Research Article |
Corresponding author: Adolfo Amézquita ( aamezquita@bioconservancy.org ) Academic editor: Anthony Herrel
© 2024 Adolfo Amézquita, Fernando Vargas-Salinas, Iván Ramos, Pablo Palacios-Rodríguez, Erika Nathalia Salazar, Michelle Quiroz, Wilmar Bolívar, Diana M. Galindo-Uribe, Luis A. Mazariegos-H.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Amézquita A, Vargas-Salinas F, Ramos I, Palacios-Rodríguez P, Salazar EN, Quiroz M, Bolívar W, Galindo-Uribe DM, Mazariegos-H LA (2024) Molecular phylogenetics uncovers two new species in the genus Phyllobates (Anura, Dendrobatidae): the terrible frog gets two new sisters. ZooKeys 1212: 217-240. https://doi.org/10.3897/zookeys.1212.126733
|
True poison-dart frogs (Phyllobates, Dendrobatidae) evolved the ability to secrete batrachotoxins, the most powerful alkaloids known to date. The genus comprises five species whose systematics, at first glance, appeared clear. The most derived clade would include two Colombian species (P. terribilis and P. bicolor) with the highest toxicity, the largest body size, and predominantly yellow body colouration. The other three species (P. aurotaenia, P. vittatus, and P. lugubris) are less toxic on average, have smaller size, and are predominantly black with bright dorsolateral stripes. Recent research has revealed the existence of two major lineages among the three Colombian species. The northern lineage appears to result from a complex evolutionary history, including perhaps introgression among yellow and black taxa. The southern lineage instead revealed the existence of new clades closely related to P. terribilis, black and yellow, that arguably deserve their recognition as new species. Here, available evidence is combined to support the erection of southern populations of P. aurotaenia as a new highly toxic species, sister to P. terribilis, and much closer to it than to any other yellow or black-bodied species, Phyllobates samperi sp. nov. Their common ancestor is sister to an additional yellow species, which we also describe here as Phyllobates bezosi sp. nov. Both new species can be externally diagnosed using colouration. Our previous and current analyses also suggest the existence of additional taxa and corroborate multiple transitions in colouration across these hypertoxic taxa.
Las verdaderas ranas de dardo venenoso (género Phyllobates) evolucionaron la capacidad de secretar batracotoxinas, los alcaloides más poderosos que se conocen hasta hoy. El género comprende cinco especies, cuya taxonomía parecía clara a primera vista. El clado más derivado incluye dos especies colombianas (P. terribilis and P. bicolor) con la mayor toxicidad, mayor tamaño y coloración predominantemente amarilla. Las otras tres especies (P. aurotaenia, P. vittatus and P. lugubris) son en general menos tóxicas, menores en tamaño y predominantemente negras con conspicuas manchas dorsolaterales.Una reciente investigación confirmó la existencia dos linajes mayores entre las tres especies colombianas. El del norte parece ser el producto de una historia evolutiva compleja, incluyendo quizás introgresión entre especies amarillas y negras. El del sur reveló en cambio la existencia de nuevos clados filogenéticamente cercanos a P. terribilis, uno negro y dos amarillos, que requieren reconocimiento formal como nuevas especies. Esta investigación combina la evidencia disponible para apoyar el reconocimiento de las poblaciones del sur de P. aurotaenia como una nueva especie negra, hermana de P. terribilis, y mucho más cercana a ella que a las otras especies negras o amarillas, Phyllobates samperi sp. nov. Su ancestro común es a su vez el grupo hermano de una especie adicional, amarilla, que también describimos como Phyllobates bezosi sp. nov. Las dos nuevas especies pueden ser externamente reconocidas por la coloración. Nuestros análisis previos y actuales también sugieren la existencia de otras especies no descritas y corroboran la existencia de múltiples transiciones de coloración en la evolución de las especies hipertóxicas.
Chocoan region, Colombia, hypertoxic frogs, molecular phylogenetics, Phyllobates
Chocó biogeográfico, Colombia, filogenética molecular, Phyllobates, ranas hipertóxicas
True poison dart frogs (Phyllobates, Dendrobatidae) were discovered by Chocó indigenous people in western Colombia and used to poison blowgun darts by rubbing them on the frogs’ backs (
The genus Phyllobates sensu
The evolutionary relationships between the three Colombian species appeared clear at first glance (
It was suggested very early that P. aurotaenia may consist of two clades: the narrow-banded and broad-banded forms, differing in colouration and, perhaps, altitudinal distribution (
In the course of field investigations on the evolution of hypertoxicity in poison frogs, we noticed unexpected genetic and colouration traits in a population currently assigned to P. aurotaenia. Moreover, we acknowledged the existence of another distinctive form, predominantly yellow in colouration, which has not been genotyped before. Both observations led us to question the taxonomic status of the involved lineages. A more recent analysis of gene flow among the Colombian species and populations of Phyllobates (
To the north, conspicuous geographic variation in colouration (
The specimens used in this study were collected during fieldwork conducted between 2009 and 2022 in localities of the Colombian Chocó bioregion, where Phyllobates species occur. Specimens of the black clade were collected around Buenaventura, Magüipi, Chucheros, and San Cipriano, all within the Departamento del Valle (Colombia). Specimens of the yellow clade were collected around the Garrapatas river, near the border between Departamento del Chocó and Departamento del Valle del Cauca. We list further details in species descriptions.
After capture, most specimens were photographed and measured (snout-to-vent length, SVL); some were then released at the site of capture. A number were transported to the laboratory, euthanised using topical lidocaine, fixed in 10% formalin, and stored in 70% ethanol, to be deposited at the Amphibian Collection of the C.J. Marinkelle Museum of Natural History at Universidad de los Andes (
To reproduce the phylogenetic hypothesis on the studied taxa (
Our matrix thus included three mitochondrial loci (16S rRNA, COI, and Cytb) using primers 16Sar and 16Sbr (
Homologous sequences of each region were aligned using MAFFT under default parameters (
To estimate the degree of genetic divergence among the focal Colombian lineages, we used a subset of 42 (16S rRNA), or 43 (COI and Cytb) sequences. We chose these individuals because we had a complete set of DNA sequences for all three genes, with one exception (Suppl. material
Using a digital calliper, we took the following measurements of the preserved adults: length from snout to vent (SVL), femur length from flexed knee to groin (FL), tibia length between heel and outer surface of flexed knee (TL), greatest body width, measured under armpits (GBW), head width between angles of jaws (HW), head length from tip of snout to angle of jaw (sagittal; HL), horizontal eye length (EL), internarinal distance (IN), interorbital distance (IOD); distance from centre of naris to anterior edge of the eye (NED), horizontal diameter of the tympanum (HDT), left-hand length from palmar tubercle to longest (third) finger (HaL), and left foot length from inner metatarsal tubercle to longest (fourth) finger (FoL). For tadpoles, we measured the body length (BL) and tail length (TaL) following
Two of the captured males of the black clade were found carrying four back-riding tadpoles; the eight tadpoles were euthanised in a lidocaine solution upon capture, preserved in 95% ethanol, and stored in 70% ethanol. Moreover, a pair of frogs produced a fertile egg clutch while housed in the laboratory; one captive-born tadpole was raised until Gosner’s developmental stage 31 (
We recorded at least seven spontaneous advertisement calls for each of ten males of the black clade at three localities: Piangüita (n = 8 males, including the holotype), Bahía Málaga (n = 1) and Bajo Calima (n = 1), all in the vicinities of Buenaventura city, Colombia. Calls were recorded using a Marantz PMD660 digital recorder, at 16-bit resolution and 44.1 kHz sampling rate, and a unidirectional microphone (AKG D-190-E, Shure BG4.1, or Sennheiser ME-62/K6), positioned at a distance of 0.5–2.0 m in front of the male. We also recorded two and seven calls of two males of the yellow clade using the same equipment and parameters under laboratory conditions. Body temperature was measured after recording with an infrared thermometer positioned at ~ 10 cm of male dorsum.
To allow for proper across-species comparisons in the future, we recognise a homologous sound unit as the sound produced by a single thoracic compression (
The final dataset consisted of 1926 mtDNA nucleotides (16S: 568 bp; COI: 658 bp; Cytb: 700 bp) aligned for 103 individuals and including 425 parsimony-informative and 68 singleton sites. The mtDNA analysis essentially reproduced the previously published phylogenetic hypothesis either based on RNA extracts of the whole genome or in mtDNA alone (
Recovered relationship between Colombian species and clades of Phyllobates and proposed taxonomic arrangement in the light of current evidence. Green dots denote nodal support (UFBoot2) of at least 95%. The individuals outlined by two-coloured bars remain without a name, because they represent the probable result of introgression or hybridisation (
The pairwise genetic distances between each of the new species and all other species further corroborated their status as independent lineages (Table
Two-dimensional representation of genetic (16S, COI, and Cytb) uncorrected P-distances between clades of Colombian Phyllobates, including the two new species described herein. Two distinctive forms of P. aurotaenia are shown: the Baudo West and Atrato River forms in
Two-dimensional distribution of genetic (16S, COI, and Cytb) distances between individuals of the same or different operational taxa addressed in this research. To read each panel, start from the panel title and then use the colour legend to identify the kind of comparison depicted by each uniquely coloured set of data points. To facilitate reading, only comparisons from species in the southern lineage are shown, which includes the newly described species.
Genetic distances. Uncorrected p-genetic (16S, COI, and Cytb) distances among species and operational taxa of Phyllobates addressed in this study. Standard errors (95%) were estimated from 999 bootstraps and are presented above diagonal.
Clade | auAt | auBw | bezo | bico | sp | samp | terr |
---|---|---|---|---|---|---|---|
16S | |||||||
P. aurotaenia At | 0.005 | 0.006 | 0.004 | 0.005 | 0.007 | 0.007 | |
P. aurotaenia Bw | 0.011 | 0.007 | 0.004 | 0.007 | 0.008 | 0.007 | |
P. bezosi sp. nov. | 0.018 | 0.025 | 0.007 | 0.005 | 0.006 | 0.005 | |
P. bicolor | 0.009 | 0.010 | 0.023 | 0.006 | 0.008 | 0.007 | |
P. sp. (bicolor South) | 0.013 | 0.020 | 0.012 | 0.018 | 0.006 | 0.006 | |
P. samperi sp. nov. | 0.025 | 0.027 | 0.017 | 0.028 | 0.018 | 0.004 | |
P. terribilis | 0.022 | 0.026 | 0.014 | 0.026 | 0.015 | 0.009 | |
COI | |||||||
P. aurotaenia At | 0.006 | 0.009 | 0.006 | 0.010 | 0.006 | 0.010 | |
P. aurotaenia Bw | 0.020 | 0.010 | 0.007 | 0.010 | 0.007 | 0.010 | |
P. bezosi sp. nov. | 0.047 | 0.055 | 0.007 | 0.007 | 0.008 | 0.007 | |
P. bicolor | 0.039 | 0.044 | 0.042 | 0.005 | 0.006 | 0.007 | |
P. sp. (bicolor South) | 0.057 | 0.060 | 0.031 | 0.033 | 0.009 | 0.006 | |
P. samperi sp. nov. | 0.032 | 0.037 | 0.044 | 0.042 | 0.058 | 0.009 | |
P. terribilis | 0.063 | 0.064 | 0.029 | 0.045 | 0.024 | 0.058 | |
Cytb | |||||||
P. aurotaenia At | 0.005 | 0.011 | 0.006 | 0.009 | 0.010 | 0.010 | |
P. aurotaenia Bw | 0.016 | 0.011 | 0.005 | 0.009 | 0.010 | 0.010 | |
P. bezosi sp. nov. | 0.085 | 0.076 | 0.010 | 0.009 | 0.008 | 0.007 | |
P. bicolor | 0.026 | 0.016 | 0.074 | 0.008 | 0.010 | 0.010 | |
P. sp. (bicolor South) | 0.058 | 0.051 | 0.054 | 0.048 | 0.008 | 0.008 | |
P. samperi sp. nov. | 0.078 | 0.071 | 0.041 | 0.071 | 0.049 | 0.005 | |
P. terribilis | 0.076 | 0.069 | 0.035 | 0.068 | 0.045 | 0.018 |
P. aurotaenia:
P. aurotaenia:
Holotype
• Adult male. Field original label: 16B. Museum ID:
Morphometry. Body measurements and sex of collected individuals of the new species that are described herein. HT denotes holotype. See Materials and methods for abbreviations.
Species | ID | Sex | SVL | FL | TL | GBW | HW | HL | EL | IN | IOD | NED | HDT | HaL | FoL |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
P. samperi sp. nov. |
|
M | 24.0 | 11.8 | 13.7 | 9.0 | 8.3 | 9.3 | 3.3 | 3.2 | 5.1 | 2.4 | 2.8 | 6.2 | 10.4 |
P. samperi sp. nov. | UVC-H 7878 | F | 27.0 | 11.2 | 13.6 | 8.2 | 8.6 | 9.2 | 3.4 | 3.6 | 5.8 | 3.2 | 1.8 | 6.5 | 12.5 |
P. samperi sp. nov. | UVC-H 7879 | M | 21.7 | 9.0 | 11.5 | 7.7 | 6.5 | 7.6 | 3.5 | 2.4 | 5.9 | 2.4 | 1.4 | 5.7 | 9.7 |
P. samperi sp. nov. | UVC-H 8849 | – | 23.1 | 9.5 | 11.5 | 7.2 | 7.4 | 8.1 | 3.3 | 3.7 | 5.0 | 2.4 | 1.1 | 6.1 | 10.9 |
P. samperi sp. nov. | UVC-H 15332 | M | 21.5 | 8.0 | 11.6 | 6.9 | 6.4 | 8.5 | 2.7 | 3.2 | 4.2 | 2.5 | 1.9 | 5.5 | 9.4 |
P. samperi sp. nov. |
|
M | 26.5 | 10.6 | 12.7 | 7.4 | 8.9 | 8.0 | 3.1 | 2.7 | 5.8 | 2.7 | 1.3 | 5.8 | 9.5 |
P. samperi sp. nov. |
|
– | 25.9 | 9.3 | 12.8 | 8.1 | 7.8 | 8.0 | 2.7 | 2.7 | 4.4 | 2.5 | 1.2 | 5.6 | 11.1 |
P. samperi sp. nov. |
|
– | 26.1 | 10.8 | 13.9 | 8.9 | 7.7 | 9.2 | 3.1 | 2.8 | 5.7 | 2.6 | 1.1 | 6.1 | 11.0 |
P. samperi sp. nov. |
|
M | 25.6 | 10.6 | 12.6 | 9.2 | 7.7 | 8.0 | 2.5 | 3.0 | 4.7 | 2.5 | 1.2 | 6.2 | 11.0 |
P. samperi sp. nov. |
|
M | 25.4 | 9.2 | 11.2 | 9.3 | 8.9 | 9.3 | 3.9 | 2.8 | 4.6 | 2.6 | 1.2 | 5.8 | 10.7 |
P. samperi sp. nov. |
|
M | 25.7 | 9.2 | 12.8 | 9.3 | 8.0 | 8.0 | 3.9 | 3.0 | 5.7 | 2.5 | 1.1 | 5.7 | 11.1 |
P. samperi sp. nov. |
|
M | 24.1 | 12.3 | 12.4 | 7.7 | 7.5 | 9.2 | 3.6 | 2.4 | 5.7 | 2.0 | 2.8 | 7.1 | 12.4 |
P. samperi sp. nov. |
|
M | 25.6 | 10.2 | 13.8 | 9.8 | 8.6 | 9.9 | 3.8 | 3.7 | 5.9 | 2.5 | 2.8 | 6.4 | 11.2 |
P. samperi sp. nov. |
|
M | 25.9 | 11.6 | 12.4 | 8.7 | 7.6 | 9.5 | 3.5 | 3.4 | 6.4 | 2.4 | 2.8 | 6.3 | 10.0 |
P. samperi sp. nov. |
|
M | 25.0 | 11.2 | 14.4 | 7.2 | 7.6 | 8.5 | 3.3 | 3.7 | 5.8 | 3.6 | 2.7 | 6.5 | 11.0 |
P. samperi sp. nov. |
|
F | 27.0 | 10.7 | 13.7 | 8.6 | 8.2 | 9.0 | 3.2 | 3.0 | 5.5 | 2.5 | 2.9 | 6.4 | 11.9 |
P. samperi sp. nov. |
|
M | 24.5 | 11.3 | 13.6 | 7.1 | 7.2 | 8.7 | 3.3 | 4.3 | 5.4 | 2.4 | 2.8 | 6.1 | 11.8 |
P. samperi sp. nov. |
|
– | 24.5 | 11.7 | 12.3 | 7.1 | 7.4 | 9.9 | 3.3 | 3.0 | 5.2 | 2.6 | 2.8 | 5.8 | 11.9 |
Mean | 25.0 | 10.5 | 12.8 | 8.2 | 7.8 | 8.8 | 3.3 | 3.1 | 5.4 | 2.6 | 2.0 | 6.1 | 11.0 | ||
SD | 1.6 | 1.2 | 1.0 | 0.9 | 0.7 | 0.7 | 0.4 | 0.5 | 0.6 | 0.3 | 0.8 | 0.4 | 0.9 | ||
P. bezosi sp. nov. | AA_7242 | – | 32.9 | 15.5 | 17.0 | 13.0 | 11.5 | 8.9 | 4.8 | 3.3 | 4.1 | 3.2 | 3.1 | 10.4 | 16.2 |
P. bezosi sp. nov. | AA_7243 | – | 32.9 | 16.3 | 17.0 | 13.4 | 11.2 | 12.1 | 4.8 | 3.6 | 4.3 | 3.6 | 3.4 | 10.2 | 16.7 |
P. bezosi sp. nov. | AA_7244 | – | 35.3 | 15.4 | 16.7 | 12.2 | 11.6 | 10.5 | 4.5 | 3.8 | 4.9 | 4.7 | 3.2 | 9.4 | 17.4 |
P. bezosi sp. nov. | AA_7245 | – | 30.9 | 14.0 | 16.6 | 12.0 | 12.9 | 7.5 | 4.6 | 4.3 | 7.5 | 4.1 | 2.8 | 9.1 | 14.2 |
P. bezosi sp. nov. | CPZ-UV 9191 (HT) | M | 32.6 | 16.2 | 17.6 | 10.9 | 10.8 | 9.4 | 4.8 | 4.5 | 3.4 | 3.1 | 2.6 | 8.9 | 15.2 |
P. bezosi sp. nov. | CPZ-UV 9192 | M | 34.2 | 16.4 | 18.5 | 11.1 | 11.6 | 10.5 | 5.2 | 5.3 | 3.3 | 3.6 | 2.6 | 10.2 | 16.4 |
P. bezosi sp. nov. | CPZ-UV 9193 | M | 35.0 | 17.1 | 18.2 | 12.2 | 12.3 | 10.5 | 5.0 | 5.2 | 3.4 | 3.1 | 2.6 | 9.2 | 16.9 |
P. bezosi sp. nov. | CPZ-UV 9194 | M | 32.3 | 16.0 | 17.6 | 12.0 | 10.9 | 9.3 | 5.2 | 5.2 | 3.5 | 3.1 | 2.7 | 8.9 | 14.7 |
P. bezosi sp. nov. | CPZ-UV 9195 | F | 36.1 | 17.6 | 19.7 | 13.1 | 12.1 | 10.8 | 5.4 | 5.7 | 4.3 | 3.6 | 3.0 | 10.1 | 17.0 |
Mean | 33.6 | 16.0 | 17.6 | 12.2 | 11.7 | 9.9 | 4.9 | 4.5 | 4.3 | 3.6 | 2.9 | 9.6 | 16.1 | ||
SD | 1.7 | 1.0 | 1.0 | 0.9 | 0.7 | 1.3 | 0.3 | 0.9 | 1.3 | 0.5 | 0.3 | 0.6 | 1.1 |
Phyllobates samperi sp. nov. is a small to medium-sized dendrobatid with an adult SVL of 21–27 mm (24.5 ± 1.6 mm, mean ± SD, n = 17). It is mostly pitch-black, with a narrow golden yellow/orange complete dorsolateral stripe (sensu
Phyllobates samperi sp. nov. can be distinguished from other members of its genus as follows: colouration and size easily separate P. samperi sp. nov. from P. terribilis, P. bicolor, and P. bezosi sp. nov. (Fig.
The holotype is an adult male, 24.0 mm in snout-to-vent length, not dissected but assumed to be sexually mature because it was recorded (Fig.
Advertisement calls. Sonagram of one series of advertisement calls (a trill, above) of the holotype male specimens of Phyllobates samperi sp. nov. and P. bezosi sp. nov. An oscillogram (dark lines below the upper sonogram) shows a typical pattern of increase in sound pressure level. A power spectrum (red lines to the right of the upper sonograms) reveals most call energy is concentrated. In the sonograms below, a detail is presented of three uni-note calls per species.
The background colour of the head, back, sides, vent, and limbs is pitch black. A narrow golden yellow/orange dorsolateral stripe extends from the tip of the snout, along the outer margin of the upper eyelid, to the dorsum near the dorsal base of the thigh. In some individuals, a few tiny orange or greenish freckles are present on the dorsal surface of limbs and arms and near the dorsal surface of cloacae (Fig.
The following description is based on a single individual raised until stage 31 (
Larvae. Developmental
ID | Stage | BL | TaL | ToL |
---|---|---|---|---|
|
31 | 8.04 | 13.98 | 22.02 |
|
25 | 3.27 | 5.97 | 9.24 |
|
25 | 3.41 | 6.1 | 9.59 |
|
25 | 2.65 | 6.19 | 8.84 |
|
25 | 3.16 | 6.87 | 10.03 |
|
25 | 3.47 | 6.82 | 10.29 |
|
25 | 4.1 | 6.69 | 10.79 |
|
25 | 3.8 | 6.74 | 10.54 |
|
25 | 3.68 | 6.68 | 10.36 |
Mean* | 3.44 | 6.52 | 9.96 | |
SD* | 0.44 | 0.35 | 0.68 |
Phyllobates samperi sp. nov. utters trill calls. Each trill (Fig.
Phyllobates samperi sp. nov. is a diurnal species that inhabits the understory of tropical humid forests in southwestern Colombia. From our observations, it thrives well in forests with moderate degrees of disturbance. Adults are found mainly on the leaf litter, near fallen logs, roots, large leaves, and other objects that provide refuge and probably food. Males defend territories by uttering advertisement calls usually from the leaf litter and less commonly from low perches; they sit near palm leaves, logs and roots, and often call while concealed under them. We observed very bold and aggressive responses when playing call recordings within males’ territories, including one individual hopping onto a researcher’s leg while looking for the invader. Males carry tadpoles on their backs and deposit them in ground-level phytotelmata, such as fallen palm bracts or leaf sheaths, or in semi-permanent puddles formed by rainwater on the forest floor. All of the collected back-riding tadpoles were at stage 25, matching the observation by
Colour variation among individuals of Phyllobates bezosi sp. nov. (HT: holotype) including A dorsal and B ventral images for adult individuals and C three froglets, which exhibit the typical colouration with dorsolateral lines that characterises juvenile Phyllobates. To compare with juveniles of this species, we add a single image D of P. samperi sp. nov.
The colour pattern of Pristimantis gaigeae (Anura: Craugastoridae) closely resembles that of Phyllobates samperi sp. nov. when found in sympatry. The same resemblance has also been reported for populations of Pristimantis gaigeae sympatric with Phyllobates lugubris and Phyllobates aurotaenia (Myers and Daly, 1983). Since Pristimantis gaigeae is not known to be toxic, the resemblance may have resulted from Batesian mimicry, but the topic requires to be studied.
In the course of a parallel project on Phyllobates, one of the co-authors (P. Palacios-Rodríguez) visited the type locality of P. aurotaenia (Peñalisa, in the upper Condoto River) three times between 2014 and 2015. The site was severely degraded due to coca crops and gold mining and, despite intensive search of four people in three different years, no individuals of Phyllobates were seen or heard. We studied P. aurotaenia individuals from El Dos locality, 15 km north of the type locality, which exhibit the narrow greenish band that characterises P. aurotaenia sensu lato (this study;
To the best of our knowledge, the new species is distributed in the wet forests along the lower San Juan and Dagua river basins in the vicinities of Buenaventura, Valle del Cauca, Colombia, extending eastwards to the foothills of the Cordillera Occidental, but not surpassing elevations above 200 m a.s.l. (Fig.
Systematics and estimated distribution of the seven Phyllobates species. Distribution was estimated from GBIF records (GBIF2023a–f), some of which were excluded based on well-acknowledged information (e.g., the genus distribution in South America is cis-Andean and not trans-Andean) as well as studies addressing distribution with the support of molecular data (
Despite the uncertain northern limit of P. samperi sp. nov., the potential distribution range does not surpass 20,000 Km2. It is currently known from seven localities, all enclosed within a ~ 1000 km2 polygon, where deforestation due to agriculture, the expansion of urban Buenaventura, and gold mining operations will certainly reduce the amount and quality of habitat shortly. Based on these criteria, we propose listing P. samperi sp. nov. as Vulnerable (VU: B1a, biii, iv) under the IUCN Red List of Threatened Species, but this status should be re-evaluated as further information, especially on the species’s range, becomes available.
Uramba Bahía Málaga National Park is a marine reserve recently created by the Colombian Government to protect the marine, coastal and estuarine habitats of Bahía Málaga, just north of Buenaventura. Five of the localities presented in this study, including the type locality, are close to the protected coast of Bahía Málaga and should be, at least to some extent, protected by the presence of the reserve. Other fractions of the species’ range are within smaller forest reserves, such as the San Cipriano Forestal Protective Reserve, which allows for moderate extractive activities (e.g., logging). Most of this species’s range is, nevertheless, outside protected areas, and therefore still exposed to threats.
The species is named honouring Colombian biologist Cristian Samper, for his lasting impact on the world of conservation science and environmental stewardship. As founding director of the Alexander von Humboldt Biodiversity Institute of Colombia, President and CEO of the Wildlife Conservation Society, Director of the Smithsonian National Museum of Natural History and Managing Director of the Bezos Earth Fund, Samper is a leading voice in global conservation efforts, overseeing initiatives that span across 65 countries to protect 11.6 million square kilometres of wild places. Samper’s expertise in biodiversity and policy has been instrumental in shaping international dialogues around conservation, making him a respected figure in the intersection of science, governance, and activism.
Holotype
• Adult male. Field original label: AA_7256. Museum ID: CPZ-UV 9191. Type locality in Colombia, Valle del Cauca, municipality of Bolívar, valley of the Garrapatas river, 4.43°, -76.43°, 700 m elevation, riverine rainforest, amidst the leaf litter, July 2023. Collected by Wilmar Bolívar and Adolfo Amézquita. Advertisement calls of this male (Fig.
Phyllobates bezosi sp. nov. is a medium-sized dendrobatid with an adult snout-to-vent length (SVL) of 31–36 mm (33.6 ± 1.7 mm, mean ± SD, n = 9). Body dorsal colouration is predominantly orange to yellow, with mid-dorsal black bands or blotches, usually incomplete and poorly defined, more frequently near the urostyle or between the eyes (Fig.
Phyllobates bezosi sp. nov. can be externally distinguished from other yellow or orange species of its genus by the prevalence of black in the ventral colouration (Fig.
The holotype is an adult male, 32.6 mm in snout-to-vent length, not dissected but assumed to be sexually mature because it was observed calling and recorded in captivity (Fig.
Adult individuals of P. bezosi sp. nov. are predominantly orange to yellow in the dorsum, more often with mid-dorsal black bands or blotches that are incomplete. The orange/yellow is more often replaced by predominantly black on the flanks, limbs, and ventral surfaces. A few individuals do not bear the middorsal black bands (Fig.
Phyllobates bezosi sp. nov. males utter trill calls (Fig.
During its time in the laboratory, a couple bred. A male was found carrying nine tadpoles on his back (7 June 2023). After placing him in another container with water, tadpoles were found free within 24 h (June 8). They were kept in a 25 × 20 × 20 cm aquarium, with algae, aquatic vegetation and supplemented with fish food. The first metamorph was found after 74 days (21 August) and the last one after 94 days (September 10). They expressed very early the two dorsolateral yellow bands that characterise the colouration of Phyllobates juveniles (Fig.
The species is known so far from a single locality: the type locality at the Garrapatas River (municipality of Bolívar, Valle del Cauca). The information on the status of the forests is very poor. Nonetheless, this area has suffered strong and increasing violence during the last decade, mostly associated with intensive illegal gold mining and narcotraffic. At the moment of writing this manuscript, access to the area is restricted by illegally armed groups that also constrain the movement of local inhabitants. Besides the aforementioned illegal activities, the lack of governmental authorities in the area and the long history of dendrobatid poaching in Valle del Cauca and Chocó allow us to infer a high risk for the species described herein. We thus propose to declare it Endangered EN B1ab(iii), B2ac(iii), under the IUCN Red List criteria (
The species is named honouring American entrepreneur Jeff Bezos, for his contributions to environmental conservation, particularly through the Bezos Earth Fund. The fund aims to combat climate change and preserve the natural world by funding scientists, NGOs, and other local communities dedicated to environmental action. The fund’s investments in conservation, restoration initiatives, food systems, clean energy, and sustainable development reflect a commitment to stewarding the planet for future generations. The considerable financial resources allocated for environmental causes symbolise an important step in mobilising private capital for public good, particularly in an era urgently demanding climate solutions. The name of this new remarkable species is expected to highlight this hallmark decision.
The Colombian Pacific lowlands are under strong pressure by the growth of illegal activities. Illegal mining destroys the riverine forests and riverbeds at unprecedented rates, and contaminates the water. Besides, the co-occurrence of gold mining with illegal crops favours the settlement and control of activities and routes by illegally armed groups. One of these activities is the poaching of dendrobatid frogs. Under this scenario, scientific and political decisions are badly needed to improve the survival chances of organisms therein. The study and acknowledgement of evolutionary unique lineages should decisively contribute to the quality of decisions in conservation.
To improve the coherence between previous molecular studies (
The three species of the southern Colombian clade now include two lowland and latitudinally separated species (Fig.
The taxonomic uncertainty is higher for the remaining populations of P. aurotaenia that together with P. bicolor form the northern clade of Colombian Phyllobates. Recent molecular (
Finally, the phylogenetic hypothesis and taxonomic arrangement presented here corroborate an interesting pattern observed in previous studies (
The authors would like to thank L.M. Arenas, D. Mejía, S.V. Flechas, J.A. Hernández, B.E. Velázquez, C. Esquivel, M.C. González for their assistance during fieldwork. We also thank T. Escovar and I. Medina for initial DNA extractions and for calling our attention to the existence of a separate clade; I. Medina, R. Márquez and J.M. Daza for support with phylogenetic analyses; and L.A. Barragán for assistance in archiving museum specimens. Frog drawings were mostly prepared by L.M. Arenas. Collections for this study were authorised by Biological Diversity Scientific Research Permits No. 2194 and 1070, granted respectively to the Universidad de los Andes and the Universidad del Valle by the Colombian Authority of Environmental Licenses (ANLA).
The authors have declared that no competing interests exist.
No ethical statement was reported.
Funding for this work was provided by a Seed Grant from the Faculty of Sciences at Universidad de los Andes (Bogotá) to AA and a Basic Sciences Research Grant from the Vice-rectory of Research at the same institution.
Conceptualization: AA, LAMH. Data curation: ENS, MQ, DMGU, AA, FVS, PPR, WB. Formal analysis: ENS, AA. Investigation: IR, WB, PPR, DMGU. Resources: LAMH, WB, MQ, IR. Visualization: FVS, LAMH. Writing - original draft: AA. Writing - review and editing: IR, LAMH, FVS, WB, PPR, ENS, MQ, DMGU.
Adolfo Amézquita https://orcid.org/0000-0001-7320-1143
Michelle Quiroz https://orcid.org/0000-0002-6955-4605
Luis A. Mazariegos-H https://orcid.org/0000-0003-4882-0414
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Linked data for primary biodiversity data
Data type: xlsx