Chilina megastoma Hylton Scott, 1958 (Pulmonata: Basommatophora) a study on topotypic specimens Cristian F. Ituarte DepartmentofInvertebrateZoology,MuseodeLaPlata, 1900LaPlata,BuenosAires,Argentina,[email protected] Abstract: This isacontribution totheknowledge ofChilinamegastoma Hylton Scott, 1958 (Pulmonata: Basommatophora: Chilinidae), basedon the studyoftopotypicspecimens.ThisspeciesisendemictotheIguazuFalls,afrontierregionbetweenArgentinaandBrazil.C.megastomaisknownonlyfrom abrieforiginaldescription,whichwasbasedexclusivelyontheshellmorphologyofasinglespecimen.Therelevantresultsincludethefollowing: theradula isformedby40-50transverserowsofteethin"chevron"arrangement;thecentraltoothisbicuspidwithareducedorrudimentarythirdcusp; thefirstlateral istricuspidwithawideandshortbase,andtheremaininglateralsandmarginals(40-44)eachhavefourcuspsandalongattachmentbase, which isbent in the marginals. The secondary oraccessory seminal receptacle, in the female genital system, is strongly reduced and does not seems to be functional. The penisisaslongasthepenissheath,andthepenissheathistwoorthreetimeslongerthantheprepuce. Key words: Chilina,morphology,topotypes,endemicspecies,Chilinidae The family Chilinidae comprises only one genus, from the type locality (Castellanos and Landoni, 1995). The Chilina Gray, 1828, which is distributed exclusively in original description was based on the shell morphology ofa South America (Castellanos and Gaillard, 1981; single specimen, thus, details on soft-part anatomy, espe- Castellanos and Miquel, 1991). About 17 Chilina species cially the nervous system, genital system, and radular fea- have been recognized as distributed in Argentina tures, are completely unknown. (Castellanos and Gaillard, 1981; Castellanos and Landoni, In the present paper, the results of the study on 1995). topotypic specimens ofChilina megastoma, focusing on its Previous knowledge on the soft-part anatomy of shell morphometry, radular ultrastructure, anatomy of the members of the genus mainly comes from the studies of male and female genital systems, and nervous system, are Haeckel (1911) and Duncan (1960a, b; 1975). Recently, given. In addition, C. fluminea (Maton, 1809), a species Valdovinos and Stuardo (1995), based upon study of the widely distributed in Argentina, was used for comparative radula, soft-part anatomy, and shell morphology of the 36 analysis ofseveral anatomical features. species ofthe genus described from Chile, proposed a new genus, Archaeochilina, to include Chilina angusta (Philippi, 1860), and recommended the rearrangement of MATERIALS AND METHODS the remaining Chilean species in three new subgenera: Chilina s. s.,Mesochilina, andNeochilina. The authorspro- Two samples of Chilina megastoma were obtained, posed the nervous system organization as providing the one from the vertical cliff behind the Salto Dos Hermanas only reliable character to afford a basis for supraspecific waterfall on the Argentine side of the Iguazu Falls, arrangement. Misiones, Argentina, and the other from a vertical cliffper- Argentine species of Chilina (16 species with three manently swept by a small waterfall at the Brazilian side subspecies) have been studied by Hylton Scott (1958), (the Iguazu River is in the northeastern part of Argentina, Castellanos and Gaillard (1981), Castellanos and Miquel and it constitutes the natural geographical border with (1980), and Miquel (1984; 1986a, b). Brazil at that site). Two other samples were collected from Chilina megastoma Hylton Scott, 1958, was Salto San Martin at the Iguazu Falls and from a small described from the Iguazu Falls (25°35' S, 54°35' W), waterfall at the ArrecheaRivulet, the latter2 km away from Misiones Province, Argentina. The species is known only the type locality. AmericanMalacological Bulletin,Vol. 14(1)(1997):9-15 9 10 AMER. MALAC. BULL. 14(1) (1997) The specimens were fixed in 10% formalin after ratio of last whorl to shell length ranged from 0.92-0.97 having been partly relaxed by immersion in warm water (mean 0.95, standard deviation 0.01). The aperture is oval; (55°C) for a few minutes. After relaxation and fixation in minor diameter to major diameter ratio ranged from 0.53- formalin, the specimens used for anatomical studies were 0.87 (mean 0.67, standard deviation 0.08). The shell is yel- treated and preserved in a modified Raillet-Henry's solu- lowish, olivaceous, or in some cases dark brown. The sur- tion (distilled water 93%, acetic acid 2%, formalin 5%). face is smooth, only sulcated with weak radial periostracal The radulae were dissected from the buccal mass, then striae and helicoidal stripes, particularly evident near the treated with 10% sodium hydroxide (12 hrs), rinsed in dis- suture, which isdeeply marked (Fig.2). Thecolumellarbor- tilled water and properly mounted and coated with gold for der is straight or slightly curved, somewhatconcave, with a scanning electronic microscopic observation. marked fold at its upperend (usually referred to as the col- The type specimen of Chilina megastoma, deposit- umellar tooth). The parietal area has a thin white callus. A ed at the Museo de la Plata (MLP, without registration weak fold of the parietal area forms an indistinct lamella, number), has also been studied. Specimens of C.fluminea alsoreferred to in the literature asa"parietal tooth" (Hylton used for comparative purposes were collected from the Scott, 1958; Castellanos and Gaillard, 1981), lying slightly sandy shore at the Rio de LaPlata Riverand from Miguehn over the columellar fold (Fig. 3). The outer lip is sharp and Rivulet, both at PuntaLara, Ensenada, Buenos Aires. evenly curved. Shell measurements (total shell length, last whorl length, major and minor diameter of the shell aperture) DIGESTIVE SYSTEM: The general morphology were taken under a stereoscopic microscope provided with of the digestive system follows the pattern already known a micrometereyepiece. in the genus (Haeckel, 1911; Harry, 1964). The posterior Voucher specimens were deposited at the malaco- end ofthe radular sac exceeds the posteriorend ofthe buc- logical collection at Museo de La Plata (MLP nos. 5098, cal bulb, forming a short tube slightly bent dorsally. The 5099, 5128, 5129, 5130, 5246, and 5261). salivary glands are not fused attheirposteriorend. The radula is composed of approximately 40 sharply oblique rows of teeth (Fig. 4), that is, the tooth RESULTS rows are arranged "in chevron." The angle between the right and left halves ofeach row is about 90°. The central SHELL: The diagnostic features ofChilina megas- tooth is asymmetrical, bicuspid, with an additional third toma are shape and development of the spire (extremely cusp, greatly reduced, and partly overlapping the major low) and the great development of the last whorl of the cusp (Fig. 5). The attachment base of the central tooth is shell, resulting in a large aperture ofthe shell (Fig. 1). The elongated, deltoid in shape, with the proximal end concave. Figs. 1-3.ShellofChilinamegastomaHyltonScott, 1958.Fig. 1.Aperturalview(MLP5098).Fig.2.Dorsalview,showingtheperiostracalsculpture(MLP 5098).Fig. 3. Detailofthecolumellararea,columellarfold,andparietallamella(MLP5099).Scalebars=2mm. ITUARTE: CHILINA MEGASTOMA 11 In the last marginal teeth, the attachment base becomes slender, obliquely inserted, and slightly bent (Figs. 7-11). REPRODUCTIVE SYSTEM: Regarding the gen- eral aspects, the features ofthe female and male genital sys- tems are coincident with those previously described for the genus (Haeckel, 1911; Duncan, 1960a, b, 1975). Thus, only the diagnostic characteristics for Chilina megastoma will be described. Female genital system: The seminal receptacle or spermatheca is oval, located at the left side of the visceral mass, lyingjustbelow the ventricle. A long duct runs trans- versely across the visceral mass, passing over the utero- vaginal complex, to connect the seminal receptacle with the distal end ofthe vaginajust before the female genital open- ing at the right side ofthe body (Figs. 12-13). Another duct arises posteriorly, just between the vagina and the uterus, Figs. 4-11. Radula ofChilina megastoma Hylton Scott, 1958. Fig. 4. General view showing the typical arrangement of teeth. Fig. 5. Central tooth and first lateral teeth. Figs. 6, 8, 10. Lateral teeth (from central toward marginals). Figs. 1,9, 11. Details for Figs. 6, 8, 10, respectively. Scalebars- 10pm(Fig.5),25pm(Figs.7,9, 11),50pm(Figs.6,8, 10), 250pm(Fig.4). The major cusp is relatively short, cylindrical, and pointed at the tip. The minor cusps are very short and thorn-like. The first lateral tooth is tricuspid, with a short, wide, and strong base. The mesocone has a short dagger-like shape, somewhat directed toward the central tooth. The entocone and ectocone are short, triangular, and nearly symmetrical. The entocone is somewhatlargerthan the ectocone (Figs. 5- 6). The second lateral tooth is also tricuspid, but with a slender, longer, and slightly asymmetric attachment base. Fig. 12-13. Chilinamegastoma Hylton Scott, 1958. Fig. 12. Lateral view The mesocone is the largestcusp, and the ectocone marked- ofapartiallydissectedspecimenshowingtheterminal portionofmaleand ly smaller than the entocone (Fig. 6). From the third lateral femalegenitalsystems,andthedeferentduct(arrowheads)dissectedfrom tooth to the marginal teeth, the morphology of the teeth is thebody wall (MLP5210). Fig. 13. Detailsoftheterminal femalegenital relatively similar. The free plate ofthe teeth is palmate with sityssdtiesmtaslheonwdin(gMtLhePs5p2e1r0m)a.th(eecga,lcdeurcetbrcarlosgsainngglioovne;rtdhde,vdaegfienraenttodoupcetn;aft, four cusps; the attachment base is slender, and it joins the foot; fp, female pore; mp, male pore; p, penis; sd, spermathecal duct; sg, free plate at a point slightly shifted toward the minorcusps. salivarygland;v,vagina). 12 AMER. MALAC. BULL. 14(1) (1997) called a secondary or accessory seminal receptacle. This running below the uterus and the vagina. Near the distal duct runs closely adhered to the wall ofthe uterus, and it is end of the vagina the deferent duct emerges from the barely enlarged at the tip toform a nearly indistinguishable, haemocoelic space, sometimes forming a very small loop, rounded ampulla. entering intothe muscularbody wall (Figs. 12-13). Thedis- Male genital system: The deferent duct arises as a tal portion of the deferent duct is very sinuous running separate male genital duct only aftertraversing the prostate, along the right side ofthe body wall (Figs. 12-14), close to the surface and immediately below the external reproduc- tive groove which, arisingjust at side ofthe female genital pore, runs toward the base ofthe right tentacle close to the male genital system opening. At this point, the deferent duct diverges from the body wall coming into the cephalo- pedal haemocoel and turning posteriorly towardthe posteri- or end of the penis sheath, the site at which it enters the penis (Fig. 15). The penis sheath is a slender and nearly cylindrical tube (Fig. 14). The penis is slender, as long as the penis sheath; its surface is rugose due to the presence of deep transverse furrows and padded ridges or short lamel- lae. At the anterior halfofthe penis, these ridges are more or less longitudinally arranged. The prepuce is a somewhat triangular or cordiform structure with thick muscular walls (Figs. 14-15). The prepuce is shorter than the penis sheath; A its length is half or a third of that of the penis sheath. powerful penis sheath retractor muscle arises from the pos- teriorend ofthe penis sheath (Fig. 15), and it is attached at the columellar muscle. Another series offour or five mus- cularbundles are attached to the lateral wall ofthe prepuce. Two long muscularbundles arejoined to the frontal wall of the prepuce running on each side, and connecting with the penis sheath retractor. The contractionofthese two muscles determines the erectionofthe penial complex (Figs. 14-15). The deferent duct runs over the right-most muscular bun- dle, toward the proximal endofthe penis (Figs. 14-15). NERVOUS SYSTEM: The cerebral andpedal gan- glia arejoined by commissures and connectives to form the anterior nerve ring located at the anteriorhalfofthe buccal mass, just at or a little behind the origin of the esophagus (Figs. 16, 18-19). However, the location of the anterior nerve ring showed considerable variation according to the degree of retraction of the buccal mass, but it was always located at the anterior half of the buccal mass. The right pleural ganglion isjoined by somewhatshortconnectives to both cerebral and pedal ganglia. A relatively long connec- Figs. 14-17. Chilina megastoma Hylton Scott, 1958. Genital and nervous tivejoins the rightpleural ganglion to the rightparietal gan- systems.Fig. 14.Cameralucidadrawingofthedistalportionofthefemale glion (supraesophageal ganglion of Fretter, 1975), which and male genital systems. Fig. 15. Camera lucidadrawing with detailsof gives off a large nerve that supplies the osphradium (Fig. the penial complex. Fig. 16. Semi-diagramatic schemeofthe anteriorand posteriornerverings.Fig. 17. Semi-diagramaticschemeoftheinnervation 19), and a second very long and thin connective that runs to ofthe buccal mass. Scale bars = 1 mm. (dd, deferentduct; lb, left buccal the visceral ganglion (also called abdominal [Fretter, 1975] ganglion; leg, left cerebral ganglion; lpe, left pedal ganglion; lpg, left or innominate [Harry, 1964]), at the end of the posterior parietalganglion; 1pig, leftpleural ganglion; lr, lateral penisretractor; pp, nerve ring. On the left side ofthe posterior nerve ring, two prepuce; pr, penis retractor; ps, penis sheath; rb, right buccal ganglion; rcg, right cerebal ganglion; rpe, right pedal ganglion; rpg, right parietal short connectives join the left cerebral and pedal ganglia ganglion; rplg, right pleural ganglion; sig, subintestinal ganglion; v, vagi- with the pleural ganglion, from which a relatively short na;vg,visceralganglion). ITUARTE: CHIILINA MEGASTOMA 13 Figs. 18-20.ChilinamegastomaHyltonScott, 1958(MLP5210). Fig. 18. Dorsal view ofa dissected specimen to show the anterior and posterior nerverings (seeFig. 16forreference). Fig. 19. Lateral view showing the locationofthecerebralcommissure.Posteriorly,thecerebralganglion,the rightpleuralganglion(arrowhead),therightparietalganglionandthepari- etovisceralconnective,areshown.Fig. 20.Viewofthemantleroofshow- ingthe locationoftheosphradiumclose to the renal pore atthe rightend ofkidney. Scalebars= 1 mm. (cc,cerebralcommissure;eg,cerebral gan- glion; k,kidney; os,osphradium; pg,parietal ganglion; pvc,parietoviscer- alconnective;rp,renalpore). Figs.21-24.Chilinafluminea(Maton, 1809)(MLP5246).Fig.21.Dorsal view ofa partially dissected specimen showing position ofthe cerebral connective arises and runs toward the left parietal ganglion. commissure,andthelongloopsofthedeferentduct(arrowheads).Fig. 22. A long connective joins the left parietal ganglion to the Right lateral view. Fig. 23. Left lateral view. Fig. 24. Dorsal view ofthe subintestinal ganglion, which lies over the columellar mus- nervoussystem.Scalebars= 1 mm.(cc,cerebralcommissure;eg,cerebral cle, at its posteriorhalf. A somewhat large nerve arises at a ganglion; ddl, deferent duct loop; e, esophagus; fp, female pore; lb, left point approximately 2/3 the length of the left parietal- buccal ganglion; lpg, left parietal ganglion; lplg, left pleural ganglion; p, penis; pg, parietal ganglion; rpg, right parietal ganglion; sd, spermathecal subintestinal connective. There is a slight swelling just at duct; sg, salivary gland; sig, subintestinal ganglion; st, spermatheca; u, the site where that nerve originates; however, it might not uterus;v,vagina;vg,visceralganglion). 14 AMER. MALAC. BULL. 14(1) (1997) definitely be considered a ganglion. Finally, a very short downward, just at the point where the uterus connects with connective joins the subintestinal ganglion to the visceral the vagina (Fig. 21). Thereafter, to traverse the vaginaven- one, closing the posterior nerve ring. From the latter, a trally, the duct reappears dorsally and immediately opens large nerve arises, and shortly thereafter, it gives off two nearthe tip ofthe vagina. The same features (Fig. 16) have nerves which supply the visceral mass. From the subintesti- been illustrated by Harry (1964) for C. fluctuosa from nal ganglion, a large nerve runs toward the right, passing Chile. Moreover, in C. megastoma, the accessory or sec- through the columellarmuscle to innervate the distal partof ondary seminal receptacle is barely enlarged at the tip, the vagina and the pneumostomal appendage. As is charac- while in C.fluminea the tipofthis organ is greatly enlarged teristic for the genus, the pleuro-visceral connectives show into a large pear-shaped bulb. This difference in shape and incomplete torsion (Figs. 16, 18). size doesnotseem tobe relatedtoorganphysiology. At each side ofthe buccal mass (dorsolaterally), far In the male portion of the genital system, several removed, and behind the origin of the esophagus, are the differences among species should be pointed out: while in buccal ganglia. The ganglia arejoined by a somewhat short Chilina megastoma the segment ofthe deferent duct which commisure which passes below the esophagus. A nerve, runs into the muscular body wall just below the genital arising from the middle of this commisure, sinks into the groove is markedly sinuous (Figs. 12, 14), in C.fluminea it buccal mass shortly after its origin (Fig. 17). The buccal is nearly always straight or faintly sinuous (Fig. 22). In ganglia are connected to each cerebral ganglion by long addition, in C. fluminea, when the deferent duct reenters connectives, forming an U-shaped open nerve ring. The into the haemocoelic space toward the penis, it develops osphradium, located on the roofof the mantle cavity, close two or three large loops prior to running along the penial to the renal pore, and at the anterior end of the pneu- retractor muscle and entering the penis (Figs. 21-22). mostome, is a flat oval-shaped organ formed by an slightly Unlike this, in C. megastoma the deferentduct is strictly as curved furrow, bordered by two elongated and inflated lips long as the genital organs itaccompanies (Figs. 12, 14-15). (Fig. 20). According to Valdovinos and Stuardo (1995), the nervous system is the only reliable feature to differentiate Chilean species and to build suitable supraspecific arrange- DISCUSSION ments. According to Haeckel (1911), the structural pattern ofthe posterior nerve ring seems to be a specific character- From the present knowledge of the distribution of istic, especially with respect to the number of ganglia and Chilina megastoma, the species seems to be endemic to a the length of the connectives between the left pleural and reduced number of environments closely related to high subintestinal ganglia. energy freshwater courses such as vertical cliffs and rocky The general pattern of the nervous system of walls permanently swept by winds and water trickles from Chilina megastoma, here studied, does not differ from that nearby waterfalls. As reported for other gastropod species described by Harry (1964) for C. fluctuosa. However, (Trussell et ah, 1993), the extreme reduction of the spire, Harry (1964) was unable to find the left parietal ganglion the globose shape ofthe last whorl and the wide aperture of already described by Haeckel (1911). In the present study, the shell shown by C. megastoma represent adaptive the left parietal ganglion was easily identified in both C. responses to particular environmental conditions, such as megastoma and C.fluminea (Figs. 18, 23-24). In C. megas- dislodgementagents inhigh energy watercourses. toma, the lengths of the left cerebropleural connective and Hylton Scott (1958), and Castellanos and Gaillard left pleuroparietal connectives were similar. The left pari- (1981) considered that Chilina megastoma approaches C. etal-subintestinal connective was approximately three times fluminea in shell morphology, pointing to the lack of as long the pleuroparietal connective (Fig. 18). In C. flu- periostracal sculpture as adifferential characterin thelatter. minea the left pleural and parietal ganglia are closer than With respect to the anatomy of soft parts, several they are in C. megastoma,being, in some cases, nearlycon- differences should be pointed out. The general features of tiguous (Fig. 24). In C. megastoma the anterior nerve ring the genital system of Chilina megastoma are coincident tends to be located more posteriorly than in C.fluminea, in with those described by Miquel (1984) for C.fluminea and which ithas alwaysbeenobserved at the anteriorendofthe by Harry (1964) for C.fluctuosa Gray, 1828. The following buccal bulb (Figs. 21-23). differences have been observed: in C. megastoma the duct As in Chilinafluminea, the posterior end of the of the seminal receptacle or spermatheca crosses, from left radular sac of C. megastoma is projected from the buccal to right, above the lowerportion ofthe female genital com- mass, forming a short cylindrical tube bent slightly dorsal- plex, just below the floor of the pallial cavity, while in C. ly. This fact is in contrast with the description given by fluminea, the duct passes over the anterior border of the Harry (1964) forC.fluctuosafromChile. distal part ofthe spermoviduct and prostate, and then turns Several details in morphology of the radular teeth ITUARTE: CHIILINA MEGASTOMA 15 LITERATURE CITED Castellanos, Z. J. A. and M. C. Gaillard. 1981. Mollusca, Gasteropoda, Chilinidae. Fauna de Agua Dulce de la Repiiblica Argentina 15(4):20-51. Castellanos, Z. J. A. and N. A. Landoni. 1995. Mollusca Pelecypoda y Gastropoda. In: Ecosislemas de Aguas Continentales, Metodologias Para su Estudio, T. 2, E. C. Lopretto and G. Tell, eds.pp.759-801.EdicionesSur,LaPlata,Argentina. Castellanos, Z. J. A. and S. E. Miquel. 1980. Notas complementarias al genero Chilina Gray (Mollusca, Pulmonata). Neotropica 26(76):171-178. Castellanos, Z. J. A. and S. E. Miquel. 1991. Distribucion de los Pulmonata Basommatophora. Fauna de Agua Dulce de la RepiiblicaArgentina 15(9):1-11. Duncan, C. J. 1960a. The evolution of the pulmonate genital system. ProceedingsoftheZoologicalSocietyofLondon 134:601-609. Duncan, C. J. 1960b. The genital systems of the freshwater Basommatophora. Proceedings ofthe Zoological Society of London 135:339-355. Duncan, C. J. 1975. Reproduction. In: Pulmonates, Vol. 1. Functional Anatomyand Physiology, V. Fretter and J. Peake, eds. pp. 309- 365.AcademicPress,London. Fretter, V. 1975. Introduction. In: Pulmonates, Vol. 1. Functional AnatomyandPhysiology,V.FretterandJ. Peake,eds.pp.xi-xxix. AcademicPress,London. Harry, W. H. 1964. The anatomy of Chilinafluctuosa Gray reexamined, with prolegomena on the phylogeny of the higher limnic Basommatophora (Gastropoda: Pulmonata). Malacologia l(3):355-385. Haeckel, W. 1911. Beitrage zur Anatomie der Gattung Chilina. ZoologischeJahrbiicher,supplement, 13(4):89-136. Hylton Scott, M. I. 1958. Nueva especie de Chilina del norte argentino (Moll.Pulm. Basommatophora).Neotropica4(13):26-27. Miquel, S. E. 1984. Contribution al Conocimiento Bioldgico de Gasteropodos Pulmonados del Area Rioplatense, con Especial ReferendaaChilinafluminea(Maton). DoctoralThesis,Facultad Figs.25-26.RadulaofChilinafluminea(Maton, 1809).Fig.25.Detailsof central and first lateral teeth. Fig. 26. Lateral teeth. Scale bars = 25 pm de Ciencias Naturales y Museo, Universidad Nacional de La (Fig.25),50pm(Fig.26). Plata. 133pp. Miquel, S. E. 1986a. El ciclo de vida y la evolucion gonadal de Chilina flumineafluminea (Maton, 1809) (Gastropoda Basommatophora Chilinidae).Neotropica32(87):23-34. seem to differ consistently among species. In C. Miquel,S.E. 1986b.Tiposcelularesdel tractogenitalde Chilinafluminea fluminea (Maton, 1809) (Gastropoda Basommatophora megastoma, the central tooth has a definite triangular, elon- Chilinidae).Neotropica32(88):133-138. gated base, while in C. fluminea the attachment base is Trussell, G. C, A. S. Johnson, S. G. Rudolph, and E. S. Gilfillan. 1993. shorter and stronger. The lateral and marginal teeth of C. Resistance to dislodgement: habitat and size-specific differences megastoma, have more slender attachment bases and their inmorphology andtenacity inan intertidal snail. MarineEcology cusps are longer, with a more definite dagger-like shape ProgressSeries 100:135-144. Valdovinos, C. and J. Stuardo. 1995. Morfologia funcional de Chilina than those of in C.fluminea (Figs. 25-26). In addition, the angusta (Philippi, 1860), y evolucion de Chilinidae. Resumos. II last marginal tooth of each row in C. megastoma has four Congresso Latino-Americano de Malacologia, Porto Alegre, cusps (Figs. 10-11), while in C.fluminea it usually has six Brasil:43. or seven cusps (Castellanos and Gaillard, 1981). The num- ber of tooth rows also differs: 40 in C. megastoma and Dateofmanuscriptacceptance: 18June 1997 between 60 and 65 in C. fluminea (Castellanos and Gaillard, 1981).