MALACOLOGIA, 2004, 46(1): 127−156

TOWARD COMPREHENSIVENESS: INCREASED MOLECULAR SAMPLING WITHIN AND ITS PHYLOGENETIC IMPLICATIONS

Christopher P. Meyer

Florida Museum of Natural History, University of Florida, Gainesville, Florida 32611 USA; [email protected]

ABSTRACT

This paper introduces 73 additional taxa to the existing mitochondrial molecular data- base of 202 taxa for the Cypraeidae and addresses the systematic implications of their inclusion. Five outgroup members from the are also added. Sequence data are included from all previously missing extant named genera (, and ), completing the overall “generic-level” framework for living . Newly added taxa include 47 recognized , 25 subspecies, and six undescribed taxa. Phy- logenetic results generally are consistent with previous arrangements, with few minor ad- justments. The most significant findings are that: (1) currently recognized Nesiocypraea is broken into two disparate clades, a deeply rooting Nesiocypraea sensu stricto group and the more derived Austrasiatica (Lorenz, 1989). (2) Two newly included Barycypraea taxa are sister to , reaffirming the validity of the subfamilial clade Bernayinae. (3) The inclusion of a significant number of added Erroneini taxa (N = 24) creates a phylogenetic challenge because of poor support and recovered relationships inconsistent at first glance with traditionally recognized affinities. In order to maintain nomenclatural consistency, is maintained at a generic level, whereas Adusta is dropped to subgeneric status within Erronea. Greater than 90% of currently recognized species are included, and 93% of these are supported by molecular criteria. Moreover, more than 70% of the tested, recognized subspecies are distinct. The phylogeny provides one of the most comprehen- sive, species-level frameworks to date for testing diversification theories in the marine tropics. Key words: Cypraeidae, molecular systematics, taxon sampling, .

INTRODUCTION These phylogenetic hypotheses of relationship establish sister pairs at the appropriate taxo- Cowries (: Cypraeidae) are taxo- nomic level and provide the framework to test nomically one of the best known of all mollus- diversification theories. Meyer (2003) introduced can groups, and have been used frequently to molecular data for 234 taxa in Cypraeidae and examine speciation and biogeographic pat- generated phylogenetic hypotheses for most terns in the marine tropics (Schilder, 1965, major clades as well as sister-group relation- 1969; Foin, 1976; Kay, 1984, 1990; Meyer, ships for most species. Systematics for 2003). A wealth of taxonomic (Schilder & Cypraeidae were reviewed in light of the results Schilder, 1938, 1971; Schilder, 1939; Lorenz & and diversification patterns within the tropics Hubert, 1993; Groves, 1994; Lorenz, 2002), were addressed. The study presented herein anatomical (Troschel, 1863; Vayssière, 1923, significantly increases the comprehensiveness 1927; Riese, 1931; Risbec, 1937; Schilder, of taxon sampling in the group by introducing 73 1936; Kay, 1957, 1960, 1963, 1985, 1996; Cypraeidae and five Ovulidae taxa to the exist- Bradner & Kay, 1996; Lorenz, 2000), biogeo- ing molecular dataset and discusses their sys- graphic (Schilder, 1965, 1969; Foin, 1976; tematic implications. In addition to broader Burgess, 1985; Liltved, 1989; Lorenz & taxonomic sampling, this paper presents the re- Hubert, 1993; Lorenz, 2002) and fossil data sults of broader geographic sampling. The ap- (Schilder & Schilder, 1971; Kay, 1990, 1996; pendix lists 147 localities added across the Groves, 1994) is available for the group; how- various taxa. Five outgroup taxa from six locali- ever, what has been lacking is a well-resolved, ties are included, and 67 recognized cypraeid comprehensive species-level phylogeny. species or subspecies are added from 75 locali- 127 128 MEYER ties. The remaining 66 localities were added to Molecular Methods supposedly known taxa, but revealed six previ- ously unrecognized taxa, some of which may Most methods follow protocols detailed in correspond to names currently in synonymy Meyer (2003) for all aspects of preservation, upon review of type localities. extraction, amplification, and sequencing. Tis- sue samples were acquired from a variety of

MATERIALS AND METHODS

Recognition Criteria: ESU versus OTU

The ultimate goal of this project is to con- struct a comprehensive phylogeny of cypraeid gastropods at the appropriate level for diversi- fication studies. As such, the operational taxo- nomic unit (OTU) chosen for phylogenetic analyses generally represents an evolution- arily significant unit (ESU) that must fulfill some minimal criteria established through genetic scrutiny. First, mtDNA haplotypes of sampled individuals must represent a mono- phyletic clade; yet this alone is not sufficient, because any phylogeny has a plethora of monophyletic groups, because a clade re- quires only two individuals. Thus, auxiliary cri- teria are required to delineate significant units. Within cowries, these additional criteria are (1) geographic distinction or allopatry, (2) signifi- cant genetic distance from the sister group such that pairwise distance comparisons yield a bimodal distribution, and/or (3) taxonomic recognition by previous workers. An OTU is included in analyses only if at least two of these three criteria are met. Most OTUs fulfill all three criteria and are considered evolution- arily significant units (ESUs) (sensu Moritz, FIG. 1. ESU vs. OTU criteria. Phylogram show- 1994). These criteria are erected in order to ing the relationships among members of the Pacific subclade, with bootstrap val- delineate independent evolutionary trajecto- ues for major groups. Four distinct clades are ries, but do not guarantee that the units are evident, and the names presented on the right: reproductively isolated. In a few instances, two Cribrarula catholicorum, C. gaskoini, C. astaryi, of the three criteria (genetic separation and and C. cumingii. Note that single individuals of taxonomic recognition) are not supported by two newly included taxa, C. taitae and C. garciai the third (exclusive geographic signatures). (white stars), nest within two of the major clades and show little variation (a single mutation). While the genetic differences (monophyly) These two new taxa are introduced as OTUs, between populations indicate some indepen- because of their distinct morphology and geog- dent period of evolutionary history between raphy (American Samoa and Easter Island, re- geographic regions, it appears that, on occa- spectively), but are currently not considered sion, haplotypes from outlying regions can mix ESUs by molecular criteria. All individuals from back into the sister gene pool. The few cases the Marquesas are C. astaryi; however, two in- dividuals of C. cumingii possess haplotypes be- where all three criteria are not fulfilled always longing to the C. astaryi clade as well (dark stars). occur on the periphery of regions (e.g., While the two haplotype clusters are distinct, the Marquesas, ) and show asymmetrical, pattern indicates uni-directional exchange of lar- “downstream”, dispersal events (Fig. 1). As vae downstream from the Marquesas (C. astaryi). circumscribed, all ESUs discussed indicate Molecular criteria recognize these two clades as ESUs with historically limited exchange. (TIK = independent evolutionary histories, but alter- Tikehau, RR = Rangiroa, HUA = Huahine, all C. native criteria, such as either nuclear markers astaryi from Marquesas, all C. gaskoini from or breeding experiments, are needed to verify Hawaii, and all C. catholicorum from Solomon reproductive isolation. Islands) INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 129 sources and locations (listed in the Phylogenetic Analyses acknowledgements and appendix). Most samples were preserved in 95% ethanol. DNA The 297 operational taxonomic units (OTUs) extraction was performed using DNAzol presented in this paper were selected from an (Chomczynski et al., 1997) using one-half vol- extensive database comprised of over 2,000 umes and following the manufacturer’s proto- sequenced individuals. In general, taxa are col (Molecular Research Center, Inc.) with the included if they exhibit distinctive geographic exception that the digestion step was in- and/or genetic signatures. In most instances, creased by an additional 24 or 48 h. PCR was new OTUs are recognized in the literature as performed as described in Meyer (2003). COI either species (N = 47) or subspecies (N = primers were as follow (from Folmer et al., 25). This paper introduces six previously un- 1994): LCO-1490 (5’−3’) GGT CAA CAA ATC recognized taxa. ATA AAG ATA TTG G, and HCO-2198 (5’−3’) The increasing size of this dataset presents TAA ACT TCA GGG TGA CCA AAA ATC A. computational and heuristic challenges for For problematic taxa, these primers were de- phylogenetic analyses. Two weighted trans- generated as follows: dgLCO-1490 (5’−3’) version bias parsimony searches (3:1 and 5:1) GGT CAA CAA ATC ATA AAG AYA TYG G, and were performed on the complete dataset us- dgHCO-2198 (5’−3’) TAA ACT TCA GGG TGA ing PAUP* (Swofford, 1998). At first, 250 ran- CCA AAR AAY CA. Two internal primers were dom-addition replicate searches were designed for small amplifications of degraded performed, but with a tree limit of ten imposed DNA: InCypLCO (5’−3’) CGT YTA AAT AAT to minimize search time on suboptimal islands. ATA AGY TTY TG, and InCypHCO (5’−3’) CGT After 250 replicates, the most parsimonious ATA TTA ATA ATT GTT GTA AT. Palumbi’s topologies were used as starting trees for ex- (1996) 16Sar and 16Sbr primers were used haustive searches without tree limits. This for 16S: 16Sar (5’−3’) CGC CTG TTT ATC AAA strategy was employed for both weighted AAC AT, and 16Sbr (5’−3’) CCG GTC TGA ACT analyses, and the most parsimonious topolo- CAG ATC ACG T. Two internal primers were gies were pooled and evaluated using likeli- designed for small amplifications of degraded hood criteria. ModelTest v. 3.06 (Posada & DNA: In16Sar (5’−3’) GGG CTA GTA TGA ATG Crandall, 1998) was used to select the most GTT TGA, and In16Sbr (5’−3’) ATG CTG TTA appropriate model for likelihood parameters. TCC CTA TGG TAA CT. The polymerase chain The most likely weighted parsimonious trees reaction was carried out in 50 µl volumes, us- were then compared using consensus meth- ing 1 µl of template. Each reaction included 5 ods. µl 10X PCR buffer, 5 µl dNTPs (10mM stock), A two-tiered, compartmentalized strategy

2 µl of each primer (10µM stock), 3 µl MgCl2 was adopted that followed Meyer (2003) for solution (25 mM stock), 0.2 µl Taq (5 Units/µl levels of topological support. The strict con- stock) and 31.8 µl ddH2O. Reactions were run sensus topology derived from the most likely for 35-40 cycles with the following parameters: overall analyses was divided into four an initial one min denaturation at 95°C; then subequal components called basal, mid1, cycled at 95°C for 40 sec (denaturation), 40°C mid2, and derived. Because the basal, mid1 to 44°C (COI) or 50°C to 54°C (16S) for 40 sec and mid2 cohorts are necessarily paraphyletic (annealing), and 72°C for 60 sec (extension). groups that include the common ancestor and Successfully amplified products were cleaned some, but not all, of its descendants, repre- for cycle sequencing using Wizard PCR Preps sentative derived clades were included in the (Promega). Sequencing also followed Meyer paraphyletic analyses. In this way multiple (2003) with all new sequences generated using derived member clades overlapped between ABI chemistry and sequencers. Sequences were more basal and derived analyses, and the generated from the resulting electrophenograms overall topology could be “scaffolded” together using Sequencher (Gene Codes). by linking clades shared in both basal and All primer sequences, aligned COI and 16S derived compartments. sequences and Nexus files are available at Within each of the four subanalyses, parsi- the archived data web pages of the Florida mony searches were performed using a 5:1 Museum of Natural History Malacology Depart- transversion bias. Both bootstrap (Felsenstein, ment (http://www.flmnh.ufl.edu/malacology/ 1985) analyses (1,000 replicates) and decay archdata/Meyer2004), and new sequences are (Bremer, 1994) analyses (TreeRot v2; deposited in Genbank under accession num- Sorenson, 1999) were performed to establish bers AY534351 through AY534503. levels of support. Results from Bayesian meth- 130 MEYER ods (Mr. Bayes v3.04b) are not reported in this ferent. In 3:1 topologies is more paper, but were generated for the four sub- basal, while in 5:1 trees, Erronea is more basal. groups and compared to the combined parsi- When alternative topologies were evaluated mony/likelihood methods utilized in PAUP*. using ModelTest, the GTR+I+G model was se- Overwhelmingly, they were consistent with the lected as the best-fit model. When both the 3:1 results presented here, but on few occasions MPTs and 5:1 MPTs were evaluated using the differed in hypotheses of relationship. The selected likelihood criteria [lset base = scaffolded parsimony global topologies were (0.315128 0.136452 0.111915), Nst = 6, compared to the scaffolded Bayesian topol- Rmat = (0.99559 41.36057 1.0461 1.68935 ogy using likelihood criteria in PAUP*. The 22.78834), rates = gamma, shape = 0.562423, combined topology derived from the compart- Pinvar = 0.48426], the 5:1 subset was signifi- mentalized Bayesian subsets was less likely cantly more likely (ANOVA: p < 0.001, average than the overall topologies found using the –ln likelihood = 49513.8). Therefore, results combined parsimony/likelihood criteria. It ap- from the 5:1 searches are presented herein. pears that Bayesian results depended on taxon The overall relationships among major sub- sampling and outgroup inclusion. While this groups recovered in the 5:1 MPTs are more finding may be of interest to the general sys- consistent with both morphological and fossil tematic community, it is not a point specifically evidence in addition to being more likely based addressed in this paper. on molecular data. In particular, a monophyl- etic pustulose clade is more parsimonious for conchological and anatomical features, be- RESULTS cause it is more likely that a bumpy shell was derived a single time, rather than being derived The final culled dataset contained 297 OTUs either twice independently, or derived once then and 1,107 characters, 493 base pairs from 16S lost. Also, the basal, paraphyletic status of and 614 bases from COI. For 16S, alignment and within the 3:1 followed those presented in Meyer (2003) topologies is inconsistent with the fossil record based on secondary structure. Weighted par- for both groups relative to more derived mem- simony searches resulted in 512 equally most bers of the 3:1 MPTs (i.e., , Barycypraea, parsimonious trees (MPTs) for 3:1 Ti:Tv and 480 and Zoila), which appear earlier in the record trees for 5:1 searches. Derived portions of the and root more deeply in the 5:1 topologies. Also, comprehensive topology were consistent. Thus, the sister-group relationship of the two genera all named clades (subfamilies, tribes and gen- is more consistent with paleobiogeography (the era) presented in Figure 2 are found in all to- breakup of Gondwanaland) and recognized pologies, except one mentioned below. affinities based on both conchological and de- However, the topologies recovered from alter- velopmental criteria. The other major discrep- nate weightings differed in five deeper regions, ancies between the 3:1 and 5:1 MPTs (most all of which are poorly supported regardless of basal cowries, Perisserosa affinities, and posi- methodology. First, 5:1 topologies placed the tion of Purpuradusta) are more ambiguous clade consisting of Propustularia/Nesiocypraea/ based on alternate criteria (morphological or basal as sister to all other cowries. In 3:1 paleontological). topologies this clade moves up one node and is sister to Erosariinae. Second, the pustulose Suprageneric Relationships (Fig. 2) clade consisting of Nucleolaria/Cryptocypraea/ is monophyletic in 5:1 trees, while Overall, suprageneric results were consistent in 3:1 topologies these genera are a basal with previous systematic findings (Meyer, paraphyletic grade leading to the clade includ- 2003), with two exceptions. First, Ipsa falls out- ing /Perisserosa/Erosaria. Third, in side Erosariinae and is no longer sister to 5:1 topologies Perisserosa is sister to Erosaria, Erosariini, but instead is allied with newly in- whereas in 3:1 trees, Perisserosa is sister to cluded Propustularia and Nesiocypraea sensu Monetaria. Fourth, the arrangement of major stricto. New sequence data from Nesiocypraea groups along the backbone from Umbiliini to teramachii neocaledonica did not result in an Cypraeovulinae conflicts. Results from 5:1 affinity with other recognized “Nesiocypraea” searches are shown in Figure 2, whereas in species (N. hirasei, N. sakurai and N. langfordi). 3:1 topologies, Notocypraea and Cypraeovula Instead, Nesiocypraea teramachii roots more (Cypraeovulinae) are a basal sister grade lead- deeply in the phylogeny as a distant sister to ing to more derived member groups. Finally, , within a clade that includes both the basal arrangement within Erroneini is dif- Ipsa and Propustularia. Thus, the inclusion of INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 131

FIG. 2. Strict suprageneric consensus topology of 480 most parsimonious trees derived from a 5:1 Ti:Tv weighted search strategy of all 297 OTUs. Subfamilies are indicated with arrows and tribes are listed to the right. The four compartments for further subanalyses are bracketed to the right. The four newly added genera are capitalized and bolded. 1Lyncina includes the subclades , Miolyncina and as reported in Meyer (2003). 2Austrasiatica replaces the prior use of Nesiocypraea for the same clade. 3Erronea now includes Adusta, formerly rec- ognized as the sister taxon. 132 MEYER two new ancient lineages (Propustularia and and Zonaria was a surprising Nesiocypraea) affects the relative position of result (Meyer, 2003). These new data for Ipsa. Moreover, the finding that Nesiocypraea Schilderia place the as sister to Zonaria teramachii is not related to other previously rec- to the exclusion of Pseudozonaria (and ognized Nesiocypraea, compels me to recog- ), and phylogenetic results main- nize the clade Austrasiatica proposed by Lorenz tain their independent, paraphyletic status. (1989) at the generic level for the group includ- These finding are more consistent with geo- ing Austrasiatica hirasei, A. sakurai, and A. graphic affinities than recognized taxonomic langfordi. There are some conchological and affinities (Pseudozonaria is often considered a anatomical features that support this separa- subgenus of Zonaria), as both Neobernaya and tion. The left posterior terminal ridge in Pseudozonaria are currently restricted to the Nesiocypraea is more produced and separate eastern Pacific whereas Schilderia and Zonaria from the body of the shell, whereas in are restricted to the western Atlantic. Austrasiatica, the ridge is continuous with the body. Lorenz (pers. comm.) also states that (1) Basal Compartment (Fig. 3) Nesiocypraea lacks a distinct embryonic band- ing, having instead only a darker middorsal Five Ovulidae taxa are added in these analy- zone, (2) Nesiocypraea have a proportionally ses: Pseudocypraea exquisita, volva, larger , and (3) the darker pattern of the concinna, takeoi, and shell is absent in juvenile Austrasiatica, only semperi. Within Ovulidae, only a few gained after the deflection of the labral margin; major clades are well supported and may be whereas, the darker pattern can be part of ju- the results of poor taxon sampling. First, the venile Nesiocypraea shells. Additionally, the clade Eocypraeinae appears well supported rachidian tooth of Nesiocypraea lacks the and includes Pedicularia, and prominent paired basal denticles present in the Pseudocypraea. Eocypraeinae is sister to a three Austrasiatica taxa, and the tooth shape strongly supported clade (Ovulinae) that in- is less elongated and squared, whereas the cludes the remaining Ovulidae. Within the rachidian in Austrasiatica narrows toward the Ovulinae, two subgroups are well supported cusps (Bradner & Kay, 1996). The fact that and represent the major clades Volvini and Austrasiatica was erected to differentiate the Ovulini. Of the added Ovulidae, Volva falls into three species (albeit incorrectly aligned with Volvini, but Prosimnia unexpectedly falls into Schilderia) is also an indication that the two lin- Ovulini as do Primovula and Dentiovula. These eages possess independent histories. The deep results are generally consistent with Cate’s position of Propustularia within the phy- (1974) arrangement of higher-level relation- logeny is not surprising because it is one of the ships within the Ovulidae. Cyphoma gibbosum oldest of extant taxa, extending back to the falls basal to these two sisters in the strict con- Lower Eocene (Kay, 1996). sensus topology; however, its position is poorly The second suprageneric difference concerns supported, and it is expected to move within the relative position of Zoila in the overall phy- the Volvini with the inclusion of more taxa. logeny and is caused by the inclusion of se- Monophyly of Ovulidae is not addressed herein quence data for two taxa from the ancient and would require the inclusion of more distant lineage Barycypraea. These new data indicate representatives from Lamellaridae, that Barycypraea teulerei and Barycypraea and . fultoni are sister taxa, and they are sister to The Cypraeidae basal group includes the Zoila. This Barycypraea/Zoila clade is recog- genera Propustularia, Nesiocypraea, Ipsa, nized as the extant members of the subfamily Cryptocypraea, Nucleolaria, Staphylaea, Bernayinae, a group that includes many extinct Monetaria, Perisserosa, and Erosaria. fossil members and extends back into the Me- Propustularia, Nesiocypraea, and Ipsa form a sozoic (Kay, 1996). These new data change clade that roots deeply within the phylogeny and the relative position of Zoila to Cypraeinae is sister to all other cowries. Each of the three (Meyer, 2003); however, the topology in this genera is represented by only a single taxon, region of the phylogeny is poorly supported. and only Nesiocypraea contains additional rec- The final suprageneric addition to the molecu- ognized species missing from the dataset lar database is the inclusion of sequence data (Nesiocypraea midwayensis, N. lisetae and N. from , the single, living rep- aenigma). While sharing a most recent com- resentative from an older, more diverse genus mon ancestor, the three genera are highly di- of European affinities. Previously, the vergent from each other, representing paraphyletic arrangement of the genera significant periods of independent history. Two INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 133

FIG. 3. Basal Compartment cladogram and phylogram. Bootstrap values are presented above branches in the cladogram and rescaled decay values below. Bolded taxa are new additions to the data set. Their identity number shown in parentheses follows the listing in the Appendix. Generic or suprageneric groupings are indicated to the right of the cladogram. OTUs with an asterisk (*) are not ESUs based on molecular criteria. Phylogram to the right is based on likelihood distances using a GTR+I+G model of sequence evolution. Note that the scaling for branch lengths changes between Ovulidae and Cypraeidae. 134 MEYER are known exclusively from the Indo-Pacific lamarckii populations of the western Indian (Nesiocypraea and Ipsa) and one exhibit a recent divergence from the (Propustularia) from the western Atlantic, but previously recorded E. lamarckii cf. redimita of has a fossil record from North America, the the Andaman Sea. One final finding from addi- Caribbean, and Europe (Kay, 1996). The splits tional Erosaria sequence data is that haplotypes among these ancient groups are among the from Erosaria miliaris and E. eburnea individu- earliest of all extant species and may have oc- als interfinger, indicating that either the diver- curred in the Mesozoic. While reasonably sup- gence between these two taxa is very recent ported as a clade, this basal group is not and lineage sorting has not occurred, or that strongly supported as the most basal sister, and these two taxa represent a cline across the in other analyses (3:1) moves up to become western Pacific from a colored dorsum in the sister of the remaining basal taxa (Erosariinae). west to white shells in the east. The final six genera from the basal compart- ment form the strongly supported clade Mid1 Compartment (Fig. 4) Erosariinae and is the sister group to all remain- ing extant species. Membership and relation- The second paraphyletic compartment con- ships within the Erosariinae are consistent with tains mostly large-shelled taxa from the follow- previous findings (Meyer, 2003). Five taxa from ing tribes: Umbiliini, Cypraeini, Mauritiini, Luriini, Erosaria are added: Erosaria marginalis, E. Austrocypraeini, and the genus . All citrina, E. helvola cf. callista, E. macandrewi, six clades are well supported (> 70/> 5) except and E. englerti. Ten independent lineages are for Austrocypraeini. As in Meyer (2003), inter- strongly supported (bootstraps > 90/decays > relationships among these major suprageneric 6) within Erosaria, but interrelationships among clades are resolved in the consensus, but poorly them are not (< 50/< 4). Erosaria marginalis supported. Austrocypraeini and Luriini are sis- and E. citrina, both from the western Indian ters and recognized as the subfamily Luriinae. Ocean, are strongly supported as sister taxa. Barycypraea and Zoila are sisters and recog- This clade is poorly supported as sister to the nized as the subfamily Bernayinae. Cypraeini E. helvola complex. Within Erosaria helvola, and Mauritiini are sisters and recognized as the three ESUs are identifiable: E. helvola subfamily Cypraeinae. In the current topology, hawaiiensis from Hawaii, E. helvola cf. callista Pustularia and all remaining cowries share a from the Marquesas, and E. helvola helvola more recent common ancestor. This large clade from the remainder of the IndoPacific. The is sister to Luriinae, which in turn is sister to newly included ESU, E. helvola cf. callista, may Bernayinae, and this inclusive clade is sister to need a new name, because the type locality of Cypraeinae. As in Meyer (2003), Umbiliini is sis- E. helvola callista is Tahiti (Shaw, 1909), not ter to all remaining mid1, mid2 and derived taxa. the Marquesas. These five taxa are sister to Within the mid1 compartment, 13 taxa are the remaining Erosaria; however, the basal po- added to the sequence database. The first addi- sition is poorly supported. Erosaria turdus is a tion falls within the genus Umbilia and is tenta- monotypic, deeply divergent lineage. Newly tively recognized as Umbilia cf. petilirostris. A added Erosaria irrorata, a species restricted to single divergent sequence was generated from the oceanic islands of the Pacific, is poorly sup- tissue samples collected from the deep waters ported as sister to a strongly supported clade in the Capricorn Channel off Queensland, Aus- (97/12) including E. albuginosa and E. poraria. tralia. Seven sequenced individuals were com- These three taxa are sister to a well-supported pletely identical, while an eighth sample from a lineage (92/6) of eight taxa that I tentatively rec- subadult shell was significantly divergent. This ognize as Paulonaria at the subgeneric level. single sample may represent the newly de- New sequence data from Erosaria macandrewi, scribed Umbilia petilirostris Darragh, 2002; how- a taxon, closely ally that species with ever, authors disagree on its taxonomic status E. beckii. These two species are sister to the (Wilson & Clarkson, in press). Until more com- remaining Paulonaria taxa. The final additional prehensive sampling is done in the region, I taxon within Paulonaria is Erosaria englerti, a present the divergent sequence as a different species endemic to Easter Island and Sala y ESU, which does not preclude it from being Gomez. Erosaria englerti shares a more recent lumped within U. capricornica at a later date common ancestor with the remaining five with more exhaustive sampling. The relation- Paulonaria taxa. All other relationships within ships within Umbilia remain as in previous Erosaria are the same as those presented in analyses (Meyer, 2003). Meyer (2003) and are indicated in Figure 3. The second taxon added to mid1 is Lepori- Newly added haplotypes from E. lamarckii cypraea mappa aliwalensis from Natal, South INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 135

FIG. 4. Mid 1 Compartment cladogram and phylogram. All other information as in Fig. 3. 136 MEYER

Africa, and falls as sister to previous findings, the interrelationships among mappa rosea. Lorenz (2002) has recently re- major lineages in are poorly supported. vised the of the mappa group in light Consensus methods and poor support result of molecular findings. Importantly, the names I in two polytomies (Fig. 4). Further genetic data associated previously with ESUs have will be needed to address this region of the phy- changed, and those changes are reflected in logeny as all extant taxa have been sampled. the Appendix and also discussed herein. The New sequence data from Barycypraea taxon I previously recognized as Leporicypraea teulerei and B. fultoni place them as sister taxa mappa viridis from SE is now recog- and align them with the genus Zoila to form the nized as Leporicypraea admirabilis. The taxon group Bernayinae. Sequence data presented I previously recognized as Leporicypraea for Barycypraea fultoni are of B. fultoni amorimi mappa panerythra from the non-continental from . The Australian Zoila portions of the western Pacific is now recog- marginata complex is split into two ESUs as nized as viridis. The other increased sampling indicates fixed molecular taxon names remain the same. Sequences of differences between populations separated by L. mappa “rewa” from Pacific localities (Fiji, the Southwest Cape region between capes Vanuatu, Palau, and South China Sea) Naturaliste and Leeuwin. Further sampling di- interfinger with haplotypes of L. mappa rectly within this region may uncover interme- geographica individuals from lo- diate haplotypes that would link the two ESUs calities (NW , Phuket, , and and suggest a cline instead of two independent Zanzibar). Therefore, I recognize only a single lineages. Such a finding is the case in the Zoila taxon, L. mappa geographica, for this clade. friendii complex. However, as none have been Because of its conchological distinctiveness discovered yet, I present the data as two tenta- and sympatry with conspecifics, Lorenz (2002) tive ESUs: Zoila marginata marginata to the elevated L. mappa geographica to specific sta- south and Z. marginata ketyana to the west. tus with Indian and Pacific subspecies. Based Other described Z. marginata taxa (Lorenz, on the genetic difference between mappa-com- 2001; 2002) within each ESU interfinger, and plex conspecifics and geographic overlap, spe- do not fulfill molecular criteria for recognition. cific status is certainly acceptable. However, the Sequence data from Zoila mariellae are the fi- remaining L. mappa subspecies are para- nal addition to the Bernayinae clade. While the phyletic. The phylogeny Lorenz (2002: 27) pre- exact provenance of the sequenced is sents is correct and reflects this arrangement. unknown, it is likely from the northwestern shelf Certainly, other recognized cowrie species are of Australia. Molecular results place Z. mariellae derived from paraphyletic parent species (e.g., as a distinct sister to Z. decipiens, also from Eclogavena coxeni and others; see Meyer, the northwestern shelf, as expected. 2003: table 4, and cases herein), and L. Following along the phylogeny, the clade geographica would have to be added to this Luriinae comes next. and are list. These results suggest a third species sis- strongly supported as the clade Luriini. A small ter to L. geographica should be recognized that fragment from 16S was amplified from a de- would include both L. mappa viridis and L. graded Talparia exusta specimen, and as ex- mappa admirabilis. L. mappa geographica in- pected, the taxon is sister to the more dividuals have been found sympatrically with widespread . Surprisingly, se- both L. mappa mappa and L. mappa viridis in- quence divergence between the two species dividuals in the Pacific Ocean. However, as yet, appears to be relative small, indicating a more L. mappa mappa and L. mappa viridis haplo- recent divergence than expected. Better-pre- types have not been found together. served material from T. exusta is needed be- One new undescribed taxon is added to fore these relative results can be confidently Mauritia. Haplotypes of M. arabica individuals assessed. The inclusion of four new taxa to the from American Samoa cluster independently Austrocypraeini (Arestoides argus contrastriata, from haplotypes of M. arabica individuals from Lyncina broderipii, L. ventriculus from the In- other Pacific localities. Shells from Samoan in- dian Ocean, and L. kuroharai) does not help in dividuals tend to be smaller, more heavily mar- resolving interrelationships among member gined and more circular than individuals from taxa. Arestoides argus is broken into a Pacific other Pacific localities. Results from increased clade, A. argus argus, and a western Indian sampling in both M. depressa depressa (N = Ocean clade, A. argus contrastriata, based on 10) and M. depressa dispersa (N = 10) main- additional sequence data from the Indian tain their independent, reciprocally monophyl- Ocean. Lyncina broderipii appears as sister to etic status, albeit recently diverged. As in L. nivosa within the Callistocypraea clade, as INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 137 predicted in Meyer (2003). A single sampled least one ESU is added within each genus ex- individual of L. ventriculus from Christmas Is- cept the monotypic Neobernaya. Pseudo- land in the Indian Ocean falls significantly out- zonaria nigropunctata, a Galapagos endemic, side the haplotype cluster of individuals (N = 6) falls into the eastern Pacific clade as a diver- from various regions of the Pacific basin. gent sister to P. arabicula, although not strongly Lyncina ventriculus is an oceanic taxon, and supported. The position of Schilderia achatidea because of the geographic gap between sites has been mentioned previously as sister to across continental Southeast Asia, I choose to Zonaria, now found exclusively in the eastern present the Christmas Island form as new, Atlantic. Two taxa are added from Zonaria. undescribed, distinct ESU. Further sampling of Zonaria picta from the Cape Verde Islands falls individuals from Christmas Island may change near the base of Zonaria, and its relationship this interpretation, but they are currently lack- with other Zonarid taxa is ambiguous, resulting ing. A single sample of Lyncina kuroharai was in a polytomy at the base of the group. Alterna- sequenced and the results place it closely re- tive phylogenetic reconstructions at the base lated to L. sulcidentata, an endemic Hawaiian of the group show small internodes, indicative taxon. The shallow split between these two taxa of a short radiative burst, with little divergence indicates a relatively recent common ancestor. since. New sequence data from Pseudozonaria Faunal ties have been documented in other angelicae are extremely similar to haplotypes cowrie species between Hawaii and , from P. pyrum (both P. pyrum angolensis and most notably in , and the close P. pyrum senegalensis). I include P. angelicae affinities between L. kuroharai and L. as a taxon in the phylogeny, but prefer to con- sulcidentata represent another example of this sider it at most a subspecies until further se- biogeographic link. quence data are available within the P. pyrum The final two ESUs added within the mid1 complex, as I have reservations concerning di- compartment are members of the genus vergences along the mostly continuous West Pustularia, and more specifically are recognized African/Mediterranean coastline. subspecies of Pustularia bistrinotata. A single Sequence data from six additional taxa are P. bistrinotata keelingensis individual was se- included within Cypraeovulinae, two from quenced, is distinct, and appears as sister to Notocypraea and four from Cypraeovula. In the remaining P. bistrinotata complex. Further- Notocypraea, I tentatively recognize two ESUs more, P. bistrinotata sublaevis individuals (N = within Notocypraea angustata, with a phyloge- 5) from southeast Polynesia (Tuamotu and netic break somewhere between Port Lincoln Societies) cluster together, forming a third ESU and Port Macdonnel, South Australia. Two di- within P. bistrinotata. vergent haplotype clusters exist without inter- mediate states. Again, further data may change Mid2 Compartment (Fig. 5) this interpretation, but at present I chose to rep- resent these as different ESUs indicating dis- The third phylogenetic compartment, mid2, tinct evolutionary trajectories. Sequence data contains members from the genera from a single specimen of Notocypraea Neobernaya, Pseudozonaria, Schilderia, hartsmithi, a rare species from southeastern Zonaria, the subfamily Cypraeovulinae, and the Australia, indicate that the species is sister to tribe Bistolidini of the subfamily Erroneinae. all remaining Notocypraea taxa. Within Interrelationships among member clades are Cypraeovula, four taxa are added, but their in- consistent with previous findings (Meyer, 2003). clusion does not change previous interpreta- Neobernaya and Pseudozonaria are sisters, tions that the group is composed of and that clade is sister to the remaining cow- predominately four divergent lineages with mi- ries. The inclusion of sequence data from the nor differences within each. New sequence data genus Schilderia (S. achatidea), place the from both Cypraeovula fuscorubra and C. group as sister to Zonaria, and together this fuscodentata closely align these taxa with C. clade shares a more recent ancestor with the capensis. New sequence data from C. mikeharti remaining taxa. The subfamily Cypraeovulinae and C. algoensis closely align those taxa with includes both the South African Cypraeovula C. edentula and C. alfredensis. Noting the shal- and South Australian Notocypraea and is sis- low divergences among recognized species in ter to the western IndoPacific Erroneinae, which Figure 5, I am doubtful that many of the de- is composed of two tribes: Bistolidini and scribed subspecies within Cypraeovula (sum- Erroneini. marized in Lorenz, 2002) will fulfill my Within the mid2 compartment, 25 taxa are molecular criteria for ESU status. As some added to the existing sequence database; at species are differentiated currently by only a 138 MEYER

FIG. 5. Mid 2 Compartment cladogram and phylogram. All other information as in Fig. 3. INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 139 single mutation (e.g., Cypraeovula mikeharti/ lineages remain strongly supported (P. asellus, C. algoensis or C. castanea/C. iutsui), there P. clandestina/diluculum, P. ziczac and P. simply is not enough room for differences to contaminata), but confident hypotheses of other have accumulated between taxa. This is not interrelationships require further data. to say that described entities are not indepen- Three taxa are added to : B. stolida dent. Indeed, because Cypraeovula taxa are diagues, B. owenii and an undescribed, dis- direct developers with limited dispersal and tinct eastern Indian Ocean clade of B. ursellus. gene flow, regional differences are expected Individuals of B. stolida diagues from the on small geographic scales, much like the Seychelles fall as sister to B. stolida South Australian endemic clades Umbilia, rubiginosa. Bistolida owenii, a western Indian Zoila, and Notocypraea. However, based on the Ocean taxon, is sister to the Red Sea endemic genetic similarity among sampled member B. erythraeensis. A new Bistolida ursellus se- Cypraeovula, much of this variation has to be quence from the Andaman Sea is poorly sup- very recently derived. This pattern is borne out ported as sister to the remaining B. ursellus in the South Australian direct developers that taxon from the Pacific basin. Its placement is have been more extensively sampled. equally parsimonious as either sister to B. The tribe Bistolidini within Erroneinae is com- ursellus (Pacific) or forming a B. ursellus grade posed of members from five genera: leading to the B. kieneri lineage. The topology , Bistolida, , of the two B. ursellus taxa as sisters is more and Cribrarula. As in Meyer (2003), the basal likely and consistent with morphology. root of Bistolidini is poorly resolved. Overall One taxon is added to Ovatipsa and two taxa analyses place either Palmadusta as sister to to Talostolida. Within Ovatipsa, the subspe- the other four genera or Palmadusta and cies O. chinensis amiges from the Pacific ba- Bistolida as a clade, sister to the remaining sin and Western Australia is distinct from O. three. Compartmentalized analyses place chinensis chinensis from the west- Palmadusta at the base, although poorly sup- ward through the Indian Ocean to the east ported. The addition of 15 ESUs did not help in coast of Africa. Various other O. chinensis resolving this issue. Only one taxon is added subspecies have been described within the to the Palmadusta clade, but it alters the sub- Indian Ocean (e.g., Lorenz & Hubert, 1993), specific designations previously ascribed and preliminary data indicate that these Indian (Meyer, 2003). New haplotypes from Andaman Ocean subspecies may represent very recent Sea P. clandestina individuals form a distinct divergences within what I am currently recog- monophyletic clade. This new ESU is sister to nizing as O. chinensis chinensis. However, the western Indian Ocean P. clandestina until more individuals are sampled, I maintain passerina, and the two of them are sister to the them all under the taxon Pacific P. clandestina clade and the Japanese chinensis. Within Talostolida, two taxa are endemic P. artuffeli. Based on a review of P. added that appear as sisters to each other: T. clandestina subspecies and type localities, the subteres from southeastern Polynesia and T. Pacific clade that I had formerly (Meyer, 2003) latior from Hawaii. These two taxa are sister recognized as P. clandestina clandestina should to Talostolida pellucens. All four taxa currently be P. clandestina candida, and the new P. included within Talostolida are deeply divergent clandestina clade from the Andaman Sea now independent ESUs. A single haplotype of bears the name P. clandestina clandestina. I Talostolida teres “alveolus” (sensu Lorenz, also reviewed the subspecies and type locali- 2002) is completely identical to haplotypes of ties for the three P. asellus ESUs previously T. teres teres individuals from both the Society unnamed (Meyer, 2003). Based on increased Islands and the Tuamotu. Moreover, T. teres sampling and conchological comparisons, I ten- individuals from SE Polynesia have been de- tatively ascribe the following subspecific des- scribed by Lorenz (2002) as a distinct subspe- ignations for the three clades: P. asellus asellus cies T. teres “janae”; however sampled for the western Indian Ocean clade, P. asellus individuals of T. teres from SE Polynesia vespacea for the Seychelles to western Pacific interfinger with individuals sampled from the clade, and P. asellus bitaeniata for the Western Pacific (Papua New Guinea and Melanesian and Pacific clade (Fig. 5, Appen- Guam). Therefore, the data do not support T. dix). Unfortunately, the addition of P. clandestina teres “janae” as a valid taxon, based on my clandestina does not help in resolving the basal criteria. All Marquesan individuals sequenced nodes of Palmadusta. As shown in Figure 5, possess T. pellucens haplotypes, whereas all the base of Palmadusta is poorly resolved and T. teres-like individuals from the remainder of sister group assignments are ambiguous. A few SE Polynesia possess T. teres haplotypes. 140 MEYER

The Cribrarula clade includes eight additional analyses and discussion of this fascinating, taxa, making it the most diverse genus within species-rich group is in preparation (Meyer et Bistolidini. Two taxa, Cribrarula taitae from al., in prep.). American Samoa and C. garciai from Easter Island, are added to the deeply divergent Pa- Derived Compartment (Fig. 6) cific subclade. Both taxa are recently divergent members from their respective sister taxon. The final compartment analyzed is the derived Cribrarula taitae appears as a closely related monophyletic clade recognized as the tribe sister to C. catholicorum, and C. garciai is Erroneini. This clade includes the following nine closely related to C. cumingii. Only a single in- genera: Austrasiatica, Palmulacypraea, dividual from each of the two taxa was included Erronea, Purpuradusta, Contradusta, Nota- in these analyses, and the results would be dusta, Eclogavena, and . better addressed with multiple samples. Two Many (25) taxa are added within the tribe, and members are added to the Western Indian phylogenetic analyses result in some surpris- Ocean subclade: Cribrarula pellisserpentis and ing affinities. For the most part, major genera C. esontropia francescoi, both from Madagas- are well supported, but their interrelationships car. Cribrarula esontropia francescoi is a closely are not. Three taxa currently ascribed to related sister to C. esontropia esontropia, which Austrasiatica were included in previous analy- includes C. esontropia cribellum (Meyer, 2003). ses (Meyer, 2003); however, they were consid- Cribrarula pellisserpentis is a deeply divergent ered as representatives of the genus member within the western Indian Ocean Nesiocypraea. As discussed earlier, the find- subclade and is sister to the other three ESUs. ing that Nesiocypraea teramachii is distantly Four taxa are added to the remaining Cribrarula related raises the subgenus Austrasiatica to member clade. A single individual of C. cribraria generic status for the clade that includes from Masirah, Oman, appears significantly di- Austrasiatica langfordi, A. hirasei and A. vergent from population samples of the previ- sakurai. As in Meyer (2003), Austrasiatica is ously unnamed C. cribraria ESU from the sister to all other Erroneini taxa, followed by Andaman Sea. Conchologically, this individual Pamulacypraea as sister to the remainder. As approximates the western Indian Ocean taxon predicted in Meyer (2003), the newly added C. cribraria abaliena and is tentatively recog- Pamulacypraea musumea falls as sister to P. nized as such. A single individual of C. cribraria katsuae. Even with the addition of 24 taxa (a australiensis from Western Australia falls within 67% increase), the topology among the rest of the Andaman C. cribraria cluster; therefore, I the major Erroneini lineages is ambiguous. Six tentatively adopt the name C. cribraria cf. added “Erronea” species form a basal grade “australiensis” for a taxon that extends from the leading to the Adusta/Erronea split previously Andaman Sea southward to Western Austra- recognized in Meyer (2003). I take a conserva- lia. More exhaustive sampling is required to tive approach and redefine Erronea to include confirm these geographic patterns. A single in- all these taxa and subsume Adusta to a well- dividual of C. exmouthensis magnifica from supported subclade within the group, as the Broome is significantly different from samples new data demonstrate that Adusta and Erronea of C. exmouthensis exmouthensis from the (including the more recent additions) are not Exmouth Gulf region, therefore validating the equivalent (sisters). If Adusta were to be main- status of that taxon. Additional samples of C. tained at equivalent generic status, Erronea cribraria rottnestensis (N = 3) further validate would represent a paraphyletic group. the taxon’s uniqueness. Eight individuals of C. Purpuradusta, Eclogavena, Melicerona and melwardi from northeastern Australia all share Blasicrura are all well-supported monophyletic a common ancestor and are reciprocally mono- lineages. As in Meyer (2003), is well phyletic with respect to the remaining C. supported only if restricted to members of the cribraria individuals. Moreover, a single C. Notadusta punctata complex. However, be- cribraria cribraria individual from the same reef cause Notadusta martini is often considered a (Lamont Reef in the Bunker Group) clusters as member of Notadusta, I include it within expected with other Pacific C. cribraria cribraria Notadusta here, although poorly supported. In individuals. The final taxon included is C. a similarly conservative manner, I include two cribraria abrolhensis (N = 3), and haplotypes of the added taxa within Contradusta, although are shallowly divergent but reciprocally mono- again poorly supported. Support for relation- phyletic with respect to samples of C. cribraria ships among these seven genera is poor and cribraria (N = 30) from predominately western is likely because of the short internode length Pacific localities (Appendix). More thorough between divergent lineages. INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 141

FIG. 6. Derived Compartment cladogram and phylogram. All other information as in Fig. 3. 142 MEYER

Twelve additional taxa are added to Erronea. Andaman Sea (Meyer, 2003). The Erronea Six of the additions are traditionally recognized caurica complex and the associated E. as distinct species, four have been recognized cylindrica, E. ovum and E. errones species will as subspecies, and two are newly discovered, be more thoroughly addressed in another pa- but may have names associated with them that per (Meyer, in prep.) as the group exhibits re- have been placed into synonymy. Of the new markable geographic structuring, polyphyly of species, three form a relatively well-supported recognized species (E. ovum), and evidence clade: Erronea rabaulensis shares a more re- of introgression based on nuclear markers. cent common ancestor with E. fernandoi (80/1), Purpuradusta is well supported and contains and those two are sister to E. vredenburgi (84/ four newly added taxa that fall in expected re- 3). The three additional Erronea species all nest lationships. The southeastern Polynesian en- deeply within the clade, and their relationships demic species Purpuradusta oryzaeformis is are not well supported. Erronea pallida appears distinct and sister to P. minoridens that ranges as sister to the clade of the previously described throughout the remainder of the western three species and Adusta. Erronea pyriformis IndoPacific. A single specimen of P. microdon is relatively well supported (81/6) as the sister from falls outside the haplotype to the clade previously recognized as Erronea clade of other sampled individuals from the (Meyer, 2003). Finally, Erronea xanthodon falls Pacific basin (N = 5). This East African popula- at the base of Erronea and is sister to all other tion is recognized as Purpuradusta microdon Erronea taxa. Within the crown Erronea chrysalis. Two peripheral populations of subclade, six taxa are added that are all tradi- Purpuradusta fimbriata in the Pacific Basin are tionally recognized at the subspecific level. In- introduced. First, Hawaiian populations of P. dividuals of Erronea cylindrica lenella (N = 8, fimbriata are distinct (N = 7) and were previ- all from New Caledonia) form a monophyletic ously recognized as P. fimbriata waikikiensis; group strongly supported (91/6) as sister to the thus this name is resurrected as a valid entity. clade including the remaining E. cylindrica in- Second, individuals from the Marquesas are dividuals plus two subspecies of E. ovum. also distinct genetically, consistent with the These results imply that E. cylindrica at the subspecies designation of Lorenz (2002), P. specific level is a paraphyletic taxon. Newly fimbriata marquesana (N = 14). Both of these added individuals of Erronea ovum ovum from Pacific P. fimbriata subclades share a more both Singapore and the Philippines (N = 15) recent history with the widespread Pacific sub- form a monophyletic group sister to E. ovum species P. fimbriata unifasciata, as expected. palauensis (N = 7). The four remaining, newly Two newly added species, “Erronea” barclayi added taxa are all members of the Erronea and “Erronea” pulchella, come out as sister caurica complex. First, individuals (N = 7) of species in phylogenetic analyses. Moreover, the newly described E. caurica samoensis ap- these two taxa appear as sister to Contradusta pear as a distinct lineage sister to individuals in the most likely topology. Because of these (N = 15) from the remainder of the Pacific and results, and the poorly supported nature of their Western Australia (E. caurica caurica). Four relationships, I tentatively place the two taxa in geographically structured haplotype clades are the genus Contradusta, with the caveat that found exclusively in the Western Indian Ocean. they may be removed with future data. These Erronea caurica dracaena is currently restricted results are somewhat surprising, particularly to the Seychelles based on sampling. Newly because “Contradusta” pulchella is thought to added individuals from East Africa and Mada- be closely related to Erronea pyriformis be- gascar form a haplotype clade that I recognize cause of the darkly stained columellar denti- as Erronea caurica elongata. Individuals of E. tion and overall conchological similarities. The caurica quinquefasciata from the Red Sea, East sister relationship between Contradusta Africa and Oman form the third monophyletic pulchella and C. barclayi is more acceptable group. Finally, newly sequenced individuals as their divergence is deep, and the phyloge- from Masirah (N = 7) form a private haplotype netic affiliations of C. barclayi were more diffi- clade (E. caurica ssp. #1) sister to E. caurica cult to predict based on morphological criteria. quinquefasciata. The final, newly added taxon Another surprising result is the sister relation- (E. caurica ssp. #2) within the E. caurica com- ship between Notadusta martini and “Erronea” plex is a clade (N = 18) that includes individu- hungerfordi. Given these phylogenetic results, als primarily from India, but with a few I tentatively place “Erronea” hungerfordi within individuals from Masirah, Oman. This haplo- Notadusta, but with little confidence, although type clade is sister to the clade recognized pre- it is reasonably supported (73/4), and suspect viously as E. caurica cf. derosa from the that it may be removed with more samples and INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 143 sequence data. Within the remaining Notadusta and I will use his checklist (pp. 250−291) as a complex, individuals of N. punctata trizonata benchmark for comparisons. Lorenz recog- (N = 9) form a monophyletic group sister to the nizes 232 species, of which I have sequenced Pacific N. punctata punctata clade. Finally, in 210 (> 90%), and they are presented herein. regards to Notadusta, “Notadusta” rabaulensis The missing species are as follows: was mentioned previously as a member of Nesiocypraea aenigma, N. lisetae, N. midway- Erronea and “Notadusta” musumea as ensis, Austrasiatica alexhuberti, Erosaria Palmulacypraea, further reducing the member- ostergaardi, Zoila perlae, Lyncina camelopar- ship of Notadusta (Meyer, 2003). dis, L. joycae, Pustularia chiapponii, Cypra- The final four additions to the dataset fall into eovula colligata, C. cruickshanki, C. immelmani, Melicerona and Blasicrura. First, two taxa are Palmadusta androyensis, P. johnsonorum, added to Melicerona. Samples of Melicerona Austrasiatica deforgesi, Palmulacypraea listeri melvilli (N = 5) from Queensland, Austra- boucheti, P. omii, Eclogavena luchuana, lia, form a monophyletic group sister to the re- Erronea (?) angioyorum, and E. nymphae. Se- maining Melicerona taxa. (Two rostrate and quences from samples of both Purpuradusta melanistic individuals interfinger among the barbieri and “Talostolida” rashleighana have other three haplotypes indicating that the tera- been obtained, but were too late for inclusion in tology is likely driven by phenotypic responses these analyses. All missing species are rare, to environmental conditions rather than having with small ranges located generally at the pe- a genetic basis.) Samples of Melicerona felina riphery of their putative sister species based on from both Oman and East Africa form a mono- conchological and anatomical characters. Of phyletic group, and because the haplotypes the 210 sequenced species, phylogenetic com- from the two regions interfinger, there is no parisons and molecular criteria support all but evidence for a distinction between the subspe- 15 (93%) as ESUs. The 15 recognized species cies M. felina felina and M. felina fabula. Within not supported by my criteria are discussed be- Blasicrura, two taxa are added, based on the low. For Nucleolaria granulata, Monetaria sequencing results. First, samples of Blasicrura obvelata, Erosaria eburnea, Zoila orientalis, Z. summersi, a Fijian and Tonga endemic, appear thersites, Luria controversa, L. gilvella, as a recently divergent sister to the also newly Notocypraea occidentalis, and Palmadusta included B. pallidula cf. vivia from American humphreysii, multiple individuals were se- Samoa. This clade is sister to the Melanesian quenced and the haplotypes interfingered subspecies Blasicrura pallidula rhinoceros, as within their closest relative. For the next six expected based on geography. This resulting species that I do not support, only a single in- topology indicates that the Blasicrura pallidula dividual was sequenced, thus they may indeed complex is paraphyletic. represent a very young independent trajectory. However, when compared to the genetic diver- sity within their closest relative, the genetic dif- DISCUSSION ference is unremarkable, and in some instances, only a single mutation different from The ultimate goal of this project is to construct putative conspecifics: Zonaria angelicae, Z. a comprehensive phylogeny of cypraeid gas- petitiana, Cypraeovula mikeharti, Bistolida tropods at the appropriate level for diversifica- brevidentata, Cribrarula garciai, and C. taitae. tion studies. From a molecular perspective, all While genetic data are overall broadly con- ESUs presented are effectively equal units of sistent with taxa recognized at the specific level, diversity, whether they are currently recognized the results are even more remarkable when as species, subspecies or some other level. compared among taxa recognized at subspe- There are some noted exceptions as OTUs cific levels. Lorenz recognizes 260 taxa at the were used on occasion that represented un- subspecific level. Of those 260 subspecies, I sorted or clinal variation within an ESU (e.g., have sequenced at least two individuals from Erosaria miliaris/eburnea). However, on a gen- 160 in order to assess their validity. Molecular eral scale, each taxon shown in the phylogenies criteria support 113 (> 70%) of these taxa as (Figs. 3−6) represents an independent evolu- legitimate ESUs. Moreover, sequence results tionary trajectory. indicate an additional 20 distinct ESUs not Because so much taxonomic information is recognized as subspecies by Lorenz (but available for cowries, it is informative to see sometimes mentioned as important varieties how molecular criteria compare with recog- or forms). A full listing of sampled taxa and nized taxonomic entities. The most recent com- their current ESU status as indicated by the pilation of the cowries is that of Lorenz (2002), prior criteria can be found at the Cowrie Ge- 144 MEYER netic Database Project Website (http:// Mtumwa Mwadini, Peter Ng, Steve Norby, www.flmnh.ufl.edu/cowries). The website in- Shuichi Ohashi, Yoshihiro Omi, Ina Park, Marcel cludes other information, such as localities Pin, Cory Pittman, Xavier Pochon, Matt Rich- sampled, numbers of individuals for each taxon, mond, Raphael Ritson-Williams, Gonçalo Rosa, and photographs of the specimens sequenced. Gary Rosenberg, Teina Rongo, Fred Schroeder, Overwhelming molecular support for tradition- Mike Severns, Pauline Severns, Hung-Long Shi, ally recognized taxa, both at specific and sub- Brian Simison, Michael Small, John Starmer, specific levels, is extremely encouraging. First, Steve Tettlebach, David Touitou, Martin Wallace, from a taxonomic standpoint, these molecular Chia-Hsiang Wang, Dave Watts, Barry Wilson, results corroborate the excellent work done by Woody Woodman, Shu-Ho Wu. The following centuries of malacological researchers, at both institutions are acknowledged: Florida Museum professional and amateur levels. Similar mo- of Natural History; University of California Mu- lecular surveys of other diverse groups will pro- seum of Paleontology; Academy of Natural Sci- vide valuable comparisons in order to assess ences of Philadelphia; Bernice P. Bishop taxonomic congruence (e.g., Jackson & Museum, Honolulu, Hawaii; California Academy Cheetham, 1990) and address concordant di- of Sciences; Institute of Marine Sciences, Zan- versification patterns. Second, from a molecu- zibar; University of Dar es Salaam; Jackson- lar perspective, sequence data provide a ville Shell Club; Musée National d’Histoire suitable, objective, relative metric for circum- Naturelle, Paris, France; National Museum of scribing appropriate evolutionary units. Assum- Natural History Naturalis, Leiden, The Nether- ing rate constancy in the molecules (COI only, lands; Santa Barbara Museum of Natural His- in prep.), molecular divergences can constrain tory; National Museum of Natural History; and the tempo of diversification and assess the dis- Suganthi Devadason Marine Research Institute. tinctiveness of purported taxa. A growing body I also would like to thank Felix Lorenz, Jr., for of molecular data across the diversity of life his thoughtful comments, as well as the reviews undoubtedly will provide insight to some of our of four anonymous reviewers. Final decisions most fundamental evolutionary questions. and opinions are wholly mine. This research has been financially supported by the following NSF grants: DEB-9807316, ACKNOWLEDGEMENTS DEB 0196049, and OCE-0221382.

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dase subunit I from diverse metazoan inver- PALUMBI, S. R., 1996, Nucleic acids II: The tebrates. Molecular Marine Biology and Bio- polymerase chain reaction. Pp. 205−247, in: technology, 3: 294−299. D. M. HILLIS, C. M. MORITZ & N. K. MABLE, eds., GROVES, L. T., 1994, Catalog of fossil and Re- Molecular systematics, 2nd ed., Sunderland, cent Cypraeidae and Eocypraeinae (Ovu- Massachusetts, USA: Sinauer Associates, lidae) described since 1971. The Cowry, (n.s.) Inc. 1: 5−16. POSADA, D. & K. A. CRANDALL, 1998, Model- JACKSON, J. B. & A. H. CHEETHAM, 1990, test: testing the model of DNA substitution. Evolutionary significance of morphospecies: a Bioinformatics, 14: 817−818. test with cheliostome Bryozoa. Science, 248: RIESE, K., 1931, Phylogenetische Betrachtung 579−583. über das Nervensystem von Cypraea moneta KAY, E. A., 1957, The systematics of the auf Grund seiner Morphologie und Histologie. Cypraeidae as elucidated by a study of Jenaische Zeitschrift für Naturwissenschaft, Cypraea caputserpentis and related forms. 65: 361−486. Zoology. Hawaii, University of Hawaii. RISBEC, J., 1937, Anatomie des Cypraeidae. KAY, E. A., 1960, Generic revision of the Archives du Muséum National d’Histoire Cypraeinae. Proceedings of the Malacological Naturelle, Paris, 14: 75−104. Society of London, 33: 278−287. SCHILDER, F. A., 1936, Anatomical characters KAY, E. A., 1963, Anatomical notes on Cypraea of the Cypraeacea which confirm conchological aurantium Gmelin and other cowries and an classification. Proceedings of the Malacologi- examination of the subgenus Lyncina cal Society of London, 22: 75−112. Troschel. Journal of the Malacological Society SCHILDER, F. A., 1939, Die Genera der of Australia, 7: 47−61. Cypraeacea. Archiv für Molluskenkunde, 71: KAY, E. A., 1984, Patterns of speciation in the 165−201. Indo-West Pacific. Pp. 15−31, in: F. J. SCHILDER, F. A., 1965, The geographic distribu- RADOVSKY, P. H. RAVEN & S. H. SOHMER, eds., tion of cowries (Mollusca: Gastropoda). The Biogeography of the tropical Pacific. Bishop Veliger, 7: 171−183. Museum Special Publication No. 72: Honolulu. SCHILDER, F. A., 1969, Zoogeographical stud- KAY, E. A., 1985, About the cowries. Pp. 4−14, ies on living cowries. The Veliger, 11: 367−377. in: C. M. BURGESS, ed. Cowries of the World. SCHILDER, F. A. & M. SCHILDER, 1938, Pro- Cape Town, Republic of South Africa: G. drome of a monograph on living Cypraeidae. Verhoef Seacomber Publications. Proceedings of the Malacological Society of KAY, E. A., 1990, Cypraeidae of the Indo-Pacific London, 22−23: 119−231. − Cenozoic fossil history and biogeography. SCHILDER, M. & F. A. SCHILDER, 1971, A cata- Bulletin of Marine Science, 47: 23−34. logue of living and fossil cowries. Taxonomy KAY, E. A., 1996, Evolutionary radiations in the and bibliography of Triviacea and Cypraeacea, Cypraeidae. Pp. 211−220, in: J. TAYLOR, ed., Gastropoda Prosobranchia. Memoires du Origin and evolutionary radiation of Mollusca. Institut Royal des Sciences Naturelles de Oxford: Oxford University Press. xiv + 392 pp. Belgique, (2)85: 170−246. LILTVED, W. R., 1989, Cowries and their rela- SHAW, H. O. N., 1909, Notes on the genera tives of southern Africa: a study of the south- Cypraea and Trivia. Proceedings of the Mala- ern African Cypraecean and Velutinacean cological Society of London, 8: 288−313. gastropod fauna. Cape Town, Republic of SORENSON, M. D., 1999, TreeRot. 2.0 ed. Bos- South Africa: G. Verhoef, Seacomber Publica- ton, Massachusetts: Boston University. tions. 208 pp. SWOFFORD, D. L., 1998, PAUP*: Phylogenetic LORENZ, F., 1989, Annotated descriptions of Analysis Using Parsimony (*and Other Meth- some new and old members of Cypraeidae. ods). Sinauer, Sunderland, Massachusetts. Schrifen zur Malakozoologie (Cismar), 2: 138. TROSCHEL, F. H., 1863, Das Gebiss der LORENZ, F., 2000, Beiträge zur Kenntnis der Schnecken zur Begründung einer natürlichen Ringkauri Erosaria annulus (Linné 1758) und Classification. Berlin: Nicolaische Verlags- verwandter Arten (Mollusca: Gastropoda: buchhandlung. Cypraeidae), Schriften zur Malakozoologie, 14: VAYSSIÈRE, A., 1923, Rechereches zoologiques 1−95, 11 plts. et anatomiques sur les mollusques de la famille LORENZ, F., 2001, Monograph of the living des Cypraeides, 1re partie. Annales du Musee Zoila, Hackenheim, Germany: ConchBooks. d’Histoire Naturelle de Marseille, Zoologie 18: 187 pp. 1−120. LORENZ, F., 2002, New worldwide cowries. VAYSSIÈRE, A., 1927, Rechereches zoo- Hackenheim, Germany: ConchBooks. 292 pp. logiques et anatomiques sur les mollusques de LORENZ, F. & A. HUBERT, 1993, A guide to la famille des Cypraeides, 2me partie. Annales worldwide cowries. Wiesbaden, Germany: C. du Musee d’Histoire Naturelle de Marseille, Hemmen. 571 pp. Zoologie 21: 133−184, pls. 24−28. MEYER, C. P., 2003, Molecular systematics of WILSON, B. & P. CLARKSON, in press, Australia’s cowries (Gastropoda: Cypraeidae) and diver- spectacular cowries, a review and field study of sification patterns in the tropics. Biological two endemic genera: Zoila and Umbilia, Odys- Journal of the Linnean Society, 79: 401−459. sey Publishing. MORITZ, C., 1994, Defining ‘Evolutionarily Sig- nificant Units’ for conservation. Trends in Ecol- ogy and Evolution, 9: 373−375. Revised ms. accepted 19 January 2004 146 MEYER FIG. 7. Sampled localities indicated for listed taxa in Appendix (column 3). 7. Sampled localities indicated for listed taxa in FIG. INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 147 ) continues ( bolded. Crossouts

AY161384 AY161617 OUTGROUPS

59 57, Lightfoot, 1786 Sowerby 1832 I, Petuch, 1979 Azuma, 1989 Sowerby 1828 I, Cate, 1964 79 5 Cate, 1973 Cate, 1973 23 Linnaeus, 1758 Cate, 1973 23 25 Adams & Reeve, 1848 1973 & Azuma, Cate Weinkauff, 1881 23 Kuroda, 1958 57 26 22 Linnaeus, 1758 3 25 1810Lamarck, Linnaeus, 1758 25 Linnaeus, 1758 57 Sowerby 1832 I, 25 30 AY161402 59 AY161403 AY161635 28 AY161636 83 AY534351 78 AY534428 AY161391 AY161390 AY161624 AY161623 AY161392 AY534353 AY161393 AY534352 AY161625 AY534430 AY161394 AY161626 AY534429 AY534354 AY161627 AY534431 AY161395 AY534355 AY161628 AY534432 AY161396 AY161397 AY161629 AY161398 AY161630 AY161399 AY161631 AY161400 AY161632 AY161401 AY161633 AY161634 APPENDIX 1: Sampled Taxa and Supplementary Data and Supplementary APPENDIX 1: Sampled Taxa Accession # Accession OTUs Authors Sampled localities* 16S COI 16S localities* Sampled OTUs Authors Pease, 1865 Pease, New taxa and sequence data are numbered and bolded. Additional, newly sampled localities for taxa previously reported are also New taxa and sequence data are numbered bolded. are names previously used in Meyer (2003) that corrected herein because of taxonomic revisions. *for the legend see Fig. 7.

1. Pseudocypraeaexquisita Pedicularia pacifica tokuoi brevis Serratovolva dondani Phenacovolva tokioi rosewateri Crenavolva cf 2. Phenacovolva weaveri 3. Primovulaconcinna 4. Dentiovulatakeoi semperi Prosimnia 5. florida Adamantia verrucosus lacteus Calpurnus ovum Cyphoma gibbosum Simnia aequalis Jenneria pustulata Pseudocypraea adamsonii

148 MEYER ) continues (

AY161407 AY161640 AY161425 AY161658 AY161408 AY161641 AY161408 91

INGROUPS 59 23, 31 5, 8, 30, 71, 5, Perry, 1811 2002 Lorenz, Gray, 1825 Schilder, 1922 Linnaeus, 1758 Pease, 1862 1828 Wood, 1810Lamarck, 89 93 Dautzenberg, 1932 Linnaeus, 1758 59 59, 64 Mighels, 1845 5, 53, 64, 69 Linnaeus, 1758 1810Lamarck, 73 8 5, Linnaeus, 1758 23, 30, 51 5, 9 20, 51, 52, 53, 59 73 5, 8, 10, 30, 52,68 3, 68, 69, 70 AY161417 AY534356 AY161650 AY534357 AY161404 AY534433 AY534434 AY161415 AY161411 AY161637 AY161405 AY161413 AY161648 AY161644 AY161638 AY161646 AY161414 AY161418 AY161412 AY161647 AY161651 AY161645 AY161406 AY161639 AY161416 AY161649 Schilder, 1927 Linnaeus, 1758 Melvill, 1888 Gmelin, 1791 Dillwyn, 1827 1825 Gray, Linnaeus, 1758 73 Melvill, 1888 5, 19, 59, 66, 71, 1909 Shaw, 1810Lamarck, 75 25 1828 Gray, 5, 108 Gray, 1825 Linnaeus, 1758 5, 10, 30, 53, 59, 69, 70 101 100, 1836 Gaskoin, 1870 II, Sowerby 73 Summers & Burgess, 1965 2 71 1, 75 70 69, 19, 59, 69 79 1 59, AY161429 AY161662 AY161409 AY161642 AY534358 AY161410 AY161419 AY534359 AY534435 AY161643 AY161652 AY534436 AY161430 AY161420 AY161428 AY534360 AY161663 AY534361 AY161653 AY534363 AY534437 AY161661 AY534438 AY534440 AY161427 AY534362 AY161660 AY534439 Accession # Accession OTUs Authors Sampled localities* 16S COI 16S localities* Sampled OTUs Authors ) continued (

6. Propustulariasurinamensis 7. Nesiocypraea teramachii neocaledonica Ipsa childreni Cryptocypraea dillwyni Nucleolaria nucleus Nucleolaria granulata Staphylaea limacinainterstincta Staphylaea limacinalimacina laevigata Staphylaea staphylaea Staphylaea semiplota Monetaria obvelata caputophidii Monetaria caputserpentis Monetaria caputdraconis Perisserosa guttata marginalis Erosaria 8. citrina Erosaria 9. Erosaria helvola Erosaria helvola hawaiiensis 10. Erosaria helvola cf. callista turdus Erosaria 11. Erosaria irrorata Erosaria albuginosa Erosaria poraria beckii Erosaria 12. Erosaria macandrewi 13. Erosaria englerti

INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 149 ) continues ( # COI Accession 16S AY161423 AY161656 AY161433 AY161666 AY161436 AY161669 AY161486 AY161719 AY161487 AY161720 AY161481 AY161714 AY161482 AY161715 AY161437 AY161670 AY161422 AY161655 AY161432 AY161665 AY161438 AY161671 AY161478 AY161711 AY161480 AY161713 AY161426 AY161659 AY161435 AY161668 AY161494 AY161727 AY161484 AY161717 AY161485 AY161718 AY161424 AY161657 AY161483 AY161716 AY161477 AY161710 AY161439 AY161672 AY161431 AY161664 AY161434 AY161667 AY161492 AY161725 AY161493 AY161726 AY161421 AY161654 AY161440 AY161673 AY161479 AY161712 AY534364 AY534441 AY534365 AY534442 AY534366 AY534443 localities* localities* 54, 56, 57, 98 98 73 25 55 82 83 52, 82, 22, 23, 5, 10, 14, 32, 14 1, 2 19, 30, 59, 69 5, 10 5, 14 83 30, 59, 71 15, 33 71 5 8 43, 44, 45 38, 50 54, 50 25 84, 85, 86 20, 23 2, 12 2, 48, 49 79 5, 8, 10, 14 5, 83, 101 ; Lorenz,; 2002 71 ; Kenyon,; 1902 55, 57, 59, 64 1888 Gmelin, 1791 Gmelin, 1791 Melvill, Rehder & Wilson, 1975 Rehder & Wilson, 74 Kenyon, 1902 Crosse, 1865Crosse, 1870Sowerby II, Linnaeus, 1758 Gmelin, 1791 Gaskoin, 1849 1849 Gaskoin, Kiener, 1843 Linnaeus, 1758 1817Dillwyn, Melvill, 1888 Gray, 1825 Barnes, 1824 Gmelin, 1791 Linnaeus, 1758 Melvill, 1888 1888Melvill, Verco, 1912Verco, Iredale, 1916 Lorenz, 1989 Darragh, 2002 Kiener, 1843 Linnaeus, 1771 Linnaeus, 1758 Perry, 1811Perry, Gray, 1824 Linnaeus, 1758 Lorenz, 2002 Schilder & Schilder, 1933 1933 Schilder, & Schilder viridis viridis

admirabilis admirabilis

OTUs Authors Sampled OTUs Authors (Indian) cf. chlorizans cf. (Pacific) ) continued ( Leporicypraea mappa viridis mappa Leporicypraea indica Mauritia scurra scurra Mauritia scurra Erosaria kingae Erosaria cernica cernica Erosaria Erosaria thomasi thomasi Erosaria Erosaria spurca spurca Erosaria Erosaria acicularis Erosaria labrolineata Erosaria boivinii Erosaria boivinii Erosaria ocellata gangranosa Erosaria Erosaria lamarckii cf redimitaErosaria lamarckiicf 14. Erosaria lamarckii Erosaria eburnea Erosaria miliaris miliaris Erosaria Erosaria erosa Errosaria nebrites Errosaria nebrites Erosaria erosa Umbilia armeniaca Umbilia hesitata hesitata Umbilia Umbilia capricornica petilirostris 15.Umbilia cf. cervinetta Macrocypraea cervus Leporicypraea valentia rosea mappa Leporicypraea mappa mappa Leporicypraea 16. Leporicypraea mappa aliwalensis Leporicypraea geographica Leporicypraea mappa panerythra mappa Leporicypraea

150 MEYER ) continues ( AY161488 AY161721 AY161465 AY161698 AY534370 AY534447

71 AY161472 AY161705 71 AY161472 69, 70 38, 39, 40 AY161473 AY161706 59, 60, 61, 62, 63, 64, 69, 70, 73 55, 56, 59, 61, 63, 64 Lorenz, 2002 Schilder, 1932Schilder, Lorenz, 2002 59, 66, 73 AY161471 AY161704 Cazenavette, 1846 1846 Cazenavette, Sowerby 1903 III, 1849Gaskoin, Raybaudi, 1978 Cotton, 1948 3 1846Sowerby II, Raybaudi, 1991 106 Smith, 1880 Raybaudi, 1983 43, 44, 45, 46 Gaskoin, 1849 Raybaudi, 1990 39, 40, 41, 43 Cate, 1968 37, 38, 39, 40, 41,42,43 1831Gray, 35 Linnaeus, 1758 32(?) Linnaeus, 1758 33, 34 45, 46 37 AY161497 36 81 AY161730 AY161495 39, 40, 41, 42, 43 10,145, 8, 9, 3, AY534368 AY161728 AY534369 AY534445 AY161496 AY534446 AY161729 15, 18, 19, 23, 30,32,33,52, 56, 57, AY161499 AY534371 AY161498 AY161732 AY534448 AY161502 AY161503 AY161731 AY161489 AY161735 AY161736 AY161500 AY161722 AY161733 AY161501 AY161491 AY161734 AY161724 Lightfoot, 1786 1 AY161490 AY161723 Linnaeus, 1758 5, 19, 52, 70 AY161474 AY161707 5, 7, 8 AY161468 AY161701 8 AY161468 7, AY534444 Schilder & Schilder, 1939 1824Gray, Schilder, 1930 10, 19 1833Dulcos, Gmelin, 1791 Linnaeus, 1758 67 Schilder & Schilder, 1939 2, 3 59, 64, 69 5, 32, 52, 59, 64 2, 11, 13, 15, 19, AY534367 23, 29, 30 9, 19 5, 19, 21, 22, 23, 25,29,30,32, 33, 52, AY161466 AY161475 AY161699 AY161708 AY161469 AY161476 AY161702 AY161709 AY161467 AY161470 AY161700 AY161703

martybealsi maculifera scindata ) (Am. Samoa) Accession # Accession

OTUs Authors Sampled localities* 16S COI 16S localities* Sampled OTUs Authors ( N. Pacific) (Polynesia (Marquesas) (Indian) (Pacific) ) continued ( Mauritia maculifera 18. Barycypraea teulerei teulerei Barycypraea 18. 19. Barycypraea fultoni Zoila marginatamarginata 20. Zoila marginata ketyana Zoila rosselli Zoila venusta Zoila eludens Zoila decipiens 21. Zoila mariellae Zoila thersites Zoila jeanianasherlyae Zoila jeanianajeaniana Zoilafriendii friendii mus tigris Cypraea Mauritia depressa Mauritia grayana Mauritia histrio 17.arabica Mauritia arabica Mauritia arabica asiatica Mauritia arabica immanis Mauritia maculifera Mauritia depressa dispersa

INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 151 ) continues ( # COI AY534452 Accession 16S AY161461 AY161694 AY161458 AY161691 AY161447 AY161680 AY161456 AY161689 AY161449 AY161682 AY161463 AY161696 AY161448 AY161681 AY161460 AY161693 AY161450 AY161683 AY161441 AY161674 AY161507 AY161740 AY161459 AY161692 AY161444 AY161677 AY161508 AY161741 AY161445 AY161678 AY161452 AY161685 AY161446 AY161679 AY161443 AY161676 AY161457 AY161690 AY161455 AY161688 AY161464 AY161697 AY161462 AY161695 AY161442 AY161675 AY161451 AY161684 AY161453 AY161686 AY534373 AY534450 AY534372 AY534372 AY534449 AY534374 AY534451 AY534376 AY534453 98 98 localities* localities* 5 82, 105 57 54 83, 9, 30, 69, 59, 2 97 10 73 73 59, 69, 70 59, 64, 70 23, 76, 79 84, 86, 87 41, 45 5, 8, 9, 20, 30, 65,69,73, 5, 30, 59 87 5, 30, 64 69, 70, 73 8, 30 5, 5, 59, 64 1, 9, 13, 20,30, 1, 52, 53, 55, 59, 69 59 AY161454 AY161687 4 4 13, 14 101 69, 71 Linnaeus, 1758 Sowerby I, 1832 1832 I, Sowerby Linnaeus, 1758 Gmelin, 1791 Gray, 1824 Swainson, 1822 Broderip, 1827 Broderip, 1827 Linnaeus, 1758 Perry, 1811 Sowerby1832 I, Stearns, 1893 1927Schilder, Cate, 1966 Linnaeus, 1758 Linnaeus, 1758 Sowerby 1832 I, Linnaeus, 1758 Gmelin, 1791 Kuroda & Habe, 1961 Iredale, 1939 Linnaeus, 1758 20 Schilder & Schilder, 1938 5, 10 Gray, 1824 Linnaeus, 1758 Schilder & Schilder, 1937 Linnaeus, 1758 1879Garrett, 5, 20, 55, 59, 69, 73 Linnaeus, 1758 Lamarck, 1810Lamarck, 19 AY534375

OTUs Authors Sampled OTUs Authors ) continued ( Talparia talpa 22. Talparia exusta Luria pulchra Luria tessellata Lyncina (Callistocypraea) nivosa Lyncina (Callistocypraea) leucodon Luria isabella 23. Lynicna (Arestoides) argus contrastriata broderipii (Callistocypraea) Lyncina 24. Luria isabellamexicana mariae porteri Miolyncina) Lyncina(cf. argus Lyncina(Arestoides) Trona stercoraria stercoraria Trona Lyncina (Austrocypraea) reevei Chelycypraea testudinaria Lyncina (Callistocypraea) aurantium 26. Lyncina kuroharai schilderorum Lyncina brevirostris Lyncina sulcidentata lynx Lyncina Lyncina leviathan carneola Lyncina propinqua Lyncina Pustularia globulus globulus Lyncina ventriculus ventriculus Lyncina 25. Lyncina ventriculus (Xmas IO)

152 MEYER ) continues ( Schilder & Schilder, 1937Schilder, Schilder & 27, 59, 64 AY161504 AY161737 Schilder & Schilder, 1940 Schilder & Schilder, 1938 98 Burgess, 1967 69, 70 Linnaeus, 1758 1817Dillwyn, Swainson, 1823 Dall, 1909 1810Lamarck, Gray, 1828 73 Hildalgo, 1906 5, 14, 59, 64, 69 1837 I, Sowerby 59 77 Gmelin, 1791 Gray, 1824 Gmelin, 1791 79 Schilder, 1928 78 Odhner, 1923 84 80 90 Clover, 1974 AY534378 Schilder, 1967 AY534377 87 1847Gray, AY534455 AY534454 Reeve, 1846 AY161506 86 87 1847Gray, 87 AY161739 Gmelin, 1791 88 1870 87Sowerby II, 96 Liltved, 1983 AY161505 Higgins, 1868 AY161738 41, 43, 45, 48 Shikama, 1974 41 AY161509 AY161517 Gray, 1828 AY161742 AY161750 43, 45, 47, 48, 49 47, 48 Shaw, 1909 47, 48 AY161519 Gray, 1825 AY161518 Schilder, 1930 AY161752 AY534380 AY161751 AY161520 102 AY534379 6 AY534457 102 AY161753 AY534456 AY161512 AY161745 102 AY534381 6 AY161513 AY161510 AY534458 102 AY161746 6 AY161524 AY161511 AY161743 AY161521 AY161757 AY534382 AY161744 AY534383 AY161754 AY534459 AY534460 AY161525 AY161522 AY161523 AY161758 AY161755 AY161756 AY161527 AY161528 AY161760 AY161526 AY161761 AY161759 AY161530 AY161763 AY534385 AY161529 AY534462 AY534386 AY161762 AY534463 (Port Lincoln) (Port 45 AY534384 AY534461 Accession # Accession OTUs Authors Sampled localities* 16S COI 16S localities* Sampled OTUs Authors ) continued ( Pustularia bistrinotata 28. Pustularia bistrinotata sublaevis mauiensis Pustularia Pustularia margarita Neobernaya spadicea Pseudozonaria annettae Pseudozonaria arabicula 29. Pseudozonaria nigropunctata robertsi Pseudozonaria 30. Schilderia achatidea sanguinolenta Zonaria 31. Zonaria picta Zonaria zonaria senegalensis pyrum Zonaria Zonaria pyrum angolensis 32. Zonaria pyrum angelicae 33. Notocypraea hartsmithi Notocypraea piperita Notocypraea pulicaria Notocypraea comptoni Notocypraea angustata 34. Notocypraea angustata Cypraeovula connelli Cypraeovula castanea Cypraeovula iutsui Cypraeovula capensis fuscorubra Cypraeovula 35. fuscodentata Cypraeovula 36. Cypraeovula coronata 27. Pustularia bistrinotata keelingensis Notocypraea declivis

INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 153 ) continues ( # COI AY534466 Accession 16S AY161539 AY161773 AY161547 AY161781 AY161536 AY161770 AY161548 AY161782 AY161550 AY161784 AY161534 AY161768 AY161535 AY161769 AY161531 AY161764 AY161541 AY161775 AY161553 AY161787 AY161546 AY161780 AY161555 AY161789 AY161549 AY161783 AY161533 AY161767 AY161544 AY161778 AY161558 AY161792 AY161545 AY161779 AY161532 AY161765 AY161557 AY161791 AY161554 AY161788 AY161537 AY161771 AY161543 AY161777 AY161552 AY161786 AY161556 AY161790 AY534388 AY534465 AY534387 AY534464 AY534390 AY534467 AY534392 AY534469 AY534392 AY534391 AY534468 28 12 12 localities* localities* 23, 51, 64 8 51 14, 15, 52, 53, 55,59, 23, 30, 33, 38, 53,55,59,64, 24, 32, 55, 59, 14 AY534389 5 10, 23, 25 30 3, 5, 10, 25, 59 23, 64 55, 55, 102 6 5, 10 5, 3, 11 13, 20 10 20 14 5 6 69, 72 4 25, 32, 71 30, 63 5 25, 54 100

Linnaeus, 1758 Gmelin, 1791 Linnaeus, 1758 Pease, 1865 Lorenz, 1985 Sowerby 1832 I, Melvill, 1888Melvill, Gmelin, 1791 Sowerby 1837 I, Sowerby1837 I, Melvill, 1888 Gray, 1825 Gray, 1825 Schilder & Schilder, 1929 102 Schilder & Schilder, 1938 Sowerby 1832 I, 5 Geret, 1903Geret, Perry, 1811Perry, Reeve, 1845 Melvill, 1888 Jousseaume, 1876 Linnaeus, 1758 Gray, 1825 1825 Gray, Linnaeus, 1758 Linnaeus, 1758 Melvill, 1905Melvill, Gaskoin, 1843 Gmelin, 1791 . cf cf. clandestina cf. (Andaman)

cf. bitaeniata cf. asellus cf. vespacea OTUs Authors Sampled OTUs Authors ) continued ( 42. Bistolida ursellus Bistolida hirundo Bistolida ursellus Bistolida ursellus stolida Bistolida stolida candida 37. Cypraeovula mikeharti Bistolida goodallii 40. Bistolida stolida diagues rubiginosa Bistolida stolida Bistolida erythraeensis Bistolida erythraeensis 41. Bistolida owenii Bistolida stolida clavicola clavicola Bistolida stolida Cypraeovula edentula Cypraeovula alfredensis 38. Cypraeovula algoensis Palmadusta contaminata distans Palmadusta contaminata contaminata Palmadusta asellus clandestina Palmadusta ziczacmisella Palmadusta diluculum Palmadusta clandestina passerina Palmadusta artuffeli (Andaman) Palmadusta humphreysii 39. Palmadusta clandestina Palmadusta asellus ziczac Palmadusta saulae Palmadusta lutea

154 MEYER ) continues ( AY161566 AY161800 AY161559 AY161793 AY161560 AY161794 AY161561 AY161795 AY161562 AY161796 AY161567 AY161801 71 71 23, 50, 98 23 AY534393 AY534470 AY534470 AY534393 23 98 3 38 25, 29 15, 24, 38, 52, 53,55,59, 5, 38, 53, 59, 73, 14, 5, 14, 19, 30, 38, 59, 64, 73, 76, 80, 14, Hidalgo, 1906 5, 8, 9 AY161542 AY161776 Kuroda, 1938 93 AY161516 AY161749 Melvill, 1888 Lorenz, 2002 Linnaeus, 1758 Iredale, 1930 Raybaudi, 1987 Lorenz, 2002 35 33 50 40 38 AY161571 AY534400 AY161805 AY534478 AY534401 AY161568 AY534479 AY161802 AY534402 AY534480 Schilder, 1933 Gmelin, 1791 29, 52 19, 28, 29 AY161540 AY161774 AY161538 AY161772 Melvill & Melvill Standen, 1904 Melvill, 1888 Gmelin, 1791 53, 59, 69, Melvill, 1888 Weinkauff, 1881 1888Melvill, Reeve, 1846 Schilder & Schilder, 1938 Burgess, 1993 Sowerby 1832 I, 54 70 Lorenz & Raines, 2001 Schilder, 1971 1811Perry, 73 75 Lorenz, 1999 73 Lorenz, 2002 67 69, 70 Gaskoin, 1849 1833Duclos, Smith, 1881 71 Lorenz, 2002 5 100 100 7 7 41 AY161575 AY534394 AY161809 AY534471 AY534395 AY161572 AY534472 AY161573 AY534396 AY161806 AY161807 AY534473 AY161574 AY534474 AY161808 AY534397 AY161565 AY534398 AY534475 AY161799 AY534476 AY161563 AY161797 AY161564 AY161569 AY161798 AY161803 Biraghi, &, Nicolay, 1993 64 AY161570 AY161804 Lorenz, 1989 3 Lorenz, AY534477 AY534399 australiensis australiensis

langfordi A B . abaliena . cf Accession # Accession OTUs Authors Sampled localities* 16S COI 16S localities* Sampled OTUs Authors chinensis chinensis exmouthensis (Andaman) Austrasiatica

) continued ( Bistolida kieneri kieneri Bistolida kieneri 51.Cribrarula exmouthensis magnifica Cribrarula cribraria cribraria melwardi 52.Cribrarula cribraria Cribrarula cribraria rottnestensis abrolhensis53.Cribrarula cribraria Nesiocypraea Bistolida kieneri depriesteri Bistolida kieneri Ovatipsa chinensis Bistolida kieneri depriesteri Bistolida kieneri 43. Ovatipsa chinensis amiges Ovatipsa coloba Talostolida teres Talostolida pellucens 44. Talostolida subteres 45. Talostolida latior Cribrarula gaskoini Cribrarula catholicorum 46. Cribrarula taitae Cribrarula cumingii 47. Cribrarula garciai Cribrarula astaryi Cribrarula cribraria comma 48. Cribrarula pellisserpentis 49.Cribrarula esontropia francescoi Cribrarula cribellum Cribrarula esontropia Cribrarula fallax Cribrarula cribraria Cribrarula gaspardi 50. Cribrarula cribraria Cribrarula cribraria

INCREASED MOLECULAR SAMPLING IN CYPRAEIDAE 155 ) continues ( # COI AY534490 AY534491 Accession 16S AY161608 AY161842 AY161610 AY161844 AY161612 AY161846 AY161611 AY161845 AY161606 AY161840 AY161603 AY161837 AY161577 AY161811 AY161515 AY161748 AY161514 AY161747 AY161609 AY161843 AY161601 AY161835 AY161584 AY161818 AY161604 AY161605 AY161838 AY161839 AY161600 AY161834 AY161578 AY161812 AY161607 AY161841 AY161602 AY161836 AY534412 AY534489 AY534405 AY534483 AY534408 AY534409 AY534486 AY534404 AY534404 AY534482 AY534410 AY534487 AY534411 AY534488 AY534407 AY534485 AY534406 AY534484 AY534415 AY534492 AY534403 AY534481 AY534416 AY534493 33, 53, 56 localities* localities* 11 31, 98 51 38 11 9 11 AY534414 5, 33, 14, 14, 32, 109 25 55, 16 107 25 1, 3, 5 1, 5 53, 54 54 16, 24, 30, 33, 59 16, 23 11,14, 16, 30, 52, 55,57, 58, 9, 10 9, 3, 14, 20, 57, 25 23, 30, 33, 38, 52,53,55,64 55, 67 51 51 55 104 95 Gray, 1824 Schilder, 1927 Schilder, 1964 Cate, 1969 Linnaeus, 1758 Roberts, 1913 Habe, 1970 Kuroda, 1960 Kuroda & Habe, 1961 1832 I, Sowerby 55 1810Lamarck, Reeve, 1835 Schilder & Schilder, 1938 1938 Schilder, & Schilder Schilder, 1937 Gray, 1824 Born, 1778 Schilder, 1927 Linnaeus, 1758 Iredale, 1939 1791Gmelin, Schilder & Schilder, 1938 Linnaeus, 1758 57 Gmelin, 1791 Born, 1778 Roding, 1798 Perry, 1811 3 3, Lorenz, 2002 AY534413 Schilder & Schilder, 1938 Melvill, 1901 69, 71 Lorenz & Sterba, 1999Lorenz & Sterba, 69, 70 hirasei sakurai

OTUs Authors Sampled OTUs Authors cylindrica Austrasiatica Austrasiatica

) continued ( Nesiocypraea 56. Erronea pallida 57. Erronea vredenburgi rabaulensis Erronea 58. 59. Erronea fernandoi onyx (Adusta) Erronea Palmulacypraea katsuae 54. Palmulacypraea musumea 55. Erronea xanthodon adusta (Adusta) Erronea Erronea (Adusta) subviridis Nesiocypraea Erronea (Adusta) subviridis dorsalis dorsalis subviridis (Adusta) Erronea Erronea (Adusta) onyx melanesiae 60. Erronea pyriformis Erronea cylindrica cylindrica Erronea Erronea ovum chrysostoma chrysostoma ovum Erronea Erronea caurica 61. Erronea cylindrica lenella ovum ovum Erronea 62. palauensis ovum Erronea errones Erronea Erronea caurica cf. derosa derosa cf. caurica Erronea Erronea caurica dracaena quinquefasciata caurica Erronea 63. Erronea caurica elongata 64. Erronea caurica spp. 1 65. Erronea caurica spp. 2 66. Erronea caurica samoensis Purpuradusta serrulifera minoridens Purpuradusta 67. Purpuradusta oryzaeformis

156 MEYER AY161576 AY161810 AY161579 AY161813 AY161580 AY161814 AY161583 AY161817 AY161598 AY161832 AY161599 AY161833 AY161585 AY161819 AY161595 AY161829 11, 51 18, 19, 23, 61, 70 98 56 98 12 12 92 14, 15, 20, 28, 33,59, 54, 19, 30, 14, 30, Gray, 1828 52, 59, Kiener, 1843 Iredale, 1939 5 23, 38, AY534417 AY534494 Gill, 1858 Gill, 1849 Gaskoin, 5 3, AY161581 AY161815 Gmelin, 1791 1845Mighels, Lorenz, 2002 Schilder, 1933 Sowerby 1832 I, 9, 14 5, 30, 31, 59, 64, 69, 71 73 14, 23, 38, AY161582 AY161816 AY534418 AY534419 AY534495 AY534496 Crosse, 1868Crosse, 14 AY161588 AY161822 Reeve, 1857 AY161820 Swainson, 1829 Sowerby 1888 III, Schepman, 1907 Linnaeus, 1771 Sowerby 1870 II, 14 22 Dautzenberg, 1906 22 106 10 64 1824 Gray, AY161588 23, 52, 53, 55, 59,64 69, 70 AY161586 5 AY161589 AY161823 AY534421 AY534420 AY534422 AY534498 AY534497 AY161590 AY534423 AY161824 AY534499 AY161587 AY161821 Hidalgo, 1906 1791Gmelin, Cate, 1963 Schilder & Schilder, 1938 1824Gray, Cox, 1873 33 Gaskoin, 1849 50 Souverbie,1865 5 3, Steadman & Cotton, 1943 1824 Gray, 67 24 16, 24, 27, 32, 52,57 15, 17 32, 52, 54 54 AY161592 AY161591 AY161826 AY534424 AY161825 AY534425 AY534500 AY534501 AY534426 AY161596 AY161593 AY534502 AY161594 AY161830 AY161827 AY161828 AY161597 AY161831 Schilder, 1958 92 AY534427 AY534503

A B microdon Accession # Accession (Andaman) OTUs Authors Sampled localities* 16S COI 16S localities* Sampled OTUs Authors ) continued ( Purpuradusta microdon microdon Purpuradusta Purpuradusta hammondae 68. Purpuradusta microdon chrysalis notata Purpuradusta gracilis Purpuradustafimbriata fimbriata Purpuradusta fimbriata unifasciata 69. Purpuradusta fimbriata marquesana 70. Purpuradusta fimbriata waikikiensis Contradusta walkeri Contradusta bregeriana bregeriana Contradusta 71. Contradusta? barclayi 72. Contradusta? pulchella 73. Notadusta? hungerfordi Notadusta martini Notadusta punctata punctata 74. Notadusta punctata trizonata Notadusta punctata Notadusta punctata berinii Notadusta punctata berinii Melicerona listeri 75. Melicerona melvilli listeri 76. Meliceronafelina Eclogavena dayritiana Eclogavena quadrimaculata thielei Eclogavena quadrimaculata quadrimaculata Eclogavena coxeni Blasicrura pallidula pallidula Blasicrura pallidula rhinoceros 77.Blasicrura vivia pallidula cf. Blasicrura interrupta 78. Blasicrura summersi