ArticlePDF Available

Abstract and Figures

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 finding 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 individuals 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, multidisciplinary 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.
Content may be subject to copyright.
Full Terms & Conditions of access and use can be found at
http://www.tandfonline.com/action/journalInformation?journalCode=tneo20
Download by: [186.4.158.44] Date: 25 May 2017, At: 07:40
Neotropical Biodiversity
ISSN: (Print) 2376-6808 (Online) Journal homepage: http://www.tandfonline.com/loi/tneo20
Rediscovery of the nearly extinct longnose
harlequin frog Atelopus longirostris (Bufonidae) in
Junín, Imbabura, Ecuador
Elicio Eladio Tapia, Luis Aurelio Coloma , Gustavo Pazmiño-Otamendi &
Nicolás Peñafiel
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:
10.1080/23766808.2017.1327000
To link to this article: http://dx.doi.org/10.1080/23766808.2017.1327000
© 2017 The Author(s). Published by Informa
UK Limited, trading as Taylor & Francis
Group
Published online: 25 May 2017.
Submit your article to this journal
View related articles
View Crossmark data
Rediscovery of the nearly extinct longnose harlequin frog Atelopus longirostris (Bufonidae) in
Junín, Imbabura, Ecuador
Elicio Eladio Tapia
a
, Luis Aurelio Coloma
a,b
*, Gustavo Pazmiño-Otamendi
a
and Nicolás Peñael
c
a
Centro Jambatu de Investigación y Conservación de Anbios, Fundación Otonga, Quito, Ecuador;
b
Universidad Regional
Amazónica IKIAM, Tena, Ecuador;
c
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)
Abstract
We report the recent nding 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, 12501480 m asl, Provincia Imbabura, Ecuador, on 2831 March 2016. The four frogs were found in two isolated
small patches of native forest in a fragmented area heavily modied 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 nd-
ing of A. longirostris after 27 years is surprising and ts 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 rst 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 catastrócos de Atelopus. El redescubrimiento de A.
longisrostris tuvo lugar en una nueva localidad, Junín, 12501480 msnm, Provincia de Imbabura, Ecuador, entre el
2831 de marzo de 2016. Las cuatro ranas se encontraron en dos pequeñas parcelas de bosque natural en un área frag-
mentada y densamente modicada 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 anbios 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 anbios. Estas ranas son
los primeros fundadores de una colonia de manejo ex situ en el Centro Jambatu de Investigación y Conservación de
Anbios. 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: lcoloma@otonga.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, 157167, https://doi.org/10.1080/23766808.2017.1327000
Introduction
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 [1] plus
3070 undescribed [2]. Atelopus has been affected by
catastrophic declines and extinctions; all species
restricted to elevations above 1000 m have declined and
about 75% have disappeared [3]. 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 [4]. 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 [5]. 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 [6]). 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 rst 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
[912].
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
2
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
revision [13].
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 decient, 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 [14]. An
amplexus was reported in 1959 during the end of the
rainy season, and according to the author the female was
heavy with eggs [14]. 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.
Methods
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 34 December
2016. The rst eld 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 (ve nights
for a total of 104:30 h in the rst 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 rst 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 eld
trip, search effort was two nights-ve persons (20:00 to
21:00 h). During the rst 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 eld 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
eld, 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 Saposof 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.
[15]; 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. [16]. Each PCR reaction contained a nal
concentration of 3 mM MgCl
2
, 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. [16], except that 35 cycles were performed.
When the PCR product retrieved was insufcient or
dubious, an additional PCR was carried out, using a 1:50
dilution of the cleaned-up product from the rst PCR as
template. The conditions of this second PCR were the
same as the rst 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.
Results
Study site
An ecological characterization of the Reserve of the
Community of Junín is provided by Peñael Cevallos
et al. [17]. 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 (11592560 m asl) belongs to the
Foothill Cloud Forest and Montane Cloud Forest [18]in
the subtropical and temperate zoogeographical zones
sensu Albuja et al. [19]. Vegetation at the site is
described by Pazmiño-Otamendi et al. [20]. The lower
parts (about 11592000 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 at 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
[17]. These areas are in much better condition, with large
zones of native forest, even on hilltops.
Atelopus longirostris
An unexpected nding 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 12501300 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 4050 m from the Chalguayacu river, all of
them on a slope of 6075%. 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 4060 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: (AB) 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 gure 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 CD by EET, EF 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
page [21].
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
amphibians
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.
Morphology
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.
Sympatric species
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.
Discussion
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 t well
with previous descriptions of the species [14,2224].
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 [14], 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 (4050 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
analyzed
Hylidae Dendropsophus carnifex* No LC + (2/3)
Hylidae Hyloscirtus alytolylax* No NT + (1/2)
Leptodactylidae Leptodactylus
ventrimaculatus*
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 Decient, 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
rainy season.
Conservation
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
2
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 ora
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
difcult 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
decades.
Additionally, Bustamante et al. [31] 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 [13]. 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
[34], or Critically Endangered (Possibly Extinct) [35].
Currently, it ts 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 conrmation
is required [6].
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 [36], 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 [37], 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 [38], A. palmatus Andersson 1945 [38], A. bomolo-
chos Peters 1973 [39], and A. ignescens (Cornalia 1849)
[40]. These patterns of rediscoveries and persistence and/
or recoveries have been discussed for Atelopus by
Lötters et al. [41], 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
reect common causes or are independent events
requires further investigation. These Atelopus ndings 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 ndings 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 [44]. 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. [26]) 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
[45]) 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 [4449], among which
interactions between pathogens (e.g. chytrid strains, rana-
virus), hosts evolutionary history, and environmental
factors (e.g. climate change, dry seasons) might be
involved.
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 [50,5254]. Atelopus longirostris is a priority species
recommended for ex situ rescue by the Amphibian Con-
servation Needs Assessment workshop for Ecuador, done
in May 2124 of 2012 [55]. For that purpose, the captive
assurance colony we initiated is a rst 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 [56] summarize and discuss criticism to ex-situ
programs. We have chosen a rapid response [57], 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 [58] 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
extinction [26,59,60].
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 [59]. 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 [62]) are a serious threat
to Atelopus longirostris tadpoles. If mining activity con-
tinues, it will cause serious forms of water pollution [61].
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 [63] 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
survive.
Acknowledgements
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 eld collecting. Additionally, Margaux
Perchey enthusiastically helped on exhaustive searches
and the collection of Atelopus longirostris. Diego Acosta-
López diagrammed gures 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.
Author contributions
EET and GPO collected specimens, wrote sections of the
MS, and revised the MS. LAC identied species and wrote
the MS. NP did the chytrid analyses and revised the MS.
Disclosure statement
No potential conict of interest was reported by the authors.
Funding
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 nancial
support of the Global Environmental Facility (GEF), and imple-
mentation by Programa de las Naciones Unidas para el Desar-
rollo (PNUD).
ORCID
Luis Aurelio Coloma http://orcid.org/0000-0003-0158-2455
Gustavo Pazmiño-Otamendi http://orcid.org/0000-0002-8788-
7542
References
[1] Frost DR. Amphibian species of the world: an online ref-
erence [Internet]. Version 6.0. New York (NY): American
Museum of National History [cited 2017 Apr 16]. Avail-
able from: http://research.amnh.org/herpetology/amphibia/
index.html
Neotropical Biodiversity 165
[2] Coloma LA, Duellman WE, Almendáriz CA, et al. Five
new (extinct?) species of Atelopus (Anura: Bufonidae)
from Andean Colombia, Ecuador, and Peru. Zootaxa.
2010;54:154.
[3] La Marca E, Lips KR, Lötters S, et al. Catastrophic popu-
lation declines and extinctions in neotropical harlequin
frogs (Bufonidae: Atelopus). Biotropica. 2005;37:190201.
[4] IUCN. The IUCN red list of threatened species [Internet].
Cambridge. 2016 [cited 2017 Apr 16]. Available from:
http://www.iucnredlist.org/
[5] Centro Jambatu. Anbios de Ecuador. SapoPediaEcuador
[Internet]. Fund. Otonga [cited 2017 Apr 13]. Available
from: http://www.anbioswebecuador.ec/index.php?aw,2
[6] IUCN Standards and Petitions Working Group. Guidelines
for using the IUCN Red List categories and criteria. Stan-
dards and Petitions Working Groups of the IUCN SSC
Biodiversity Assesment Sub-Committe; 2008.
[7] Lips KR, Diffendorfer J, Mendelson JR III, et al. Riding
the wave: reconciling the roles of disease and climate
change in amphibian declines. PLoS. 2008;6:441454.
[8] Pounds JA, Coloma LA. Beware the lone killer. Nat Rep
Clim Change. 2008;5759.
[9] Longcore JE, Pessier AP, Nichols DK. Batrachochytrium
Dendrobatidis gen. et sp. nov., a chytrid pathogenic to
amphibians. Mycologia. 1999;91:219227.
[10] Merino-Viteri A, Coloma LA, Almendáriz A. Los Telma-
tobius de los Andes de Ecuador y su disminución pobla-
cional. Monogr Herpetol. 2005;7:937.
[11] Pounds JA, Bustamante MR, Coloma LA, et al.
Widespread amphibian extinctions from epidemic disease
driven by global warming. Nature. 2006;439:161167.
[12] Lips KR, Brem F, Brenes R, et al. Emerging infectious
disease and the loss of biodiversity in a Neotropical
amphibian community. Proc Nat Acad Sci U.S.A.
2006;103:31653170.
[13] Arteaga A, Bustamante L, Guayasamin JM. The amphib-
ians and reptiles of Mindo: life in the cloudforest. Quito:
Scientic Publication Series, Universidad Tecnológica
Indoamérica; 2013.
[14] Peters JA. The frog genus Atelopus in Ecuador (Anura:
Bufonidae). Smithson Contr Zool. 1973;145:149.
[15] Hyatt AD, Boyle DG, Olsen V, et al. Diagnostic assays and
sampling protocols for the detection of Batrachochytrium
dendrobatidis. Dis Aquat Organ. 2007;73:175192.
[16] Annis SL, Dastoor FP, Ziel H, et al. A dna-based assay
identies Batrachochytrium dendrobatidis in amphibians.
J Wildl Dis. 2004;40:420428.
[17] Peñael Cevallos M, Tirado M, Castro D, et al. Estudio
de caracterización ecológica de la reserva comunitaria de
Junín. Apuela: DECOIN (Defensa y Conservación Ecoló-
gica de Intag), Alianza Jatun Sacha-Centro de Datos para
la Conservación; 2005.
[18] Ministerio de Ambiente del Ecuador. Sistema de clasi-
cación de los ecosistemas del Ecuador continental. Sub-
secretaría de Patrimonio Natural (a) [Internet]. Quito;
2012. Available from: http://www.ambiente.gob.ec/wp-con
tent/uploads/downloads/2012/09/LEYENDA-ECOSISTE
MAS_ECUADOR_2.pdf
[19] Albuja L, Almendáriz A, Barriga R, et al. Fauna de verte-
brados del Ecuador. Quito: Instituto de Ciencias Biológi-
cas. Escuela Politécnica Nacional; 2012.
[20] Pazmiño-Otamendi G, Tapia EE, Coloma LA. Guía de
anbios de la comunidad Junín, Intag, Provincia de Imba-
bura, Ecuador. Quito: Centro Jambatu, DECOIN (Defensa
y Conservación Ecológica de Intag); 2016.
[21] Centro Jambatu. Saparium. Anbios vivos conservación
[Internet]. Fund. Otonga; 2017. Available from: http://
www.anbioswebecuador.ec/index.php?as,17
[22] Cannatella DC. A new Atelopus from Ecuador and
Colombia. J Herpetol. 1981;15:133138.
[23] Lötters S. The Neotropical toad genus Atelopus. Checklist
biology distribution. Köln: M. Vences & F. Glaw; 1996.
[24] Coloma LA. Morphology, systematics and phylogenetic
relationships amog frogs of the genus Atelopus (Anura:
Bufonidae) [dissertation]. Lawrence (KS): University of
Kansas; 1997.
[25] Lampo M, Señaris JC, Rodríguez-Contreras A, et al. High
turnover rates in remnant populations of the harlequin frog
Atelopus cruciger (Bufonidae): low risk of extinction?
Biotropica. 2012;44:420426.
[26] Tarvin RD, Peña P, Ron SR. Changes in population size
and survival in Atelopus spumarius (Anura: Bufonidae)
are not correlated with chytrid prevalence. J Herpetol.
2014;48:291297.
[27] Dupérré N, Tapia E. Overview of the Anyphaenids (Ara-
neae, Anyphaeninae, Anyphaenidae) spider fauna from the
Chocó forest of Ecuador, with the description of thirteen
new species. Zootaxa. 2016;255:150.
[28] Kizirian DA. A review of Ecuadorian Proctoporus (Squa-
mata: Gymnophthalmidae) with descriptions of nine new
species. Herpetol Monogr. 1996;10:85155.
[29] Guayasamin JM, Bonaccorso E, Menéndez PA, et al. Mor-
phological variation, diet, and vocalization of Eleuthero-
dactylus eugeniae (Anura: Leptodactylidae) with notes on
its reproduction and ecology. Herpetol Rev. 2004;35:1723.
[30] Guayasamin JM, Hutter CR, Tapia EE, et al. Diversica-
tion of the rainfrog Pristimantis ornatissimus in the low-
lands and Andean foothills of Ecuador. PLOS One.
2017;12:e0172615.
[31] Bustamante MR, Ron SR, Coloma LA. Changes in diver-
sity of seven anuran communities in the Ecuadorian
Andes. Biotropica. 2005;37:180189.
[32] Coloma LA. Anbios de Ecuador: estatus poblacional y
de conservación. Quito: Centro de Datos para la Conser-
vación-Ecuador; 1992.
[33] Ron SR, Guayasamin JM, Coloma LA, et al. Lista roja de
los anbios de Ecuador [Internet]. Version 1.0 (2 de mayo
2008). Mus. Zool. Pontif. Univ. Católica del Ecuador;
2008 [cited 2017 Apr 13]. Available from: https://web.ar
chive.org/web/20090917184123/http://www.puce.edu.ec/zo
ologia/sron/roja%0D405/index.htm
[34] Bustamante MR, Bolívar W, Coloma LA, et al. Atelopus
longirostris [Internet]. IUCN Red List Threat. Species.
Version 2015-4. 2004 [cited 2017 Apr 13]. Available
from: http://dx.doi.org/10.2305/IUCN.UK.2004.RLTS.T5
4522A11158637.en
[35] Coloma LA, Guayasamin JM, Menéndez-Guerrero P
(eds). Lista roja de anbios de Ecuador. SapoPediaEcua-
dor [Internet]. Centro Jambatu; 2017. Available from:
http://www.anbioswebecuador.ec/index.php?lr,10
[36] Kay A. Atelopus coynei observed February 7, 2012,
research-grade observations [Internet]. Ina. San Fr. USA
Calif. Acad. Sci; 2012 [cited 2017 Apr 13]. Available
from: http://www.inaturalist.org/observations/55996
[37] Lynch RL, Kohn S, Ayala-Varela F, et al. Rediscovery of
Andinophryne olallai Hoogmoed, 1985 (Anura, Bufoni-
dae), an enigmatic and endangered Andean toad. Amphib
Reptil Conserv. 2014;8:17.
[38] Yánez-Muñoz MH, Veintimilla DA, Smith EN, et al. Des-
cubrimiento de dos poblaciones sobrevivientes de sapos
arlequín (Amphibia: Bufonidae: Atelopus) en los Andes de
Ecuador. 2010;2:25.
[39] Holland J. Extincttoad rediscovered in Ecuador [Internet].
Natl. Geogr. Weird Wild. 2015 [cited 2017 Apr 13]. Available
from: http://news.nationalgeographic.com/2015/08/150831-
frogs-extinct-rediscovered-science-animals-ecuador/
166 E.E. Tapia et al.
[40] Coloma LA. El Jambato negro del páramo, Atelopus
ignescens, resucitó [Internet]. IMCiencia. 2016. Available
from: http://www.imciencia.com/el-jambato-negro-del-
paramo-atelopus-ignescens-resucito/
[41] Lötters S, La Marca E, Gagliardo RW, et al. Harlequin
frogs back. Some thoughts and speculations. Froglog.
2005;70:13.
[42] Abarca J, Chaves G, García-Rodríguez A, et al. Reconsid-
ering extinction: rediscovery of Incilius holdridgei (Anura:
Bufonidae) in Costa Rica after 25 years. Herpetol Rev.
2010;41:150.
[43] Knapp RA, Fellers G, Kleeman P, et al. Large-scale recov-
ery of an endangered amphibian despite ongoing exposure
to multiple stressors. Proc Nat Acad Sci. 2016;113:
1188911894.
[44] Guayasamin JM, Mendoza ÁM, Longo AV, et al. High
prevalence of Batrachochytrium dendrobatidis in an
Andean frog community (Reserva Las Gralarias, Ecuador).
Amphib Reptil Conserv Amphib Reptil Conserv.
2014;8:3344.
[45] Burrowes PA, De la Riva I. Unraveling the historical
prevalence of the invasive chytrid fungus in the Bolivian
Andes: implications in recent amphibian declines. Biol.
Invasions. 2017:114.
[46] Pounds JA, Crump ML. Amphibian declines and climate
disturbance the case of the golden toad and the harle-
quin frog. Conserv Biol. 1994;8:7285.
[47] Whiteld SM, Geerdes E, Chacon I, et al. Infection and
co-infection by the amphibian chytrid fungus and rana-
virus in wild Costa Rican frogs. Dis Aquat Organ.
2013;104:173178.
[48] Warne RW, La Bumbard B, La Grange S, et al. Co-infection
by chytrid fungus and ranaviruses in wild and harvested
frogs in the tropical Andes. PLoS One. 2016;11:115.
[49] Lampo M, Rodríguez-Contreras A, La Marca E, et al. A
chytridiomycosis epidemic and a severe dry season pre-
cede the disappearance of Atelopus species from the
Venezuelan Andes. Herpetol J. 2006;16:395402.
[50] Lötters S. The fate of the harlequin toads help through a
synchronous multidisciplinary approach and the IUCN
Amphibian Conservation Action Plan. Mitteilungen
Museum Für Naturkd Berlin Zool R. 2007;83:6973.
[51] Stuart SN. Responding to the amphibian crisis: too little,
too late. Alytes. 2012;29:912.
[52] Mendelson JR III, Lips KR, Gagliardo RW, et al. Con-
fronting amphibian declines and extinctions. Science.
2006;313:4848.
[53] Scheele BC, Hunter DA, Grogan LF, et al. Interventions
for reducing extinction risk in chytridiomycosis-threatened
amphibians. Conserv Biol. 2014;28:11951205.
[54] Zippel KC, Mendelson JR III. The amphibian extincion
crisis: a call to action. Herpetol Rev. 2008;39:2329.
[55] AARK. Amphibian conservation needs assessment work-
shop for Ecuador [Internet]. 2012 [cited 2012 May 21
24]. Available from: http://www.conservationneeds.org/
SpeciesRecommendRescue.aspx
[56] McGregor Reid G, Zippel KC. Can zoos and aquariums
ensure the survival of amphibians in the 21st century? Int
Zoo Yearb. 2008;42:16.
[57] Gagliardo R, Crump P, Grifth E, et al. The principles of
rapid response for amphibian conservation, using the pro-
grammes in Panama as an example. Int Zoo Yearb.
2008;42:125135.
[58] Coloma LA, Almeida-Reinoso DP. Ex situ management of
ve extant species of Atelopus in Ecuador. Progress
report. AARK Newsl. 2012;20:812.
[59] Salazar-Valenzuela D. Demografía e historia natural de
una de las últimas ranas arlequín (Atelopus sp.) (Anura:
Bufonidae) del Ecuador [Licenciatura thesis]. Quito: Pon-
ticia Universidad Católica del Ecuador; 2007.
[60] Lampo M, Barrio-Amorós C, Han B. Batrachochytrium den-
drobatidis infection in the recently rediscovered Atelopus
mucubajiensis (Anura, Bufonidae), a critically endangered frog
from the Venezuelan Andes. Ecohealth. 2006;3:299302.
[61] Chopard A, Sacher W. Megaminería y agua en Íntag: una
evaluación independiente. Análisis preliminar de los
potenciales impactos en el agua por la explotación de
cobre a cielo abierto en Junín, zona de Intag, Ecuador.
Apuela: DECOIN (Defensa y Conservación Ecológica de
Intag); 2017.
[62] Knee K, Encalada A. La calidad del agua en la zona de
Íntag (Imbabura) y su relación con el uso del suelo.
Apuela: DECOIN (Defensa y Conservación Ecológica de
Intag); 2012.
[63] Kocian M, Batker D, Harrison-Cox J. An ecological study
of Ecuadors Intag region: The environmental impacts and
potential rewards of mining. Tacoma, WA: Earth Eco-
nomics; 2011.
Neotropical Biodiversity 167
... Atelopus is the largest genus of the bufonids with 96 species described to date [1], and many others awaiting description (e.g. Tapia et al. [2]: Table 1). They are distributed in the tropical rainy and cloud forests, and along the paramos of Central and South America [2]. ...
... Tapia et al. [2]: Table 1). They are distributed in the tropical rainy and cloud forests, and along the paramos of Central and South America [2]. More than 90% of the species is either Endangered, Critically Endangered and/or Possibly extinct, and the rest are either Data Deficient, Non-threatened or not evaluated [2][3][4]. ...
... They are distributed in the tropical rainy and cloud forests, and along the paramos of Central and South America [2]. More than 90% of the species is either Endangered, Critically Endangered and/or Possibly extinct, and the rest are either Data Deficient, Non-threatened or not evaluated [2][3][4]. Their tadpoles belong to the gastromyzophorus ecomorphological guild, which means that they use an abdominal sucker to keep their position in fast and turbulent streams and rivers [5,6]. However, there are some species from the lowlands that also exploit slow running streams [7]. ...
Article
Full-text available
The tadpoles of the Neotropical genus Atelopus are only known for 26 out of 96 species described. Here, we describe the tadpoles of A. elegans and A. palmatus including ontogenetic information, measurements, and images of individuals in several stages of growth. Both species are compared with their congeners taking into account some relevant features such as the coloration and relative measurements. Our description focuses on the abdominal sucker and mouth by providing scanning electron microscopy images and comparing the suctorial mechanism with other groups of anurans and fish. We also provide an update to knowledge of the abdominal suckers, and information about their lateral line system and the distribution of their lateral line openings. The results show that brown marks over a tan surface and an irregular distribution of marks along the body and tail are unique to A. elegans; while a patterned distribution of contrasting marks, and the presence of submarginal papillae are unique to A. palmatus. Also, both species show differences in the structures of their teeth. Finally, we conclude that some characters such as coloration, presence or absence of some structures, and relative measurements are useful for identifying the species.
... Lampo et al. (2017) suggest that this lowland species contradicts thermal refuge hypothesis patterns (i.e., that warmer lowland regions serve as refugia from Bd, which prefers cooler temperatures; Piotrowski et al., 2004;Woodhams et al., 2008), and may be able to persist because of decreased disease transmission. Most other studies (including in Central America) are short reports that have arisen in the past decade documenting the presence of Bd or first sighting of long-lost species or populations (e.g., González-Maya et al., 2013;Tapia et al., 2017;Jiménez-Monge et al., 2019;Barrio Amorós et al., 2020). An analysis focused on the 32 putative species of Atelopus found in Ecuador suggested that more than half of the species not seen after abrupt declines were found in recent years (Tapia et al., 2017). ...
... Most other studies (including in Central America) are short reports that have arisen in the past decade documenting the presence of Bd or first sighting of long-lost species or populations (e.g., González-Maya et al., 2013;Tapia et al., 2017;Jiménez-Monge et al., 2019;Barrio Amorós et al., 2020). An analysis focused on the 32 putative species of Atelopus found in Ecuador suggested that more than half of the species not seen after abrupt declines were found in recent years (Tapia et al., 2017). While the documentation of rediscovered species has developed greatly in the past decade, further investigation is needed across species, environments, and decline histories to gain a full understanding of mechanisms of persistence in Atelopus, and to apply this understanding more broadly to other amphibians. ...
Article
Full-text available
Amphibians face global declines, and it remains unclear the extent to which species have responded, and through what mechanisms, to persist in the face of emerging diseases and climate change. In recent years, the rediscovery of species considered possibly extinct has sparked public and scientific attention. These are hopeful cases in an otherwise bleak story. Yet, we know little about the population status of these rediscovered species, or the biology underlying their persistence. Here, we highlight the iconic Harlequin frogs (Bufonidae: Atelopus) as a system that was devastated by declines but now encompasses between 18 and 32 rediscoveries (25–37 % of possible extinctions) in the last two decades. Geographic distributions of rediscoveries closely match regional described species abundance, and rediscoveries are documented at elevations from 100 m to >3500 m, with no significant differences between mean historical and contemporary elevations. We also provide genomic data on six decimated species, with historical sample comparisons for two of the species and find a pattern of decreasing genetic variation the longer a species had been missing. Further, we document marked decrease in heterozygosity in one species, but not the other, indicating potential idiosyncratic consequences of declines. Finally, we discuss research priorities to guide the potential transition from amphibian declines to recoveries and to maximize conservation efforts.
... Chytridiomycosis, a disease caused by the fungal pathogen Batrachochytrium dendrobatidis, is implicated in many of the declines (La Lampo et al., 2017), and habitat loss and degradation are likely also important drivers (Gómez-Hoyos et al., 2020;Jorge et al., 2020b;Santa-Cruz et al., 2017). Recently, several Atelopus species thought to be extinct or locally extirpated have been rediscovered (Barrio Amorós et al., 2020;Enciso-Calle et al., 2017;Escobedo-Galván et al., 2013;Tapia et al., 2017); however, these rediscovered populations are still at risk of extinction due to habitat loss, invasive species, low genetic diversity, and chytridiomycosis (Byrne et al., 2020;González-Maya et al., 2018;Kardos et al., 2021). Atelopus extinctions not only risk the loss of irreplaceable biodiversity but also threaten the persistence of toxins that are unique to the genus. ...
... Amazonian and Central Andean species have received particularly little investigation. Although Ecuador is a center of Atelopus diversity (25 described species, of which 17 are endemic; Tapia et al., 2017), populations from only two Atelopus species (A. planispina and A. "ignescens") in Ecuador have been assessed (Table S2). ...
Article
Full-text available
Toads of the genus Atelopus are chemically defended by a unique combination of endogenously synthesizedcardiotoxins (bufadienolides) and neurotoxins which may be sequestered (guanidinium alkaloids). Investigationinto Atelopus small-molecule chemical defenses has been primarily concerned with identifying and characterizingvarious forms of these toxins while largely overlooking their ecological roles and evolutionary implications. Inaddition to describing the extent of knowledge about Atelopus toxin structures, pharmacology, and biologicalsources, we review the detection, identification, and quantification methods used in studies of Atelopus toxins todate and conclude that many known toxin profiles are unlikely to be comprehensive because of methodologicaland sampling limitations. Patterns in existing data suggest that both environmental (toxin availability) andgenetic (capacity to synthesize or sequester toxins) factors influence toxin profiles. From an ecological andevolutionary perspective, we summarize the possible selective pressures acting on Atelopus toxicity and toxinprofiles, including predation, intraspecies communication, disease, and reproductive status. Ultimately, weintend to provide a basis for future ecological, evolutionary, and biochemical research on Atelopus.
... More than 90% of a hundred and plus species of harlequin frogs of the genus Atelopus are possibly extinct, critically endangered or endangered under IUCN criteria [33,60]. Among them is our studied species Atelopus sp. ...
... (spumarius complex, A. sp 13 sensu Arbeláez-Ortiz [1]). This species is considered critically endangered, although its formal description has not been published yet [60]. Successful attempts to maintain and breed this species under laboratory conditions have been reported, the first one occurred at Centro Jambatu de Investigación y Conservación de Anfibios (CJ) in 2011 [21] and latter since November 2014 (CJ laboratory notes by Patricio Vargas-Mena). ...
Article
Amphibians are in peril, given the ongoing sixth mass extinction of wildlife. Thus, Conservation Breeding Programs (CBPs) are attempting to breed some species under laboratory conditions. The incorporation of assisted reproduction technologies (ARTs), such as hormonal stimulation, sperm collection and cryopreservation, and in vitro fertilization is contributing to successful CBPs. The objective of this study was to apply ARTs in sexually mature individuals of an undescribed species of Atelopus (spumarius complex) (harlequin frog). Our procedure involves hormonal induction of gametogenesis in this species. We were able to induce gamete release through administration of human chorionic gonadotropin (hCG) in males, and in females this has been achieved through the sequential administration of hCG (priming doses), and combinations of hCG with gonadotropin releasing hormone analogue, GnRHa (ovulary dose). We standardized sperm cryopreservation by performing toxicity tests of cryoprotectants, fast/slow freezing and thawing, as well as supplementation of non-penetrating cryoprotectants (sugars). Next, we performed in vitro fertilization, evaluated the fertilization capacity of the cryopreserved sperm, and describe external features of fresh and cryopreserved sperm. We found that 10 IU/g hCG induced the release of the highest sperm concentrations between 3 and 5 h post-injection, while 2.5 IU/g hCG induced the release of eggs in most treated females. Under cryopreservation conditions, the highest recovery of forward progressive motility or FPM was 26.3 ± 3.5%, which was obtained in cryosuspensions prepared with the 5% DMF and 2.5% sucrose. Cryopreserved sperm showed narrower mitochondrial vesicles after thawing, while in frozen samples without cryodiluent showed 31% of spermatozoa lost their tails. In most cases, our attempts of in vitro fertilization were successful. However, only ∼10% of embryos were viable. Overall, our study demonstrates that the development of ARTs in individuals of Atelopus sp. (spumarius complex) bred in laboratory can be successful, which result in viable offspring through in vitro fertilization. Our study provides a baseline for assisted breeding protocols applicable to other harlequin frogs of the genus Atelopus.
... Some species of harlequin toads have been rediscovered in recent years (A. varius [13], A. nepiozomus [14], A. palmatus [14], A. bomolochos [15], A. ignescens [16], A. nanay [17], A. longirostris [18] and A. carrikeri [19]), and few populations appear to be recovering despite the presence of Bd in some of its individuals [20][21][22][23]. The mechanisms that allow these populations to coexist with the fungus are not fully understood. ...
Article
Full-text available
Chytridiomycosis, a disease caused by the fungus Batrachochytrium dendrobatidis ( Bd ), has been linked with the disappearance of amphibian populations worldwide. Harlequin toads ( Atelopus ) are among the most severely impacted genera. Two species are already considered extinct and most of the others are at high risk of extinction. The recent rediscovery of harlequin toad populations coexisting with Bd suggest that the pathogen can maintain enzootic cycles at some locations. The mechanisms promoting coexistence, however, are not well understood. We explore the dynamics of Bd infection in harlequin toads by modeling a two-stage host population with transmission through environmental reservoirs. Simulations showed that variations in the recruitment of adults and the persistence of zoospores in the environment were more likely to drive shifts between extinction and coexistence than changes in the vulnerability of toads to infection with Bd . These findings highlight the need to identify mechanisms for assuring adult recruitment or minimizing transmission from potential reservoirs, biotic or abiotic, in recovering populations.
... On the other hand, a species is considered tolerant if it has the capacity to limit the consequences of the infection; in this case, although the pathogen is present, the animals do not die from chytridiomycosis [7]. Although the decline of amphibians observed in the 1980s and 1990s led to their disappearance, several species of amphibians have been rediscovered since the year 2000 [31][32][33][34][35][36][37][38][39]. It remains unclear why certain populations or species of amphibians survive whereas others are extirpated with the appearance of Bd. ...
Article
Full-text available
Amphibians have declined around the world in recent years, in parallel with the emergence of an epidermal disease called chytridiomycosis, caused by the chytrid fungus Batrachochytrium dendrobatidis ( Bd ). This disease has been associated with mass mortality in amphibians worldwide, including in Costa Rica, and Bd is considered an important contributor to the disappearance of this group of vertebrates. While many species are susceptible to the disease, others show tolerance and manage to survive infection with the pathogen. We evaluated the pathogen Bd circulating in Costa Rica and the capacity of amphibian skin bacteria to inhibit the growth of the pathogen in vitro . We isolated and characterized – genetically and morphologically – several Bd isolates from areas with declining populations of amphibians. We determined that the circulating chytrid fungus in Costa Rica belongs to the virulent strain Bd -GPL-2, which has been related to massive amphibian deaths worldwide; however, the isolates obtained showed genetic and morphological variation. Furthermore, we isolated epidermal bacteria from 12 amphibian species of surviving populations, some in danger of extinction, and evaluated their inhibitory activity against the collection of chytrid isolates. Through bioassays we confirmed the presence of chytrid-inhibitory bacterial genera in Costa Rican amphibians. However, we observed that the inhibition varied between different isolates of the same bacterial genus, and each bacterial isolation inhibited fungal isolation differently. In total, 14 bacterial isolates belonging to the genera Stenotrophomonas , Streptomyces , Enterobacter , Pseudomonas and Klebsiella showed inhibitory activity against all Bd isolates. Given the observed variation both in the pathogen and in the bacterial inhibition capacity, it is highly relevant to include local isolates and to consider the origin of the microorganisms when performing in vivo infection tests aimed at developing and implementing mitigation strategies for chytridiomycosis.
... In Ecuador, evidence of the scale of the declines in individual species has been difficult to quantify, and there have been few systematic B. dendrobatidis surveys at a landscape scale (Bresciano et al., 2015). However, published field surveys have showed that once-common species such as the Quito stubfoot toad (Atelopus ignescens) experienced total extirpation across known populations (Ron et al., 2003) while recent efforts revealed that some species have been persisting in isolated relict populations, including the Quito stubfoot toad and other Atelopus species Tapia et al., 2017). ...
Article
Full-text available
Microbiome‐pathogen interactions are increasingly recognised as an important element of host immunity. While these host‐level interactions will have consequences for community disease dynamics, the factors which influence host microbiomes at larger scales are poorly understood. We here describe landscape scale pathogen‐microbiome associations within the context of post‐epizootic amphibian chytridiomycosis, a disease caused by the panzootic chytrid fungus Batrachochytrium dendrobatidis. We undertook a survey of Neotropical amphibians across altitudinal gradients in Ecuador ~30 years following the observed amphibian declines and collected skin swab‐samples which were metabarcoded using both fungal (ITS‐2) and bacterial (r16S) amplicons. Our data reveals marked variation in patterns of both B. dendrobatidis infection and microbiome structure that are associated with host life history. Stream breeding amphibians were most likely to be infected with B. dendrobatidis. This increased probability of infection was further associated with increased abundance and diversity of non‐Batrachochytrium chytrid fungi in the skin and environmental microbiome. We also show that increased alpha diversity and the relative abundance of fungi is lower in the skin microbiome of adult stream amphibians compared to adult pond breeding amphibians, an association not seen for bacteria. Finally, stream tadpoles exhibit lower proportions of predicted protective microbial taxa than pond tadpoles, suggestive of reduced biotic resistance. Our analyses show that host breeding ecology strongly shapes pathogen‐microbiome associations at a landscape scale, a trait that may influence resilience in the face of emerging infectious diseases.
... liver and muscle), and provide troubleshooting tips to deal with different experimental situations. Using this protocol, we have successfully retrieved DNA from muscle, skin, and liver of vertebrates for downstream PCR applications in phylogenetics [4], phylogeography [5], and diversification [6], but we have also adapted the protocol to isolate DNA from avian blood samples kept in 99% ethanol for malaria detection [7] and from skin swabs of amphibians to detect Batrachochytrium dendrobatidis (Bd) infection [8]. ...
Article
Full-text available
Laboratories that perform PCR on a routine basis need to count on reliable DNA isolation methods. In locations where supply of DNA extraction kits depends on importation, having an in-house protocol is desirable. This is also important for laboratories limited by budget constraints. We present a low-cost DNA isolation protocol that incorporates well-known techniques, but that we have adapted to various animal tissues. We tested this protocol on animal blood and muscle, and on cell suspension from skin swabs. The results were comparable, in terms of amount and quality of DNA, to those obtained with two other commercially available methods. DNA retrieved with this protocol has been successfully employed for Sanger sequencing of gene PCR products from animal tissues and blood, as well as for PCR-based diagnosis of chytrid fungus in amphibians and blood parasites in birds.
Article
Full-text available
Amphibian chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd), has been recognized as the infectious disease causing the most catastrophic loss of biodiversity known to science, with South America being the most impacted region. We tested whether Bd prevalence is distributed among host taxonomy, ecoregion, conservation status and habitat preference in South America. Here we provide a synthesis on the extent of Bd infection across South America based on 21 648 molecular diagnostic assays, roles of certain species in the epidemiology of Bd and explore its association with the reported amphibian catastrophic declines in the region. We show that Bd is widespread, with a continental prevalence of 23.2%. Its occurrence in the region shows a phylogenetic signal and the probability of infection is determined by ecoregion, preferred habitat and extinction risk hosts' traits. The taxa exhibiting highest Bd occurrence were mostly aquatic amphibians, including Ranidae, Telmatobiidae, Hylodidae, Calyptocephalellidae and Pipidae. Surprisingly, families exhibiting unusually low Bd prevalence included species in which lethal chytridiomycosis and population declines have been described (genera Atelopus, Rhinoderma and Eleutherodactylus). Higher than expected prevalence of Bd occurred mainly in amphibians living in association with mountain environments in the Andes and Atlantic forests, reflecting highly favourable Bd habitats in these areas. Invasive amphibian species (e.g. Lithobates catesbeianus and Xenopus laevis) exhibited high Bd prevalence; thus we suggest using these as sentinels to understand their potential role as reservoirs, vectors or spreaders of Bd that can be subjected to management. Our results guide on the prioritization of conservation actions to prevent further biodiversity loss due to chytridiomycosis in the world's most amphibian diverse region.
Article
Avoiding extinction in a rapidly changing environment often relies on a species' ability to quickly adapt in the face of extreme selective pressures. In Panamá, two closely related harlequin frog species (Atelopus varius and Atelopus zeteki) are threatened with extinction due to the fungal pathogen Batrachochytrium dendrobatidis (Bd). Once thought to be nearly extirpated from Panamá, A. varius have recently been rediscovered in multiple localities across their historical range; however, A. zeteki are possibly extinct in the wild. By leveraging a unique collection of 186 Atelopus tissue samples collected before and after the Bd outbreak in Panama, we describe the genetics of persistence for these species on the brink of extinction. We sequenced the transcriptome and developed an exome-capture assay to sequence the coding regions of the Atelopus genome. Using these genetic data, we evaluate the population genetic structure of historical A. varius and A. zeteki populations, describe changes in genetic diversity over time, assess the relationship between contemporary and historical individuals, and test the hypothesis that some A. varius populations have rapidly evolved to resist or tolerate Bd infection. We found a significant decrease in genetic diversity in contemporary (compared to historical) A. varius populations. We did not find strong evidence of directional allele frequency change or selection for Bd resistance genes, but we uncovered a set of candidate genes that warrant further study. Additionally, we found preliminary evidence of recent migration and gene flow in one of the largest persisting A. varius populations in Panamá, suggesting the potential for genetic rescue in this system. Finally, we propose that previous conservation units should be modified, as clear genetic breaks do not exist beyond the local population level. Our data lay the groundwork for genetically informed conservation and advance our understanding of how imperiled species might be rescued from extinction.
Article
Full-text available
We studied populations of frogs of the genus Atelopus from the Pasto Massif of the Andes in southern Colombia and northern Ecuador, and from the Huancabamba depression in southern Ecuador and northern Perú and conclude that they belong to six species, five of which are described as new to science. Atelopus angelito is recorded for the first time from Ecuador and its range is extended 183 km (airline) southwest of its type locality in Departamento del Cauca, Colombia. We distinguish the five new species from similar ones using features of coloration, skin texture, and morphometrics. We also include osteological data for four of the new species. A putative hybrid zone at Provincia Imbabura, Ecuador, is proposed to exist between the non-sister taxa A. ignescens and one of the new species. Because recent records of four of the new species and A. angelito are lacking despite search efforts, we hypothesize that they are possibly extinct, as are many other Andean Atelopus. Thus, we categorize these species, applying IUCN Red List criteria, as Critically Endangered (Possibly Extinct). No search efforts have been carried out for one new species (from La Victoria, Colombia); thus, it is included under the Data Deficient category. The conservation of Atelopus is briefly discussed.
Technical Report
Full-text available
En el presente estudio, ofrecemos un análisis riguroso e independiente del potencial contaminante del agua, que se produciría por la eventual explotación de cobre a cielo abierto en Junín, zona de Íntag, Ecuador, por parte de las empresas Enami de Ecuador y Codelco de Chile. A partir de muestras recopiladas en el sitio de la mina proyectada, establecemos la presencia de una serie de elementos químicos y minerales en las rocas del sitio, conocidos por generar graves proble- mas, como la contaminación del agua con Drenaje Ácido de Mina, metales pesados y otros elementos tóxicos. Calculamos además que la cantidad de desechos mineros de una mina a cielo abierto en Junín alcanzaría al menos varios cientos de millones de toneladas. Para terminar, mostramos que otras minas a cielo abierto, en contextos similares al de Junín, han provocado graves formas de con- taminación del agua. Estas características, añadidas al contexto sensible del territorio del proyecto, tanto física, como am- biental, económica y socialmente, nos llevan a formular una serie de recomendaciones específicas para las autoridades ecuatorianas. Además, nos parece que constituyen un conjunto de elementos que cuestionan profundamente la minería a gran escala en Junín y la zona de Intag en su conjunto. Palabras claves: Minería a gran escala, Ecuador, previsión de impactos, potencial contaminante del agua, potencial de generación de ácido, espectrometría, difracción de rayos X, microscopía óptica y electrónica
Article
Full-text available
Geographic barriers and elevational gradients have long been recognized as important in species diversification. Here, we illustrate an example where both mechanisms have shaped the genetic structure of the Neotropical rainfrog, Pristimantis ornatissimus, which has also resulted in speciation. This species was thought to be a single evolutionary lineage distributed throughout the Ecuadorian Chocó and the adjacent foothills of the Andes. Based on recent sampling of P. ornatissimus sensu lato, we provide molecular and morphological evidence that support the validity of a new species, which we name Pristimantis ecuadorensis sp. nov. The sister species are elevational replacements of each other; the distribution of Pristimantis ornatissimus sensu stricto is limited to the Ecuadorian Chocó ecoregion (< 1100 m), whereas the new species has only been found at Andean localities between 1450–1480 m. Given the results of the Multiple Matrix Regression with Randomization analysis, the genetic difference between P. ecuadorensis and P. ornatissimus is not explained by geographic distance nor environment, although environmental variables at a finer scale need to be tested. Therefore this speciation event might be the byproduct of stochastic historic extinction of connected populations or biogeographic events caused by barriers to dispersal such as rivers. Within P. ornatissimus sensu stricto, morphological patterns and genetic structure seem to be related to geographic isolation (e.g., rivers). Finally, we provide an updated phylogeny for the genus, including the new species, as well as other Ecuadorian Pristimantis.
Article
Full-text available
We studied the historical prevalence of the invasive and pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) among amphibians from the Bolivian Andes. Our aim was also to determine its geographic pattern of dispersion, and a potential host taxonomic signature. We collected frog tissue samples from nine museum collections covering a period from 1863 to 2005 and from the field during 2009–2016. Bd was diagnosed via quantitative PCR in 599 individuals of 17 genera and 54 species. We found an overall Bd prevalence of 41% among 44 species tested. The first incidence of Bd was from a Telmatobius culeus in 1863; this is the earliest report of detection for this pathogen in the world. Results reveal a non-random historical and geographical pattern of Bd occurrence and amphibian declines that suggests the presence of two different invasive strains, an ancient endemic and a more recent introduction. Prevalence of Bd increased significantly by the mid-1990s, particularly in the cloud-forests, and this is coincident with the timing of drastic amphibian declines. In contrast, amphibians occurring in drier altiplano habitats have persisted in spite of Bd presence. We hypothesize that the early 1990s, and the cloud-forests in central Bolivia were the center of an epidemic surge of Bd that took its toll on many species, especially in the genus Telmatobius. Further sampling of cloud-forest species, and ongoing genetic studies of Bd isolates from Bolivia should help resolve the history of this invasive pathogen and test hypotheses on the differential response of endangered hosts.
Article
Full-text available
The spider diversity of the family Anyphaenidae in premontane, low evergreen montane and cloud forest from the Chocó region of Ecuador is examined. A total of 287 adult specimens were collected and 19 morphospecies were identifi ed based on male specimens. Thirteen new species are described and one new genus is proposed. Five new species are described in the genus Katissa Brescovit, 1997: Katissa kurusiki sp. nov., K. puyu sp. nov., K. tamya sp. nov., K. yaya sp. nov. and K. guyasamini sp. nov. The new genus Shuyushka gen. nov. is proposed and includes three species: Shuyushka achachay gen. et sp. nov., S. moscai gen. et sp. nov. and S. wachi gen. et sp. nov. Finally, fi ve species are described in the genus Patrera Simon, 1903: P. hatunkiru sp. nov., P. philipi sp. nov., P. suni sp. nov., P. shida sp. nov. and P. witsu sp. nov. New records are provided for Patrera fulvastra Simon, 1903 and Josa nigrifrons Simon, 1897.
Article
Full-text available
Significance Human influences are causing the disappearance of species at a rate unprecedented in millions of years. Amphibians are being particularly affected, and extinctions of many species may be inevitable. The Sierra Nevada yellow-legged frog ( Rana sierrae ) was once common in the mountains of California (United States), but human impacts have driven it near extinction. Repeated surveys of thousands of water bodies in Yosemite National Park show that the decline of R. sierrae has recently reversed and that population abundance is now increasing markedly in part because of reduced influence of stressors, including disease and introduced fish. These results suggest that some amphibians may be more resilient than is assumed, and with appropriate management, declines of such species may be reversible.
Article
Full-text available
Reportamos la presencia de nuevas poblaciones de sapos arlequines pertenecientes a las especies Atelopuspalmatus y A. nepiozomus en las provincias de Pastaza y Loja respectivamente. Estos nuevos hallazgos permiten re-evaluar y corroborar el estado de conservación de estas especies. La información aquí reportada es una contribución al conocimiento del género Atelopus sobre el cual existe todavía importantes vacíos de información sobre su taxonomía, sistemática e historia natural.
Article
Captive and wild frogs from North and Central America and Australia recently have died with epidermal infections by chytridiomycete fungi. We isolated a chytridiomycete into pure culture from a captive, blue poison dart frog that died at the National Zoological Park in Washington, D.C. Using this isolate, we photographed developmental stages on nutrient agar, examined zoospores with transmission electron microscopy, and inoculated test frogs. This inoperculate chytrid develops either monocentrically or colonially and has thread-like rhizoids that arise from single or multiple areas on the developing zoo-sporangium. The taxonomically important features of the kinetosomal region of the zoospore indicate that this chytrid is a member of the Chytridiales but differs from other chytrids studied with transmission electron microscopy. Its microtubule root, which begins at kinetosome triplets 9–1 and extends parallel to the kinetosome into the aggregation of ribosomes, is distinctive. Histologic examination of test frogs revealed that the pure culture infected the skin of test frogs, whereas the skin of control frogs remained free of infection. The fungus is described as Batrachochytrium, dendrobatidis gen. et sp. nov.