On the distribution and conservation of two “Lost World” tepui summit endemic frogs, Stefania ginesi Rivero, 1968 and S. satelles Señaris, Ayarzagüena, and Gorzula, 1997

Abstract

It has been suggested that the inability to migrate in response to climate change is a key threat to tepui summit biota. Tepui summit organisms might thus seriously be threatened by global warming, and there is an urgent need to accurately evaluate their taxonomic status and distributions. We investigated phylogenetic relationships among several populations of Stefania ginesi and S. satelles, two endemic species reported from some isolated tepui summits, and we examined their IUCN conservation status. Molecular phylogenetic analysis and preliminary morphological assessment indicate that both species are actually restricted to single tepui summits and that five candidate species are involved under these names. We advocate upgrading the conservation status of S. ginesi from Least Concern to Endangered, and that of S. satelles from Near Threatened to Endangered.

Full text

5
Amphib. Reptile Conserv. April 2016 | Volume 10 | Number 1 | e115
Amphibian & Reptile Conservation
10(1) [Special Section]: 5–12 (e115).
On the distribution and conservation of two “Lost World”
tepui summit endemic frogs, Stefania ginesi Rivero, 1968 and
S. satelles Señaris, Ayarzagüena, and Gorzula, 1997
1,3Philippe J. R. Kok, 1,4Valerio G. Russo, 1,5Sebastian Ratz, and 2,6Fabien Aubret
1Amphibian Evolution Lab, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, BELGIUM 2Station d’Ecologie Expérimentale du CNRS à
Moulis, USR 2936, 09200 Moulis, FRANCE
Abstract.—It has been suggested that the inability to migrate in response to climate change is a key
threat to tepui summit biota. Tepui summit organisms might thus seriously be threatened by global
warming, and there is an urgent need to accurately evaluate their taxonomic status and distributions.
We investigated phylogenetic relationships among several populations of Stefania ginesi and
S. satelles, two endemic species reported from some isolated tepui summits, and we examined
their IUCN conservation status. Molecular phylogenetic analysis and preliminary morphological
assessment indicate that both species are actually restricted to single tepui summits and that ve
candidate species are involved under these names. We advocate upgrading the conservation status
of S. ginesi from Least Concern to Endangered, and that of S. satelles from Near Threatened to
Endangered.
Key words. Endangered species, Hemiphractidae, IUCN, molecular phylogenetics, molecular taxonomy, Venezuela
Citation: Kok PJR, Russo VG, Ratz S, Aubret F. 2016. On the distribution and conservation of two “Lost World” tepui summit endemic frogs, Stefania
ginesi Rivero, 1968 and S. satelles Señaris, Ayarzagüena, and Gorzula, 1997. Amphibian & Reptile Conservation 10(1): 5–12 (e115).
Copyright: © 2016 Kok et al. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-
NoDerivatives 4.0 International License, which permits unrestricted use for non-commercial and education purposes only, in any medium, provided
the original author and the o󰀩cial and authorized publication sources are recognized and properly credited. The o󰀩cial and authorized publication
credit sources, which will be duly enforced, are as follows: o󰀩cial journal title Amphibian & Reptile Conservation; o󰀩cial journal website <amphibian-
reptile-conservation.org>.
Received: 08 March 2016; Accepted: 29 March 2016; Published: 12 April 2016
Correspondence. Email: 3Philippe.Kok@vub.ac.be (Corresponding author); 4valerio.giovanni.russo@gmail.com;
5Sebastian.Ratz@vub.ac.be; 6faubret@gmail.com
O󰀩cial journal website:
amphibian-reptile-conservation.org
Introduction
The frog genus Stefania (Hemiphractidae) is endemic
to an iconic South American biogeographical region
named “Pantepui” (Mayr and Phelps 1967; McDiarmid
and Donnelly 2005) (Fig. 1). Pantepui, often referred to
as the “Lost World” because of Arthur Conan Doyle’s
famous novel (1912), lies in the western Guiana Shield.
The region harbors numerous isolated Precambrian
sandstone tabletop mountains more formally known as
“tepuis” (Fig. 2). Although Pantepui was initially re-
stricted to tepui slopes and summits above 1,500 m el-
evation (Mayr and Phelps 1967; Rull and Nogué 2007),
Steyermark (1982), followed by Kok et al. (2012) and
Kok (2013a), expanded the original denition of Pan-
tepui to include the intervening Pantepui lowlands (200-
400 m asl) and uplands (400-ca. 1,200 m asl) in order
to better reect the biogeography and biotic interactions
in the area (Kok 2013a). The genus Stefania currently
includes 19 species, 15 of which are restricted to tepui
slopes or summits (Duellman 2015; Frost 2015). Stefa-
nia species are direct-developers (eggs and juveniles car-
ried on the back of the mother) and occupy various types
of habitats from lowland rainforest to tepui bogs (Kok
2013a; Schmid et al. 2013; Duellman 2015). The genus
Stefania was erected by Rivero (1968) to accommodate
Cryptobatrachus evansi and a few related new species all
morphologically divergent from other Cryptobatrachus.
Shortly later, Rivero (1970) recognized two species-
groups within Stefania: the evansi group including spe-
cies having the head longer than broad and found in the
lowlands and uplands of Pantepui, and the goini group
including species having the head broader than long and
found in the highlands of Pantepui. Kok et al. (2012),
followed by Castroviejo et al. (2015), showed that, based
on molecular data, these groups are actually not recip-
rocally monophyletic. A complete molecular phyloge-
netic analysis of the genus Stefania is still lacking, and

6Amphib. Reptile Conserv. April 2016 | Volume 10 | Number 1 | e115
Kok et al.
relationships between many species or populations are
unknown. Likewise, the exact distribution of some tepui
summit species is uncertain (e.g., Gorzula and Señaris
1999). Among these, two tepui summit endemic Stefania
species are known from several isolated tepui summits:
Stefania ginesi Rivero, 1968, which is reported from six
tepuis in the Chimantá massif (Chimantá-tepui, Amurí-
tepui, Abakapá-tepui, Churí-tepui, Akopán-tepui, and
Murei-tepui; Señaris et al. 1997; Gorzula and Señaris
1999; Barrio-Amorós and Fuentes 2012; Fig. 1), and Ste-
fania satelles Señaris, Ayarzagüena, and Gorzula, 1997,
which has a highly disjunct distribution, being reported
from Aprada-tepui (in the Aprada Massif), Angasima-
tepui, and Upuigma-tepui (two southern outliers of the
Chimantá massif), and from Murisipán-tepui and Ka-
markawarai-tepui (in the Los Testigos Massif, north of
the Chimantá massif) (Señaris et al. 1997; Gorzula and
Señaris 1999; Fig. 1). Stefania ginesi is listed as Least
Concern (LC) by the International Union for Conserva-
tion of Nature (IUCN) (Stuart et al. 2008) and S. satelles
is listed as Near Threatened (NT) (Stuart et al. 2008).
However, preliminary data suggest that their respec-
tive distributions could be more restricted than initially
thought because more than two species could be involved
under these names (the authors, unpublished; see also Se-
ñaris et al. 2014 regarding the distribution of S. ginesi).
Herein we used molecular phylogenetics to investigate
the relationships among three populations of S. ginesi and
four populations of S. satelles. We also aim at providing
a more precise distribution of these two taxa in order to
rene their conservation status. Indeed, tepui ecosystems
are reported as particularly sensitive to global warming
(Nogué et al. 2009), and tepui summit organisms might
be seriously threatened by habitat loss due to upward
displacement (Rull and Vegas-Vilarrúbia 2006; see also
below). Likewise, climate envelope distribution models
of tepui ecosystems based on future scenarios show that
potential distributions become drastically smaller under
global warming (Rödder et al. 2010). Species restricted
to tepui summits are thus clearly at risk of extinction, and
there is an urgent need to evaluate their exact taxonomic
status and precise distribution.
Materials and Methods
Tissue sampling and molecular data
We combined available GenBank sequences of Stefania
ginesi and S. satelles for fragments of the mitochondrial
16SrRNA gene (16S) and the protein-coding mitochon-
drial gene NADH hydrogenase subunit 1 (ND1) with 40
novel DNA sequences of Stefania ginesi and S. satelles:
nine of fragments of 16S, ve of ND1, 13 of the nuclear
recombination activating gene 1 (RAG1), and 13 of the
nuclear CXC chemokine receptor type 4 gene (CXCR4).
We combined this dataset with DNA sequences of four
additional members of the genus Stefania from out-
side the studied area (three species from east of the Río
Caroní: S. scalae, an upland species, S. riveroi and S.
schuberti, two highland species; and one highland spe-
Fig. 1. Left: Map of Pantepui and its location within South America (inset); the thick blue line indicates the Río Caroní. Right: Map
of the area under study showing localities mentioned in the text (yellow dots represent known localities of occurrence of Stefania
satelles, white dots represent known localities of occurrence of Stefania ginesi). Numbers indicate sampled localities and Roman
numerals indicate unsampled localities, as follows: (1) Aprada-tepui, Venezuela; (2) Murisipán-tepui, Venezuela; (3) Upuigma-
tepui, Venezuela; (4) Angasima-tepui, Venezuela; (5) Abakapá-tepui, Venezuela; (6) Chimantá-tepui, Venezuela; (7) Amurí-tepui,
Venezuela; (i) Kamarkawarai-tepui, Venezuela; (ii) Murei-tepui, Venezuela; (iii) Churí-tepui, Venezuela; (iv) Akopán-tepui, Ven-
ezuela.

7Amphib. Reptile Conserv. April 2016 | Volume 10 | Number 1 | e115
“Lost World” tepui summit endemic frogs, Stefania ginesi and S. satelles
cies from west of the Río Caroní: S. riae; in total 16 novel
sequences), and with Fritziana ohausi, member of the
clade sister to Stefania (Castroviejo et al. 2015), which
was selected as outgroup (see Table 1). Novel sequences
have been catalogued in GenBank under the accession
numbers KU958582-958637.
Total genomic DNA was extracted and puried using
the Qiagen DNeasy® Tissue Kit following manufactur-
er’s instructions. Fragments of 16S (ca. 550 base pairs
[bp]), of ND1 (ca. 650 bp), and of RAG1 (ca. 550 bp)
and CXCR4 (ca. 625 bp) were amplied and sequenced
using the primers listed in Kok et al. (2012) and Biju and
Bossuyt (2003) under previously described PCR condi-
tions (Biju and Bossuyt 2003; Roelants et al. 2007; Van
Bocxlaer et al. 2010). PCR products were checked on
a 1% agarose gel and were sent to BaseClear (Leiden,
The Netherlands) for purication and sequencing. Chro-
matograms were read using CodonCode Aligner 5.0.2
Fig. 2. Typical Pantepui landscape. Photograph taken on 8th June 2012 from the summit of Upuigma-tepui, showing Angasima-tepui
on the left and Akopán-tepui and Amurí-tepui on the right. Note stretches of savannah mainly caused by anthropogenic res. Photo
PJRK.
Voucher 16S ND1 RAG1 CXCR4 Genus Species Locality Country Coordinates Elevation (m)
IRSNB16724 JQ742191 JQ742362 KU958600 KU958619 Stefania scalae Salto El Danto Venezuela N 5°57’52” W 61°23’31” 1208
Uncatalogued JQ742172 JQ742343 KU958601 KU958620 Stefania riae Sarisariñama-tepui Venezuela N 4°41’ W 64°13’ ca. 1100
IRSNB15703 JQ742177 JQ742348 KU958602 KU958621 Stefania riveroi Yuruaní-tepui Venezuela N 5°18’50” W 60°51’50” 2303
IRSNB15716 JQ742178 JQ742349 KU958603 KU958622 Stefania riveroi Yuruaní-tepui Venezuela N 5°18’50” W 60°51’50” 2303
IRSNB16725 JQ742173 JQ742344 KU958604 KU958623 Stefania “ginesi” Abakapá-tepui Venezuela N 5°11’23” W 62°17’52” 2137
IRSNB16726 JQ742174 JQ742345 KU958605 KU958624 “ginesi” “ginesi” Abakapá-tepui Venezuela N 5°11’07” W 62°17’21” 2209
IRSNB15839 JQ742175 JQ742346 KU958606 KU958625 Stefania “satelles” Angasima-tepui Venezuela N 5°02’36” W 62°04’51” 2122
IRSNB15844 JQ742176 JQ742347 KU958607 KU958626 Stefania “satelles” Angasima-tepui Venezuela N 5°02’36” W 62°04’51” 2122
IRSNB16727 KU958582 KU958593 KU958608 KU958627 Stefania “satelles” Upuigma-tepui Venezuela N 5°05’10” W 61°57’32” 2134
IRSNB16728 KU958583 —KU958609 KU958628 Stefania satelles Aprada-tepui Venezuela N 5°24’39” W 62°27’00” 2551
IRSNB16729 KU958584 —KU958610 KU958629 Stefania satelles Aprada-tepui Venezuela N 5°24’43” W 62°27’03” 2576
IRSNB16730 KU958585 KU958594 KU958611 KU958630 Stefania “ginesi” Amurí-tepui Venezuela N 5°08’34” W 62°07’08” 2215
IRSNB16731 KU958586 KU958595 KU958612 KU958631 Stefania “ginesi” Amurí-tepui Venezuela N 5°08’35” W 62°07’08” 2213
IRSNB16732 KU958587 KU958596 KU958613 KU958632 Stefania schuberti Auyán-tepui Venezuela N 5°45’56” W 62°32’25” 2279
IRSNB16733 KU958588 KU958597 KU958614 KU958633 Stefania schuberti Auyán-tepui Venezuela N 5°45’56” W 62°32’25” 2279
IRSNB16734 KU958589 KU958598 KU958615 KU958634 Stefania “satelles” Murisipán-tepui Venezuela N 5°52’03” W 62°04’30” 2419
IRSNB16735 KU958590 KU958599 KU958616 KU958635 Stefania “satelles” Murisipán-tepui Venezuela N 5°52’03” W 62°04’30” 2419
IRSNB16736 KU958591 —KU958617 KU958636 Stefania ginesi Chimantá-tepui Venezuela N 5°19’12” W 62°12’07” 2180
IRSNB16737 KU958592 —KU958618 KU958637 Stefania ginesi Chimantá-tepui Venezuela N 5°19’12” W 62°12’07” 2180
MZUSP139225 JN157635 KC844945 KC844991 — Fritziana ohausi n/a Brazil n/a n/a
Table 1. List of Stefania taxa and outgroup used in this study, with localities and GenBank accession numbers. Sequences newly
generated are in boldface. IRSNB = Institut Royal des Sciences Naturelles de Belgique, Belgium; MZUSP = Museu de Zoologia,
Universidade de São Paulo, Brazil.

8Amphib. Reptile Conserv. April 2016 | Volume 10 | Number 1 | e115
Kok et al.
(http://www.codoncode.com/index.htm) and a consensus
sequence was assembled from the forward and reverse
primer sequences. MAFFT version 7 (http://mat.cbrc.
jp/alignment/server/) was used to perform preliminary
alignment using G-INS-i and default parameters. Mi-
nor alignment corrections were made using MacClade
4.08 (Maddison and Maddison 2005). Protein-coding
sequences were translated into amino-acid sequences to
check for unexpected stop codons. Alignment-ambiguous
regions of 16S were excluded from subsequent analyses.
Molecular phylogenetic analyses
The combined 16S + ND1 + RAG1 + CXCR4 dataset
(totalling 2,359 bp after exclusion) was subjected to phy-
logenetic inference using Bayesian analyses. Optimal
partitioning schemes were estimated with PartitionFinder
v1.1.1 (Lanfear et al. 2012) using the “greedy” algorithm,
the “mrbayes” set of models, and the Bayesian Informa-
tion Criterion (BIC) to compare the t of dierent mod-
els. Bayesian posterior probabilities (PP) were used to
estimate clade credibility in MrBayes 3.2.2 (Ronquist et
al. 2012) on the CIPRES Science Gateway V 3.3 (https://
www.phylo.org/, Miller et al. 2010). The Bayesian analy-
ses implemented the best substitution models inferred by
PartitionFinder v1.1.1 partitioned over the dierent gene
fragments, at Dirichlet priors for base frequencies and
substitution rate matrices and uniform priors for among-
site rate parameters. Four parallel Markov chain Monte
Carlo (MCMC) runs of four incrementally heated (tem-
perature parameter = 0.2) chains were performed, with a
length of 20,000,000 generations, a sampling frequency
of 1 per 1,000 generations, and a burn-in correspond-
ing to the rst 1,000,000 generations. Convergence of
the parallel runs was conrmed by split frequency SDs
(<0.01) and potential scale reduction factors (~1.0) for
all model parameters, as reported by MrBayes. All analy-
ses were checked for convergence by plotting the log-
likelihood values against generation time for each run,
using Tracer 1.5 (Rambaut and Drummond 2009). Eec-
tive sample sizes (ESS) largely over 200 were obtained
for every parameter. Results were visualized and edited
in FigTree 1.4.1 (Rambaut 2014).
Results
Stefania ginesi and S. satelles as currently recognized
are recovered non-reciprocally monophyletic (Fig. 3).
Our molecular phylogeny also reveals the occurrence of
ve candidate species (sensu Padial et al. 2010) that have
been misidentied for more than a decade as S. ginesi
(two candidate species) or S. satelles (three candidate
species) (e.g., Señaris et al. 1997; Gorzula and Señaris
1999). Preliminary morphological analyses (in progress)
indicate a few, sometimes subtle, morphological charac-
ters allowing discrimination among these candidate spe-
Fig. 3. Phylogenetic relationships as recovered in the MrBayes analysis (concatenated dataset, 2359 bp), outgroup not shown.
Values at each node represent Bayesian posterior probabilities; asterisks indicate values > 95%. Stefania ginesi sensu stricto, and
S. satelles sensu stricto are highlighted in red. Relation between eye color and tepui summit surface is indicated on the right side of
the gure. Photos PJRK.

9Amphib. Reptile Conserv. April 2016 | Volume 10 | Number 1 | e115
“Lost World” tepui summit endemic frogs, Stefania ginesi and S. satelles
cies and S. ginesi and S. satelles. Our combined results
indicate that S. ginesi sensu stricto is likely restricted to
its type locality, Chimantá-tepui, as we suspect that pop-
ulations from other tepuis in the Chimantá Massif that
were not sampled in this study will prove to be distinct as
well. As for Stefania satelles, the species is restricted to
its type locality, Aprada-tepui.
Discussion and conservation recommendations
We assumed that misidentications were likely due
to a rather conserved external morphology (e.g., head
broader than long, skin strongly granular, absence of
prominent cranial crests) of all tepui summit species pre-
viously identied as Stefania ginesi or S. satelles. This
conserved morphology appears to be symplesiomorphic,
and probably the result of an allopatric non-adaptive ra-
diation (lineage diversication with minimal ecological
diversication, see Rundell and Price 2009). It is, how-
ever, intriguing that two slightly divergent phenotypes (a
“satelles phenotype” with brown eyes and a “ginesi phe-
notype” with blue eyes) evolved independently in each
subclade (see Fig. 3). Interestingly, selection towards one
of these two phenotypes seems closely associated with
the size of the summit surface on which the species occur
(see Fig. 3). The “ginesi phenotype” is found on large
tepui summits (surface > 25 km2) in the central Chimantá
Massif, whereas the “satelles phenotype” is found on
much smaller tepui summits (surface < 5 km2) in the pe-
riphery of the core Chimantá Massif. Disentangling this
phenomenon and the nature of the ecological constraints
possibly involved and their inuence on phenotypic tra-
jectories is beyond the scope of this paper and will be
treated in a separate study.
Most importantly, our results have direct implica-
tions on the conservation status of the populations un-
der study. A complete taxonomic revision of the genus
is in progress, but meanwhile we wish to emphasize the
restricted distributions of all the populations previously
known as Stefania ginesi or S. satelles. Our results argue
for the upgrading of the conservation status of S. gine-
si from LC to Endangered (EN), and that of S. satelles
from NT to EN, based on the same argument recently
developed for other species restricted to the summit of
one or two tepuis, e.g., Pristimantis imthurni and P. jam-
escameroni (Kok 2013b), or P. aureoventris (IUCN SSC
Amphibian Specialist Group 2014), thus in accordance
with criteria B1 a-b (iii) and B2 a-b (iii) of the IUCN
Red List of Threatened Species (IUCN 2014). We indeed
argue that (1) extents of occurrence of S. ginesi and S.
satelles are much less than 5,000 km2 (less than 100 km2
and ve km2, respectively); (2) areas of occupancy of S.
ginesi and S. satelles are much less than 500 km2 (less
than 100 km2 and ve km2, respectively); (3) there is an
inferred and projected decline in the quality of habitat
due to the eects of global warming upon tepui ecosys-
tems, with an expected 2–4 °C increase in temperature
in the region through the next century (IPCC 2007). As
stressed by Nogué et al. (2009) and Rödder et al. (2010),
this rise in temperature will likely cause a decrease in
habitat suitability for tepui biota. In addition, numerous
anthropogenic res in the region (Means 1995; Rull et al.
2013, 2016), coupled with a global rise of temperature,
may cause an up to 10% decrease in precipitation (IPCC
2007) instigating an increase in re range and intensity
(Rull et al. 2013, 2016); and (4) the altitudinal range of
Stefania ginesi and S. satelles allows no vertical migra-
tion in order to avoid these threats. As mentioned by Rull
and Vegas-Vilarrúbia (2006), the inability to migrate to
compensate for the climate change is a key threat to tepui
summit biota.
There is an urgent need to gain a greater understand-
ing of species boundaries and distributions in Pantepui,
especially in Venezuela where the threats are the highest
due to ongoing uncontrolled anthropogenic res (Rull
et al. 2013, 2016). However, it is assumed that an even
greater threat to Pantepui biota is global climate change.
Local actions (such as stopping res), even if necessary,
might only have a limited impact on the long-term sur-
vival of Pantepui organisms. Conservation awareness is
critically important in the area, notably due to the inac-
cessibility of tepui ecosystems where an out of sight, out
of mind eect may have taken place.
This study adds to the many studies now available
demonstrating that estimates of amphibian diversity
based on morphology alone are often misleading. Molec-
ular data have indeed been shown to be of great help in
detecting cryptic species (e.g., Hebert et al. 2004; Vences
et al. 2005; Fouquet et al. 2007; Burns et al. 2008; Fou-
quet et al. 2016). Unfortunately, while everyone seems to
agree that gaining a greater understanding of the world
biodiversity is needed in order to prioritize biodiversity
conservation (e.g., Wilson 2016), the task turns more and
more often into a bureaucratic obstacle course, if not an
impossible mission for scientists working with molecular
data.
Acknowledgments.—PJRK’s work is supported by
a postdoctoral fellowship from the Fonds voor Weten-
schappelijk Onderzoek Vlaanderen (FWO12A7614N).
Many thanks are due to C.L. Barrio-Amorós (Doc Frog
Expeditions, Costa Rica) and C. Brewer-Carías (Caracas,
Venezuela) for the loan of tissue samples. C. Brewer-
Carías also provided invaluable advice and help with lo-
gistics in Venezuela.
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Sebastian Ratz has a Bachelor’s degree in biology from the University of Tübingen, Germany. He currently
works on his Master thesis (phylogeography of the genus Oreophrynella) at the Vrije Universiteit Brussel, Bel-
gium. His main interests focus on the diversity and evolution of Neotropical amphibians.
Philippe J.R. Kok is a Belgian evolutionary biologist and herpetologist. He obtained his Ph.D. in biology at the
Leiden University (The Netherlands) in 2013. He is currently postdoctoral researcher at the Vrije Universiteit
Brussel, Belgium, where he teaches Field Herpetology to the second year Master students. His interests are
eclectic, the main ones being the evolution, systematics, taxonomy, biogeography, and conservation of amphib-
ians and reptiles in the Neotropics, more specically from the Guiana Shield. His work now primarily focuses
on vertebrate evolution in the Pantepui region.
Valerio G. Russo is an Italian herpetologist and naturalist mainly interested in Neotropical and Mediterranean
biodiversity. He obtained his Master’s degree in biology in 2015 at the Vrije Universiteit Brussel (VUB), Bel-
gium, with a thesis on the systematics of the frog genus Stefania. He is now collaborating as an independent
researcher with the Biology Department of the VUB.
Fabien Aubret is a French evolutionary biologist and herpetologist. He completed his Doctoral and Post-
doctoral studies between 2001 and 2008 in Australia (University of Western Australia and University of Syd-
ney). Since 2009, he has been working as a full time researcher for the CNRS (National Centre for Scientic
Research) at the Station of Theoretical and Experimental Ecology (SETE, Moulis, France). Fabien’s research
is mostly empirical, with an experimental backbone, and involves a variety of snake and lizard models. His
research is pluri-disciplinary and involves eco-physiology, phenotypic plasticity, climate change, thermoregula-
tion, and reproductive biology.
Kok et al.