pisidium 337 Ampullariidae 13 anatolica, Islamia 175, 178 Ancylidae 16 Aneita 233 ankyra, Rostanga 278, 294, 295-296, 296 Anodonta 94 cataracta 94 anti, Hadziella 121, 122, 175, 178 antillarum, Leptinaria 46 Tornatellina 46 aperta, Caecilianella 47-48 Cecilioides (Geostilbia) 47 Macrospira 47-48 apiculata, Quadrula 89-90, 98 Aplysia 225, 227, 232-234 araneosa, Paradoris 257, 281, 282-283, 284-285, 287-288, 309 Archidoris 252, 255, 260 australis 237 kerguelenensis 244 rubescens 244 Architectonicoidea 327-328 Arganiella 106, 108, 158, 162-163, 165- 166, 169-170, 773, 209 pescei 106-108, 107, 110, 176, 178 Arion lusitanicus 25-31, 28-29 fasciatus 25, 30 hortensis 30 intermedius 29-30 subfuscus 25, 30 ascendens, Opeas 46, 53 asiana, Daudebardiella 175, 178 aspersa, Helix 9, 225, 232-234 Asteronotus 256 ateni, Islamia 175, 178, 182, 184 Microna 175 Neohoratia 103, 177 Athoracophoridae 223-224, 227, 232-233 Athoracophorus 227 bitentaculatus 223-225, 232-234 atromaculata, Peltodoris 277, 281 aureola, Mandarina 9 auricularia, Lymnaea 225, 232-234 auris, Voluta 43 aurissileni, Plekocheilus 43 Plekocheilus (Plekocheilus) 43 australis, Archidoris 237 Austrodoris 237, 244, 252, 255, 260, 307-308 caeca 240, 248, 249-251, 252, 255, 309 georgiensis 244, 248 kerguelenensis 237, 240, 244, 246-247, 248, 252, 255, 308 laboutei 240, 252, 253-254, 255, 309 macmurdensis 248 Avenionia 162, 164, 169-170 Axinodon symmetros 324 azarum, Islamia 175, 178 Neohoratia 175, 177 349 350 Baicalia 75, 81-82 carinata 60, 62-65, 68, 71, 73, 75, 76, 80-82 carinatocostata 60, 76 dybowskiana 60, 64-65, 67, 68, 71, 75, 76, 80-82 turriformis 60, 65, 66, 68, 75, 81 Baicaliidae 59, 77, 83 Baicaliinae 59 Bakerilymnaea cubensis 225, 232-234 balcanica, Pseudoislamia 152, 175, 179 Balea biplicata 225, 232-234 Baptodoris 237, 299, 302, 307 cinnabarina 299, 302 mimetica 302 phinei 278, 299, 300-301, 302, 308-309 barbadensis, Bulimus 42-43 Carocolla 50 Gastrocopta 38, 41, 57 Glyphyalinia 38, 48-49, 54 Helicina 39 Helix 50 Helix (Carocolla) 50 Helix (Dentellaria) 50 Lacteoluna selenina 50 Lucidella 33, 37-38, 40, 53-54 Lucidella (Poeniella) 39-40 Pseudopineria 33, 37-38, 45, 53-54 Pupa 41-42 Succinea 38, 41, 57 Truncatella 38, 40, 57 Bathydoris 237 abyssorum 237 beckiana, Opeas 46 Stenogyra 45 Beckianum beckianum 38, 45, 54, 57 beckianum, Beckianum 38, 45, 54, 57 beckianus, Bulimus 45 Belgrandia 162, 164, 169-170 Belgrandiella 162, 164, 169-170 bermudensis, Succinea 38, 41 bendidis, Islamia 175, 179 bicolor, Gulella 38, 53 Ennaea (Huttonella) 48 Gulella (Huttonella) 48 Pupa 48 bidentata, Mysella 323-324 bilabiata, Truncatella 40 Truncatella (Truncatella) 40 Biomphalaria alexandrina 225, 232-234 glabrata 225, 232-234 peregrina 16 INDEX biplicata, Balea 225, 232-234 birsteini, Horatia 175, 178 bitentaculatus, Athoracophorus 223-225, 232-234 boholiensis, Discodoris 271 Boreodoris setidens 295 borutzkii, Horatia 175, 178 bosniaca, Dabriana 109, 175, 178 Islamia 175, 178 boui, Fissuria 110, 112, 115-116, 118, 175, 178, 195, 198 bourguignati, Valvata 172, 175, 202 Bracenica 106, 108-109, 158, 162-163, 165-166, 168, 173 spiridoni 108-109, 176, 178 Brachypodella costata 36, 38, 44, 57 tatei 45 Brachypus 44 costatus 44 Bradybaena similaris 38, 52, 57 Bradybaenidae 52 brezicensis, Kerkia 138, 175, 179 brodieae, Dendrodoris 257, 304, 305-306, 306 brusinae, Horatia 175 Pseudohoratia 152, 175, 179 Buccinum acicula 47 zebra 44 bugensis, Dreissena 337 bulbosum, Pleurobema 87-90, 91, 92-94, 94, 97-98, 97, 99 Bulimulus 43 (Bulimulus) diaphanus fraterculus 43 (Bulimulus) fuscus 43 diaphanus 43 exiles 42 exilis 42 fraterculus 43 fuscus 33, 37-38, 42-43 guadalupensis 38, 42-43, 54, 57 tenuissimus 38, 43 (Thaumastus) exilis 42 Bulimus barbadensis 42-43 beckianus 45 (Borus) oblongus 49 caraccasensis 45 diaphanus 43 exilis 42 fraterculus 43 fuscus 42, 53-54 goodalli 46 gracile 45 guadalupensis 42 gundlachi 47 oblongus 49 octonoides 45 octonus 46 pumilus 46 subula 45 viequensis 45 burnabasa, Horatia 175 Islamia 175, 178 Bythinella 162, 164, 168, 169-170 bythiniopsis, Maackia 64-65, 67, 68, 71, 71, 76, 80-83 Maackia (Eubaicalia) 60 Bythiospeum 162, 164, 169-170 Cadlina 237-238, 242, 244, 245, 307-308 abyssicola 238, 240-241, 242-244, 243, 307, 309 affinis 243 dubia 242 georgiensis 243-244 kerguelensis 243 laevis 238 magellanica 243 nigrobranchiata 242 sparsa 243 willani 242 caeca, Austrodoris 240, 248, 249-251, 252. 255, 309 Caecilianella acicula 47 aperta 47-48 (Caecilioides) consobrinus minutissima 48 consobrinus minutissima 47 gundlachi 47 minutissima 47-48 Caecilioides minutissima 47 callosa, Goslineria 256, 257-260 Camaenidae 50 canaliculata, Pomacea 13-22, 18-19, 21 capensis, Diaulula 289, 291 caraccasensis, Bulimus 45 Stenogyra 45 carinata, Baicalia 60, 62-65, 68, 71, 73, 75, 76, 80-82 carinatocostata, Baicalia 60, 76 cariosa, Lampsilis 94 Carocolla barbadensis 50 carolinensis, Glyphyalinia 49 Retinella (Glyphyalinia) 49 INDEX 351 caroliniana, Uniomerus 90, 92-93, 96, 97 casertanum, Pisidium 337 cataracta, Anodonta 94 Cecilioides acicula 38, 47 aperta 38, 47, 53 (Geostilbia) aperta 47 (Karolus) consobrinus minutissima 47 celtica, Onchidella 225, 232-234 Cepaea hortensis 9 nemoralis 9 cerebralis, Diaulula 278, 291, 292-293, 294 cerevisiae, Sacharomyces 221 Cerion 42 uva 38, 42 Cerionidae 42 Chaetoceros gracilis 315 Chilina parchappii 16 Chilinidae 16 Choanomphalus 65 Chrysallida 328 cianensis, Islamia 175, 178, 202 ciliata, Teratobaikalia 64-65, 67, 68, 71, 72, 76, 79-83 Teratobaikalia (Trichiobaikalia) 60 cinnabarina, Baptodoris 299, 302 Clameia 162, 164, 168 clandestina, Teratobaikalia nana 68, 73, 76, 79-80, 82 Teratobaikalia (Baicaliella) nana 60 Clausiliidae 224 clava, Pleurobema 87-90, 92, 96, 98 clavigera, Thais 224-226, 232-234 coccinea, Doris 294 Cochliopinae 168 columella, Godlewskia 68 complanata, Doris 268 Elliptio 90, 94 compressum, Pisidium 340 concentrica, Gundlachia 16 conoidea, Helicina 37, 39-40 consolationis, Hauffenia minuta 205 Horatia minuta 173, 175 Islamia 103, 155, 175, 178, 184, 199- 201. 201, 205, 206, 207 contabulata, Pseudobaikalia 65, 76, 80, 82 Pseudobaikalia (Pseudobaikalia) 60 convexa, Helicina 37 Corambidae 237 Corbicula fluminea 87, 337 corneum, Sphaerium 337 Cornirostridae 328 352 coronadoi, Islamia 175, 179 Neohoratia 177 Valvata 175 coronata, Dendrodoris 304 costata, Brachypodella 36, 38, 44, 57 Cylindrella 44 Cylindrella (Gongylostoma) 44 Maackia 65, 71, 80-81 Maackia (Maackia) 60 Siphonostoma 44 costatus, Brachypus 44 crassidens, Elliptio 90, 98 crawfordi, Discodoris 271 Cristalabrum primum 9 cristatum, Pisidium lilljeborgi 340 cruciatum, Pisidium 340 cubensis, Bakerilymnaea 225, 232-234 cuneata, Mysella 323 Cylindrella costata 44 (Gongylostoma) costata 44 Dabriana 109, 111, 158, 162-163, 165-166, 168, 173 bosniaca 109, 175, 178 Dalmatella 111, 158, 162-163, 165-166, 168, 173 miljackae 176 sketi 111, 176, 178 danubialis, Hauffenia 140, 175, 178, 180 Lobaunia 103, 138, 175, 180 Daphniola 106, 111, 113-114, 150, 158, 162-163, 165-166, 168, 173 exigua 107, 110, 111 exigua exigua 175, 178 exigua pangaea 176, 178 graeca 111, 113, 175 hellenica 113 Daudebardiella 165 asiana 175, 178 naegelei 176, 178 decolorata, Happiella 37-38, 49, 57 Zonites 49 deformis, Helix 48 Streptaxis 48 deminuta, Hadziella 120, 122, 175, 178 Hadziella ephippiostoma 175 Dendrodoris 237, 302, 304, 305, 306-308 albobrunnea 306 brodieae 257, 304, 305-306, 306 coronata 304 denisoni 306 INDEX guttata 306 lugubris 302 nigra 306 orbicularis 278, 302, 303, 304, 309 denisoni, Dendrodoris 306 Dentellaria isabella 51 dentiens, Helix 50-51 Helix (Helicodonta) 51 Pleurodonte 38, 51 depressa, Ohridohauffenia 144, 175, 179 Pseudamnicola 105, 144, 175 Deroceras laeve 26 reticulatum 25-26, 31 diaphanus, Bulimulus 43 Bulimus 43 diaphora, Pharodoris 240, 257, 260, 261- 263, 264, 266, 307, 309 Diaulula 237, 288-289, 291-292, 294, 307 capensis 289, 291 cerebralis 278, 291, 292-293, 294 immaculata 278, 288, 289-290, 291, 294 morosa 289, 291 rubra 289 sandiegensis 291, 294 dioscoricola, Helix 42 Pupisoma 53 Pupisoma (Ptychopatula) 42 Discodoris 237, 240, 271, 272-273, 274, 277, 281, 507 achroma 240, 274, 275-276, 277, 307 boholiensis 271 crawfordi 271 palma 271 Discohelicidae 328 Dolapia 106, 136, 165 ornata 135 Doris coccinea 294 complanata 268 hispida 291 repanda 238 sandiegensis 288-289 vestita 291 dorsalis, Xylophaga 323 Dreissena bugensis 337 polymorpha 87, 337 Dreissenidae 313 drimica, Ohridohauffenia 145, 175, 179 Pseudamnicola (Rotondia) 175 dubia, Cadlina 242 dubiosa, Parabaikalia elata 60, 71, 76, 80-82 duthiersii, Teratobaikalia 65, 68, 71, 72, 76, 80-83 Teratobaikalia (Trichiobaikalia) 60 dybowskiana, Baicalia 60, 64-65, 67, 68, 71,75, 76, 80-82 Dybowskiola 82 edlaueri, Hauffenia 171, 175, 178 Horatia (Hauffenia) 175 elata, Parabaikalia 63, 79 Parabaikalia elata 60, 64-65, 68, 71, 76, 79-82 Elliptio 89-90, 94, 96, 97 complanata 90, 94 crassidens 90, 98 Ennaea (Huttonella) bicolor 48 ephippiostoma, Haaziella 110, 119, 120, 122, 175, 178 epirana, Horatia (Neohoratia) 175 Islamia 175, 179 Neohoratia 171-172 Erycina 323 erythropomatia, Erythropomatiana 114, 161, 165, 180 Hauffenia 103, 110, 112, 117-118, 171, 175, 178, 180, 189 Valvata 103-104, 114-115, 175, 180 Erythropomatiana 103-104, 114-115, 127, 139, 161-163, 165, 168-171, 180 erythropomatia 114, 161, 165, 180 verdica 115, 140, 141, 143-144, 161, 177,189 Eubaicalia 81 Eulota similaris 52 exigua, Daphniola 107, 110, 111 Daphniola exigua 175, 178 Horatia (Daphniola) 113 Horatia (Horatia) 113 Valvata 111, 113, 175 exilis, Bulimulus 42 Bulimulus (Thaumastus) 42 Bulimus 42 Heraultia 155, 175, 178, 184, 208, 208, 210 Horatia 103 Valvata 108, 172-175, 200, 208-209 fasciata, Helicina 39 Helicina fasciata 37 fasciatus, Arion 25, 30 fellowski, Peltodoris 281, 284 fezi, Islamia 175, 179 INDEX 353 Neohoratia 177 Valvata (Tropidina) 175 Fissuria 103, 115-116, 158, 162-163, 165- 166, 169-170, 174 boui 110, 112, 115-116, 118, 175, 178, 195, 198 planospira 116, 176, 178 raehlei 155, 172, 176, 178, 193, 194 flava, Fusconaia 90, 92, 98 florii, Parabaikalia 62-65, 68, 71, 74, 79 Parabaikalia florii 60, 64-65, 68, 71, 76, 79-82 fluminea, Corbicula 87, 337 fontinalis, Horatia 130-131, 175 Fossaria truncatula 225, 232-234 fraterculus, Bulimulus 43 Bulimulus diaphanus 38, 43 Bulimulus (Bulimulus diaphanus) 43 Bulimus 43 Helix (Cochlogena) 43 Fruticicola similaris 52 fulica, Achatina 57 Fusconaia 92 flava 90, 92, 98 fuscus, Bulimulus 33, 37-38, 42-43 Bulimulus (Bulimulus) 43 Bulimus 42, 53-54 gaillardoti, Islamia 175, 179 Mienisiella 134 Valvata (Cincinna) 175 galbanus, Platydoris 284 Galeommatoidea 313 gaiteri, Islamia 175, 179, 202 Gastrocopta barbadensis 38, 41, 57 pellucida 38, 42 gasulli, Hauffenia (Neohoratia) 175 Horatia 175, 178 Neohoratia 177 gatoa, Horatia 174-175, 178 georgiensis, Austrodoris 244, 248 Cadlina 243-244 Geitodoris 237, 268-269, 271, 307-308 lutea 271 ohshimai 271 pallida 240, 260, 268-269, 269-270, 271 pusae 271 sticta 271 gjorgjevici, Lyhnidia 141, 175, 179 Ohridohoratia (Ohridohauffenia) 105, 144-145, 175 354 Ohridohoratia 144 glaber, Streptaxis 34, 38, 54, 57 Streptaxis (Odontertemon) 48 Streptaxis (Streptartemon) 48 glabra, Litigiella 323 Lymnaea 225, 227, 232-234 Streptaxis 48 glabrata, Biomphalaria 225, 232-234 globulina, Horatia minuta 173 Islamia 103, 175-177, 179, 184, 195, 198, 199-203, 201, 204 Neohoratia 198 Valvata 172-173, 175, 199-200, 202-203 globulus, Amnicola 175 Islamia globulus 175, 179, 183-184 Neohoratia 177 Neohoratia globulus 103, 177 Glyphyalinia 49 barbadensis 38, 48-49, 54 carolinensis 49 Gocea 116, 120, 158, 162-163, 165-166, 169-170, 173 ohridana 116, 118-120, 119, 176, 178 Godlewskia 81 columella 68 pulchella 60, 64-65, 68, 71, 76, 79-82 goodalli, Bulimus 46 Opeas 46, 53 Stenogyra 46 Goslineria 237, 256, 307 callosa 256, 257-260 gracile, Allopeas 38, 45, 54, 57 Bulimus 45 gracilis, Chaetoceros 315 graeca, Daphniola 111, 113, 175 Islamia 175, 179 Graecoanatolica 162, 164, 168 granulata, Paradoris 281 Graziana 162, 164, 169-170 guadalupensis, Bulimulus 38, 42-43, 54, 57 Bulimus 42 Gulella bicolor 38, 53 (Huttonella) bicolor 48 Gundlachia concentrica 16 gundlachi, Achatina 47 Bulimus 47 Caecilianella 47 Opeas 47 Stenogyra 47 guttata, Dendrodoris 306 INDEX hadei, Hauffenia 175, 178 Horatia (Neohoratia) 175 Neohoratia 171 Haaziella 120, 122, 158, 158, 162-163, 165-166, 169-170, 173 anti 121, 122, 175, 178 deminuta 122, 175, 178 ephippiostoma 110, 119, 120, 122, 175, 178 ephippiostoma deminuta 175 krkae 122, 176, 178 rudnicae 122, 176, 178 sketi 122, 176, 178 thermalis 122, 177-178 umbilicata 177 haazii, Lyhnidia 139, 140, 141, 175, 179 Halgerda 238 hannense, Opeas 38, 45-46, 53, 57 hannensis, Hélice (Cochlicelle) 46 Helix 46 Happiella decolorata 37-38, 49, 57 Hauffenia 103-104, 115, 122, 124-127, 134, 136, 139, 144, 154, 156-157, 158, 159, 161-163, 165-166, 168-181, 171, 177, 179-181, 181, 186, 188-190, 192-193, 199 danubialis 140, 175, 178, 180 edlaueri 171, 175, 178 erythropomatia 103, 110, 112, 117-118, 171179178, 1804189 hadei 175, 178 (Hauffenia) subcarinata 177, 188 (Hauffenia) tovunica 177, 190 jadertina 171, 175, 178 jadertina sinjana 176 kerschneri 103, 171, 175-176, 178, 180- 181, 185, 186 kerschnen loichiana 176, 181-182, 185-186 kerschneri kerschneri 182, 186 kusceri 136, 176 lucidula 171-172, 176, 178 media 103, 171, 176, 178, 180, 186, 187, 192 michleri 103, 122, 123, 124-127, 128- 129, 176, 189-190 minuta 173, 198 minuta consolationis 205 minuta spirata 176, 203 Neohoratia 173 (Neohoratia) coronadoi schuelei 176 (Neohoratia) gasulli 175 (Neohoratia) minuta 173 plana 171, 176, 178 raehlei 171-172 sinjana 171, 176, 178 solitaria 171, 176 subcarinata 103, 128, 171, 177-178, 180, 188, 191 subpiscinalis 103-104, 115, 117-118, 128-129, 140, 142, 144, 161, 171, 177- 178, 180, 189 tellinii 103, 110, 117-118, 122, 123-124, 127, 128-129, 161, 165, 171, 176-178, 180, 189-190, 192 tovunica 103, 161, 171, 177-178, 180, 190, 191, 192 verdica 189 (Vrania) 165 wagneri 103, 117, 128, 151, 155, 171, 177-178, 180, 186, 188, 192 wienerwaldensis 103, 171, 177-178, 180, 191, 192 Heleobia 163-164, 168-170 parchappii 16 Hélice (Cochlicelle) hannensis 46 (Hélicelle) similaris 52 Helicidae 1, 224-225, 227, 232-233 Helicina 37, 40 barbadensis 39 conoidea 37, 39-40 convexa 37 fasciata 39 fasciata fasciata 37 fasciata substriata 36-39, 57 occidentalis 37 plicatula 40 rugosa 40 substriata 37, 39 Helicinidae 37, 39 Helicogena isabella 50 Helix 52 (Achatina) micra 45 aspersa 9, 225, 232-234 barbadensis 50 (Carocolla) barbadensis 50 (Cochlogena) fraterculus 43 deformis 48 (Dentellaria) barbadensis 50 (Dentellaria) isabella 51 (Dentellaria) perplexa 51 dentiens 50-51 dioscoricola 42 (Dorcasia) similaris 52 INDEX 355 (Fruticicola) similaris 52 hannensis 46 (Helicodonta) dentiens 51 (Helicodonta) perplexa 51 incisa 48-49, 54 isabella 50-51 (Microphysa) turbiniformis 50 (Microphysa) vortex 50 oblongus 49 perplexa 51 provisoria 51-52 selenina 50 similaris 52 subaquila 50 turbiniformis 50 vortex 50 hellenica, Valvata (Cincinna) 111, 113, 175 Daphniola 113 henslowanum, Pisidium 337 Heraultia 103, 158, 162-163, 165-166, 169-170, 174, 174, 207, 209 exilis 155, 175, 178, 184, 208, 208, 210 herderiana, Maackia (Eubaicalia) 59-60 Maackia 62-65, 68, 70, 71, 76-83 hermaphrodita, Phasmarhabditis 30 hispida, Doris 291 holoserica, Lucidella 40 Hoplodoris 256 Horatia 103-104, 113, 122, 127, 130-131, 141, 144, 150, 158, 162-163, 165-166, 169-170, 173, 173-174, 179, 209 albanica 130-131, 175 birsteini 175, 178 borutzkii 175, 178 brusinae 175 burnabasa 175 (Daphniola) exigua 113 (Daphniola) exigua pangaea 113-114, 176 erythropomatia kerschneri 175, 181 exilis 103 fontinalis 130-131, 175 gasulli 175, 178 gatoa 174-175, 178 (Hauffenia) 127, 165, 175 (Hauffenia) edlaueri 175 (Hauffenia) lucidula 176 (Hauffenia) lucidulus 176 (Hauffenia) raehlei 103, 116, 176, 193, 195 (Hauffenia) stankovici 147, 177 356 (Hauffenia) tellinii 103, 122, 177, 189-190 (Hauffenia) valvataeformis 122, 177, 189- 190 (Horatia) exigua 113 klecakiana 127, 128, 130, 130-131, 175- 178 knorn 178 knorri 131, 175 kusceri 156, 176 letourneuxi 130-131, 176 ljovuschkini 176, 178 macedonica 131, 176, 178 minuta 173 minuta consolationis 173, 175 minuta globulina 173 minuta minuta 173 minuta spirata 173 (Neohoratia) epirana 175 (Neohoratia) hadei 175 novoselensis 131, 176, 178 obliqua 130-131, 176 obtusa 130-131, 176 Ohridohauffenia (Neohoratia) 165 palustris 130-131, 176 parvula 176, 178 polinskii 152, 176 praeclara 130-131, 176 servaini 131, 133-134, 176 sokolovi 176, 178 st. zaumi 177 sturmi 174, 177-178 supracarinata 177, 188 tellinii 103-104, 144 valvataeformis 103, 126 verlikana 130-131, 177 hortensis, Arion 30 Cepaea 9 humerosa, Teratobaikalia nana 68, 73, 82 Teratobaikalia (Baicaliella) nana 60 Hyalina incisa 34, 48 Hyalogyrinidae 328 Hydrobia 159, 163-164, 168-170 valvataeformis 131, 133-134, 177 Hydrobiidae 16, 59, 103, 111 Hydrobiinae 168 Iglica 163-164, 168 immaculata, Diaulula 278, 288, 289-290, 291, 294 imperfecta, Paradoris 257, 285, 286-287, 287-288 INDEX implicans, Miradiscops 38, 49, 53 Vitrea 49 Zonites 49 incisa, Helix 48-49, 54 Hyalina 34, 48 Retinella (Glyphyalinia) 49 Vitrea 48 indecora, Paradoris 281 inflatus, Potamilus 88 influcata, Quadrula 99 insignis, Luntia 37-38, 46, 57 insularum, Pomacea 19 intermedius, Arion 29-30 isabella, Dentellaria 51 Helicogena 50 Helix 50-51 Helix (Dentellaria) 51 Pleurodonte 34, 36, 38, 50-52, 57 Pleurodonte (Caprinus) 51 Islamia 103, 106, 125, 131, 133-134, 143, 158, 162-163, 165-166, 169-170, 172, 177, 179, 198, 198-201, 201, 203 (Adriolitorea) 165 anatolica 175, 178 ateni 175, 178, 182, 184 azarum 175, 178 bendidis 175, 179 bosniaca 175, 178 burnabasa 175, 178 cianensis 175, 178, 202 consolationis 103, 155, 175, 178, 184, 199-201, 201, 205, 206, 207 coronadoi 175, 179 epirana 175, 179 fezi 175, 179 gaillardoti 175, 179 gaiteri 175, 179, 202 globulina 103, 175-177, 179, 184, 195, 198, 199-203, 201, 204 globulus globulus 175, 179, 183-184 globulus lagari 176, 179, 183 graeca 175, 179 (Islamia) servaini 133-134 latina 176, 179 mienisi 176, 179 minuta 103, 134, 176, 179, 184, 195, 196-198, 198-201, 201, 203, 205, 207 pseudorientalica 176, 179 pusilla 176-177, 179, 201 schuelei 176, 179 servaini 133-134 INDEX 357 spirata 103, 176, 179, 184, 198, 199-201, 201, 203, 204, 205 trichoniana 177, 179 valvataeformis 131, 132, 133-134, 176- 177,179 zermanica 134, 177, 179 Islamiinae 134 islamioides, Sardohoratia 153, 175, 179 Isognomostoma isognomostoma 1-9, 2, 4, 8 isognomostoma, Isognomostoma 1-9, 2, 4, 8 issaeffi, Upogebia 313 Istriana 163-164, 169-170 jadertina, Hauffenia 171, 175, 178 jamaicensis, Orthalicus undatus 44 Oxystyla undatus 44 Pupa 41 Jorunna 256, 294 Jousseaumiella 323 kambeul, Limicolaria 225-226, 232-234 karamani, Lyhnidia 141, 175, 179 karevi, Ohrigocea 147, 175, 179 Ohrigocea (Ohrigocea) 175 Karevia 134-136, 141, 147, 158, 162-163, 165-166, 168, 173 ornata 132, 134-136, 176, 179 kerguelenensis, Archidoris 244 Austrodoris 237, 240, 244, 246-247, 248, 2525255, 308 Cadlina 243 Kerkia 136, 138, 158, 162-163, 165-166, 169-170, 173 brezicensis 138, 175, 179 kusceri 117, 128, 136, 137, 138, 176, 179 kerschneri, Hauffenia 103, 171, 175-176, 178, 180-181, 185, 186 Hauffenia kerschneri 182, 186 Horatia erythropomatia 175, 181 klecakiana, Horatia 127, 128, 130, 130-131, 175-178 knorn, Horatia 178 knorri, Horatia 131, 175 kobeltiana, Parabaikalia florii 60, 68, 80-81 korotnevi, Korotnewia 71, 76, 80, 82 Korotnewia 65 korotnevi 71, 76, 80, 82 semenkewitschi 60, 71, 76, 80-82 krkae, Hadziella 122, 176, 178 kusceri, Hauffenia 136, 176 Horatia 156, 176 Kerkia 117, 128, 136, 137, 138, 176, 179 Zaumia 156-157, 176, 179 labio, Monodonta 224, 226, 232-234 laboutei, Austrodoris 240, 252, 253-254, 255, 309 Lacteoluna (Aerotrochus) turbiniformis 50 selenina 38, 50, 57 selenina barbadensis 50 subaquila 38 turbiniformis 38 lacustre, Musculium 337 lacustris, Pseudamnicola (Rotondia) 176 Pseudohoratia 152, 176, 179 laeve, Deroceras 26 Laevicaulis alte 225, 232-234 laevis, Cadlina 238 lagari, Islamia globulus 176, 179, 183 Neohoratia globulus 103, 177 Pseudamnicola 176 lamellata, Leptinaria 38, 46, 54, 57 Lampsilis 94 cariosa 94 radiata 94 Lanzaiopsis 163-164, 169-170 latina, Islamia 176, 179 Leptinaria 46 antillarum 46 lamellata 38, 46, 54, 57 Leptodea ochracea 94 letourneuxi, Horatia 130-131, 176 leuca, Paradoris 284, 287 lienosa, Villosa 89-90, 92-93 lilljeborgi, Pisidium 337 Limax maximus 25 Limicolaria kambeul 225-226, 232-234 Liobaicalia stiedae 59-60, 62-65, 66, 68, 69, 71, 73, 76-77, 79-83 lippa, Peltodoris 277, 278-280, 281 Lithoglyphulidae 111 Lithoglyphus 163-164, 169-170 parvulus 176 Litigiella glabra 323 Litthabitella 163-164, 169-170 Littoridininae 168 Littorinidae 224 ljovuschkini, Horatia 176, 178 Lobaunia 103, 138-139, 161-163, 165, 168- 170, 180 danubialis 103, 138, 175, 180 loichiana, Hauffenia kerschneri 176, 181- 358 182, 185-186 Lucidella barbadensis 33, 37-38, 40, 53-54 holoserica 40 plicatula 38, 40, 53 rugosa 38 Lucidella (Poeniella) barbadensis 39-40 plicatula 40 lucidula, Hauffenia 171-172, 176, 178 Horatia (Hauffenia) 176 lucidulus, Horatia (Hauffenia) 176 Ludwigia 16 lugubris, Dendrodoris 302 Luntia insignis 37-38, 46, 57 lusitanicus, Arion 25-31, 28-29 lutea, Geitodoris 271 Lyhnidia 139, 141, 154, 158, 162, 164-166, 169-170, 173 gjorgjevici 141, 175, 179 hadzii 139, 140, 141, 175, 179 karamani 141, 175, 179 stankovici 141, 177, 179 sublitoralis 141, 177, 179 lyhnidica, Pseudamnicola (Ohrigocea) 176 Lymnaea auricularia 225, 232-234 glabra 225, 227, 232-234 stagnalis 225, 227, 232-234 viator 16 Lymnaeidae 16, 223-224, 227, 232-234 Maackia 70, 71, 83 bythiniopsis 64-65, 67, 68, 71, 71, 76, 80-83 costata 65, 71, 80-81 (Eubaicalia) bythiniopsis 60 (Eubaicalia) herderiana 59-60 (Eubaicalia) variesculpta 60 herderiana 62-65, 68, 70, 71, 76-83 (Maackia) costata 60 vanesculpta 63-65, 68, 70, 71, 76-77, 80-83 macedonica, Horatia 131, 176, 178 Sadleriana 176 macmurdensis, Austrodoris 248 Macrospira aperta 47-48 macrostoma, Teratobaikalia 64-65, 67, 68, 71, 72, 76, 79-83 Teratobaikalia (Teratobaikalia) 60 magellanica, Cadlina 243 major, Upogebia 313-317, 314-315, 322-323 Malacolimax tenellus 25 malaprespensis, Prespolitorea 150, 176, 179 Mandarina aureola 9 ponderosa 9 INDEX Marstoniopsis 163-164, 169-170 matheroni, Omalonyx 223-225, 232-234 Mathilda 328 amanda 328, 329, 331-334 Mathildidae 327, 329 maximus, Limax 25 media, Hauffenia 103, 171, 176, 178, 186, 187, 192 Megalobulimidae 49 Megalobulimus oblongus 38, 49-50, 57 Megalonaias 94 Megalovalvata 65 Mercuria 163-164, 169-170 michleri, Hauffenia 103, 122, 123, 124-127, 128-129, 176, 189-190 micra, Allopeas 38, 45, 54, 57 Helix (Achatina) 45 Stenogyra 45 Valvata 179 micrometrica, Valvata 172, 176, 202-203 Microna ateni 175 Microphysa vortex 50 mienisi, Islamia 176, 179 Mienisiella 134, 176 Mienisiella 134, 165 gaillardoti 134 mienisi 134, 176 miladinovorum, Ohrigocea 147, 176, 179 Ohrigocea (Karevia) 136, 176 milium, Pisidium 337 miljackae, Dalmatella 176 mimetica, Baptodoris 302 minor, Pineria viequensis 45 minuta, Hauffenia 173, 198 Hauffenia (Neohoratia) 173 Horatia 173 Horatia minuta 173 Islamia 103, 134, 176, 179, 184, 195, 196-198, 198-201, 201, 203, 205, 207 Ohridohauffenia 145, 176, 179 Pseudamnicola 176 Valvata 172, 176, 195, 199-200, 202 minutissima, Caecilianella 47-48 Caecilianella consobrinus 47 Caecilianella (Caecilioides) consobrinus 48 Caecilioides 47 Cecilioides consobrinus 38, 53 Cecilioides (Karolus) consobrinus 47 Miradiscops 49 implicans 38, 49, 53 modicum, Pleurobema 88 Moitessieria 163-164, 169-170 moitessierianum, Pisidium 337-344, 341- 343 Monodonta labio 224, 226, 232-234 Montacuta phascolionis 322 tenella 324 Montacutidae 313, 323-324 Montacutona 324 moquiniana, Válvata 172-173, 176, 202-203 morosa, Diaulula 289, 291 Musculium lacustre 337 transversum 337 Mysella bidentata 323-324 cuneata 323 phascolionis 323 naegelei, Daudebardiella 176, 178 nana, Teratobaikalia 65, 68, 71, 73 Teratobaikalia (Baicaliella) 60 Teratobaikalia nana 73 narutensis, Upogebia 313 naumi, Pseudamnicola (Rotondia) 145 Naumia 106, 145, 165 nemoralis, Cepaea 9 Neohoratia 103-104, 127, 134, 139, 141, 144, 161-162, 164-165, 168-171, 173- 174, 177, 189 ateni 103, 177 azarum 175, 177 coronadoi 177 epirana 171-172 fezi 177 gasulli 177 globulina 198 globulus 177 globulus globulus 103, 177 globulus lagari 103, 177 hadei 171 schuelei 177 subpiscinalis 104, 141, 144, 161, 165, 189 Neopisidium 343 nigra, Dendrodoris 306 nigrobranchiata, Cadlina 242 Nipponomysella subtruncata 323 nitidum, Pisidium 337 Sphaerium 337 noumeae, Peltodoris 281 novoselensis, Horatia 131, 176, 178 nugax, Valvata micra 179 INDEX 359 obliqua, Horatia 130-131, 176 oblongus, Bulimus 49 Bulimus (Borus) 49 Helix 49 Strophocheilus (Borus) 49 Megalobulimus 38, 49-50, 57 obtusa, Horatia 130-131, 176 obtusale, Pisidium 337 occidentalis, Helicina 37 ochracea, Leptodea 94 ochridana, Pseudohoratia 117, 128, 150, 151,152, 176, 179 Valvata 150 Valvata (Atropidina) 150, 176 octona, Achatina 46 Stenogyra 47 Subulina 38, 46-47, 57 octonoides, Bulimus 45 Opeas 45 Stenogyra 45 octonus, Bulimus 46 ohridana, Gocea 116, 118-120, 119, 176, 178 Strugia 153-154, 176, 179 Ohridohauffenia 135, 144-145, 147, 158, 162, 164-166, 168, 173 depressa 144, 175, 179 drimica 145, 175, 179 minuta 145, 176, 179 rotonda 145, 176, 179 sanctinaumi 145, 176, 179 sublitoralis 145, 177, 179 Ohridohoratia 144 gjorgjevici 144 (Ohridohauffenia) 165 (Ohridohauffenia) gjorgjevici 105, 144- 145, 175 Ohridosturanya 106, 147, 165 Ohrigocea 134-135, 145, 147, 158, 162, 164-166, 168, 173 karevi 147, 175, 179 (Karevia) 165 (Karevia) miladinovorum 136, 176 (Karevia) prlitchevi 105, 134-136, 176 (Karevia) sandanskii 135-136, 176 miladinovorum 147, 176, 179 (Ohrigocea) 165 (Ohrigocea) karevi 175 (Ohrigocea) samuili 145, 176 prlitchevi 134, 136 samuili 145, 146, 147, 176, 179 stankovici 147, 176-177, 179 360 ohshimai, Geitodoris 271 Peregrinamor 313-319, 314-315, 318- 320, 321-324 Omalogyridae 328 Omalonyx 224, 227 matheroni 223-225, 232-234 Onagraceae 16 Onchidella celtica 225, 232-234 Onchididae 227, 232 Opeas 38, 45 ascendens 46, 53 beckiana 46 goodalli 46, 53 gundlachi 47 hannense 38, 45-46, 53, 57 octonoides 45 subula 45 orbicularis, Dendrodoris 278, 302, 303, 304, 309 Orbitestellidae 328 Orientalina 163-164, 169-170 ornata, Dolapia 135 Karevia 132, 134-136, 176, 179 Pseudamnicola 105, 134-136, 176 Pseudamnicola (Karevia) 135 Orthalicus 44 maracaibensis subpulchella 38, 43-44, 57 undatus jamaicensis 44 zebra 43-44 oviformis, Parabaikalia 60, 63-65, 66, 68, 71, 73, 74, 76-77, 79-83 Oxyloma 225, 227, 232-234 Oxystyla 44 maracaibensis subpulchella 44 pulchella 44 undatus jamaicensis 44 Palacanthiliopsis 163-164, 168-170 Paladilhia 163-164, 169-170 Paladilhiopsis 163-164 pallida, Geitodoris 240, 260, 268-269, 269-270, 271 palma, Discodoris 271 Paludina sturmi 174, 177 paludosa, Pomacea 19 palustris, Horatia 130-131, 176 Stagnicola 225, 227, 232-234 pangaea, Daphniola exigua 176, 178 Horatia (Daphniola) exigua 113-114, 176 Parabaikalia 74, 81 elata 63, 79 INDEX elata dubiosa 60, 71, 76, 80-82 elata elata 60, 64-65, 68, 71, 76, 79-82 florii 62-65, 68, 71, 74, 79 florii florii 60, 64-65, 68, 71, 76, 79-82 florii kobeltiana 60, 68, 80-81 oviformis 60, 63-65, 66, 68, 71, 73, 74, 76-77, 79-83 Paradoris 237, 256, 281, 284, 307-308 araneosa 257, 281, 282-283, 284-285, 287-288, 309 granulata 281 imperfecta 257, 285, 286-287, 287-288 indecora 281 leuca 284, 287 tsurugensis 284, 287 parchappii, Chilina 16 Heleobia 16 parvula, Horatia 176, 178 parvulus, Lithoglyphus 176 Pauluccinella 163-164, 169-170 peggyae, Utterbackia 99 pellucida, Gastrocopta 38, 42 Pupa 41 Peltodoris 237, 277, 281, 307 atromaculata 277, 281 fellowski 281, 284 lippa 277, 278-280, 281 noumeae 281 peregra, Radix 225, 232-234 peregrina, Biomphalaria 16 Peregrinamor ohshimai 313-319, 314-315, 318-320, 321-324 perplexa, Helix 51 Helix (Dentellaria) 51 Helix (Helicodonta) 51 Pleurodonte 51 Polydontes 38, 51 pescei, Arganiella 106-108, 107, 110, 176, 178 Pezzolia 146, 147, 149, 157, 158, 161-162, 164-166, 169-170, 173 radapalladis 128, 146, 147-149, 176, 179 Pharodoris 237, 256-257, 260, 263, 266- 268, 267, 307 diaphora 240, 257, 260, 261-263, 264, 266, 307, 309 philippinensis 240, 263, 264, 265-266, 266-267 phascolionis, Montacuta 322 Mysella 323 Phasmarhabditis hermaphrodita 30 philippinensis, Pharodoris 240, 263, 264, 265-266, 266-267 phinei, Baptodoris 278, 299, 300-301, 302, 308-309 Pholadidae 323 Pholadoidea 323 Phreatica 163-164, 169-170 Phreatodrobia 180 Phyllidia 237 Phyllidiidae 237 Phyllidiopsis 237, 308 Physa 16 Physidae 16 Pineria viequensis 45 viequensis minor 45 piscinalis, Valvata 199 Pisidium 337-338, 343, 346 amnicum 337 casertanum 337 compressum 340 cruciatum 340 henslowanum 337 lilljeborgi 337 lilljeborgi cristatum 340 milium 337 moitessierianum 337-344, 341-343 nitidum 337 obtusale 337 punctatum 337-340, 339, 343-345, 344 punctiferum 337-340, 343-344, 346, 346 simplex 337-340, 342-344, 345-346 subtruncatum 337 supinum 337 tenuilineatum 337 Plagigeyeria 163-164, 169-170 plana, Hauffenia 171, 176, 178 Planorbidae 65, 223, 227, 232-234 planospira, Fissuria 116, 176, 178 Platydoris galbanus 284 Plekocheilus aurissileni 43 Plekocheilus (Plekocheilus) aurissileni 43 Pleurobema 87-90, 91, 92-94, 94, 95, 97- 98, 97-99 bulbosum 87-90, 91, 92-94, 94, 97, 97- 98, 99 clava 87-90, 92, 96, 98 modicum 88 pyriforme 87-90, 91, 92-94, 94, 96, 97- 99, 97-98 reclusum 87-90, 91, 92-94, 94, 97-99, 97-98 sintoxia 87, 89-90, 92, 96, 97-98 INDEX 361 strodeanum 87-90, 91, 92-93, 94, 96, 97-98 Pleurodonte (Caprinus) isabella 51 dentiens 38, 51 isabella 34, 36, 38, 50-52, 57 perplexa 51 plicatula, Helicina 40 Lucidella 38, 40, 53 Lucidella (Poeniella) 40 polinskii, Horatia 152, 176 Polydontes perplexa 38, 51 polymorpha, Dreissena 87, 337 Pomacea 15, 19, 22 canaliculata 13-22, 18-19, 21 insularum 19 paludosa 19 Potamilus inflatus 88 Potamopyrginae 168 Potamopyrgus 163-164, 168-170 Potidoma subtrigonum 323 praeclara, Horatia 130-131, 176 Prespolitorea 106, 149-150, 158, 162, 164- 166, 168, 173 malaprespensis 150, 176, 179 valvataeformis 146, 149-150, 177, 179 prlitchevi, Ohrigocea 134, 136 Ohrigocea (Karevia) 105, 134-136, 176 producta, Teratobaikalia nana 68, 73 Teratobaikalia (Baicaliella) nana 60 provisoria, Helix 51-52 Zachrysia 37-38, 51-52, 54, 57 Pseudamnicola 163-164, 169-170 depressa 105, 144, 175 (Karevia) ornata 135 minuta 176 (Ohridohauffenia) sublitoralis 177 (Ohrigocea) Iyhnidica 176 ornata 105, 134-136, 176 rotonda 176 (Rotondia) 165 (Rotondia) drimica 175 (Rotondia) lacustris 176 (Rotondia) naumi 145 (Rotondia) rotonda 145 (Rotondia) st. naumi 176 Pseudavenionia 163-164, 169-170 Pseudobaikalia contabulata 65, 76, 80, 82 (Microbaicalia) pulla pulla 60 (Microbaicalia) pulla tenuicosta 60 (Pseudobaikalia) contabulata 60 (Pseudobaikalia) zachwatkini 60, 63 pulla 71 362 pulla pulla 76, 82 pulla tenuicosta 80 zachwatkini 63, 65, 66, 68, 69, 71, 76, 79-82 Pseudohoratia 150, 152, 157, 158, 162, 164-166, 169-170, 173 brusinae 152, 175, 179 lacustris 152, 176, 179 ochridana 117, 128, 150, 151, 152, 179 Pseudoislamia 152, 158, 162, 164-166, 168, 173 balcanica 152, 175, 179 Pseudopineria barbadensis 33, 37-38, 45, 53-54 pseudorientalica, Islamia 176, 179 pulchella, Achatina 44 Godlewskia 60, 64-65, 68, 71, 76, 79-82 Oxystyla 44 pulla, Pseudobaikalia 71 pulla Pseudobaikalia 76, 82 pulla Pseudobaikalia (Microbaicalia) 60 pumilus, Bulimus 46 punctatum, Pisidium 337-340, 339, 343- 345, 344 punctiferum, Pisidium 337-340, 343-344, 346, 346 Pupa barbadensis 41-42 bicolor 48 jamaicensis 41 pellucida 41 Pupisoma dioscoricola 53 (Ptychopatula) dioscoricola 42 pusae, Geitodoris 271 pusilla, Islamia 176-177, 179, 201 Upogebia 323 Valvata 176 putris, Succinea 223 Pyramidellidae 328 Pyramidelloidea 327 pyriforme, Pleurobema 87-90, 91, 92-94, 94, 96, 97-98, 97-99 Quadrula 89 apiculata 89-90, 98 influcata 99 Radix peregra 225, 232-234 raehlei, Fissuria 155, 172, 176, 178, 193, 194 Hauffenia 171-172 Horatia (Hauffenia) 103, 116, 176, 193, 195 INDEX radapalladis, Pezzolia 128, 146, 147-149, 176, 179 radiata, Lampsilis 94 reclusum, Pleurobema 87-90, 91, 92-94, 94, 97-98, 97-99 repanda, Doris 238 reticulatum, Deroceras 25-26, 31 Retinella (Glyphyalinia) carolinensis 49 (Glyphyalinia) incisa 49 Rostanga 237, 294-296, 307-308 ankyra 278, 294, 295-296, 296 rubra 294 setidens 295-296 rotonda, Ohridohauffenia 145, 176, 179 Pseudamnicola 176 Pseudamnicola (Rotondia) 145 Rotondia 106, 145, 165 rubescens, Archidoris 244 rubra, Diaulula 289 Rostanga 294 rudnicae, Haaziella 122, 176, 178 rugosa, Helicina 40 Lucidella 38 Sacculina sacculinae 314 upogebiae 314 sacculinae, Sacculina 314 Sacharomyces cerevisiae 221 Sadleriana 163-164, 169-170 macedonica 176 Sagdidae 50 salsa, Spurwinkia 138 samuili, Ohrigocea 145, 146, 147, 176, 179 Ohrigocea (Ohrigocea) 145, 176 sanctinaumi, Ohridohauffenia 145, 176, 179 sanctizaumi, Zaumia 157, 177, 179 sandanskii, Ohngocea (Karevia) 135-136, 176 sandiegensis, Diaulula 291, 294 Doris 288-289 Sardohoratia 152-153, 158, 162, 164-166, 169-170, 174 islamioides 153, 175, 179 sulcata 152-153, 177, 179 Sardopaladilhia 163-164, 169-170 Helix 50 Lacteoluna 38, 50, 57 schuelei, Hauffenia (Neohoratia) coronadoi 176 Islamia 176, 179 Neohoratia 177 Sclerodoris 237, 256, 296, 299, 307-308 tuberculata 296 virgulata 278, 296, 297-298, 299, 309 semenkewitschi, Korotnewia 60, 71, 76, 80- 82 semidecussatus, Tapes 217-218, 220 servaini, Horatia 131, 133-134, 176 Islamia 133-134 Islamia (Islamia) 133-134 setidens, Rostanga 295-296 Boreodoris 295 Sheitanok 165 amidicus 175, 179 similaris, Bradybaena 38, 52, 57 Eulota 52 Fruticicola 52 Helix 52 Helix (Dorcasia) 52 Hélice (Hélicelle) 52 Helix (Fruticicola) 52 similis, Solatopupa 9 simplex, Pisidium 337-340, 342-344, 345- 346 sinjana, Hauffenia 171, 176, 178 Hauffenia jadertina 176 sintoxia, Pleurobema 87, 89-90, 92, 96, 97-98 Siphonaria 227 algesirae 225, 232-234 Siphonariidae 227, 232 Siphonostoma costata 44 sketi, Dalmatella 111, 176, 178 Haaziella 122, 176, 178 sokolovi, Horatia 176, 178 Solatopupa similis 9 solitaria, Hauffenia 171, 176 sparsa, Cadlina 243 Sphaeriidae 337 Sphaerium corneum 337 nitidum 337 Sphonariidae 234 spirata, Hauffenia minuta 176, 203 Horatia minuta 173 Islamia 103, 176, 179, 184, 198, 199- 201, 201, 203, 204, 205 spiridoni, Bracenica 108-109, 176, 178 Spurwinkia salsa 138 st. naumi, Pseudamnicola (Rotondia) 176 st. zaumi, Horatia 177 stagnalis, Lymnaea 225, 227, 232-234 Stagnicola palustris 225, 227, 232-234 stankovici, Horatia (Hauffenia) 147, 177 Lyhnidia 141, 177, 179 Ohrigocea 147, 176-177, 179 INDEX 363 Stenogyra beckiana 45 caraccasensis 45 goodalli 46 gundlachi 47 micra 45 octona 47 octonoides 45 subula 45 viequensis 45 sticta, Geitodoris 271 stiedae, Liobaicalia 59-60, 62-65, 66, 68, 69, 71, 73, 76-77, 79-83 Streptaxidae 48 Streptaxis deformis 48 glaber 34, 38, 54, 57 glabra 48 (Odontertemon) glaber 48 (Streptartemon) glaber 48 strodeanum, Pleurobema 87-90, 91, 92-93, 94, 96, 97-98 Strophocheilus (Borus) oblongus 49 Strugia 106, 153-154, 158, 162, 164-166, 168, 173 ohridana 153-154, 176, 179 sturmi, Horatia 174, 177-178 Paludina 174, 177 subaquila, Helix 50 Lacteoluna 38 subcarinata, Hauffenia 103, 128, 171, 177- 178, 180, 188, 191 Hauffenia (Hauffenia) 177, 188 subfuscus, Arion 25, 30 sublitoralis, Lyhnidia 141, 177, 179 Ohridohauffenia 145, 177, 179 Pseudamnicola (Ohridohauffenia) 177 subpiscinalis, Hauffenia 103-104, 115, 117- 118, 128-129, 140, 142, 144, 161, 171, 177-178, 180, 189 Neohoratia 104, 141, 144, 161, 165, 189 Valvata 103-104, 134, 141, 144, 177, 189 subpulchella, Orthalicus maracaibensis 38, 43-44, 57 Oxystyla maracaibensis 44 substriata, Helicina 37, 39 Helicina fasciata 36-39, 57 subtrigonum, Potidoma 323 subtruncata, Nipponomysella 323 subtruncatum, Pisidium 337 subula, Bulimus 45 Opeas 45 Stenogyra 45 364 Subulina 47 octona 38, 46-47, 57 Subulinidae 45 Succinea 41, 224, 233 barbadensis 38, 41, 57 bermudensis 38, 41 putris 223 Succineidae 41, 223-224, 227, 232-233 sulcata, Sardohoratia 152-153, 177, 179 supinum, Pisidium 337 supracarinata, Horatia 177, 188 symmetros, Axinodon 324 Systrophiidae 49 Tapes semidecussatus 217-218, 220 Taringa 299 tatei, Brachypodella 45 tellinii, Hauffenia 103, 110, 117-118, 122, 123-124, 127, 128-129, 161, 165, 171, 176-178, 180, 189-190, 192 Horatia 103-104, 144 Horatia (Hauffenia) 103, 122, 177, 189-190 tenella, Montacuta 324 tenellus, Malacolimax 25 tenuicosta, Pseudobaikalia pulla 80 Pseudobaikalia (Microbaicalia) pulla 60 tenuilineatum, Pisidium 337 tenuissimus, Bulimulus 38, 43 Teratobaikalia 82 (Baicaliella) nana 60 (Baicaliella) nana clandestina 60 (Baicaliella) nana humerosa 60 (Baicaliella) nana producta 60 ciliata 64-65, 67, 68, 71, 72, 76, 79-83 duthiersii 65, 68, 71, 72, 76, 80-83 macrostoma 64-65, 67, 68, 71, 72, 76, 79-83 nana 65, 68, 71, 73 nana clandestina 68, 73, 76, 79-80, 82 nana humerosa 68, 73, 82 nana nana 73 nana producta 68, 73 (Teratobaikalia) macrostoma 60 (Trichiobaikalia) 72 (Trichiobaikalia) ciliata 60 (Trichiobaikalia) duthiersii 60 Tetraselmis tetrathele 315 tetrathele, Tetraselmis 315 Thais clavigera 224-226, 232-234 thermalis, Hadziella 122, 177-178 Thyasiridae 324 INDEX Thysanophora 50 turbiniformis 50 vortex 50 Tornatellina antillarum 46 tovunica, Hauffenia 103, 161, 171, 177- 178, 180, 190, 191, 192 Hauffenia (Hauffenia) 177 transversum, Musculium 337 Trichiobaikalia 82 trichoniana, Islamia 177, 179 Trochidae 328 troglobja, Valvata 177 Truncatella 38, 40-41 barbadensis 38, 40, 57 bilabiata 40 (Truncatella) bilabiata 40 Truncatellidae 40 truncatula, Fossaria 225, 232-234 tsurugensis, Paradoris 284, 287 tuberculata, Sclerodoris 296 turbiniformis, Helix 50 Helix (Microphysa) 50 Lacteoluna 38 Lacteoluna (Aerotrochus) 50 Thysanophora 50 Turbo uva 42 turgidula, Valvata 172, 177, 202-203 turriformis, Baicalia 60, 65, 66, 68, 75, 81 umbilicata, Haaziella 177 Umbraculum umbraculum 307 umbraculum, Umbraculum 307 Uniomerus 89, 94, 94, 97 caroliniana 90, 92-93, 96, 97 Unionidae 87 Upogebia 313-316, 318, 322 issaeffi 313 major 313-317, 314-315, 322-323 narutensis 313 pusilla 323 upogebiae, Sacculina 314 Urocoptidae 44 Urocoptis 44 Utterbackia peggyae 99 uva, Cerion 38, 42 Turbo 42 Valvata 103, 131, 172 (Atropidina) ochridana 150, 176 bourguignati 172, 175, 202-203 (Cincinna) gaillardoti 175 (Cincinna) hellenica 111, 113, 175 coronadoi 175 erythropomatia 103-104, 114-115, 175, 180 exigua 111, 113, 175 exilis 108, 172-175, 200, 208-209 globulina 172-173, 175, 199-200, 202-203 micra 179 micra nugax 179 micrometrica 172, 176, 202-203 minuta 172, 176, 195, 199-200, 202 moquiniana 172-173, 176, 202-203 ochridana 150 piscinalis 199 pusilla 176 subpiscinalis 103-104, 134, 141, 144, 177,189 troglobja 177 (Tropidina) fezi 175 turgidula 172, 177, 202-203 wagner 103-104, 154, 156, 177, 192 ılvataeformis, Horatia 103, 126 Horatia (Hauffenia) 122, 177, 189-190 Hydrobia 131, 133-134, 177 Islamia 131, 132, 133-134, 176-177, 179 Prespolitorea 146, 149-150, 177, 179 alvatidae 65, 328 alvatoidea 327 ariesculpta, Maackia (Eubaicalia) 60 Maackia 63-65, 68, 70, 71, 76-77, 80-83 ardica, Erythropomatiana 115, 140, 141, 143-144, 161, 177, 189 Hauffenia 189 arlikana, Horatia 130-131, 177 ertiginidae 41 2stita, Doris 291 ator, Lymnaea 16 equensis, Bulimus 45 Pineria 45 Stenogyra 45 Поза 89, 94 lienosa 89-90, 92-93 INDEX 365 virgulata, Sclerodoris 278, 296, 297-298, 299, 309 Vitrea 49 implicans 49 incisa 48 Voluta auris 43 vortex, Helix 50 Helix (Microphysa) 50 Microphysa 50 Thysanophora 50 Vrania 103-104, 127, 139, 154, 156, 161- 162, 164-165, 168-171 wagneri 154, 161, 165, 192 wagneri, Hauffenia 103, 117, 128, 151, 155, 171, 177-178, 180, 186, 188, 192 Valvata 103-104, 154, 156, 177, 192 Vrania 154, 161, 165, 192 wienerwaldensis, Hauffenia 103, 171, 177- 178, 180, 191, 192 willani, Cadlina 242 Xylophaga dorsalis 323 Zachrysia provisoria 37-38, 51-52, 54, 57 zachwatkini, Pseudobaikalia 63, 65, 66, 68, 69, 71, 76, 79-82 Pseudobaikalia (Pseudobaikalia) 60 Zaumia 106, 156-157, 158, 162, 164-166, 168, 173 kusceri 156-157, 176, 179 sanctizaumi 157, 177, 179 zebra, Buccinum 44 Orthalicus 43-44 zermanica, Islamia 134, 177, 179 Zonites decolorata 49 implicans 49 Zonitidae 48 MALACOLOGIA International Journal of Malacology Vol. 43(1-2) 2001 Vol. Vol. Vol Vol. Vol. Vol. Vol. Vol. Vol. Vol. Vol. Vol. Vol. Publication dates 33, No. 34, No. ‚ 35, №. 35, Мо. 36, No. S7.No: 37, No. 38, No. 39, No. 40, No. 41, No. 41, No. 42, No. то 122 6 Sep. 1991 9 Sep. 1992 14 Jul. 1993 2 Dec. 1993 8 Jan. 1995 13 Nov. 1995 8 Mar. 1996 17 Dec. 1996 13 May 1998 17 Dec. 1998 22 Sep. 1999 31 Dec. 1999 18 Oct. 2000 VOL. 43, NO. 1-2 MALACOLOGIA 2001 CONTENTS MARCO BODON, GIUSEPPE MANGANELLI & FOLCO GIUSTI A Survey of the European Valvatiform Hydrobiid Genera, with Special Reference to Hauffenia Pollonera, 1898 (Gastropoda: Hydrobiidae) ...... 103 RONALD CHASE & DAVID G. ROBINSON The Uncertain History of Land Snails on Barbados: Implications for CONSERVATION a ere ree ee И 33 ROBERT CIPRIANI & RUDIGER BIELER Eyeballs and Pitfalls: Estimating the Degree of Heterostrophy in the Heterobranch Shell (Gastropoda: Heterobranchia) .................... 327 ANA VIRGINIA CALOGERAS DUTRA-CLARKE, CHARLENE WILLIAMS, REBECCA DICKSTEIN, NORBERT KAUFER & JAMES R. SPOTILA Inferences on the Phylogenetic Relationships of Succineidae (Mollusca, Pulmonata) Basedion 18$ rRNA Gene... 0.002... 223 BRIGITTE GRIMM Life Cycle and Population Density of the Pest Slug Arion lusitanicus Mabille (Mellusca#Pulmonata)ion/Grassland oia 6.28 ee 25 KAREN L. KANDL, HSIU-PING LIU, ROBERT S. BUTLER, WALTER R. HOEH, & MARGARET MULVEY A Genetic Approach to Resolving Taxonomic Ambiguity among Pleurobema (Bivalvia: Unionidae) of the Eastern Gulf Coast ...................... 87 ALEXEI V. KORNIUSHIN, IGOR A. GRIGOROVICH & GERALD L. MACKIE Taxonomic Revision of Pisidium punctatum Sterki, 1895 (Bivalvia: SAVIA AG wy ewes A OU OR Е 337. JORGEN LUTZEN, HIROSHI SAKAMOTO, AYAKO TAGUCHI & TOHRU TAKAHASHI Reproduction, Dwarf Males, Sperm Dimorphism, and Life Cycle in the Commensal Bivalve Peregrinamor ohshimai Shöji (Heterodonta: Galeommatoidea:Montacutidae): еее cee ute se se 313 PABLO R. MARTÍN, ALEJANDRA L. ESTEBENET & NESTOR J. CAZZANIGA Factors Affecting the Distribution of Pomacea canaliculata (Gastropoda: Ampullariidae) along Its Southernmost Natural Limit .................. 13 L. MERCADO, E. ITARTE, S. MARSHALL & G. ARENAS Immunological Detection of a CK2 Activity in Tapes semidecussatus (Molitisca Bivalvia) Майе... 2% ohe sper о аи 217. T. SITNIKOVA, P. RÖPSTORF & F. RIEDEL Reproduction, Duration of Embryogenesis, Egg Capsules and Protoconchs of Gastropods of the Family Baicaliidae (Caenogastropoda) Endemic to KakerBaikalb es va о о 59 ANGEL VALDES Deep-Sea Cryptobranch Dorid Nudibranchs (Mollusca, Opisthobranchia) from the Tropical West Pacific, with Descriptions of Two New Genera and EIGinteeniINGW: SPECIES. a5 Su ke ee ee 237 P. VAN RIEL, К. JORDAENS, J. Е. VAN GOETHEM & T. BACKELJAU Genetic Variation in the Land Snail /sognomostoma isognomostoma (Gastropoda:¡Pulmonatas Helicidae) cum rio een 1 WHY NOT SUBSCRIBE TO MALACOLOGIA ORDER FORM Your name and address Send U.S. $56.00 for a personal subscription (per volume) or U.S. $70.00 for an institutional subscription. VISA and MASTERCARD are accepted for an addi- tional $2.00 fee. Checks must be drawn on an American bank and made payable to MALACOLOGIA. Address: Malacologia P.O. Box 385 Haddonfield, NJ 08033-0309 U.S.A. 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There is a $4.50 handling charge per volume for all pur- chases of single volumes. Address inquiries to the Subscription Office. VOL. 43, NO. 1-2 MALACOLOGIA CONTENTS P. VAN RIEL, K. JORDAENS, J. L. VAN GOETHEM 4 T. BACKELJAU Genetic Variation in the Land Snail /sognomostoma isognomostoma (Gastropoda: Pulmonata: Helleidae) .. 2. o... <<<: weenie ta 00000 oe PABLO R. MARTIN, ALEJANDRA L. ESTEBENET & NESTOR J. CAZZANIGA Factors Affecting the Distribution of Pomacea canaliculata (Gastropoda: Ampullariidae) along Its Southernmost Natural Limit .................. BRIGITTE GRIMM Life Cycle and Population Density of the Pest Slug Arion lusitanicus Mabille (Mollusca: Pulmonata) on Grassland ....................:6....0 RONALD CHASE & DAVID G. ROBINSON The Uncertain History of Land Snails on Barbados: Implications for ЕВ D nice cb oe ee aa sale ee T. SITNIKOVA, P. ROPSTORF & F. RIEDEL Reproduction, Duration of Embryogenesis, Egg Capsules and Protoconchs of Gastropods of the Family Baicaliidae (Caenogastropoda) Endemic to babe Saka! о. KAREN L. KANDL, HSIU-PING LIU, ROBERT S. BUTLER, WALTER R. HOEH, & MARGARET MULVEY A Genetic Approach to Resolving Taxonomic Ambiguity among Pleurobema (Bivalvia: Unionidae) of the Eastern Gulf Coast ...................... MARCO BODON, GIUSEPPE MANGANELLI 8 FOLCO GIUSTI A Survey of the European Valvatiform Hydrobiid Genera, with Special Reference to Hauffenia Pollonera, 1898 (Gastropoda: Hydrobiidae) ...... L. MERCADO, E. ITARTE, S. MARSHALL & G. ARENAS Immunological Detection of a CK2 Activity in Tapes semidecussatus (Mollusca, Bivalvia) Mme. Las ours oe tad is a er ANA VIRGINIA CALOGERAS DUTRA-CLARKE, CHARLENE WILLIAMS, REBECCA DICKSTEIN, NORBERT KAUFER 8 JAMES В. SPOTILA Inferences on the Phylogenetic Relationships of Succineidae (Mollusca, Pulmonata) Based on 18S: IRNA GONE : :....4uuue conos etes ÁNGEL VALDÉS Deep-Sea Cryptobranch Dorid Nudibranchs (Mollusca, Opisthobranchia) from the Tropical West Pacific, with Descriptions of Two New Genera and Eighlsen = ся dike oem area JORGEN LUTZEN, HIROSHI SAKAMOTO, AYAKO TAGUCHI & TOHRU TAKAHASHI Reproduction, Dwarf Males, Sperm Dimorphism, and Life Cycle in the Commensal Bivalve Peregrinamor ohshimai Shoji (Heterodonta: Galeommataidea: Montaculidas) ...: ок sus aug on Зе ROBERT CIPRIANI & RUDIGER BIELER Eyeballs and Pitfalls: Estimating the Degree of Heterostrophy in the Heterobranch Shell (Gastropoda: Heterobranchia) .................... ALEXEI V. KORNIUSHIN, IGOR А. GRIGOROVICH 8 GERALD |. MACKIE Taxonomic Revision of Pisidium punctatum Sterki, 1895 (Bivalvia: o ae a ae ee aaa sano oo teeta TU 2001 25 33 59 87 103 217 223 237 313 ’ E 2! 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