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NAUTILUS

Volume 133, Numbers 3-4

November 27, 2019

ISSN 0028-1544

CONTENTS

M. G. Harasewych The complete mitochondrial genome of Neptuneopsis gilchristi G.B.

Makiri Sei Sowerby III, 1898 (Neogastropoda: Volutidae: Calliotectinae) .......0.0.0.0.c. 67

Herman H. Wirshing

Vanessa L. Gonzalez

Juan E. Uribe

Robert A. Krebs Post-glacial dispersal patterns of Pyganodon grandis (Bivalvia: Unionidae)

Lyuba E. Burlakova HaNWS) Have lkowmere Gireatt Lalas WRAteHSl OG! <cocosadocondnséonsdecoodenccnssoGbbodesosoosnsdeexe000060000 74

David T. Zanatta

Roland Houart Description of two new muricid species (Gastropoda: Muricidae:

Muricopsinae) from the western Atlantic and the eastern Pacific ......0.0.0.00... 85

Jinxiang Jiang Description of two new species (Bivalvia: Vesicomyidae, Verticordiidae)

Yaqin Huang frOmparcoldasceppnEthers Outhg Chinayscalessmaceet ste ass en een eee Rete 94

Qianyong Liang

Junlong Zhang

ANTELVETP TIGERS scosSassansnasooace soodoneods saSebe note SeIGiia a cEaa a SS BU CGS oc REASON Ie AT CSAS Ae 103

\TH SONA

DEC 03 2019 }

15.

16.

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THE NAUTILUS 133(3-4):67-73, 2019

Page 67

The complete mitochondrial genome of Neptuneopsis gilchristi

G.B. Sowerby III, 1898 (Neogastropoda: Volutidae: Calliotectinae)

M. G. Harasewych'

Makiri Sei

Herman H. Wirshing

Department of Invertebrate Zoology, MRC-163

National Museum of Natural History

Smithsonian Institution

P.O. Box 37012

Washington, DC 20013-7012 USA

ABSTRACT

We report the complete mitochondrial genome of Neptu-

neopsis gilchristi G.B. Sowerby III, 1898, the type species of

the monotypic genus Neptuneopsis. This mitogenome is

15,312 bp in length and has a GC content of 31.3%. The gene

order of the 13 protein-coding genes, 2 ribosomal RNA genes,

and 22 transfer RNA genes, is identical to that of most neo-

gastropods other than of several conoidean taxa, which differ

only in the location of one or more tRNA genes. The potential

origin of replication is located in a 127 bp non-coding region

between tRNA-Phe and COX3 that has a high A + T content

(77.9%). Phylogenetic analyses using maximum likelihood and

Bayesian inference with nucleotide sequences of all protein-

coding and ribosomal genes show Neptuneopsis to be sister to

the few volutid species for which complete or partial mito-

genome data are available. Neptuneopsis gilchristi has an

operculum and a triserial radula, while the ayia volutids in our

analyses have lost the operculum and have a derived uniserial

radula.

Keywords: Gastropoda, mitogenome, phylogeny, gene order

INTRODUCTION

The neogastropod family Volutidae comprises a group of

predatory marine gastropods that inhabit sand and mud

substrates from the intertidal zone to abyssal plains, from

the tropics to polar seas. Like many neogastropod families,

Volutidae has origins in the early Late Cretaceous

(Cenomanian) (Stephenson, 1952: Pojarkov a, 1984:

Tracey et al., 1993; Fossilworks, 2019). The World

living and fossil species-level taxa and 115 living and fossil

genus-level taxa attributed to this family. Pilsbry and

Olsson (1954) reviewed the early taxonomic history of

Volutidae and partitioned the family into 12 subfamilies

1 - :

Corresponding author: [email protected]

Vanessa L. Gonzalez

Global Genome Initiative

National Museum of Natural History

Smithsonian Institution

P.O. Box 37012

Washington, DC 20013-7012 USA

Juan E. Uribe

Department of Invertebrate Zoology,

MRC-163

National Museum of Natural History

Smithsonian Institution

P.O. Box 37012

Washington, DC 20013-7012, USA

and 8 tribes, acknowledging this to be a tentative effort.

The most recent classification (Bouchet et al., 2017:349)

recognizes 10 subfamilies (2 extinct) and 11 tribes.

meee classifications based on morphological and an-

atomical data (e.g., Thiele, 1929; Wenz, 1943) included

Volutidae in the superfamily Volutoidea, together with the

families Olividae, Mitridae, Tima bral ErD. Harpidae,

Marginellidae, and Cancellariidae, while subsequent

dias Reiter have variously distributed these families

among the rachiglossan neogastropods (see Harasewych

et al., 1997: fig. : 2). The most recent classification (Bouchet

et al., 2017: 349, 379) based on molecular data (Fedosov

et al., 2015) includes only Volutidae and Cancellariidae in

Volutoidea, with the remaining families either elevated to

superfamilies or unassigned to superfamily.

The family Volutidae has been sparsely represented

in morphological and molecular investigations of neo-

gastropod phylogeny. In this study, we report the com-

plete mitochondrial genome of Neptuneopsis gilchristi

G.B. Sowerby HI, 1898. a member of the subfamily

Calliotectinae. It represents the second complete mito-

genome of a volutid to be determined. Features such as

genome length and gene order are compared to those of

online neogastropods, and the phylogenetic position of

Neptuneopsis within Volutidae is inferred under both

maximum likelihood (ML) and Bayesian frameworks

based on mitochondrial protein-coding and RNA genes.

MATERIALS AND METHODS

DNA Extraction and Sanger Sequencing: Genomic

DNA (gDNA) was extracted from a 40 mg section of

proboscis wall obtained from an alcohol-preserved

specimen of Neptuneopsis gilchristi [Natal Museum,

V1106; South Africa, South of Cape St. Blaize (34°47 S,

22°10 E), dredged in 97 m, Stn. A17383, NMDP, R/V

AFRICANA, 6 May 1995] using the AutoGenprep 965

(Autogen, Holliston, MA, USA). Manufacturer -provided

proteinase K (Autogen) was used for initial tissue lysis,

Page 68

which was run overnight at 56°C with continuous agita-

tion. Portions of cytochrome c oxidase I (COX) and 16S

rRNA genes were PCR-amplified and Sanger-sequenced

using the primers and protocols in Harasewych (2018).

These two mitochondrial gene fragments were used as

scaffolds for the assembly of the N. gilchristi mitogenome

(see Assembly below).

Library Construction and Illumina Sequencing:

Extracted gDNA was visualized on a 1.5% agarose gel, and

quantified using a Qubit dsDNA HS Assay Kit (Ther-

moFisher, Pittsburg, PA). After quantification, g DNA was

sonicated using the Covaris ME220 with microtube-50

AFA fiber screw-caps (Covaris, Woburn, MA) targeted for

350bp fragments. Sonicated gDNA was then cleaned

using Kapa Pure Beads (KAPA Biosystems, Wilmington,

MA) at a 0.9X beads-to-sample ratio, which targeted

fragments >250bp. Size-selected gDNA was then quan-

tified with Qubit dsDNA HS Assay Kit, and the Agilent

2200 TapeStation (Agilent, Santa Clar a, CA) was used to

validate gDNA size sclecion.

Total gDNA libraries were prepared using the NEB-

Next Ultra II DNA Library Prep Kit for [lumina together

Table 1.

THE NAUTILUS, Vol. 133, Nos. 3-4

with the NEBNext Multiplex Oligos for Illumina (New

England BioLabs, Ipswich, MA). Size selection of

adaptor-ligated libraries (400-500 bp) and adaptor/PCR

cleanups were performed using Kapa Pure Beads, and

library size was validated with the Agilent 2200 Tape

Station. Libraries were quantified using qPCR (ViiA 7,

ThermoFisher) to ensure generation of adaptor-ligated

libraries. A 4nM library concentration was denatured for

clonal amplification and sequenced on an Illumina MiSeq

(IIhimina, San Diego, CA) with MiSeq Reagent Kit v3 at

the Smithsonian National Museum of Natural History's

Laboratories of Analytical Biology.

Assembly: Low quality reads were removed using

TrimGalore v. 0.6.3 dev (https://github.com/FelixKrueger/

TrimGalore), with thresholds for minimum Phred scores

set to 20 and minimum read lengths set to 20 bp. The N.

gilchristi mitogenome was assembled using Geneious

pemee 2019.2 3 .1 (http:/www.geneious.com) using the

following protocol: Sanger- -sequenced COXI and 16S

rRNA gene fragments were used as scaffolds using the

“map to reference” tool with “minimum overlap iden-

tity’ set to 98-99% and “minimum overlap” set to

List of taxa used in phylogenetic analyses, their GenBank accession numbers, source of sequence data, and size of the entire

mitogenome. Taxon names as well as their superfamily and family assignments have been updated according to WoRMS (2019).

* Indicates that only a partial mitogenome sequence was available.

GenBank No. Source

length (bp)

Superfamily Family Taxon

OUTGROUPS - LITTORINIMORPHA

Stromboidea Strombidae Lobatus gigas

Naticoidea Naticidae Naticarius hebraeus

Tonnoidea Cymatiidae Monoplex parthenopeus

Tonnoidea Charoniidae Charonia lampas

Tonnoidea Cassidae Galeodea echinophora

NEOGASTROPODA

Volutoidea Cancellariidae Bivetiella cancellata

Volutoidea Volutidae Neptuneopsis gilchristi

Volutoidea Volutidae Cymbium olla

Volutoidea Volutidae Alcithoe lutea

Volutoidea Volutidae Alcithoe benthicola

Volutoidea Volutidae Amoria hunteri

Volutoidea Volutidae Cymbiola pulchra

Muricoidea Muricidae Bolinus brandaris

Muricoidea Muricidae Rapana venosa

Muricoidea Muricidae Reishia clavigera

Olivoidea Ancellariidae Amalda northlandica

Unassigned Babyloniidae Babylonia areolata

Unassigned Babyloniidae Babylonia lutosa

Buccinoidea Columbellidae Columbella adansoni

Buccinoidea Nassariidae Tritia reticulatus

Buccinoidea Buccinidae Neptunea arthritica

Buccinoidea Buccinidae Buccinum undatum

Conoidea Conidae Californiconus californicus

Conoidea Conidae Conus borgesi

Conoidea Conidae Conus textile

Conoidea Terebridae Oxymeris dimidiata

Conoidea Fussiturridae Fusiturris similis

Conoidea Turridae lotyrris cerithiformis

Conoidea Turridae Gemmuloborsonia moosai

NC_024932 Marques et al 2014 15,461

NC_028002 Osca et al. 2015 15,384

NC_013247 Cunha et al. 2009 15,270

NC_037188 Cho et al. 2017 15,330

NC_028003 Osca et al. 2015 15,388

NC_013241 Cunha et al. 2009 16,648

MN125492 This study 15,312

NC_013245 Cunha et al. 2009 15,375

JN182219 Hills et al. 2011 7,689 *

JN182217 Hills et al. 2011 7,692 *

JN182226 Hills et al. 2011 Ces ©

JN182216 Hills et al. 2011 1083

NC_013250 Cunha et al. 2009 15,380

NC_011193 Sun & Yang 2014 IS ATA

NC_010090 Ki et al. 2010 15,285

NC_014403 McComish et al. 2010 15,354

NC_023080 Chen & Ke, unpublished 15,445

NC_028628 Xiong et al. 2015 15,346

KP716637 Osca et al. 2015 16,272

NC_013248 Cunha et al. 2009 15,271

KU246047 Hao et al. 2016 15,256

NC_040940 Jonsson et .al. 2019 15,265

NC_032377 Uribe et al. 2016 15,444

NC_013243 Cunha et al. 2009 15,536

NC_008797 Bandyopadhyay et al. 2008 15,562

NC_013239 Cunha et al. 2009 16,513

NC_013242 Cunha et al. 2009 15,595

NC_008098 Bandyopadhyay et al. 2006 15,380

NC_038183 Uribe et al. 2018 15,541

$$$

M.G. Harasewych et al., 2019

Page 69

30-40 bp. An initial iterate, using the “fine tuning”

option tab, was set to 5X to confirm adequate initial

assembly to scaffolds. The 16S rRNA fragment did not

scaffold well, and, therefore, only COXI was used for

subsequent assembly iterations. Three-five iterations of

3X and 40X were needed before an assembled contig

of >15kb, the mitogenome size-approximation based on

related taxa, was acquired. The size and sequence of the

non-coding region, between the tRNA-Phe and COX3,

was confirmed using standard PCR and Sanger se-

quencing, with primers derived from within the flanking

genes: NEPTtrn-Phe F — 5’ GGT GGT AAA CAT ATC

TTA AGA TAG G = 3) and NEPIcox3 R—5' AAG TAA

GTG GAA TGG ATT ACG TCT C.

Genome Annotation: Mitochondrial elements were an-

notated using MITOS (Bernt et al., 2013), ARWEN (Laslett

and Canbiick, 2008) and the ORF finder in Geneious.

Phylogenetic Analyses: All 13 protein-coding and both

ribosomal genes from selected neogastropod and out-

group mitochondrial genomes (Table 1) were used to

determine the phylogenetic relationships of N. gilchristi.

Gene alignments were run in MAFFT v7 (Katoh et al.,

2019) and ambiguous positions removed using GBlocks,

v.0.91b (Castresana, 2000). The best-fit evolutionary

models and partition scheme were selected using Mod-

elFinder (Kalyaanamoorthy et al., 2017) through IQ-

TREE v.1.6.1 (Nguyen et al., 2014) with the option

Table 2. Position (start, stop), strand direction (+/-) and lengths (bp) of the genes in the mitochondrial genome of Neptuneopsis

gilchristi G.B. Sowerby HI, 1898 [15,312 bp; GenBank MN125492], initiation and termination codons (Init/Term) for protein-coding

genes, as well as their amino acid sequence lengths (Laa). Standard abbreviations for protein coding genes are used. Both three and one

letter abbreviations are listed for tRNA’s, along with the codon used. The numbers of intergenic nucleotides (Ign) are shown. Negative

numbers indicate overlap of genes.

Gene Start Stop Strand Directionn Length (bp) Init /Term Laa Ign

COX1 1 1,542 + 1,542 ATG/TAA 513 1]

COX2 1,554 2, 240 + 687 ATG/TAA 228 -3

tRNA-Asp (D) (gte)+ 2,238 2,308 4 71 0

ATPS 2,309 2, 467 + 159 ATG/TAA 52 5

ATP6 2.473 3,168 + 696 ATG/TAG 231 31

tRNA-Met (M) (cat)* 3,266 3,200 = 67 =3

tRNA-Tyr (Y) (gta) + 3,330 3,264 _ 67 31

tRNA-Cys (C) (gca)* 3,395 3,332 — 64 -2

tRNA-Trp (W) (tea) + 3,461 3,394 — 68 -2

tRNA-Gln (Q) (ttg)+ 3,525 3,460 - 66 0

tRNA-Gly (G) (tee)+ 3,594 3,526 = 69 -2

tRNA-Glu (E) (tte)+ 3,659 3,593 - 67 0)

12S rDNA 3,660 4.615 = 956 0

tRNA-Val (V) (tac)+ 4.616 4.683 ar 68 0)

16S rDNA 4.684 6,035 + 1,352 0)

tRNA-Leu (L1)(tag)+ 6,036 6,106 + 71 0)

tRNA-Leu (L2)(taa)* 6,107 6,175 + 69 0)

NAD1 6,176 lel) + 942 ATG/TAA 313 0

tRNA-Pro (P) (tgg)* 7,118 7,189 + 72 0

NAD6 7,190 7,690 + 501 ATG/TAA 166 5

CYT B 7,696 8,835 = 1140 ATG/TAA 379 6

tRNA-Ser(S2)(tga)+ 8,842 8,909 + 68 10

tRNA-Thr (T) (tgt)* 8,984 8,920 — 65 9

NAD4L 8,994 9,290, + 297 ATG/TAG 98 17

NAD4 9,308 10,657 + 1350 ATT/TAG 449 2

tRNA-His (H)(gtg)+ 10,660 10,726 + 67 26

NAD5 10,753 12.447 + 1,695 ATT/TAA 564 10

tRNA-Phe (F)(gaa)+ 12,458 12.5244 + 67 127

COX3 12,652 13,431 + 780 ATG/TAA 259 28

tRNA-Lys (K) (ttt)+ 13,460 13,528 =f 69 4

tRNA-Ala (A) (tgc)* 13,533 13,600 + 68 11

tRNA-Arg (R)(teg)* 13,612 13,680 + 69 6

tRNA-Asn (N)(gtt)* 13,687 13,754 + 68 §

tRNA-Ile (I) (gat)+ 13,763 13,830 =f 68 3

NAD3 13,834 14,187 ar 354 ATG/TAG 118 0

tRNA-Ser (S1)(gct)* 14,188 14,255 + 68 0)

NAD2 14,256 29 + 1086 ATG/TAA 361 -29

For tRNA’s * indicates that ARWEN the same as MITOS.

For tRNA’s + indicates that ARWEN selected over MITOS.

Page 70

“m TESTONLYMERGE” and Bayesian Information

Criterion (BIC).

Phylogenetic analyses, using a data matrix concatenated

in Geneious that included 12,636 nucleotide positions,

were performed with MrBayes v3.1.2 (Ronquist and

Huelsenbeck, 2003) running four MCMC chains for two

million generations, sampling every 1,000 and discarding

the first 25% as burn-in. Convergence of the trees was

determined using TRACER v1.6 (Rumbaut et al., 2007);

and IQ-TREE v1.6.1 using a combination of rapid hill-

climbing and stochastic perturbation methods with a total

of 1,000 pseudoreplicates of bootstrap to assess robust-

ness of the inferred tree.

RESULTS

Genome Content and Organization: The Illumina

sequencing run produced a total of 51,544,678 reads for

this species. After filtering and removing low-quality data,

50,964,146 reads (Phred scores = 20, length = 20 bp)

remained with an average length of 193.2 bp (SD 77.8). Of

these 12,954 mapped to the mitochondrial genome.

Coverage ranged from 115X to 345X per site (mean =

213.5: SD = 38.1).

15,312

tRNA - Ser1

tRNA - Ile

tRNA - Arg

tRNA - Lys

Neptuneopsis gilchristi

15,312 bp

11,500

NAD 4L

tRNA - Ser2

oof

tRNA - Thr

tRNA - Pro

THE NAUTILUS, Vol. 133, Nos. 3-4

The mitochondrial genome of Netuneopsis gilchristi

reconstructed from these data is a double-stranded circular

molecule 15,312 bp in length (GenBank Acc. No.

MN125492), composed of 31.0% A, 37.7% T, 15.8% C, and

15.6% G. It contains 13 protein-coding genes, two ribo-

somal RNA genes and 22 tRNA genes (Table 2, Fig. 1). Of

these, 29 genes are coded on the heavy strand (+ strand)

and only § tRNA genes, including the cluster MYCWQGE

(tRNA-M, tRNA-Y, tRNA-C, tRNA-W, tRNA-Q, tRNA-G,

and tRNA-E) and tRNA-T are coded on the light strand

(— strand). The gene order in N. gilchristi corresponds to

the consensus gene order shared by most caenogastropod

(Osca et al., 2015: 122) and neogastropod (Cunha et al.,

2009: 210, Fig. 1) mitogenomes. Known exceptions to this

gene order are limited to unrelated tRNA translocations

and inversions within Conoidea (Cunha et al., 2009; Uribe

et al., 2016: fig. 1; Uribe et al., 2018: fig. 2).

Gene overlaps total 41 bp at 6 gene junctures, the

longest (29 bp) between NAD2 and COX1. There are 19

intergenic regions (350 bp in total, 2.3 % of the mito-

genome) ranging from 2 to 127 bp, the largest between

tRNA-Phe and COX3 (Table 2).

The potential origin of replication (POR) is located in a

127 bp non-coding region between tRNA-Phe and COX3,

which has a high A + T content (77.9%).

500

tRNA - Met—@@=— tRNA - Tyr

tRNA - Gina

tRNA - Gly J

tRNA - Glu

tRNA - Leu1

tRNA - Leu2

Figure 1. Map of the mitochondrial genome of Neptuneopsis gilchristi (GenBank Acc. no. MN125492). Arrows indicate the direction

of transcription. Protein-coding genes are in green, ribosomal genes in red and transfer RNA genes in purple.

M.G. Harasewych et al., 2019 Page 71

Lobatus gigas

Naticarius hebraeus

Bivetiella cancellata 1 Cancellariidae | I

Monoplex parthenopeus

4001 Charonia lampas

100/1 Galeodea echinophora

Neptuneopsis gilchristi

tn Cymbium olla

1001 -— Alcithoe lutea

ait Alcithoe benthicola

93/1 Amoria hunteri

10011 Cymbiola pulchra

10011 Bolinus brandaris

Rapana venosa

10/1 Reishia clavigera

Amalda northlandica

eeUee Babylonia areolata

io0iL— Babylonia lutosa

i Columbella adansoni

59/0.99 Tritia reticulatus

99/1 Neptunea arthritica

400/1 Buccinum undatum

520.98] 499 Californiconus californicus

Conus borgesi

10011 Cylinder textile

Oxymeris dimidiata

Fusiturris similis

| TONNOIDEA

400/1

Volutidae

VAGIOLMO/N

‘

| MURICOIDEA

I Ancillariidae

| Babyloniidae

100/1

BUCCINOIDEA

YdOdOuLSVOOAN

0.9 100/ CONOIDEA

100/1 Lophiotoma cerithiformis

5410.98 Gemmuloborsonia moosai

Figure 2. Phylogenetic relationships of Neptuneopsis gilchristi based on maximum likelihood and Bayesian analyses of nucleotide

sequences of mitochondrial protein-coding and ribosomal genes. Branch support shown as maximum likelihood bootstrap values

(when = 50) / Bayesian posterior probability (when = 0.7).

Protein coding genes comprise 73.3% of the entire

genome (11,229 bp). The most common start codon is

ATG, occurring in all genes except ND4 and ND5, which

use the ATT start codon. The stop codon TAA is used in

nine genes and the TAG stop codon in four genes. Both

the large ribosomal gene (16S rRNA, 1352 bp) and the

small ribosomal gene (12S rRNA, 956 bp) (total =

2,308 bp, 15.1% of mitogenome) occur on the heavy

strand. Of the 22 t-RNAs, 14 are on the heavy strand, 8 on

the light strand. The total length of the tRNAs is 1496 bp,

9.8% of mitogenome.

Phylogenetic Analyses: Phylogenetic analyses of the

concatenated nucleotide sequences for all protein-coding

and ribosomal RNA genes using both maximum likelihood

and Bayesian inference each produced a single tree that

was fully resolved, and in which most nodes were well

supported. The two trees were congruent except for the

relative positions of Oxymeris and Fusiturris, among the

non-conid Conoidea (Figure 2).

DISCUSSION

The mitogenome of Neptuneopsis gilchristi (15,312 bp) is

63 bp shorter than that of Cymbiwm olla, the only other

volutid for which a complete mitogenome is known. The

order and strand orientation of the mitochondrial genes is

the same for both species, and also matches the portion

(>7,600 bp) of the mitogenome between ND3 and tRNA-

Leu2 that has been reported for 11 species of Alcithoe,

Amoria hunteri, and Cymbiola pulchra (Hills et al., 2011:

Table 1, Fig. 1). The ND2 gene overlaps the COX] gene

by 29 bp for all volutid species reported to date except

Cymbium olla, which has a 17 bp gap between the two

genes.

Analyses using both maximum likelihood and Bayesian

inference (Figure 2) strongly support the monophyly of

the superfamilies Conoidea, Buccinoidea, Muricoidea,

and Tonoidea. The family Volutidae is also recovered as

monophyletic with strong support, but not the super-

family Volutoidea, as the sole cancellariid in the dataset

appears more closely related to Tonnoidea, although

without significant support. A similar result was obtained

by Cunha et al. (2009: fig. 3) and Osca et al. (2015: fig. 2)

when analyzing nucleotide sequences of complete mito-

chondrial genomes. The monophyly of Neogastropoda has

often been contradicted in multiple molecular studies (for

review, see Ponder et al., 2008: 368), as Tonnoidea typ-

ically are included among Neogastropoda (Osca et al.,

2015; Strong et al., 2019).

Neptuneopsis emerges as the earliest branching of the

few volutid taxa for which data on a significant portion of

the mitogenome are available. This is concordant with

relationships based on morphology, as Neptuneopsis is the

Page 72

THE NAUTILUS, Vol. 133, Nos. 3-4

only taxon within this study that retains an operculum and

a triserial radular ribbon, both considered plesiomorphic

features within Volutidae (Pilsbry and Olsson, 1954). Of

the remaining volutids, Amoria hunteri and Cymbiola

pulchra, members of separate tribes within the subfamily

Amoriinae, emerge as sister taxa, as do the congeners

Alcithoe lutea and A. benthicola.

However, the two species of Alcithoe do not form a

clade with Cymbium olla, although all three are classified

in the subfamily Cymbiinae. It is unclear if this discrep-

ancy may be due to the comparison of complete versus

partial mitogenomes. Cunha and colleagues (2009: 210)

noted the uneven contributions of various genes to

phylogenetic resolution, with COX2, ATP6, and NAD4

being rated the highest, and COXI and ATPS the lowest.

We regard this mitogenome as being an incremental

contribution toward a better understanding of the evo-

lutionary history of Volutidae.

ACKNOWLEDGMENTS

We thank Dai Herbert, Linda Davis, and Igor Muratov of

the KwaZulu-Natal Museum for making available the

specimen used in this study, as well as for providing

additional information and images. All laboratory work,

sequencing and analyses were conducted in and with the

support of the Laboratories of Analytical Biology (LAB)

facilities of the National Museum of Natural History.

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THE NAUTILUS 133(3-4):74-84, 2019

Page 74

Post-glacial dispersal patterns of Pyganodon grandis (Bivalvia:

Unionidae) into the lower Great Lakes watershed

Robert A. Krebs

Department of Biological, Geological,

and Environmental Seienees

Cleveland State University

Cleveland, OH 44115-2406 USA

[email protected]

Great Lakes Center

SUNY Buffalo State

Lyuba E. Burlakova

Buffalo, NY 14222 USA

David T. Zanatta

Department of Biology

Institute for Great Lakes Research,

Central Michigan University

Mount Pleasant, MI 48859 USA

ABSTRACT

Pyganodon grandis (Say, 1829) and other unionid mussels arrived

in the Laurentian Great Lakes only after the last glacial maximum

in North America. The mussel assemblage is thought to have

entered Lake Erie as a wave of expansion ‘Gon west to east, and

moved upstream within tributary rivers. A similar process for most

mussels occurred in Lake Ontario, but by eastern species, as the

two lake systems are separated by Niagara Falls. Only P. grandis is

abundant in both lakes. We applied variation in a fragment of the

mtDNA Carboxylase I (COI) gene to identify potential historical

paths for P. grandis to enter these watersheds. Nearly complete

monomorphy characterized the Lake Ontario population for the

most common Lake Erie allele, which is concordant with se-

quential founder effects across the Great Lakes. A gy including

populations from both Lakes, the intervening Niagara River,

tributaries of Lake Erie, and additional samples from below the

Laurentian Divide, was 0.10. The southern tributaries of Lake

Erie had greater genetic diversity, although few haplotypes were

shared among regions or even neighboring streams. Tributary and

lake populations differed significantly, yet variation across this

divide was not significant. This pattern likely arose from stream

capture of upper Ohio River tributaries that once flowed south,

and then population isolation, with upstream dispersal from Lake

Erie limited to the lower reaches.

INTRODUCTION

Population structure and _ historical biogeography of

freshwater mussels (Unionidae) is often difficult to assess

because current populations may be reduced and disjunct

(Berg et al., 2007; Zanatta and Harris, 2013), causing

many species to be endangered at a state or federal level

(Haag and Williams, 2014; Johnson et al., 2018). Common

species may therefore be more useful to trace distribution

patterns (Mynsberge et al., 2009), especially where ge-

netic markers are ‘applied (Gillean et al. 2007, 2008:

Hewitt et al., 2018; Mathias et al., 2018). Previously,

mtDNA sequence data were generated for one such

common. species, Pyganodon grandis (Say, 1829), in

Canada (Cyr et al., 2007; Does Beaupré et al., 2012),

and across the upper Mississippi watershed and into Lake

Erie. Low levels of spatial variation were identified across

these once glaciated regions, except for one possible

isolated northern glacial refugium in Minnesota that

remained unconnected to subsequent colonization of

the Great Lakes (Krebs et al. 2015). As P. grandis is

thought to be a host generalist, this mussel likely followed

diverse fishes that spread rapidly into new watersheds

(Bernatchez and Wilson, 1998), and it persisted through

ecosystem changes that later eliminated or nearly clttaatt

nated a great many other species (Zanatta et al., 2015).

While P. grandis tends to be a small component of mussel

assemblages i in flowing waters, its ability to colonize small

ponds and headwaters (Smith et al., 3002) presents op-

portunities to move between watersheds

Of the Unionidae, only P. grandis is abundant in both

lakes Erie and Ontario. Lake Ontario! is separated from Lake

Erie to the northeast by the 60 km long Niagara River. The

lake level falls from 174 m to atvont 76 m in this short

distance with a 70 m drop at Niagara Falls and a series of 5

gorges proceeding down river, forming an effective eco-

logical barrier. The lower elevation coupled with a slight

increase in latitude make the regions ecologically similar

except that Lake Erie is shallower to the west and Lake

Ontario to the east (Bossenbroek et al., 2018: Hoffman et al.

2018). The Appalachian Mountains extend below the lakes

forming a sharp front facing east, and the Allegheny Plateau

to the west, which form an easter biogeographic divide.

More subtly, a low ridgeline developed just south of Lake

Erie, separating the south-flowing Ohio River watershed

from the Great Lakes, which all empty north through Lake

Ontario and the St. Lawrence Seaway (Lewis et al., 2012).

Numerous Lake Erie tributaries possess a diverse yet varied

mussel assemblage above a low fall line that forms as the

rivers descend the Allegheny Plateau (Lyons et al., 2007),

causing upstream (Kebs et al., 2010a, 2013) and near-lake

(Crail a al., 2011; Zanatta et al., 2015: Krebs et al., 2018)

assemblages to differ. Nonetheless, mixing of various aquatic

groups can occur (Rahel, 2007).

We followed up those earlier studies to investigate two

knowledge gaps in the genetic structure in P. grandis within

the Gist Lakes: what was the likely source of populations in

Lake Ontario on the eastern edge of the St. Lawrence-Great

R.A. Krebs et al., 2019

Page 75

Lakes biogeographical province (Haag, 2012), and are Lake

Erie tributary population related more to the lake pop-

ulations or to populations in adjacent lotic watersheds. This

latter question considers possible historical connectivity be-

tween tributaries separated by the Laurentian Divide. Sep-

arately, given that an enigmatic and highly divergent (by 9%

of amaleottale bases) haplotype form exists within some P.

grandis populations (Cyr et al., 2007; Doucet-Beaupré et al.,

3012), we look at how new data can explain whether these

variant haplotypes are of hy brid origin or are historic within

the Pyganodon lineage and finally soma on how patterns of

male-inherited mitochondrial variation contrast with pat-

terns in the classically female-inherited form (Kr ebs, 2004).

MATERIALS AND METHODS

Over many years, small numbers of individuals, as re-

stricted by permits, were collected across various Ohio

Pelican L.

fe)

RiceR.°

Maumee

Blanchard (8) a

a Sandusky

Figure 1.

Lake Sainte

Malis

Lake Ontario

ve cot River \)

i ‘Lake ke Erie

Eagle Ck

cae Black R =

-. Killbuck

streams (Figure 1) initially as whole specimens where

Pyganodon grandis was common and later just mantle

clips during TER surveys within inlets and river mouths

along the southem Great Lakes (Krebs et al., 2010a;

Zanatta et al. , 2015: Bossenbroek et al., 2018). Here we

collated previously published CO1 sequence data from

Lake Erie (N=245), regions west of Lake Erie (N=94),

and the Niagara River (N= 55) (Krebs et al., 2015), and

generated new P. grandis CO] sequences (T Table 1) from

the Lake Onno region (N=66), Lake Erie tributary

steegns (N=8i7)k aiid Ohio River tributary streams

(N=20), along with 28 CO] sequences of the male-

alienate aiiticrlnomnchsel lineage.

Total DNA extraction, PCR methods and analyses were

described previously for standard female- inherited CO1

barcode sequences and for a small sample of male-

inherited sequences that were useful to confirm species

identification where female sequences were highly variant

Vermilion Cuyahoga

80 100km

Sampling region of P. gr andis for the present study, denoting the Great Lakes and five surrounding sites where rare

anomalous Type A ae conde V. mane were recovered. The inset shows the rivers sampled with number of individuals se quenced for

the various Ohio drainages of Lake Erie, and two additional streams, Eagle and Kilbuck Creeks, which comprise part of the Ohio River

headwaters. The Laurentian Divide separating these biogeographic regions is indicated by the dashed line.

Page 76

Table 1.

THE NAUTILUS, Vol. 133, Nos. 3-4

Genetic diversity in P. grandis across the five defined geographic areas, samples from within Lake Erie, the Niagara River,

Lake Ontario, the Tributaries of Lake Erie (LE), and for samples from two headwater tributaries of the Ohio River, Eagle Creek and

Kilbuck Creek in Ohio. Reported are sample size (N), allele number, polymorphic sites, haplotype diversity (H) or the likelihood that two

sequences drawn at random will differ, 7, the mean sequence variation among all haplotypes within a population, @,, the estimation of

4N based on the number of segregating sites, 0, the estimation of 4Nu based on nucleotide differences, and Tajima’s D, a test of the

difference between 0, and @,. which is predicted to be equivalent under neutral evolution.

Region N Allele Number Polymorphic sites

Lake Erie 245 29 31

Niagara R. 5D 9 §

Lake Ontario 66 4 5

LE tributaries 87 28 72

LE tributaries’ 78 24 2D

Ohio R. tributaries 20 6 61

5 7

‘results omitting the highly variant Type A haplotypes

** P< (0.01, *** P < 0.001

Ohio R. tributaries? 19

(Krebs, 2004; Krebs et al., 2015). All haplotype identifi-

cation codes correspond to the latter paper and Genbank

(Table 1). Sequences of individuals were entered into

DnaSP V 5.1 from which haplotype networks were

constructed in Network V 4.6.1 (Rohl, 2004). Poly-

morphic sites, transitions, and transversions were

weighted equally, although transitions were 6-fold more

common and almost every variant was a silent site. A

phylogeny was constructed in MEGA7 (v.7.0.26) under

maximum likelihood using the Kimura algorithm. The

ae topology was exported and drawn in FIGTREE

(http://tree.bio.ed.ac.uk/software/figtree/). To contrast bio-

geographic regions in an Analysis of Molecular Variance

(AMOVA), all Lake Erie sequences were pooled, and they

were assessed against all samples from Lake Ontario, and

all Lake Erie neboterny rivers. The samples from below

the Laurentian Divide were pooled as a separate group.

Among these sets, pairwise @gy analyses and a test of

neutrality, i.e., Tajima’s D Ae 1989), were run in

ARLEQUIN version 3.5.1.2, using 30,000 permutations

for tests of significance silane with mismatch analysis as a

separate test of population expansion (Excoffier and

Lischer, 2010). Tajima’s D contrasts @,, the estimation of

4N based on the number of segregating sites, and 0,

the estimation of 4Npz based on nucleotide differences,

which are predicted to be the same under neutral evo-

lution and a stable population size.

RESULTS

Expanding the CO1 data set for P. grandis to include Lake

Ontario, Lake Erie tributaries and two streams south of the

Laurentian Divide (Table 1) combined for 31 additional

haplotypes based on just a short mtDNA sequence

(Figure 2, accession numbers MN125095- MN125125).

The Lake Ontario samples were almost monomorphic for

the common haplotype, at 94% H1, with only two unique

haplotypes collected and one other haplotype that was

shared with the Niagara River samples. This common lake

haplotype also was the most frequent one in streams, but at

a frequency below 50%, making stream samples much

Diversity H a7 x 100 0, 0, Tajima’s D

0.31+0.04 0.085 5.10 0.534 —250F5*

0.27+0.08 0.057 1.74 0.362 —2.13**

0.12+0.05 0.029 1.05 0.181 —1.90**

0.77+0.05 1.940 14.29 12.22 —0.48

0.72+0.06 0.186 4.46 1.17 —2, 23t**

0.73+£0.07 1.20 17.19 7.579 —2.27***

0.70+0.08 0.316 2.00 1.988 —0.02

more genetically variable than lake samples. For other

multicopy haplotypes, most (6 of 8) occurred in only one

stream, or only in lake samples (12 of 13 multicopy hap-

lotypes), while only 1 was found both in Lake Erie samples

and in a tributary. Similarly , just one haplotype was shared

between tributaries separ Aiea by the Laurentian Divide.

The extremely high frequency of one allele within the

Great Lakes locations produced a small ®gr (<0.003)

between them, but differences between lake and riverine

populations led to a pooled ®gz = 0.10 across all regions.

Pairwise @sr values (Table 2) produced significant dif-

ferences between all lake samples and the two pooled

river groups, the Lake Erie tributaries and the Ohio River

tributaries, while the difference between the two river

groups was not significant even though private alleles

occurred in several rivers on each side of the divide

(Figure 2). The male-inherited form of CO1 showed more

sharing of haplotypes (Figure 3, accession numbers

MN125131-MN125143) yet they still indicated clear

separation between the Black and Cuyahoga rivers, from

where most samples derived.

We identified 4 different type A haplotypes among 9

individuals, 8 from the East Branch Black River and 1

from Eagle Creek, albeit a haplotype present in the Black

River (Figure 1). Combined with the Type A haplotypes

from Doucet-Beaupré et al. (2012), accession numbers

MN125127-MN125130, intraspecific variation of 9% at

the DNA base level occurs in P. grandis (Figure 4). All of

these individuals were morphologically ‘dlemetied! as P.

grandis, and sequences of male mitochondria of four of

the Black River mussels confirmed their identification.

These Type A sequences were similar in distance to the

divergence between P. grandis and the congeneric spe-

cies, P. cataracta and P. fragilis. Adding one type A

haplotype available from the Chottlnnadher River on the

Georgia/Alabama border (Genbank MG199625) and a

Type 'B haplotype from the Escambia River in southern

Alabama (Genbank MG199624), supported a hypothesis

of basal division between the two haplotype forms.

Furthermore, this phylogeny requires the divergence of

both forms prior to P. grandis migrating north.

R.A. Krebs et al., 2019 Page 77

Type B Haplotypes

Lake Erie (LE) H59

Niagara River Chagrin R ~ Ashtabula R group

LE River Tributaries H64Q group Huron R.

Y H60

ShioiN Thbutanes We 7 HS2 44,5 cf (~) H65_ Blanchard R group.

ses Lake Ontario 4... 9 9 H63 7? @ p? neo

; CXH53 ay nae

H54 yy Killbuck Ck group

H55_ :

Tinkers Ck H470 @ H70

H46 C— ® H71

H2 © & H72

H25 ;

OH17

H48 H29

H18

Additional unrelated single step Lake Erie variants

Figure 2. Haplotype network for a fragment of the COI gene amplified in P. grandis that were collected across the Lake Erie

watershed, Niagara River, Lake Ontario, many tributary streams of Lake Erie, and two streams below the northern divide in the Ohio

River watershed. The network reflects the Type B common female mitotype, to which the Type A variant connects distantly (Figure 4).

H35—45 occur west and northwest of the Great Lakes and therefore are not included here. Size of the circle reflects the number of that

haplotype found. Hatch marks indicate additional mutational steps and labels correspond to the name assigned in Genbank.

To address the process of divergence, Tajima’s D dif-

fered significantly (8, >> 9,,) in all lake samples (Table 1)

as previously reported (Krebs et al., 2015), but results for

rivers was confounded by the combined presence of the

Type A and Type B forms due to the large number of

segregating sites when Type A haplotypes are included. As

a consequence, Tajima’s D differed significantly in Lake

Erie tributaries only when the Type A form was excluded,

while the reverse occurred for the Ohio River tributaries.

A complementary test, mismatch analysis (in ARLEQUIN

Table 2. Population pairwise sr values among P. grandis individuals collected from the 5 regions. Samples derived from within Lake

Erie, the Niagara River, Lake Ontario, the Tributaries of Lake Erie, and two headwater tributaries of the Ohio River, Eagle Creek and

Kilbuck Creek in Ohio.

Region Lake Erie Niagara R. Lake Ontario Lake Erie Tributaries

Niagara R. 0.000

Lake Ontario 0.002 0.002

Lake Erie Tributaries 0.156*** 0.070*** 0.080***

Ohio R. tributaries 0.371*** 0.202*** 0.241 *** 0.011

Page 78

GM Lake Erie

[—_] Cuyahoga River

[__} Black River

[___] Other Rivers

MH12

MH8

MH6

THE NAUTILUS, Vol. 133, Nos. 3-4

MH13

MH1

MH10

Figure 3. Haplotype network for a fragment of the male-inherited COI gene amplified for a subset (N=28) of the P. grandis sampled

for population structure. Where multiple allele copies were found, samples came from both above and below the fall lines separating

upper and lower reaches. Size of the circle reflects haplotype sample number, with single copies found for all but three. Hatch marks

indicate additional mutational steps and labels correspond to the name assigned in Genbank.

and DnaSP), indicated no significant difference from pre-

dictions expected under population expansion (P >0.05)

in either tributary region or when pooling data for all of

the Lake Erie watershed (results not shown).

DISCUSSION

A star pattern of variation in P. grandis COI haplotypes

(Fig 2) was identified in each region, which infers that

one common haplotype is surrounded by many variants

of short branch lengths (Avise, 2000; Smietanka et al.,

2009). This pattern is characteristic of populations

founded by a small number of individuals that rapidly

expanded in size (Braverman et al., 1995). The pattern

has persisted even though populations today are much

reduced from the past, whether from the invasion of

dreissenid mussels depleting and restricting lake pop-

ulations to coastal areas (Zanatta et al., 2015; Bossenbroek

et al., 2018) or where urbanization has depleted habitat

in many rivers (Krebs et al., 2010a). Isolation played a

concurrent role, shown by a pattern of multiple allele

copies or short lineages of related sequences deriving

from a single stream.

The evolutionary model that best explains these hap-

lotype patterns in P. grandis is one of neutrality where

each mutation creates a new allele. Based on this neutral,

infinite-allele model, the observed variation in P. grandis

in Lake Erie suggests that the species exists as one almost

panmictic population (Krebs et al., 2015) from which the

species expanded into Lake Ontario as a recent coloni-

zation wave from a leading edge, which further reduced

variation. Today, one haplotype i in Lake Erie persists at an

allele frequency a little above 80%, rising to over 90% in

Lake Ontario. When P. grandis may Thaw entered Lake

Ontario is unknown. The. first Welland Canal connecting

lakes Erie and Ontario opened in 1829. But, P. grandis

also reached the lower Genesee and Oswego basins, the

lowland parts of the Mohawk River (Strayer and Jirka,

1997) and the Lake Champlain (Smith 1985) basins of

New York State perhaps through links to the Erie Canal

that connected the Cuyahoga and Allegheny Rivers,

among others (Strayer 1987, 1995). Canals would have

provided good habitat for lacustrine species like P. grandis

(Tevesz et al., 2002a).

Ortmann (1924) proposed that founder events from

the west likely produced the mussel assemblage in Lake

R.A. Krebs et al., 2019

Page 79

Red River watershed,

upper Minnesota

“ oN LER

EBRSTTS LOS

Ze, 2D * TITISAs

ED'S x= SP

%5,% 2

Pyganodon grandis

Type B

haplotypes

430 Pelican R., MN

Black River and

Eagle Ck

H75 Lake Erie

y7s Watershed, OH

47g RiceR. and Lake

St. Marie, MN

Pyganodon

grandis

ae TypeA

0.02 haplotypes

Figure 4. Gene tree for a fragment of the COI gene in P. grandis showing the distant relationship between the many Type B

haplotypes, and the Type A haplotypes, and haplotypes of two congeneric species that were similarly distant to each haplotype form.

Unless indicated, samples span the Midwest and Great Lakes regions. The southern variants fell at the base of each haplotype clade,

which supports a deep origin for both haplotype forms predating northern expansion. The Red River clade, which is the most divergent

group within the northern samples (Krebs et al., 2015), is highlighted as a contrast to Type A.

Erie, via the Wabash River and its connection to the

Maumee watershed, although Graf (2002) expanded the

model to multiple lake sources. Little discussion con-

sidered river origins for species like P. grandis. Variation

between Lake Erie populations and those of its

tributaries suggests either that mussels independently

reached the rivers or that additional colonization events

supplemented genetic variation derived from the lake.

Assemblages in the Cuyahoga and Black rivers changed

over time even before European settlements (Tevesz

et al., 2002b), and each river possessed markedly dif-

ferent groups above and below waterfalls that isolated

upper reaches (Dean, 1880; Lyons et al, 2007; Krebs

et al., 2010b). These rivers descend from a ridge line at

about 300 m down to Lake Erie at 175 m, and at least one

high waterfall formed in each river. In another locally

common mussel, Lampsilis siliquoidea (Barnes, 1823), the

impact of these falls among neighboring streams was visible

in large differences in female-inherited haplotype fre-

quencies, while the same male-inherited sequences could

be found above and below waterfalls (Krebs et al., 2013).

Movement across watersheds probably involved

headwater capture following glacial retreat and isostatic

rebounding, a process that is well documented across the

Great Lakes watershed subsequent to the last glaciers,

12000-15000 years ago (Coffey, 1958; Bishop, 1995). By

that time, populations of P. grandis had likely expanded in

the upper Ohio River south of the glacial line providing

more time for new mutations to arise from a common

haplotype. Simple mutation-drift dynamics could create

the presently observed pattems in genetic variation, re-

quiring only typical mitochondrial mutation rates (Haag-

Liautard et al., 2008: Hamilton, 2009) and modest his-

torical effective population sizes, which are a reasonable

expectation for this generalist species (Watters et al.,

2009). Partitioned sub-populations would enable drift to

randomly impact isolated alleles (Wright, 1931, 1932), and

do so differently among streams, to which additional mu-

tation and occasional migration can be added (Héssjer et al.,

2014). Thus, the post-glacial time period for the region likely

sufficed for both rare variants to arise and a common allele

to slowly decline following sequential colonization sweeps,

with related but different rare haplotypes arising in streams.

Genetically unusual if not unique for P. grandis is the

co-occurrence of two highly differentiated (9%) mtDNA

forms. Support for ancient divergence within P. grandis

over some form of introgression derives from the phy-

logeny (Fig 4.) that includes one Type A haplotype from

the Chattahoochee River on the Georgia/Alabama border

(Genbank Accession MG199625), and a Type B haplotype

came from the Escambia River in southern Alabama

(Genbank MG199624) (Smith et al., 2018 a, b). A BLAST

search showed each sequence to be only 96-97% similar

to their respective northern clade relatives, while all

northern sequences within both clades were much more

closely related to each other at 1-2%. Therefore, Type A

Page 80

and Type B mitochondrial types have likely co-occurred in

P. grandis a very long time, possibly since before the

radiation of the genus, and both haplotypes were carried

northward in migrants following glacial retreat. Other

haplotypes likely came too, and they reached across

headwaters where simple drift processes limited what

haplotypes remain today.

In sum, the historical movement of P. grandis dem-

onstrates how population structure may have arisen in

many mussels, arriving through different channels from

the south in the Pleistocene. Pyganodon grandis was just

more extreme in its success, as it reached across to eastern

Colorado (Liu et al., 1996), wp the Mississippi to northern

Minnesota (Krebs et al., 2015) and Canada (Doucet-

Beaupré et al., 2012), ail separately, along the Ohio River

and Wabash River to reach the lakes and through eastern

tributaries to access streams below the lakes. It has even

dispersed to the Escambia-Choctawhatchee watersheds

(Haag, 2012) and south to Mesoamerica (Pfeiffer et al.,

2019), probably long before any colonization of the Great

Lakes. Perhaps these areas were reached when sea levels

were low and glaciers high (Swift et al., 1986). Yet, the large

lacustrine populations heme the least genetically diverse,

probably from rapid and sequential population expansion, a

process that favors few or one allele compared to the ex-

pectations with stream capture of already diverse populations.

ACKNOWLEDGMENTS

Collections in the Great Lakes were funded by the U.S.

Fish and Wildlife Service — Great Lakes Fish and Wildlife

Restoration Act (#30181AG152). Other financial support

included a Research Experiences for Undergraduates

award from the National Science Foundation (DBI

0243878), and M. Lyons, who initiated the tributary work,

was supported by a National Oceanic and Atmospheric

Administration fellowship (NA0O7-NOS4200018), while

RAK was supported by an Established Full-time Faculty

Research and Dev ‘elopment award from Cleveland State

University. Scientific collection permits were provided by

wildlife agencies of Ohio, New York, and Ontario. Many

helped provide specimens for this study over the years,

A. Karatayev, B. Tulumello and K. Bauer (SUNY Buffalo

State College) plus Wendy Paterson, Traci Griffith, Mariah

Scott, Ethan Nederhoed, Lindsey Kolich, and Emily

Marlow (CMU) all assisted in sampling i in New York, while

Paul Doerder, Louie Rundo, Dan Gouch, Mark Lyons,

Jeremy Holt, John Hook, David Kriska, Jeff Pallotta, and

Erin Steiner helped collect in various tributary streams, and

Mark Lyons, Matt Begley and Nikko Hogya helped with

DNA processing. We thank John Pfeiffer and Kevin Cum-

mings for many constructive suggestions during review. This

article is contribution #132 of the Central Michigan Uni-

versity Institute for Great Lakes Research.

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Appendix 1. Supplemental Table. The samples used in the present study labeled by haplotype number. The main biogeographic

region is provided relevant to previous work (Krebs et al., 2015) with new samples for Lake Ontario, Lake Erie (LE) tributary rivers and

two rivers from south of the Laurentian Divide, Kilbuck and Eagle Creeks, which are in the Ohio River watershed. Subwatersheds list

the specific area of collection along with sample size (N), Genbank accession numbers, and a pair of coordinates within the range of that

haplotype.

Haplo-

ee Region Sub watershed N_ Accession # Latitude Longitude

H1 All regions except Red River most if not all sub-watersheds 416 KM262507.1

H2 LE & Tributary Sheldon Marsh, Vermilion River 2 KM262508.1 41.4246 -82.6242

H3 Lake Erie Sheldon Marsh 1 KM262509.1 41.4246 -82.6242

H4 LE & Sandusky Bay Old Woman Creek, Yellow Swale 2 KM262510.1 41.3794 -82.5117

H5 Lake Erie Old Woman Creek 1 KM262511.1 41.3794 -82.51173

H6 LE, Niagara R. & Tributaries Many nearshore sites & Ashtabula River 8 KM262512.1 41.8582 -—80.6397

H7 Sandusky Bay South Creek 1 KM262513.1 41.398 -—83.0106

Hs Sandusky Bay South Creek, Yellow Swale 2 KM262514.1 41.398 -83.0106

H9 Sandusky Bay Muddy Creek Bay 1 KM262515.1 41.4496 —S3.0281

H10 Lake Erie Plum Brook 1 KM262516.1 41.4247 -82.639

Hill Lake Erie Cranberry Creek 1 KM262517.1 41.3824 -82.4727

H12 Lake Erie Cranberry Creek 1 KM262518.1 41.3824 -82.4727

H13 Lake Erie Crane Creek 2 KM262519.1 41.6275 -—83.2018

H14 Lake Erie & Sandusky Bay Turtle Creek 3 KM262520.1 41.6031 -83.1517

H15 Lake Erie Turtle Creek 2 KM262521.1 41.6031 -83.1517

H16 Sandusky Bay & Niagara R. Grand Isle & Muddy Creek Bay 2 KM262522.1 43.0395 —78.8937

H17 Niagara River & Lake Grand Isle & Eastern inlets 3 KM262523.1 43.6254 —76.1959

Ontario

H18 Lake Erie Misery Bay 1 KM262524.1 42.1614 -80.0898

H19 Lake Erie Duck Pond 1 KM262525.1 42.0956 -80.071

H20 Lake Erie Misery Bay 1 KM262526.1 42.1614 -80.0898

H21 Lakes Erie & St Clair East Harbor, Young Marsh 3 KM262527.1 41.5421 -82.8104

H22 Lake Erie Crane Creek 1 KM262528.1 41.6275 -83.2017

H23 Sandusky Bay Muddy Creek Bay 1 KM262529.1 41.4496 -83.0281

H24 Sandusky Bay Muddy Creek Bay 1 KM262530.1 41.4496 -83.0281

H25 Lake Erie East Harbor 1 KM262531.1 41.5421 -82.8104

H26 Lake Erie North Maumee Bay, Turtle Creek 2 KM262532.1 45.9184 -89.5324

H27 Lake Erie Toussiant Creek 2 KM262533.1 41.5784 -83.1085

H28 Lake Erie and Niagara R. Turtle Creek, Strawberry Island 2 KM262534.1 41.6031 -83.1517

H29 Lake Erie Turtle Creek 1 KM262535.1 41.6031 -83.1517

H30 Lake Erie Turtle Creek 1 KM262536.1 41.6031 -83.1517

H31 Niagara River Strawberry Island 1 KM262537.1 42.9539 —78.9235

H32 Niagara River Strawberry Island 1 KM262538.1 42.9539 —-78.9235

H33 Niagara River Strawberry Island 1 KM262539.1 42.9539 —78.9235

H34 Niagara River Strawberry Island 1 KM262540.1 42.9539 —78.9235

H35 Minnesota (NW) Red River and upper lakes, Minnesota. 8 KM262541.1 47.8253 -—93.3748

H36 Minnesota Lac Qui R.; lower Minn R., Minn. 3 KM262542.1 45.0166 —95.8866

H37 Minnesota (NW) Rice R., Deer L. Upper Minnesota 6 KM262543.1 46.5326 —93.32

H38 Minnesota (NW) Pfeiffer Lake, Upper Minnesota 4 KM262544.1 47.7515 -92.4771

H39 Minnesota (NW) Pfeiffer Lake, Upper Minnesota 1 KM262545.1 47.8253 —93.3748

H40 Minnesota (NW) Prairie River, Upper Minnesota 12 KM262546.1 47.2391 -—93.4821

H41 Minnesota (NW) Prairie River, Upper Minnesota 1 KM262547.1 47.2391 —93.4821

H42 Minnesota (NW) Prairie Lake, Michigan 2 KM262548.1 41.8586 —-85.4037

H43 Minnesota (NW) Big Fork R., Upper Minnesota 1 KM262549.1 47.8004 -—93.5724

H44 Upper Great Lakes North Twin Lake, WI 1 KM262550.1 46.0665 -—89.0887

H45 Upper Great Lakes Annabelle & St. Germaine Lakes, WI 2 KM262551.1 46.2206 -—89.6787

H46 LE Tributary Black, Grand, Cuyahoga (above falls) Rivers 3 MN125095 41.4545 -82.1354

H47 LE Tributary Cuyahoga River (Tinkers Creek, above falls) 3 MN125096 41.3146 -81.4354

H48 LE Tributary Cuyahoga River (Tinkers Creek, above falls) 1 MN125097 41.3146 —-81.4354

H49 LE Tributary Cuyahoga River (Tinkers Creek, above falls) 1 MN125098 41.3146 —-81.4354

H50 LE Tributary Cuyahoga River (Tinkers Creek, above falls) 2 MN125099 41.3146 -81.4354

H51 LE Tributary Black River 1 MN125100 41.4545 -82.1354

H52 LE Tributary Black River 1 MN125101 41.4545 82.1354

H53 LE Tributary Black River, Cuyahoga River (above falls) 29 MNI125102 41.4545 -82.1354

(Continued)

THE NAUTILUS

, Vol. 133,

Sub watershed

Accession #

Latitude

Nos. 3-4

Longitude

Black River West Branch

Page 84

Appendix 1. (Continued)

Haplo-

type Region

H54 LE Tributary

H55 LE Tributary

H56 LE Tributary

H57 LE Tributary

H58 LE Tributary

H59 LE Tributary

H60 LE Tributary

H61 LE Tributary

H62 LE Tributary

H63 LE Tributary

H64 LE Tributary

H65 LE Tributary

H66 LE Tributary

H67 Ohio River Tributary

H68 Ohio River Tributary

H69 Ohio River Tributary

H70 Ohio River Tributary

H71 Lake Ontario

H72 Lake Ontario

H73 LE Tributary

H74 LE Tributary

H75 LE Tributary

H76 LE & Ohio R. Tributary

H78 Quebec, Canada

H79 Quebec, Canada

H80 Minnesota (NW)

HSs1 Minnesota (NW)

MH1 LE Tributary

MH2 LE Tributary

MH3 LE Tributaries

MH4 LE Tributary

MH5 LE Tributary

MH6 LE Tributary

MH7 LE Tributaries

MHS8 LE Tributary

MH9 LE Tributary

MH10 LE Tributary

MH11 LE Tributary

MH12 LE Tributary

MH13 Lake Erie

Cuyahoga River (above falls)

Ashtabula River East Branch

Ashtabula River West Branch

Huron River West Branch

Rocky River East Branch (drained pond)

Chagrin River (above falls)

Blanchard River

Killbuck Creek

Eagle Creek (Mahoning R. watershed)

Blind Sodus Bay

North Bay

Black River East Branch

Black River East Branch, Eagle Creek

Lac Sainte Marie

Pelican Lake Minnesota

Rice River

Cuyahoga River (above falls)

Black River

Black River West Branch, Tinkers Creek,

Ashtabula River

Cuyahoga River (Tinkers Creek, above falls)

Black River

Conneaut Creek

Diverse Rivers, Sandusky Bay

Black River East Branch

Huron River west branch

Cuyahoga River & Tinkers Creek (above falls)

Black River East Branch

Sandusky Bay

CO OD ND EB ODD FD OB ET

SSS SSS SS SS SS< SSS 5S <SSSS5S SS 5S SS SS SS 55SS8s5

FD a

N125103

N125104

N125105

N125106

N125107

N125108

N125109

N125110

N125111

N125112

N125113

N125114

N125115

N125116

N125117

N125118

N125119

N125120

N125121

N125122

N125123

N125124

N125125

N125127

N125128

N125129

N125130

N125131

N125132

N125133

N125134

N125135

N125136

N125137

N125138

N125139

N125140

N125141

N125142

N125143

41.3141

41.3018

41.8119

41.7645

41.286

41.2445

41.5257

40.8915

40.9528

41.2827

43.338

43.6314

41.2363

45.9561

48.0633

46.5326

41.3018

4145453

41.2951

41.3146

41.45453

41.9041

41.4195

41.2363

41.286

41.3018

41.2363

41.4195

—§2.1316

—81.2026

—80.5973

80.5973

—80.6155

—§2.6435

81.6779

—§1.6779

—81.2605

81.2605

83.5643

—82.0262

—§2.0262

82.0262

—§1.118

—76.7281

—76.1919

82.0797

—82.0797

82.0797

—75.9305

—92.8321

—93.32

—81.2026

—§2.1354

82.1456

—§1.4354

82.1354

—§0.5289

—82.9227

—82.0797

82.6435

—82.6435

—8 1.2026

—82.0797

—82.9227

THE NAUTILUS 133(3-4):85-93, 2019

Page 85

Description of two new muricid species (Gastropoda: Muricidae:

Muricopsinae) from the western Atlantic and the eastern Pacific

Roland Houart

Institut royal des Sciences naturelles de Belgique’

and

Muséum national @’Histoire naturelle, Paris, France”

UMR7205 ISyEB

roland. [email protected]

Ne sd. | .———iiill ll ———————————s

ABSTRACT

Two new species of Muricidae are described from Brazil and

from the Pacific coast of Panama. Favartia aquinoi new species

from Brazil is compared with the related F. glypta (M. Smith,

1938), as well as with F. cellulosa (Conrad, 1846) and F. levicula

(Dall, 1889). Muricopsis vassarti new species is described from

the Pacific coast of Panama and compared with three other

Muricopsis species from the Eastern Pacific, M. zeteki Hertlein

and Strong, 1951, M. westonensis Myers and D’Attilio, 1990 and

M. taupini Garrigues, 2016.

Additional Keywords: Favartia, Muricopsis new species, Brazil,

Panama, eastern Pacific

eee EST

INTRODUCTION

Muricopsinae comprise 13 genera and one subgenus

[Muricopsis (Risomurex)| and include some 268 species

(Houart, 2018, updated). Two new species are described

herein, the first, from Brazil, is assigned to Favartia

Jousseaume, 1880, the second is a Muricopsis s.s. species

from Pacific Panama.

Thirty Recent species are assigned to the genus

Favartia in the Western Atlantic (MolluscaBase 2019a).

This number includes species of Favartia (M urexiella) and

may slightly differ in recent publications, depending on

what synonyms are considered by the authors.

A group of species is of particular interest. It is com-

posed of F. glypta (M. Smith, 1938), F. cellulosa (Conrad,

1846), F. levicula (Dall, 1889) and a new species described

from Brazil.

Rios (1985) considered Murexiella iemanja Petuch,

1979, described from the Abrolhos Archipelago in

Brazil, a synonym of F. glypta from the Pliocene of

Clewiston, Florida, but also known from the Recent

fauna and occurring in several places from Florida to

1 ‘

Research Associate

% 3

Research Associate

Brazil. He was followed by Houart (1991: 32) and by

Vokes (1994: 112).

Favartia glypta was considered a synonym of F. levicula

by Radwin and D’Attilio (1976: 159). However, these two

species differ in shell oramentation and protoconch

characters. The protoconch of F. glypta is paucispiral,

consisting of 1.5 rounded whorls (Figures 3, 25) while that

of F. levicula is conical and multispiral, consisting of al-

most 4 whorls (Figure 37).

The Western Atlantic and Eastern Pacific species

assigned to Muricopsis by several authors, such as Keen

(1971), Vokes (1971, 1994), Kaicher (1974, 1978, 1980,

1991), Fair (1976), Radwin and D’Attilio (1976) and many

others are now assigned to Muricopsis and to Murexsul

(MolluscaBase, 2019b), depending on their shell mor-

phology (Merle and Houart, 2003).

The species of Muricopsis are characterized by the

hypertrophy of apertural denticle D2 and by strong col-

umellar folds. The spiral sculpture consists of primary

cords P1—P5 on the convex part of the teleoconch whorl,

atrophied P6 on the siphonal canal, followed by primary

cords ADP and occasionally MP. In Murexsul oxytata (M.

Smith, 1938) and other species of that group, the P2

primary cord is atrophied and the columellar folds are

absent.

Of the four Eastern Pacific species that had been

included in Muricopsis, two remain in Muricopsis: M.

pauxilla (A. Adams, 1854) and M. zeteki Hertlein and

Strong, 1951, and two have been reassigned to Murexsul:

M. armatus (A. Adams, 1854) and M. jaliscoensis

(Radwin and D’Attilio, 1970). While the spiral sculpture

of M. pauxilla is typical for Muricopsis, it lacks colu-

mellar folds. Two additional species of Muricopsis were

subsequently described, M. westonensis Myers and

D Attilio, 1990, from Cocos Island, Costa Rica and M.

taupini Garrigues, 2016, from the Galapagos. A fifth

species of Muricopsis, syntopic with M. zeteki, is here

described as new.

A broad phylogenetic analysis of Muricopsinae is

needed in order to verify the status of some genera and

assigned species.

Page 86 THE NAUTILUS, Vol. 133, Nos. 3-4

Number of whorls (here 2 1/4)

WBiey winuixeyy

Figures 1-6. Two new species of Muricopsinae. 1. Method for determining diameter, height and counting the number of protoconch

whorls. 2-5. Favartia glypta (M. Smith, 1938). 2. Paratype of Murexiella iemanja Petuch, 1979. Brazil, Bahia State, 2 km E Santa

Barbara Island, Abrolhos Archipelago, Chapeirao Reef, 25 m, USNM 780653. 3. Protoconch of Murexiella iemanja Petuch, 1979,

holotype USNM 780652. 4-5. Florida, Hendry County, Clewiston, Caloosahatchee Formation, Pliocene, Holotype, University of

Alabama, Museum of Natural History, 25.3 mm (reproduced from Vokes, 1968). 6. Distribution of Favartia aquinoi new species (red

stars) and Muricopsis (Muricopsis) vassarti new species (red square). Scale bar = 500 wm.

R. Houart, 2019

Page 87

MATERIALS AND METHODS

The new material studied was collected between 1960 and

2008 in Pacific Panama and along the Brazilian coast. The

material used for comparison is deposited in MNHN,

USNM and in the author’s private collection.

Characters used to describe shell morphology address

the general aspect of the shell, its shape, size, and color,

the shape of the spire including the protoconch, the

number and features of the teleoconch whorls, details of

the suture and of the subsutural ramp, details of axial and

spiral sculpture, the aperture, the siphonal canal and the

operculum. The description is based on the type material.

The method used to determine diameter and height,

and to count the number of protoconch whorls, follows

Bouchet and Kantor (2004) as shown in Figure 1.

Abbreviations used in the text are as follows: IRSNB:

Institut royal des Sciences naturelles de Belgique, Bruxelles,

Belgium; MNHN: Muséum national d Histoire naturelle,

Paris, France: RH: R. Houart Collection; USNM: National

Museum of Natural History, Washington, DC, USA; ad:

adult; dd: collected empty; juv: juvenile; lv: collected alive.

Terminology used to describe the spiral cords and the

apertural denticles (after Merle 2001, 2005) (Figures 7, 8,

24, 27, 40, 41). Terminology in parentheses: variable feature.

Convex part of teleoconch whorl and siphonal canal:

Adis: Adapical infrasutural secondary cord on subsutural

ramp; IP: Infrasutural primary cord on subsutural ramp;

P1—P6: Primary spiral cords on the convex part of the

teleoconch whorl and the siphonal canal; s1—s5: Secondary

cord of the convex part of the teleoconch whorl (for

example, sl: Secondary cord of the convex part of the

teleoconch whorl between P1 and P2); ADP: Adapertural

primary cord on the siphonal canal; MP: Median primary

cord on the siphonal canal; ABP: Abapertural primary

cord on the siphonal canal.

Aperture: ID: infrasutural denticle; D1 to D6: abapical

denticles.

SYSTEMATICS

Family Muricidae Rafinesque, 1815

Subfamily Muricopsinae Radwin and D’Attilio, 1971

Genus Favartia Jousseaume, 1880

Type Species: Murex breviculus Sowerby II, 1834, Indo-

West Pacific, by original designation.

Favartia aquinoi new species

Figures 6, 7-19

Type Material: Holotype IRSNB MT.3801/1.G.34044, 1

paratype coll. Wanderley Vieira de Aquino Junior, Sao

Paulo, Brazil; 1 paratype R. Houart, all from the type

locality.

Type Locality: Brazil, Rio de Janeiro State, off Arraial do

Cabo, 30-35 m, 2008.

Other Material Examined: Brazil, Ilha do Pai, 16 m,

1960, RH (1 dd, ad); Ilha de Sao Sebastiao, 35-45 m, in

sand, 1992. RH (2 lv, ad): Vitoria Bank, 52 m, 1987, RH

(1 dd, juv).

Distribution: Brazil, from Vit6ria Bank to Ilha de Sao

Sebastiao, living at 35-45 m.

Description: Shell medium sized for the genus, up to

17.9 mm in length. Height/width ratio 1.9-2.0. Slender,

lanceolate, biconical, broadly ovate. Very weakly spinose,

nodose, lightly built. Subsutural ramp moderately broad,

weakly sloping, weakly concave. Shell color creamy white

or light tan with occasional brown spots on subsutural

ramp, between axial varices. Aperture pale white within.

Spire high, with 1.5 protoconch whorls and up to 5

broad, convex, strongly shouldered teleoconch whorls.

Suture impressed. Protoconch large, broad. Whorls

rounded. Maximum width and height 900 pm. Axial

sculpture of teleoconch whorls consisting of low, narrow,

weakly frondose varices. Each varix with very short,

frondose, open, primary spines. First teleoconch whorl

with 8 varices, second with 8 or 9, third to penultimate

whorl with 9, last whorl with 7 or 8 varices. Spiral

sculpture of high, rounded, narrow, weakly nodose pri-

mary and secondary cords consisting of (adis), IP, Pl—

ABP. Adis and IP shallow, giving rise to a small, curved,

short spine at intersection of axial varices. P1—P5 of same

height and wide on last teleoconch whorl; P6 quite nar-

rower and lower, followed by ADP, MP, and ABP, ADP,

and MP of same strength as PI—P5; ABP somewhat

smaller. Low, blunt, open spines originate at crossing of

axial varices and spiral cords, more apparent on apertural

varix. Aperture moderately large, ovate. Columellar lip

narrow, smooth. Rim partially erect, adherent at small

portion at adapical extremity. Anal notch shallow, broad.

Outer lip erect, crenulated, with low ID and D1-D6

within. Siphonal canal short, 18-21% of total shell length,

narrow, weakly dorsally recurved, narrowly open, with

short, blunt spines corresponding to ADP, MP, and ABP,

decreasing in length abapically. Operculum and radula

unknown.

Remarks: Favartia aquinoi new species may be com-

pared with F. glypta (= Murexiella iemanja) which also

occurs in Brazil, with F. cellulosa, living from Florida to

Brazil, and with F. levicula known from North Carolina to

the Gulf of Mexico. The most similar species, F. glypta

(Figures 2-5, 20-23, 24-31), differs from F. aquinoi

new species in having a somewhat smaller shell in

Recent specimens with a smaller protoconch, a lower

spire, narrower and more squamous primary spiral

cords, more distant from each other, a very narrow s2

cord and a narrower siphonal canal with short ADP,

MP, and occasionally ABP, spines. These differences

can already be observed in juvenile specimens of both

species, for example in the holotype of Murexiella

iemanja (8 mm) (Figures 20-21) anda young F. aquinoi

new species (5.4 mm) (Figures 17-19). Favartia cel-

lulosa (Figures 32-34), mostly known from Florida, but

Page 88 THE NAUTILUS, Vol. 133, Nos. 3-4

Figures 7-23. Favartia species. 7-19. Favartia aquinoi new species Brazil, Rio de Janeiro State, off Arraial do Cabo, 30-35 m,

sand and rubble, Sept. 2008. 7-11. Holotype MT.3801/1.G.34044, 17.8 mm. 12-14. Paratype RH, 17.9 mm.15. Paratype coll.

Wanderley Vieira de Aquino Junior, 16.4 mm. 16. Protoconch (paratype RH), crushed afterwards. 17-19. Brazil, Vitoria Bank, 20°32’ S,

38°11 W, 52 m, RH, 5.3 mm. 20-23. Favartia glypta (M. Smith, 1938). 20-21. Brazil, Bahia State, 2 km E Santa Barbara Id., Abrolhos

Archipelago, Chapeirao reef, 25 m, holotype of Murexiella iemanja Petuch, 1979, USNM 780652, 7.8 mm (photo courtesy USNM).

22-23. Brazil, Espirito Santo State, off Guarapari, 45-60 m, June 2012, RH, 10.6 mm. Scale bars (also for Figure 25) = 500 pm.

R. Houart, 2019 Page

Figures 24-37. Favartia species. 24-31. Favartia glypta (M. Smith, 1938). 24-25. Brazil, Espirito Santo State, off Guarapari, 45-60

m, June 2012, RH, 10.6 mm. 26-27. Colombia, southern Cartagena, off Golfo de Morrasquillo, 100 m, March 2005, RH, 13.9 mm.

28-30. Brazil, Espirito Santo State, off Guarapari, 25m, Nov. 1993, RH, 11.1 mm. 31. Brazil, Espirito Santo State, Gurapari Channel,

muddy area, low tide, April 1995, RH, 15.4 mm. 32-34. Favartia cellulosa (Conrad, 1846). Florida, West of Boca Raton, 54 m, rubble

bottom. RH. 16.2 mm. 35-37. Favartia levicula (Dall, 1889). 35-36. Florida (no other data), RH, 15.5 mm. 37. Florida, Southwest of

Panama City, dredged 55 m, protoconch, RH. Scale bar = 500 pm.

Page 90 TMUILOIS, Wall, 188. IN@s, G4

Figures 38-49.

Muricopsis species. 38-44. Muricopsis (Muricopsis) vassarti new species. 38-41, 43. Pacific Panama,

Archipiélago de las Perlas, 1997, holotype MNHN-IM-2000-305226, 22.8 mm. 42. Pacific Panama, Archipiélago de las Perlas, Isla Pedro

Gonzales, 8° N, 79° W, 2002, paratype RH, 23.4 mm. 44. Pacific Panama, Archipiélago de las Perlas, Isla Pedro Gonzales, 8° N, 79° W,

2002, paratype RH, 18.9 mm. 45-49. Muricopsis (Muricopsis) zeteki Hertlein and Strong, 1951. 45-47. Pacific Panama, Archipiélago

de las Perlas, 1997, RH, 21.0 mm. 48. Pacific Panama, Archipiélago de las Perlas, Isla Pedro Gonzales, 8° N, 79° W, 2002, RH, 28.3 mm.

49. Protoconch, Pacific Panama, Archipiélago de las Perlas, Isla Pedro Gonzales, 8° N, 79° W, 2002, RH. Scale bar = 500 wm.

R. Houart, 2019

Page 91]

Figures 50-53. Muricopsis species. 50-51. Muricopsis (Muricopsis ) westonensis Myers and D’Attilio, 1990, Costa Rica, Cocos Id,

Bahia Weston, 12 m, under dead coral, April 1983, holotype USNM 860014, 13.4 mm (photo courtesy USNM). 52-53. Muricopsis

(Muricopsis) taupini Garrigues, 2016. Galapagos, off Santiago Ids, Albany Id, 5-15 m, on rocks, holotype MNHN-IM-2000-31680,

19.9 mm (photo Manuel Caballer, MNHN).

extending its geographical distribution to Brazil, differs in

having a broader shell, a smaller protoconch, comparatively

narrower primary cords, almost obsolete P6 cord and spine,

shorter or almost obsolete, broad, blunt P1—P6 spines,

broader and fewer axial varices, and a broader siphonal

canal, strongly tapered abapically, with a short ADP spine.

Favartia levicula (Figures 35-37) differs in having a

broader shell with a lower spire, broader and flatter primary

spiral cords, obsolete or strongly reduced between each

pair of axial varices, a shorter siphonal canal, strongly

adapically tapered, and chiefly in having a conical proto-

conch of 3.5 whorls, denoting planktotrophic larval de-

velopment (Figure 37), as opposed to a broad, globose,

paucispiral protoconch in F. aquinoi new species (Figure 16).

Favartia glypta was considered a junior synonym of F. levicula

by Radwin and D’Attilio (1976: 159), but this was certainly

due to Clench and Pérez-Farfante (1945: 56, pl. 28, figs 1-3),

who illustrated two specimens of F. glypta as Murex cellulosus

leviculus, following in that the taxonomy of M. Smith (1939:

16, pl. 13, fig. 8).

Etymology: I am pleased to follow the request of Marcus

Coltro, naming this new species after W. anderley Vieira de

Aquino Junior, shell collector from Sao Paulo and past

president of Conquiliologistas do Brasil.

Genus Muricopsis Bucquoy and Dautzenberg, 1882

Subgenus Muricopsis Bucquoy and Dautzenberg, 1882

Type Species: Murex blainvillei Payraudeau, 1826

(= Murex cristatus Brocchi, 1814), Mediterranean, by

original designation.

Muricopsis (Muricopsis) vassarti new species

Figures 6, 38-44

Type Material: Holotype, MNHN-IM-2000-305226,

Pacific Panama, Archipiélago de las Perlas, 1997; 3 par-

atypes, Pacific Panama, Archipiélago de las Perlas, Isla

Pedro Gonzales, 8° N, 79° W, 2002, R. Houart.

Type Locality: Pacific Panama, Archipiélago de las

Perlas.

Distribution: Pacific Panama, Archipiélago de las Perlas.

Description: Shell medium-sized for the genus, up to

23.4 mm in length. Height/width ratio 1.6. Lanceolate,

biconical, broad. Heavy and spinose. Subsutural ramp

broad, strongly sloping, strongly concave. Light brown

with darker brown spots on axial varices, between and on

spines. Creamy white on dorsal side of spines. Aperture

glossy white. Spire high, acute. Protoconch whorls eroded.

Teleoconch of up to 8 angulate, strongly shouldered,

spinose whorls. Suture weakly adpressed.

Axial sculpture of teleoconch whorls consisting of

narrow, high ribs on first teleoconch whorls and high,

broad, sharp spinose varices from fourth to last whorl.

First to penultimate whorl with 7 or 8 ribs or varices. Last

whorl with 6 varices. Spiral sculpture of low, strong,

narrow, squamous primary and secondary cords and few

threads. Last teleoconch whorl with broad IP, occasional

abis, followed by P1, P2, s2, P3, s3, P4, s4, P5, (s5), P6,

ADP. Primary cords giving rise to short, acute, tri-

angular, open spines on axial varices. Aperture large,

ovate. Columellar lip moderately broad, weakly flaring,

with 2 elongate, strong folds abapically. Rim partially

Page 92

erect, adherent at a small portion at adapical extremity.

Very low parietal tooth at adapical extremity. Anal notch

deep, narrow. Outer lip weakly erect, crenulated, with 5

strong denticles within, decreasing in strength abapi-

cally, consisting of IP, D1—D4. Siphonal canal short,

11-15% of total shell length, broad, weakly dorsally

recurved, open, with Ante, short P6 and ADP spines.

Operculum dark brown, strongly ovate with apical nu-

cleus in lower right. Radula unknown.

Remarks: The four studied specimens of Muricopsis

vassarti new species were collected living in syntopy

with two lots of M. zeteki, the latter being obviously

more numerous. One specimen, the holotype of M.

vassarti new species and 12 specimens of M. zeteki

were taken in one spot, three specimens (paratypes

RH) were collected in the other spot, together with 9

specimens of M. zeteki. Both lots were left unsepar ated

during many years, until a recent short review of the

Muricopsis and Murexsul species in my collection.

Muricopsis zeteki (Figures 45-49) differs from the new

species in having a Iexalleislh brown and white coloured

shell and a different spiral sculpture. In M. zeteki there

are 4-6 narrow spiral threads between each pair of

primary cords (Figures 48) as opposed to a single small

secondary cord viel one or two additional thea i in M.

vassarti new species (Figure 43). In M. zeteki the

primary cords are also narrower. The protoconch of M.

vassarti new species was eroded in all examined

specimens but could be paucispiral, consisting of 1.25

whorls (see below). It is conical and multispiral, con-

sisting of 3 whorls in M. zeteki (Figure 49) denoting

planktotrophic larval development, sibiglh may explain

its extended geographical distribution from the

northern Gulf of California to North Peru and the

Galapagos Islands. Garrigues (2016: 8) in his de-

scription of M. taupini described the protoconch of M.

zeteki as being paucispiral, consisting of 1.25 whorls.

The illustrated protoconch (Garrigues, 2016, figs 3I-])

is from a juvenile specimen collected at Boyerana Id, in

the Archipiélago de las Perlas and shows indeed a

paucispiral protoconch. However, the shell (fig. 31)

looks much closer to M. vassarti new species and could

be a juvenile specimen of that species. Muricopsis

westonensis from the Pacific coast of Costa Rica

(Figures 50-51) differs from Muricopsis vassarti new

species in having a smaller, narrower shell with a

somewhat less acute spire, a narrower aperture and

stronger folds on the columellar lip. It is also less spiny

with roe, brown coloured knobs at intersection of spiral

and axial sculpture instead of sharp, whitish spines in

M. vassarti new species. Finally, Muricopsis taupini from

the Galapagos (Figures 52-53) is different in having a

smaller shell with a lower spire, lower columellar folds tl

apertural denticles, a shorter siphonal canal and blunt,

blackish brown knobs.

Etymology: Named for André Vassart who collected

these specimens in 1997 and 2002, traveling aboard his

vessel LA BOUDEUSE.

THE NAUTILUS, Vol. 133, Nos. 3-4

ACKNOWLEDGMENTS

I am very grateful to Jose and Marcus Coltro, Brazil,

who sent me the Favartia specimens for study a few

years ago, to Ellen Strong, National Museum of Natural

History, Washington, DC, USA, for the permission she

gave to use the images from USNM, to Manuel Caballer

(MNHN) who provided the images of the MNHN

holotype of Muricopsis taupini (E-Recolnat Project:

ANR-11-INBS-0004), to Emily H. Vokes, Ponchatoula,

Louisiana, USA, for permission to reproduce her photo

of the holotype of Favartia glypta, and as always, to John

Wolff, Lancaster, Philadelphia, USA, for checking the

English text and for other comments. Many Thevales also

to ‘ihe reviewers, Yuri Kantor and Jerry Harasewych for

their useful advice.

LITERATURE CITED

Bouchet, P. and Yu.I. Kantor. 2004. New Caledonia: the major

center of biodiversity for volutomitrid mollusks (Mollusca:

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THE NAUTILUS 133(3-4):94-101, 2019

Page 94

Description of two new species (Bivalvia: Vesicomyidae,

Verticordiidae) from a cold seep in the South China Sea

Jinxiang Jiang

Third Institute of

Yaqin Huang

Third Institute of Oceanography

Oceanography Ministry of Natural Resources

Ministry of Natural Xiamen, 361005, CHINA

Resources and

Xiamen, Fisheries College

361005, CHINA Jimei University

Xiamen, 361021, CHINA

Qianyong Liang

MLR Key Laboratory of Marine

Mineral Resources

Guangzhou Marine

Geological Survey and

China ‘Geological Survey

Guangzhou, 510070, CHINA

Junlong Zhang’

Institute of Oceanology

Chinese Academy of istiencce

Qingdao, 266071, CHINA

Center for Ocean Mega-Science

Chinese Academy of Sciences

Qingdao, 266071, CHINA

ABSTRACT

In this paper, we describe two new deep-sea bivalve species

belonging to the families Vesicomyidae and Verticordiidae, re-

spectively. They were collected during a survey of the Haima

Methane Seep at the Qiongdongnan Basin on the northwestern

slope of the South China Sea, China, at a depth of 1,400 m.

Vesicomya rhombica new species is small in size and charac-

terized by its rhombic shell shape and a slight furrow on the

surface of the posterior shell region, running Rion the umbo to

the postero-vental margin. It is the first species of this genus

recorded from Chinese waters. Spinosipella xui new species has

a long, lamellar posterior lateral tooth on the right valve, 13

strong radial ribs, and conspicuous spines on the shell surface.

The latter constitutes one of three species of this genus living in

Chinese waters.

Additional Keywords: new species, Vesicomya, Spinosipella,

Vesicomyidae, Verticordiidae, taxonomy, chemosymbiotic

environment

INTRODUCTION

Many habitats remain underexplored in the bathyal and

abyssal seafloor that draw significant concerns worldwide.

Our knowledge of the biodiversity of deep-sea mollusks

remains quite limited due to technological difficulties in

the access to this perpetually dark world and resulting

insufficient sampling, which results in controversial ex-

planations of their fauna, biodiversity, and biogeogr aphy

(Arbizu and Brix, 2008: Brékeland ‘and George, 2009).

With the gradual increase in exploration of the deep-sea,

vast numbers of mollusks have been sampled. Reporting

newly found species is imperative to understanding the

biodiv ersity of the deep-sea, as accurate ‘dlemalReattone

and taxonomy are crucial and the foundation for further

studies.

' Corresponding author: [email protected]

The Vesicomyidae Dall and Simpson, 1901 is a deep-

sea bivalve family, distributed worldwide from about

100 m to more than 10,000 m depth in the abyssal and

hadal zones (Krylova et al., 2018). It has received mount-

ing attention due to its specialized habitat. Most of the

members are confined to chemosynthetic communities

such as cold seeps, hydrothermal vents, whale carcasses, or

other sulphide-rich reducing environments (Krylova and

Sahling, 2010; Coan and Welleratiielhe Scott, 2012; Krylova

et al., "2018). This family is considered to saralknrdls two

subfamilies, Vesicomyinae Dall and Simpson, 1901 and

Pliocardiinae W ‘oodring, 1925, totaling more than 125

Recent species (Krylova and Sahling, 2010; Decker et al.,

2012: Johnson et al., 2017; Krylova et al., 2018). These

two subfamilies are well- -supported and widely accepted

(Krylova and Sahling, 2010; Valdés et al., 2012). Ves-

icomya Dall, 1886 is the only genus of Vesicomyinae;

included species are generally small in size and mainly

inhabit abyssal plains or hadal trenches. Krylova and

Sahling (2010) assigned 18 species to this genus. But as it

was not delimited well, Vesicomya had long been

employed as a catchall taxon, to which many vesicomyids

from hydrothermal vent and cold seep ecosystems were

originally or subsequently assigned, sometimes rather

incongruently (Coan et al., 2000).

The name Vesicomya is often used between quotation

marks when the taxonomic status of a vesicomyine spe-

cies is not well resolved, e.g., as in “Vesicomya” filatovae

Krylova and Kameney, 2015 (Krylova et al. 2015). Con-

versely, some of its included species were assigned even to

different families or genera such as Kelliella or Callo-

cardia. Krylova et al. ( 2018) re-clarified the relationship

between Vesicomya and Kelliella, and assigned 15 species

to Vesicomya.

The family Verticordiidae Stoliczka, 1870 is another

common group of deep-water bivalves. Most members

are mobile infaunal carnivores that exclusively inhabit the

deep-sea (Coan and Valentich-Scott, 2012). At least 11

genera and more than 90 species have been reported for

eee eee eee

J. Jiang et al., 2019

Page 95

this family (Huber, 2010). Spinosipella Iredale, 1930 is

one of species-poor genus of verticordiids. Simone and

Cunha (2008) comprehensively revised this genus by

studying both conchological and anatomical characters.

Nowadays, five valid species are assigned to the genus: S.

acuticostata (Philippi, 1844), S. agnes Simone and Cunha,

2008, S. tinga Simone and Cunha, 2008, S. deshayesiana

(P. Fischer, 1862) (= Verticordia japonica A. Adams,

1862= Verticordia ericia Hedley, 1911), and S. coste-

minens (Poutiers, 1981).

Vesicomyids are well represented in the northern Pa-

cific (Krylova and Janssen, 2006), but the biodiversity in

Chinese waters is poorly known. Up to now, only two large

species of this family were described from the South China

Sea, Laubiericoncha nanshaensis (Xu and Shen, 1991) and

Archivesica marissinica (Chen, Okutani, Liang and Qiu,

2018). However, no Vesicomya species were found. Re-

garding Verticordiidae, a total of 6 species have been re-

ported from Chinese waters (Liu, 2008). Among them, two

species belong to genus Spinosipella i.e., S. deshayesiana

and S. costeminens (Liu, 2008; Xu and Zhang, 2008).

During the recent survey HYIV20150402_ to the

Qiongdongnan Basin on the northwestern slope of the

South China Sea by R/V Hal YANG Si Hao, anew vesicomyid

belonging to Vesicomya and a new verticordiid belong-

ing to Spinosipella were collected. Herein, we described

them as new.

MATERIALS AND METHODS

Specimens studied were collected by bottom trawling

with RV Hat YANG Si Hao from recently discovered Haima

Methane Seep about 1,400 m depth off southern Hainan

Island in the northern sector of the South China Sea, April

9°41 9015 (Liang et al., 2017; Feng et al., 2018). Together

with the specimens, this trawling also sampled 134 in-

dividuals of Bathymodiolus platifrons Hashimoto and

Okutani, 1994, and 9 of Archivesica marissinica (Chen,

Okutani, Liang and Qiu, 2018), which are typical vent and

seep specialized chemosymbiotic bivalves. Figure 1 shows a

map of the collecting site.

Shells were observed using a Zeiss Discovery V12 stereo

microscope. Photographs were taken using a Cannon

EOS6D camera or a Zeiss AxioCam 503 digital camera

coupled to the microscope. Measurements were made

with a Vernier caliper to the nearest 0.1 mm. All speci-

mens were collected dead and dried by Dr. Xuebao He, so

hindering examination of soft parts. Type specimens are

deposited in the Third Institute of Oceanography, Min-

istry of Natural Resources, Xiamen, China. The termi-

nology for morphological description and hinge teeth of

Vesicomyidae was used following Cox (1969), von Cosel

and Salas (2001), Krylova and Janssen (2006), Amano and

Kiel (2007). Abbreviations: SL, shell length; SH, shell

height; SW, shell width.

Figure 1. Map of the South China Sea showing the location of Haima Methane Seep, indicated by a rectangle (modified from Liang

et al., 2017).

Page 96

THE NAUTILUS, Vol. 133, Nos. 3-4

SYSTEMATICS

Superfamily Glossoidea Gray, 1847 (1840)

Family Vesicomyidae Dall and Simpson, 1901

Subfamily Vesicomyinae Dall and Simpson, 1901

Genus Vesicomya Dall, 1886

Callocardia (Vesicomya) Dall, 1886: 272.

Type Species: Callocardia atlantica Smith, 1885 (by

original designation).

Vesicomya rhombica new species

(Figures 2-7)

Description: Shell small, 7.10-8.20 mm long, fragile,

thin, rhombic in shape, inequilateral, equivalve. Anterior

dorsal margin depressed, concave; anterior end evenly

rounded; posterior dorsal margin sloping, slightly convex;

posterior end faintly angulate; ventral margin long and

rather convex. mbones protruding, ascending: ibrealeg

prominent, strongly prosogyrate, situated at the middle of

dorsal margin. Cue face white, porcellaneous, sculpture of

commarginal striae and low regular ribs. Periostracum

thin, glossy, translucent. Slight furrow running from umbo

to postero- -vental margin present on posterior area of

shell. Lunule triangular, sunken, distinct, demarcated by

incision. Rsentdheon indistinct. Ligament external, opis-

thodetic, strong, 1/3 length of posterior dorsal margin,

attached to nymphs on outer edge of posterior hinge plate.

Internal surface of valves white, smooth. Hinge plate

narrow. On right valve, anterior cardinal tooth 1 wedge-

like, located on ventral margin of hinge plate, its elongated

high posterior end areata! under FuNierstore ramus 3a of

posterior cardinal tooth, posterior ramus 3b thicker, ra-

diated posteriorly. On left valve, anterior cardinal tooth

long, bifid, V-shaped, with anterior 2a tooth long, along

ventral mar gin of hinge plate, fused with anterior ‘edge of

middle 2b tooth; posterior cardinal tooth 4b elongated,

parallel to postero-dorsal margin of the shell. Naleknatiare

muscle scars distinct, anterior teardrop-shaped, anterior

pedal retractor scar small, impressed, located dorsally to

and fused with anterior adductor scar; posterior adductor

scar reniform, larger than the anterior one; Pallial line

without pallial sinus.

Type Material: Holotype: TIO-BTS-MOL-1601

(Sta.QDN31), 1 specimen, SL $8.20 mm, SH 6.60 mm,

SW 4.40 mm. Paratype: TIO-BTS- MOL-1602(Sta.QDN Bil),

1 specimen, SL 7.10 mm, SH 5.60 mm, SW 3.80 mm. All

from type locality.

Type Locality: Haima Methane Seep, muddy bottom,

about 1,400 m depth off southern Hainan Island in the

northern sector of the South China Sea.

Distribution: Known only from type locality.

Etymology: The Latin specific epithet rhombica refers to

the rhombic shell shape in this species.

Remarks: The new species can be distinguished from

other congeners by its rhombic shell shape and a slight

furrow at the surface posterior area running from the

umbo to the postero-vental margin. Among the 15 spe-

cies within this genus assigned by Krylova et al. (2018),

seven of them are distributed in the Indo-West Pacific:

Vesicomya galatheae (Knudsen, 1970), V. pacifica (E.A.

Smith, 1885), V. tasmanensis (Knudsen, 1970), V. bruuni

Filatova, 1969, V. sundaensis (Knudsen, 1970), V. profundi

Filatova, 1971, and V. sergeevi Filatova, 1971 (Krylova

et al., 2015). In addition to differences in shell shape, the

first five species differ from the new species by the ar-

rangement of the ventral anterior cardinal tooth 1 and

anterior ramus 3a on the right valve, which do not overlap.

Instead, in the new species, the elongated high posterior

end of the cardinal tooth 1 is located under anterior ramus

3a of the posterior cardinal tooth on the right valve. The

hinge of the new species is very similar to those of

V. profundi and V. sergeevi, both of which are found in

the Kuril-Kamchatka Trench, northwest Pacific (Krylova

et al., 2015; Kamenev, 2019). But they differ by their shell

shape and sculpture. The shell of V. profundi is subcircular

in outline, higher, more inflated, and shorter in its anterior

and posterior ends, with the umbo protruding. Both the

new species and V. sergeevi have less inflated and longer

shells. But V. sergeevi is ovoid in shape, has less conspic-

uous commarginal ribs, and lacks the furrow on the pos-

terior area. Vesicomya indica (Knudsen, 1970), first found

in the central Indian Ocean, was also reported from

southern Shikoku, Japan (Tsuchida, 1994; Higo, Callomon

and Goto, 1999). But from the published figures (Tsuchida

1994: 78, fig. 3, pl. 3, figs. 1, 2), we can ascertain that it is

definitely not V. indica. Another species from Japan,

V. katsuae Kuroda, 1952 was assigned to Waisiuconcha

by Higo, Callomon, and Goto (1999: 2001). Krylova and

Janssen (2006) observed that it should be excluded from

the genus Waisiuconcha. Huber (2010) allocated the

species in the genus Isorropodon. Okutani (2017) still

included the species in Vesicomya, as originally de-

termined by Kuroda (1952). However, from the type

figure (Kuroda, 1952: 4, text- figs. 5-9; Higo, Callomon

and Goto, 2001: 174, fig. B1099), we believe that it

may belong to the genus Pliocardia, pending further

confirmation.

Superfamily Verticordioidea Stoliczka, 1870

Family Verticordiidae Stoliczka, 1870

Genus Spinosipella Iredale, 1930

Spinosipella Iredale, 1930: 388

Type Species: Verticordia ericia Hedley, 1911 (original

designation).

Spinosipella xui new species

(Figure 8-13)

Description: Shell small, 4.7 mm in length, inflated,

quadrangular or square-rhombus shaped, inequilateral,

weakly coiled, inequivalve, right valve slightly larger and

J. Jiang et al., 2019 Page 97

Figures 2-7. Vesicomya rhombica new species, holotype (TIO-BTS-MOL-1601). 2. Exterior of left valve. 3. Interior of left valve.

4. Exterior of right valve. 5. Interior of right valve. 6-7. Details of hinges. 6. Right valve. 7. Left valve.

Page 98 THE NAUTILUS, Vol. 133, Nos. 3-4

Figures 8-13. Spinosipella xui new species, holotype (TIO-BTS-MOL-1701). 8. Exterior of left valve. 9. Interior of left valve. 10.

Exterior of right valve. 11. Interior of right valve. 12-13. Details of hinges. 12. Right valve. 13. Left valve.

J. Jiang et al., 2019

Page 99

Figures 14-17. Semi-schematic line drawings of hinge plates. 14. Left valve of Vesicomya rhombica new species. 15. Right valve

of Vesicomya rhombica new species. 16. Left valve of Spinosipella xui new species. 17. Right valve of Spinosipella xui new

species.

overlapping left one, not gaping. Color chalky-white.

Antero-dorsal margin short, depressed-concave; postero-

dorsal margin sloping, slightly convex, about twice wider

than anterior one; anterior and posterior ends forming

abrupt right angles, with posterior one more protruded;

ventral margin zigzag crenulated, rather convex, with

antero-ventral margin almost vertical to postero-ventral,

tips of zigzag coinciding with tips of each surface rib, tips

corresponding to concavity of opposite valve; antero-dorsal

margin almost parallel to postero-ventral, and postero-

dorsal margin parallel to antero-ventral, forming lozenge-

shaped shell outline. Umbones coiled, convex, projected,

divergent, prosogyrate, situated in middle of dorsal margin.

Outer surface with minute, opaque granulations forming

irregular mosaics. Sculptured by strong, arched, widely

spaced, 13 radial ribs triangular in cross-section, forming

broad undulations on surfaces of valves and producing

deeply plicate margins;

largest rib on middle forming prominent keel, dividing

valve into anterior and posterior regions each with 6 ribs;

ribs gradually becoming smaller from middle to shell

ends. Lunule cordate, sunken, distinct, demarcated by

incision. Escutcheon obscure, long, lanceolate. Internal

surface of valves glossy, nacreous, white, iridescent, with

undulations corresponding to ribs on external surface.

Hinge plate narrow, under lunule. On right valve, hinge

with large, stubby, tall, conical cardinal tooth, and long

lamellar posterior lateral tooth parallel to postero-dorsal

shell margin. On left valve, anterior cardinal tooth stout,

long, extending along antero-dorsal margin, almost con-

nected to smaller anterior lateral tooth under lunule;

posterior cardinal tooth low, short and wedge-shaped;

shallow socket between cardinal teeth, restricted to dor-

sal surface, and corresponding to the cardinal tooth of right

valve. Anterior adductor muscle scars distinct, reniform,

posterior adductor muscle scars elongate-ovate.

Ligament opisthodetic, weak, almost imperceptible,

nearly fused to periostracum along dorsal hinge margin;

resilium brown, posterior to cardinal teeth. Pallial line

very poorly defined.

Type Material: Holotype: TIO-BTS-MOL-1701

(Sta.QDN31) SL 4.7 mm, SH 4.3 mm, SW 3.8 mm.

Type Locality: Haima Methane Seep, muddy bottom,

about 1,400 m depth off southern Hainan Island in the

northern sector of the South China Sea.

Distribution: Known from the type locality.

Etymology: This species is dedicated to Prof. Fengshan

Xu for his great contributions to Chinese malacology.

Remarks: The new species is allocated to the genus

Spinosipella Iredale, 1930, due to its outline, a single

strong cardinal tooth in the right valve, and a shallow

socket in the left valve corresponding to the cardinal tooth

of the right valve (Poutiers and Bernard, 1995). It is also

noteworthy that the new species is different from other

species of the genus in having a long thin lamellar pos-

terior lateral tooth on the right valve. A long posterior

lateral tooth on the right valve is also one of the characters

typical of the genus Trigonulina dOrbigny, 1853 (Poutiers

and Berard, 1995: Coan and Valentich-Scott, 2012).

However, the linule of the new species is sunken, distinct,

and well-impressed, but not so deeply depressed as the

two species of Trigonulina, i.e., the eastern Pacific

Trigonulina novemcostata (A. Adams and Reeve, 1850)

[= Trigonulina pacifica Jung, 1996= Trigonulina han-

cocki (Bernard, 1969)| and the western Atlantic Trig-

onulina ornata d Orbigny, 1853. The lunules of these two

species are deeply incised into the shell and overhung by

umbones, and the hinge plates thickened. At the moment,

only two species of Spinosipella with a tropical West

Page 100

Pacific distribution are known (Simone and Cunha, 2008;

Xu and Zhang, 2008). The number of radial ribs in the new

species is much less than these two similar species, which

have about 18-19 and 16-17 ribs, respectively. The new

species has only 13 radial ribs, which is comparable to the

Mediterranean S. acuticostata (Simone and Cunha, 2008).

But S. acuticostata is higher than the new species, with

stronger ribs and more prominent spines on the surface.

ACKNOWLEDGMENTS

The authors are grateful to Gene Coan, Steffen Kiel, and

Elena Krylova for comments and suggestions on

the manuscript, to Konstantin A. Lutaenko for providing

some important literatures, to Xuebao He (Third Institute

of Oceanography, Ministry of Natural Resources), who

collected and loaned the material for this study, and to the

scientific staff of the expedition and the ship crews of R/V

HAI YANG SI HAO. This research was supported by the Basic

Science Research Fund of Third Institute of Oceanogra-

phy, MNR (No. TIO2016043), the National Special Project

on Gas Hydrate of China (No. DD20190218), the Strategic

Priority Research Program of the Chinese Academy ee

Sciences (No. XDA22050203), the Special Funds for

Young Scholars of Taxonomy of the Chinese Academy of

Sciences (No. ZSBR-009), and the Ocean Public Welfare

Scientific Research Project, State Oceanic Administration

of the People’s Republic of China (No. 201505004-4).

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