ISSN: 2167-6801

Journal of Primatology

The International Open Access

Journal of Primatology

Executive Editors

Robert J. Adams
Johns Hopkins University, USA

Bruce M. Rothschild
University of Kansas, USA

Agnes Lacreuse
University of Massachusetts, USA

Magdalena N Muchlinski
University of Kentucky, USA

Daniel D. Myers
University of Michigan Medical School, USA

Available online at: OMICS Publishing Group (www.omicsonline.org)

T his article was originally published in a journal published by

OMICS Publishing Group, and the attached copy is provided by
OMICS Publishing Group for the author’s benefit and for the benefit
of the author’s institution, for commercial/research/educational use
including without limitation use in instruction at your institution,
sending it to specific colleagues that you know, and providing a copy
to your institution’s administrator.
All other uses, reproduction and distribution, including without
limitation commercial reprints, selling or licensing copies or access,
or posting on open internet sites, your personal or institution’s
website or repository, are requested to cite properly.

Digital Object Identifier: http://dx.doi.org/10.4172/2167-6801.1000107

 Ruiz-García et al., J Primatol 2013, 2:1
http://dx.doi.org/10.4172/2167-6801.1000107

Primatology
Research
Research Article
Article Open
OpenAccess
Access

Molecular Genetics Analysis of mtDNA COII Gene Sequences Shows Illegal

Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia
M. Ruiz-García1*, C. Vásquez1, E. Camargo1, L. F. Castellanos-Mora2, H. Gálvez3, N. Leguizamón4 and J. M. Shostell5
1
LLaboratory of Molecular Population Genetics-Evolutionary Biology, Department of Biology, Pontificia Javeriana University, Cra 7 ª No 43-82, Bogota DC, Colombia
2
Omacha Foundation. Bogota DC, Colombia
3
Veterinary Institute of Tropical Research and Experiment Station H-(IVITA), Iquitos, Peru
4
Districtal Enviromental Secretary (SDA), Bogota, Colombia
5
Department of Biology, Penn State University-Fayette, Uniontown, PA 15401-0519, USA

Abstract
Night monkeys of the Aotus genus are employed by several laboratories in studies of malaria and other tropical
diseases. Some institutions, such as the Centre for Reproduction and Conservation of Non-Human Primates (CRCP,
IVITA) at Iquitos (Peru), breed exemplars of A. vociferans and A. nancymaae for laboratory purposes. However, as
several Colombian journals and newspapers have reported, an important laboratory in Colombia (FIDIC) employs
illegally imported individuals of the species, A. nancymaae, which is not part of the native primate fauna of this
country. Our molecular analysis using mtDNA COII gene sequences demonstrates that the FIDIC’s laboratory
contains A. nancymaae specimens which could have been acquired through illegal trafficking of this species. This
laboratory does not possess a legal license to obtain and import specimens of this Aotus species from Peru or Brazil,
where this species is native.

Keywords: Aotus; mtDNA COII gene sequences; Phylogenetic
analyses; Illegal trade; Colombia; Peru
Introduction
Aotus species are the unique nocturnal monkeys in Latin America.
The systematics of Aotus species is complex [1-4] as is the relationships
of the genus Aotus with other Neotropical primate genera [5-9].
These species have been employed in the search for vaccines against
the malaria and other tropical diseases. However, not all of the Aotus
species are useful for this task. This is due to the fact that different Aotus
taxa have different susceptibility to the malaria parasite. Schmidt [10]
demonstrated that A. nancymaae was highly or completely resistant
to nine strains of Plasmodium falciparum but not to two strains of
Plasmodium vivax, while A. griseimembra was extremely susceptible
to these nine P. falciparum strains as well as to the two strains of P.
vivax. In the same analysis, Schmidt [11] showed that the individuals
of A. nancymaae quickly recovered, or did not present any evidence
of infection, with Palo Alto, Malayan, Camp Sadun and Camp-CH/Q
P. falciparum strains, whereas the infections were fulminating in A.
griseimembra.
In related research, when working with A. nancymaae. Collins
et al. [12] showed that there was some possibility of immunity to P.
falciparum (Panamá II strain) that is related with blood sera with
three alpha-globulins. However, A. griseimembra only has two alpha-
globulins and the same P. falciparum strains were highly infectious
in this species. Also, Taylor and Siddiqui [13] demonstrated that
the individuals of A. griseimembra died when infected with African
strains (Uganda-Palo Alto) of P. falciparum, while A. zonalis and A.
nancymaae survived these infections. In contrast, bisalbuminemia was
shown to be present in A. nancymaae but not in A. griseimembra [14].
For these reasons, A. nancymaae has been used as a biomedical model
for vaccines against malaria, such as work by Gozalo et al. [15] who
employed this species to probe the 19 Kd MSP-1 terminal falciparum
vaccines.
Therefore, A. nancymaae has several medical interesting traits that
other Aotus taxa do not possess that could be helpful to those laboratories
that have an anti-malarial focus. These attributes are reflected in the
publications resulting from the research conducted at the Fundación
Inmunológica de Colombia (FIDIC) in Leticia (Colombian Amazon),
which indicates that, indeed, both A. nancymaae and A. nigriceps were
used in the construction of a malaria vaccine [16-20].
However, both A. nancymaee and A. nigriceps are species that are
not part of the indigenous primate fauna of Colombia [1,21] This fact
suggests that these Aotus species were illegally imported to Colombia
by FIDIC for research on malarial vaccines and therapies. In fact,
FIDIC has been continuously accused of trafficking Aotus from other
countries without any permission and control.
More than 20 years ago, this laboratory created the first synthetic
vaccine against malaria (spf66). Although its application in Colombia
and Tanzania only reached 40% efficacy and the production of the
vaccine has now been discontinued, its creation provided benefits to
the chief of this laboratory, who has obtained important recognitions
(e.g. the Principe de Asturias award in Spain, 1994).
Corpoamazonía (Corporación para el Desarrollo Sostenible del
Sur de la Amazonía), the governmental environmental authority in the
Colombian Amazon, conceded a legal license to FIDIC for trapping
1,600 A. vociferans during an eight month period that began on
*Corresponding author: M. Ruiz-García, Laboratorio de Genética de
Poblaciones, Molecular-Biología Evolutiva Departamento de Biología,
Pontificia Universidad Javeriana, Cra 7ª No 43-82, Bogotá D.C., Colombia,
E-mail: mruiz@javeriana.edu.co, mruizgar@yahoo.es
Received September 14, 2012; Accepted October 14, 2012; Published October
16, 2012
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez
H, et al. (2013) Molecular Genetics Analysis of mtDNA COII Gene Sequences
Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J
Primatol 2:107. doi:10.4172/2167-6801.1000107
Copyright: © 2013 Ruiz-García M, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited.

J Primatol
ISSN: 2167-6801 JPMT, an open access journal Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107
September 13, 2006, in areas close to San Juan de Atacuarí (an Indian
community at the border of Colombia and Peru at the Amazon River)
and in the Colombian islands between Leticia and Puerto Nariño on
the Amazon River. This permission was renewed at least one additional
time. However, in the last months of 2008, Corpoamazonía opened an
inquiry (No. 000102) of the FIDIC because there were accusations and
evidence of illegal Aotus trade from different points of the Peruvian and
Brazilian Amazon to Leticia. Some of this evidence was published by
the Colombian journal “Cambio” and is summarized below.
To begin with, a Corpoamazonía functionary named Claudia Marín
visited the Aotus collection retained in the FIDIC, and she affirmed
that the documentation for the collection was incomplete and that a
fraction of animals imported from Peru and Brazil, recorded in the
Aotus reception book, were obtained without any legal authorization.
Similarly, journalists of “Cambio” read through the entrance reception
books of 2004 and 2005, and stated that a fraction of the animals from
the FIDIC collection were illegally obtained from Peru and Brazil. This
news was troubling from a biological perspective because the Aotus
populations from the southern bank of the Amazon River in Peru and
Brazil belong to a Aotus species that is different from A. vociferans,
which is the species present at the Colombian Amazon [21,22] A.
nancymaae.
Besides this evidence, several people have explicitly declared that
they provided Aotus individuals to the FIDIC. For instance, some
Peruvian Ticuna Indians, like Mr. Víctor González, declared that, at
the beginning of October 2008, they transported about 120 Peruvian
Aotus animals captured at the southern Amazon bank from Puerto
Alegria (Peru) to Leticia (Colombia), a trip that took five hours (in
“peque peque”). They received 600,000 Colombian pesos (about 300
US dollars) and were asked to transport the animals during the first
hours of the day in the darkness, so they could avoid detection with
the consequence of being held in prison for three years. They were
further told that, if intercepted by the police or by the army, they
should declare that the animals were captured on the Colombian
side of the Amazon River. Similarly, inhabitants from other Peruvian
Amazon localities, such as Chinería, Barranquilla, Gamboa, Yahuma,
San José de Yanayacu or San Miguel, as well as those from Brazilian
Amazon localities, as Bom Sitio, Tucano, Sacambú or Puerto Brasil,
also declared that they illegally transported Aotus individuals to Leticia.
Evin Santos and Yolvin Santajaya, from the Barranquilla population
in the Peruvian Amazon, stated that they captured Aotus within the
past four years and that the FIDIC paid them 50,000 Colombian pesos
(about 25 US $) for each animal.
Unfortunately, some Brazilian and Peruvian authorities indicated
that they knew of this situation but could not prevent illegal Aotus
trafficking because they did not receive any cooperation from the
Colombian authorities. For example, Mr. Jaime Santander from the
Peruvian Instituto Nacional de Recursos Naturales (INRENA) stated
that he knows that there is illegal trade of Aotus from Peru to Colombia.
Not surprisingly, the chief of FIDIC rejected all of these accusations
(e.g., when he was questioned in a medical meeting in Asturias, Spain,
in May of 2009, information that was provided by TVE, Public Spanish
Television affirmed that they have only caught A. vociferans individuals
from the Colombian side of the Amazon River).
At the end of 2008, Corpoamazonía asked us if we could analyze a
fraction of the Aotus’s collection at FIDIC by using molecular markers
to identify the species that they represented. As we have some expertise
in molecular analyses with Neotropical primates [23-31] and other
wild mammals [32-38], we accepted its proposal. Corpoamazonía
J Primatol
ISSN: 2167-6801 JPMT, an open access journal
Page 2 of 9
provided us with samples from 24 individuals, randomly chosen by
Corpoamazonía, although we cannot observe the remainder 480
animals that were present in the FIDIC at that moment.
Once the samples were obtained, we sequenced the mitochondrial
DNA (mtDNA) COII gene in all of them. The MtCOII is an
indispensable gene that plays a key role in the terminal oxidative step of
energy metabolism by catalyzing the transfer of electrons from reduced
cytochrome c to oxygen to give water. It is also related to the synthesis
of ATP in oxidative phosphorylation [39]. This gene nearly shows a
two-fold increase in the rate of amino acid replacement in monkeys
and apes in contrast to the strepsirrhines. The important amino
acids encoded by this gene are usually conserved in primates, but the
acceleration found in haplorrhine species suggests increased variation
in the amino terminal end of the protein [40]. For instance, the extreme
carboxy-terminal end of COII protein is highly variable in Neotropical
primates, and shows increased additions and deletions of amino acids
relative to other mammals [40].
Previously, this mitochondrial gene has been used to infer
phylogenetic relationships in Primates (within the Hominoidea,
[41,42], within the Cercopithecoidea, [43], within the Strepsirrhi, [40]
and within several Neotropical primate genera: Aotus [2-4]; Alouatta
[44]; Ateles [31,45-47]; Cebus albifrons [30,31,34] Cebus capucinus
and Lagothrix [34]; Saguinus and other Callitrichidae [34,48,49].
Nevertheless this gene, like other mitochondrial loci, involves several
problems for primate phylogeny (heterogeneity in base composition at
each codon position, different transition and transversion substitution
rates and third-codon position saturation). However, [48] showed that
at intrageneric and intraspecific levels, the sequences of this gene are
phylogenetically informative, although not at a higher taxonomic level.
Thus, the main aim of the current work was to determine if, within
the FIDIC Aotus collection, there were samples from A. nancymaae
which could support the continuous accusations of illegal trade made
by many people against this institution.
Material and Methods
Samples
We analyzed a total of 111 Aotus individuals for 715 base pairs
(bp) of the mtDNA COII gene. These samples were as follows: Twenty-
four (24) samples came from the FIDIC (Leticia), 12 samples of A.
vociferans originated from three different locations (six samples from
Santa María and Tamboryacu Rivers, which are tributaries of the Napo
River and the Nanay River at the Loreto Department in the Peruvian
Amazon; five Colombian individuals from Puerto Nariño to San Juan
de Atacuarí in the Colombian Amazon; and one individual from Lago
Agrio at the Ecuadorian Amazon), 27 individuals of A. nancymaae
sampled in different points at the Peruvian Amazon (Tahuayo River,
Quebrada Yanayacu, Requena-Tapiche River, Pucallpa-Ucayali River,
Huallaga River and Caballococha-Amazon River), 19 samples of A.
griseimembra from the Córdoba, Sucre and Atlantico Departments
in northern Colombia, nine samples of A. brumbacki from the Meta
Department in Colombia, one sample of the recent species described
as A. jorgehernandezi [27] from the Bremen Reserve at the Quindio
Department (Colombia), three samples from the eastern Andes
cordillera in Colombia and classified as A. lemurinus, two samples of
A. nigriceps (one from Tarapoto at the San Martín Department, and
the other obtained in Yarinacocha at the Ucayali Department, both
in the Peruvian Amazon), two samples of the rare species A. miconax
(one from an animal confiscated in Chiclayo and other sampled at
Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107
Bongaro, northern Peru), four samples of A. azarae boliviensis (two
from San Javier, Beni Department and two from the Santa Cruz
Department, both in Bolivia) and three samples of A. azarae azarae
from west of Paraguay River in Paraguay. Six other samples which were
not “a priori” taxonomically identified and that we named “problem”,
were also sequenced. In the Results and the Discussion sections, we
will classified these samples and comment on them because these
are important for understanding the illegal commerce of Aotus in
Colombia. Additionally, tree specimens of Cebus capucinus (from
the Cauca and Córdoba Departments in northern Colombia) were
employed as out-groups. In addition, some trees were obtained without
out-groups following [50] in order to accommodate recently debated
issues about molecular dating of recent phylogenetic splits.
Molecular analyses
For the animals from FIDIC, one cm3 of blood was obtained
and preserved in disodium EDTA. For the other animals, which
were caught as pets by Indian communities, we extracted DNA
from hair (with follicles). We obtained blood DNA using phenol-
chloroform [50] and hair DNA using 10% Chelex resin [51]. We used
the primers L6955 (5’-AACCATTTCATAACTTTGTCAA-3’) and
H7766 (5’-CTCTTAATCTTTAACTTAAAAG-3’) to PCR amplify the
mtCOII gene [2,45]. We performed each PCR in a 50-μl volume with
reaction mixtures including 4 μl of 10x Buffer, 6 μl of 3 mM MgCl2, 2
μl of 1 mM dNTPs, 2 μl (8 pmol) of each primer, 2 units of Taq DNA
polymerase, 13.5 μl of H20 and 2 μl of DNA. PCR amplifications were
carried out in a Geneamp PCR system 9600 (Perkin Elmer) and in a
Bio-Rad thermocycler. We used the following temperatures and cycles:
95°C for 5 minutes, 35 cycles of 45 s at 95°C, 30 s at 50°C and 30 s
at 72°C and a final extension time for 5 minutes at 72°C. Using the
molecular weight marker φX174 DNA digested with Hind III and Hinf
I, we checked all the amplifications, including positive and negative
controls, in 2% agarose gels. We purified the amplified samples with
membrane binding spin columns (Qiagen), directly sequenced the
double-stranded DNA in an ABI 377A automated DNA sequencer in
both directions and then repeated the sequencing of each sample to
ensure accuracy.
The use of the mtCOII gene for inferring primate phylogeny
can be problematic because of the rapid rate of molecular evolution
at mitochondrial loci and the saturation problem regarding a
phylogenetically informative signal at the 3rd position within codons
at the inter-generic level. Nevertheless, [48] have shown that this gene
can be informative at the intra-generic level for New World primate
taxa. However, we cannot completely exclude the possibility that some
sequences obtained in this study represent numts (mitochondrial DNA
fragments inserted into the nuclear genome) rather than true mtDNA
[52]. However, we note that all amino acid translations of the sequences
obtained showed the presence of initial start and terminal stop codons
and the absence of premature stop codons. Therefore, there is high
confidence that the sequences analyzed are not numts.
Population genetics analysis
Following Ashley & Vaughn [2], we used a transition-transversion
rate ranging from 5:1 to 15:1 to obtain diverse phylogenetic trees. We
used the methods of genetic distances (with the neighbour-joining
algorithm; [53], maximum likelihood, and maximum parsimony (with
close-neighbour-interchange with search level 1 and random addition
with 100 replicates) using Mega 5.0 and PAUP*4.0b8. A total of 1000
bootstraps were run to determine the consistence of the different nodes
obtained in the trees.
J Primatol
ISSN: 2167-6801 JPMT, an open access journal
Page 3 of 9
Results
The trees obtained with genetic distances (with, or without,
heterogeneous evolutionary patterns and lineages or with, or without,
different rates among sites), with maximum likelihood and with
maximum parsimony procedures, provided identical results for the
FIDIC individuals and were only slightly different from the bootstrap
results. Herein, we show the neighbour-joining tree with the Tamura
and Nei’s genetic distance without out-groups and the consensus
maximum parsimony tree (Figures 1 and 2). Two ensembles contained
sequences from A. vociferans. Twenty-two of the FIDIC samples
clustered with one of these A. vociferans’ ensembles (08-496, 08-323,
08-243, 08-308, 08-098, 08-051, 08-299, 08-509, 08-512, 08-058, 08-
280, 08-515, 08-268, 08-32311, 08-046, 08-054, 08-053, 08-285, 08-424,
08-221, 08-092 and 08-485). Therefore, it seems clear that 22 out 24
individuals sampled by the FIDIC can be taxonomically identified as
belonging to A. vociferans, consistent with the permission that this
institution has used to obtain individuals from this species for medical
research. However, two other individuals (numbers 08-575 and 08-246)
clearly clustered with other A. nancymaee individuals. This observation
supports claims of illegal traffic of several species of Aotus on the part
of the FIDIC.
A second relevant question with regard to illegal trafficking was
the inclusion of the six individuals named “problem” in the cluster
basically constituted by individuals of A. nancymaae. These animals
were sampled between Puerto Nariño and San Juan de Atacuarí in
the Colombian Amazon in 2009 and [1] and others argue that A.
nancymaae inhabits the southern side of the Amazon River (Peru and
Brazil) and not the northern bank of the Amazon River in Colombia.
Thus, the presence of these A. nancymaee in the Colombian side is
seemingly anomalous, and these details will be discussed below.
From a phylogenetic perspective, it is clear that the northern Amazon
River Aotus formed a monophyletic cluster, although differently to that
found by [4] with a smaller Aotus sample, the supposed different species
did not show haplotype monophyly within this group. Together the
vociferans haplotypes, there are some haplotypes of A. griseimembra,
A. brumbacki and that of A. jorgehernandezi. The haplotype of the
latter taxon was identical to a haplotype found in A. brumbacki. The
haplotypes of A. lemurinus were very similar to several haplotypes of
A. brumbacki, while both types of haplotypes for A. lemurinus and A.
brumbacki were highly related to the main A. griseimembra cluster.
The southern Amazon River Aotus group is a polyphyletic, as shown
by [4]. This southern group was considerably more heterogeneous than
the northern group. In this analysis, A. nigriceps was more related to
the northern Aotus group than to the other southern Aotus group. The
two haplotypes of A. miconax were polyphyletic and were enclosed
within the A. nancymaee cluster. This observation could be support for
the argument that A. miconax is a very recent taxon (species). That is,
A. miconax could be formed from ancestral individuals or populations
of A. nancymaee who became adapted to live at higher altitudes in the
Andes.
The unique considerable difference between both trees showed
was in reference to A. azarae boliviensis. For the distance tree, this
population formed a monophyletic group inside one A. nancymaee
cluster. This could suggest that A. azarae boliviensis descended from
an ancestor related with A. nancymaee. However, the maximum
parsimony tree showed A. azarae boliviensis as the most divergent
Aotus taxa was analyzed. In whatever case, it seems clear that A. azarae
Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107

Page 4 of 9

FIDIC8-299
FIDIC8-323-11
FIDIC8-512
FIDIC8-051
FIDIC8-509
27
FIDIC8-515
FIDIC8-054
FIDIC8-268
30
FIDIC8-058
FIDIC8-053
FIDIC8-280
18 FIDIC8-046
FIDIC8-221
5
FIDIC8-424
Aotus vociferans 171 D Colombia
26 Aotus vociferans 38 Colombia
80 Aotus vociferans 4 Peru
4122 Aotus vociferans 10 Peru
11 FIDIC8-285

63 FIDIC8-092
49 FIDIC8-485

FIDIC8-323
46
FIDIC8-308
FIDIC8-098
37
FIDIC8-243
FIDIC8-496
Aotus brumbacki 16 Colombia
78
Aotus brumbacki 1950 Colombia
82
Aotus brumbacki 127 Colombia
63
Aotus jorgehernandezi Quindio Colombia
Aotus vociferans 160 Ecuador
46 Aotus vociferans 13 Colombia
22 Aotus vociferans 5 Peru
40
Aotus griseimembra 42 Sucre Colombia
Aotus brumbacki 176 Colombia
29
Aotus vociferans 14 Colombia
Aotus vociferans 1 Peru
19
Aotus vociferans 12 Peru
Aotus vociferans 57 Peru
19
79 Aotus vociferans 43 Colombia
Aotus griseimembra 350 Colombia
Aotus griseimembra 50 Coloso Colombia
53 Aotus lemurinus Victoria Caldas Colombia
26 Aotus lemurinus 9 Colombia

Aotus lemurinus 87 Colombia

Aotus brumbacki 125 Colombia
36
Aotus brumbacki 14 Colombia
Aotus brumbacki 18 Colombia
96 Aotus brumbacki 13 Colombia

Aotus brumbacki Meta Colombia

Aotus griseimembra Vicente Chucuri Colombia
67 Aotus griseimembra Monteria Colombia
95
Aotus griseimembra 35 Colombia
71
54 Aotus griseimembra 68 Colombia
Aotus griseimembra 103 Colombia
80
Aotus griseimembra 21Colombia
Aotus griseimembra 19 Colombia
37 Aotus griseimembra 6260 Colombia

Aotus griseimembra 4989 Colombia

Aotus griseimembra 6 Colombia
39 Aotus griseimembra 10 Colombia

Aotus griseimembra 26 Colombia

86
Aotus griseimembra 205 Colombia
Aotus griseimembra 20 Colombia
Aotus griseimembra 11 Colombia
64
Aotus griseimembra 12 Colombia
Aotus nigriceps Yarinacocha Peru
100 Aotus nigriceps Tarapoto Peru
Aotus nancymaae 680 Peru
Aotus nancymaae 1 Peru
Aotus nancymaae 16 Peru
69
Aotus nancymaae 71 Peru
58
Aotus nancymaae 6 Peru
Aotus nancymaae 126 Peru
Aotus nancymaae 21 Peru
47 Aotus nancymaae 15 Pucallpa Peru
13
Aotus nancymaae 68 Peru
Aotus nancymaae Loreto Yacu Peru
Aotus nancymaae 121 D Peru
Aotus nancymaae 5 Peru
6 Aotus nancymaae Miconax Bongara Peru

67 Aotus nancymaae 21 Miconax Chiclayo Peru

Aotus nancymaae 54 Peru
100 Aotus nancymaae Huayaga Peru
Aotus nancymaae 18 Peru
1
Aotus nancymaae 78 Peru
Aotus nancymaae 11 Peru
72
12 Aotus nancymaae 37 PROBLEM
FIDIC8-575
Aotus nancymaae 21 Peru 39 PROBLEM
54 Aotus nancymaae 9 Peru
77
Aotus nancymaae 42 PROBLEM
70 Aotus nancymaae 36 PROBLEM
Aotus nancymaae 41 PROBLEM
28
Aotus nancymaae 74 Peru
Aotus nancymaae Requena Peru
14 Aotus nancymaae 38 PROBLEM

Aotus nancymaae 8 Peru

FIDIC8-246
39 Aotus
nancymaae 19 Peru
Aotus
nancymaae 20 Peru
Aotus
nancymaae 76 Peru
Aotus
nancymaae 72 Peru
Aotus
nancymaae 80 Peru
Aotus azarae azarae 21 Paraguay
100 Aotus azarae azarae 55 Paraguay
100 Aotus azarae azarae 56 Paraguay
Aotus azarae boliviensis 151 Bolivia
Aotus azarae boliviensis 259 Bolivia
100 Aotus azarae boliviensis 248 Bolivia
100 Aotus azarae boliviensis 149 Bolivia

0.05

Figure 1: Neighbor-joining tree with the Tamura-Nei genetic distance applied to the 111 Aotus analyzed for the mtCOII gene sequences. The animals with numbers are
those analyzed from the FIDIC collection. Three Cebus capucinus sequences were employed as out-groups. The numbers in the nodes are the bootstrap percentages.
In different colors, the different species of Aotus analyzed. The six animals named “problem” were Aotus nancymaee in Colombian territory. The arrows indicate illegal
traffic of Aotus in Colombia.

J Primatol
ISSN: 2167-6801 JPMT, an open access journal Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107

Page 5 of 9

FIDIC8-058
FIDIC8-268
FIDIC8-323-11
FIDIC8-299
FIDIC8-280
FIDIC8-046
FIDIC8-509
FIDIC8-054
FIDIC8-512
FIDIC8-424
100 65
Aotus vociferans 171D Colombia
66
Aotus vociferans 38 Colombia
FIDIC8-221

100 Aotus vociferans 4 Peru

67 Aotus vociferans 10 Peru
FIDIC8-285
FIDIC8-092
66 FIDIC8-485
100
FIDIC8-051
FIDIC8-053
FIDIC8-515

65 Aotus vociferans 4 Peru

Aotus jorgehernandezi 4 Quindio Colombia
100
Aotus brumbacki 16 Colombia
Aotus brumbacki 127 Colombia
100 FIDIC8-496
FIDIC8-098
65
FIDIC8-243
FIDIC8-323
FIDIC8-308
Aotus vociferans 160 Ecuador Colombia
Aotus brumbacki 176 Colombia
Aotus griseimembra 350 Colombia
Aotus vociferans 12 Peru
100
100 Aotus vociferans 13 Colombia
Aotus vociferans 5 Peru
Aotus griseimembra 4 Sucre Colombia
66
Aotus vociferans 14 Colombia
99
Aotus vociferans 43 Colombia
Aotus griseimembra 50 Coloso Colombia
Aotus vociferans 1 Peru
Aotus vociferans 57 Peru
66 Aotus brumbacki 14 Colombia
66 Aotus brumbacki 18 Colombia
Aotus brumbacki 125 Colombia
Aotus brumbacki Meta Colombia
100
Aotus brumbacki 13 Colombia
Aotus lemurinus 87 Colombia
100 Aotus lemurinus Victoria Caldas Colombia
100 Aotus lemurinus 9 Colombia
Aotus griseimembra Vicente Chucuri 50 Colombia
100 Aotus griseimembra Monteria Colombia
67 100
Aotus griseimembra 35 Colombia
Aotus griseimembra 10 Colombia
Aotus griseimembra 19 Colombia
Aotus griseimembra 21 Colombia
100 Aotus griseimembra 6 Colombia
98 Aotus griseimembra 20 Colombia
100 Aotus griseimembra 12 Colombia
Aotus griseimembra 11 Colombia
100
Aotus griseimembra 4989 Colombia
100 Aotus griseimembra 68 Colombia
Aotus griseimembra 103 Colombia
Aotus griseimembra 6260 Colombia
Aotus griseimembra 205 Colombia
Aotus griseimembra 26 Colombia
Aotus nigriceps Yarinacocha Peru
100 Aotus nigriceps Tarapoto Peru
Aotus nancymaae 76 Peru
100
Aotus namcymaae 80 Peru
FIDIC8-246
Aotus nancymaae 19 Peru
65 Aotus nancymaae 8 Peru
Aotus nancymaae 38 PROBLEM
Aotus nancymaae 74 Peru
100 Aotus nancymaae 20 Peru
Aotus nancymaae 72 Peru
Aotus nancymaae 9 Peru
100 Aotus nancymaae Requena Peru
66 Aotus nancymaae 11 Peru
100 Aotus nancymaae 37 PROBLEM
FIDIC8-575
65 Aotus nancymaae 42 PROBLEM
65
Aotus nancymaae 39 PROBLEM
65 Aotus nancymaae 36 PROBLEM
100 Aotus nancymaae 41 PROBLEM
66
100 Aotus miconax Chiclayo Peru
Aotus nancymaae 54 Peru
65 Aotus nancymaae 18 Peru
100
66 Aotus nancymaae 78 Peru
Aotus nancymaae Huayaga Peru
100 Aotus azarae boliviensis 259 Bolivia
100 Aotus azarae boliviensis 248 Bolivia
100
65 Aotus azarae boliviensis 149 Bolivia
66
Aotus azarae boliviensis 151 Bolivia
66
Aotus nancymaae 680 Peru
Aotus nancymaae 1 Peru
Aotus nancymaae 126 Peru
Aotus nancymaae 54 Loreta Yaku Peru
66 Aotus nancymaae 121D Peru
Aotus nancymaae 68 Peru
65 Aotus nancymaae 16 Peru
100 Aotus nancymaae 71 Peru
65
Aotus nancymaae 6 Peru
Aotus nancymaae 21 Peru
Aotus nancymaae 5 Peru
Aotus miconax Bongara Peru
Aotus nancymaae 15 Pucallpa Peru
Aotus azarae azarae 248 Paraguay
100 Aotus azarae azarae 55 Paraguay
100 Aotus azarae azarae 56 Paraguay
Cebus capucinus Costa Rica 172
Cebus capucinus Costa Rica 173
Cebus capucinus Costa Rica 174

Figure 2: Consensus maximum parsimony tree applied to the 111 Aotus analyzed for the mtCOII gene sequences. The animals with numbers are those analyzed
from the FIDIC collection. The numbers in the nodes are the bootstrap percentages. In different colors, the different species of Aotus analyzed. The six animals named
“problem” were Aotus nancymaee in Colombian territory. The arrows indicate illegal traffic of Aotus in Colombia.

J Primatol
ISSN: 2167-6801 JPMT, an open access journal Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107
azarae and A. azarae boliviensis should be not treated as subspecies of
the same species.
Discussion
Evidence of illegal trade of Aotus in Colombia
The presence of diverse Aotus taxa in Colombia makes this country
a fundamentally important area for our understanding of the evolution
of the genus. We can enumerate in Colombia at least the following
taxa: A. lemurinus, A. griseimembra, A. zonalis, A. brumbacki, A.
jorgehernandenzi, A. vociferans and probably A. trivirgatus, following
[21,22]. Furthermore, Colombia houses some important laboratories
that focus on the study of malaria and other diseases and that are
interested in Aotus species because of their differing susceptibilities to
malaria.
We now have molecular evidence that supports the illegal traffic of
Aotus in Colombia as related to the FIDIC. Two of the 24 individual’s
analyzed proceeding from the FIDIC presented sequences typically
from A. nancymaae (numbers 08-575 and 08-246) related to those
animals sampled at the Quebrada Yanayacu in the Peruvian Amazon.
Hershkovitz [1] identified A. nancymaae from specimens collected
from the north bank of the Samiria River above Estación Pithecia
within the Pacaya-Samiria National Park, at the Loreto Department
in Peru. This taxa has a karyotype of 2n=54, with nine metacentric,
two submetacentric and 15 acrocentric chromosome pairs. Following
to Ford (1994), most skull and upper facial dental measurements are
very large with regard to other nearby Aotus taxa, whereas lower dental
measurements were small. The geographical distribution of this species
is in the southern bank of the Amazon and Marañón rivers extending
up to the Ucayali River south of 7°S. In this southern boundary, some
hybridization with A. nigriceps could occur, as Hershkovitz (1983)
noted that some exemplars he obtained appeared to be intermediate,
phenotypically speaking, between A. nancymaae and A. nigriceps.
Groves (2001) mentioned a possible southern boundary with A.
nigriceps somewhere north of the Juruá River. Similarly, the presence
of A. nancymaae in the Contamana area neighbouring the Brazilian
Acre frontier was also mentioned where, within the Serra do Divisor
National Park [54], the presence of A. nigriceps was established by
Lopes and Rehg [55-59]. An A. nigriceps enclave could also occur in
the area of the Reserva Comunal Tamshiyacu-Tahuayo in the middle
of A. nancymaae’s territory [60]. In the west-to-east dimension, the
distribution of this species extends from the east bank of the Huallaga
River, possibly including the lower areas of the Peruvian eastern Andes
[61], to the Jandiatuba River in the Brazilian Amazon. One population
is present in a tiny region found north of the Amazon River between
the lower Tigre (and Tigrillo) and Pastaza Rivers (tributary of the
Marañón River). Thus, this taxon is basically a Peruvian species with
some distribution on the Yavarí and the Jandiatuba rivers in Brazil [62].
Accordingly, no study has mentioned this species as being present
in Colombia [21,22,54,63]. Thus, the presence of A. nancymaae in the
FIDIC Aotus collection agrees quite well with the accusations of illegal
trafficking against this institution in the last decades. Therefore, the use
of mtDNA sequences has sufficient discriminative power to detect the
illegal trade of Aotus species.
Other investigations have shown the existence of this illegal trade.
Torres et al. [63] studied 35 Aotus individuals for their karyotypes, and
noted that, of these 35 animals, five obtained from Leticia (Colombian
Amazon) were characterized by 2n=54, thus, were classified as A.
nancymaae. However, it is now clear that this species is not present
J Primatol
ISSN: 2167-6801 JPMT, an open access journal
Page 6 of 9
inside Colombia as an indigenous species. Four of these animals had the
Central Bioterio of the National Health Institute of Colombia as their
final destination. Another dead exemplar was deposited in the Natural
Sciences Institute of the Colombian National University (ICN 14019).
These animals were legally transported from Leticia to Bogota, but they
were taken illegally to Leticia from Peru and Brazil. Indeed, as [21]
affirmed, the idea of A. nancymaae being part of the Colombian fauna
is fallacious.
Previously, Ma et al. [64] analyzed 21 exemplars of A. nancymaae
(2n=54) that came from Tabatinga, the Brazilian village separated by a
street from the Colombian Leticia. Hershkovitz [1] later demonstrated
that these animals were obtained in the southern Amazon bank
opposite to Tabatinga in Brazil or in Peru. Most of these animals came
from the Estirao do Ecuador in the Yavarí River (Brazil). Hershkovitz
[1] clarified that the night monkey of the north Amazon bank, with
type locality in Tabatinga, belongs to the species A. vociferans.
A second direct investigation comes from a craniometric Aotus
study that the first author made in 2002 with material deposited in the
Mammalian Museum of the Humboldt’s Institute at Villa de Leyva
in Colombia. This author measured different cranial and dental traits
from 60 Aotus skulls. Interestingly, seven of these 60 exemplars were
donated by the chief of the FIDIC: Specimen 5223 (Aotus nancymaae,
male, Yahumá, Peru, 1984); specimen 5224 (A. nancymaae, female,
1983); specimen 5230 (A. nancymaae, female, Aramazá, Brazil, 1983);
specimen 5231 (A. nancymaae, male, Yahumá, Peru, 1983); specimen
5222 (A. nancymaae, female, Javarí River, Brazil, 1983); specimen 5227
(A. nancymaae, male, Aramazá, Brazil, 1983); and specimen 5228 (A.
nancymaae, male, Yahumá, Peru, 1983). These data suggest that the
illegal importation of A. nancymaae to Colombia has been occurring
for more than 25 years.
A third investigation comes from a shared personal experience of
two of the authors of this paper (M. R-G, and H. G). In January 2003,
these authors returned from an Amazonian expedition to capture pink
river dolphins for a population genetics and phylogeography project.
It was during this expedition that they came across 25 Peruvian
Aotus specimens released in Leticia. H. G., a veterinarian from the
Iquitos Primate Center, unambiguously classified all of the animals as
belonging to A. nancymaae and returned them to Peru. Who could be
interested in importing a large quantity of A. nancymaae to Leticia,
Colombia?
Between 2001 and 2003, the first author sampled Aotus in the area
from Leticia to San Juan de Atacuarí, including “Los Kilómetros”, El
Progreso, Santa Sofia, Zaragoza, El Vergel, Macedonia, Palmeras,
Puerto Nariño, Veinte de Julio, Tarapoto and other little communities
at the Loreto-Yaku River. Ten animals were sampled and all were
molecularly and phenotypically classified as being A. vociferans.
Curiously, the first and the fourth authors returned to the same area
in 2007-08 and again sampled night monkeys, in this case, four Aotus.
In this case, the analyses carried out in our laboratory showed that
the animals were molecularly A. nancymaae. More recently (2011),
two students of the Tolima University (Ana Silvia Diaz and Fernanda
Hernández) sent us hair samples from seven Aotus captured in the
same area. Six of the samples yielded DNA and were found to belong
to A. nancymaae. These are the six Aotus individuals named “problem”
in the trees shown in figures 1 and 2. Additionally, all of the Aotus
that these students observed in that area were A. nancymaae, not A.
vociferans, the species that we observed at the beginning of 2000. Thus,
it appears that the FIDIC liberates Aotus individuals after they have
been involved in experimental events. Furthermore, the liberation
Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107
area is the same region where we observed this radical change from A.
vociferans to A. nancymaee in the last decade. It is now clear that the A.
nancymaae illegally acquired by FIDIC could have been released into
the Colombian Amazon territory, an area where this species did not
originate and where an original unique species A. vociferans inhabited.
Both of these Amazonian Aotus species differ in their natural
history traits. The Centre for Reproduction and Conservation of
Non-Human Primates (CRCP) at Iquitos (Peru) has been breeding A.
nancymaae in captivity since 1979 and A. vociferans since 1984. Gozalo
and Montoya [65] showed that both species are not affected in the same
way by the same diseases. The main causes of death in A. nancymaae
were acute lobular pneumonia (24.2%), acute catarrhal enteritis (9.7%),
acute hemorrhagic enteritis (8.1%) and chronic nephropathy (8.1%),
whereas, for A. vociferans, they were acute lobular pneumonia (26.4%),
chronic nephropathy (13.2%), acute catarrhal enteritis (7.5%) and
acute toxic hepatitis (7.5%).
In addition, both species were different in their reproductive
parameters [65,66]. The mean age at first birth in females born in
captivity was 40.56 ± 7.82 months for A. nancymaae and 48.47 ± 12.67
months for A. vociferans. The youngest age of conception in captive
born females was 20.5 months in A. nancymaae and 31 months in
A. vociferans; the survival rate of live offspring at the first year of age
was 85.5% for A. nancymaae and 77.8% for A. vociferans. Birth peaks
occurred between October and January for A. nancymaae, and between
December and March for A. vociferans[66].
The two species further differ in their use of the forest canopy. In
the Tahuayo River, A. nancymaae had 21.4% of its sleeping sites in the
shrub stratum of the under story, 64.3% in the lower story and 14.3% in
the middle story. No sleeping sites were found in the upper story or in
emergent trees for this species. In the Nanay and Napo river forests, the
sleeping sites of A. vociferans followed a different distribution: 11.4%
in the under story, 54.3% in the lower story, 20% in the middle story,
8.6% in the upper story and 5.7% in emergent trees. In other words,
the sleeping sites for A. nancymaae were below a height of 25 m, while
those for A. vociferans were up to an elevation of 37 m, meaning it
utilizes higher sleeping sites [67].
Finally, the population densities of both species were different in the
Peruvian Amazon. Densities of A. nancymaae were 46.3 individuals/
km2 in lowland forests and 24.2 individuals/km2 in highland forests,
whereas the A. vociferans densities were 33 individuals/km2 in lowland
forests and 7.9 individuals/km2 in highland forests [67]. The lower
densities recorded for highlands forest could be related to the scarcity
of suitable nesting sites, particularly hollow tree trunks exclusively
employed by A. vociferans. The average group size was 4.1 individuals
for A. nancymaae and 3.3 individuals for A. vociferans in the same
study. Therefore, in both types of forests (but especially in highland
forests), A. nancymaae seems to have a demographic advantage.
Given these data, if FIDIC legally extracts considerable numbers of
A. vociferans (1,600 specimens, during eight months, in September 13,
2006 and then the contract was renewed in 2008) on the Colombian
side and later releases them in combination with exemplars of A.
nancymaae illegally obtained from Peru and Brazil, then A. nancymaae
via competition could clearly displace the natural Aotus form in the
Colombian Amazon, as it appears to have begun occurring. The
intermixing of specimens of these two Aotus taxa, in these releases,
may alter the evolutionary and genetic patterns that have naturally
occurred in the Amazon over millions of years. Therefore, it is urgent
that the environmental authorities of Colombia, Peru and Brazil (or
J Primatol
ISSN: 2167-6801 JPMT, an open access journal
Page 7 of 9
other international authorities) consider the explained situation and
halt the illegal trade of Aotus.
Some phylogenetic notes on Aotus
Although the main aim of this work is to show the use of molecular
markers to detect illegal traffic of Aotus in Colombia, some interesting
phylogenetic insights on this genus could be mentioned. This is the
fifth study which employs mtDNA sequences to understand the
phylogeny of Aotus. The first two used the same marker that we
employed here mtCOII [2,3]. Both studies only analyzed six and 12
individuals, respectively. The third study analyzed five mitochondrial
genes and one Y-linked gene but in only 18 individuals. The fourth
work also employed mtCOII, but the number of exemplars analyzed
was higher (69 individuals [4]. In the current work, we analyzed 111
individuals constituting the highest sample analyzed for this genus
from a molecular point of view until the present.
With this increase of animals analyzed from different taxa,
some phylogenetic patterns could be seen. We agree with Ford’s
perspective [1994] with regard to the northern Aotus. This author
only distinguished two northern species of Aotus, A. vociferans and
A. trivirgatus. Our analysis showed that there was not reciprocal
monophyly among the haplotypes of A. vociferans, A. brumbacki, A.
lemurinus, A. jorgehernandezi and A. griseimembra, although some
clades were composed exclusively by haplotypes of A. vociferans or A.
griseimembra. This could mean that all these Aotus taxa have recently
diverged and genetic divergence is very low, although they have
different karyotypes. The phylogenetic relationships among Aotus are
complicated because stasipatric speciation [68,69] is common within
this genus. This speciation process predicts the fixation of new mutant
homokaryotypes and the generation of reproductive barriers in only a
few generations (theoretically two or three) if the effective population
size is extremely small with endogamous mating, as is the case in Aotus
[68]. This could produce very different karyotypes in neighbouring
populations of Aotus, which are practically undifferentiated from
molecular sequences because they are strongly related from a
phylogenetic perspective. In fact, one or few families of Aotus could
suffer stasipatric processes and in a few generations could generate
individuals with different incompatible karyotypes with the remainder
individuals of its own population without any geographical barrier.
For this reason, we agree with the picture of Ford (1994) and the taxa
named A. vociferans, A. brumbacki, A. lemurinus, A. jorgehernandezi
and A. griseimembra should be treated as a superspecies in the same
way that the rodent Spalax ehrenbergii from the Middle East has been,
with animals of 2n=48, 52, 54, 56 and 58 chromosomes all being
included in this taxon [70,71]. This super species should be named
A. vociferans because it is the oldest name (1823) (brumbacki-1983;
lemurinus-1843; jorgehernandezi-2007; griseimembra-1912). Whether
A. zonalis belongs to this superspecies or whether A. trivirgatus is a
fully differentiated species with a more elevated genetic divergence
from A. vociferans must be investigated.
As it was also claimed by Ford (1994), A. nancymaae and A.
miconax belong to the same clade and the two A. miconax haplotypes
were separately intermixed with the haplotypes of A. nancymaae. This
group should be named A. miconax because this name is older than
A. nancymaee (1927 vs. 1983; A. miconax miconax and A. miconax
nancymaee).
We again agree with Ford (1994) regarding A. azarae azarae and
A. azarae boliviensis. They are two well-differentiated taxa. We did not
Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107
enclose samples from A. infulatus, but, if future works could show a
strong molecular relationship [72-79] between infulatus and boliviensis,
we could speak of A. azarae and A. infulatus boliviensis and A. infulatus
infulatus. If there was not a striking molecular relationship between
infulatus and boliviensis, then we could name the Bolivian animals as
A. Boliviensis.
Acknowledgments
We would like to acknowledge the SDA (Secretaria Distrital Ambiental
of Bogota DC, Colombia) for the project entitled “Fortalecimiento del control
y prevención del tráfico ilegal de fauna silvestre, especialmente de Primates,
a través de la determinación de zonas sometidas a extracción ilegal utilizando
pruebas de genética molecular de poblaciones”, and to Corpoamazonía (Leticia-
Colombia), which allowed us to obtain the necessary financial resources to carry
out the current study. Additional thanks to P. Escobar-Armel for their respective
help in obtaining Aotus samples during the last seven years. Many thanks goes
to the Bolivian and Peruvian Ministry of Environment, to the Dirección General
de Biodiversidad, to the Wildlife Conservation Society and CITES from Bolivia, to
the PRODUCE, Dirección Nacional de Extracción and Procesamiento Pesquero
from Peru, and the Consejo Nacional del Ambiente and the Instituto Nacional de
Recursos Naturales (INRENA) for their role in facilitating the obtainment of the
collection permits. We would also like to acknowledge the Colección Boliviana de
Fauna (Dr. Julieta Vargas) in La Paz (Bolivia). We also thank the Ticuna, Yucuna,
Yaguas, Witoto and Cocama Indian communities in the Colombian Amazon, Bora,
Ocaina, Shipibo-Comibo, Capanahua, Angoteros, Orejón, Cocama, Kishuarana
and Alama in the Peruvian Amazon, and the Movima, Moxeño, Sirionó, Canichana,
Cayubaba and Chacobo Indian communities in Bolivia.
References
1. Hershkovitz P (1983) Two new species of night monkeys, genus Aotus
(Cebidae, Platyrrhini): A preliminary report on Aotus taxonomy. Am J Primatol
4: 209-243.
2. Ashley MV, Vaughn JL (1995) Owl monkeys (Aotus) are highly divergent in
mitochondrial cytochrome c oxidase (COII) sequences. Int J Primatol 16: 793-
806.
3. Plautz HL, Gonçalves EC, Ferrari SF, Schneider MP, Silva A (2009)
Evolutionary inferences on the diversity of the genus Aotus (Platyrrhini,
Cebidae) from mitochondrial cytochrome c oxidase subunit II gene sequences.
Mol Phylogenet Evol 51: 382-387.
4. Ruiz-García M, Vásquez C, Camargo E, Leguizamon N, Gálvez H, et al. (2011)
Molecular phylogenetics of Aotus (Platyrrhini, Cebidae). Int J Primatol 32:
1218-1241.
5. Schneider H, Schneider MPC, Sampaio I, Harada ML, Stanhope M, et al.
(1993) Molecular phylogeny of the New World monkeys (Platyrrhini, Primates).
MOL PHYLOGENET EVOL 2: 225-242.
6. Schneider H, Sampaio I, Harada ML, Barroso CM, Schneider MP, et al. (1996)
Molecular phylogeny of the New World monkeys (Platyrrhini, primates) based
on two unlinked nuclear genes: IRBP intron 1 and epsilon-globin sequences.
Am J Phys Anthropol 100: 153-179.
7. Schneider H, Canavez FC, Sampaio I, Moreira MA, Tagliaro CH, et al. (2001)
Can molecular data place each neotropical monkey in its own branch?
Chromosoma 109: 515-523.
8. Harada ML, Schneider H, Schneider MP, Sampaio I, Czelusniak J, et al.
(1995) DNA evidence on the phylogenetic systematics of New World monkeys:
Support for the sister-grouping of Cebus and Saimiri from two unlinked nuclear
genes. Mol Phylogenet Evol 4: 331-349.
9. Opazo JC, Wildman DE, Prychitko T, Johnson RM, Goodman M (2006)
Phylogenetic relationships and divergence times among New World monkeys
(Platyrrhini, Primates). Mol Phylogenet Evol 40: 274-280.
10. Schmidt LH (1973) Infections with Plasmodium falciparum and Plasmodium vivax
in the owl monkey-model systems for basic biological and chemotherapeutic
studies. Trans R Soc Trop Med Hyg 67: 446-474.
11. Schmidt LH (1978) Plasmodium falciparum and Plasmodium vivax infections in
the owl monkey (Aotus trivirgatus). I. The courses of untreated infections. Am J
Trop Med Hyg 27: 671-702.
12. Collins WE, Stanfill PS, Skinner JC, Harrison AJ, Smith CS (1974) Studies on
human malaria in Aotus monkeys. IV. Development of Plasmodium falciparum
in two subspecies of Aotus trivirgatus. J Parasitol 60: 355-358.
J Primatol
ISSN: 2167-6801 JPMT, an open access journal
Page 8 of 9
13. Taylor DW, Siddiqui WA (1979) Susceptibility of owl monkeys to Plasmodium
falciparum infection in relation to location of origin, phenotype, and karyotype.
J Parasitol 65: 267-271.
14. Brumback RA, Willenborg DO (1973) Serotaxonomy of Aotus. A preliminary
study. Folia Primatol 83: 100-125.
15. Gozalo A, Ballou WR, Lucas CM, Wellde B, Hall BT, et al. (1996) Efficacy of
passively transferred rabbit antibody to two different recombinant Plasmodium
falciparum MSP-1 constructs in infected Aotus nancymai. American Society of
Tropical Medicine and Hygiene, Baltimore, Maryland.
16. Spirig R, Peduzzi E, Patarroyo ME, Pluschke G, Daubenberger CA (2005)
Structural and functional characterisation of the Toll like receptor 9 of Aotus
nancymaae, a non-human primate model for malaria vaccine development.
Immunogenetics 57: 283-288.
17. Baquero JE, Miranda S, Murillo O, Mateus H, Trujillo E, et al. (2006) Reference
strand conformational analysis (RSCA) is a valuable tool in identifying MHC-
DRB sequences in three species of Aotus monkeys. Immunogenetics 58: 590-
597.
18. Patarroyo ME, Cifuentes G, Baquero J (2006) Comparative molecular and
three-dimensional analysis of the peptide-MHC II binding region in both human
and Aotus MHC-DRB molecules confirms their usefulness in antimalarial
vaccine development. Immunogenetics 58: 598-606.
19. Suárez CF, Patarroyo ME, Trujillo E, Estupiñán M, Baquero JE, et al. (2006)
Owl monkey MHC-DRB exon 2 reveals high similarity with several HLA-DRB
lineages. Immunogenetics 58: 542-558.
20. Daubenberger CA, Spirig R, Patarroyo ME, Pluschke G (2007) Flow cytometric
analysis on cross-reactivity of human-specific CD monoclonal antibodies with
splenocytes of Aotus nancymaae, a non-human primate model for biomedical
research. Vet Immunol Immunopathol 119: 14-20.
21. Defler TR (2003) Primates de Colombia. Conservación Internacional Colombia,
Bogotá.
22. Defler TR, Bueno ML (2007) Aotus diversity and the species problem. Primate
Conservation 22: 55-70.
23. Ruiz-García M, Alvarez D (2003) RFLP analysis of mtDNA from six platyrrhine
genera: phylogenetic inferences. Folia Primatol (Basel) 74: 59-70.
24. Ruiz-García M (2005) The use of several microsatellite loci applied to 13
Neotropical Primates revealed a strong recent bottleneck event in the woolly
monkey (Lagothrix lagotricha) in Colombia. Primate Rep 71: 27-55.
25. Ruiz-García M, Álvarez D, Nassar F, Avila A, Rojas S,et al. (1999) Preliminary
RFLPs mtDNA analysis in five Colombian Primate species (Cebus apella,
Cebus albifrons, Saimiri sciureus, Lagothrix lagotricha and Alouatta seniculus):
Mitochondrial Genetic diversity analysis. Primates 40: 78-94.
26. Ruiz-Garcia M (2003) Molecular population genetic analysis of the spectacled
bear (Tremarctos ornatus) in the northern Andean area. Hereditas 138: 81-93.
27. Ruiz-García M, Parra A, Romero-Aleán N, Escobar-Armel P, Shostell JM
(2006) Genetic Characterization and Phylogenetic Relationships between the
Ateles Species (Atelidae, Primates) by Means of DNA Microsatellite Markers
and Craniometric Data. Primate Rep 73: 3-48.
28. Ruiz-Garcia M, Escobar-Armel P, Alvarez D, Mudry M, Ascunce M, et al. (2007)
Genetic variability in four Alouatta species measured by means of nine DNA
microsatellite markers: genetic structure and recent bottlenecks. Folia Primatol
(Basel) 78: 73-87.
29. Ruiz-Garcia M, Pinedo-Castro MO (2010) Molecular systematics and
phylogeography of the genus Lagothrix (Atelidae, Primates) by means of the
mitochondrial COII gene. Folia Primatol (Basel) 81: 109-128.
30. Ruiz-Garcia M, Castillo MI, Ledezma A, Leguizamon N, Sánchez R, et al.
(2012) Molecular systematics and phylogeography of Cebus capucinus
(Cebidae, Primates) in Colombia and Costa Rica by means of the mitochondrial
COII gene. Am J Primatol 74: 366-380.
31. Ruiz-García M, Castillo MI, Lichilín N, Pinedo-Castro M (2012b) Taxonomy
considerations and molecular phylogeography of Cebus apella and related taxa
with the mtCOII gene. Folia Primatologica (in press).
32. Ruiz-García M, Pacheco L, Alvarez D (2009) Genetic characterization of the
Bolivian Andean puma (Puma concolor) at the Sajama National Park (SNP)
and relationships with other north-western South American puma populations.
Rev. chil. hist. nat. 82: 97-117.
Volume 2 • Issue 1 • 1000107
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF, Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII
Gene Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini, Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-
6801.1000107
33. Ruiz-García M, Martínez-Aguero M, Alvarez D, Goodman S (2009)
Genetic variability in Neotropical deer genera (Mammalia: Cervidae) as loci
microsatellite. Rev. Biol. San Jose trop 57: 879-904.
34. Ruiz-García M, Castillo MI, Vásquez C, Rodriguez K, Pinedo-Castro M, et
al. (2010) Molecular phylogenetics and phylogeography of the white-fronted
capuchin (Cebus albifrons; Cebidae, Primates) by means of mtCOII gene
sequences. Mol Phylogenet Evol 57: 1049-1061.
35. Ruiz-García M, Orozco-terWengel P, Castellanos A, Arias L (2005)
Microsatellite analysis of the spectacled bear (Tremarctos ornatus) across its
range distribution. Genes Genet Syst 80: 57-69.
36. Ruiz-García M, Shostell J (2010) Biology, Evolution, and Conservation of River
Dolphins within South America and Asia. Nova Science Publishers Inc, New
York.
37. Ruiz-Garcia M, Payán E, Murillo A, Alvarez D (2006) DNA microsatellite
characterization of the jaguar (Panthera onca) in Colombia. Genes Genet Syst
81: 115-127.
38. Ruiz-García M, Murillo A, Corrales C, Romero-Aleán N, Alvarez-Prada D
(2007) Amazonian population genetics: the evolutionary history of the jaguar,
ocelot, pink dolphin, and wolly monkey and wattled curassow reconstructed
from their genes. Anim Biodivers Conserv 30: 115-130.
39. Capaldi RA (1990) Structure and function of cytochrome c oxidase. Annu Rev
Biochem 59: 569-596.
40. Adkins RM, Honeycutt RL (1994) Evolution of the primate cytochrome c
oxidase subunit II gene. J Mol Evol 38: 215-231.
41. Adkins RM, Honeycutt RL (1991) Molecular phylogeny of the superorder
Archonta. Proc Natl Acad Sci U S A 88: 10317-10321.
42. Ruvolo M, Disotell TR, Allard MW, Brown WM, Honeycutt RL (1991) Resolution
of the African hominoid trichotomy by use of a mitochondrial gene sequence.
Proc Natl Acad Sci U S A 88: 1570-1574.
43. Disotell TR, Honeycutt RL, Ruvolo M (1992) Mitochondrial DNA phylogeny of
the Old-World monkey tribe Papionini. Mol Biol Evol 9: 1-13.
44. Figueiredo WB, Carvalho-Filho NM, Schneider H, Sampaio I (1998)
Mitochondrial DNA sequences and the taxonomic status of Alouatta seniculus
populations in Northeastern Amazonia. Neotrop Primates 6: 73-77.
45. Collins AC, Dubach JM (2000) Phylogenetic relationships of spider monkeys
(Ateles) based on mitochondrial DNA variation. Int J Primatol 21: 381-420.
46. Collins AC, Dubach JM (2000b) Biogeographic and ecological forces
responsible for speciation in Ateles. Int J Primatol 21: 421-444.
47. Nieves M, Ascunce MS, Rahn MI, Mudry MD (2005) Phylogenetic relationships
among some Ateles species: the use of chromosomic and molecular characters.
Primates 46: 155-164.
48. Ascunce MS, Hasson E, Mudry MD (2003) COII: a useful tool for inferring
phylogenetic relationships among New World monkeys (Primates, Platyrrhini).
Zool Scripta 32: 397-406.
49. Sena L, Vallinoto M, Sampaio I, Schneider H, Ferrari SF, et al. (2002)
Mitochondrial COII gene sequences provide new insights into the phylogeny
of marmoset species groups (Callitrichidae, Primates). Folia Primatol 83: 100-
125.
50. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory
manual. (2nd edn) Cold Spring Harbor Laboratory Press, New York.
51. Walsh PS, Metzger DA, Higuchi R (1991) Chelex 100 as a medium for simple
extraction of DNA for PCR-based typing from forensic material. Biotechniques
10: 506-513.
52. Chung WK, Steiper ME (2008) Mitochondrial COII introgression into the nuclear
genome of Gorilla gorilla. Int J Primatol 29: 1341-1353.
53. Saitou N, Nei M (1987) The neighbor-joining method: a new method for
reconstructing phylogenetic trees. Mol Biol Evol 4: 406-425.
54. Reis NR, Peracchi AL, Andrade FR (2008) Primatas Brasileiros. Technical
Books Editora, Brazil.
55. Nassar F, Pereira V, Savage A (2003) Primatology New World Biología,
Medicina, Manejo y Conservación. Araguatos Primatology Center,
Bogota,Colombia.
56. Nassar F, Pereira V, Savage A (2003a) Primatología del Nuevo Mundo:
J Primatol
ISSN: 2167-6801 JPMT, an open access journal
Page 9 of 9
Biología, Medicina, Manejo, Conservación. Centro de Primatología Araguatos.
Bogota,Colombia.
57. Koven VT (2008) Population Genetics Research Progress. Nova Publisher
Inc.,New York.
58. Aquino R, Álvarez J, Mulanovich A (2005) Diversity and conservation status
of primates in the Sierras of contaminates, Peruvian Amazon. Rev Peru Biol
12: 427-434.
59. Lopes MA, Rehg JA (2003) Observations of Callimico goeldii with Saguinus
imperator in the serra do Divisor National Park, Acre, Brazil. Neotrop Primates
11: 181-183.
60. Puertas P, Bodmer RE (1993) Conservation of a high diversity Primate
assemblage. Biodiversity and Conservation 2: 586-593.
61. Aquino R, Encarnación E (1988) Population densities and geographic
distribution of night monkeys (Aotus nancymai and Aotus vociferans) (Cebidae:
Primates) in northeastern Peru. Am J Primatol 14: 375-381.
62. Auricchio P (1995) Primates in Brazil. Terra Brasilis.
63. Torres OM, Enciso S, Ruiz F, Silva E, Yunis I (1998) Chromosome diversity of
the genus Aotus from Colombia. Am J Primatol 44: 255-275.
64. Ma NS, Jones TC, Miller AC, Morgan LM, Adams EA (1976) Chromosome
polymorphism and banding patterns in the owl monkey (Aotus). Lab Anim Sci
26: 1022-1036.
65. Gozalo A, Montoya E (1990) Mortality causes of owl monkeys (Aotus nancymae
and Aotus vociferans) in captivity. J Med Primatol 19: 69-72.
66. Gozalo A, Montoya E (1990b) Reproduction of the owl monkey (Aotus
nancymai) (Primates: Cebidae) in captivity. Am J Primatol 21: 61-68.
67. Aquino R, Encarnación F (1986) Characteristics and use of sleeping sites in
Aotus (Cebidae: Primates) in the Amazon Lowlands of Peru. Am J Primatol
11: 319-331.
68. White MJ (1968) Models of speciation. New concepts suggest that the classical
sympatric and allopatric models are not the only alternatives. Science 159:
1065-1070.
69. White MJD (1978) Modes of speciation. W. H. Freeman, San Francisco.
70. Nevo E (1991) Evolutionary Theory and processes of active speciation
and adaptative radiation in subterranean mole rats, Spalax ehrenbergi
superspecies, in Israel. Evolutionary Biology 25: 1-125.
71. Montoya E, Moro J, Gozalo A, Samamé H (1994) Reproduction of Aotus
vociferans (primates CEBIDAE) in captivity. livestock Research Journal IVITA.
72. Ho SY, Saarma U, Barnett R, Haile J, Shapiro B (2008) The effect of
inappropriate calibration: three case studies in molecular ecology. PLoS One
3: e1615.
73. Atchley WR, Woodruff DS (1981) Evolution and speciation. Cambridge
University Press, Cambridge.
74. Rodríguez NC (1990) Primatology in Peru: primatological research (1973-
1985) Proyecto Peruano de Primatología, Lima, Peru.
75. Baer JF, Seller RE, Kakoma I (1994) Aotus: The Owl Monkey. Elsevier Science
and Tech.
76. Masters JC, Boniotto M, Crovella S, Roos C, Pozzi L, et al. (2007) Phylogenetic
relationships among the Lorisoidea as indicated by craniodental morphology
and mitochondrial sequence data. Am J Primatol 69: 6-15.
77. Harada-Hamel ML, Schneider H, Encarnación F, Montoya E, Villavicencio E
(1990) ABO groups in two species of Peruvian night monkeys (Aotus nancymai
and Aotus vociferans). Rev Bras Genet.
78. Thorington RW, Heltne PG (1976) Neotropical primates: field studies and
conservation: proceedings of a symposium on the distribution and abundance
of neotropical primates. National Academy of Sciences, Washington DC.
79. de Mello MT (1986) A Primatologia No Brasil. (2ndedn), Sociedade Brasileira
de Primatologia, Brazil.
Citation: Ruiz-García M, Vásquez C, Camargo E, Castellanos-Mora LF,
Gálvez H, et al. (2013) Molecular Genetics Analysis of mtDNA COII Gene
Sequences Shows Illegal Traffic of Night Monkeys (Aotus, Platyrrhini,
Primates) in Colombia. J Primatol 2:107. doi:10.4172/2167-6801.1000107
Volume 2 • Issue 1 • 1000107