Yap Et Al 2023 Taxonomy and Molecular Phylogeny of The Sea Anemone Macrodactyla With A Description of A New Species From Singapore

Zoological Studies 62:29 (2023)
doi:10.6620/ZS.2023.62-29
Open Access
Taxonomy and Molecular Phylogeny of the

Sea Anemone Macrodactyla (Haddon, 1898)
(Cnidaria, Actiniaria), with a Description of a
New Species from Singapore
Nicholas Wei Liang Yap1,2,* , Michela Lee Mitchell3 , Zheng Bin Randolph Quek4 , Ria Tan5,
Koh Siang Tan1,2 , and Danwei Huang1,4,5
1
Tropical Marine Science Institute, National University of Singapore, S2S Building, 18 Kent Ridge Road, Singapore 119227.
Correspondence: E-mail: [email protected] (Yap).
E-mail: [email protected] (Tan); [email protected] (Huang)
2
St. John’s Island National Marine Laboratory, c/o Tropical Marine Science Institute, National University of Singapore, 18 Kent Ridge Road,
Singapore 119227
3
Biodiversity and Geosciences, Museum of Tropical Queensland, 70-102 Flinders Street, Townsville, Queensland, Australia.
E-mail: [email protected] (Mitchell)
4
Department of Biological Sciences, National University of Singapore, Block S3 Level 4, 16 Science Drive 4, Singapore 117558.
E-mail: [email protected] (Quek)
5
c/o Lee Kong Chian Natural History Museum, National University of Singapore, 2 Conservatory Drive, Singapore 117377.
E-mail: [email protected] (Tan)
Received 12 August 2022 / Accepted 28 April 2023 / Published 6 July 2023

Communicated by James D. Reimer
Sea anemones (Cnidaria, Actiniaria) are a successful group of marine invertebrates found in a diverse
range of environments globally. In spite of their ubiquity, identities for many sea anemones remain
unverified, especially those from the Indo-West Pacific region. Here, we clarify the taxonomy of the poorly
known Macrodactyla aspera, a shallow-water species first described from the Torres Straits in northern
Australia. We re-describe M. aspera based on new morphological and molecular data gathered from the
type specimen, other museum vouchers, and from fresh material collected from Singapore. We tested the
monophyly of Macrodactyla using three mitochondrial (12S, 16S and cox3) and one nuclear (28S) marker
based on three congeners, recovering this genus to be polyphyletic. As a consequence, we transferred M.
doreensis to the genus Heteractis, and describe a new species, Macrodactyla fautinae sp. nov. While both
M. aspera and M. fautinae sp. nov. share the same arrangement and number of complete mesenteries, a
similar distribution of cnidae, and are not symbiotically associated with any other biota, M. fautinae sp. nov.
has perforated, lobe-like verrucae on its column, and lacks nematocyst batteries on its tentacles, unlike M.
aspera. These two species also occur in similar habitats in Singapore. Finally, because M. aspera strongly
resembles Dofleinia armata, the latter species flagged as a danger to public health due to its ability to
inflict painful stings, we tested the relationship between these species and found them not to be closely
related. However, tentacles of M. aspera, like D. armata, are densely covered with nematocyst batteries
and harbour large nematocysts; we infer that M. aspera may also be capable of delivering stings that
endanger public health. This study builds upon a growing number of studies that aim to ascertain identities
and systematics of sea anemones historically reported from the Indo-West Pacific. Our findings will
facilitate accurate species identification, which is crucial for advancing research, formulating conservation
measures, and protecting public health.
Key words: Actinioidea, Anthozoa, Intertidal, Integrative taxonomy, Southeast Asia.
Citation: Yap NWL, Mitchell ML, Quek ZBR, Tan R, Tan KS, Huang D. 2023. Taxonomy and molecular phylogeny of the sea anemone Macrodactyla
(Haddon, 1898) (Cnidaria, Actiniaria), with a description of a new species from Singapore. Zool Stud 62:29. doi:10.6620/ZS.2023.62-29.
© 2023 Academia Sinica, Taiwan 1

Zoological Studies 62:29 (2023) page 2 of 25
BACKGROUND greatly impedes further research on these animals.

Here, we focused on clarifying the taxonomy of
Sea anemones (Cnidaria, Actiniaria) are a one Indo-West Pacific species: Macrodactyla aspera
cosmopolitan group of marine invertebrates, occupying (Haddon and Shackleton, 1893). This species is of
a diverse range of habitats globally, extending from the particular interest as it bears a strong resemblance to D.
tropical shallow seas to deep oceanic trenches (Gomes armata, and occurs in similar, sandy and silty habitats
et al. 2016). These animals are engaged in important (Erhardt and Knop 2005; Rowlett 2020). Like D.
marine ecological processes, including symbiotic armata, M. aspera also bears tentacles that are densely
relationships with micro- and macro-organisms (e.g., covered with visible batteries that are packed with
fishes; crustaceans, and dinoflagellates, etc.; see: nematocysts (i.e., nematocyst batteries) which may pose
Dunn 1981, Fransen 1989; Sunagawa et al. 2009, a threat to public health.
respectively). Sea anemones are also sought after in Macrodactyla aspera was first described from
the marine aquarium trade, and in some cultures, these northern Australia by Haddon and Shackleton (1893)
animals are consumed as food (Dunn 1981; Scott et al. as Condylactis aspera (it is also the type species for the
2014). Some are also feared by humans because of their genus Macrodactyla; see Carlgren 1949; Fautin 2016).
ability to inflict painful stings upon contact. Like all Since then, little taxonomic work has been conducted
other cnidarians, sea anemones manufacture intracellular to confirm its identity. Presently, only two published
stinging capsules (known as nematocysts) containing descriptions have dealt directly with the taxonomy of
venom (Fautin 1988 2009; Mitchell et al. 2020). Stings this species: i) Haddon and Shackleton (1893), and
of some tropical shallow-water species from the Indo- ii) Haddon (1898). By contemporary standards, these
West Pacific region may leave severe, long-lasting scars two taxonomic publications lacked crucial taxonomic
on their human victims, or in some instances, cause details to accurately identify the animal (e.g., absence
death (Erhardt and Knop 2005). Amongst species from of cnidom data). This species was most recently
the region that harbour potent stings, and are a threat to referred to in an annotated checklist of littoral biota
public health, four are well-known: Dofleinia armata from the Gulf of Aqaba (Fishelson 1970). A half-
Wassilief, 1908, Phyllodiscus semoni Kwietniewski, tone photograph of the living animal with its tentacles
1897, and members of the family Actinodendridae extended accompanied this checklist (see Fishelson
Haddon, 1898 such as Actinodendrum arboreum (Quoy 1970: 100, Fig. 9), but no taxonomic description was
and Gaimard, 1833) and Actinostephanus haeckeli given. Much of the animal remained obscured in this
Kwietniewski, 1897 (see: Coleman 1999; Hoeksema half-tone photograph – it is unclear what M. aspera
and Crowther 2011; Mizuno et al. 2007; Rowlett 2020). truly looks like when alive. With the exception of
Despite their diversity, commercial popularity, and Fishelson’s (1970) account, it has been more than a
the possible threats they may impose on human health, century since the species boundaries of M. aspera have
identities for many sea anemone species remain poorly been evaluated and tested; this warrants an investigation
verified. This is especially so for those that occur within of the diagnostic features that were previously reported
the Indo-West Pacific region (den Hartog 1997; Fautin et to define this species.
al. 2009; Yap et al. 2019). This taxonomic inadequacy is Other taxonomic works referring to M. aspera
a result of these invertebrates being notoriously difficult were dedicated to revising the original diagnosis of the
to identify accurately with confidence. Sea anemones genus Macrodactyla Haddon, 1898 (i.e., Stephenson
are skeleton-less and among species, they have a 1921 1922; Carlgren 1949; Dunn 1981). This genus was
similar, uniform body-plan and an apparent “dearth of established by Haddon (1898: 431) to accommodate
features” to distinguish them (Fautin 1988: 497). Many C. aspera, as he recognised that it was different from
morphological characters that were historically used known members of Condylactis, although he was
to delimit species boundaries are still poorly defined, not explicit in stating his reasons for transferring this
leading to taxonomic confusion (see Daly 2003; Daly species to Macrodactyla. While Haddon and Shackleton
et al. 2017). Furthermore, emerging molecular evidence (1893) placed this species in Actiniidae Rafinesque,
has revealed that many distantly related species may 1815 – a taxonomic decision retained in Haddon
exhibit a great degree of morphological convergence (1898) and thereafter – the diagnosis of Macrodactyla
among them (Rodríguez et al. 2014; Daly et al. 2017; by Haddon (1898) contradicted accepted diagnoses
Titus et al. 2019). Apart from these aforementioned of Actiniidae during his time. Haddon (1898) had
challenges, many species originally described from the erroneously reported that all mesenteries present in C.
Indo-Pacific region have not been verified, due to a lack aspera were ‘imperfect’ (i.e., incomplete). Stephenson
of taxonomic detail in their original descriptions (Dunn (1922: 263) subsequently flagged Haddon’s account
1974; Fautin et al. 2009; Yap et al. 2019 2021). This as an ‘ambiguity’, since members of Actiniidae were
© 2023 Academia Sinica, Taiwan

accepted at that time to have more than six complete to the use of the name Macrodactyla. This genus name
mesenteries (e.g., Hertwig 1882; Carlgren 1893; was reported to be a junior homonym to a genus of
McMurrich 1889; see: Haddon 1898: 414). After a beetle described in Hitchcock (1833) (see: Neave
examining the type specimen of C. aspera, Stephenson 1940; Fautin 2016). Following Neave’s (1940) listing
(1921 1922) corrected Haddon’s (1898) diagnosis for and strict stipulations outlined by the International
Macrodactyla, confirming the presence of six pairs of Commission on Zoological Nomenclature (i.e., Article
complete mesenteries in the type material. However, 23.3.5; the “Code”, henceforth), Fautin (2016) returned
because Stephenson (1922) had considered actiniids to both species, presently placed in Macrodactyla, to
possess more than six complete pairs of mesenteries, he Condylactis (therefore, as C. aspera and C. doreensis,
moved Macrodactyla to a new family Myonanthidae, respectively). Despite her nomenclatural actions, Fautin
whose members strictly possessed only six complete (2016: 105) also conceded that she had not consulted
pairs. While Carlgren’s (1949) monograph that followed the original publication where the senior homonym had
retained much of Stephenson’s (1921 1922) revised appeared (i.e., Hitchcock 1833), stating, “… not seen:
diagnosis, he returned Macrodactyla to Actiniidae, as “Rep. Geo. Min. Zool. Massach., 575…” (emphasis in
he posited that actiniids have “…Perfect [= complete] bold, our own). Subsequent publications after Fautin
pairs of mesenteries rarely six, as a rule more than six,” (2016) have retained the use of Macrodactyla when
(Carlgren 1949: 47; additions in square parentheses, our referring to both M. aspera and M. doreensis (e.g., Titus
own). et al. 2019; Yap et al. 2021), because their morphology
Present perspectives on the systematics of was not congruent with Condylactis (e.g., presence
Macrodactyla stem from Dunn’s (1981) [= DG of a sphincter muscle; see: González-Muñoz et al.
Fautin] expanded definition of the genus. This genus 2012). The nomenclatural status for the genus name
had remained monotypic since its publication until Macrodactyla needs to be resolved; Neave’s (1940)
Dunn’s (1981) inclusion of a congener: Macrodactyla assertions and Fautin’s (2016) resultant nomenclatural
doreensis (Quoy and Gaimard, 1833). This congener’s actions must be verified for the unambiguous usage of
name was a new combination that Dunn (1981) had this genus name for sea anemones.
created, to apply to a number of sea anemones she In this study, we re-described M. aspera based on
had previously synonymised into one species. Among new data gathered from its type specimen, accompanied
those synonymised were Actinia doreensis Quoy by fresh material collected from Singapore, and from
and Gaimard, 1833 and Condylactis gelam Haddon vouchers kept in natural history museums worldwide.
and Shackleton, 1893 (see Dunn 1981: 29, for the Nomenclatural issues relating to the use of the genus
full synonymy list). While Dunn (1981) was at first name Macrodactyla, were resolved to support its
uncertain in finding an appropriate genus to classify continued usage for sea anemones. We also tested the
these synonymised species, she eventually settled on phylogeny of the two presently accepted species in
Macrodactyla, rationalizing that they were (Dunn 1981: Macrodactyla (i.e., M. aspera and M. doreensis) and
29), “… no doubt closely related [witness Haddon and recovered them as polyphyletic. Considering this new
Shackleton’s (1893) original description of both as evidence, we revised the definition of Macrodactyla,
members of Condylactis]…”. However, Dunn (1981: and classified M. doreensis in Heteractis as Heteractis
35) also noted that name-bearing specimens of these doreensis (Quoy and Gaimard, 1833) comb. nov.
synonymised species (e.g., syntypes of C. gelam), and Additionally, we tested the relationship between M.
the type specimen of the type species for Macrodactyla aspera and D. armata, due to the strong morphological
(i.e., C. aspera) were morphologically different in resemblance between these two species. We found them
the “…distribution and size of cnidae, character of to be distantly related, with D. armata being recovered
mesenterial retractor muscles… and tentacle number with Actinodendridae Haddon, 1898, its members well-
and form,” casting doubt on her own decision. At known to inflict painful stings.
present, it is unclear if M. aspera and M. doreensis are During field surveys conducted for this study
congeneric. Recent phylogenetic analyses using only in Singapore, we encountered another species of sea
M. doreensis samples have repeatedly recovered this anemone that co-occurs with M. aspera. While this
species to be nested within Heteractis Milne Edwards, second species has been photographed and is known by
1857 (Titus et al. 2019; Yap et al. 2021). Prior to this local citizen scientists for many years, being nicknamed
study, phylogenetic interpretation using M. aspera as the ‘Purple-lip Sand Anemone’ and ‘Tiger Anemone’
have not been conducted, and the monophyly of in both printed and web-based biodiversity guides (e.g.,
Macrodactyla remains untested. Rowlett 2020: 323; WildSingapore Fact Sheets: http://
Apart from these systematic problems, Fautin www.wildsingapore.com/wildfacts/, respectively), its
(2016) also highlighted a nomenclatural issue relating identity has remained undetermined until now. This

second species is new to science, which we formally were viewed at 1000x magnification under a light
described as Macrodactyla fautinae sp. nov., and is thus microscope. Only intact capsules (i.e., undischarged
far only known from Singapore. In our phylogenetic capsules) were measured using conventions presented
analyses, we recovered this new species and M. aspera in Dunn (1981) and Yap et al. (2021). Cnidae taxonomy
as a clade. Macrodactyla fautinae sp. nov. differs from followed Mariscal (1974).
M. aspera in bearing adhesive and extensible, perforated Translations of original taxonomic descriptions
lobe-like verrucae, and a lack of visible nematocyst written in languages other than English (e.g., Wassilieff
batteries on its tentacles. 1908) were carried out using Google Translate (Google
Information from this study builds on efforts to Translate: https://translate.google.ca). For specialised
accurately identify sea anemones from the Indo-West scientific terms that could not be translated suitably, we
Pacific region (e.g., England 1987; Fautin et al. 2009; followed definitions listed in Stachowitsch (1992).
Yap et al. 2019). This is necessary to prime future To redefine the species boundaries of M. aspera,
research involving these marine invertebrates from the syntypes of C. aspera housed at both University
region as so little is known about their ecology (e.g., Museum of Zoology, Cambridge, United Kingdom
reproductive biology, see Yong et al. 2021). Moreover, (MZC), and Museum of Zoology, Lund University,
given that sea anemones can impact humans due to their Sweden (MZL), were examined (for type material
ability to deliver harmful stings, precise identification is listing, see: Fautin 2016). Furthermore, we located
key to constructive management and mitigative efforts and examined museum voucher material that closely
in safeguarding public health. resembled M. aspera, or those that were identified as D.
armata on their labels. These vouchers originated from
Australia, New Caledonia, Singapore and Israel (Fig.
MATERIALS AND METHODS S2), in the collections of Muséum National d’Histoire
Naturelle, Paris, France (MNHN), Steinhardt Museum
Sea anemone collection and processing of Natural History, Tel Aviv, Israel (SMNHTAU), and
Western Australian Museum, Perth, Australia (WAM).
Fresh material examined in this study were All fresh material collected from Singapore for
collected from intertidal habitats along Singapore’s this study were deposited into the Zoological Reference
coastlines and offshore islands (Fig. S1). Sea anemones Collection (ZRC), Lee Kong Chian Natural History
were observed and photographed in situ prior to Museum, National University of Singapore.
removing individuals (M. aspera: N = 12: M. fautinae
sp. nov.: N = 29) from the soft substratum using a hand- Molecular analyses
trowel. In the laboratory, tissue was sampled from the
animal’s pedal disc, column and tentacles, and preserved Genomic DNA from ethanol preserved tissues
in 100% molecular-grade ethanol for further molecular were isolated via standard CTAB extraction (see
analyses (see below). The animals were subsequently Rodríguez et al. 2014, and references therein). We
relaxed in 7.5% magnesium chloride and then fixed in utilised four published molecular markers in this study:
10% formalin. Identities of other sea anemone species, three mitochondrial (i.e., partial 12S rDNA: Chen et al.
for which their tissues were used for the molecular 2002; 16S rDNA and cox3: Geller and Walton 2001)
analyses in this study, were verified using evidence and one nuclear (i.e., partial 28S rDNA: Chen and
published in primary scientific literature (e.g., Fautin et Yu 2000). Loci for these markers were amplified with
al. 2009 2015; Fautin and Tan 2016). polymerase chain reaction (PCR), following published
cycling profiles (i.e., Lauretta et al. 2013; Rodríguez
Morphological observations et al. 2014 and references therein). All PCR products
were purified with SureClean Plus (Bioline, Singapore).
All fixed specimens were dissected so that the Thereafter cycle sequencing was performed using
internal morphology could be examined. Histological BigDye Terminator v3.1 (Applied Biosystems, Foster
sections, 8 µm thick, were prepared from these materials City, California) following the manufacturer’s protocol.
so that mesentery arrangements and musculature of the All amplicons obtained were sequenced on an ABI 3130
animal could be observed. All sections were stained XL Genetic Analyzer (Thermo Scientific).
with haematoxylin and counterstained with eosin New sequences acquired in this study were
(Humason 1967). assembled in Geneious v11.1.3 (Invitrogen Corporation)
Cnidae present in the formalin-fixed tissues of using default parameters. All contigs were searched
the sea anemone’s tentacle tip, marginal projections, against GenBank sequences via BLASTn; this was
mid-column, actinopharynx and mesenterial filaments, to affirm that nucleotide sequences we had obtained

and successfully amplified were only sea anemones. their similarity in appearance, M. aspera and D. armata
Selected sequences previously published were also were not recovered as a clade. The latter was recovered
used in our analyses (i.e., Rodríguez et al. 2014; Daly in a clade consisting of members of Actinodendridae
et al. 2017; Titus et al. 2019; Yap et al. 2021). Gene (i.e., A. arborerum and Ac. haeckeli, with strong support
sequences were aligned separately using MAFFT (i.e., bootstrap > 80) (Fig. 1).
v.7.313 under --auto setting. These aligned sequences Pairwise genetic distances between congenerics M.
were then concatenated into a combined matrix. In all, aspera and M. doreensis ranged from 2.4% to 16.1%;
separate analyses for alignments were conducted over dependent on the molecular marker examined (Table
the five resultant datasets (i.e., 12S, 16S, cox3, 28S and 1). Between M. aspera and M. fautinae sp. nov., these
concatenated matrix). genetic distances were smaller among the mitochondrial
Maximum likelihood (ML) analyses were markers (i.e., cox3, 12S and 16S: 0.8% to 2.0%).
conducted in RAxML-NG v0.8.1 (Stamatakis In contrast, genetic distances were larger in nuclear
2014; Kozlov et al. 2019) using 50 random and 50 markers (i.e., 28S: 6.4% to 10.1%; Table 1). While M.
parsimonious starting trees, with 1000 bootstrap pseudo- aspera may superficially resemble D. armata, pairwise
replicates. The best model of DNA evolution was differences of the mitochondrial markers between
determined using ModelTestNG (Darriba et al. 2020) these two species were larger than between members
and specified for all datasets in both ML analyses and of Macrodactyla, ranging between 3.0% and 3.8%
Bayesian inference (BI) described below. Uncorrected (Table 1). Dofleinia armata was recovered as a sister to
pairwise distances, for each alignment, was calculated the actinodendrids (Fig. 1), and this finding is further
using MEGA X (Kumar et al. 2018). supported here by the small pairwise genetic differences
Bayesian inference was carried out only on the (Table 1).
concatenated matrix using MrBayes v3.2.6 (Ronquist et
al. 2012). Four Markov Chain Monte Carlo (MCMC)
runs were carried out for 12 million generations and TAXONOMY
sampled at every 100th tree. To determine if the runs
converged, Tracer v1.7 (Rambaut et al. 2018) was used Order Actiniaria Hertwig, 1882
to visualise the posteriors. Convergence was achieved Suborder Enthemonae Rodríguez and Daly,
after discarding the first 60,001 trees as burn-in (average 2014 in Rodríguez et al. 2014
standard deviation of split frequencies = 0.0322, ESSLnL Superfamily Actinioidea Rafinesque, 1815
= 1152, ESSLnPr = 2860). Family Actiniidae Rafinesque, 1815
All new sequences obtained in this study are Genus Macrodactyla Haddon, 1898
deposited in GenBank (Table S1).
Condylactis [pro parte] – Haddon and Shackleton 1893: 123; Fautin
2016: 105.
Macrodactyla [pro parte] – Haddon 1898: 431 [original description];
RESULTS Stephenson 1921: 524, 531, 545; Stephenson 1922: 263, 265;
Carlgren 1949: 63; Dunn 1981: 28, 35; Fautin et al. 2007: 213;
Phylogeny reconstruction Fautin et al. 2008: 49; Fautin 2016: 104, 105.
Overall, 198 taxa were present in the concatenated Etymology gender: Feminine.
dataset, with 6,195 sites used for phylogeny Type species: Condylactis aspera Haddon and
reconstruction. Broadly, while trees inferred from Shackleton, 1893, by monotypy, in agreement with
both ML and BI analyses of the concatenated dataset Dunn (1981), Fautin et al. (2007) and Fautin (2016).
and four individual molecular markers were not fully Definition of genus modified after Dunn (1981)
concordant, topologies of phylogenetic relationships and Fautin et al. (2008). Substantial changes from our
inferred across all trees remained similar at genus level research are indicated in bold; all minor additions are
(Fig. 1; Figs. S3–S7). underlined.
In all inferred trees, Macrodactyla was found to be Diagnosis: Pedal disc distinct, circular to oval
polyphyletic, with M. aspera and M. doreensis failing to in outline, as wide as oral disc. Column elongated,
form a clade. Instead, M. doreensis was nested among marginal projections present. Adhesive verrucae
most members of Heteractis, with strong support. present on distal end, often with foreign materials
Macrodactyla aspera was recovered to be a sister to M. often attached, may be lobed and perforated. Fosse
fautinae sp. nov., forming a distinct clade (Fig. 1; Table very shallow. Tentacles long and stout, covered
1). This relationship is well-supported, with bootstrap with nematocyst batteries or entirely smooth;
values being > 50 at inclusive nodes (Fig. 1). Despite inner tentacles considerably longer than the outer.

Fig. 1. Phylogeny reconstruction of Macrodactyla aspera and Macrodactyla fautinae sp. nov. among actiniid sea anemones. Tree topology a result
of maximum likelihood (ML) analysis, using a concatenated dataset (i.e., cox3, 12S, 16S, 28S). At the branches, bootstrap ML resampling values,
and Bayesian inference (BI) posterior probability values, are presented as ML/BI. Only bootstrap values > 50, and posterior probability > 0.8 are
indicated here; those less than 50 and 0.8 are denoted by (-). Taxa indicated in bold denotes the presence of new sequences obtained for this study.
Indicated by an asterisk (*), we retain the name Macrodactyla doreensis in this figure to aid with the discussion within the main-text. For sequences
used and the full tree, refer to table S1 and figure S3, respectively.

Actinopharynx often everted when alive, with a pair Latreille (1825: 372) (Schoolmeesters 2021).
of diametric siphonoglyphs. Symmetrically-arrayed We posit that Harris’ entry in Hitchcock (1833)
mesenteries; always 12 pairs complete, all remainder was a misspelling of the coleopteran genus name
incomplete. All fertile. Retractor muscles band-like, Macrodactylus. Nomenclatural information we have
diffuse-circumscribed. Endodermal marginal sphincter found on Ma. subspinosus corroborated with much
muscle conspicuous and restricted. Parietobasilar of those present in Harris’ entry (i.e., taxonomic
muscles well-developed, with an extended pennon authority, species epithet, common name, etc.), as
present for lower order mesenteries. Cnidom: spirocysts, with this species’ geographical occurrence. Moreover,
basitrichs, microbasic p-mastigophores. Harris’ revised checklist in Hitchcock’s (1835: 565)
Nomenclatural considerations: Neave (1940) latter work strongly supports our assertion: in this
reports that the Macrodactyla, as a name for a sea updated listing, Harris attributed the taxonomic
anemone genus, is a junior homonym to a beetle genus authority of the misspelled beetle genus name (still
described by TW Harris in Hitchcock (1833). As listed as Macrodactyla) to PA Latreille. No works of
mentioned at the outset, Fautin (2016) had returned both Latreille prior to Harris’ entry in Hitchcock (1833) had
Macrodactyla species (i.e., M. aspera and M. doreensis) mentioned Macrodactyla, only the name Macrodactylus
to the next oldest available generic name applied to was ever used in Latreille’s (1825) publication.
them: Condylactis. The name Macrodactyla in Hitchcock (1833)
We consulted Harris’ contribution in Hitchcock is thus not a senior homonym to the genus name that
(1833: 575) and found the name Macrodactyla, Haddon (1898) had used for the sea anemones; Harris’
confirming its appearance in the publication. This listed entry is an incorrect spelling of the coleopteran genus
name is further accompanied by a species epithet, a name Macrodactylus, and therefore cannot enter
letter that denotes its taxonomic authority of the species homonymy (Article 54.3, the Code). As First Revisers,
(i.e., ‘F’ = JC Fabricius), and a common name – all of we retain the name Macrodactyla as a genus name for
which we render here exactly as (including the way it sea anemones; it is valid and available for use.
was stylised; see Hitchcock 1833: 575): (Macrodactyla) Ta x o n o m i c c o n s i d e r a t i o n s f o r t h e g e n u s
subspinosa, F. Rose-Bug. Macrodactyla: Historically, sea anemone systematists
Harris’ entry in Hitchcock (1833) was meant have considered Haddon’s (1898) report of mesentery
simply to be part of an inventory of the insect biota number and its arrangement in Macrodactyla to be
of Massachusetts, and it lacks an accompanying ambiguous. Based only on the type species, C. aspera,
description to Macrodactyla. Haddon defined Macrodactyla to have “six pairs of
No beetle species is known by the name listed by imperfect [= incomplete] mesenteries, and a second
Harris in Hitchcock (1833). The closest match is the and third cycle of imperfect mesenteries” (Haddon
species Macrodactylus subspinosus (Fabricius, 1775), 1898: 431; additions in square parentheses, our
which also occurs in Massachusetts, and is known by own). Stephenson (1921) later revised this feature to
the common names, ‘Rose-bug’ and ‘Rose-chafer’ (see: denote the presence of only six complete mesenteries,
Blatchely 1910: 953; Schoolmeesters 2021). Originally elaborating further that Haddon (1898) may been
described by Fabricius (1775) as a member of the mistaken on this feature (Stephenson 1922: 263). Later
genus Melolontha Fabricius, 1775, this species was definitions of Macrodactyla retained Stephenson’s
later placed in a new genus, Macrodactylus, created by assertions (e.g., Carlgren 1949). By including a second
Table 1. Pairwise genetic distances among Macrodactyla, Dofleinia, and members of family Actinodendridae (i.e.,
Actinodendron and Actinostephanus) for the four molecular markers targeted in this study. Dashes indicate that no
comparison were made, as we failed to obtained 28S sequences from D. armata (see Table S1)
Pairwise genetic distance range
M. aspera M. aspera M. aspera D. armata

Molecular markers and and and and
M. doreensis M. fautinae sp. nov. D. armata actinodenriids
Mitochondrial cox3 5.3–6.5% 1.7–2.0% 3.0–3.6% 1.0–1.5%

12S 2.4–2.8% 0.8–0.9% 2.5–2.8% 0.3–3.01%
16S 2.4–3.0% 0.9–1.09% 3.3–3.8% 1.3–1.5%
Nuclear 28S 10.8–16.1% 6.4–10.1% - -

species to the genus, M. doreensis, Dunn (1981: 28) had M. aspera are always adhesive, whereas verrucae of
amended its definition to include, “… six or more pairs M. doreensis are not (Dunn 1981; Fautin et al. 2008).
of mesenteries perfect,… ” because this second species Mesentery number and arrangement between them also
had more than six complete pairs of mesenteries. differ; those in M. aspera are arranged in up to three
However, in Dunn’s [as Fautin] later publication of cycles while those of M. doreensis may go up to seven
this genus, its diagnosis was changed, indicating that (Dunn 1981). Furthermore, we found microcnemes to
only six pairs of complete mesenteries were present in be present in M. doreensis, consistent to its depiction in
Macrodactyla (see Fautin et al. 2008: 49). In studying Dunn (1981: Fig. 13), a trait that is absent in M. aspera.
the type specimen of the type species for Macrodactyla Ecologically, M. doreensis is a host to photosynthetic
(i.e., C. aspera; MZC.I.33665), we found that it has 12 dinoflagellates and macro-fauna such as clownfishes
pairs of complete mesenteries instead of six, a feature (Dunn 1981; Fautin and Allen 1992). In contrast, we
that is consistent across all voucher and fresh material. have not encountered nor observed M. aspera to be
Here, we have revised the definition of the genus to involved in any symbiotic relationships with other
reflect this. micro/macro-organisms.
No data concerning the cnidom for the type Given the genetic, morphological and ecological
species of Macrodactyla (i.e., C. aspera) have ever evidence presented, we place M. doreensis in the
been reported, it being absent in earlier descriptions of genus Heteractis, under a new combination: Heteractis
the genus (i.e., Haddon and Shackleton 1893; Haddon doreensis (Quoy and Gaimard, 1833). In revising the
1898; Stephenson 1921 1922). In recent definitions diagnosis of Macrodactyla, we removed all characters
of Macrodactyla, which had included M. doreensis, that had extended to M. doreensis (e.g., presence of
Fautin et al. (2008: 49) simply reported the types of deep fosse).
cnidae present in sea anemones of the genus, stating
the presence of, “spirocysts, basitrichs, microbasic Macrodactyla aspera (Haddon and Shackleton,
p-mastigophores”. We confirmed that these cnidae were 1893)
present in the name-bearing type specimen of C. aspera. (Figs. 2–6)
We deemed Macrodactyla to be the most
appropriate genus to place the new species, M. fautinae Condylactis aspera – Haddon and Shackleton, 1893: 117, 124–125
[original description]; Dunn 1981: 29; Fautin 2016: 77, 105, 164.
sp. nov., in the light of both morphological, ecological Macrodactyla aspera – Haddon 1898: 398, 415, 431–432; Stephenson
and genetic evidence gathered from this study. Both M. 1922: 263; Carlgren 1949: 63; Fishelson 1970: 109, 110;
fautinae sp. nov. and M. aspera share the same number Stephenson 1922: 263; Dunn 1981: 35.
and arrangement of complete mesenteries. Both species Dofleinia armata – Rowlett 2020: 292, 293.
also have visible adhesive verrucae on their column,
and harbour the same types of cnidae in their tissues Nomenclature considerations and type material
(see below, in their respective species description). examined: Two lots of the syntypes of C. aspera that
Like M. aspera, we did not find any symbiotic micro/ was listed in Fautin (2016), were of sea anemones
macro-organisms associated with M. fautinae sp. nov. collected from Murray Islands in the Torres Strait,
Our placement of this new species in Macrodactyla is Australia: i) a nearly complete specimen kept at the
further supported by genetic evidence we have gathered: Cambridge, UK (MZC I.33665, previously without
we had repeatedly recovered both M. fautinae sp. nov. a catalogue number assigned) (Fig. 2A), and ii) three
and M. aspera to be sister to each other, within a clade, microscope slides comprising of histological sections
with strong support (bootstrap > 80; Fig. 1). Our revised kept in Lund, Sweden (MZL, no catalogue number)
diagnosis of Macrodactyla now includes traits of this (Fig. 2B). We confirmed Fautin’s (2016) listing of these
second species, thereby expanding on its definition. specimens; the labels present on these slides indicated
We failed to recover M. doreensis and M. aspera that they were prepared from the material collected by
as a monophyletic clade (Fig. 1). Genetic evidence from Haddon and Shackleton (1893) (Fig. 2B). Among these
this study, as with observations from recent studies, three microscope slides, only histological sections of
have repeatedly recovered M. doreensis to be nested one closely corresponded to Haddon’s depiction of the
among clownfish and dinoflagellate-associated sea musculature, while those present on the other two slides
anemones of the genus Heteractis (Titus et al. 2019; did not agree to his remaining illustrations (see: Haddon
Yap et al. 2021; Fig. 1). Apart from genetic evidence, 1898: Plate 28, Figs. 3, 4 and 5). Despite this, sections
both M. aspera and M. doreensis also do not share on these slides agreed with the shape of pieces that
many morphological and ecological traits that would were missing from the syntype which we had studied
characterise them to be classified in the same genus. at the MZC. These other histological sections from the
While both have verrucae on the distal end, those of syntype were likely prepared by O Carlgren, who then

kept them at the MZL where he had worked, a practice original description for the species, the account of the
which he also carried out on other sea anemone species species’ tentacle arrangement alludes to more than one
(Fautin 2016: 13; Yap et al. 2021). Pertaining to the individual being examined, as Haddon and Shackleton
original histological sections that were prepared by (1893: 124) stated, “… large tentacles in three or
Haddon and used for his illustrations, we were unable to four cycles…” [underlined emphasis, our own]. This
locate it at the museums we have visited. observation of a variation is only possible if more than
To further provide nomenclatural objectivity to one individual was examined. In this study, NWL Yap
the name Condylactis aspera, we hereby designate attempted to locate other syntypes in other museums
MZC.I.3365 and the three MZL slides as the lectotype, where they may be possibly held (e.g., NHM, etc), but
as the latter slides were prepared from the specimen failed to find any. Should these other materials be found,
at MZC. While there was no mention of the number they are the paralectotypes of C. aspera (Article 74.1.3,
of individuals that were collected and examined in the the Code).
Fig. 2. Type material of Condylactis aspera Haddon and Shackleton, 1893. A, lectotype of C. aspera (MZC I.33665), in three pieces, top view.
Note the gastro-cavity of specimen is filled densely with gametogenic tissue. B, Paralectotypes of C. aspera, as three histological slides at MZL (no
catalogue number). Abbreviations: gt, gametogenic tissue; m, mesenteries; o, oral disc; s, siphonoglyph; t, tentacle. Scale bar = 10 mm.

Lectotype: Condylactis aspera (MZC I.3365), Tentacles: 48. Arranged in four cycles, one per
collected by Haddon and Shackleton from the Torres endo-/exocoel. Outermost exocoelic; innermost cycles:
Straits (Fig. 2A). A single specimen cut into three endocoelic. Innermost cycle longer, greater or equal
pieces. Two pieces already removed prior to being to length of oral disc radius; outermost shortest, 1/3
examined by the first author: i) a wedge of the oral disc, length of innermost (Fig. 3A). Covered entirely with
and ii) a quarter section of the base. Column diameter visible nematocyst batteries (Figs. 3A–D, 4A, 4B).
about 5 mm. Entire specimen greyish, with a dark Tapering in life; may appear conical in fixed specimens
brown cast (Fig. 2A). Gastro-cavity of specimen choked due to contraction by fixative (Fig. 4B). Wide at base,
with gametogenic tissue. narrows to point. Tip pointed-blunt, not perforated. In
Lectotype: MZL (no catalogue number), three life, cream-colour with dark-brown splotches on oral
microscope slides prepared from MZC.I.3365 (Fig. 2B). side, batteries white or brown (Figs. 3A–D, 4A). Sticky;
One labelled as “Macrodactyla aspera Torres Str AMS readily adheres to surfaces of containers. Does not
1893, consists of three transverse sections of the distal abscise readily when disturbed. Preserved specimens
most end of the column and tentacle. Second slide, coloured grey, brown to mauve, with intense brown (or
with two labels. The first wider label bears the words, dark mauve) cast near the tip (Fig. 4B).
“Macrodactyla aspera sph”, the second slimmer label Oral disc: Outline round, flat when fully expanded
has “AMS 187” written on it. The slide bears eight (Fig. 3A). Diameter up to 45 mm. Nematocyst batteries
sections, all being the distal most end of the specimen. of tentacles may extend across disc, in some specimens
The third slide has a single label with the following all the way to mouth. Thin-walled; mesenterial
printed on it: “Macrodactyla aspera Torres Str, AMS insertions as radial lines seen through disc, extending
1893”. This slide also bears three histological sections from mouth to tentacles, in life these appear as faint
of the distal most end of the animal. All sections present brown lines (Fig. 3A, B), in preserved specimens as
on these slides are too faded to infer any useful details white lines. Oval central-mouth, flat, may be slightly
from them. raised in some. Actinopharynx often everted (Fig.
Material examined from Singapore (Fig. S1): (*- 3B), sometimes obscuring entire animal. When alive,
observed alive): Beting Bronok (ZRC.CNI.1015 x3), edge of mouth typically white in colour, with thick
Changi beach (ZRC.CNI.1055 x1; ZRC.CNI.1099 brown splotches radiating outwards, translucent when
x1*; ZRC.CNI.1217 x2*), Changi East beach (ZRC. expanded. In preserved materials, edge of mouth with a
CNI.1090 x1*), Cyrene Reef (ZRC.CNI.1080 x1; brown cast.
Photographed but not collected), Pulau Sekudu (ZRC. Column: Thin-walled. Fosse present, very shallow
CNI.1222 x3*). in vivo; in preserved specimens fosse not distinct. When
Materials examined from elsewhere (Fig. S2): expanded, distal end flared outwards, overhangs slightly.
Israel (Eilat): Gulf of Aqaba (SMNHTAU-Co.7813 x1; Distal end with marginal projections, endocoelic (Fig.
SMNHTAU-Co.12133 x1). Australia (Perth): North 3C, D). Longitudinal rows of conspicuous verrucae
Fremantle (WAM Z99100 x1), Cockburn Sound (WAM present, extending from distal end to mid-column, 6
Z50023 x1); Rowley Shoals (WAM Z50026 x2). New to 13 per row; endocoelic (Fig. 3C, D). Verrucae oval,
Caledonia: Noumea (MNHN 1321 x1); Unspecific edges slightly raised and thickened, middle thin and
location (MNHN 1571 x1). translucent; in life, verrucae cream-white with a pink
Description: Natural history: Usually solitary, at dot in the middle (Fig. 4C), colour is lost in preserved
the middle intertidal of soft silty sand, sandy areas, and specimens (Fig. 4D). Verrucae adhesive, with small
among seagrasses (Fig. 3A–B). Distal end of animal shell fragments, sand, small stones, and/or coral rubble
exposed during low spring tides, when encountered often attached (Fig. 3C, D) even after preservation (Fig.
during the early morning or dusk. Animal partially 4D). Mesenterial insertions seen as light lines extending
buried in substratum (Fig. 3A–B). Tentacles extended from the distal to proximal end. Remainder of column
and oral disc expanded, flat against substratum (Fig. smooth, no other structures present (e.g., cinclides).
3A). Often encountered with actinopharynx everted In life, distal end brown, region from mid-column to
outwards; if everted partially, animal may appear to proximal end bright orange; once preserved, specimen
have luscious lips (Fig. 3B). Small shell pieces, coral may be entirely cream-white, dark brown or grey.
rubble, sand and/or small stones adhere to exposed Pedal disc: Overall oval in outline, flat; thin-
verrucae of distal most column end (Fig. 3C–D). When walled. Mesenterial insertions seen through as white
disturbed, animal does not retract completely, with lines. Limbus slightly scalloped. In life, orange with
tentacles and/or its tips remaining partially exposed (Fig. splotches; once preserved entirely cream-coloured or
3D). Cells of symbiotic dinoflagellates not observed in greyish, with a dark-brown cast.
tissue. Internal morphology: 24 pairs of mesenteries

regularly arrayed in three cycles (6 + 6 + 12), with two difficult to view (e.g., SMNHTAU-Co.12133).
pairs of directives symmetrically opposite each other, Parietobasilar muscle present but not easily discernible.
each pair attached to a siphonoglyph. Mesenteries Actinopharynx extends proximally slightly past mid-
in primary and secondary cycles complete (i.e., 6 column, longitudinally pleated, pinkish translucent in
+ 6 pairs); those in third cycle, incomplete (i.e., 12 live, cream-white when preserved (Fig. 5C).
pairs). All mesenteries extended through entire length Cnidom: Spirocysts, basitrichs, and microbasic
of column. Mesenterial retractor muscles diffuse- p-mastigophores. Sizes and distribution are shown in
circumscribed, well-developed (Fig. 5A). Parietobasilar table 2; illustrations in figure 6.
muscles of first two cycles well-developed, with pennon Distribution (Fig. S2): Type locality: Australia,
(Fig. 5A). Marginal sphincter muscle conspicuous, Torres Straits, Mabuiag Reef (Haddon and Shackleton
diffuse-circumscribed. Oral and marginal stomata 1893; Haddon 1898). Published records: Israel, Red
present (Fig. 5B), but in some specimens may be Sea, Gulf of Aqaba, Eilat (Fishelson 1970). New
Fig. 3. Living individuals of Macrodactyla aspera, external morphology. A, B, expanded individuals extending from the substratum, in situ, top
view. Note splotchy brown patterns on oral disc and tentacles in both A and B, and partially everted actinopharynx resembling thick lips in B.
Photographs by KS Loh. C, a live collected specimen (ZRC.CNI.1099), side view. Note reddish-orange column and adhesive, conspicuous verrucae,
with grains of substratum attached to it. D, a partially retracted specimen (ZRC.CNI.1099), top view. Note that individual does not completely retract
its tentacles; note also the presence of marginal projections. Abbreviations: mp, marginal projections; v, verrucae. Scale bars = 10 mm.

records: Australia, Indian Ocean, Perth (i.e., Cockburn adhesive verrucae) hidden in the sand, and in sharing
Sound, North Fremantle, and Rowley Shoals); New a similar habitat, M. aspera have been misidentified
Caledonia, Coral Sea, Noumea; Singapore, in the Straits as D. armata (e.g., WAM Z99100), or assumed to be a
of Johor (i.e., Beting Bronok, Changi region beaches, different morphotype of the latter (i.e., Rowlett 2020:
Pulau Sekudu) and Singapore Strait (i.e., Cyrene Reef). 292, 293). However, the presence of longitudinal,
Comparisons with other similar sea anemones: adhesive verrucae rows is the obvious feature that
In having nematocyst batteries densely covering its distinguishes these two species. These rows are only
tentacles, other discerning taxonomic features (e.g., present on the distal end of M. aspera, while they
Fig. 4. Close-up appearance of tentacles and distal most column of Macrodactyla aspera, external view. A, extended tentacles of a live specimen (ZRC.
CNI.1080), covered densely with nematocyst batteries. B, tentacles of a fixed specimen (i.e., lectotype, MZC.I.33665). Note the overall stout, conical
shape of tentacles once fixed, and the bumpy texture for each of them, indicating the presence of nematocyst batteries. C, marginal projections and
adhesive verrucae of a live individual (ZRC.CNI.1080). D, marginal projections and adhesive verrucae of a fixed specimen (SMNHTAU-Co.7813).
Note how the shallow fosse had inflated. Also note how the grains of substratum remained attached to the specimen, when it was collected by L
Fishelson in 1968. Abbreviations: f, fosse; mp, marginal projections; nb, nematocyst batteries; v, verrucae. Scale bars = 10 mm.

are absent in D. armata; the absence of verrucae is

consistent with taxonomic accounts of D. armata and
its holotype (Wassilieff 1908; Carlgren 1949). When
alive, another trait that distinguishes D. armata from
M. aspera is that the former readily autotomises its
tentacles when disturbed, but those of the latter will
remain intact.
Remarks: The animal that we have found closely
matches the species and its type specimen that was
collected and described by Haddon and Shackleton
(1893). In all these specimens studied, we found that:
i) their tentacles were densely covered with nematocyst
batteries, ii) visible, adhesive verrucae were present
at the distal end of the column, iii) cnidom data
agreed between these specimens, and iv) as stated
in our revised definition of the genus, the mesentery
arrangement and number were also consistent.
Although the distinct tentacles of M. aspera (i.e.,
it being densely covered by nematocyst batteries) were
not mentioned in its Haddon and Shackleton’s (1893)
original description of the species, nor in the main
text of Haddon’s (1898) expanded account, they were
illustrated in the latter (see: Haddon 1898: Plate 22,
Figs. 10 and 11). His depiction also provided a close-
up view of a studded tentacle, clearly showing the
nematocyst batteries. In examining the lectotype, Dunn
(1981: 35) also found this feature to be obvious. The
presence of nematocyst batteries densely covering each
tentacle was also observed in fresh material collected,
and in museum voucher specimens we have found and
studied. Because the tentacles of M. aspera strongly
resemble those of D. armata, many museum vouchers
of the former species were identified and labelled as
the latter (e.g., MNHN 1321, WAM 67). We have also
examined specimens that Fishelson (1970) had collected
(i.e., SMNHTAU-Co.7813 and SMNHTAU-Co.12133),
and verified them to be M. aspera.
Like Haddon and Shackleton (1893), we found
large visible, adhesive verrucae present on the distal end
of the anemone, with shell fragments and small rocks
often attached to them. While Haddon and Shackleton
(1893: 124) also mentioned that the column of the
anemone was also “covered with small, very adhesive
Fig. 5. Macrodactyla aspera, internal morphology. A, mesenteries
suckers,” we failed to find any structural indication
of the lectotype (MZC.I.3365), cross section at mid-column. Note of this feature in both the lectotype and the voucher
the diffuse circumscribed appearance of the retractor muscles, and specimens.
presence of the retractor pennon. B, transverse section of the distal The few published accounts pertaining to
most end of column (SMNHTAU-Co.7813). Note the presence of a this species and its lectotype provided no detailed
conspicuous, restricted marginal sphincter muscle and the presence measurements of cnidom data (e.g., Haddon 1898;
of both oral and marginal stomata. C, everted actinopharynx of a
Fishelson 1970; Dunn 1981). We report these here
live specimen (ZRC.CNI.1090). Note its pinkish appearance, and
the presence of a diametric pair of siphonoglyphs. Abbreviations: a,
for the first time. Broadly, measurements made on
actinopharynx; mf, mesenterial filaments; ms, marginal stomata; o, undischarged capsules extracted from tissues of the
oocytes; os, oral stomata; p, pennon; s, siphonoglyph; sph, marginal voucher material fell within the range of those obtained
sphincter muscle. Scale bars = 5 mm. from the lectotype (Table 2). We were unable to assess

Fig. 6. Cnidae of Macrodactyla aspera. A, spirocyst. B, small basitrich. C, basitrich. D, large basitrich. E, small basitrich. F, basitrich. G, small
basitrich. H, basitrich. I, small basitrich. J, large basitrich. K, wide basitrich. L, small basitrich. M, large basitrich. N, wide basitrich. O, microbasic
p-mastigophore. Scale bar = 10 µm. Refer to table 2 for key to letters.
Table 2. Cnidom of Macrodactyla aspera (Haddon and Shackleton, 1893). Sizes of single capsules that fell out of the
size range for the cnida type measured are indicated as numerical values in parentheses
Lectotype MZC.I.33665 Vouchers
Tissue Cnidae Range length × width (µm) n Range length × width (µm) n N
Tentacles Spirocysts (A) - - (17.0)20.0–30.3 × 1.5–2.5 40 4/4

Small basitrichs (B) 5.0–11.0 × 2.0–3.0 10 8.0–12.0(15.0) × (1.5)2.0–3.0 30 3/4
Basitrichs (C) 30.0–36.4 × 2.5–3.5 18 17.0–32.0(34.0) × 2.0–3.5 40 4/4
Large basitrichs (D) 37.9–54.5(57.5) × (2.0)2.5–3.5 20 33.0–50.0 × 2.5–3.5 40 4/4
Marginal projections Small basitrichs (E) 9.0–12.0(15.0) × 2.0–3.0 10 7.0–15.0 × 2.0–3.0(4.0) 40 4/4
Basitrichs (F) 17.0–20.5 × 3.0–3.5 10 (17.7)18.0–22.2 × 2.5–3.0 40 4/4
Column Small basitrichs (G) 9.0–11.0 × 2.0–3.0 19 6.0–12.0(15.7) × 2.0–3.5 40 4/4
Basitrichs (H) (15.0)18.0–22.0 × (2.0)2.5–3.0 20 (16.0)17.0–24.2 × 2.0–3.0 50 4/4
Actinopharynx Small basitrichs (I) - - (7.0)9.0–17.0 × (1.5)2.0–3.5 40 4/4
Large basitrichs (J) - - 30.5–42.0 × 4.0–6.5 40 4/4
Wide basitrichs (K) - - 34.0–41.0(43.0) × 7.0–14.0(18.0) 40 4/4
Mesenterial filaments Small basitrichs (L) 9.5–14.0 × 2.0–3.0 10 7.0–15.5 × 2.0–3.0 40 4/4
Large basitrichs (M) (31.0)35.0–43.5 (46.0) × 3.0–4.0 10 32.0–47.0 × 3.0–5.0 40 4/4
Wide basitrichs (N) 50.0–59.0 × (4.0)5.0–6.0(7.0) 10 (32.0)35.0–55.0 × 6.0–12.0 40 4/4
Microbasic p-mastigophores (O) (23.0)26.0–29.5 × (4.5)5.0–6.0 10 25.0–35.0 × 4.0–5.5(6.0) 40 4/4
Abbreviations: N, the number of specimens for which the type of cnida was present to the total number of specimens examined; n, the number of
cnida capsules measured in this study. Refer to figure 6 for illustrations corresponding to the letters in parentheses.

cnidae of the lectotype’s actinopharynx as we failed to (near Carpark 7), Singapore, on 5 July 2016 by the staff
obtain a piece of tissue for it. Pertaining to the tentacle of LKCNHM. An individual of unknown gender, cut
spirocysts of the lectotype, we found these to be longitudinally into two equal pieces. Tissue from the
degraded in the tissue, hindering measurements. Despite pedal disc sampled for genetic work (Table S1).
these two caveats, cnidae measurements of the lectotype Paratypes: ZRC.CNI.1156, collected from Changi
and voucher materials agreed well. Beach (near Carpark 4) (Fig. S1), Singapore, on 4
August 2016 by NWL Yap. Tissue sampled from its
Macrodactyla fautinae, sp. nov. Yap, Mitchell, pedal disc for genetic work (Table S1). ZRC.CNI.0296,
Quek, and Huang collected from Changi Point Beach on 22 July 2009
(Figs. 7–11) by DG Fautin et al. A dissected female from which
urn:lsid:zoobank.org:act:DC99CC5F-0956-47A9-B0E9- 10 microscope slides with histological sections of the
A0B3206F5ADE animal were prepared.
Material examined from Singapore (Fig. S1)
Material examined: Holotype: ZRC.CNI.1300 (*- observed alive): Changi Beach (ZRC.CNI.1020
(Fig. 7), collected from the beach off Changi Beach
Fig. 7. Holotype of Macrodactyla fautinae sp. nov. (ZRC.CNI.1300). Abbreviations: a, actinopharynx; m, mesenterial filaments; mp, marginal
projections; pd, pedal disc; s, siphonoglyph; sph, sphincter muscle; t, tentacles; v, verrucae.

x5*; ZRC.CNI.1156 x1*; ZRC.CNI.1159 x1*; ZRC. typically larger individuals; see also Fig. 8A, B); disc in
CNI.1300 x1*), Changi Point beaches (ZRC.CNI.0022 fixative all appear translucent cream-coloured. Central
x1; ZRC.CNI.0025 x1; ZRC.CNI.00296 x1; ZRC. mouth lipless and oval in outline, with actinopharynx
CNI.00297 x1), Changi Point SAF Chalet beach (ZRC. everted to some extent (Fig. 8C, D). Two symmetric
CNI.0558 x1*; ZRC.CNI.0559 x1*; ZRC.CNI.0566 siphonoglyphs, each marked by bright pink dot in live
x1*; ZRC.CNI.0598 x1*; ZRC.CNI.0648 x3*), Changi individuals (Fig. 8A, B); pink dot may be visible in
Point Ferry Terminal beach (ZRC.CNI.0023 x1; ZRC. freshly preserved specimens but fades over time.
CNI.0024 x1), Chek Jawa (ZRC.CNI.1370 x1*), Pasir Column: In life, translucent cream-coloured, with
Ris (ZRC.CNI.00254 x1; ZRC.CNI.00255 x1), Pulau reddish-orange splotches present on lower end (Fig.
Ubin, OBS Camp beach (ZRC.CNI.0788 x1*), Pulau 9A); once fixed, colour may be retained in specimens,
Sekudu (ZRC.CNI.0026 x1). but fades over time. Fosse present, shallow to deep.
Etymology: The name Macrodactyla fautinae sp. Thin-walled, mesenterial insertion visible through wall
nov. honours the late Emeritus Professor Daphne Gail as longitudinal light lines, extending from distal most to
Fautin. Throughout her career, she had worked tirelessly proximal end (Fig. 9B); visibility most prominent when
to advance the knowledge of sea anemones. animal is expanded and/or relaxed, may be obscured
Description: Natural history: Occurs as solitary when animal is slightly contracted (Fig. 9A). Distal
individuals, at the middle intertidal in soft silty, sandy end with marginal projections, perforated, both endo-/
areas, and also found in seagrass meadows (Fig. 8A– exocoelic (Fig. 9C). Conspicuous adhesive, verrucae
E). Animal partially buried in substratum, distal end of present, endocoelic, arranged in vertical rows that
animal exposed during the low tides, with small shell extend proximally from the distal end to mid-column,
pieces, sand and/or rocks adhering to verrucae (Fig. five to seven per row. Verrucae raised, outline round,
8C). Oral disc and tentacles typically expanded, with with a pink dot in the middle (Fig. 9B, C), perforated.
actinopharynx often everted and inflated outwards, in When extended, verrucae papillae-like (Fig. 9B). In
many instances obscuring the animal (Fig. 8D). This fixed specimens, verrucae contracted but still prominent.
species has also been observed to swallow its prey From mid-column to proximal end, without any obvious
whole (Fig. 8E). When disturbed, animal does not structures (e.g., cinclides, etc.); smooth.
retract completely, tips of tentacles remaining partially Pedal disc: Outline irregular oval in outline,
exposed (Fig. 8C). In the process of contraction, water following contours of surface it clings onto. Thin-
may also be expelled from verrucae-like structures, akin walled; mesenterial insertions visible through it as
to a watering can. radiating while lines. Scattered reddish-orange splotches
Tentacles: 96. Arranged in five cycles, one per present on base. Adheres readily to surfaces.
endo-/exocoel. Outermost exocoelic; innermost cycles, Internal morphology: 48 pairs of regularly
endocoelic. Innermost cycles longer and of similar arrayed mesenteries arranged in four cycles (6 + 6 +
length, greater or equal to length of oral disc radius; 12 + 24); all extend the entire length of column. Two
outermost shortest, ½ length of innermost cycles (Fig. pairs of diametric directives, each pair attached to a
8A, B). All alike in appearance; simple, smooth without siphonoglyph. Mesenteries in first two cycles complete,
nematocyst batteries, tapering to a blunt end in life; in those in third and fourth incomplete. Gametogenic
preservative all appeared similarly sized, short, conical tissues on all mesenteries, if present. Sexes separate.
with a blunt end, possibly due to contraction in fixative. Mesenterial retractor muscles well-developed, diffuse-
Colour in life translucent with horizontal bands of dark circumscript (Fig. 10A); mesenteries of lower orders
brown and white bands (Fig. 8A, B); in fixative, colours more circumscribed than those higher. Parietobasilar
retained in freshly preserved specimens (i.e., < two well-developed; free pennon associated with mesenteries
years) but are expected to be lost over time, to become of lower orders (Fig. 10A). Oral and marginal stomata
translucent cream in colour. present, however former may be difficult to visualise
Oral disc: Outline round, flat when expanded (Fig. in some individuals (e.g., ZRC.CNI.0566). Marginal
8A, B). Diameter up to 80 mm. Smooth and thin-walled, sphincter muscle conspicuous and circumscribed, close
mesenterial insertions may be seen as dark radial lines to fosse (Fig. 10B). Parietobasilar muscle present but
extending from mouth to disc margin, more prominent not easily discernible. Actinopharynx longitudinally
in individuals with colourless disc or in fixed specimens pleated and thin-walled, translucent-white in life, cream-
(e.g., Fig. 8A). In life, colour and pattern variable coloured in preserved specimens; extends proximally
among individuals: may be completely colourless, towards pedal end, slightly pass mid-column.
greenish-grey, entirely dark-brown, or brown with white Cnidom: Spirocysts, basitrichs, and microbasic
markings or vice-versa, sometimes accompanied by p-mastigophores. Sizes and distribution are shown in
markings of cross bands being V- or W-shaped (seen table 3; illustrations in figure 11.

Fig. 8. Living individuals of Macrodactyla fautinae sp. nov., external morphology, in situ, top view. A, an expanded individual with a pale oral disc. B,
another expanded colour morph with a dark-brown oral disc. Note that in both A and B, a pair of diametric bright pink dots marks the position of the
siphonoglyphs. C, a contracted individual with adhesive, papillae-like verrucae at its distal end. Note that shell fragments are attached to the verrucae.
D, an individual with much of its actinopharynx everted, obscuring the animal. E, a sea pen (Pteroeides sp.) being swallowed whole by a M. fautinae
sp. nov. individual. Abbreviation: s, siphonoglyph; v, verrucae. Photographs by R Tan.

Distribution: This species has so far been often marked by dark brown splotches, whereas those
encountered only along the northern shoreline of of M. fautinae sp. nov. may vary in appearance, and
Singapore, in the Straits of Johor (Fig. S1). markings – if present – are typically cross bands that are
Comparisons with M. aspera: The most obvious V- or W-shaped. Consistently in all individuals we have
feature that distinguishes these two species is the encountered and collected, bright pink dots demarcate
texture of the tentacles of their type species. Those of the siphonoglyphs of M. fautinae sp. nov., a feature not
M. aspera appear rough to the eye, as they are densely observed on the oral disc of live M. aspera individuals
covered with nematocyst batteries, while the tentacles examined (see Figs. 3A, B; 8A, B).
of M. fautinae sp. nov. are smooth. We have not The appearance of the column also differed
encountered any individuals of M. fautinae sp. nov. that consistently between the two species. In general,
bear nematocyst batteries on their tentacles. colour may not be a useful feature when distinguishing
Oral disc appearance also differed between the between sea anemone species (Stephenson 1928:
two type species. As with the tentacles, nematocyst 69), however, we found this feature to be consistent
batteries are often on the oral disc of M. aspera, while as a whole in all individuals of both species studied.
M. fautinae sp. nov. lacks this feature. In many of the Macrodactyla aspera has a dark-coloured distal-end
examined live M. aspera individuals, the oral disc is and the remainder of its column being bright orange,
Fig. 9. Column appearance of Macrodactyla fautinae sp. nov., external morphology, side view. A, a freshly collected individual that is slightly
contracted (ZRC.CNI.0648). Note the pale pinkish column and the presence of splotchy dark pink patches from mid-column to the proximal end.
Note also that the mesenterial insertions are not visible when the anemone is in a contracted stated. B, a live individual (ZRC.CNI.1159) with a
column that is expanded, with mesenterial insertions that extend the entire column visible as light lines. Note also the extended papillae-like verrucae.
C, close-up of the marginal projections and verrucae at the distal end of a live individual (ZRC.CNI.1159). Note that marginal projections are
perforated, and a pink dot marks the middle of each verruca. Abbreviations: mi, mesenterial insertions; mp, marginal projections; v, verrucae. Scale
bars = 10 mm.

whereas that of M. fautinae sp. nov. is entirely cream-

white, with pink splotches. Columnar structures also
differ between both species. While verrucae found in
these two species are adhesive, those of M. fautinae
sp. nov. are lobe-like when extended, and expels jets of
water when the animal contracts, a feature not observed
in M. aspera.
Ecological remarks: Both M. fautinae sp. nov. and
M. aspera are found to occur in similar habitats along
Singapore’s northern coastline, in the Straits of Johor.
However, while we have found the latter to also occur
in intertidal areas of Singapore’s southern shoreline
(i.e., in the Singapore Strait), we have only encountered
M. fautinae sp. nov. on the northern shoreline and in
no other southern locations that bear a similar habitat
(Fig. S1). Apart from sharing similar habitat types, both
species also exhibit a curious behaviour of everting
an extensive part of the actinopharynx when exposed
during the low tide, obscuring most of the animal. The
exact rationale of this behaviour is unclear. Inferring
from the presence of nematocysts present in the tissue
of the actinopharynx and the extent of the eversion, we
hypothesize that this behaviour aids in the capture of
larger prey, as we have observed (Fig. 8E). To test this
hypothesis, further experimental studies involving prey
size and measuring the extent of actinopharynx eversion
may be conducted.
Similar species: Macrodactyla fautinae sp. nov.
may be confused with three other similar looking
Fig. 10. Macrodactyla fautinae sp. nov., internal morphology. A,
diffuse-circumscript mesenteries of an individual (ZRC.CNI.0296),
species that also occur at its type locality, Singapore:
cross section at mid-column. Note the presence of the retractor Anthopleura buddemeieri Fautin, 2005, An. handi Dunn,
pennon. B, a conspicuous, circumscribed marginal sphincter muscle 1978, and Paracondylactis singaporensis (England
of another individual (ZRC.CNI.0255). Abbreviations: f, fosse; mf, 1987).
mesenterial filaments; p, pennon; sph, marginal sphincter muscle; t, While both M. fautinae sp. nov. and An.
tentacles. Scale bars = 1 mm. buddemeieri have a column patterned with splotches
Table 3. Cnidom of Macrodactyla fautinae sp. nov. Sizes of single capsules that fell out of the size range for the cnida
type measured are denoted as numerical values in parentheses
Tissue Cnidae Range length × width (µm) n N
Tentacles Spirocysts (A) 15.0–27.5 × 2.0–3.0 53 5/5

Large spirocysts (B) 30.0–41.0(43.0) × 2.5–5.0 47 5/5
Basitrichs (C) (9.0)10.0–20.0 × 2.0–3.0 51 5/5
Large basitrichs (D) 27.5–33.0(34.0) × 2.0–3.0 50 5/5
Marginal projections Basitrichs (E) 17.0–21.0 × 2.0–3.0(3.5) 50 5/5
Column Basitrichs (F) (10.5)15.0–20.0 × 2.0–3.0 52 5/5
Actinopharynx Basitrichs (G) (23.0)24.0–31.0 × 2.5–4.0 50 5/5
Microbasic p-mastigophores (H) 15.0–23.0 × 4.0–6.5(7.0) 52 5/5
Mesenterial filaments Small basitrichs (I) 10.0–13.0(14.0) × 2.0–2.5(3.0) 50 5/5
Microbasic p-mastigophores (J) 17.0–22.0(23.0) × 4.0–7.0 50 5/5
Large basitrichs (K) 27.0–34.0(36.0) × (3.0)4.0–6.0(7.0) 50 5/5
Abbreviations: N, the number of specimens for which the type of cnida was present to the total number of specimens examined; n, the number of
cnida capsules measured in this study. Refer to figure 11 for illustrations corresponding to the letters in parentheses.

of red, with both species having adhesive verrucae on 2016); and 3) differing ecology, as M. fautinae sp. nov.
its distal most end, these two species occupy different does not burrow deeply into the substratum with its
habitats. Macrodactyla fautinae sp. nov. tends to occur column completely buried as P. singaporensis does
in silty, sandy areas, among seagrasses meadows of (England 1987; Fautin and Tan 2016); nor, does it occur
the middle intertidal zones, while An. buddemeieri in dense clusters like An. handi, (Dunn 1978; Fautin
occurs underneath rocks in the upper intertidal zone et al. 2009) but rather often occurs singly. For these
(Fautin 2005; Fautin et al. 2009). While the size of a reasons, M. fautinae sp. nov. cannot be confused for the
sea anemone may not be a good character to distinguish other two species.
species (Stephenson 1922; Fautin et al. 2008), we have
consistently found that adult individuals of M. fautinae
sp. nov. are much larger than An. buddemereri (e.g., oral DISCUSSION
disc diameter: 80 mm versus 7 mm, respectively; see
Fautin (2005) for size of An. buddemeieri). Phylogeny of Macrodactyla and D. armata
In occupying the same habitat types (i.e., silty and
sandy substratum), and having translucent, patterned In this study, we tested the relationship between M.
tentacles, M. fautinae sp. nov. may be mistaken for aspera and D. armata, due to their strong resemblance
either An. handi, or P. singaporensis in the field. to each other in having tentacles densely covered
However, M. fautinae sp. nov. can be immediately with visible nematocyst batteries. Our phylogeny
distinguished from these other two species by using the reconstruction did not recover M. aspera and D. armata
following features: 1) a pair of conspicuous pink dots, as a clade. Instead, both formed two distinct groups:
each marking a siphonoglyph (Fig. 8A, B) – a feature M. aspera and the new species, M. fautinae sp. nov.,
that is absent in both An. handi and P. singaporensis; were recovered as a clade, while D. armata was closely
2) a light coloured column with bright pink splotches, related to the actinodendrids (i.e., A. arboreum and Ac.
which differs from the consistently dull and darkly- haeckeli; Fig. 1).
coloured columns of An. handi and P. singaporensis Morphological and ecological evidence supported
(England 1987; Fautin et al. 2009; Fautin and Tan our findings that Macrodactyla species and D. armata
Fig. 11. Cnidae of Macrodactyla fautinae sp. nov. A, spirocyst. B, large spirocyst. C, basitrich. D, large basitrich. E, basitrich. F, basitrich. G,
basitrich. H, microbasic p-mastigophore. I, small basitrich. J, microbasic p-mastigophore. K, large basitrich. Scale bar = 10 µm. Refer to table 3 for
key to letters.

belong to two distinct groups. Anatomically, D. armata (Daly et al. 2017). To determine if D. armata is truly
lacks adhesive verrucae on its column, while these closely related or belongs to the family Actinodendridae
are present on M. aspera and M. fautinae sp. nov. will require further thorough taxonomic and genetic
In both Macrodactyla species, the sphincter muscle work that is beyond the scope of this present study.
is circumscribed in form, while this musculature is
diffused in D. armata (Carlgren 1949). Evidence Macrodactyla aspera as a potential threat to
supporting the close phylogenetic relationship of both public safety
macrodactylid species (i.e., M. aspera and M. fautinae
sp. nov.) have been discussed earlier in this study (see In bearing visible nematocyst batteries, and having
genus entry of Macrodactyla, above). large nematocysts in its tentacles (i.e., undischarged
Together with our molecular findings and some capsules of length > 50 µm), M. aspera may be capable
evidence present in published taxonomic accounts of delivering a painful sting on contact. Dolfeinia
of the Actinodendridae species targeted here, these armata is another, similar example: it bears visible
strongly suggest that D. armata may be actually more nematocyst batteries on its tentacles, and have basitrichs
closely related to actinodendrids than to Actiniidae, that are > 50 µm (e.g., up to 76 µm; see Carlgren 1945).
the family in which Dofleinia is currently placed. Our Other sea anemones that have been reported to be a
proposition here agrees with Rowlett (2020: 292), who threat to public health, such as Phyllodiscus semoni
also suggested the close relatedness of Dofleinia to and members of Actinodendridae also bear specialised
actinodendrids, based on similarities in the ecology and structures along their tentacles that harbour a dense
morphology of these animals. reservoir of nematocysts (e.g., nematocyst batteries)
Ecologically, like D. armata, actinodendrids are and cnidae larger in size (for sizes, see Carlgren
also well-known to be able to inflict painful stings upon 1945; Fautin et al. 2009; Hoeksema and Crowther
contact; they are popularly referred to as ‘Hell’s Fire 2011). Following this line of logic, we suggest that
Anemones’ (Ardelean and Fautin 2004; Erhardt and sea anemone species that may inflict painful stings are
Knop 2005; Fautin et al. 2009). Both D. armata and ones that usually bear specialised structures on their
actinodendrids are hosts to anemone shrimps (Bruce tentacles, and/or have large nematocysts; M. aspera
1969 2005; Fransen 1989). Additionally, Rowlett (2020: may also be a potential threat to humans. This requires
292) notes that these anemones exhibit, “burrowing further investigation and comparisons with other species
behaviour.” that are not deemed to be a threat but also bear large
Broad morphological similarities are shared nematocysts and specialised structures on their tentacles
between D. armata and these actinodendrids, (e.g., Haloclava hercules Izumi, 2021).
particularly: i) in having a smooth column without
any visible structures (e.g., adhesive verrucae, etc.) Importance of re-examining name-bearing type
(Carlgren 1949), and ii) in bearing regions of dense specimens
nematocyst clusters along their tentacles (for a brief
list of other morphological similarities, see Rowlett In this study, we re-described M. aspera using new
2020: 292). Pertaining to i), while assuming M. aspera morphological data (i.e., living appearance, cnidom)
to be a morphological variant of D. armata, Rowlett gathered not only from fresh and voucher specimens, but
(2020: 292) posited that the presence of verrucae is a also from the name-bearing specimen of M. aspera [= C.
diagnostic trait of Dofleinia, despite conceding that this aspera]. This was necessary to ascertain that established
trait was not found on the holotype of its type species diagnostic characters reported for Macrodactyla (e.g.,
(“Dofleinia… having… adhesive verrucae… strangely Stephenson 1921 1922; Carlgren 1949; Dunn 1981)
absent from many specimens (including the type)”). In were truly definitive of the genus. Here, we found that
light of our molecular findings, and the possibility of the complete mesentery numbers and arrangement
Dofleinia being related to actinodendrid anemones, we present in the lectotype differ from previous accounts
propose that the presence of adhesive verrucae is not a that were published. Our findings were not unexpected;
diagnostic trait of Dofleinia. Regarding ii), we note that long-accepted diagnostic characters of a taxon are often
this feature does differ in appearance between D. armata disputed when new observations arise from a careful re-
and the actinodendrids (for an example of acrospheres examination of name-bearing types. Thorough treatment
of an actinodendrid, see Ardelean and Fautin 2004: 192, involving a comprehensive study of name-bearing types
Fig. 3). Rowlett (2020: 292) suggested that this feature will be needed to resolve the taxonomic challenges
may be homologous between these anemones. At for many groups of sea anemones. Members of the
present, the taxonomy of Actiniidae is in chaos and taxa superfamily Actinioidea are taxonomically unresolved
within this family have been found to be polyphyletic (Rodríguez et al. 2014; Titus et al. 2019) and will

benefit from this detailed approach. Recent efforts have Museum of Natural History, Tel Aviv University, Tel
demonstrated such an approach is feasible and fruitful Aviv (Israel); and Western Australian Museum, Perth
(e.g., Fautin and Tan 2016; Yap et al. 2019 2021). (Australia). We are grateful to Mr Lok Kok Sheng for
permission to use his photograph in this publication.
We thank Professor Peter Ng Kee Lin for his advice
CONCLUSIONS on nomenclatural matters on the new species. We also
thank both reviewers of this manuscript for providing
In this study, morphological and molecular constructive feedback and spotting typographical
evidence were integrated to re-diagnose the genus errors. Computational work for this article was partially
Macrodactyla and redescribe M. aspera. Furthermore, performed on resources of the National Supercomputing
we provided historical, nomenclatural evidence to Centre, Singapore (https://www.nscc.sg). The St
support the continued usage of the genus name. Since John’s Island National Marine Laboratory provided
its first description by Haddon and Shackleton (1893), the facilities necessary for conducting this study.
there have been very few published accounts of M. This Laboratory is a National Research Infrastructure
aspera, despite its wide geographical range across the under the National Research Foundation (NRF), Prime
Indo-West Pacific. The reason for this, we suspect, Minister’s Office, Singapore. This research was also
is that this species may have been misidentified as supported by the NRF, under its Marine Science R&D
D. armata, due to its strong superficial resemblance. Programme (MSRDP-P03 and MSRDP-P38).
Museum vouchers we have examined support this
assertion; some were indeed misidentified as D. armata. Authors’ contributions: NWLY collected fresh
Herein, we provided molecular evidence to demonstrate specimens, performed morphological comparisons
that these two species are distantly related. on museum types and vouchers, extracted, sequenced
Findings from this study add to a growing number and analysed genetic data, drafted and formatted
of sea anemone species recorded from Singapore. this manuscript. MLM examined additional WAM
Previous studies have already documented 37 species to specimens, provided 28S sequences for the initial
occur on the shores and islands of this city-state (e.g., phylogeny analyses and drafted the manuscript. ZBRQ
England 1987; Fautin et al. 2009; Yap et al. 2021). sequenced and analysed the genetic data and had
Affirming the identities of these sea anemones primes drafted the manuscript. RT collected fresh specimens,
new avenues where more Indo-West Pacific species may photographed them and provided habitat data on
be included in future research. these animals. KST had guided NWLY on this study,
providing inputs to descriptions and nomenclature
Acknowledgments: This work and the new matters, and had drafted the manuscript. DH had guided
species name were registered with ZooBank under NWLY on this study, analysed the genetic data and
urn:lsid:zoobank.org:pub:560079AC-51E0-448F- drafted the manuscript.
8AAE-449058A9CBF8. We thank the efforts of
Singapore’s ‘Anemone Army’; many of the specimens Competing interests: The authors declare that they
featured in this study were found or collected by these have no conflict of interest.
citizen scientists. Collections of sea anemones were also
made during the Comprehensive Marine Biodiversity Availability of data and materials: Specimens
Survey (CMBS, 2010–2015) and associated workshops documented are kept in natural history museums
led by the National Parks Board Singapore (NParks) and as stated in the paper, genetic sequences and
National University of Singapore (NUS). The CMBS supplementary materials were deposited into GenBank
workshops were supported by generous contributions and Zenodo respectively.
from Asia Pacific Breweries Singapore, Care-for-
Nature Trust Fund, Keppel Care Foundation, Shell Consent for publication: Not applicable.
Companies in Singapore and The Air Liquide Group.
We also thank the following institutions that have Ethics approval consent to participate: Not
granted us permission to examine type material and applicable.
voucher specimens: Lee Kong Chian Natural History
Museum, National University of Singapore, Singapore
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Supplementary materials indicated at the branches as ML/BI. Only bootstrap

values > 50 and posterior probability > 0.8 are shown,
Table S1. Sea anemones species included in our values less than these limits are denoted by a dash (-).
phylogenetic analyses. Information on voucher (download)
specimens and associated museum catalogue numbers
are provided here, if available. Dashes denote that Fig. S4. Maximum likelihood phylogram (RAxML) of
such information is absent. Accession numbers in bold 12S sequences. Bootstrap values (≥ 50) are indicated on
denotes new sequences to this study. Abbreviations: branches. (download)
AMNH, American Museum of Natural History, New
York; CAS, California Academy of Sciences, San Fig. S5. Maximum likelihood phylogram (RAxML) of
Francisco; Cat. nos., Catalogue numbers; KUNHM, 16S sequences. Bootstrap values (≥ 50) are indicated on
University of Kansas Natural History Museum; ZRC, branches. (download)
Zoological Records Collection, Lee Kong Chian Natural
History Museum, Singapore. (download) Fig. S6. Maximum likelihood phylogram (RAxML) of
28S sequences. Bootstrap values (≥ 50) are indicated on
Fig. S1. Map of Singapore where specimens of both branches. (download)
Macrodactyla aspera (Haddon & Shackleton, 1893)
and Macrodactyla fautinae sp. nov. were collected for Fig. S7. Maximum likelihood phylogram (RAxML) of
this study: 1, Pulau Ubin OBS Camp (1°25'07.7"N, cox3 sequences. Bootstrap values (≥ 50) are indicated
103°55'44.3"E); 2, Pasir Ris (1°22'56.45"N, on branches. (download)
103°57'3.88"E); 3, Changi Region beaches [this
area encompass the following localities that are in
very close proximity to each other: Changi beach,
Changi beach near Carpark 4; Changi Point beaches;
Changi Point SAF Chalet beach; Changi Point Ferry
Terminal beach] (1°23'33.34"N, 103°59'20.94"E);
4, Pulau Sekudu (1°24'19"N, 103°59'17"E); 5, Chek
Jawa (1°24'25"N, 103°59'23"E); 6, Beting Bronok
(1°26'13.00"N, 104°02'58.00"E); 7, Changi beach
near Carpark 7 (1°22'26.03"N, 104°0'24.79"E); 8,
Changi East next to Changi water Reclamation Plant
(1°18'45"N, 104°00'31"E); 9, Cyrene Reef (Terumbu
Pandan) (1°15'28"N, 103°45'19"E). White circles
represent M. aspera, while stars (both black and white)
denote the occurrences of M. fautinae sp. nov., with the
single black star indicating the type locality of this new
species. Numbers not accompanied by any symbols
indicate the presence of both species occurring at the
same site. See main-text for full details. (download)
Fig. S2. Distribution of Macrodactyla aspera (Haddon

& Shackleton, 1893) and Macrodactyla fautinae sp.
nov. 1. Murray Islands, Torres Straits. 2. Noumea, New
Caledonia. 3. Perth, Western Australia. 4. Singapore.
5. Eilat, Israel. Symbols beside each number represent
the occurrence of the sea anemones: circles being M.
aspera, stars represents M. fautinae. Symbols that are
coloured black denote the type locality of the species
they represent. See main-text for full details. (download)
Fig. S3. Maximum likelihood phylogram (RAxML)

of concatenated dataset (i.e., 12S, 16S, 28S and cox3).
Bootstrap resampling values under ML, and posterior
probability values of Bayesian inference (BI), are

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Zoological Studies 62:29 (2023)

Taxonomy and Molecular Phylogeny of the

Received 12 August 2022 / Accepted 28 April 2023 / Published 6 July 2023

© 2023 Academia Sinica, Taiwan 1

BACKGROUND greatly impedes further research on these animals.

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

Pairwise genetic distance range

M. aspera M. aspera M. aspera D. armata

Mitochondrial cox3 5.3–6.5% 1.7–2.0% 3.0–3.6% 1.0–1.5%

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

are absent in D. armata; the absence of verrucae is

© 2023 Academia Sinica, Taiwan

Lectotype MZC.I.33665 Vouchers

Tentacles Spirocysts (A) - - (17.0)20.0–30.3 × 1.5–2.5 40 4/4

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

whereas that of M. fautinae sp. nov. is entirely cream-

Tissue Cnidae Range length × width (µm) n N

Tentacles Spirocysts (A) 15.0–27.5 × 2.0–3.0 53 5/5

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

© 2023 Academia Sinica, Taiwan

Supplementary materials indicated at the branches as ML/BI. Only bootstrap

Fig. S2. Distribution of Macrodactyla aspera (Haddon

Fig. S3. Maximum likelihood phylogram (RAxML)

© 2023 Academia Sinica, Taiwan

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