Invertebrate Reproduction and Development, 46:1 (2004) 1- 9 Balaban, Philadelphia/Rehovot 0168-8170/04/$05 .00 © 2004 Balaban

Spermatophore morphology of the endemic Loxopagurus loxochelis (, ) from the southwestern Atlantic - and

MARCELO A. SCELZ01*, FERNANDO L. MANTELATT02 and CHRISTOPHER C. TUDGE3 1Departamento de Ciencias Marinas, FCEyN, Universidad Nacional de Mar del Plata/CONICET, Funes 3350, (B7600AYL), Mar del Plata, Argentina Tel. +54 (223) 475-1107; Fax: +54 (223) 475-3150; email: [email protected] 2Departamento de Biologia, Faculdade de Filosojia, Ciencias e Letras de Ribeirao Preto (FFCLRP), Universidade de Sao Paulo (USP), Av. Bandeirantes 3900, Ribeirao Preto, Sao Paulo, Brasil 3Department of Systematic Biology, National Museum ofNatural History, Smithsonian Institution, Washington, DC 20013-7012, USA

Received 10 June 2003; Accepted 29 August 2003

Summary The spermatophore morphology of the endemic and monotypic hermit crab Loxopagurus loxochelis from the southwestern Atlantic is described. The spermatophores show similarities with those described for other members of the family Diogenidae (especially the Cliba­ narius), and are composed of three major regions: a sperm-filled, circular flat ampulla; a columnar stalk; and a pedestal. The morphology and size of the spermatophore of L. loxochelis, along with a distinguishable constriction or neck that penetrates almost halfway into the base of the ampulla, are characteristic of this . The size of the spermatophore is related to hermit crab size. Direct relationships were found between the spermatophore ampulla width, total length, and peduncle length with carapace length of the hermit crab. These morphological characteristics and size of the spermatophore ofL. loxochelis are species-specific, distinguishing them from other members of the family Diogenidae, and can be used to infer phylogenetic relationships among them.

Key words: Spermatophore, hermit crab, Diogenidae, reproduction

Introduction there is a unique family, the Aeglidae, restricted to Anomuran dec a pods are a group of that freshwater and endemic to (Bond­ include a wide variety of forms, such as the attractive Buckup, 2003). hermit and porcellanid crabs, and some commercial According to McLaughlin (1983) and McLaughlin species such as the king crabs, among others. Some of and Holthuis (1985), the Anomura was redefined and them are semi-terrestrial, while the majority inhabits currently constitutes the following superfamilies: Gala­ marine waters, from the intertidal to the deep sea. Also, theoidea, Hippoidea, Paguroidea and Lomisoidea (see

'Corresponding author. 2 M.A. Scelzo eta!. I IRD 46 (2004) 1-9

also Martin and Davis, 2001 ). The superfamily Pagu­ Spermatophores play a major role in sperm transfer roidea, which includes the common hermit crabs and and storage in decapod crustaceans, especially in king crabs, currently consists of the Coenobitidae, hermit crabs that use a gastropod shell as a protective Diogenidae, , Parapaguridae, Pylochelidae, cover for their non-calcified abdomen. Spermatophores Lithodidae and Pylojacquesidae families (McLaughlin not only serve as protection for the non-motile sperma­ and Lemaitre, 2001; McLaughlin, 2003). The family tozoa during transmission to females but also may Diogenidae is considered to be an ancient mono­ provide energy rich substrates for prolonged sperm phyletic group (Forest, 1995), but its monophyly has storage in the females (Subramoniam, 1991 ). All the been questioned (Tudge, 1995, 1997). It is a morpho­ representatives in the Diogenidae and Paguridae are logically diverse taxon currently encompassing 19 aquatic, with mating and fertilization occurring in genera: Allodardanus, Aniculus, Bathynarius, Calci­ water. Meanwhile, members of the Coenobitidae are nus, Cancellus, , , , terrestrial (Birgus /atro) or semi-terrestrial ( Coenobita , Isocheles, Loxopagurus, , Pagu­ spp.) with mating and fertilization obviously occurring ropsis, Petrochirus, , Pseudopagu­ in the terrestrial environment (Greenaway, 2003). rus, Strigopagurus, Tisea, Trizopagurus (McLaughlin, During copulation the male hermit crab transfers the 2003). pedunculate spermatophores and attaches them to the Loxopagurus loxochelis (Moreira, 1901) is a exoskeleton of the female hermit crab or onto the species endemic to the southwestern Atlantic Ocean, surface of the temporarily inhabited gastropod shell by being one of the most common hermit crabs inhabiting the sticky base of the spermatophore pedestal (Hazlett, the shallow littoral waters of Brazil, and 1996; Hess and Bauer, 2002). The spermatophore wall Argentina, down to a depth of 30 m (Boschi et al., is molded into a variety of complex shapes by changes 1992; Melo, 1999). The biology of this species is in the contour of the vas deferens lumen, contraction of reasonably well known (Martinelli et al., 2002). The the vas deferens musculature, and deposition of mater­ data available deal only with the species description ial from adjacent glandular cells (Matthews, 1953; and adult distribution (Moreira, 1901; Forest and Fingerman, 1992; Krol et al., 1992; Tudge, 1999a). De Saint Laurent, 1967; Scelzo and Boschi, 1973; The typical, pedundulate anomuranlhermit crab Scelzo, 1976) and larval distribution along the coast of spermatophore can be divided into three major regions: Rio Grande do Sui, Brazil (Rieger and D'Incao, 1991 ). a sperm-filled ampulla, a columnar stalk of variable The life cycle of L. loxochelis has recently been length, and a foot or pedestal. The ampulla has a par­ studied in the Ubatuba region (Sao Paulo State, Brazil) tition or line of division that runs around the lateral where the occurrence of exobionts on occupied shells, edge and separates the ampulla into two halves. This shell utilization pattern, relative growth, and spatial suture line is the point of weakness where the ampulla distribution were investigated (Martinelli and Mante­ breaks to release the spermatozoa prior to fertilization latto, 1998, 1999; Mantelatto and Martinelli, 2001; (Tudge, 1991, 1999a, 1999b). According to Tudge Mantelatto et al., in press). ( 1991 , 1997), spermatophore form at both the light and Comparisons of the functional morphology of electron microscope levels can be used to separate genitalia and subsequent sperm transfer and storage species of hermit crabs within the Paguroidea and thus mechanisms among taxa provide useful information on have some phylogenetic utility. Spermatophores from phylogenetic relationships and evolutionary diver­ members of the family Coenobitidae, for example, are gence, especially in the (Bauer, 1986, 1991 ). robust in nature with large, ovoid-spherical ampullae However, particular modes of sperm transfer used by mounted on short, thick stalks. Members of the Dio­ organisms can be more directly influenced by the genidae have more fragile spermatophores with small habitat of those organisms than by the phylogenetic spherical ampullae mounted on long, slender stalks; relationships, as argued by Mann (1984). Considerable and in the Paguridae, spermatophores are distinctive in morphological variation exists among representatives possessing large, elongate, ampullae, and smaller of the reptant decapods, and members of the various accessory ampullae at the base of the main ampullae groups have evolved different strategies of transferring and a pseudo-stalk analogous with the true stalk of the sperm from the male to the female (Hinsch, 1991; Coenobitidae and Diogenidae (Tudge, 1991 ). The Subramoniam, 1991 ). Also, the mechanism by which unusual accessory ampullae have also been recorded in spermatophores are transmitted from the male gono­ some members of the Lithodidae and Parapaguridae pore to the body of a receptive female has only been (Tudge et al., 1998; Tudge, 1999a). hypothesized (see Hess and Bauer, 2002, and refer­ Since very little information is available on the ences therein). reproductive system of the endemic southwestern M.A. Scelzo et al. I IRD 46 (2004) 1-9 3

Atlantic hermit crab L. loxochelis, we describe and illustrate the spermatophore morphology and biometric relationships of this species and compare it with other members of the family Diogenidae. Any divergence in morphology and variability among the Diogenidae is noted and phylogenetic inferences furnished. ... ,I~ :a 10 PL .- Material and Methods Approximately 400 hermit crab samples were obtained in Mar del Plata/Mar Chiquita (Buenos Aires Province, Argentina) and Ubatuba (Sao Paulo, Brazil) from 1999 through 2002. The capture of the was carried out at depths between 5 and 25 m by trawl nets. Each individual was measured in shield length (SL) from the tip of rostrum to the V -shaped groove at the posterior edge of the dorsal shield with a caliper rule (0.1 mm) or stereomicroscope mounted with a graduated eyepiece. Each fresh crab was also weighed (W) using an electronic micro scale (0.001 g sensitivity). From the total number of specimens collected, almost 300 were checked for spermatophore analysis. The male reproductive system was dissected from either freshly killed or fixed specimens. An incision was made between the fourth and fifth pair of pereo­ Fig. 1. Loxopagurus loxochelis. Drawings ofspermatophores pods (P4 and P5, respectively) pulling the abdomen showing the measurements taken: TLSp, spermatophore from the rest of the cephalothorax. The thin cuticle of total maximum length; PL, peduncle or stalk length; PW, the dorsal abdomen was cut to obtain the reproductive spermatophore peduncle width; A W, spermatophore ampulla system, comprising the testis and vas deferens (VD). width; AH, spermatophore ampulla height; AD, spermato­ The distal part of the VD attached to the basal part of phore ampulla depth. P5 was specifically used for the spermatophore analy­ sis. Spermatophores obtained near the proximal part of A regression analysis was performed to compare VD, near the testis, was also observed to compare with the dimensions of the spermatophores with the size of mature spermatophore from the distal region. The the crabs. The mean, coefficient of variation (CV) dissected materials were preserved in 3% glutaral­ expressed as percentage and coefficient of determin­ dehyde or 10% formalin. ation (r) were performed for some regressions (Sokal Five to ten spermatophores from each crab were and Rohlf, 1981 ). measured and the following measurements taken (Fig. 1): spermatophore total length (TLSp) = maxi­ mum length of spermatophore from ampulla to the Results base or pedestal; peduncle or stalk length (PL) =from the base or pedestal to the neck inside the ampulla; Reproductive morphology spermatophore peduncle width (PW) = size across the There was no difference in spermatophore external peduncle; spermatophore ampulla width (A W) =maxi­ morphology between analyzed specimens from both mum width of the spermatophore ampulla; spermato­ regions (Brazil and Argentina), and therefore, the phore ampulla height (AH) = maximum height of the general information is presented with no reference to spermatophore ampulla; spermatophore ampulla depth the origin of the material. Size and wet weight of (AD)= measured from the antero-posterior axis of the hermit crabs sampled during this study ranged from 2.1 ampulla. Female and male gonopores (maximum and to 11.4 mm SL and 0.07 to 22.6 g, respectively minimum) diameter were measured for at least two (Table 1) . Paired testes are located dorsal to the specimens from every hermit size class analyzed. hepatopancreas. A coiled vas deferens from each testis 4 M.A. Scelzo eta!. I IRD 46 (2004) 1-9

Table 1. Body dimensions of hermit crab and spermatophore of Loxopagurus loxochelis

Measures Minimum Maximum Mean± SD CV(%) No of specimens SL (mm) 2.10 11.40 6.040 ± 1.520 25 .10 400 w (g) 0.07 22.60 2.970 ± 2.970 99.79 400 TLSp (mm) 0.097 0.631 0.345 ± 0.114 33.16 298 AW (mm) 0.084 0.203 0.131 ± 0.027 20.26 298 PL (mm) 0.106 0.563 0.278 ± 0.102 36.66 292 AH (mm) 0.063 0.184 0.116 ± 0.023 20.19 298 PW (mm) 0.019 0.113 0.059 ± 0.019 33.15 298

SL =shield length of hermit crab, W =wet weight of hermit crab, TLSp =spermatophore total length, A W = ampulla width, PL = peduncle length, AH = ampulla height, PW = peduncle width. connects them to the base of the 5th pair ofpereopods. found at the distal part of the peduncle, near the neck Male and female gonopores are operculate, and exhibit (Fig. 2D). an ovoid shape. The male gonopores are surrounded by The ampulla width showed a low correlation with plumose and serrate setae; but the female gonopores hermit crab size (AW = 0.0172 SL + 0.0192, r= 0.24), are almost free of setation. The maximum mean dia­ but was well correlated with ampulla length (A W = meter of female gonopores measured 390 ± 92 Jlm 0.7922 AH + 0.0392, r= 0.49), and to spermatophore (mean± SD) but ranged from 200 to 560 Jlm for crabs total length (A W = 0.1604 TLSp + 0.0759, r = 0.48). measuring 3.8-6.9 mm SL. Male gonopores measured The peduncle length is also directly related to sperma­ a mean of 343 ± 107.6 Jlm (range 160-510 Jlm) for tophore total length (PL = 1.0454 TLSp + 0.0574, r= crabs measuring 5-11.4 mm SL. A direct relationship 0.90). Each spermatophore has a distinct dorso-lateral was obtained between the large diameter of gonopore ridge or suture obvious as a line around the ampulla. and hermit crab size (male gonopore diameter= 51.548 SL, 71.52, r= 0.82,N= 31; female gonopore diameter = 68.006 SL, 1.8337, r= 0.98, N= 20). Discussion L. loxochelis is gonochoric and sexually dimorphic, Spermatophore morphology with males growing more and faster than females, and both sexes reaching sexual maturity at the size 4.5 mm Spermatophores of L. loxochelis form a coiled line SL (Mantelatto and Martinelli, 2001 ). In Brazilian united by the semi-circular-shaped pedestal in almost waters (Ubatuba) ovigerous females occur all year with all individuals analyzed. The spermatophores are flattened antero-posteriorly and they have three major some peaks in the winter and spring (Martinelli et al., regions: a sperm-filled, circular flat ampulla, a colum­ 2002). Meanwhile reproduction in Argentina (Mar del nar stalk and a pedestal (Figs. 1 and 2). Spermatophore Plata and Mar Chiquita) takes place during spring­ total length was independent of the hermit crab size, summer when larvae are released (MAS, unpublished measuring between 97-630 Jlm TLSp (CV = 33.16%). results). L. loxochelis has three zoeae stages and one The ampulla is generally circular in shape and flat­ megalopa or glaucothoe during larval development tened antero-posteriorly, measuring between 84 to (Bernardi, 1986). 203 Jlm AW (CV = 20.26%), 63 to 184 J.Lm AH (CV = The absence of differences in spermatophore 20.19%) and 60 Jlm AD antero-posteriorly. In some morphology between individuals was predictable con­ individuals the external shape of the ampulla was sidering that preliminary molecular analysis suggests found to be more rectangular or square. The smooth that Loxopagurus is a valid monotypic southern genus, peduncle penetrates half way into the spermatophore and that no marked genetic variability occurs among head and is between 106 to 563 Jlm PL in length (CV = populations of the species from Brazil and Argentina 36.66%), and 19 to 113 J.Lm PW in width (CV = (Mantelatto et al., 2002). 33.15%). The peduncle shows great variation in length Similar to other diogenid species, the sperma­ and in shape (Table 1), and so it is not considered a tophore morphology of L. loxochelis is consistent with useful measurement for comparisons between indivi­ the pedunculate spermatophore type common in the duals. In some individuals, at the proximal part of the Coenobitidae and Diogenidae and even some species vas deferens, two-headed (two ampullae) sperma­ of Paguridae (Tudge, 1991, 1999a; Tudge and tophores were found (Fig. 2C). A subapical knob was Jamieson, 1991 ). This morphology is most like the M.A. Scelzo eta/. I IRD 46 (2004) 1-9 5

100 Jlffi 100 Jlffi 100 Jlffi A B c

I I

I I I I I I I I I

I I I I I I I I I I I

I I 100 Jlffi 50Jlm 100 Jlffi I l J..--...:.-.f

I D E F

Fig. 2. Loxopagurus loxochelis. Light and electronic micrographs of spermatophores. A, spermatophore; B, ampulla; C, double-headed ampulla; D, peduncle with subapical knob (frontal view); E, SEM image of head spermatophore showing escaping spermatozoan from dorsal suture line; F, semicircular pedestal.

Clibanarius genus (Mouchet, 1930, 1931; Hamon, within the limits of a characteristic familial mor­ 1942; Tudge, 1991) especially C. corallinus, C. misan­ phology. With some exceptions, the spermatophores thropus, and C. virescens, but less so like the larger, generally have long, thin stalks (less than 20 t-tm in stalk-less spermatophores of C. longitarsus and C. vit­ diameter) and small spherical ampullae approximately tatus (Uma and Subramoniam, 1984; Matos et a!., 50-70 t-tm in size (Tudge, 1991 ). 1993; Hess and Bauer, 2002). Similarities occur in the Tudge (1991) also presented a practical branching overall morphology but also in the small posterior key to classify the hermit crabs into their respective pointing projections on the underside of the ampulla. families based on the structure of their spermatophores. This phylogenetic similarity between the sperma­ Our findings agree with this study in which light thophore morphology ofLoxopagurus and Clibanarius microscope observations of spermatophores can be is in accordance with the recent molecular phylo­ used successfully to distinguish hermit crab families of genetic results obtained for the Diogenidae species the Paguroidea, especially within the Diogenidae. The from the South Atlantic (FLM, personal observation). spermatophore morphology of hermit crabs in the Studies on other members of the family Diogenidae Diogenidae has been described for 21 species in nine show a great diversity of spermatophore forms but genera, including the present study (Table 2). 6 M.A. Scelzo et al. I IRD 46 (2004) 1-9

Table 2. List of species ofhermit crab of the family Diogenidae with published spermatophore descriptions and/or illustrations

Species References Aniculus maximus (as Trizopagurus maximus) Matthews, 1957 latens Tudge, 1991 Calcinus minutus Tudge, 1991 Calcinus ornatus Hamon, 1942 Ciliopagurus strigatus (as Aniculus strigatus) Matthews, 1957 Clibanarius cora/linus Tudge, 1991 Clibanarius longitarsus Uma and Subramoniam, 1984 Clibanarius misanthropus Mouchet, 1930, 1931; Hamon, 1942 Clibanarius virescens Tudge, 1991 Clibanarius vittatus Matos eta!., 1993; Hess and Bauer, 2002 Dardanus arrosor (as Pagurus arrosor) Mouchet, 1931; Hamon 1939, 1942 Dardanus asper Matthews, 1953 (as Pagurus callidus) Hamon, 1942 Dardanus lagopodes Tudge, 1991 Tudge, 1991 Dardanus punctulatus Matthews, 1956 Diogenes gardineri Tudge, 1991 Mouchet, 1930; Hamon, 1942; Tuzet and Manier, 1961; Manjon-Cabeza and Garcia Raso, 2000 Loxopagurus loxochelis Present study Paguristes oculatus Mouchet, 1931; Hamon, 1942 Strigopagurus boreonotus Tudge, 1996

Table 3. Morphometry (J.Lm) of the spermatophore of several described hermit crabs of the family Diogenidae

Species* TLSp PL AH AW PW

Calcinus minutus 180 110 60 70 10 Calcinus latens 150 110 50 50 15 Clibanarius virescens 150 70 75 75 10 Clibanarius cora/linus 300 220 60 75 15 Dardanus lagopodes 600 530 70 80 20 Dardanus megistos 160 90 60 45 20 Diogenes gardineri 430 300 130 130 15 Loxopagurus loxochelis 345 280 115 130 60 Strigopagurus boreonotus 850 170 220

*Except for L. loxochelis, all data were obtained and calculated from Tudge (1991, 1996). TLSp, spermatophore total length; PL, peduncle length; AH, ampulla height; A W, ampulla width; PW, peduncle width. See Table 2 for origin of the data.

Spermatophore morphometry could be one criterion total length almost half of this length (300 J.Lm). Also, to distinguish species of diogenid hermit crabs. In D.lagopodes have an ampulla measuring (70x80 J.Lm), Table 3, selected species of diogenid hermit crabs are much smaller than L. loxochelis (115x 130 J.Lm). Some compared on this point. C/ibanarius corallinus, Dar­ individuals of D. lagopodes spermatophore have been danus lagopodes, Diogenes gardineri and L. loxochelis recorded with spermatophores possessing two ampul­ present a spermatophore mean total length greater than lae at the apex of a single stalk or peduncle (Tudge, 300 J.Lm and can be separated from the remaining 1991 ). This condition was also recorded in some described species, which have smaller spermatophores. spermatophores of L. loxochelis located at the proxi­ Diogenes gardineri and L. loxochelis share similar mal part of the vas deferens. In both D. lagopodes and ampulla size but are differentiated by the wider pedun­ L. loxochelis the double-headed spermatophores were cle of L. loxochelis. Spermatophores of Dardanus rare, and most probably represent abnormalities in lagopodes have a long peduncle (600 J.Lm in total spermatophore development and are not directly length); meanwhile, those ofL. loxochelis have a mean associated with maturity of the spermatophores. M.A. Scelzo et al. I IRD 46 (2004) 1-9 7

Clibanarius cora/linus presents a similar spermato­ Invertebrate Zoology, National Museum of Natural phore total length (Tudge, 1991) when compared with History, Smithsonian Institution, Washington, D.C., is L. loxochelis, but can be differentiated by the ampulla gratefully acknowledged. dimensions, that are smaller in C. cora/linus (60 x 75 JLm) than L. loxochelis (115x130 JLm). In many cases no accurate measurements are available for many References diogenids investigated for spermatophore morphology Bauer, R.T., Phylogenetic trends in sperm transfer and (e.g., Matthews, 1953, 1956, 1957). There still remain storage complexity in decapod crustaceans. J. Crust. many genera and species ofDiogenidae (Allodardanus, Bioi., 6 (1986) 313- 325. Bathynarius, Cancellus, Isocheles, Paguropsis, Petro­ Bauer, R.T., Sperm transfer and storage structures in chirus, Pseudopaguristes, Pseudopagurus, Tisea and penaeoid shrimps. A functional and phylogenetic Trizopagurus) for which spermatophore morphology is perspective. In: Sexual Biology, Bauer, R.T. unknown, and their description and comparison are and Martin, J.W. (eds.), Columbia University Press, New topics for future studies. York, 1991, pp.183-207. Bernardi, J.V.E., Desenvolvimento larval de Loxopagurus The morphology and size of the spermatophore of loxochelis (Moreira, 1901) (Crustacea, Decapoda, Dio­ L. loxochelis, along with a distinguishable constriction genidae) em laborat6rio. Bachelor Thesis, Universidade or neck that penetrates almost halfway into the base of Estadual Paulista (UNESP), Rio Claro, Brazil, 1986, the ampulla, are characteristic of this species. L. loxo­ pp. 1-36 (unpublished). chelis is very close to species of the genus Isocheles in Bond-Buckup, G., Familia Aeglidae. In: Manual de Identi­ terms of general somatic morphology (Forest and Saint ficayao dos Crustacea Decapoda de agua doce do Brasil, Laurent, 1967) and molecular sequence (Mantelatto et Melo, G.A.S. (ed.), Editora Loyola, Sao Paulo, 2003, a!., 2002). Both species also present similarities in pp. 21-116. general external shape of the spermatophore, but with Boschi, E.E., Fischbach, C.E. and Iorio, M.I., Catalogo Ilus­ trado de los Crustaceos Estomat6podos y Decapodos minor differences in the peduncle (FLM and MAS, Marinos de Argentina. Frente Maritimo, Vol. 10, Monte­ unpublished results). A future, more accurate analysis video, Uruguay, 1992, pp. 1- 94, Figs. 1- 98. involving ultrastructure of spermatophores will help to Fingerman, M., Glands and secretions, In: Microscopic Ana­ clarify this relationship. tomy of Invertebrates, Harrison, F.W. and Humes, A. G. Determining phylogenetic relationships based on (eds.), Vol. 10, Decapod Crustacea, Wiley-Liss, New spermatophore description alone is premature; but with York,I992, pp. 245-394. the description of spermatophores from more diogenid Forest, J., Crustacea Decapoda Anomura: revision du genre species, as well as other taxonomic criteria, such as Trizopagurus Forest, 1952 (Diogenidae), avec l'etab­ lissement de deux genres nouveaux. In: Resultats des cladistic analysis, molecular sequences, genetic, larval Campagnes MUSORTOM, Croisnier, A. (ed.), Mem. morphology, ultrastructure of sperm morphology and Mus. Nat. Hist. Nat., Vol. 163, 1995, pp. 9-149. evidence from the fossil record (Martin and Davis Forest, J. and Saint Laurent, M., Crustaces Decapodes: 2001 ), it may be possible to determine the phylogeny Pagurides. Campagne de Ia Calypso au large des cotes and evolution among the anatomically diverse family Atlantiques de I' Amerique du sud (1961- 1962). 6. Crus­ Diogenidae, as well as within the Anomura. taces Decapodes: Pagurides. Ann. Inst. Oceanogr., 45(2) (1967)47- 170. Greenaway, P., Terrestrial adaptations in the Anomura (Crustacea: Decapoda). In: Lemaitre, R., and Tudge, Acknowledgements C.C. (eds.), Biology of the Anomura. Proc. Symposium at the Fifth International Crustacean Congress, This study was supported in part by the following Melbourne, , 2001. Mem. Mus. Victoria, 60(1) Grants: PIP CONICET 4336/02, Universidad Nacional (2003) 13-26. de Mar del Plata, EXA 207/02 (Argentina). FLM and Hamon, M., La constitution chimique des spermatophores de MAS are grateful to CNPq - Prosul Program (Grant Crustaces superieurs du groupe des pagurides. C.R. Soc. #170038/02-5) for financial support during Brazil and Bioi., 130(1939) 1312- 1315. Argentina visiting program. Special thanks to those Hamon, M., Recherches sur Ies Spermatophores. Theses persons who collaborated during the development of presentees a Ia faculte Sciences de L'Universite d' Alger, Maison-Carree, Alger, 1942, p. 185. this study, especially to Eng. Jorge Martinez-Area for Hazlett, B.A., Reproductive behavior of the hermit crab aid in graphic preparation, Lie. Nestor M. Lucero and Clibanarius vittatus (Bose, 1802). Bull. Mar. Sci., 58 Dr. Renata Biagi for helping during processing of the (1996) 668-674. material and to fisherman Jose. L. Ungarelli and Benito Hess, G.S. and Bauer, R.T., Spermatophore transfer in the Mattera for fishing activities in Argentina. The support hermit crab Clibanarius vittatus (Crustacea, Anomura, of Rafael Lemaitre and Dave Pawson to CCT in Diogenidae). J. Morpho!., 253 (2002) 166- 175. 8 M.A. Scelzo eta/. I IRD 46 (2004) 1-9

Hinsch, G.W., Structure and chemical content of the sper­ Matthews, D.C., The probable method of fertilization in matophores and seminal fluid ofreptantian decapods. In: terrestrial hermit crabs based on a comparative study of Crustacean Sexual Biology, Bauer, R.T. and Martin, J.W. spermatophores. Pac. Sci., 10 (1956) 303-309. (eds.), Columbia University Press, New York, 1991, Matthews, D.C., Further evidence of anomuran non­ pp. 290-307. pedunculate spermatophores. Pac. Sci., 11 (1957) 380- Krol, R.M., Hawkins, W.E. and Overstreet, R.M., Repro­ 385. ductive components, In: Microscopic Anatomy oflnver­ McLaughlin, P.A., Hermit crabs - are they really poly­ tebrates, Harrison, F.W. and Humes, A.G. (eds.), phyletic? J. Crust. Bioi., 3 (1983) 608-621. 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