The authors have declared that no competing interests exist.
Conceived and designed the experiments: JAH TBH KMD CCG ASB JLF AV EJS AJT. Contributed reagents/materials/analysis tools: JAH TBH KMD CCG ASB JLF ME AV EJS AJT. Wrote the paper: JAH KMD CCG ASB JLF TBH EJS AJT. Initial discovery and data collection: JAH TBH ME AV. Collected and analyzed the molecular data: JAH ME AV KMD ASB AJT. Collected and analyzed the morphological data: JAH ME AV CCG EJS. Collected and analyzed the behavioral and ecological data: JAH TBH AV JLF. Establishment of field site and proposed national park: JAH TBH ME.
In June 2007, a previously undescribed monkey known locally as “lesula” was found in the forests of the middle Lomami Basin in central Democratic Republic of Congo (DRC). We describe this new species as
Discoveries of new African primate species are rare but significant events that clarify taxonomic and evolutionary relationships and highlight important regions of biodiversity for conservation. Here we report the scientific discovery of a new primate species,
Distribution of
In this paper, we describe and name the new guenon species and discuss its relationship with its nearest congener and sister species,
The Congolese Wildlife Authority (Institut Congolais pour la Conservation de la Nature, ICCN) issued permits to the TL2 (Tshuapa, Lomami, and Lualaba) Project for all sites where biological samples and field observations were made. The ICCN is the governmental authority that has jurisdiction over the wildlife of this territory. Institutional Animal Care and Use Committees (IACUC) approval was not required for the noninvasive behavioral observations and biological samples of wild monkeys used in this study. IACUC protocols were followed for the collection of one skin snip specimen from a captive monkey. For the specimens collected from hunted animals, we obtained approval from the hunters to use these samples and no animal was hunted for the purpose of research. We acquired specimens only opportunistically in villages outside of the forest and we did not request samples from all lesula available to avoid targeting this species. When we encountered captive monkeys in villages, we photographed them with permission from the owner. We advised owners on the monkeys’ care and discouraged owners to acquire wild animals as captives. All the necessary exportation and importation permits were acquired by CITES, Centers for Disease Control and Prevention and the U.S. Fish and Wildlife Services.
Seven specimens of
All
Craniodental linear measurements were taken using digital calipers and recorded to the nearest tenth of a millimeter (
We digitized the landmarks on each cranium using a Microscribe G2X digitizer (Immersion Corp). The individual crania were immobilized in a bed of Play-Doh, and the landmarks were digitized in two sets–one superior view and one inferior view. All bilateral landmarks were digitized on the right side of the cranium. As reference points, five landmarks were digitized in both views. Subsequently, the two sets of landmarks were combined using the program DVLR v. 0.4.9
We assembled a ∼4.6 kb contig of X-chromosomal DNA from three overlapping amplicons for the newly surveyed
We amplified and sequenced a ∼2.2 kb segment of the Testis-Specific Protein, Y-chromosome (TSPY) using primers and protocols described by Tosi
Amplified products were cleaned with exonuclease I and shrimp alkaline phosphatase
We used a phylogenetic analysis in a comparative context to address the question of species vs subspecies status for the
The novel TSPY and Xq13.3 homolog sequences of
Maximum likelihood analyses using the appropriate models of molecular evolution were conducted first using PAUP* 4.0b10
Bayesian analyses of both datasets were conducted using MRBAYES 3.11
We used the BEAST 1.5.3 software package
The results of the four separate BEAST runs of both
We recorded vocalizations of
We made recordings with a Marantz PMD 660 digital recorder equipped with a Sennheiser condenser microphone module (K6/K6P). All recordings were digitized with Raven 1.3 (Cornell Laboratory of Ornithology, Ithaca, New York) using a 1024-point fast-Fourier transform, Hanning window function. We used the resulting spectrographic displays to extract direct measurements of 11 spectral and temporal parameters and five calculated derived measures (
Trained teams familiar with the calls of all the primates in the TL2 region conducted vocalization surveys between 13 March and 4 December 2009. Vocalization surveys were conducted at a total of 117 survey posts distributed systematically in six, 30 km×30 km blocks that had been previously selected for intensive large mammal inventories (18–22 vocalization surveys per block). The survey area included locations within
We collected data on
During survey periods, data were collected daily, from 6∶30 to 14∶00 by two experienced observers moving along the area’s permanent transect grid. Observers recorded the time and GPS waypoint for all primate groups encountered (seen and heard, or heard but not seen). For groups that were seen, we estimated the perpendicular distance from the estimated center of each to the transect line, counted the number of animals seen by species, and estimated the number of additional animals that could not be counted accurately. For groups that were heard but not seen, we recorded the occurrence of species based on vocalizations.
For each encounter with
The electronic version of this document does not represent a published work according to the International Code of Zoological Nomenclature (ICZN), and hence the nomenclatural acts contained in the electronic version are not available under that Code from the electronic edition. Therefore, a separate edition of this document was produced by a method that assures numerous identical and durable copies, and those copies were simultaneously obtainable (from the publication date noted on the first page of this article) for the purpose of providing a public and permanent scientific record, in accordance with Article 8.1 of the Code. The separate print-only edition is available on request from PLOS by sending a request to PLOS ONE, 1160 Battery Street, Suite 100, San Francisco, CA 94111, USA along with a check for $10 (to cover printing and postage) payable to “Public Library of Science”.
In addition, this published work and the nomenclatural acts it contains have been registered in ZooBank, the proposed online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix “
The scientific discovery of
Subsequent searches in Opala and in the Yawende area turned up other male and female captive juvenile lesula; all were photographed and some monitored for several months afterwards. Our first observation of the species in the wild was in the Obenge area (S 1.38461°, E 25.03749°) in December 2007 where the species is well known by local hunters.
YPM 14080, adult male, skin and skull (
Photographs show lateral (A) and anterior (B) views of the cranium and mandible, and occlusal views of the mandible (C) and cranium (D). Scale bar in each frame = 1 cm.
YPM 14189, subadult female, skin and skull, collected in the forest near Yawende, S 1.06571°, E 24.44838°, 450 m asl. YPM 14190, subadult female, skin and skull, collected near the Lomami River, S 1.42801°, E 25.01601°, 415 m asl. YPM 14191, adult male, skin and skull, collected near Obenge Village, S 1.38145°, E 25.03843°, 420 m asl. YPM 14192, subadult female, skin and skull, collected west of Obenge, S 1.40129°, E 24.97498°, 460 m asl. See
West Bank, Lomami River (S 1.02237° to S 1.4280°, E 24.42368° to E 25.03843°), Democratic Republic of Congo (
A mane of long grizzled blond hairs frames a protruding pale, naked face and muzzle, with a variably distinct cream colored (color 54
Left: Adult male, Yawende, DRC. Photograph by M. Emetshu. Right: Subadult female, Opala, DRC. Photograph by J. A. Hart.
Portraits: Captive adult male
Cranium with a distinctive elongated nasal profile and large orbits (
Comparative crania of
A medium sized, long-limbed monkey with a slender body. Naked facial skin, eyelids, and ear pinnae vary from pale pinkish gray to tannish brown. Buff diadem and vertical cream nose stripe are variably present. Chin, throat, and upper ventrum are yellowish buff, contrasting with black lower ventrum and abdomen. Skin and pelage on shoulder and forelimb are black. Hair over the entire dorsum is banded buff or amber with black. Typically, the base of the hair grades from white to gray to black and then exhibits 3–4 bands of buff/amber as one moves up the hair distally/dorsally. Anterior 2/3 of the dorsum, including head and mane, is buff, grading into amber over posterior third of dorsum and onto base of tail. Distinctively, there is a prominent amber median stripe on the distal half to one-third of the dorsum. The amber banding is restricted to the medial portion of the posterior third to half of the dorsum (see
Juveniles have a pale blond pelage overall, lightest on the throat and upper ventrum (
Sexual dimorphism is present for body and canine size, with the adult male larger. An adult male exhibits a bright blue scrotum and perineum (
Hunter-killed
Frequency (Hz) is on the Y-axis; time (sec) is on the X-axis. Mean values for acoustic measures for each species are indicated: 1. High frequency, 2. Low frequency, A. Start frequency, B. Q1 frequency, C. Q3 frequency, D. End frequency, dotted line: slopes.
Adult males (n = 2): head and body length 47–65 cm, weight 4.0–7.1 kg. Subadult females (n = 2): head and body length 40–42 cm, weight 3.5–4.0 kg. See
The specific name acknowledges the Lomami River near which the type specimen was found and which traces the eastern boundary of the species’ range.
Occurs in mature terra firma evergreen forests.
Lesula (Kingengele, Kilanga, Kimbole), Kifula (Kinyamituku), Tou (Kitetela).
Similarities in cranial, skin and pelage traits between
The cranium of
The variably prominent white to cream colored vertical nose strip shared by
The blue perineum, buttocks and scrotum displayed by adult males are comparable in size and coloration in
In both species, the juvenile is pale, uniformly colored and differs markedly from adults (
Overall, the olive-maned
The juvenile pelage of
The cranium of
Maximum likelihood and Bayesian phylogenetic analyses of TSPY sequence data clearly show that
Dashed gray lines highlight the inferred divergence date between
Molecular divergence date estimates of the most recent common ancestor (MRCA) of the
Values shown are mean and 95% Confidence Interval for estimates. See
The dawn boom chorus is the most conspicuous vocalization of both
The booms of
The calls differed significantly (Welch Two Sample t-test, n = 49, p<0.05) on two of 16 measured acoustic parameters: High frequency and 3rd Quartile Frequency Differences. Three other parameters (End Frequency, Start Frequency, and 1st Quartile Frequency) were nearly significant (range: p = 0.054–0.065) (
We found
Multiple individuals were recorded calling at 27 of 34 (79.4%) sites where
We found no evidence for seasonal variation in calling rates; however, we found marked differences in the occurrence of calling in different portions of the species known range. We recorded vocalizations at 11 of 40 (27.5%) listening points in the southern third of the range. In contrast, vocalizing animals were recorded at 16 of 19 listening points in the region of the Losekola Study Area and at 8 of 11 listening points in the northern portion of the species range. These latter two locations are in the Tutu River basin, a tributary of the Lomami River. These apparent differences in occurrence of
We observed 48 individual
Fourteen animals fled at or subsequent to encounter. Nine animals fled on the ground including three individuals that descended to the ground from the understory or middle level stratum to flee. Of the five animals fleeing in the strata above ground, only one climbed from the ground to flee. One animal first seen in the canopy fled laterally to a second canopy and then became stationary. In the other three observations, animals located above the ground fled in the same stratum in which they were first encountered.
Animals were recorded feeding or foraging on eight first encounters totaling 26 individuals. In five observations of feeding (20 animals total)
We made one exceptional observation of an apparent attack by a crowned eagle (
We applied a comprehensive approach to assess the evolutionary distinctiveness and appropriate taxonomic naming of the new primate discovery
Morphological and genetic analyses support the species designation of
Both
Element |
|
|
Known Range | 17,000 km2 | 180,000 km2 |
Elevation range | 400–615 m | 450–3500 m |
Habitat | Mature evergreen terra firma forests of the central Lomamiand upper Tshuapa basins including mixed andmonodominant forests. | Mature terra firma and secondary forests from lowlandformations in the west to montane forests and bambooin the Albertine Rift. |
Markings | Nose strip diffuse and off-white and blends into paleexposed face skin. Adult male: bright aquamarine scrotumand large perineal patch. Young male: pale gray, or faintlyblue scrotum and perineal patch. Female: pale gray perinealarea, sometimes with a bluish caste. | Nose stripe white, prominent, sharply demarcates the face.Nose stripe reduced or absent in some individuals (see |
Vocal behavior | Low frequency, descending ‘boom’, most frequent duringdawn chorus. Booms can be elicited by imitating eagle calls. | Low frequency, descending ‘boom’, most frequentduring dawn chorus. Booms not readily elicited by imitatingeagle calls. |
Olfactory signals | No information | Sternal (apocrine) glands and ritualized chest rubbingobserved in captives. |
Positional behavior | Ground to canopy. Feeds, moves and flees on ground. | Ground to canopy. Terrestriality varies by habitat, mostfrequent in montane habitats. |
Associations with other primates | Frequent member of multi species associations of primates. | Occasionally joins multi species associations in Ituri Forest.Forages terrestrially with duikers in Ituri. |
Group composition and size | Groups of adult females and their offspring and typicallyone adult male. Group size unknown, up to 5 individualsseen together. | Groups of adult females and their offspring and typicallyone adult male. Group size typically 1–15. One group of 22in Nyungwe Forest. One apparently temporary associationof ∼40 individuals seen in the Ituri Forest. |
Terrestrial vegetation in diet | Marantaceae important dietary component. Feeds onfallen fruit beneath arboreal primates. | Marantaceae and fungi reported in Ituri. Bamboo in montanehabitats. Feeds on fallen fruit beneath arboreal primates. |
Predators | Crowned eagles ( |
Leopard ( |
Speciation in African primates has been linked to Pleistocene forest refugia, riverine barriers, and transitional environments
Common name |
Scientific name |
Distribution within the TL2 region | Endemic to TL2 region | |
West Bank Lomami River | East Bank Lomami River to Congo–Lualaba River | |||
Bonobo |
|
Present | Present | No |
Tshuapa Red Colobus |
|
Present | Absent | No |
Lomami River Red Colobus |
|
Absent | Present | Yes |
Sclater’s Angola Colobus |
|
Present | Present | No |
Black Mangabey |
|
Present | Present | No |
Congo Basin Wolf’s Monkey |
|
Present | Present | No |
Kasuku River Wolf’s Monkey |
|
Absent | Present | Yes |
Lomami River Blue Monkey |
|
Present | Present | Yes |
Katanga Red-tailed Monkey |
|
Present | Present | No |
Lesula |
|
Present | Absent | Yes |
De Brazza’s Monkey |
|
Present | Present | No |
Taxonomy follows Grubb
At present, TL2 remains remote from human expansion, and there is no logging or mining. Hunting is the immediate threat to the faunas of TL2. We provide a provisional IUCN Red List Assessment of Vulnerable
The conservation of
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(AIF)
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JAH, TBH, ME and AV acknowledge the Insitut Congolais pour la Conservation de la Nature (ICCN). JAH, TBH, ME and AV thank the TL2 field teams, especially G. Paluku, K. Bafumoja, S. Dino, C. Kibambe, E. Mpaka, and O. Omene. KMD, ASB and AJT thank Dr. Todd Disotell for support to conduct genetic research at New York University’s Molecular Anthropology Laboratory. CCG and EJS thank Kristof Zyskowski and Gregory Watkins-Colwell in the Division of Vertebrate Zoology of the Yale Peabody Museum for their help with the YPM specimens from DRC and Eileen Westwig for access to specimens at the AMNH. We thank Matthew Shirley, Colin Groves, Christian Roos and an anonymous reviewer for their thorough review of the manuscript and comments. Special thanks to the editor Samuel Turvey for his assistance and helpful comments.