A lungless frog discovered on Borneo

Abstract

The evolution of lunglessness in tetrapods (amphibians, reptiles, birds, and mammals) is an exceedingly rare event. So far lunglessness is known to occur only in amphibians, in particular two families of salamanders 1 and 2 and a single species of caecilian [3]. Here, we report the first case of complete lunglessness in a frog, Barbourula kalimantanensis, from the Indonesian portion of Borneo (Figure 1A). Previously only known from two specimens 4 and 5, a recent expedition to central Kalimantan on Borneo rediscovered two new populations of this enigmatic aquatic frog (Figure 1B,C). This allowed for a more comprehensive assessment of the species' ecology and anatomy that led to the discovery of its lack of lungs. Loss of lungs in Amphibia is most likely due to their evolutionary history at the interface between aquatic and terrestrial habitats and their ancient ability to respire through the skin [5].

Full text

Current Biology Vol 18 No 9
R374
x
= 38.3 mm and in mass from 2.2 to
13.5 g,
x
= 6.5 g). However, we did
locate a glottis and lungs in a specimen
of the only other species in the genus,
Barbourula busuangensis, and another
frog species, Rana catesbeiana
(Figure 2). With no evidence of any lung
tissue and no glottis, B. kalimantanensis
is thus the first species of frog reported
to be lungless.
Among tetrapod vertebrates,
lunglessness has only evolved in
the amphibians: many salamander
species (two species in the family
Hynobiidae, genus Onychodactylus
[2], and more than 350 species in
the family Plethodontidae [1]) as well
as a single species of caecilian (the
other order of amphibians) [3] are
lungless. Thus, the complete loss of
lungs in tetrapods is a particularly
rare evolutionary event. The loss of
lungs is a reversal of one of the most
important physiological adaptations
for terrestrial life and has probably only
evolved independently three times. The
discovery of lunglessness in a secretive
Bornean frog species, supports the
idea that lungs are a malleable trait in
the Amphibia, the sister group to the
rest of the living tetrapods. Amphibians
may be more prone to lunglessness
since they are known to be able to
A lungless frog
discovered on
Borneo
David Bickford1,*, Djoko Iskandar2,
and Anggraini Barlian2
The evolution of lunglessness in
tetrapods (amphibians, reptiles, birds,
and mammals) is an exceedingly rare
event. So far lunglessness is known to
occur only in amphibians, in particular
two families of salamanders [1,2] and
a single species of caecilian [3]. Here,
we report the first case of complete
lunglessness in a frog, Barbourula
kalimantanensis, from the Indonesian
portion of Borneo (Figure 1A). Previously
only known from two specimens [4,5], a
recent expedition to central Kalimantan
on Borneo rediscovered two new
populations of this enigmatic aquatic
frog (Figure 1B,C). This allowed for a
more comprehensive assessment of
the species’ ecology and anatomy that
led to the discovery of its lack of lungs.
Loss of lungs in Amphibia is most likely
due to their evolutionary history at the
interface between aquatic and terrestrial
habitats and their ancient ability to
respire through the skin [5].
Despite multiple attempts to locate
more individuals of B. kalimantanensis,
prior to 2007, only two specimens of
this frog species were known to science
[4,5]. In August 2007, we visited the
type locality near Nanga Pinoh, Western
Kalimantan (0° 44’ S; 111° 40’ E) but
found that illegal gold mining had
destroyed all suitable habitats in the
vicinity. The originally cool, clear, fast-
flowing rivers are now warm and turbid.
Water quality around the type locality
is no longer suitable for the species,
but we were able to discover two new
populations of B. kalimantanensis
upstream of the type locality.
We established the lunglessness of
B. kalimantanensis specimens through
dissections and histological sections
of the anterior portion of the coelom
(around the heart) that revealed a
membrane lining the thoracic cavity,
but no evidence of lungs. In all other
frogs, there is a protected opening
to the airway (the glottis) as the oral
cavity narrows to form the esophagus.
We found no such opening during
dissections of eight specimens of
B. kalimantanesis (ranging in snout-vent
length from 26.9 to 50.5 mm,
readily utilize other methods for gas
exchange, namely cutaneous, gills,
buccopharyngeal and perhaps cloacal
(all thin-membrane) gas exchange
outside of the lungs [6,7].
Respiration determines much of
an organism’s inherent biological
limits and life history. Hence, the
evolution and ecology of lunglessness
is a complex physiological
development entailing many
different mechanisms, possible
explanations, and evolutionary and
developmental pathways. Trade-
offs among kinematic and muscular
performance, buoyancy, and
metabolic rate somehow reach an
evolutionary and ecological balance.
In B. kalimantanensis, this balance
leads to loss of lungs as the main
respiratory surface for gas exchange.
B. kalimantanensis is presumably an
ectotherm and lives in cold (14–17°C)
fast-flowing (2–5 m/s) water, so loss
of lungs may be an adaptation to
the combination of higher oxygen
content in fast-flowing cold water,
the species’ presumed low metabolic
rate, severe flattening to increase the
surface area of the skin (Figure 1B,C),
and selection for negative buoyancy.
B. kalimantanensis, the only lungless
tetrapod in Southeast Asia, is currently
Figure 1. Habitat and appearance of the lungless frog Barbourula kalimantanensis.
(A) Map of Borneo, showing the Indonesian portion, Kalimantan, in the South-Central part of
the island, and (B) B. kalimantanensis in anterior view, and (C) lateral view showing extreme
flattening of the body.

Magazine
R375
listed as endangered [8] and illegal
gold mining resulting in increased
turbidity and mercury contamination
has severely degraded the type locality
and much of its presumed former range.
Compounding the problem, much of
the surrounding terrestrial habitat is
also under increasing threat from both
legal and illegal logging. Conservation
of this evolutionary enigma needs to be
prioritized and the remaining habitat in
which it can survive needs to be urgently
protected. The evolution, development,
and maintenance of lunglessness in this
frog will become important research foci.
How complete loss of lungs evolves and
under what kind of selective pressures
and genetic mechanisms has been
well debated in salamanders [9,10].
However, these are still open and more
manageable questions for an aquatic
primitive frog. To better understand the
extinction risk and endangered status
of this species, a much more complete
assessment of potential habitats needs
to be surveyed and the exact geographic
range for the species should be mapped.
In addition, virtually nothing is known
about how these frogs reproduce, eat
and escape predation. Further studies,
however, may be hampered by the
species’ rarity and endangerment. We
strongly encourage conservation of the
remaining habitats of this species.
Supplemental data
Supplemental data including experimental pro-
cedures are available at http://www.current-
biology.com/cgi/content/full/18/9/R374/DC1
Acknowledgments
We thank Rafe Brown, Rudolf Meier, and two
anonymous reviewers for helpful comments.
The project would not have been successful
without support from Darmawan Liswanto. In
the field, we were assisted immeasurably by
Mistar Kamsi, Umilaela, Angga Rachmansah,
Biofagri A.R., Medi Yansyah, Budi Susilo, Herry
Helmi, Doddy Aryadi, and the staff of the Taman
Nasional Bukit Baka – Bukit Raya. We thank the
Forestry Department, Sintang, Kalimantan Barat
for permission to conduct research in the area
and the Ministry of Education of the Republic
of Singapore for providing funding under Grant
#R-154-000-270-112.
References
1. Min, M.S., Yang, S.Y., Bonett, R.M., Vieites, D.R.,
Brandon, R.A., and Wake, D.B. (2005). Discovery
of the first Asian plethodontid salamander.
Nature 435, 87–90.
2. Dunn, E.R. (1923). The salamanders of the family
Hynobiidae. Proc. Amer. Acad. Arts Sci. 58,
445–523.
3. Nussbaum, R.A., and Wilkinson, M. (1995). A new
genus of lungless tetrapod: A radically divergent
caecilian (Amphibia: Gymnophiona). Proc. R.
Soc. Lond. B 261, 331–335.
4. Iskandar, D.T. (1978). A new species of
Barbourula: First record of a discoglossid from
Borneo. Copeia 1978, 564–566.
5. Iskandar, D.T. (1995). Note on the second
specimen of Barbourula kalimantanensis
(Amphibia: Anura: Discoglossidae). Raffles Bull.
Zool. 43, 309–311.
6. Cox, C.B. (1967). Cutaneous respiration and the
origin of the modern Amphibia. Proc. R. Soc.
Lond. B. 178, 37–47.
7. Duellman, W., and Trueb, L. (1986). Biology of the
Amphibia. (Baltimore: Johns Hopkins University
Press).
8. IUCN, Conservation International, and
NatureServe. (2006). Global Amphibian
Assessment. <www.globalamphibians.org>.
9. Wilder, I.W., and Dunn, E.R. (1920). The
correlation of lunglessness in salamanders with
a mountain brook habitat. Copeia 84, 63–68.
10. Ruben, J.A., and Boucot, A.J. (1989). The
origin of the lungless salamanders (Amphibia:
Plethodontidae). Am. Nat. 134, 161–169.
1National University of Singapore, 14
Science Drive 4, 117543 Singapore. 2Institut
Teknologi Bandung, 10 Jalan Ganesa,
Bandung, 40132 Java, Indonesia.
*E-mail: rokrok@nus.edu.sg
Figure 2. Anatomy of lunglessness.
Comparison of (A) typical frog mouth and pharynx (Rana catesbeiana), showing glottis
(circled), tongue, and esophageal opening, and (B) B. kalimantanensis showing tongue, no
glottis (circled), and an enlarged esophageal opening leading directly to the stomach.
Role of fungi in the
biogeochemical fate
of depleted uranium
Marina Fomina1, John M. Charnock2,
Stephen Hillier3, Rebeca Alvarez4,
Francis Livens4
and Geoffrey M. Gadd1,*
The testing of depleted uranium (DU; a
97.25% U:0.75% Ti alloy) ammunition
and its use in recent war campaigns in
Iraq (1991 and 2003) and the Balkans
(1995 and 1999) has led to dispersion
of thermodynamically unstable DU
metal into the environment [1–3].
Although less radioactive, DU has
the same chemotoxicity as natural
uranium and poses a threat to human
populations [1]. Uranium tends to
form stable aqueous complexes and
precipitates with organic ligands [4],
suggesting that living organisms could
play an important role in geochemical
transformations and cycling. Fungi
are one of the most biogeochemically
active components of the soil
microbiota [5], particularly in the
aerobic plant-root zone. Although the
mutualistic symbiotic associations
(mycorrhizas) of fungi with plants
are particularly important in mineral
transformations [5], fungal effects on
metallic DU have not been studied.
Here, we report that free-living and
plant symbiotic (mycorrhizal) fungi can
colonize DU surfaces and transform
metallic DU into uranyl phosphate
minerals.
Fungal interactions with DU were
studied in microcosms simulating
a heterogeneous environment
(Figure S1A in Supplemental Data,
published with this article online).
All tested fungi exhibited high DU
tolerance and were able to colonize
DU surfaces, forming moisture-
retaining mycelial biofilms (Figure
S1A–D). The fungi also often formed
cord-like mycelial structures through
aggregation of longitudinally aligned
hyphae (Figures 1A,B and S1F,G),
commonly interpreted as a survival
response to metal stress [6].
DU coupons (triangular sectors of
DU alloy of approximate dimensions
15 mm x 15 mm x 11 mm, and 5 mm
height, and approximately 6.5–8.5 g in
weight) in the microcosms underwent
aerobic corrosion forming black
and yellow decomposition products