Olivera Et Al 2014 Annual Review

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Biodiversity of Cone Snails and Other Venomous Marine Gastropods:

Evolutionary Success Through Neuropharmacology
Article in Annual Review of Animal Biosciences · February 2014

DOI: 10.1146/annurev-animal-022513-114124 · Source: PubMed
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Neuropharmacology
Baldomero M. Olivera,1 Patrice Showers Corneli,1
Maren Watkins,1 and Alexander Fedosov2
1
Biology Department, University of Utah, Salt Lake City, Utah 84112;
email: olivera@biology.utah.edu, corneli@biology.utah.edu, maren.watkins@hsc.utah.edu
2
A.N. Severtzov Institute of Ecology and Evolution, Russian Academy of Science,
Moscow, 119071 Russia; email: fedosovalexander@gmail.com
Annu. Rev. Anim. Biosci. 2014. 2:487–513 Keywords

First published online as a Review in Advance on diversity, gene families, Conoidea, venom peptide
November 14, 2013
The Annual Review of Animal Biosciences is online Abstract

at animal.annualreviews.org
Venomous marine snails (superfamily Conoidea) are a remarkably bio-
This article’s doi: diverse marine invertebrate lineage (featuring more than 10,000 species).
10.1146/annurev-animal-022513-114124
Conoideans use complex venoms (up to 100 different components for
Copyright © 2014 by Annual Reviews. each species) to capture prey and for other biotic interactions. Molecular
All rights reserved
phylogeny and venom peptide characterization provide an unusual mul-
tidisciplinary view of conoidean biodiversity at several taxonomic levels.
Venom peptides diverge between species at an unprecedented rate
through hypermutation within gene families. Clade divergence within
a genus occurs without recruiting new gene families when a saltatory
event, such as colonization of new prey types (e.g., fish), leads to
a new radiation. Divergence between genera in the same family involves
substantial divergence in gene families. In the superfamily Conoidea, the
family groups recruited distinct sets of different venom gene superfami-
lies. The associated morphological, behavioral, and prey-preference
changes that accompany these molecular changes are unknown for most
conoidean lineages, except for one genus, Conus, for which many
associated phenotypic changes have been documented.
487
INTRODUCTION
More than 10,000 species of venomous marine snails comprise three major families within the su-
perfamily Conoidea (formerly Toxoglossa, or poison tongue) including the best-known and most
intensively studied, the Conidae (the cone snails, genus Conus). Along with the two other major groups
of venomous gastropods, the auger snails (family Terebridae) and the turrids (previously assigned to
the traditional family Turridae, sensu lato), these conoideans (Figure 1) include highly specialized
predators with diverse feeding guilds (1, 2). The striking shell patterns have long attracted a large cadre
of collectors; in the seventeenth century, cone shells were among the most valuable of natural history
objects (see sidebar, Cone Shells: Treasures and Art Objects). Conoideans comprise a monophyletic
grouping (3, 4) assigned to the infraorder Neogastropoda (5), a large group of mainly carnivorous
marine snails that underwent a remarkable radiation in the Cretaceous and early Cenozoic (6, 7).
In every respect, the taxonomy, evolutionary history, ecology, and biochemistry of the family
Conidae have been characterized far more extensively than those of the auger snails and turrids (1,
2, 8, 9). Furthermore, because of the neuropharmacological interest in their venom components

(10), a broader spectrum of species with unique gene products likely are defined in Conus than
in any other genus of animals, which provides a species-specific window into both their evolution
and the physiological differences between species (see sidebar, Present and Future Biomedical
Applications of Marine Snail Venoms). Here we present an overview of the scientific literature on
this very successful, though largely still unknown, group of animals. As is the case for all shelled
molluscs, the traditional taxonomy and phylogeny of the group has been, until recently, largely
based on shell morphological characters. We outline the rapidly expanding biochemical, mo-
lecular, and morphological data and discuss the pertinent literature on the biology of enven-
omation of this extremely speciose and diverse group of venomous marine snails.
SYSTEMATICS AND PHYLOGENY OF CONOIDEA

Systematics and Phylogeny of the Three Conoidean Families
Among the three traditional groups in the Conoidea, the cone snails and augers are easily rec-
ognizable from their characteristic shell morphology (Figure 1) and have consistently been
CONE SHELLS: TREASURES AND ART OBJECTS
Cone shells have long been objects of admiration. They were found with carnelian beads apparently strung into
necklaces at Uruk, the first human urban settlement in Mesopotamia, dated to more than 5,000 years ago. In the
eighteenth century, the Dutch brought cone shells back from Indonesia, which triggered a shell-collecting craze.
Rare cone shell species were regularly sold at auction; one rare species, Conus cedonulli (“the matchless cone”),
brought four times the price of a seventeenth-century painting by Vermeer (Woman in Blue Reading a Letter) sold at
the same auction in 1796 (Conchylomania, http://penelope.uchicago.edu).
The Dutch still-life painters of the period included cone shells in their compositions with flowers and fruits. In
1650, Rembrandt, a contemporary of Vermeer, did etchings of the marble cone, Conus marmoreus, which he
continued revising throughout his life. The image is etched and inked on a metal plate and printed on paper in
a mirror image of the plate. Although he remembered to reverse his signature, he did not reverse the chirality of the
shell. Hence, the printed image on paper is a sinistral shell, whereas all natural shells of C. marmoreus are dextral
(Curatorial Notes from the British Museum, http://www.britishmuseum.org/explore/highlights/highlight_objects/
pd/r/rembrandt,_the_shell.aspx)
488 Olivera et al.

assigned to two well-established family groups, Conidae and Terebridae, respectively. Although
there have been sporadic proposals to split these families into smaller units (e.g., Reference 11),
most of the taxonomic literature continues to assign all approximately 700 species of cone snails to
the Conidae and all 350–400 species of auger snails to the Terebridae (12–16). Traditionally, Lumun-lumun:
Filipino method in
conoidean species that were not cone or auger snails were assigned to the Turridae, which therefore
which tiny marine
was a group largely defined by exclusion (3, 17). Comprising the greatest conoidean biodiversity animals colonize
(>10,000 species), turrids are well known to malacologists for their problematic and unstable bundled fine-mesh nets
taxonomy. The leading turrid taxonomist of the twentieth century, A.W.B. Powell (18), sum- kept in deep water,
marized turrid taxonomy in his major treatise; with a total of more than 270 genera assigned to retrieved, and then sold
to microscope
Turridae, the group by far exceeded both the cone snails and the augers in complexity and
hobbyists
diversity.
The first analysis to provide evidence that the classical Turridae is not a monophyletic group
was based on foregut anatomy (19). This comprehensive anatomical study demonstrated that
some turrid subfamily groups had a closer affinity to cone snails than to Turris, the nominate genus
of the Turridae. Taylor & Morris (7) proposed to move some subfamilies and genera previously
assigned to Turridae to Conidae. An extensive analysis of radular morphology also led to a revised
phylogeny for turrids (5, 11). Thus, the family Turridae was restricted to a smaller set of genera.
Tucker & Tenorio (11) proposed eight new families, most equivalent to former subfamilies of
Turridae, including Borsonidae, Raphitomidae, Clathurellidae, Drillidae, Clavatulidae, and
Crassispiridae. These workers also suggested splitting Conoidea into two superfamilies, Conoidea
(Fleming 1822) and Turroidea (Swainson 1840).
The massive amount of new conoidean material collected in recent years has revealed that the
number of conoidean species has been grossly underestimated, primarily because a considerable
fraction of turrid diversity comprises micromolluscs and because the superfamily has undergone
an amazing radiation in deep-water habitats. Naming of all of the recently collected distinct
morphospecies from deep-water scientific expeditions organized by Bouchet et al. (20–22) and by
a new method for collecting micromolluscs developed in the Philippines, called lumun-lumun (23),
has been limited by the availability of taxonomists with the appropriate expertise. The expanding
molecular data sets now encompass an increasing diversity of forms sufficient for delineating the
major turrid lineages. The recent analysis by Puillandre and coworkers (3, 17), shown in Figure 2,
creates 15 different families in the superfamily Conoidea.
PRESENT AND FUTURE BIOMEDICAL APPLICATIONS OF MARINE SNAIL VENOMS
An unusual facet of the venoms of cone snails and other members of the superfamily Conoidea is their validated
biomedical potential. One venom component, from the fish-hunting Conus magus, the magician’s cone, is an
approved drug for severe, intractable pain (commercial name, PrialtÒ; generic name, Ziconotide) (88). Although
the commercial drug is chemically synthesized, it is identical to the peptide made in the snail’s venom duct. The
biotechnology company that developed Prialt spent two years modifying chemical groups in an effort to optimize
pharmacological properties; in the end, the compound they decided to move forward with into production is
identical to that made by the snails—not a single chemical group was changed. Five other peptide toxins from
cone snail venoms have reached human clinical trials; an even larger number are at preclinical stages of
development for diverse potential therapeutic applications. Only a miniscule fraction of the total biodiversity of
venomous marine snails has been examined, and the potential for future biomedical applications is thus very
much greater.
www.annualreviews.org Venomous Marine Snails 489

a b
d e
c
Conus imperialis Conus marmoreus g

f
h i
k n
l m
Terebra
triseriata Turris Turris
Terebra grandis
subulata babylonia
Figure 1
(Left panel) The three classical families in Conoidea. Shown are two cone snails, Conus marmoreus, the marble cone, which is the type
species in the genus Conus, and Conus imperialis, the imperial cone. The shells of two auger snails are illustrated, Terebra subalata, the type
species of the genus Terebra, and Terebra triseriata, a deep-water species from the Indo-Pacific. The shells of two turrids are also shown:
Turris babylonia, the type species of the genus Turris, and Turris grandis, the species that grows to the greatest length in this genus. (Right
panel) Shells of turrid species formerly in Turridae now assigned to other families. (a,b) Drillidae: (a) Clavus rugizonatus, (b) Plagiostropha
sp. (c) Marshallena philippinarum (Horaiclavidae). (d) Cochlespira pulchella (Cochlespiridae). (e) Funa sp. (Pseudomelatomidae).
(f) Perrona lineata (Clavatulidae). (g,h) Borsoniidae: (g) Tomopleura sp., (h) Zemacies excelsa. (i) Glyphostoma sp. (Clathurellidae).
(j,k) Mangeliidae: (j) Eucithara sp., (k) Hemilienardia thyridota. (l–n) Raphitomidae: (l) Gymnobela sp., (m) Veprecula polyacantha,
(n) Pseudodaphnella granosa.
Cone Snails: Recent Advances in Taxonomy and Phylogeny

The cone snails (family Conidae) are by far the best-known, most intensively investigated con-
oidean lineage. Despite continuous attempts to divide the approximately 700 species of cone snails
into smaller generic groups (11), until recently, the general approach was to assign all living cone
snail species to a single genus (Conus), regarded as the only extant genus in the family Conidae,
490 Olivera et al.

along with several fossil genera (however, see References 11, 17). The expanding molecular
database for cone snails (24–33) revealed that a significant fraction of cone snail species are only
distantly related to the majority of Conus: Three groups are sufficiently divergent to justify
separation from the genus. In this review, these three distinct lineages will be regarded as separate Radula: chitinous,
ribbon-shaped rasping
genera in the family Conidae: Conasprella (Thiele 1929), Profundiconus (Kuroda 1956), and
structure covered with
Californiconus (11) (see Figure 3). rows of teeth
The genus Californiconus has a single known species, Californiconus californicus, found off the (denticles) used for
coast of California to the Baja California peninsula of Mexico. Profundiconus appears to be the feeding and adapted to
most basal living clade in the family Conidae and includes little-known deep-water cone snail deliver venom in
conoideans
species found in the Indo-Pacific. The third of the living genera is Conasprella, which appears to
have a worldwide distribution.
A more detailed phylogenetic tree for the genus Conus, which includes more than 80% of all
living species in the family Conidae, is presented in Figure 4, along with the primary prey for species
in each clade. Mapping of the dietary guilds on Conus phylogenetic trees (e.g., References 28, 30)
suggests that the ancestral cone snails were worm-hunting species. And although the majority of
conids prey on vermiform invertebrates (primarily polychaetes), distinct clades of Conus de-
veloped fish-hunting behaviors. These are not monophyletic, suggesting that fish hunting
appeared more than once in the evolutionary history of Conus. Unlike fish hunters, snail-hunting
Conus species form a monophyletic group; therefore, the transition to molluscivory was likely
a single evolutionary event.
BIOLOGY OF ENVENOMATION
Anatomy and Venom Delivery Strategies
Conoideans use venom as the primary weapon for capturing prey. The venom is produced in
a specialized tubular esophageal gland, the anatomical signature of the superfamily Conoidea.
Appearing early in conoidean evolution, the unique mechanism of envenomation seems to be
a prerequisite of their evolutionary diversity.
The distal part of the venom gland forms a muscular bulb, which contracts to extrude the
venom from the venom gland. Because of the complex structure of the apparatus, the proximal
tube is often referred to as a venom duct, even though all venom components are synthesized and
secreted there. The venom gland opens into the pharynx (i.e., buccal mass), from which venom
flows into the large, extensible proboscis. The highly modified radula plays a key role in enven-
omation. Typically represented by a flexible band (the radular membrane) with segmentally
chitinous plates (radular teeth), the gastropod radula functions as a rasping organ. In the course of
conoidean evolution, however, the radula in some species has evolved to form a variety of re-
markably effective venom delivery tools.
In cone snails and most closely related families, the radular membrane is reduced, and each
radular tooth is strongly enlarged and shaped as a hollow hypodermic needle (see Figure 5).
Typically, one radular tooth is detached from the membrane and moved to the tip of the extended
proboscis as the snail strikes it prey. During envenomation, a single radular tooth travels to the
proboscis tip, where it is held by specialized muscular sphincters (34, 35). As the snail extends its
proboscis toward the prey, the radular tooth is used to pierce the prey’s tegument and inject venom.
Often, radular teeth of cone snails, especially those that hunt fish, bear barbs and blades at their tips
and strongly resemble a harpoon (Figure 5e). As the harpoon-shaped end pierces the scales and
skin, the barbs tether the fish (see Figure 6). This process has been described for several Conus
species (2, 36, 37), recently in mechanistic detail for Conus catus (38, 39). In effect, the radular

tooth serves as both a disposable hypodermic needle for injecting venom and a harpoon to tether
fish. Various facets of the radular tooth morphology have been used as important characters for
the cladistic analysis of cone snails (11, 40). Although all cone snails have hollow, hypodermic
radula (34), considerable variation exists in the detailed morphology and in how such teeth are
used. In fish-hunting cone snails, only one hypodermic tooth is used in each prey-capture event. In
contrast, many mollusc-hunting cone snails require multiple injections of venom before they can
begin to devour their prey. For example, Conus ammiralis has been observed to harpoon its snail
prey more than half a dozen times.
In several other major groups, including Drillidae, Turridae, Clavatulidae, and Psuedomela-
tomidae, the radula retains its integrity, and radular teeth are solid. Kantor & Puillandre (41)
suggested that a solid radular tooth is lodged at the tip of the proboscis and is used to stab the prey,
presumably to apply venom in the stab wound.
Correlation of morphologic data with molecular phylogeny has allowed researchers to trace
radular evolution in major clades of the superfamily Conoidea (42). In particular, the hollow
radular teeth appear to have evolved independently in the families Conidae and Terebridae; in the
latter, evolution of hypodermic-type teeth may have occurred more than once (12). Altogether, the
family Terebridae, a well-recognized monophyletic group, is characterized by a surprising di-
versity of venom-delivery strategies, reflected in diverse radular morphologies. Some terebrids
have solid teeth; in other lineages, the functions of unusual teeth morphologies are not well
understood (Figure 5). Finally, some terebrid lineages lack the specialized foregut complex
(i.e., radula, proboscis, and venom apparatus) entirely. More remarkably, the loss of these foregut
structures has occurred multiple times in Terebridae (43). Terebridae is not the only family that has
lost venom-associated foregut structures; this was also documented in members of at least three
other families, including Horaiclavidae (44) and Borsoniidae (45), and several times independently
in Raphitomidae (46, 47).
Venom Composition
The venoms of conoidean gastropods are invariably complex; typically each species can express
a repertoire of 50–200 peptide toxins in its venom. In cone snail venoms, most peptides are small (12–
35 AA), with multiple disulfide crosslinks (48) and a suite of distinctive posttranslational mod-
ifications. The complement of peptide toxins in the cone snails may be somewhat smaller in size than
in some of the other lineages, where small proteins, which are invariably also multiply disulfide
bonded, comprise a larger fraction of the spectrum of venom components. A striking feature of these
venoms is that each species has its own distinct complement of peptide toxins (49), which implies very
rapid divergence, even between closely related species. Each peptide toxin has a specific, physio-
logically relevant protein target, typically a signaling molecule on the neuronal surface.
Behavior observed in aquaria (see Supplemental Videos 1–3 at the Supplemental Material link
Supplemental Material in the online version of the article or at http://www.annualreviews.org/) suggests that venom is not
exclusively used for prey capture but may also be used for defensive purposes against potential
predators and for competitive interactions (see Supplemental Video 4). In effect, venom reflects the
Figure 2
→
Maximum likelihood tree for Conoidea inferred from COI, 12S, and 16S sequences [tree file from Puillandre et al. (3)]. The companion
taxonomic proposal (24) divides turrids into 13 family groups, creating 15 different families in the superfamily Conoidea. Asterisks
indicate clades with both a Bayes posterior probability and a bootstrap value greater than 95%. Circles show clades with at least one
support value greater than 95%. Colors and labels show family status according to the classification of Bouchet et al. (17).
492 Olivera et al.

Xenophora
Laevistrombus
Belomitra
Turrillatirus
Vexillum
Harpa Profundiconus, Conidae
Profundiconus
Californiconus Californiconus, Conidae
Conasprella
Conus Conasprella, Conidae
Taranteconus Conus, Conidae
Clathurella
Nannodiella Clathurellidae
Etrema
Benthofascis Corneorbidae
Microdrillia
Borsoniidae genus 1
Typhlomangelia Borsonidae
Genota
Borsonia
Tomopleura
Zemacies
Bathytoma Mitromorphidae
Lovellona
Mitromorpha
Benthomangelia
Toxicochlespira
Oenopota
Anticlinura
Mangelliidae genus 1
Eucythara
Heterocithara Mangellidae
Mangelliidae genus 2
Hemilienardia
Raphitoma
Veprecula
Rimosodaphnella
Glyphostomoides
Tritonoturris
Eucyclotoma
Kermia
Pleurotomella Rhaphitomidae
Taranis
Daphnella
Teretiopsis
Thachteria
Gymnobela
Cochlespira
Sibogasyrinx
Cruziturricula
Fusiturricula Cochlespiridae
Drillia
Clathrodrillia
Agladrillia
Imaclava
Calliclava
Conopleura Drillidae
Splendrillia
Iredalea
Clavus
Cerodrillia
Leucosyrinx
Comitas
Knefastia
Pseudomelatoma
Tiariturris
Otitoma
Carinodrillia
Pyrgospira Pseudomelatomidea
Pilsbryspira
Zonulispira (Crassispiridae)
Hindsiclava
Cheungbeia
Funa
Inquisitor
Ptychobela
Crassispira
Gemmuloborsonia
Turricula
Pusionella
Perrona Clavatulidae
Clavatula
Paradrillia
Carinapex
Ceritoturris
Anguloclavus Horaiclavidae
Horaiclavidae genus 1
Anacithara
Horaiclavus
Lucerapex
Polystira
Ptychosyrinx
Turridrupa
Lophiotoma
Gemmula Turridae
Turris
Xenuroturris
Iotyrris
Euterebra
Hastula
Oxymeris
Cinguloterebra
Clathroterebra
Hastulopsis
Terenolla Terebridae
Myurella
Terebra
2.0 substitutions Strioterebrum
per site

Conus
pu
molu
atus
radiatus
rpu
us
croc
ra
ccen
at
erm s
ticu
en
sce
ine
sp eris
ar
on
act
ns
sis
sa us
lis 100
car
music 100 100
us 100
us
sulcatu 100 striat
s 100 100
100 aurisiacus
99
geographus 100
con
sor
s
ma
100 100/88
99
g
s
ebraeu s
us
100
a l deu Californiconus
ch lu
m
chia alis
xil
100/100
us
ngi
ve
ne
californ
i
per
icus
ita
im
cap
100/100
100/96
100/–
arcuatus
chii
Profundiconus
100
ion
ma
100/90 100/96 m e
tera 100/96 pa emia
go e
du
100/99 s
jas
pe
ichnose
diu
marnatus
to
s
orbignyi
is
ho
urions
u
ga
ana
ine
ni
vim
long
Conasprella
0.3 substitutions
per site
Figure 3
Bayesian tree inferred from cytochrome oxidase I and intron 9 of the g carboxylase gene for representative
members of the family Conidae showing very large evolutionary change separating the four distinctive groups
classically belonging to the genus Conus (for methods see Kraus et al. (28)]. Branches joining the four genera are
labeled with Bayesian posterior probabilities (left) and approximate likelihood ratio test (aLRT) values from the
maximum likelihood analysis (right). Based on the large evolutionary distance, we refer to the other clades as
additional conid genera: Conasprella, Californiconus, and Profundiconus. Californiconus californicus is the
only extant species known in this genus. Profundiconus species are found in extremely deep water in the Indo-
Pacific, whereas Conasprella is a worldwide group, generally found offshore in moderately deep water in the
Indo-Pacific, though this genus has successfully colonized shallow-water habitats in the Panamic and Caribbean
regions.
chemical interactions of a particular conoidean species with the prey, predators, and competitors it
encounters in its ecological niche.
Molecular Physiology of Envenomation

Analyses of the predator/prey interaction of cone snails that belong to two fish-hunting lineages,
Pionoconus and Chelyconus, provide the most detailed picture of venom physiology (see the
following section). Overall, venom physiology is characterized by multiple toxins that selectively
target a signaling protein present on the plasma membrane and act in concert to dramatically
494 Olivera et al.

subvert the normal physiology of the prey. Teichert et al. (50) referred to this strategy as
constellation pharmacology. Briefly, a group of diverse toxins synergistically attack a set of
molecular targets, which are functionally linked within a specific physiological circuit (48, 51).
These synergistic toxin groups are referred to as cabals (1, 52), in reference to secret societies out to Constellation:
a functionally coupled
overthrow existing authority. The set of molecular targets of a cabal are known as the com-
set of signaling proteins
plementary constellation of ion channels/receptors/transporters (53). Thus, for every cabal in the (receptors and ion
venom, there is a complementary constellation of targets in the envenomated prey. channels) in a cell that
Not surprisingly, one widely targeted constellation is made up of the ion channels and receptors are targeted by the
that control neuromuscular transmission, a critical circuit for prey capture. The relevant cabal of cabal
toxins acts on well-known signaling components in the neuromuscular circuitry (e.g., voltage- Cabal: a group of
gated Ca2þ channels at the presynaptic terminus, the nicotinic acetylcholine receptor at the venom peptides acting
synergistically to affect
postsynaptic terminus, and the voltage-gated Naþ channels on the muscle membrane that bring the
a particular
action potential from the neuromuscular synapse to the contractile apparatus). Thus, the evolution physiological response
of conoidean gastropods has produced what is in essence a highly sophisticated equivalent to in the envenomated
combination drug therapy. animal
DIVERSITY OF THE FISH-HUNTING CONE SNAIL CLADES
The Pionoconus Clade of Fish-Hunting Conus: Similarity and Divergence of Species

Alan Kohn’s (54) landmark studies on the biology of cone snails in the mid-twentieth century first
defined the diverse prey of cone snails; his results can be tightly correlated to the molecular phy-
logenetic data (25) elucidated decades later (Figure 4). At present, the significance of the phylogenetic
data in a wider biological context can best be illustrated by focusing on the fish-hunting clades in the
genus Conus; these are generally given subgeneric rank. Four of these clades, Pionoconus, Gas-
tridium, Textilia, and Phasmaconus, are found in the Indo-Pacific. One clade, Chelyconus, is found
exclusively in the New World. Kohn (36) first discovered and recorded fish hunting in conoideans.
He showed how the striated cone, Conus striatus, captured its fish prey.
At least 20 different Conus species belong in the subgenus Pionoconus (Figure 7), and an
additional 5–10 species are likely to be members of the clade. The biogeography varies greatly
between different Pionoconus (55); some species are distributed across the entire Indo-Pacific, but
others are highly restricted geographically. C. striatus, the largest species in the Pionoconus clade,
is found from East Africa to Hawaii. In contrast, Conus gauguini is endemic to the Marquesas
Islands and Conus barthelemyi to Mauritius/Reunion in the Indian Ocean.
In regard to shell morphology, Pionoconus species display a great diversity with striking
variation in coloration and shell pattern and considerable size divergence from species to species
(see Reference 55). Whereas C. striatus grows to more than 125 mm in length, Conus striolatus can
be relatively small (20–25 mm). The habitat of some Pionoconus, such as C. catus and Conus
stercusmuscarum, is in the intertidal zone. C. stercusmuscarum forages at night in isolated tide
pools at low tide. Other species (e.g., Conus circumcisus) are found only well offshore, in relatively
deep water (>30 m). Most species of Pionoconus have a free-swimming larval (veliger) stage of
a few weeks, after which they settle down. Conus magus and others have a much shorter pelagic
stage. Upon settling, juvenile Pionoconus individuals first consume worms then switch over to fish
hunting some weeks later (56).
All species of Pionoconus hunt fish, initially approaching their prey with a highly extended
proboscis. When stricken, the fish is tethered because the radular tooth used for injecting the
venom is barbed like a hook and line (see Supplemental Video 1). However, Pionoconus venoms Supplemental Material
contain two different sets of toxins that target different neural circuitry in the prey (1). One group,

100/98/100 consors
achatinus
100/95/100 magus P Pionoconus
100/93/100
100/100/100 circumcisus
aurisiacus
100/–/– gauguini
striatus
100/99/– dusaveli P Textilia
cervus
–/–/100 100/100/100 nux Vn Harmoniconus
musicus
100/98/84 obscurus
geographus P Gastridium
tulipa
laterculatus
100/100/99 moluccensis
–/84/85 P Phasmoconus
spectrum
100/91/– parius
100/90/– radiatus
–/100/91
caracteristicus Ve, n, c Puncticulis
–/–/100 arenatus
pulicarius
100/100/98 ermineus P Chelyconus
purpurascens
bandanus M Conus
100/93
marmoreus
aulicus
90/95/98
100/98/– episcopatus M Darioconus
98/93/98 crocatus
omaria
100/85/100 furvus
gloriamaris
ammiralis M Cylindrus
100/100/90 textile
100/100/–
dalli
ebraeus Ve, n Virroconus
100/100/–
chaldeus
quercinus Ve, t Lividiconus
lividus
100/92/– moreleti Vt Virgiconus
terebra
virgo
100/100/91 100/100/100 glans Ve Leporiconus
–/98/– nucleus
–/92/– tribblei Vn Kioconus
100/92/–
viola
regius
100/98/100 archon
chiangi Va Stephanoconus
100/–/100
–/97/– imperialis
100/100/99
zonatus
planorbis Vp Vituliconus
striatellus
capitaneus Ve Rhizoconus
vexillum
0.2 substitutions
per site
Figure 4
Maximum likelihood tree for the genera Conus inferred from partial cytochrome oxidase I and 12SrRNA sequences [for methods see
Kraus et al. (28)]. Branches are labeled with Bayesian support values (left); with aLRTs (center); and, where available, with support values
from analysis of intron 9 sequences (28). Clades are colored and labeled with subgeneric names and with diet. The rooting of the tree is based on
our previous work (28–30). Vermivory appears to be the ancestral feeding behavior for this genus, whereas the descendant piscivorous
clades comprise a polyphyletic group of five subgenera. In contrast, the molluscivory appears to have evolved just once. Abbreviations: a,
amphinomids; c, capitellids; e, eunicids; P, piscivorous; p, polychaetes; M, molluscivorous; n, nereids; t, terrebellids; V, vermivorous.
known as the lightning strike cabal, causes a very rapid immobilization of prey. These toxins act
very much like a Taser: It is as if the fish has been electrocuted, resulting in an immediate tetanic
paralysis after the venom is injected. The toxins of the second group act more slowly but more
irreversibly, causing a complete paralysis of skeletal muscles. This is known as the motor cabal; the
net effect is that the fish is hit with two sets of powerful pharmacological mixtures with two
different physiological endpoints, either of which is usually sufficient to immobilize the fish. This
strategy is shared by all directly observed Pionoconus species.
The individual toxins contained in venoms of a significant number of Pionoconus species have
been identified and are responsible for various effects on fish. Some species-specific peptide toxins,
similar to each other in structure, are clearly encoded by sets of homologous genes. However,
496 Olivera et al.

a b
Conotoxin: Conus
venom peptides,
typically containing
multiple disulfide
bonds, acting
synergistically to
modulate the activity of
ion channels,
0.1 mm 0.1 mm receptors, or
transporters
c 0.1 mm d
0.1 mm
1 mm
Figure 5
Example of radula of Conus and other conoideans. (a,b) Nonhypodermic radula. (c–e) Single hypodermic-type
radular teeth. (a) Clavus exasperatus (Drillidae); (b) Turris babylonia (Turridae); (c) Hastula hectica
(Terebridae); (d) Lienardia gilberti (Clathurellidae); (e) Conus bullatus (Conidae).
a significant divergence in amino acid sequences is observed, even for two peptides from closely
related species acting on the same molecular target. Examples of two groups of peptides used for
prey capture are shown in Table 1, one group (the a3/5-conotoxins) belonging to the motor cabal
and the other group (the d-conotoxins) to the lightning strike cabal.
Two Fish-Hunting Clades, Chelyconus and Pionoconus: Convergent Evolution

The physiological effects of venom on fish prey for two of the groups of piscivorous Conus, Pio-
noconus and Chelyconus, are mechanistically convergent. However, based on any of the standard
genetic markers for phylogeny, species in the Chelyconus clade are evolutionarily well separated
from the Indo-Pacific fish-hunting species (see Figure 4). The phylogenetic tree obtained by using
a very short intron 9 sequence from the g-glutamyl carboxylase gene has been diagnostic in sep-
arating Chelyconus from the other fish-hunting species, which in turn can be separated from one
another by this intron (28). Some classes of toxins used for the capture of fish prey are also strongly
evolutionarily divergent between the two clades. This appears to be a remarkable example of
convergent evolution: Fish hunting seems to have evolved from different presumptive worm-hunting
ancestors independently, ultimately evolving convergent physiological strategies but, in some cases,
recruiting different gene superfamilies to target the same ion channel in fish.
This convergent physiology is juxtaposed with a stark contrast in biodiversity levels of fish-
hunting species in the Indo-Pacific versus the New World. In the Indo-Pacific, the radiation of
the Pionoconus clade has resulted in over 20 living species (shown in Figure 7). In contrast, the

a
b e
c f
Figure 6
Four Conus species with their prey. (a–c) A series of three panels showing the attack of the fish-hunting
species Conus consors. (a) The snail has its proboscis extended and is about to strike the fish. (b) The fish
immediately after being envenomated; the snail has retracted its proboscis and is about to engulf the fish with
its rostrum (false mouth). Note the very stiff dorsal fin of the envenomated prey. (c) The fish is almost
completely engulfed by the C. consors. (d) Conus imperialis, a species in the Rhizoconus clade, extending its
bright red proboscis toward an amphinomid polychaete (fireworm). (e) The molluscivorous species Conus
ammiralis feeding on an envenomated prey snail, a columbellid. (f) Conus eburneus, extending its pale white
proboscis, about to attack the polychaete worm, Nereis. Note the difference in the size of the proboscis in
the two worm-hunting species.
Chelyconus clade comprises only two known species: Conus purpurascens, the only species in the
entire Panamic region, and Conus ermineus, the only Chelyconus in the entire Atlantic Ocean. To
discover why the evolution of fish hunting in the Indo-Pacific led to a rich harvest of speciation
events, and why this was not the case in the Chelyconus radiation, we must address key factors that
promote the generation of new species in a lineage within a given evolutionary period. In the case of
Pionoconus speciation, Williams & Duda (57) suggest that tectonic activity in the Indo-West
Pacific during the late Oligocene/early Miocene created a rich mosaic of distinct habitats that
promoted a rapid diversification of the shallow-water biota in the marine environment. This is
consistent with the contrast between Pionoconus, a clade found mostly in the Indo-West Pacific,
and Chelyconus, which is restricted to the New World.
Other Fish-Hunting Conus Clades

The Gastridium clade is unusual in the heterogeneity of prey-capture strategies among species. One
of the smaller species, Conus obscurus, appears to use the same general strategy as Pionoconus and
Chelyconus. However, the larger species (Conus geographus and Conus tulipa) use a net strategy;
these species always engulf their fish prey with their false mouths before injecting venom, and they
are believed to specialize primarily in capturing schools of small fish hiding in reef crevices at night.
498 Olivera et al.

C. geographus, in particular, is extremely agile, despite its relatively large size. It has been observed
to secrete a string of mucus that allows it to levitate from the edge of a rock, very much like a spider
spinning silk to lower itself from a tree branch. Thus, it can presumably closely approach prey.
Aquarium observations suggest that C. geographus secretes venom once the snail is close to prey,
causing fish to act in an abnormal manner—they become hypoactive and, ultimately, are unable to
swim away (2). This presumably would make a school of small fish in a reef crevice easier for the
cone snail to engulf. Thus, instead of having a lightning strike cabal, C. geographus apparently has
evolved a set of venom peptides that cause hypoactivity in the fish nervous system (see sidebar, The
Deadliest Snail). Together this set of peptides is referred to as the nirvana cabal (58).
In terms of species numbers, the largest clade of fish-hunting cone snails is Phasmaconus. There
is a general dearth of literature on the biology of this group; it is even possible that not all
Phasmaconus species are piscivorous. There is a brief note in the literature that Conus parius hunts
fish. Recently, Conus flavus was observed attacking a fish; the highly striped proboscis of C. flavus
was extended, and venom was injected by simply stabbing the fish with the radular tooth. All
Phasmaconus species have needlelike, rather than harpoon-shaped and barbed, radular teeth, as
do species in Pionoconus and Chelyconus. Thus, these snails apparently do not use the radular
tooth to tether their fish prey. In this case, after injecting venom, C. flavus simply followed the fish
around until it was apparently immobilized. It then engulfed the fish from the tail end with its false
mouth (rostrum). In one instance, the fish appeared to be tetanically paralyzed but was experi-
encing severe tremors in an immobilized state. Given this very limited set of observations, it would
appear that the basic strategy for prey capture is behaviorally quite divergent in Phasmaconus
compared with the other fish-hunting clades discussed above.
Curiously, the few direct observations on piscivory in the Textilia clade seem to suggest
a similarity to Pionoconus and Chelyconus in prey-capture strategy. Only Conus bullatus has been
observed to capture fish, and it clearly causes excitotoxic shock, suggesting that a lightning strike
cabal is present in the venom of Textilia species as well. However, the major venom components
responsible for prey capture have not been identified.
GENERATING BIODIVERSITY: AN INTEGRATED NATURAL HISTORY OF

CONE SNAILS (CONIDAE)
Molecular Data and the Genesis of Clades

Traditional taxonomy based on shell morphology and a long interest in Conus by malacologists
over several centuries have provided an unusually comprehensive taxonomic literature on fish-
THE DEADLIEST SNAIL
The geography cone, Conus geographus, has been responsible for most of the human fatalities from cone snail
envenomation documented in the literature. In the absence of medical intervention, approximately 70% of the stings
of the geography cone are fatal, a remarkable rate of human lethality for a relatively small animal. A detailed
description of the human stinging case that was most comprehensively recorded in the past century at Hayman
Island, Australia, is provided in Reference 89. Also provided in this reference is a reprint from Nature, describing
how an island culture in the South Pacific clearly came up with an appropriate response to the sting of cone snails.
Traditionally, the shell-collector’s literature refers to “five deadly cones,” but there are no verified reports for the
other four species.

hunting species. Kohn’s (54, 59–61) classical studies not only led to the discovery of piscivory in
Conus but also provided an ecological framework for understanding prey-hunting behavior.
Aquarium and field observations present a picture of diverse behaviors that lead to prey capture by
different cone snails. Nevertheless, the true relationships between different lineages of cone snails
long remained ambiguous.
Initial molecular phylogenetic data first clarified the divergence of clades of Conus, such as
Chelyconus and Pionoconus (25–27). The molecular phylogenetic data alone unambiguously
separate these piscivorous clades, but results of the molecular characterization of venom
components, accompanied by an elucidation of the mechanism of action of many of the major
venom components, provide independent evidence strongly supporting the molecular phylo-
genetic data.
Thus, an unparalleled opportunity exists to dissect how the convergent evolution of two groups
of piscivorous cone snails, Pionoconus and Chelyconus, may have occurred. It should be em-
phasized that although the molecular phylogenetic data are unequivocal with respect to the
divergent origins of these piscivorous lineages, their evolutionary relationship to the various worm-
hunting Conus lineages is less clear. Although the separation of these two clades is consistent with all
phylogenetic data, elucidating the deeper relationships among the Conus clades has been more
difficult. Surprisingly, for these deep splits, the very short intron 9 sequence from the g-glutamyl
carboxylase gene is far more phylogenetically informative than larger conventional genes (e.g., 12-
and 16SrRNAs). It not only unambiguously establishes many of these relationships among Conus
species but indeed clearly separates the Conasprella conids from the Conus (28, 29). Research now in
progress shows a similar use for discerning evolutionary relationships among members of the
Turridae (P. Showers Corneli, M. Watkins, B.M. Olivera, manuscript in preparation).
Molecular results also show that species within a clade have homologous toxins, always
encoded by the same gene superfamily or even the same subfamily (e.g., a-conotoxins; see Ref-
erences 62, 63). Thus, although there is considerable divergence in amino acid sequence, there is
a striking uniformity of functionally homologous toxins within a clade. These data are particularly
rich for Pionoconus. In every case, the principal toxin that targets the nicotinic acetylcholine
receptor muscle subtype, a major target for paralyzing prey, is a subfamily of a-conotoxins called
the a3/5 subfamily (Table 2). In contrast, the major toxins that have the analogous physiological
mechanism in Chelyconus species are the aA-conotoxins (Table 2). However, in both Chelyconus
and Pionoconus species, the toxins that inhibit the inactivation of Naþ channels (and therefore
play a major role in the lightning strike cabal) are very similar, belonging to the d-conotoxin family
(51). Thus, there is a clear divergence in the nicotinic receptor antagonist subfamilies recruited in
Figure 7
→
Contrasting biodiversity of two fish-hunting Conus clades, Pionoconus and Chelyconus. Some of the species that
belong to the subgenus Pionoconus, a fish-hunting clade found in the Indo-Pacific, are shown. (Center) The only
two species known in the Chelyconus clade, the only fish-hunting species in the New World. (Left) Conus
ermineus, Cape Verde Islands; (Right) Conus purpurascens, Gobernadora Island, Western Panama. Note the
similarity between the two species in Chelyconus and the enormous diversity in size, pattern, and color of the shells
of the Pionoconus species. The shells of species in the Pionoconus clade shown are (a) Conus barthelemyi,
Mauritius; (b) Conus gubernator, East Africa; (c) Conus koukae, Oman (previously regarded as ardisaceus
variety); (d) Conus striolatus, Marinduque Island, Philippines; (e) Conus monachus, Marinduque Island,
Philippines; (f) Conus catus, Marquesa; (g) Conus floccatus, Siargao Island, Philippines; (h) Conus fischoederi,
Turtle Islands, Sulu Sea; (i) Conus stercusmuscarum, Cebu Island, Philippines; (j) Conus striatus, Oahu, Hawaii;
(k) Conus nimbosus, Mozambique; (l) Conus circumcisus, Olango Island, Philippines; (m) Conus fulmen, Japan;
(n) Conus achatinus, Marinduque Island, Philippines; (o) Conus consors, Cebu, Philippines; (p) Conus aurisiacus,
Manando, Sulawesi; (q) Conus timorensis, Mauritius; (r) Conus leobottoni, Balabac Island, Philippines; (s) Conus
gauguini, Marquesas; (t) Conus magus, Bantayan Island, Philippines.
500 Olivera et al.

the two clades, but there is much more similarity in the Naþ channel–targeted toxins across the two
clades.
The most dramatic divergence of all occurs with respect to Kþ channel blockers. In Pionoconus,
these are conkunitzins, small proteins containing a well-known, widely distributed protein fold Kunitz domain:
characteristic protein
known as a Kunitz domain. In contrast, the analogous physiological role in Chelyconus is filled by
fold found in secreted
the k-conotoxins (51, 64)—the two groups of peptides are completely unrelated both structurally small proteins with
and genetically and show no apparent amino acid similarity whatsoever. Thus, there is a dramatic three disulfide
crosslinks; most are
protease inhibitors,
and some are toxins
b g
e i
a
t
j
d f
c h
s o
q k
m
1 cm p l

Table 1 a-Conotoxins from the Pionoconus clade
(a) a3/5-Conotoxin subfamily
Conus species Peptide Sequence
achatinus Ac1.3 NGRCCHPACGKHFICGR

aurisiacus A1.1 NGRCCHPACARKYNCGR
catus a-CI NGRCCHPACGKHFSCGR
circumcisus Cr1.1 GRGRCCHPACGPNYSCGR
magus a-MI GRCCHPACGKNYSCG

monachus Mn1.4 NGRCCHPACAKYFSC
striatus a-SIA YCCHPACGKNFDCGR
(b) a-Conotoxins expressed in two different Pionoconus species

Conus circumcisus Conus monachus
Peptide Sequence Peptide Sequence
Cr1.1 GRGRCCHPACGPNYSCGR Mn1.1 NGRCCHPACGKHFSCGR

Cr1.2 GCCSNPVCHVEHPELCRRRR Mn1.2 NGHCCHPACGGKYVKCG
Cr1.3 GCCGNPDCTSHSCD Mn1.3 NGRCCHPACGAKYFKCG
Cr1.4 RGCCSHPVCNLNNPQMCRGRR Mn1.4 NGRCCHPACAKYFSC
Cr1.5 GCCSNPVCHVEHSYMCGRRR Mn1.5 NGRCCHPACGKYFSC
Cr1.6 GGCCSHPVCYFNNPQMCRGRR Mn1.6 GCCSNPACHVDHAELCRRRR

Cr1.7 GGCCSHPVCNLNNPQMCRGRR
(c) Summary of a-conotoxin subfamily distribution
Conus species Total a3/5 / a3/6 a4/7 a4/4
aurisiacus 5 2 2 1
circumcisus 7 1 5 1
stercusmuscarum 5 1 3 1
consors 6 4 2 0
magus 6 5 1 0
monachus 6 5 1 0
contrast between the Naþ channel– and the Kþ channel–targeted peptides that comprise the
lightning strike cabal: In the former case, there is striking conservation between species in Pio-
noconus and Chelyconus, but in the latter case, the toxins could not be more different, with
different gene superfamilies recruited in the two clades.
The striking differences noted for the d-conotoxins and the k-conotoxins/conkunitzins
suggest a specific pathway for the origins of the Pionoconus and Chelyconus clades (see
Figure 8). The conservation of d-conotoxins implies that these peptides were already present in the
presumptive worm-hunting ancestor common to both of these piscivorous lineages. Why would
502 Olivera et al.

Table 2 Comparison of analogous a- and d-conotoxins from Pionoconus and Chelyconus clades
(a) d-Conotoxins from Pionoconus and Chelyconus
aurisiacus A6.5 DGCSNAGAFCGIHPGLCCSEICIVWCT

catus C6.2 YGCSNAGAFCGIHPGLCCSELCLVWCT
circumcisus Cr6.5 WCIPSGDLCFRSDHIQCCSAKCAFVCL
consors Cn6.10 DGCYNAGTFCGIRPGLCCSEFCFLWCITFVDS
magus M6.3 DGCYNAGTFCGIRPGLCCSEFCFLWCITF

monachus Mn6.3 YECYSTGTGCGINGGLCCSNLCLFFVCLTFS
stercusmuscarum Sm6.6 DGCSSGGTFCGIRPGLCCSEFCFLWCITF
striatus dSVIE DGCSSGGTFCGIHPGLCCSEFCFLWCITF

ermineus E6.14 GDCSPGGTFCGIKHGLCCSGFCVMWCTP

purpurascens d-PVIA EACYAPGTFCGIKGPLCCSEFCLPGVCFG
(b) Pionoconus a3/5-conotoxin subfamily, Cheylconus aA toxins
achatinus Ac1.3 NGRCCHPACGKHFICGR

aurisiacus A1.1 NGRCCHPACARKYNCGR
catus a-CI NGRCCHPACGKHFSCGR
circumcisus Cr1.1 GRGRCCHPACGPNYSCGR
magus a-MI GRCCHPACGKNYSCG
monachus Mn1.4 NGRCCHPACAKYFSC

striatus a-SIA YCCHPACGKNFDCGR
ermineus aAEIVA GCCGPYPNAACHPCGCKVGRPPYCDRPSGG
purpurascens aAPIVA GCCSPYPNAACHPCSCKDRPSYCGQ
d-conotoxins that affected fish be present in the venom of a worm-hunting cone snail? Imperial
et al. (65) suggested that fish would compete with cone snails for their worm prey: A passing
fish could try to steal an envenomated worm from the cone snail predator. Thus, to deter
competition for the worm, snails may have recruited d-conotoxins that would have elicited pain
in injected fish.
What precipitated the shift from worm-hunting to fish-hunting behavior (Figure 8)? The
Kþ channel–targeted toxins are the postulated key molecular component: If a Kþ channel–
targeted toxin affected the same axons as the Naþ channel–targeted d-conotoxins, then
a massive depolarization of that axon would result, and the lightning strike syndrome could
lead to total immobilization in the fish. Worm-hunting cone snails that had serendipitously
evolved a toxin with cross-reactivity on the appropriate Kþ channels in fish competitors could
choose to eat the immobilized fish instead of (or in addition to) the worm. This may have
occurred at least two times in the course of evolution, giving rise to an Indo-Pacific clade and
a new-world clade of fish-hunting cone snails. This model implies that the Chelyconus and

a b
Conus radiatus
IV
C. flavus
C. ermineus
III
C. purpurascens
C. figulinus
C. loroisii
C. betulinus
C. magus
C. consors I
C. achatinus
C. striolatus
C. aurisiacus
C. circumciscus
C. stercusmuscarum
C. striatus
C. bullatus
C. obscurus
C. geographus II
?
C. ammiralis
C. textile
C. marmoreus
C. bandanus
C. acutangulus
C. distans
C. californicus
C. memiae
Figure 8
Origins of two fish-hunting clades [adapted from Imperial et al. (65)]. Both an unexpected conservation and divergence of peptide toxin
sequences are found when two clades of fish-hunting cone snails are compared, depending on the peptide toxin family analyzed. (a)
Illustration of how fish hunting may have arisen from a worm-hunting ancestor. (Top) Any snail that devours worms is in jeopardy of
losing its prey to a fish that could steal the worm before the snail has a chance to totally engulf it. A deterrent to such competitors clearly
could have evolved; one possible deterrent in the venom of these snails is the d-conotoxins that would presumably activate sensory
circuitry, causing pain. (Middle) If the snail also had a Kþ channel blocker in its venom that was at least mildly active on fish Kþ channels, the
combination of the d-conotoxin and Kþ channel inhibition could result in immobilization of the envenomated fish. (Bottom) As the
panel shows, Conus species that could achieve this would have two choices: They could ignore the fish and continue eating worms, but some
intrepid individuals might decide to eat the immobilized fish, which could be the first step in the evolution of fish hunting. (b) The above
scenario may have occurred twice; the d-conotoxins are structurally conserved, with considerable amino acid–sequence identity across
species in both Chelyconus (clade III) and Pionoconus (clade I). In contrast, however, the Kþ channel blockers are completely
unrelated—a small polypeptide with a Kunitz domain, called a conkunitzin, is present in clade I, but a conventional 3-disulfide crosslinked
conotoxin, significantly smaller, is the Kþ channel antagonist in clade III (in contrast to the relative conservation of structure and sequence for
the d-conotoxins shown in Table 2). The conservation of d-conotoxins, and the striking divergence in the Kþ channel–targeted toxins
between Pionoconus and Chelyconus, is the basis for the suggestion that fish hunting evolved independently more than once. The hypothetical
Kþ channel blocker in the two ancestral founder species for Chelyconus and Pionoconus is postulated to be from two different gene
superfamilies, the O-conotoxin superfamily (Chelyconus) and the Conkunitzin family (Pionoconus).
Pionoconus clades arose from different worm-hunting clades that had different types of Kþ
channel–targeted toxins.
Mollusc-Hunting Conus Clades

The molluscivorous Conus clades that hunt other marine snails comprise another group of cone
snails that evolved from worm-hunting ancestors. All snail-hunting lineages are found together
in a single major branch of the phylogenetic tree, including, among others, the type species of
the genus Conus, Conus marmoreus. Therefore, in contrast to piscivory, snail hunting likely
arose only once within the genus Conus. A molecular phylogenetic analysis using three mi-
tochondrial genes and one nuclear gene divided molluscivorous Conus into three major clades:
504 Olivera et al.

Conus s.s., Darioconus, and Cylinder (30). All snail-hunting lineages occur exclusively in the
Indo-Pacific, and no snail-hunting Conus species evolved in the Caribbean or the Atlantic
Ocean. The single molluscivorous species in the Panamic region belongs to the Cylinder clade
and is demonstrably a migrant from the Indo-Pacific, descended from ancestral molluscivorous
Indo-Pacific species (24).
Two well-known molluscivorous species are the type species of the genus Conus, C. mar-
moreus, and of the subgenus Cylinder, Conus textile. Both are relatively large species and quite
abundant in the localities where they occur. A problem faced by a predatory Conus that preys on
other snails is that any gastropod, when attacked, retreats into its shell. Even after a predator
successfully injected venom, it could not access the soft body of a snail who retreated into its shell
without a mechanism for breaking the shell. Thus, to successfully colonize soft gastropod molluscs
as prey, a major barrier needed to be overcome: The envenomated prey needed to be immobilized
outside its shell. To achieve this, many molliscivorous cone snails inject potential prey more than
once and apply venom over as broad an anatomical area as possible (66).
In the case of C. textile, venom injection immediately causes a writhing motion in the prey,
which seems to go into a spastic series of alternating contractions and relaxations of its foot
musculature (see Supplemental Video 3). The injected animal is progressively distended further and Supplemental Material
further beyond the protection of its shell. Many prey snails secrete defensive fluids upon injection,
and C. textile has been observed to retreat some distance away from a snail it has just envenomated.
After the injected prey secretes what appears to be a noxious fluid, C. textile uses its foot to fan away
the fluid, after which it approaches the partly immobilized snail for a second venom injection. After
the prey is sufficiently immobilized, C. textile begins to engulf it. By the time the cone snail has
finished devouring the snail, only the empty shell remains, devoid of any soft tissues. To achieve
this, the cone snail must completely relax the columellar muscle attaching the body to the shell.
Other molluscivorous Conus (such as C. marmoreus) appear to cause a progressive relaxation
of the envenomated prey; the spastic behavior elicited by C. textile is not observed after
C. marmoreus envenomation. Physiological strategies for molluscivorous Conus may be di-
ametrically opposite. One strategy could be the equivalent of the lightning strike cabal that
hyperexcites the nervous system of the envenomated prey snail (Conus textile), therefore causing
the spasticity observed after venom injection. The alternative could be the equivalent of the nirvana
cabal (Conus marmoreus), in which the envenomated snail undergoes a progressive relaxation and
hypoactivity of the nervous system.
Worm-Hunting Conus
Most of the known Conus species specialize in hunting polychaete worms, although a few species
envenomate hemichordates (acorn worms), and others prey on eunicids. Among clades that
specialize in different lineages of polychaetes (54, 59, 67, 68), the subgenus Rhizoconus will attack
amphinomid polychaetes (fireworms). Figure 6 shows an example of Conus imperialis attacking
its potential prey. Several clades of vermivorous cone snails specialize in hunting eunicid poly-
chaetes, terebellids, and capitellids (25).
Some vermivorous Conus species are among the largest conoideans, such as Conus betulinus,
which has been observed by divers to eat large polychaetes, in some cases over a meter long. One
diver described a feeding he observed in which the cone snail took more than half an hour to engulf
the entire polychaete.
Californiconus Versus Conus, Divergence Between Two Genera

Within Conidae, the family to which Conus belongs, two basal clades were recently defined
by molecular phylogenetics: Profundiconus and Californiconus (N. Puillandre, P. Bouchet,

T.F. Duda, S. Kauferstein, A.J. Kohn, B.M. Olivera, M. Watkins, C. Meyer, manuscript
submitted). Because all known members of Profundiconus are rare, deep-water species, there is little
literature on this group apart from descriptions of shell morphology and fragmentary molecular
phylogenetic data. Many additional species of Profundiconus likely are yet to be discovered.
In contrast, Californiconus is known from a single species, C. californicus. There is considerable
literature regarding this shallow-water form, which lives off the coast of California and Baja
California. In contrast to all other known species in the Conidae, C. californicus is a generalist
feeder (59). The venoms and anatomy of C. californicus have been investigated (38, 69–79). Thus,
C. californicus is unique in its phylogenetic position in the family Conidae and is one of the most
intensively studied species in the entire superfamily.
Several biological features of C. californicus diverge significantly from those of the species in the
genus Conus. At a macrobiological level, C. californicus is distinctive in that it will attack prey in at
least four different marine phyla (polychaete worms, other molluscs, fish, and crustaceans). No
other species in the family Conidae has such a wide range of prey. Interestingly, C. californicus has
been observed to hunt in packs to bring down bigger prey. Thus, multiple specimens of C. cal-
ifornicus can attack and feed on prey larger than the snails themselves. This pack strategy is not
observed in any other species in the family Conidae, no matter what the prey, suggesting a diversity
in social behavior among conoidean genera.
What can be much more quantitatively characterized is the molecular divergence between
venom peptides. The most striking difference is in venom gene superfamilies expressed in Cal-
iforniconus versus in Conus. Whereas some venom gene superfamilies were already established
in the common ancestor of Conus versus Californiconus, including the I-1, O, T, S, and J-1
superfamilies, other dominant gene superfamilies expressed in the venom ducts of the 500 species
of Conus, such as the A and M superfamilies, are absent from Californiconus (69). Californiconus
also features gene superfamilies that have not been found in any species of Conus. Since Cal-
iforniconus and Conus diverged (in the Eocene or earlier), each lineage subsequently recruited new
gene superfamilies but also retained gene superfamilies already established in the venom ducts of
the last common ancestor of the two genera.
DIVERGENCE BETWEEN CONOIDEAN FAMILIES: CONIDAE VERSUS

TEREBRIDAE AND TURRIDAE
The Auger Snails
Apart from species in the family Conidae, the most familiar of the conoideans are the auger snails
(family Terebridae). Many species of Terebridae are large and abundant in tropical shallow-water
marine environments, and because of their distinctive shells (see Figure 1), molluscan taxonomists
since Linnaeus have addressed the systematics of the family. The auger snails are sand dwellers,
and a significant number of species live in the intertidal zone; however, many species live at depths
greater than 100 m. The distribution of Terebridae is similar to that of Conidae; the vast majority
of species are found in warm, tropical waters, and the greatest biodiversity is found in the Indo-
Pacific.
A significant number of species in Terebridae have lost their venom ducts, notably, in the genus
Oxymeris (such as Oxymeris maculata, the largest species in the Terebridae) and the genus
Myurella. Two major lineages of auger snails are venomous; one of these is the genus Terebra (s.s.).
The type species Terebra subulata is known to live in relatively calm tropical areas and is known to
prey on small, tube-dwelling polychaetes (80). A second large and successful lineage in the family,
typified by species assigned to the genus Hastula, is usually found on tropical, surf-washed
506 Olivera et al.

beaches. Miller (80) recorded the feeding process in Hastula. These snails inject venom through
a hypodermic-type radular tooth, and interestingly, some species complete prey capture and
envenomation between the passages of two successive ocean waves.
One problem with the taxonomy of the Terebridae family is the instability of the generic
assignments. Bratcher & Cernohorsky (13) assigned most species to either the genus Terebra or the
genus Hastula, assigning a relatively minor fraction of species to other genera. In more recent
treatments, there are a far larger number of genera, but the criteria used for generic assignments
remain obscure. Considerable progress has been made in elucidating the phylogeny of Terebridae
by using molecular markers (12, 14–16). The most recent molecular phylogeny data have been
used to map out the appearance of various characters, particularly anatomical structures asso-
ciated with envenomation. The family Terebridae was divided into ten clades (which did not
correspond to generic assignments using the traditional systematics) that can be correlated with the
presence or absence of a proboscis and/or venom gland. Most species in five of the clades had lost
both the proboscis and the venom gland, whereas both were mostly present in species from the
other five clades, suggesting that the latter use venom for prey capture.
The present molecular information on venom composition in Terebridae is rudimentary; data

are available for only three species. The most extensively analyzed is Hastula (Impages) hectica.
Sequences of 15 venom peptides, 8 of which were purified directly from venom, have been reported
for this species (15). The biochemical properties of the 15 predicted peptide toxins were generally
similar to those of Conus venoms: Most were relatively short peptides, under 40 AA in length,
with multiple disulfide bonds. There are two noteworthy differences. The frequency of post-
translational modifications appears to be significantly lower in the venom peptides of H. hectica
compared with those of most Conus species. Furthermore, there is no detectable overlap in the gene
superfamilies expressed in H. hectica and any Conus species; however, one of the peptides showed
significant sequence similarity to a peptide from the family Turridae, Lophiotoma olangoensis.
Thus, although the family Terebridae has some general similarities to Conidae, significant
differences appear even in the very preliminary results available so far. It is notable that such a high
proportion of the species have apparently lost their venom apparatus; as far as is known, all of the
700 species in Conidae have a venom gland. The molecular phylogeny suggests that the loss of the
venom apparatus has occurred multiple times in the evolutionary history of Terebridae.
The Family Turridae

As classically defined, the family Turridae includes all conoidean species that are neither cone
snails nor auger snails. In more recent systematic treatments, the number of genera included in the
family Turridae has been progressively restricted to a smaller subset. Recent authors (4, 17, 40, 81)
have restricted the family Turridae to the genera that Powell included in the subfamily Turrinae
(82). After the 358 former turrid genera and subgenera have been attributed to 13 apparently
monophyletic families, the newly redefined family Turridae includes only 16 genera.
Even in this reduced form, the family Turridae presents highly confused systematics. The
proposed genera and subgenera within the redefined Turridae do not correspond to monophyletic
branches of the phylogenetic tree, and a new generic framework is needed. For example, the genus
Turris (35, 82) is clearly split into two major groups of species, one more closely related to the
species traditionally assigned to other genera (e.g., Gemmula or Lophiotoma) than to the other
group of species generally assigned to Turris (40). Many turrid genera, especially Gemmula and
Unedogemmula, combine sets of unrelated lineages and encompass a high proportion of cryptic
species, distinctive based on molecular data but indistinguishable morphologically (N. Puillandre,
P. Bouchet, T.F. Duda, S. Kauferstein, A.J. Kohn, B.M. Olivera, M. Watkins, C. Meyer,

manuscript submitted). Thus, the systematics of the family and the present molecular phylogenetic
data do not correspond to each other.
The shell morphology of species in the family Turridae is distinctive; many of the larger forms
have long siphonal canals, and the shape of the shells in the family is generally narrowly fusiform.
Surprisingly little is known about their biology; Heralde et al. (81) established recently that several
species of the Turridae prey on terebellid polychaetes. In a recent video recording of feeding by
a species of Turris, envenomation occurred while the predator was buried beneath the substrate.
The presence of a shelly siphonal canal apparently allows the animal to maneuver its proboscis
toward the polychaete prey under the substrate and successfully envenomate the prey. This is
clearly quite different from envenomation by species in Conidae or Terebridae. Envenomation in
the terebellid worm causes spastic, uncoordinated activity, quite analogous to the hyperactivation
of the fish nervous system through the lightning strike cabal of Pionoconus species and of
molluscivorous Conus in the subgenus Cylinder (58).
Peptide toxins from the venoms of several members of Turridae have been characterized, either
by transcriptome analysis or by direct purification from venom (83–86). This work has already
revealed that some of the more specialized posttranslational modifications found in Conus venom
peptides (e.g., g-carboxylation of glutamate, bromination of tryptophan) also occur in venom
peptides of turrids (in contrast to the observations of Terebridae noted above).
PERSPECTIVES ON THE BIODIVERSITY OF VENOMOUS MARINE SNAILS

The origin of venomous marine molluscs can be traced back to the Cretaceous; by the early
Tertiary period, the various families were probably established. It is clear that each family group
within Conoidea largely recruited its own set of gene superfamilies to be expressed in venom. A
combination of the molecular data for the Turridae, Terebridae, and Conidae shows that there is
little overlap in the venom gene superfamilies recruited in the different conoidean family groups.
So far, only one of the twelve conotoxin superfamilies is shared between the Turridae and Conidae,
and another gene superfamily is shared between the Turridae and Terebridae. These gene
superfamilies were presumably already recruited for expression in venom in the ancestral species,
giving rise to the family groups. At present, the physiological mechanisms that underlie the ac-
tivities of peptides in these gene superfamilies are unknown; the elucidation of their mechanisms
may provide insight into how the original ancestors used their venoms.
There is greater overlap between the gene superfamilies recruited when two different genera are
compared within the same family group. The best data are for Californiconus versus Conus in the
family Conidae, where approximately half of the gene superfamilies were presumably present in
the venom of the last common ancestor, but each lineage later recruited additional gene super-
families after divergence of the two genera. Within a single genus, Conus, most gene superfamilies
are shared across species, but different clades have selectively evolved more members of selected
superfamilies. The distribution of venom peptides in each gene superfamily varies considerably
between the different Conus clades and, sometimes, even within species in the same clade (87). The
gene superfamilies then undergo various specializations within a clade, which may be charac-
teristic of only a small number of closely related species.
Clearly, past and recent studies of the extremely speciose venomous marine snails have pro-
vided impressive evidence for the extraordinary biodiversity of the Conoidea. But as we mentioned
earlier, the low level of research allotted to the vast majority of the >10,000 conoid species, and the
concentration of multidisciplinary studies on a relatively miniscule fraction belonging to the
Conidae, means that a great deal of work remains to be done. Fortunately, the conid research
provides mechanistic insight to guide the progress of research for the understudied terebrids and
508 Olivera et al.

turrids by showing how diverse strategies have evolved within the highly successful genus Conus. A
combination of molecular phylogeny, toxinology, behavioral observations, and traditional sys-
tematics suggests how fish hunting evolved independently in Conus at two different geographic
locations early in the Miocene. The goal is to more fully research other lineages of Conidae, the
Terebridae, and most of all the taxonomically unruly Turridae to reveal the true scope of bio-
diversity in the Conoidea.
DISCLOSURE STATEMENT
The authors are not aware of any affiliations, memberships, funding, or financial holdings that
might be perceived as affecting the objectivity of this review.
ACKNOWLEDGMENTS
The authors’ research has been supported primarily by a grant from the National Institute of
General Medical Sciences, GM48677. The figures, specimens, and videos were made in conjunc-
tion with research supported by ICBG Grant 1U01TW008163 from the Fogarty Center, National
Institutes of Health, and outreach activities supported by a Professors Award from the Howard
Hughes Medical Institute to B.M.O. We are deeply grateful to Terry Merritt and My Hyunh, who
were our partners in assembling this review.
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85. López-Vera E, Heimer de la Cotera EP, Maillo M, Riesgo-Escovar JR, Olivera BM, Aguilar MB. 2004. A
novel structural class of toxins: the methionine-rich peptides from the venoms of turrid marine snails
(Mollusca, Conoidea). Toxicon 43:365–74
86. Watkins M, Hillyard DR, Olivera BM. 2006. Genes expressed in a turrid venom duct: divergence and
similarity to conotoxins. J. Mol. Evol. 62:247–56
87. Lluisma AO, Milash BA, Moore B, Olivera BM, Bandyopadhyay P. 2012. Novel venom peptides from the
cone snail Conus pulicarius discovered through next-generation sequencing of its venom duct tran-
scriptome. Mar. Genomics 5:43–51
88. Miljanich GP. 2004. Ziconotide: neuronal calcium channel blocker for treating severe chronic pain. Curr.
Med. Chem. 11:3029–40
89. Flecker H. 1936. Cone shell mollusc poisoning, with report of a fatal case. Med. J. Aust. 1:464–66
512 Olivera et al.

RELATED RESOURCES
WoRMS (World Register of Marine Species). A comprehensive, taxonomist-directed list of
marine organisms with current and historical taxonomy and synonomy. Links to other
valuable sites. http://www.marinespecies.org
The Cone Snail: Olivera Lab at the University of Utah. Photos, videos, evolution, taxonomy,
hunting behavior, venoms, adaptations. http://www.theconesnail.com
The ConoServer: Database resource for conopeptide protein, nucleotides, and 3-D structure.
http://www.conoserver.org
Coneshell.Net Collections: Several hundred beautiful photographs of many species, bibliog-
raphy, and links to other sites. http://www.coneshell.net
Conchylomania: A scholarly webpage describing the relationship of cone shell collections to art
in the context of seventeenth- and eighteenth-century Europe. http://penelope.uchicago.edu/
∼grout/encyclopaedia_romana/aconite/conchylomania.html
Conus geographus: A webpage describing human envenomation by Conus geographus.

http://penelope.uchicago.edu/∼grout/encyclopaedia_romana/aconite/geographus.html

Annual Review of
Animal Biosciences
Volume 2, 2014 Contents
From Germ Cell Preservation to Regenerative Medicine: An Exciting

Research Career in Biotechnology

Ian Wilmut . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Genomic Imprinting in Farm Animals

Xiuchun (Cindy) Tian . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Recent Advances in Primate Phylogenomics
Jill Pecon-Slattery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Domestication Genomics: Evidence from Animals
Guo-Dong Wang, Hai-Bing Xie, Min-Sheng Peng, David Irwin,
and Ya-Ping Zhang . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Behavior Genetics and the Domestication of Animals
Per Jensen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Applied Animal Genomics: Results from the Field
Alison L. Van Eenennaam, Kent A. Weigel, Amy E. Young,
Matthew A. Cleveland, and Jack C.M. Dekkers . . . . . . . . . . . . . . . . . . . . 105
Pestiviruses
Matthias Schweizer and Ernst Peterhans . . . . . . . . . . . . . . . . . . . . . . . . . 141
Pathogenesis and Molecular Biology of a Transmissible Tumor in the
Tasmanian Devil
Hannah S. Bender, Jennifer A. Marshall Graves, and Janine E. Deakin . . . 165
Animal Models of Bovine Leukemia Virus and Human T-Lymphotrophic
Virus Type-1: Insights in Transmission and Pathogenesis
Michael D. Lairmore . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
Malignant Catarrhal Fever: Inching Toward Understanding
Hong Li, Cristina W. Cunha, Naomi S. Taus, and Donald P. Knowles . . . 209
vi
Comparative Immune Systems in Animals
Shaochun Yuan, Xin Tao, Shengfeng Huang, Shangwu Chen,
and Anlong Xu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235
Origin and Evolution of Adaptive Immunity
Thomas Boehm and Jeremy B. Swann . . . . . . . . . . . . . . . . . . . . . . . . . . . 259
The Functional Significance of Cattle Major Histocompatibility Complex
Class I Genetic Diversity
Shirley A. Ellis and John A. Hammond . . . . . . . . . . . . . . . . . . . . . . . . . . 285
Incidence of Abnormal Offspring from Cloning and Other Assisted
Reproductive Technologies
Jonathan R. Hill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307

Preadipocyte and Adipose Tissue Differentiation in Meat Animals:
Influence of Species and Anatomical Location

G.J. Hausman, U. Basu, S. Wei, D.B. Hausman, and M.V. Dodson . . . . . 323
Serotonin: A Local Regulator in the Mammary Gland Epithelium
Nelson D. Horseman and Robert J. Collier . . . . . . . . . . . . . . . . . . . . . . . 353
Evolution of the Modern Broiler and Feed Efficiency
Paul B. Siegel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 375
Amino Acid Nutrition in Animals: Protein Synthesis and Beyond
Guoyao Wu, Fuller W. Bazer, Zhaolai Dai, Defa Li, Junjun Wang,
and Zhenlong Wu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 387
The Suckling Piglet as an Agrimedical Model for the Study of Pediatric
Nutrition and Metabolism
Jack Odle, Xi Lin, Sheila K. Jacobi, Sung Woo Kim,
and Chad H. Stahl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 419
Cattle Production Systems: Ecology of Existing and Emerging
Escherichia coli Types Related to Foodborne Illness
David R. Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445
Gastrointestinal Tract Microbiota and Probiotics in Production Animals
Carl J. Yeoman and Bryan A. White . . . . . . . . . . . . . . . . . . . . . . . . . . . . 469
Biodiversity of Cone Snails and Other Venomous Marine Gastropods:
Evolutionary Success Through Neuropharmacology
Baldomero M. Olivera, Patrice Showers Corneli, Maren Watkins,
and Alexander Fedosov . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 487
Ecological Risk Analysis and Genetically Modified Salmon: Management
in the Face of Uncertainty
Darek T.R. Moreau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 515
Contents vii
The Modern Feedlot for Finishing Cattle
John J. Wagner, Shawn L. Archibeque, and Dillon M. Feuz . . . . . . . . . . . 535
The Nexus of Environmental Quality and Livestock Welfare
Sara E. Place and Frank M. Mitloehner . . . . . . . . . . . . . . . . . . . . . . . . . . 555
Errata
An online log of corrections to Annual Review of Animal Biosciences articles may

be found at http://www.annualreviews.org/errata/animal
viii Contents
Annual Reviews
It’s about time. Your time. It’s time well spent.
New From Annual Reviews:

Annual Review of Statistics and Its Application
Volume 1 • Online January 2014 • http://statistics.annualreviews.org
Editor: Stephen E. Fienberg, Carnegie Mellon University

Associate Editors: Nancy Reid, University of Toronto
Stephen M. Stigler, University of Chicago
The Annual Review of Statistics and Its Application aims to inform statisticians and quantitative methodologists, as
well as all scientists and users of statistics about major methodological advances and the computational tools that
allow for their implementation. It will include developments in the field of statistics, including theoretical statistical
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and bioinformatics, economics, machine learning, psychology, sociology, and aspects of the physical sciences.
Complimentary online access to the first volume will be available until January 2015.
table of contents:
• What Is Statistics? Stephen E. Fienberg • High-Dimensional Statistics with a View Toward Applications
• A Systematic Statistical Approach to Evaluating Evidence in Biology, Peter Bühlmann, Markus Kalisch, Lukas Meier
from Observational Studies, David Madigan, Paul E. Stang, • Next-Generation Statistical Genetics: Modeling, Penalization,
Jesse A. Berlin, Martijn Schuemie, J. Marc Overhage, and Optimization in High-Dimensional Data, Kenneth Lange,
Marc A. Suchard, Bill Dumouchel, Abraham G. Hartzema, Jeanette C. Papp, Janet S. Sinsheimer, Eric M. Sobel
Patrick B. Ryan • Breaking Bad: Two Decades of Life-Course Data Analysis
• The Role of Statistics in the Discovery of a Higgs Boson, in Criminology, Developmental Psychology, and Beyond,
David A. van Dyk Elena A. Erosheva, Ross L. Matsueda, Donatello Telesca
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• Statistics and Climate, Peter Guttorp • Statistical Evaluation of Forensic DNA Profile Evidence,
• Climate Simulators and Climate Projections, Christopher D. Steele, David J. Balding
Jonathan Rougier, Michael Goldstein • Using League Table Rankings in Public Policy Formation:
• Probabilistic Forecasting, Tilmann Gneiting, Statistical Issues, Harvey Goldstein
Matthias Katzfuss • Statistical Ecology, Ruth King
• Bayesian Computational Tools, Christian P. Robert • Estimating the Number of Species in Microbial Diversity
• Bayesian Computation Via Markov Chain Monte Carlo, Studies, John Bunge, Amy Willis, Fiona Walsh
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• Build, Compute, Critique, Repeat: Data Analysis with Latent Susan A. Murphy
Variable Models, David M. Blei • Statistics and Related Topics in Single-Molecule Biophysics,
• Structured Regularizers for High-Dimensional Problems: Hong Qian, S.C. Kou
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Biodiversity of Cone Snails and Other Venomous Marine Gastropods:

Article in Annual Review of Animal Biosciences · February 2014

Baldomero Olivera Patrice Showers Corneli

SEE PROFILE SEE PROFILE

Phylogenomics of the Neogastropoda View project

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Annu. Rev. Anim. Biosci. 2014. 2:487–513 Keywords

The Annual Review of Animal Biosciences is online Abstract

2, 8, 9). Furthermore, because of the neuropharmacological interest in their venom components

SYSTEMATICS AND PHYLOGENY OF CONOIDEA

CONE SHELLS: TREASURES AND ART OBJECTS

488 Olivera et al.

PRESENT AND FUTURE BIOMEDICAL APPLICATIONS OF MARINE SNAIL VENOMS

www.annualreviews.org Venomous Marine Snails 489

Conus imperialis Conus marmoreus g

Cone Snails: Recent Advances in Taxonomy and Phylogeny

490 Olivera et al.

www.annualreviews.org Venomous Marine Snails 491

492 Olivera et al.

www.annualreviews.org Venomous Marine Snails 493

Molecular Physiology of Envenomation

494 Olivera et al.

DIVERSITY OF THE FISH-HUNTING CONE SNAIL CLADES

The Pionoconus Clade of Fish-Hunting Conus: Similarity and Divergence of Species

www.annualreviews.org Venomous Marine Snails 495

496 Olivera et al.

Two Fish-Hunting Clades, Chelyconus and Pionoconus: Convergent Evolution

www.annualreviews.org Venomous Marine Snails 497

Other Fish-Hunting Conus Clades

498 Olivera et al.

GENERATING BIODIVERSITY: AN INTEGRATED NATURAL HISTORY OF

Molecular Data and the Genesis of Clades

THE DEADLIEST SNAIL

www.annualreviews.org Venomous Marine Snails 499

500 Olivera et al.

www.annualreviews.org Venomous Marine Snails 501

(a) a3/5-Conotoxin subfamily

Conus species Peptide Sequence

achatinus Ac1.3 NGRCCHPACGKHFICGR

magus a-MI GRCCHPACGKNYSCG

(b) a-Conotoxins expressed in two different Pionoconus species

Conus circumcisus Conus monachus

Peptide Sequence Peptide Sequence

Cr1.1 GRGRCCHPACGPNYSCGR Mn1.1 NGRCCHPACGKHFSCGR

Cr1.6 GGCCSHPVCYFNNPQMCRGRR Mn1.6 GCCSNPACHVDHAELCRRRR

(c) Summary of a-conotoxin subfamily distribution

Conus species Total a3/5 / a3/6 a4/7 a4/4

502 Olivera et al.

(a) d-Conotoxins from Pionoconus and Chelyconus

Conus species Peptide Sequence

aurisiacus A6.5 DGCSNAGAFCGIHPGLCCSEICIVWCT

magus M6.3 DGCYNAGTFCGIRPGLCCSEFCFLWCITF

striatus dSVIE DGCSSGGTFCGIHPGLCCSEFCFLWCITF

ermineus E6.14 GDCSPGGTFCGIKHGLCCSGFCVMWCTP

(b) Pionoconus a3/5-conotoxin subfamily, Cheylconus aA toxins

Conus species Peptide Sequence

achatinus Ac1.3 NGRCCHPACGKHFICGR

monachus Mn1.4 NGRCCHPACAKYFSC

www.annualreviews.org Venomous Marine Snails 503

Mollusc-Hunting Conus Clades

504 Olivera et al.

Californiconus Versus Conus, Divergence Between Two Genera

Editor: Stephen E. Fienberg, Carnegie Mellon University