Kristofer M. Helgen; C. Miguel Pinto; Roland Kays; Lauren E. Helgen; Mirian T. N. Tsuchiya; Aleta Quinn; Don E. Wilson; Jesús E. Maldonado

“ We demonstrate that four olingo species can be recognized, including a Central American species (Bassaricyon gabbii) , lowland species with eastern, cis-Andean (Bassaricyon alleni) and western, trans-Andean (Bassaricyon medius) distributions, and a species endemic to cloud forests in the Andes. The oldest evolutionary divergence in the genus is between this last species, endemic to the Andes of Colombia and Ecuador, and all other species, which occur in lower elevation habitats. Surprisingly, this Andean endemic species, which we call the Olinguito, has never been previously described ”

“ The single exception was that the topology that recovered the node uniting Bassaricyon alleni and Bassaricyon medius to the exclusion of Bassaricyon gabbii was assigned a slightly lower Bayesian posterior probability value of 0.97, but all other methods lent full support to this topology (Bassaricyon gabbii, (Bassaricyon medius, Bassaricyon alleni) ) . These results were also well-supported by our comparisons of morphological characters and together lend strong support for this scenario as being an accurate representation of the evolutionary history of diversification within Bassaricyon. ”

“ Non-focal and outgroup taxa are shaded in gray, Bassaricyon species and subspecies are color coded, samples of Bassaricyon medius medius and Bassaricyon neblina neblina that were collected within 5 km of each other in Ecuador are shaded. The concordance of our recovered topology and estimates of genetic divergence with previous phylogenetic studies of the Procyonidae suggests that data from cytochrome b and CHRNA1 across sampled taxa have provided a well-supported framework in which the species relationships and divergence dates within Bassaricyon can be reliably assessed. ”

“ Given that Olinguitos are directly dependent on cloud forest for habitat and food, deforestation appears to be the primary threat to Olinguito populations, and this IUCN categorization reflects our concerns about habitat destruction across its relatively restricted geographic range. ”

“ Based on our distribution model, it appears that 42% of potential Olinguito habitat in Colombia and Ecuador has already been converted to agriculture or urban environments. Remaining habitat is highly fragmented and faces increasing threats from farming, grazing, deforestation for drug cultivation, logging, and climate change ( [137] , [260] , [262] , [263] , [264] ) . The long-term survival of Bassaricyon neblina will depend on the preservation of those upland forest fragments that remain, and restoration of degraded habitat to maintain connectivity between populations. ”

“ That the two earliest divergences within the genus involve what are today montane (Bassaricyon neblina of the Andes) or mostly montane (Bassaricyon gabbii of the Costa Rican, Nicaraguan, and western Panama highlands) taxa provides an indication that the isolation of upland Neotropical habitats was likely important in the early diversification of the genus. ”

“ Abstract AbstractWe present the first comprehensive taxonomic revision and review the biology of the olingos, the endemic Neotropical procyonid genus Bassaricyon, based on most specimens available in museums, and with data derived from anatomy, morphometrics, mitochondrial and nuclear DNA, field observations, and geographic range modeling. Species of Bassaricyon are primarily forest-living, arboreal, nocturnal, frugivorous, and solitary, and have one young at a time. ”

“ Colored arrows reflect dispersal events between ancestral and derived areas, with colors matching with recipient areas. Upper boxes: different dispersal constraints at time intervals 0–10 mya and 10–30 mya, the former to simulate the effect of the land bridge formation between Central and South America, the latter restricted dispersal due to the absence of the land bridge; the cells in green indicate no restriction to dispersal, cells in gray indicate a reduction by half in dispersal capability, and cells in black do not allow dispersal. ”

“ More recent field studies in Panama ( [53] , [236] , Figure 23) are limited but likewise show Bassaricyon medius to be nocturnal, arboreal, frugivorous (and nectarivorous) , and to feed in the same trees as kinkajous, which sometimes displace the smaller olingos. It is mostly solitary at night, spends its days in tree holes or other arboreal hide-outs, and usually has one young at a time. One olingo was found to have a home range of 37 hectares. Field workers have described this species’ vocalizations as “whey-chuck”, “wer-toll”, or “wake-up.” ”

“ This is the largest subspecies of Bassaricyon neblina, with a short rostrum, widely splayed zygomata, wide rostrum and braincase, and very large molars and posterior premolars; the dorsal pelage is of moderate length, tan to orangish-brown in overall color, with prominent black and gold tipping, with a more grayish face and limbs, with the limbs bearing relatively short fur, and a tail usually grizzled with golden-brown fur tipping. ”

“ The Loja-Huancabamba region of the Andes in southern Ecuador and northern Peru. Much remains to be learned about the Olinguito, including its distribution. The taxonomic description of this species is the first step toward further studies of its biology, and we look forward to future reports of additional discoveries from Andean cloud forests regarding this beautiful procyonid. ”

“ Maximum Parsimony, Maximum Likelihood and Bayesian analyses were performed for each gene and a concatenation of the two genes to check for any incongruence in structure and support of the Bassaricyon clade. All Bayesian and Maximum Likelihood phylogenetic inferences were carried out using the Cipres Portal ( [91] ) . Indels were treated as missing data or non-informative data for all of the analyses as in previous molecular phylogenetic studies of procyonids ( [71] ; [72] ) .
Pairwise distances for cytochrome b were generated using the Kimura 2-parameter model using MEGA4 ( [92] ) . ”

“ The fur of the face in front of the eyes is shorter and gray or buff with black tipping, sometimes with a pale cream ring around the eyes. The hairs of the tail are strongly tipped with gold, or with both golden and blackish-brown tipping. In contrast to specimens of other Bassaricyon, the tail is not conspicuously banded, though when viewed in the right light, a banding pattern of alternating golden and brown hues is weakly apparent in some specimens. A white terminal tail tip is present in a minority of individuals. ”

“ Because some olingo taxa demonstrate significant sexual dimorphism in cranial measurements, patterns of morphometric variation in males and females were compiled and analyzed separately. Principal Component Analysis (PCA) and Discriminant Function Analyses (DFA) were undertaken using a combination of cranial and dental measurements indicated in tables and in the text, selected to sample craniodental size and shape, and to maximize sample size. All measurement values were transformed to natural logarithms prior to multivariate analysis. ”

“ Distribution.
This species occurs in the central portion of Central America, in montane and foothill forests, from northern Nicaragua to Costa Rica and into the Chiriqui Mountains in western Panama, possibly also extending north into Honduras and Guatemala ( [29] ; see below) . Northern olingos are recorded at elevations as low as sea level, but are most commonly encountered in forests above 1000 m, and extend elevationally at least as high as 1700 m (USNM 324293) , and probably as high as the upper limit of forest on the highest peaks in Costa Rica. ”

“ Reagent volumes were the same as for cytochrome b amplification (above) , except 2μL of gDNA was added for CHRNA1 amplification, decreasing sterile water volume to 10.8μL. We were unable to sequence the nuclear intron from the turbinate bone sample.
Each PCR was conducted with negative and positive controls to minimize risk of spurious results from contamination or failure of the reaction. A 2μL sample of the PCR product was stained with ethidium bromide and run on an agarose gel with a 1 kb ladder. The gel was placed under UV light to visualize the PCR products. ”

“ The genetic-like algorithm was used to simultaneously find the topology, branch lengths and substitution model parameters with the greatest log-likelihood (lnL) for each dataset. Bootstrap support was generated with 1000 replicates and two independent searches per replicate.
jModeltest version 0.1.1 ( [95] ) was used to find the best model of sequence evolution. We chose to partition the cytochrome b data in order to minimize the number of parameters and to account for differences in base composition and substitution rates among the different codon positions ( [96] ) . ”

“ Colombia (tentatively attributed) : AMNH 37797, FMNH 69578.
Discussion Carnivore taxonomy
Descriptions of new species of carnivores are especially rare, and the order Carnivora is generally considered one of the most completely characterized groups across the entire tree of life ( [237] , [238] ) . Bassaricyon neblina is a deeply divergent lineage within its genus, a very morphologically distinctive member of the family Procyonidae, and even shows signs of evolutionary diversification across its geographic range. ”

“ Type specimens and localities.
The holotype of medius is BMNH 9.7.17.10, an adult male (skin and skull) from “Jimenez, mountains inland of Chocó, W. Colombia, 2400 feet” ( [227] ) .
The holotype of orinomus is USNM 179157, an adult male (skin and skull) , from “Cana (altitude 1,800 feet) , in the mountains of eastern Panama” ( [228] ) . [228] figured the holotype skull.
Diagnosis.
Bassaricyon medius is a medium-sized olingo, smaller (on average) than Bassaricyon gabbii of Mesoamerica and larger than Bassaricyon neblina of the Andes. ”

“ We report on the biology of this new species based on information from museum specimens, niche modeling, and fieldwork in western Ecuador, and describe four Olinguito subspecies based on morphological distinctions across different regions of the Northern Andes.
Introduction
“New Carnivores of any sort are always few and far between...”
Oldfield Thomas (1894:524)
Olingos (genus Bassaricyon J.A. Allen, 1876) are small to medium-sized (0.7 to 2 kg) arboreal procyonids found in the forests of Central America and northern South America. ”

“ FMNH 70723, adult male, skin, skull, and postcranial skeleton, Guapantal, 2200 m, Urrao, Antioquia Department, Colombia (collected by P. Hershkovitz, 28 April 1951) .
Reproductive isolation and genetic divergence ofBassaricyon neblina
Information from sympatric occurrences and captive breeding demonstrates that the Olinguito, Bassaricyon neblina, is reproductively isolated from other species of Bassaricyon and clearly constitutes a distinct “biological species” (i.e., sensu [140] , [141] ) . ”

“ Gazetteers published by [110] , [111] , [112] ) , [113] , [114] ) , [115] , [116] , [117] , and [118] were especially helpful in georeferencing Neotropical expedition and collecting localities represented in museum collections.
We used Maximum Entropy Modeling (Maxent) ( [119] ) to predict the geographic range of the geographic range of the four Bassaricyon species at broad scales based on vouchered localities (Appendix 2) and 20 environmental variables representing potential vegetation and climate. ”

“ It differs from other Bassaricyon in its smaller body and cranial size; longer, denser, and more richly coloured dorsal pelage (black-tipped, tan to strikingly orange- to reddish-brown) ; indistinctly banded, bushier, and proportionally shorter tail (at least compared to the lowland olingos, Bassaricyon alleni and Bassaricyon medius, Table 5) ; (externally) more rounded face with a blunter, less tapering muzzle; smaller and more heavily furred external ears, and considerably reduced auditory bullae, with a markedly smaller external auditory meatus ”

“ Scale bar = 50 mm.Skulls of adult female Bassaricyon. From left to right: Bassaricyon gabbii (AMNH 140334, Lajas Villa, Costa Rica) ; Bassaricyon medius orinomus (AMNH 37797, Puerta Valdivia, Antioquia District, Colombia) ; Bassaricyon alleni (FMNH 86908, Santa Rita, Rio Nanay, Maynas, Loreto Region, Peru) ; Bassaricyon neblina hershkovitzi (FMNH 70727, San Antonio, Agustin, Huila District, Colombia) . Scale bar = 50 mm. Table 3. "Cranial measurements for olingo species (compiled separately by sex) . For each measurement, means are provided, ± standard deviation, with ranges in parentheses." ”

“ Forested areas above 1000 m in Central America can be understood to be the core distribution of this species. Vouchered records are from the north-central mountains of Nicaragua ( [166] , AMNH, USNM) ; the mountains of Costa Rica, including the slopes stretching down to the Atlantic coast ( [167] , [166] , [169] , [171] , [172] , [173] , [28] , [174] , [175] , [176] , [29] ) and a few records of observations from the Pacific slopes (Puntarenas Province: [177] ; Guanacaste Province: [178] ) ; and in the Chiriqui Mountains of western Panama ( [170] , ANSP, USNM) . ”

“ Molecular data for Bassaricyon neblina from a cell line were first generated and used in a phylogenetic study of carnivore relationships by [123] , [124] ) , apparently the same animal whose karyotype was reported and discussed by [147] (also [148] ) . DNA sequence data (12S rRNA, cytochrome b) from this sample, available on Genbank, have been used in various other published studies (e.g., [156] , [157] , [158] , [159] , [160] , [161] , [162] , [87] , [163] , [164] , [122] , but not in some important studies, e.g., [71] , [165] ) . ”