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Rediscovery of the nearly extinct longnose
harlequin frog Atelopus longirostris (Bufonidae) in
Junín, Imbabura, Ecuador
Elicio Eladio Tapia, Luis Aurelio Coloma , Gustavo Pazmiño-Otamendi &
To cite this article: Elicio Eladio Tapia, Luis Aurelio Coloma , Gustavo Pazmiño-Otamendi
& Nicolás Peñafiel (2017) Rediscovery of the nearly extinct longnose harlequin frog Atelopus
longirostris (Bufonidae) in Junín, Imbabura, Ecuador, Neotropical Biodiversity, 3:1, 157-167, DOI:
To link to this article: http://dx.doi.org/10.1080/23766808.2017.1327000
© 2017 The Author(s). Published by Informa
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Published online: 25 May 2017.
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Rediscovery of the nearly extinct longnose harlequin frog Atelopus longirostris (Bufonidae) in
Junín, Imbabura, Ecuador
Elicio Eladio Tapia
, Luis Aurelio Coloma
*, Gustavo Pazmiño-Otamendi
and Nicolás Peñaﬁel
Centro Jambatu de Investigación y Conservación de Anﬁbios, Fundación Otonga, Quito, Ecuador;
Amazónica IKIAM, Tena, Ecuador;
Laboratorio de Biología Molecular, Centro de Investigación de la Biodiversidad y Cambio
Climático, Universidad Tecnológica Indoamérica, Machala y Sabanilla, Quito, Ecuador
(Received 22 November 2016; accepted 27 April 2017)
We report the recent ﬁnding of four adults of Atelopus longirostris, a Critically Endangered species that was last seen in
1989, when catastrophic Atelopus declines occurred. The rediscovery of A. longirostris took place in a new locality,
Junín, 1250–1480 m asl, Provincia Imbabura, Ecuador, on 28–31 March 2016. The four frogs were found in two isolated
small patches of native forest in a fragmented area heavily modiﬁed for agriculture and livestock; one patch protected by
the Junín Community Reserve, and another non-protected private patch near the reserve. We found high prevalence of
Batrachochytrium dendrobatidis (Bd) in the amphibian community of Junín, but A. longirostris tested negative. The ﬁnd-
ing of A. longirostris after 27 years is surprising and ﬁts an apparent pattern of mild conditions that might be promoting
either the recovery or persistence in low numbers of some relict amphibian populations. The frogs are the ﬁrst founders
of an ex situ assurance colony in Jambatu Research and Conservation Center. Expansion of the Junín Community
Reserve is urgently needed to add the currently non-protected patch of forest, where A. longirostris also occurs. The
restoration of the forest in degraded areas between both forest patches and in the related river margins is also necessary.
This restoration will grant the connectivity between both isolated metapopulations and the normal movement of individu-
als to the breeding sites in the Chalguayacu and Junín River basins. The latter should be protected to prevent any kind
of water pollution by the opencast copper exploitation of the mining concession Llurimagua, which is underway.
Atelopus longirostris belongs to a group of at least 29 species of Ecuadorian Atelopus that are critically endangered, 15
of which remain unsighted for at least one decade, and most of them might be extinct. Further synchronous, multidisci-
plinary and integrative research is needed, aiming to understand the most aspects of the biology of species of Atelopus
to support in situ and ex situ conservation actions.
Keywords: Atelopus longirostris; Bufonidae; Ecuador; extinction; rediscovery
Reportamos el reciente hallazgo de cuatro adultos de Atelopus longirostris, una especie en Peligro Crítico, la misma que
fue vista por última vez en 1989, cuando se produjeron declives catastróﬁcos de Atelopus. El redescubrimiento de A.
longisrostris tuvo lugar en una nueva localidad, Junín, 1250–1480 msnm, Provincia de Imbabura, Ecuador, entre el
28–31 de marzo de 2016. Las cuatro ranas se encontraron en dos pequeñas parcelas de bosque natural en un área frag-
mentada y densamente modiﬁcada para agricultura y ganadería, la una parcela forma parte de la Reserva de la Comu-
nidad Junín y la otra está en un área privada no protegida cercana a la reserva. Encontramos alta prevalencia de
Batrachochytrium dendrobatidis (Bd) en la comunidad de anﬁbios de Junín, aunque no se encontró en A. longirostris.
Su hallazgo después de 27 años es sorprendente y se ajusta a un patrón aparente de condiciones benignas que estarían
promoviendo sea la recuperación o persistencia de poblaciones relictas de algunas especies de anﬁbios. Estas ranas son
los primeros fundadores de una colonia de manejo ex situ en el Centro Jambatu de Investigación y Conservación de
Anﬁbios. Se necesita con urgencia la expansión de la Reserva de la Comunidad de Junín para incluir todos los bosques
en donde A. longisrostris habita. Es también necesaria la restauración de los bosques en las áreas destruidas que quedan
entre los parches de bosque y en la rivera de los ríos. Esta restauración garantizará la conectividad entre metapoblaciones
aisladas y también el desplazamiento normal de individuos a los sitios de reproducción en las cuencas de los ríos
Chalguayacu y Junín. Estas cuencas deben ser protegidas para evitar cualquier tipo de contaminación en el agua pro-
ducida por la explotación de cobre a cielo abierto de la concesión minera Llurimagua, la cual está en ejecución. Atelopus
longirostris pertenece a un grupo de no menos de 29 especies de Atelopus de Ecuador que están Críticamente
Amenazadas, 15 de las cuales no han sido vistas en al menos una década y la mayoría de ellas podrían estar extintas. Se
requiere ejecutar más investigaciones simultáneas, multidisciplinarias e integrales para entender la mayoría de aspectos
de la biología de las especies de Atelopus, y las cuales apoyen a los programas de conservación in situ yex situ.
Palabras claves: Atelopus longirostris; Bufonidae; Ecuador; extinción; redescubrimiento
*Corresponding author. Email: firstname.lastname@example.org
© 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unre-
stricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Neotropical Biodiversity, 2017
Vol. 3, No. 1, 157–167, https://doi.org/10.1080/23766808.2017.1327000
Frogs of the genus Atelopus are distributed across tropi-
cal forests, cloud forests and the páramos of Central and
South America. The genus is the largest in the family
Bufonidae, with 96 species described to date  plus
30–70 undescribed . Atelopus has been affected by
catastrophic declines and extinctions; all species
restricted to elevations above 1000 m have declined and
about 75% have disappeared . Atelopus represent
about 15% of the 528 amphibian species that are cur-
rently categorized as Critically Endangered (CR) in the
IUCN Red List of Threatened Species . In Ecuador,
these severe extinction processes have hit a hot spot of
Atelopus diversity given the relatively large number of
species (25 described and at least 7 undescribed, Table 1)
known to occur to date . Twenty-four of the described
species are included in categories of threat in the IUCN
Red List and none in low or no threat categories; among
them 11 are considered Critically Endangered (tagged as
Possibly Extinct ). Thus, the conservation status of
species of Atelopus in Ecuador is of major concern. The
causes of these sudden declines and extinctions of Atelo-
pus species, mostly noticed by the end of the eighties
and ﬁrst half of the nineties, have been a matter of
debate (e.g. [7,8]). Several stressors seem to be the
culprit, most importantly climate change and pathogens
The Longnose harlequin frog, Atelopus longirostris
(Cope 1868), is endemic to the Chocoan region of Ecua-
dor. It used to inhabit the lowlands and subtropics in pre-
montane and montane forests in the Cordillera
Occidental de los Andes of Northwestern Ecuador, at
altitudes between 900 and 1925 m asl. It was last seen in
1989 in Río Esmeraldas (1557 m asl), San Francisco de
Las Pampas, Provincia Cotopaxi, Ecuador. Its historical
records come from 20 localities in the provinces of
Imbabura, Cotopaxi, Pichincha, and Santo Domingo de
los Tsáchilas encompassing an area of extent of occur-
rence (measured by a minimum convex polygon that
contains all the sites of occurrence) of about 1746 Km
in 20 localities from Provincia de Imbabura to Cotopaxi
(Figure 1). The populations of Carchi and Esmeraldas
are excluded because they are in need of a taxonomical
Atelopus longirostris is a diurnal species of terrestrial
and semiarboreal habits. Its activity is associated with
Table 1. Species of Atelopus of Ecuador indicating their political endemicity to Ecuador (E), Red List category (RL), date of most
recent sighting in Ecuador (DS) for possibly extinct species, Ex situ assurance colony (ES).
Species E RL DS ES
Atelopus angelito NE CR 22 July 1988 –
Atelopus arthuri E CR 1 February 1988 –
Atelopus balios E CR Recent Yes*
Atelopus bomolochos E CR Recent Yes
Atelopus boulengeri E CR July 1984 –
Atelopus coynei E CR Recent No
Atelopus elegans NE CR Recent Yes*
Atelopus exiguus E CR Recent Yes
Atelopus guanujo E CR 10 April 1988 –
Atelopus halihelos E CR 28 August 1989 –
Atelopus ignescens E CR Recent Yes
Atelopus longirostris E CR Recent Yes
Atelopus lynchi NE CR 30 May 1977 –
Atelopus mindoensis E CR 7 May 1989 –
Atelopus nanay E CR Recent Yes*
Atelopus nepiozomus E CR Recent Yes
Atelopus onorei E CR 21 April 1990 –
Atelopus orcesi E CR May 1988 –
Atelopus pachydermus NE CR September 1985 –
Atelopus palmatus E CR Recent No
Atelopus pastuso NE CR 29 June 1993 –
Atelopus petersi E CR 8 November 1996 –
Atelopus planispina E CR October 1987 –
Atelopus podocarpus NE CR 1 December 1994 –
Atelopus spumarius NE EN Recent Yes
Atelopus sp. (Limón) NE CR Recent Yes*
Atelopus sp. (Cóndor) E EN Recent Yes
Atelopus sp. (Carchi) E CR Recent No
Atelopus sp. (Azuay) E CR Recent Yes
Atelopus sp. (Sangay) E CR Recent No
Atelopus sp. (Chimborazo) E CR 24 April 2002 –
Atelopus sp. (Pastaza) NE DD Recent No
Notes: An asterisk (*) indicates that breeding has occurred. CR = Critically Endangered, EN = Endangered, DD = Data deﬁcient, Recent = seen at any
date between 2008 and 2016.
158 E.E. Tapia et al.
water streams during the day, where it can be found
walking in opened rocky shores of evergreen forests; by
night it hides under rocks or sleeps on leaves near the
ground. It is a stream breeding species . An
amplexus was reported in 1959 during the end of the
rainy season, and according to the author the female was
heavy with eggs . Besides this, nothing is known
about its biology.
As part of an ongoing inventory of amphibians in the
reserve of the Junín community, Intag, Provincia Imba-
bura, Ecuador, we did extensive searches in the area
from 28 March to 6 April 2016. Among 16 species
found, we report the rediscovery of Atelopus longirostris,
provide additional biological information and discuss
about its conservation.
The study region was located at the reserves of the Junín
community, Cabañas EcoJunín, and also surrounding pri-
vate areas, Cantón Cotacachi, Provincia Imbabura, Ecua-
dor, where we sampled areas between 1159 and 2560 m
asl in foothill, lower montane, and montane cloud forests
from 28 March to 6 April 2016. A second survey
focused on Atelopus longirostris habitat, where we
recorded them previously, was made on 3–4 December
2016. The ﬁrst ﬁeld trip surveys were conducted every
day between 07:00 and 18:00 h during the day, and
between 18:30 and 02:00 h at night, using Visual
Encounter Surveys to record as many amphibians as pos-
sible. Atelopus longirostris total frog search effort was
Figure 1. Localities of ocurrence of Atelopus longirostris in Ecuador. Historical records (black), new record (red).
Neotropical Biodiversity 159
114:30 h. It was done during seven nights (ﬁve nights
for a total of 104:30 h in the ﬁrst survey and two nights
for a total of 10:00 h in the second) time spent in the
potential Atelopus habitat around rivers and streams.
These efforts were divided in the ﬁrst survey as follows:
three nights-three persons (from 18:30 to 02:00 h), one
night-two persons (from 18:30 to 02:00 h), one night-six
persons (from 18:30 to 22:00 h). In the second ﬁeld
trip, search effort was two nights-ﬁve persons (20:00 to
21:00 h). During the ﬁrst survey, tadpole searches were
done during day and night in about two hundred meters
along the river, at the same sites where adults were
found. Clear plastic containers were used as underwater
visors. Also, stones were removed manually to look for
tadpoles at the undersides. Tadpole search effort was
done during two days (from 07:00 to 18:00 h) and one
night (18:30 to 02:00 h) for a total of 29:30 h.
We sampled different types of land cover: forests
(native and secondary), farmlands, grasslands, mixed
areas of agricultural and grasslands, riverbanks (large
rivers and smaller streams), and native bamboo areas.
Information collected in the ﬁeld included: geographic
positions of each encounter, air and water temperature
(°C), time of encounter (24 h), perch height (cm), sex
(when possible), and age class (froglet, juvenile, adult).
Geographic information was recorded using a GPS
GARMIN GPSmap 62s; ph data, water and air tempera-
ture were taken with a HANNA pHep 5 waterproof pH
tester, and with a New RadioShack 22-170 Infrared
Thermometer Pistol Grip Design 10.1 Range. In the
ﬁeld, each individual of Atelopus longirostris was col-
lected and handled with a plastic bag, in which it was
placed. These living individuals were transferred to the
ex-situ conservation program named Life Bank ‘Arca de
los Sapos’of Jambatu Center of Amphibian Research
and Conservation (CJ). Once deposited in the laboratory,
each individual was handled with a fresh pair of latex
gloves to prevent transferring pathogens such as amphib-
ian chytrid fungus (Batrachochytrium dendrobatidis;
Bd), and underwent quarantine. Tests for the presence/ab-
sence of chytrid fungus were done using skin swabs of
Atelopus longirostris and pieces of pelvic patch (stored
in ethanol 75%) of other amphibians. Tests were per-
formed following the standard procedures in Hyatt et al.
; dry swabs were stored in −4 °C until analysis.
DNA from swabs and tissue samples was extracted with
a protocol that uses SDS and Proteinase K for cellular
lysis, guanidine isothiocyanate for protein precipitation
and isopropanol for DNA precipitation. Bd presence was
tested by Polymerase Chain Reaction (PCR) designed to
isolate a 300 bp region of the fungal rDNA using pri-
mers Bd1a (5′-CAGTGTGCCATATGTCACG-3′) and
Bd2a (5′-CATGGTTCATATCTGTCCAG-3′) developed
by Annis et al. . Each PCR reaction contained a ﬁnal
concentration of 3 mM MgCl
, 0.2 mM dNTPs, 0.05 U/μL
Taq DNA polymerase (Invitrogen) and 0.5 μM of each
primer in a 25 μL total volume. PCR protocol followed
Annis et al. , except that 35 cycles were performed.
When the PCR product retrieved was insufﬁcient or
dubious, an additional PCR was carried out, using a 1:50
dilution of the cleaned-up product from the ﬁrst PCR as
template. The conditions of this second PCR were the
same as the ﬁrst one, but fewer cycles were performed.
Two controls: a negative control, containing water
instead of DNA, and a positive control –a sample previ-
ously tested positive for Bd –were used in every PCR.
The presence/absence of Bd was determined via elec-
trophoresis in 1.5% agarose gels. We estimated point
prevalence of Bd within each anuran species as the num-
ber of frogs that tested positive for Bd, divided by the
total number of sampled frogs for that particular species
in our sample.
An ecological characterization of the Reserve of the
Community of Junín is provided by Peñaﬁel Cevallos
et al. . Annual mean temperature varies between
17 and 20 °C, and annual mean humidity varies between
50 and 75%. Mean annual precipitation is 2000–
3000 mm, and the rainy season extends from December
to April whereas the dry season is from May to Novem-
ber. The sampling area (1159–2560 m asl) belongs to the
Foothill Cloud Forest and Montane Cloud Forest in
the subtropical and temperate zoogeographical zones
sensu Albuja et al. . Vegetation at the site is
described by Pazmiño-Otamendi et al. . The lower
parts (about 1159–2000 m asl) are highly disturbed by
human activities, with villages, agricultural areas and
pastures for livestock (Figure 2(A)). In the lower portion,
hilltops are usually deforested because the ﬂat terrain is
optimal for human activity. Despite the impact, there are
some patches of native forest, usually on the slopes of
hills along rivers and streams (Figure 2(B) and (C)). One
of these patches (about 15 hectares) is protected and is
part of Cabañas EcoJunín, which belong to the Junín
community. At higher altitudes, fewer disturbances
occur; however, there are also large deforested frag-
ments, paddocks, and pastures. Areas between 2000 and
2560 m asl belong to the Junín Community Reserve
. These areas are in much better condition, with large
zones of native forest, even on hilltops.
An unexpected ﬁnding in two sites of the lower area of
the sampled region was the presence of 4 adult individu-
als (two males and two females) of Atelopus longirostris.
They were in two small patches of native forest (site 1
of about 21 hectares and site 2 of about 45 hectares, Fig-
ure 2(A) and (B)) associated with the Chalguayacu and
Junín river basins. All the individuals were found at
night between 19:45 h and 22:00 h on 28, 29, and 31
March 2016. Three (one female, two males) of the four
160 E.E. Tapia et al.
individuals were found on site 1, a protected forest of
the reserve of Cabañas EcoJunín, at 1250–1300 m asl.
One female was 15 m from the top of the hill at about
80 m from the Chalguayacu river, whereas the other two
males were 40–50 m from the Chalguayacu river, all of
them on a slope of 60–75%. Another female was found
on site 2 at the top of the hill at about 410 m from the
Chalguayacu river and 200 m from the small river
Argentina (with a river bed about 5 m wide, 50 cm
depth) in a non-protected private property, at 1480 m asl.
The four individuals were found at night resting on
leaves at 40–60 cm above the ground: female CJ (sc
5521) was on a leaf (16 × 6 cm) of Rubiaceae, female
CJ (sc 5583) was on an Anthurium sp. leaf (25 ×
15 cm), male CJ (sc 5522) on a leaf (25 × 20 cm) of
Anthurium sp. growing on a stone (100 × 70 cm) cov-
ered by moss, and male CJ (sc 5582) was on a leaf (9 ×
3.5 cm) of Piper sp. in a bush 2 m high. They were in a
Figure 2. Atelopus longirostris habitat at Junín, Provincia Imbabura: (A–B) Aerial views from an altitude of 11.09 and 2.89 km,
taken from Google, digital Globe; red arrows indicate collection sites at site 1, the reserve of Cabañas EcoJunín (right) and site 2, a
private property (left), scale = 667 and 108 m in A and B, respectively. In ﬁgure B note the nearly complete disconnection between
the private forest and Chalguayacu river caused by forest clearing, (C) forest at reserve of Cabañas EcoJunín, (D) Chalguayacu river,
(E) female CJ (sc 5521) on a leaf of Rubiaceae, (F) male CJ (sc 5582) on a leaf of Piperaceae. Photos C–D by EET, E–F by GPO.
Neotropical Biodiversity 161
patch of secondary forest mixed with fallen trees and
branches. The four frogs were collected, transported to
ex situ breeding facilities of Jambatu Center, and main-
tained in a quarantine period. Two females and one male
survive to date 12 May 2017 in healthy condition. One
male died for undetermined reasons. Latest updates of
their survival status are provided in Centro Jambatu web
The pH of the Chalguayacu river was 7.5 and the
water temperature was 20 °C, taken at 22:00 h on 28
March 2016. Air temperature was 19.4 °C taken at
21:00 h on 29 March 2016; 18 °C taken at 20:30–
22:00 h on 31 March 2016, and 20 °C taken at 07:30 h
on 31 March 2016. The river-bed is about 15 m width,
and water depth can reach about 4 m during the rainy
season, whereas during the dry season stream depth is
about 0.8 m.
Prevalence of Batrachochytrium dendrobatidis in
Pelvic patch (56) and skin swabs (4 of Atelopus lon-
girostris) of 60 frogs of 16 species were tested for Bd,
and a third (20 frogs) of those (belonging to nine
species) were positive for Bd infection (Table 2). The
chytrid analysis of Atelopus longirostris tested negative.
The SVL (snout-vent length) and weight (taken on 28
October 2016) of Atelopus longirostris are as follows:
male CJ (sc 5582) 31.6 mm, 2.2060 g; gravid female CJ
(sc 5521) 36.8 mm, 3.4636 g; female CJ (sc 5583)
39.4 mm, 2.7927 g. External morphology features,
patterns and color in life of male CJ (sc 5582) and
female CJ (sc 5521) are depicted in Figure 3.
During the surveys we recorded observations of 16 spe-
cies of amphibians, seven of which were found at the
same site where we found Atelopus longirostris (Table 2).
Some of them, for example Espadarana prosoblepon
(Boettger 1892), Dendropsophus carnifex (Duellman
1969), Hyloscirtus alytolylax (Duellman 1972), and
Hyloxalus awa (Coloma 1995) occurred at the same col-
lecting site or near Atelopus longirostris microhabitat,
inside the forest associated with rivers or water streams.
The four specimens of Atelopus longirostris have a SVL
within the known range, a swollen gland at tip of snout,
and white pustulae on lateral sides. Thus, they ﬁt well
with previous descriptions of the species [14,22–24].
The species found in sympatry with Atelopus lon-
girostris are nocturnal and mainly arboreal, except for
Hyloxalus awa, which is diurnal, terrestrial, and associ-
ated to streams as Atelopus longirostris , but H. awa
has been found to occupy much more open areas, rather
than native forest areas.
It is interesting that males of Atelopus longirostris
were found relatively far (40–50 m) from the river,
unlike what has been reported in other species of
Atelopus, where some males can be found at the edge of
the breeding sites in both the dry and rainy seasons
[25,26]. If A. longirostris breeds in the Chalguayacu and
or Junín rivers, the absence of males at the river shore
Table 2. Species of amphibians encountered at Cabañas EcoJunín, the Junín Community Reserve, and surrounding areas between
1149 and 2560 m asl, Intag, Provincia Imbabura, Ecuador.
Family Species Endemic RL Bd (n= 60)
Bufonidae Atelopus longirostris* Yes CR –(4)
Centrolenidae Espadarana prosoblepon* No LC + (3/8)
Craugastoridae Pristimantis achatinus* No LC + (4/11)
Craugastoridae Pristimantis appendiculatus No NT –(1)
Craugastoridae Pristimantis dissimulatus Yes EN –(4)
Craugastoridae Pristimantis laticlavius No DD –(1)
Craugastoridae Pristimantis leoni No LC –(3)
Craugastoridae Pristimantis pahuma Yes EN + (1/4)
Craugastoridae Pristimantis pteridophilus Yes E N –(1)
Craugastoridae Pristimantis walkeri* Yes LC + (3/6)
Craugastoridae Pristimantis w-nigrum No EN + (3/5)
Dendrobatidae Hyloxalus awa* Yes VU + (1/4)
Hemiphractidae Gastrotheca plumbea Yes VU Not
Hylidae Dendropsophus carnifex* No LC + (2/3)
Hylidae Hyloscirtus alytolylax* No NT + (1/2)
No LC + (2/3)
Notes: Their political endemicity to Ecuador and Red List category (RL) are indicated. CR = Critically Endangered, EN = Endangered, VU = Vulnerable,
NT = Near Threatened, DD = Data Deﬁcient, LC = Least Concern, n= sample size. An asterisk (*) indicates sympatric species found with Atelopus
longisrostris. Results of Batrachochytrium dendrobatids (Bd) analyses are given as positive or negative (number of positives/total number analyzed).
162 E.E. Tapia et al.
might occur because they avoid large rivers that increase
greatly their water level and current speed during the
Previous to our report, Atelopus longirostris was sighted
27 years ago in May 1989 at San Francisco de Las Pam-
pas, Provincia Cotopaxi. At that time, it was known from
an area of extent of occurrence (measured by a minimum
convex polygon that contains all the sites of occurrence)
of about 1746 Km
in 20 localities from Provincia de
Imbabura to Cotopaxi (Figure 1). Since 1989, San Fran-
cisco de Las Pampas and the protected forest surrounding
it (e.g. Bosque Integral Otonga, BIO) have been the
subject of numerous inventories and studies of ﬂora
and fauna [27,28], including amphibians [29,30], but
A. longirostris has not been found [EET, pers. obs.].
Thus, this population might be extirpated. Search effort at
Las Pampas and BIO have been immense; however, it is
difﬁcult to quantify it given that hundreds of students,
researchers, and reserve guards that have been in the area,
many of them doing amphibian searches during about 3
Additionally, Bustamante et al.  report its
absence in a monitoring study in Río Faisanes, Provincia
Pichincha, a locality where it was recorded until the
mid-eighties. Also no recent records exist from the
Mindo region, Provincia Pichincha, a zone commonly
visited by ecotourists and scientists working on biodiver-
sity . Based on this information, the categorization
of Atelopus longirostris in the IUCN Red List has chan-
ged slightly over time. Chronologically, it has been con-
sidered either Critically Endangered [32,33], Extinct
Figure 3. Live adults of Atelopus longirostris: (A, C, E) CJ (sc 5582), male, SVL = 31.6 mm, (B, D, F) CJ (sc 5521), female,
SVL = 36.8 mm. Photos by LAC.
Neotropical Biodiversity 163
, or Critically Endangered (Possibly Extinct) .
Currently, it ﬁts the Critically Endangered category and
we eliminated the Possibly Extinct tag, which has been
developed to identify those critically endangered species
that are likely already extinct, but for which conﬁrmation
is required .
The western slopes of Cordillera Occidental de los
Andes of Ecuador harbors at least four species of
Atelopus (A. coynei Miyata 1980, A. longirostris,
A. lynchi Cannatella 1981, and A. mindoensis Peters
1973). Among them, a population of A. coynei was
found in 2012 by Andreas Kay , whereas A. lynchi
and A. mindoensis are missing from Ecuador since 1977
and 1989, respectively.
The rediscovery of Atelopus longirostris suggests that
some of the species that were thought as Possibly
Extinct have actually survived strong bottlenecks and are
somehow persisting, even though their populations seem
quite small. In recent years, a relict population of
Rhaebo olallai Hoogmoed 1985 was found , and at
least four species of Ecuadorian Atelopus previously
thought to be Critically Endangered (Possibly Extinct)
have been rediscovered. They are A. nepiozomus Peters
1973 , A. palmatus Andersson 1945 , A. bomolo-
chos Peters 1973 , and A. ignescens (Cornalia 1849)
. These patterns of rediscoveries and persistence and/
or recoveries have been discussed for Atelopus by
Lötters et al. , and are discussed for other taxa in
Central America and North America (e.g. [42,43]).
Whether these patterns of apparent favorable conditions
reported in distant and unrelated places in America
reﬂect common causes or are independent events
requires further investigation. These Atelopus ﬁndings in
Ecuador of either small or seemingly small populations
of Atelopus can be explained by an increase of aware-
ness of the amphibian extinction problem allied to the
increase of batrachologists or naturalists exploring new
areas. Nonetheless, why these populations of a few spe-
cies persisted in only a single or few sites of their once
historical more widespread distribution is a matter of fur-
ther research. Major potential culprits of the sudden
amphibian die-offs and declines such as climate change
and/or pathogens or their interaction [7,11] seem to be,
at least temporally, not acting strongly at sites where
nearly extinct populations have been rediscovered. If
these factors were responsible for the sudden declines,
their persistence suggests that they have changed to mild
conditions. Our ﬁndings of a high point prevalence of
chytrid in the amphibian community at Junín reveals a
similar pattern of Bd prevalence recently described for
Las Gralarias (a locality 33 km South West of the A. lon-
girostris site in Junín) in the western Cordillera de los
Andes . Absence of Bd in A. longirostris might be
explained as a sampling artifact, given the small sample
size (n= 4). At both sites no evidence of mortality due
to Bd was found. In others sites in Ecuador (e.g. Tarvin
et al. ) in the Amazonian region, no evidence of
mortality due to Bd has been found either. Recent data
of the presence of Bd in the Neotropics since historical
times (e.-g. as early as 1863 in the Andes of Bolivia
) challenges previous hypothesis about Bd as the
main culprit of the drastic and enigmatic amphibian
declines especially occurred in the late 1980s and early
1990s; thus further research is needed. Several hypothe-
ses have already been discussed [44–49], among which
interactions between pathogens (e.g. chytrid strains, rana-
virus), host’s evolutionary history, and environmental
factors (e.g. climate change, dry seasons) might be
Nonetheless, it is clear that the genus Atelopus con-
tinues to be highly endangered and that the conservation
of relict populations and species of Atelopus remains a
challenging multidisciplinary task of in situ and ex situ
actions, as has been discussed elsewhere [50,51]. For
example, in Ecuador 29 species of Atelopus are Critically
Endangered, thus nearly extinct, 15 species have not
been sighted in at least 10 years and most of them prob-
ably are extinct, and for none there are genetically viable
populations, although 3 species have been successfully
bred (Table 1).
The reappearance of Atelopus longirostris in the
Intag region of Ecuador constitutes a unique and possi-
bly unrepeatable opportunity to save this endemic spe-
cies from extinction. Pragmatic emergency actions, both
ex situ and in situ, are required to accomplish this objec-
tive [50,52–54]. Atelopus longirostris is a priority species
recommended for ex situ rescue by the Amphibian Con-
servation Needs Assessment workshop for Ecuador, done
in May 21–24 of 2012 . For that purpose, the captive
assurance colony we initiated is a ﬁrst step that would
avoid the impacts of current in situ threats that the spe-
cies currently suffers, such as: chytrid presence and high
prevalence, deforestation, predation, pollution, rising riv-
ers, habitat degradation and fragmentation, trout presence
on the rivers, and mining exploration. Other threats such
as other diseases (e.g. ranaviruses) and climate change
might also be affecting them, but not data at the site are
available. Our initial survey and sampling effort revealed
much fewer individuals than we would expect for a
healthy Atelopus population, thus we suspect that the
population numbers are extremely low and that a bottle-
neck occurred and survival in situ is far from assured.
Certainly the three surviving frogs at the ex situ program
do not grant a genetically viable population either, and
efforts should be taken to increase the number of foun-
ders, especially with a focus on catching tadpoles, bring-
ing them through metamorphosis in laboratory
conditions, and releasing most of them as frogs, when
they might be able to persist better, while some frogs
would be retained as founders. McGregor Reid and
Zippel  summarize and discuss criticism to ex-situ
programs. We have chosen a rapid response , espe-
cially when considering the serious threat of opencast
mining activity that is underway. Also, recent progress
by Centro Jambatu, in developing technologies of main-
tenance and breeding Atelopus in captivity  let us to
164 E.E. Tapia et al.
be optimistic that we are doing the right choice in this
particular case. In contrast, some cases of rediscovered
Atelopus, in low numbers, resulted in the documentation
of their population extirpation or the species possible
Under the assumption that the individuals we found
represent only a portion of a population still existing,
expansion of the reserve of the Junín Community to
include the patch of currently non-protected forest where
Atelopus longirostris occurs is pivotal, as is the restoration
of the habitat between them, to grant the connectivity
among this isolated putative metapopulations. Also, the
restoration of associated river shores is critical to allow the
normal movement of individuals to the breeding sites in
the Chalguayacu and Junín river basins. The fact that we
found females up to 410 m in a straight line from the river,
and males between 40 and 50 m from it, suggests that
females go up to the top of the hills to mature, and then go
back down to the river banks for breeding. For this shift to
occur it is necessary the forest to be in good condition,
from the banks of the river up to the top of the hills.
Current mining activities, which are in the advanced
mining exploration phase, for opencast copper exploita-
tion of the mining concession Llurimagua at the headwa-
ters of the Chalguayacu and Junín rivers are of high
environmental impact . Deforestation to built trails
and well drilling is active at this time. Land slices in the
headwaters of the Chalguayacu and Junín rivers are caus-
ing erosion, resulting in increased sedimentation on the
rocks of these bodies of water. The sedimentation pre-
sumably will affect growth of algae, which are the main
food of Atelopus tadpoles. Additionally, current water
contamination by non-treated thermal waters from well
drilling and other chemicals (e.g. high levels of arsenic as
reported by Knee and Encalada ) are a serious threat
to Atelopus longirostris tadpoles. If mining activity con-
tinues, it will cause serious forms of water pollution .
Because of these factors, it is of great urgency to stop
mining and forest destruction in the area and to prevent
the disposal of any kind of pollutants into the rivers and
streams. Current and potential threats related to mining
activities  should be discouraged. An in situ con-
trolled management program is also essential. For this, it
is a priority to initiate a census and monitoring
program of the species. It is also necessary to further
explore other sites where the species could potentially
exist, especially in Cordillera of Intag and other areas
of its distribution that have not been explored yet.
Simultaneously, it is essential to start studies of the biol-
ogy of the species, with emphasis on its reproductive
biology and behavior. Climate, pathogens, and both phys-
iological and genetic variables associated with the sur-
vival of this population need to be evaluated. This way
we might be able to gain a better understanding of how
this population has survived the environmental and dis-
ease impacts that have been mentioned, whereas other
populations of this and other Atelopus species did not
We are grateful to Carlos Zorrilla, Javier Ramírez and
members of Comunidad de Junín and DECOIN (Defensa
y Conservación Ecológica de Intag), who supported the
inventory work at the Junín region. Javier Ramírez pro-
vided logistic support, housing and hospitality at his home.
Margaux Perchey, Javier Ramírez, Hugo Ramírez,
Oswaldo Ramírez, and Lauro Lucero helped during gen-
eral amphibian ﬁeld collecting. Additionally, Margaux
Perchey enthusiastically helped on exhaustive searches
and the collection of Atelopus longirostris. Diego Acosta-
López diagrammed ﬁgures 2 and 3. Collecting and rearing
of frogs were done under permit 005-15 IC-FAU-DNB/
MA of the Ecuadorian Ministerio de Ambiente (MAE),
issued to Centro Jambatu of Fundación Otonga. Kim Hoke
graciously reviewed a presubmitted version of the manu-
script. Andrew J. Crawford and an anonymous reviewer
provided suggestions that greatly helped to improve our
manuscript. The ex situ management of frogs is supported
by Saint Louis Zoo, Wikiri, and MAE project “Conserva-
tion of Ecuadorian amphibian diversity and sustainable
use of its genetic resources”. We are greatly indebted to
Jeff Bonner, Eric Miller, and Mark Wanner (from Saint
Louis Zoo), and Lola Guarderas (from Wikiri) for their
commitment and sustained support to research and
conservation programs of Ecuadorian threatened frogs.
Associate Editor: W. Chris Funk.
EET and GPO collected specimens, wrote sections of the
MS, and revised the MS. LAC identiﬁed species and wrote
the MS. NP did the chytrid analyses and revised the MS.
No potential conﬂict of interest was reported by the authors.
This study was funded by DECOIN, as well as the MAE pro-
ject ‘Conservation of Ecuadorian amphibian diversity and sus-
tainable use of its genetic resources.’The latter with ﬁnancial
support of the Global Environmental Facility (GEF), and imple-
mentation by Programa de las Naciones Unidas para el Desar-
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